Isolation of recombinant antibodies against EspA and intimin of Escherichia coli O157:H7

Intimin, Tir, and EspA proteins are expressed by attaching-effacing Escherichia coli, which include enteropathogenic and enterohemorrhagic E. coli pathotypes. EspA proteins are part of the type three secretion system needle complex that delivers Tir to the host epithelial cell, while surface arrayed intimin docks the bacterium to the translocated Tir. This intimate attachment leads to attaching and effacing lesions. Recombinant forms of these effector proteins from enterohemorrhagic E. coli O157:H7 were produced by using E. coli expression vectors. Binding of intimin and Tir fragments in enzyme-linked immunosorbent assay (ELISAs) demonstrated the interaction of intimin fragments containing the C-terminal 282 or 188 amino acids to a Tir fragment containing amino acid residues 258 to 361. Recombinant intimin and EspA proteins were used to elicit immune responses in rabbits and immune phage-display antibody libraries were produced. Screening of these immune libraries by conventional phage-antibody panning and colony filter screening produced a panel of antibodies with specificity for EspA or intimin. Antibodies recognizing different C-terminal epitopes on intimin bound specifically to the gamma intimin of O157:H7 and not to other classes of intimin. Antibodies recognizing EspA from E. coli O157 also recognized the protein from the eae-deficient O157 mutant DM3 and from E. coli O111. Anti-intimin antibodies were also produced as fusion proteins coupled to the reporter molecule alkaline phosphatase, allowing the one-step detection of gamma intimin. The isolated recombinant monoclonal antibodies were functional in a range of assay formats, including ELISA, Western blotting, and dot blots, thus demonstrating their diagnostic potential.

Identification and phylogenetic inferences on stocks of sharks affected by the fishing industry off the Northern coast of Brazil

The ongoing decline in abundance and diversity of shark stocks, primarily due to uncontrolled fishery exploitation, is a worldwide problem. An additional problem for the development of conservation and management programmes is the identification of species diversity within a given area, given the morphological similarities among shark species, and the typical disembarkation of processed carcasses which are almost impossible to differentiate. The main aim of the present study was to identify those shark species being exploited off northern Brazil, by using the 12S-16S molecular marker. For this, DNA sequences were obtained from 122 specimens collected on the docks and the fish market in Bragança, in the Brazilian state of Pará. We identified at least 11 species. Three-quarters of the specimens collected were either Carcharhinus porosus or Rhizoprionodon sp, while a notable absence was the daggernose shark, Isogomphodon oxyrhyncus, previously one of the most common species in local catches. The study emphasises the value of molecular techniques for the identification of cryptic shark species, and the potential of the 12S-16S marker as a tool for phylogenetic inferences in a study of elasmobranchs.

Políticas e tensões entre o porto e o espaço escolar em vila do conde – Barcarena/PA

O presente texto apresentará um estudo realizado na escola Wandick Gutierrez em Vila do Conde, no município de Barcarena no estado do Pará. O objetivo é compreender de que forma se materializa as práticas de educação ambiental no contexto escolar a partir da inserção do Programa Saberes e Práticas de Responsabilidade Social na escola Wandick Gutierrez: ações socioeducativas nos bairros do entorno do Porto de Vila do Conde – PROSPRS realizado em parceria pela Universidade Federal do Pará- UFPA, através do Grupo de Pesquisa em Educação, Cultura e Meio ambiente- GEAM e a Companhia Docas do Pará – CDP, no período de 2011 a 2012. O referencial teórico metodológico amparou-se no materialismo histórico dialético em uma perspectiva da educação ambiental crítica; esta contou com a análise de documentos relacionados ao programa, bem como documentos da escola e, revisão de literatura e vivências no cotidiano da escola pesquisada. A pesquisa contou com entrevistas semiestruturadas direcionadas a direção da escola, professores e coordenação pedagógica; aplicação de questionários, junto a alunos, responsáveis de alunos, equipe do programa e a CDP. Alguns dos autores que nos auxiliaram para a compreensão do contexto estudado foram: Marx (1967, 1998) Frederico Loureiro (2005, 2007), Marilena Loureiro (2007), Saviani (2006), Paulo Freire (1996, 2000), Frigotto (2012) Oliveira (2005), dentre outros. A pesquisa oportunizou perceber que nos discursos dos sujeitos entrevistados e, na leitura dos relatórios do PROSPRS houve tentativas de relacionar as ações do programa a partir da realidade concreta da comunidade da Vila do Conde; as ações desenvolvidas pelo programa acabaram por desencadear vivências/percepções ambientais nos sujeitos escolares ainda não vividas por estes, a exemplo a construção de hortas e compostagem no ambiente escolar, no entanto, compreendo que tais discursos ainda necessitam de conhecimentos mais aprofundados do próprio fazer ambiental. Contudo, as ações ambientais que se iniciaram no ambiente escolar Wandick Gutierrez são relevantes para uma discussão reflexiva crítica para todos os sujeitos envolvidos nesse processo.

Problemas ambientais e relações de trabalho no Porto de Belém: perspectivas para um projeto de educação ambiental

Esta pesquisa mostra o olhar que trabalhadores e passageiros da Companhia das Docas do Pará – Porto de Belém têm do cotidiano do porto em relação aos problemas ambientais. Nossa motivação para o desenvolvimento deste estudo pautou-se na identificação de qual a situação ambiental na área portuária das Docas de Belém e se as ações de Educação Ambiental desenvolvidas em seu interior contribuem para a construção de melhorias socioambientais? Para a realização da pesquisa utilizamos como forma de análise a abordagem qualitativa, na qual buscamos priorizar não apenas a quantidade de dados referentes ao objeto de estudo, mas principalmente compreender o fenômeno em foco. No estudo desenvolvemos as seguintes etapas: a) levantamos dados sobre o entorno, destacando a ocupação urbana na área de influência do Porto; b) elaboramos um diagnóstico ambiental, por intermédio da visão de trabalhadores e usuários do Porto de Belém, visando suas expectativas quanto ao ambiente; c) Buscamos compreender o desenvolvimento de ações relacionadas à preservação ambiental, necessárias ao processo de educação para a sustentabilidade, desenvolvidas no Porto de Belém. Neste sentido, inicialmente realizamos uma ampla revisão da literatura sobre a temática, conjuntamente com a pesquisa documental, onde analisamos textos escritos por pesquisadores na área de meio ambiente, e da educação Ambiental, como: Loureiro (2004, 2009), Lima (2007), Silva (2009), Teisserenc (2010), Tapia (2005), Campanhola (2000), Gadotti (2008), Becker (2008), Antunes (2002). Assim como documentos oficiais relacionados ao meio ambiente e a outras iniciativas voltadas a Ações de Preservação e de Portos como: Constituição da Republica Federativa (1988); Decreto Lei 9.795 que dispõe sobre a educação ambiental, institui a Política Nacional de Educação Ambiental; Decreto Lei 8.630 que dispõe sobre o regime jurídico da exploração dos portos organizados e das instalações portuárias; Lei nº 8.630, de 25/02/1993 (antiga Lei dos Portos). A partir da construção do texto ampliamos nosso conhecimento, o que nos deu suporte teórico para iniciarmos a pesquisa empírica. Nesta etapa fizemos um levantamento da área edificada em parte da orla de Belém para identificar o percentual de ocupação urbana na faixa exclusiva selecionada, às proximidades do lócus da pesquisa, o levantamento realizado visou identificar alguma provável influência externa na área de movimentação de navios, o que foi descartado, pois o percentual de ocupação urbana (45,0830 %) encontrado foi considerado dentro dos padrões aceitáveis. Na segunda etapa do estudo foi feita a coleta de dados junto às pessoas que vivenciam o cotidiano da área portuária como: trabalhadores e usuários, objetivando obter informações sobre sua visão e opinião sobre o foco da pesquisa, utilizamos primeiramente a entrevista semiestruturada, objetivando esclarecer sobre as reais condições atuais do meio ambiente de trabalho, vivido cotidianamente por seus atores, nas questões profissionais, sociais e ambientais na área do Porto de Belém e, para que tivéssemos um diagnóstico do momento em que se encontra a ambientalização do Porto, realizamos entrevistas com sujeitos que trabalham diariamente naquele ambiente portuário. Culminando o estudo, utilizamos questionários, os quais foram aplicados a 80 (oitenta) sujeitos trabalhadores e 149 (cento e quarenta e nove) aos usuários/passageiros. O resultado da pesquisa revelou como principais problemas apontados, tanto pelos usuários como pelos trabalhadores os impactos ambientais causados pelas atividades portuárias como a emissão de resíduos sólidos, líquidos, derrame de produtos perigosos e os riscos de contaminação do rio. Assim como lixo em excesso e sua coleta indevida, que tem ocasionado frequentes entupimentos dos esgotos. Assim como a inexistência de continuidade do trabalho de limpeza e manutenção com a prática necessária da dragagem retirada de entulhos do rio e do canal das docas. Constatamos também, que existe a intenção pela gestão da CDP em realizar o trabalho de utilização correta de sistemas de carga e descarga, armazenagem, pesagem, acomodação e movimentação de cargas no porto, bem como realizar de forma correta controle de matérias que chegam e que saem, e que é perceptível a eficiente vigilância da movimentação na área de embarque e desembarque de passageiros, o fazem com que o Porto de Belém permaneça dentro dos padrões estabelecidos na Constituição, o processo de gestão portuária ocorre regularmente obedecendo a um sistema de atividades previamente determinadas e que se encontram de acordo com a Lei dos Portos. O grande ganho que se observa no estudo e esclarecimento das questões investigadas é a apresentação do conceito que os atores envolvidos têm da realidade ambiental existente.

Hidrocarbonetos policíclicos aromáticos em sedimentos e organismos bentônicos do Terminal de Miramar (Baía do Guajará - Belém - Pará - Amazônia)

ABSTRACT: In Guajará bay, mouth of the Amazon River, is located the capital of Pará state (Belém) and its metropolitan area. Exists in this area an intense boat traffic, as well as transportation and sale of fuels in floating docks and activities related to the storage and transportation of oil in Miramar Petrochemical Terminal (TEMIR). Small spills and discharges of oil in water can serve as sources of pollution by polycyclic aromatic hydrocarbons (PAHs). PAHs are organic compounds generated by incomplete combustion of organic matter (OM) and are among the contaminants of most interest in environmental studies due to their mutagenic and carcinogenic potential. One way to detect and evaluate the impact of PAHs in an environment is using biomonitors, however the qualitative and quantitative analysis in sediments are most widely used. This study aimed to evaluate, by Gas Chromatography/Mass Spectrometry (GC/MS), the 16 PAHs considered as priority by the Environmental Protection Agency of the United States in sediments and benthic organisms (Namalicastys abiuma) of TEMIR. Field expeditions were carried out in December 2012, March, May and June 2013 representing the dried, wet (two samples) and dried seasons successively. Fine sediments levels dominated in both sampling periods and the %Fines in Guajará bays has a direct relationship to the %OM. In relation to the 16 PAHs studied, 10 of them were detected in the wet season sediments samples and 8 in the dried season. Even with a low diversity of aromatic compounds, sediment samples of the dried season presented greater ΣHPAs (1.351,43 ng g^-1) than the sediments of the wet season (263,99 ng g^-1), which can be related to the increased hydrodynamic in Guajará bay this last period. Correlation analysis indicated that ΣHPAs not seem to be influenced by %Fine and %OM. Benzo(a)pyrene accounted for 87% of ΣHPAs during the wet season, the other PAHs showed percentage ≤ 3%. During the other period stood out: pyrene (18% of ΣHPAs) fluoranthene (16%), the chrysene, benzo(b)fluoranthene (15%) and benzo(a)pyrene (11%). The use of the geochemical reason to interpret likely sources of PAHs indicated that there is a predominance of aromatics from pyrolytic origin in TEMIR. The activities that may be responsible for the PAHs levels observed in sediments and benthic organisms in TEMIR are the discharge of untreated domestic sewage and the emissions of particulates from cars and small/medium boats. The ΣHPAs seems to influence the density of polychaetes because a reduction of about 50% in the number of organisms was observed during the dry season when there was the greater amount of ΣHPAs in their body structure. Among the 16 PAHs studied, 11 were detected in the polychaetes during the dry season (ΣHPAs_MED = 848,71 ng g^-1) and 10 in the wet season (ΣHPAs_MED = 141,85 ng g^-1). Were highlighted during the dried season: indeno(1,2,3-c,d)pyrene (47%) and pyrene (23%). While in the wet season: pyrene (23%), chrysene (17%), fluorene (17%) and fluoranthene (13%). The %Rec obtained was >>100% indicating a matrix effect and reducing the quantitative accuracy of results. The use of the polychaete N. abiuma as biomonitor of pollution by PAHs in estuaries not seem to be effective when the sample period is limited, because collecting it requires a great effort to obtain a small amount of mass and it still results in a matrix effect in the chromatographic analysis (method %Rec >> 100%) that may not be corrected due to the restricted availability of sample mass. Studies with a larger number of samples, enabling depth statistical analysis, and covering other collection points in Guajará bay are required to prove mathematically that has been stated in this dissertation.

L' esprit des esprits, ou pensées choisies : pour servir de suite aux Maximes de la Rochefoucault

Vorlageform des Erscheinungsvermerks: A Londres, & se trouve à Hambourg chez Virchaux, Imprimeur-libraire

Estudio en modelo físico a escala reducida de la viabilidad del empleo de buques fondeados como diques flotantes para abrigo portuario y defensa de costas

Los diques flotantes son estructuras que atenúan la energía del oleaje fundamentalmente por reflexión y turbulencia. Aunque presentan importantes ventajas en términos constructivos y medioambientales, su efectividad es limitada y en la práctica sólo se emplean en condiciones climáticas propias de zonas con oleajes poco energéticos. Por otro lado, el buque es la estructura flotante por excelencia y su empleo para el abrigo portuario y costero en determinadas situaciones puede aportar las ventajas propias de los diques flotantes, al tiempo que ampliar el rango de oleajes frente a los que estas estructuras son efectivas. El propósito de esta Tesis Doctoral es evaluar la viabilidad del empleo de buques fondeados como diques flotantes para el abrigo portuario y costero. Para ello, se han realizado ensayos en modelo físico a escala reducida en un canal de oleaje del Centro de Estudios de Puertos y Costas (CEPYC), con el objeto de determinar los coeficientes de transmisión (Ct), reflexión (Cr) y disipación (Cd) de barcos de diversas tipologías y dimensiones, sometidos a diferentes oleajes en distintas situaciones de carga, fondeo y profundidad del emplazamiento. La efectividad de los buques empleados en los ensayos se ha determinado mediante el análisis de dichos coeficientes y su variación con la altura de ola y el periodo de los oleajes incidentes. Además, se han registrado las fuerzas existentes en las cadenas de fondeo con objeto de comprobar la viabilidad del mismo y facilitar una estimación del diámetro de las cadenas que serían necesarias en cada situación. Posteriormente, se han aplicado los resultados obtenidos en los ensayos en modelo físico reducido a dos situaciones de abrigo portuario y costero. La primera aplicación consiste en el empleo de buques como defensa temporal en fases constructivas por medios marítimos, partiendo de la hipótesis de que, actuando como diques flotantes, puede proteger la zona de la obra y ampliar las ventanas temporales de periodos de actividad en obra marítima. Las actividades que se han analizado son las de dragado de fondos, vertidos de material granular y transporte y fondeo de cajones flotantes para diques y muelles. La segunda aplicación estudiada es el empleo de buques para la protección costera y la formación de salientes y tómbolos. Los coeficientes de transmisión obtenidos se han introducido en formulaciones analíticas que permiten prever la evolución de la costa frente a la protección procurada por el buque actuando como dique flotante exento. Finalmente se han redactado las conclusiones de la investigación y se han propuesto nuevas líneas de investigación relacionadas con esta Tesis Doctoral. Floating breakwaters are structures which attenuate wave energy mainly by reflection and turbulence. They display advantages in terms of construction and ecology, amongst others. However, their use is restricted in practice to certain areas with good climatic conditions and low energy waves. Moreover, ships are the most common floating structures and their use for port and coastal shelter in certain situations could widen the range of applicability in addition to the rest of advantages of floating breakwaters. The purpose of this research is to assess the feasibility of ships anchored as floating breakwaters for port and coastal protection. To that end, tests in a scaled down physical model have been conducted in a wave flume in the Centre of Port and Coastal Studies (CEPYC), in order to determine the transmission (Ct), reflection (Cr) and dissipation (Cd) coefficients of ships of diverse types and dimensions, under different wave, load, anchoring and depth conditions. The effectiveness of the several ships used in the tests has been determined by analyzing these coefficients and their variation with the wave height and period of the incident waves. In addition, the existing forces in the anchor chains have been registered to verify the feasibility of the anchoring systems, as well as to provide an estimation of the diameter of the chains that would be needed in each situation. Subsequently, the results of the tests have been applied to two situations of port and coastal protection. The first one is the use of ships as a temporary defense for maritime works with construction phases by maritime means, on the assumption that, acting as floating breakwaters, they can protect the work area and increase the time windows of periods of activity in maritime works. Dredging, dumping of granular material and transport and positioning of big concrete caissons for docks and breakwaters were the activities analyzed. The second situation is the use of ships for coastal protection and forming salients of sand or tombolos. Some analytical formulations which take into account the transmission coefficients from the tests have been used to predict the evolution of the coastline under the protection given by the ships acting as detached floating breakwaters. Finally, the conclusions of the research have been addressed and the proposal of new lines of work related to the topic has been made.

Corrosión de armaduras en el hormigón armado en ambiente marino aéreo

La presente tesis doctoral surge ante la necesidad de completar el estudio sobre la corrosión en el hormigón armado expuesto al ambiente marino iniciado por D. Miguel Ángel Bermúdez Odriozola en 2007 con la tesis doctoral con título “Corrosión de las armaduras del hormigón armado en ambiente marino: zona de carrera de mareas y zona sumergida”. Una vez realizado el estudio bibliográfico, se ha planteado un estudio experimental en el que se analizan cuatro muelles de distintas edades y localizados en distintos lugares de la geografía española. Uno de ellos contiene adiciones en el hormigón. Una vez inspeccionados, se extraen testigos de cada uno de ellos con el fin de realizar ensayos de caracterización y se analizan sus resultados en paralelo con los coeficientes de difusión. Uno se los objetivos ha sido comprobar el mejor método de ensayo que permita controlar la durabilidad del hormigón en ambiente IIIa. La tesis que se presenta ha demostrado la efectividad del ensayo de penetración de agua para discriminar los hormigones que presentarán un buen comportamiento frente a la difusión de cloruros y por tanto para predecir la vida útil de la estructura. Asimismo, otro objetivo ha sido valorar el uso de adiciones en el hormigón y cómo afectan a la mejora del comportamiento del hormigón expuesto al ambiente marino. Se han obtenido valores del coeficiente de eficacia de cenizas volantes y humo de sílice frente a la difusión de cloruros, muy superiores a los actualmente considerados en términos de resistencia. El ambiente marino aéreo engloba estructuras situadas en primera línea de costa así como aquellas que se sitúan hasta 5km de distancia de la línea del mar. La investigación realizada ha demostrado la necesidad de establecer una nueva subdivisión en este ambiente introduciendo la zona Spray, siguiendo criterios internacionales, que permita diferenciar la zona más desfavorable del mismo que comprende las estructuras ubicadas sobre el nivel del mar tal como los tableros de pantalanes portuarios estudiados. Se han analizado en profundidad los valores del contenido de cloruros en superficie de hormigón en los diferentes tipos de ambiente marino recogidos por la Instrucción EHE- 08 contrastando los requisitos normativos. Como conclusión se han hecho nuevas propuestas normativas que afectan al ambiente sumergido y al ambiente marino aéreo (zona spray). Asimismo se ha realizado un análisis en profundidad de la concentración crítica de cloruros recogida por la Instrucción EHE-08, a partir de los datos obtenidos en estructuras reales. Se ha evaluado el ensayo de resistividad para estimar el valor del coeficiente de difusión de cloruros. Este ensayo ha demostrado su efectividad para discriminar hormigones con adiciones y con muy baja penetración de agua, si bien no resulta sensible para detectar hormigones con elevada permeabilidad. Finalmente en la tesis se ha obtenido un modelo de difusión de cloruros (a/c-D) en ambiente marino aéreo a fin de poder estimar la difusión de cloruros en el hormigón a partir de un requisito normativo como es la relación a/c. This Thesis is focussed to complete the previous study on the corrosion of reinforced concrete exposed to marine environment, carried out by Dr. Miguel Ángel Bermúdez Odrizola in 2007. That Thesis title was “Corrosion of concrete reinforcement located in marine environment: tidal zone and submerged zone”. After the literature review presented in the State of the Art, an experimental programme was carried out. Four docks of different ages and locations have been studied, one of them was a fly ash concrete. The structures were inspected on site and several cores extracted in order to develop characterization tests and obtain diffusion profiles at the laboratory. One of the objectives has been to check the best laboratory test method to control concrete durability when exposed to atmospheric marine environment. This Thesis has proven the effectiveness of the water penetration test to differentiate concretes well behaved against chloride diffusion and also to predict the structure service life. Another objective has been to study the effect of mineral additions in concrete and how they improve the durability of concrete in this environment. Efficiency factors have been obtained for fly ashes and silica fume, much higher than those consider for strength. According to the Spanish Concrete Code EHE 08, the atmospheric marine environment includes structures located at the cost, from the sea line up to 5km inland. The research conducted has shown the need to consider a spray zone which is the most unfavourable area of the atmospheric environment, affecting structures over the sea level as the decks of the wharves here studied. Surface chloride contents of structures in the different marine environments have been deeply analysed, checking the actual standard limits. As a conclusion, new normative values have been proposed for submerged and atmospheric zone (spray zone). A complete analysis of the chloride threshold concentration from the data obtained in real structures has also been performed. It has been evaluated the resistivity test as a tool to estimate the chloride diffusion coefficient. This test has proven effectiveness to discriminate concrete with mineral additions and low water penetration values, while it is not sensitive to detect concretes with high permeability. Finally, a chloride diffusion model (w/c) has been obtained in atmospheric marine environment in order to estimate the chloride diffusion of the concrete knowing a normative requirement as w/c.

La Península Ibérica en el transporte masivo de mercancías entre Europa y África : futuras autopistas del mar

La competitividad del transporte de mercancías depende del estado y funcionamiento de las redes existentes y de sus infraestructuras, no del modo de transporte. En concreto, la rentabilidad o la reducción de los costes de producción del transporte marítimo se vería incrementado con el uso de buques de mayor capacidad y con el desarrollo de plataformas portuarias de distribución o puertos secos, ya que el 90% del comercio entre la Unión Europea y terceros países se realiza a través de sus puertos a un promedio de 3,2 billones de toneladas de mercancías manipuladas cada año y el 40% del tráfico intraeuropeo utiliza el transporte marítimo de corta distancia. A pesar de que los puertos europeos acogen anualmente a más de 400 millones de pasajeros, los grandes desarrollos se han producido en los puertos del norte de Europa (Róterdam, Amberes, Ámsterdam). Los países del Sur de Europa deben buscar nuevas fórmulas para ser más competitivos, ya sea mediante creación de nuevas infraestructuras o mediante refuerzo de las existentes, ofreciendo los costes de los puertos del Norte. El fomento del transporte marítimo y fluvial como alternativa al transporte por carretera, especialmente el transporte marítimo de corta distancia, ha sido impulsado por la Comisión Europea (CE) desde 2003 a través de programas de apoyo comunitario de aplicación directa a las Autopistas del Mar, a modo de ejemplo, cabría citar los programas Marco Polo I y II, los cuales contaron con una dotación presupuestaria total de 855 millones de euros para el período 2003 – 2013; en ese período de tiempo se establecieron objetivos de reducción de congestión vial y mejora del comportamiento medio ambiental del sistema de transporte de mercancías dentro de la comunidad y la potenciación de la intermodalidad. El concepto de Autopista del Mar surge en el Libro Blanco de Transportes de la Comisión Europea “La política europea de transportes de cara al 2010: La hora de la verdad” del 12 de diciembre de 2001, en el marco de una política europea para fomento y desarrollo de sistemas de transportes sostenibles. Las Autopistas del Mar consisten en rutas marítimas de corta distancia entre dos puntos, de menor distancia que por vía terrestre, en las que a través del transporte intermodal mejoran significativamente los tiempos y costes de la cadena logística, contribuyen a la reducción de accidentes, ruidos y emisiones de CO2 a la atmósfera, permite que los conductores pierdan horas de trabajo al volante y evita el deterioro de las infraestructuras terrestres, con el consiguiente ahorro en mantenimiento. La viabilidad de una Autopista del Mar depende tanto de factores de ubicación geográficos, como de características propias del puerto, pasando por los diferentes requerimientos del mercado en cada momento (energéticos, medio ambientales y tecnológicos). Existe un elemento nuevo creado por la Comisión Europea: la red transeuropea de transportes (RTE-T). En el caso de España, con sus dos accesos por los Pirineos (La Junquera e Irún) como únicos pasos terrestres de comunicación con el continente y con importantes limitaciones ferroviarias debido a los tres anchos de vía distintos, le resta competitividad frente al conjunto europeo; por el contrario, España es el país europeo con más kilómetros de costa (con más de 8.000 km) y con un emplazamiento geográfico estratégico, lo que le convierte en una plataforma logística para todo el sur de Europa, por lo que las Autopistas del Mar tendrán un papel importante y casi obligado para el desarrollo de los grandes corredores marítimos que promueve Europa. De hecho, Gijón y Vigo lo han hecho muy bien con sus respectivas líneas definidas como Autopistas del Mar y que conectan con el puerto francés de Nantes-Saint Nazaire, ya que desde ahí los camiones pueden coger rutas hacia el Norte. Paralelamente, la Unión Europea ha iniciado los pasos para el impulso de la primera Autopista del Mar que conectará España con el mercado de Reino Unido, concretamente los Puertos de Bilbao y Tilbury. Además, España e Italia sellaron un acuerdo internacional para desarrollar Autopistas del Mar entre ambos países, comprometiéndose a impulsar una docena de rutas entre puertos del litoral mediterráneo español y el italiano. Actualmente, están en funcionando los trayectos como Barcelona-Génova, Valencia-Civitavecchia y Alicante- Nápoles, notablemente más cortos por mar que por carretera. Bruselas identificó cuatro grandes corredores marítimos que podrían concentrar una alta densidad de tráfico de buques, y en dos de ellos España ya tenía desde un principio un papel crucial. La Comisión diseñó el 14 de abril de 2004, a través del proyecto West-Mos, una red de tráfico marítimo que tiene como vías fundamentales la denominada Autopista del Báltico (que enlaza Europa central y occidental con los países bálticos), la Autopista de Europa suroriental (que une el Adriático con el Jónico y el Mediterráneo más oriental) y también la Autopista de Europa occidental y la Autopista de Europa suroccidental (que enlazan España con Reino Unido y la Francia atlántica y con la Francia mediterránea e Italia, respectivamente). Para poder establecer Autopistas del Mar entre la Península Ibérica y el Norte de Europa primará especialmente la retirada de camiones en la frontera pirenaica, donde el tráfico pesado tiene actualmente una intensidad media diaria de 8.000 unidades, actuando sobre los puntos de mayor congestión, como por ejemplo los Alpes, los Pirineos, el Canal de la Mancha, las carreteras fronterizas de Francia y Euskadi, y proponiendo el traslado de las mercancías en barcos o en trenes. Por su parte, para contar con los subsidios y apoyos europeos las rutas seleccionadas como Autopistas del Mar deben mantener una serie de criterios de calidad relacionados con la frecuencia, coste “plataforma logística a plataforma logística”, simplicidad en procedimientos administrativos y participación de varios países, entre otros. Los estudios consideran inicialmente viables los tramos marítimos superiores a 450 millas, con un volumen de unas 15.000 plataformas al año y que dispongan de eficientes comunicaciones desde el puerto a las redes transeuropeas de autopistas y ferrocarril. Otro objetivo de las Autopistas del Mar es desarrollar las capacidades portuarias de forma que se puedan conectar mejor las regiones periféricas a escala del continente europeo. En lo que a Puertos se refiere, las terminales en los muelles deben contar con una línea de atraque de 250 m., un calado superior a 8 m., una rampa “ro-ro” de doble calzada, grúas portainer, y garantizar operatividad para un mínimo de dos frecuencias de carga semanales. El 28 de marzo de 2011 se publicó el segundo Libro Blanco sobre el futuro del transporte en Europa “Hoja de ruta hacia un espacio único europeo de transporte: por una política de transportes competitiva y sostenible”, donde se definió el marco general de las acciones a emprender en los próximos diez años en el ámbito de las infraestructuras de transporte, la legislación del mercado interior, la reducción de la dependencia del carbono, la tecnología para la gestión del tráfico y los vehículos limpios, así como la estandarización de los distintos mercados. Entre los principales desafíos se encuentran la eliminación de los cuellos de botella y obstáculos diversos de nuestra red europea de transporte, minimizar la dependencia del petróleo, reducir las emisiones de GEI en un 60% para 2050 con respecto a los niveles de 1990 y la inversión en nuevas tecnologías e infraestructuras que reduzcan estas emisiones de transporte en la UE. La conexión entre la UE y el norte de África provoca elevados niveles de congestión en los puntos más críticos del trayecto: frontera hispano-francesa, corredor del Mediterráneo y el paso del estrecho. A esto se le añade el hecho de que el sector del transporte por carretera está sujeto a una creciente competencia de mercado motivada por la eliminación de las barreras europeas, mayores exigencias de los cargadores, mayores restricciones a los conductores y aumento del precio del gasóleo. Por otro lado, el mercado potencial de pasajeros tiene una clara diferenciación en tipos de flujos: los flujos en el período extraordinario de la Operación Paso del Estrecho (OPE), enfocado principalmente a marroquíes que vuelven a su país de vacaciones; y los flujos en el período ordinario, enfocado a la movilidad global de la población. Por tanto, lo que se pretende conseguir con este estudio es analizar la situación actual del tráfico de mercancías y pasajeros con origen o destino la península ibérica y sus causas, así como la investigación de las ventajas de la creación de una conexión marítima (Autopista del Mar) con el Norte de África, basándose en los condicionantes técnicos, administrativos, económicos, políticos, sociales y medio ambientales. The competitiveness of freight transport depends on the condition and operation of existing networks and infrastructure, not the mode of transport. In particular, profitability could be increased or production costs of maritime transport could be reduced by using vessels with greater capacity and developing port distribution platforms or dry ports, seeing as 90% of trade between the European Union and third countries happens through its ports. On average 3,2 billion tonnes of freight are handled annualy and 40% of intra-European traffic uses Short Sea Shipping. In spite of European ports annually hosting more than 400 million passengers, there have been major developments in the northern European ports (Rotterdam, Antwerp, Amsterdam). Southern European countries need to find new ways to be more competitive, either by building new infrastructure or by strengthening existing infrastructure, offering costs northern ports. The use of maritime and river transport as an alternative to road transport, especially Short Sea Shipping, has been driven by the European Commission (EC) from 2003 through community support programs for the Motorways of the Sea. These programs include, for example, the Marco Polo I and II programs, which had a total budget of 855 million euros for the period 2003-2013. During this time objectives were set for reducing road congestion, improving the environmental performance of the freight transport system within the community and enhancing intermodal transport. The “Motorway of the Sea” concept arises in the European Commission’s Transport White Paper "European transport policy for 2010: time to decide" on 12 December 2001, as part of a European policy for the development and promotion of sustainable transport systems. A Motorway of the Sea is defined as a short sea route between two points, covering less distance than by road, which provides a significant improvement in intermodal transport times and to the cost supply chain. It contributes to reducing accidents, noise and CO2 emissions, allows drivers to shorten their driving time and prevents the deterioration of land infrastructure thereby saving on maintenance costs. The viability of a Motorway of the Sea depends as much on geographical location factors as on characteristics of the port, taking into account the different market requirements at all times (energy, environmental and technological). There is a new element created by the European Commission: the trans-European transport network (TEN-T). In the case of Spain, with its two access points in the Pyrenees (La Junquera and Irun) as the only land crossings connected to the mainland and major railway limitations due to the three different gauges, it appears less competitive compared to Europe as a whole. However, Spain is the European country with the most kilometers of coastline (over 8,000 km) and a strategic geographical location, which makes it a logistics platform for the all of Southern Europe. This is why the Motorways of the Sea will have an important role, and an almost necessary one to develop major maritime corridors that Europe supports. In fact, Gijon and Vigo have done very well with their respective sea lanes defined as Motorways of the Sea and which connect with the French port of Nantes-Saint Nazaire, as from there trucks can use nort-heading routes. In parallel, the European Union has taken the first steps to boost the first Motorway of the Sea linking Spain to the UK market, specifically the ports of Bilbao and Tilbury. Furthermore, Spain and Italy sealed an international agreement to develop Motorways of the Sea between both countries, pledging to develop a dozen routes between ports on the Spanish and Italian Mediterranean coasts. Currently, there are sea lanes already in use such as Barcelona-Genova, Valencia-Civitavecchia and Alicante-Naples, these are significantly shorter routes by sea than by road. Brussels identified four major maritime corridors that could hold heavy concentrate shipping traffic, and Spain had a crucial role in two of these from the beginning. On 14 April 2004 the Commission planned through the West-Mos project, a network of maritime traffic which includes the essential sea passages the so-called Baltic Motorway (linking Central and Western Europe with the Baltic countries), the southeast Europe Motorway (linking the Adriatic to the Ionian and eastern Mediterranean Sea), the Western Europe Motorway and southwestern Europe Motorway (that links Spain with Britain and the Atlantic coast of France and with the French Mediterranean coast and Italy, respectively). In order to establish Motorways of the Sea between the Iberian Peninsula and Northern Europe especially, it is necessary to remove trucks from the Pyrenean border, where sees heavy traffic (on average 8000 trucks per day) and addressing the points of greatest congestion, such as the Alps, the Pyrenees, the English Channel, the border roads of France and Euskadi, and proposing the transfer of freight on ships or trains. For its part, in order to receive subsidies and support from the European Commission, the routes selected as Motorways of the Sea should maintain a series of quality criteria related to frequency, costs "from logistics platform to logistics platform," simplicity in administrative procedures and participation of several countries, among others. To begin with, studies consider viable a maritime stretch of at least 450 miles with a volume of about 15,000 platforms per year and that have efficient connections from port to trans-European motorways and rail networks. Another objective of the Motorways of the Sea is to develop port capacity so that they can better connect peripheral regions across the European continent. Referring ports, the terminals at the docks must have a berthing line of 250 m., a draft greater than 8 m, a dual carriageway "ro-ro" ramp, portainer cranes, and ensure operability for a minimum of two loads per week. On 28 March 2011 the second White Paper about the future of transport in Europe "Roadmap to a Single European Transport Area – Towards a competitive and resource efficient transport system" was published. In this Paper the general framework of actions to be undertaken in the next ten years in the field of transport infrastructure was defined, including internal market legislation, reduction of carbon dependency, traffic management technology and clean vehicles, as well as the standardization of different markets. The main challenges are how to eliminate bottlenecks and various obstacles in our European transport network, minimize dependence on oil, reduce GHG emissions by 60% by 2050 compared to 1990 levels and encourage investment in new technologies and infrastructure that reduce EU transport emissions. The connection between the EU and North Africa causes high levels of congestion on the most critical points of the journey: the Spanish-French border, the Mediterranean corridor and Gibraltar Strait. In addition to this, the road transport sector is subject to increased market competition motivated by the elimination of European barriers, greater demands of shippers, greater restrictions on drivers and an increase in the price of diesel. On the other hand, the potential passenger market has a clear differentiation in type of flows: flows in the special period of the Crossing the Straits Operation (CSO), mainly focused on Moroccans who return home on vacation; and flows in the regular session, focused on the global mobile population. Therefore, what I want to achieve with this study is present an analysis of the current situation of freight and passengers to or from the Iberian Peninsula and their causes, as well as present research on the advantages of creating a maritime connection (Motorways of the Sea) with North Africa, based on the technical, administrative, economic, political, social and environmental conditions.

The small ribosomal subunit from Thermus thermophilus at 4.5 Å resolution: Pattern fittings and the identification of a functional site

The electron density map of the small ribosomal subunit from Thermus thermophilus, constructed at 4.5 Å resolution, shows the recognizable morphology of this particle, as well as structural features that were interpreted as ribosomal RNA and proteins. Unbiased assignments, carried out by quantitative covalent binding of heavy atom compounds at predetermined sites, led to the localization of the surface of the ribosomal protein S13 at a position compatible with previous assignments, whereas the surface of S11 was localized at a distance of about twice its diameter from the site suggested for its center by neutron scattering. Proteins S5 and S7, whose structures have been determined crystallographically, were visually placed in the map with no alterations in their conformations. Regions suitable to host the fold of protein S15 were detected in several positions, all at a significant distance from the location of this protein in the neutron scattering map. Targeting the 16S RNA region, where mRNA docks to allow the formation of the initiation complex by a mercurated mRNA analog, led to the characterization of its vicinity.

The myosin motor, Myo4p, binds Ash1 mRNA via the adapter protein, She3p

In Saccharomyces cerevisiae, mRNA encoding the cell-fate determinant Ash1p is localized to the distal tip of daughter cells. Five SHE genes are required for proper Ash1 mRNA localization, one of which encodes the myosin Myo4p. We show that three of the five She proteins, She2p, She3p, and Myo4p, colocalize with Ash1 mRNA in vivo and coimmunoprecipitate with Ash1 mRNA from cell extracts. We also find that She3p binds to Myo4p in the absence of RNA and She2p is required for binding She3p-Myo4p to Ash1 mRNA. These results suggest that She3p acts as an adapter protein that docks the myosin motor onto an Ash1–She2p ribonucleoprotein complex.

Dunkerque, France, 1695 (Raster Image)

This layer is a georeferenced raster image of the historic paper map entitled: Plan en profil van Duynkerken : met sijn sterktens en zee-kasteelen, belegert te water, engebombardeert door de Engelse en Hollantse zee-magten, onder de Engelse admiraal Berkly en Hollantse admiraal Alemonde, op den Augustus, 1695, doen maken en uytgeven na de autentijke tekeninge van Monsieur Pamer, Ingenieur van Sijn Kon. Majest. van Groot Brittannien. It was published by Pieter Persoy, op den Dam in 1695. Scale [ca. 1:13,000]. Covers a portion of Dunkerque, France. Map in Dutch. The image inside the map neatline is georeferenced to the surface of the earth and fit to the European Datum 1950, Universal Transverse Mercator (UTM) Zone 31N projected coordinate system. All map collar and inset information is also available as part of the raster image, including any inset maps, profiles, statistical tables, directories, text, illustrations, index maps, legends, or other information associated with the principal map. This map shows features such as roads, drainage, built-up areas and selected buildings, fortifications and defenses, canals, docks, ground cover, and more. Includes also index and panorama view.This layer is part of a selection of digitally scanned and georeferenced historic maps from the Harvard Map Collection. These maps typically portray both natural and manmade features. The selection represents a range of originators, ground condition dates, scales, and map purposes.

Le Havre, Haute-Normandie, France, ca. 1880 (Raster Image)

This layer is a georeferenced raster image of the historic paper map entitled: Plan de la ville & du Port du Havre. It was published by Eug. Costey, éditeur ca. 1880. Scale [ca. 1:16,400]. Covers Le Havre region, Haute-Normandie, France. Map in French. The image inside the map neatline is georeferenced to the surface of the earth and fit to the European Datum 1950, Universal Transverse Mercator (UTM) Zone 31N projected coordinate system. All map collar and inset information is also available as part of the raster image, including any inset maps, profiles, statistical tables, directories, text, illustrations, index maps, legends, or other information associated with the principal map. This map shows features such as roads, railroads and stations, drainage, selected buildings, parks, cemeteries, ground cover, city districts, fortification, canals, wharves, docks, and more.This layer is part of a selection of digitally scanned and georeferenced historic maps from the Harvard Map Collection. These maps typically portray both natural and manmade features. The selection represents a range of originators, ground condition dates, scales, and map purposes.

Exposition universelle d'Anvers, Antwerp, Belgium, 1885 (Raster Image)

This layer is a georeferenced raster image of the historic paper map entitled: Plan instantané de la ville d'Anvers : indiquant l'Exposition Universelle et tous les principaux monuments en relief, le tracé des tramways, les nouveaux quais hangars, etc., etc., dressé par E.V.D.H. It was published by Richd. Huybrechts & Cie in 1885. Scale [ca. 1:11,000]. Covers Antwerp, Belgium. Map in French. The image inside the map neatline is georeferenced to the surface of the earth and fit to the Belgian Lambert 1972 coordinate system. All map collar and inset information is also available as part of the raster image, including any inset maps, profiles, statistical tables, directories, text, illustrations, index maps, legends, or other information associated with the principal map. This map shows features such as roads, railroads, drainage, built-up areas and selected buildings, ground cover, fortifications, canals, docks, and more. Relief shown pictorially. Shows buildings and points of interest related to the Exposition universelle d'Anvers.This layer is part of a selection of digitally scanned and georeferenced historic maps from the Harvard Map Collection. These maps typically portray both natural and manmade features. The selection represents a range of originators, ground condition dates, scales, and map purposes.

Nice, France, 1879 (Raster Image)

This layer is a georeferenced raster image of the historic paper map entitled: Plan pittoresque de Nice, par un touriste. It was published by Lith. J. Dalmet in 1879. Scale [ca. 1:7,353]. Covers Nice, France. Map in French.The image inside the map neatline is georeferenced to the surface of the earth and fit to the European Datum 1950, Universal Transverse Mercator (UTM) Zone 32N projected coordinate system. All map collar and inset information is also available as part of the raster image, including any inset maps, profiles, statistical tables, directories, text, illustrations, index maps, legends, or other information associated with the principal map. This map shows features such as roads, railroads and stations, drainage, built-up areas and selected buildings (some shown pictorially), industry locations, docks and wharves, gardens, and more. Includes two insets: "Jardin d'Acclimatation de la Société d'Agriculture des Alpes-Maritimes" and "Nice à Villefranche, Saint Jean et Beaulieu"; and a view of the "Villa Chauvain à Villefranche".This layer is part of a selection of digitally scanned and georeferenced historic maps from the Harvard Map Collection. These maps typically portray both natural and manmade features. The selection represents a range of originators, ground condition dates, scales, and map purposes.