Do international flights promote FDI? : the role of face-to-face communication

Air transportation facilitates face-to-face interactions across borders for the spatial expansion of manufacturing production. I investigate the impact of international flights on FDI entry by Japanese firms. I find that FDI entry significantly increases with the weekly frequency of flights from Japan, and the positive impact increases with a proxy for an intensity of face-to-face communication between the parent firm and foreign affiliate. The results are robust to estimation methods, additional control variables, and definitions of face-to-face communication. Thus, the results suggest that flights encourage FDI entry through a reduction in face-to-face communication costs.

Is the concept of a green economy a useful way of framing Policy Discussions and policymaking to promote sustainable development?

In the Viewpoints section, academics, practitioners and experts share their perspectives on policy questions relevant to sustainable development. In this issue, experts address the question: “Is the concept of a green economy a useful way of framing policy discussions and policymaking to promote sustainable development?

Work in progress - Exploiting videoconferencing possibilities to promote the European convergence process

This paper introduces the experience of using videoconferencing and recording as a mechanism to support courses which need to be promoted or discontinued within the framework of the European convergence process. Our objective is to make these courses accessible as live streaming during the lessons as well as recorded lectures and associated documents available to the students as soon as the lesson has finished. The technology used has been developed in our university and it is all open source. Although this is a technical project the key is the human factor involved. The people managing the virtual sessions are students of the courses being recorded. However, they lack technical knowledge, so we had to train them in audiovisuals and enhance the usability of the videoconferencing tool and platform. The validation process is being carried out in five real scenarios at our university. During the whole period we are evaluating technical and pedagogical issues of this experience for both students and teachers to guide the future development of the service. Depending on the final results, the service of lectures recording will be available as educational resource for all of the teaching staff of our university.

Using multimedia and peer assessment to promote collaborative e-learning

Collaborative e-learning is increasingly appealing as a pedagogical approach that can positively affect student learning. We propose a didactical model that integrates multimedia with collaborative tools and peer assessment to foster collaborative e-learning. In this paper, we explain it and present the results of its application to the “International Seminars on Materials Science” online course. The proposed didactical model consists of five educational activities. In the first three, students review the multimedia resources proposed by the teacher in collaboration with their classmates. Then, in the last two activities, they create their own multimedia resources and assess those created by their classmates. These activities foster communication and collaboration among students and their ability to use and create multimedia resources. Our purpose is to encourage the creativity, motivation, and dynamism of the learning process for both teachers and students.

Measurements and Prediction of Pedestrian Walking Loads on an Aluminium Catwalk

During the last two decades the topic of human induced vibration has attracted a lot of attention among civil engineering practitioners and academics alike. Usually this type of problem may be encountered in pedestrian footbridges or floors of paperless offices. Slender designs are becoming increasingly popular, and as a consequence, the importance of paying attention to vibration serviceability also increases. This paper resumes the results obtained from measurements taken at different points of an aluminium catwalk which is 6 m in length by 0.6 m in width. Measurements were carried out when subjecting the structure to different actions:1)Static test: a steel cylinder of 35 kg was placed in the middle of the catwalk; 2)Dynamic test: this test consists of exciting the structure with singles impulses; 3)Dynamic test: people walking on the catwalk. Identification of the mechanical properties of the structure is an achievement of the paper. Indirect methods were used to estimate properties including the support stiffness, the beam bending stiffness, the mass of the structure (using Rayleigh method and iterative matrix method), the natural frequency (using the time domain and frequency domain analysis) and the damping ratio (by calculating the logarithmic decrement). Experimental results and numerical predictions for the response of an aluminium catwalk subjected to walking loads have been compared. The damping of this light weight structure depends on the amplitude of vibration which complicates the tuning of a structural model. In the light of the results obtained it seems that the used walking load model is not appropriate as the predicted transient vibration values (TTVs) are much higher than the measured ones.

EU support mechanisms to promote public private partnerships for financing Trans-European transport infrastructure

Since the mid 80ies the Trans ‐European Transport Network (TEN‐T) policy has been setting the framework for the development of infrastructure for the smooth functioning of the internal market within the European Union (EU). Public Private Partnerships (PPPs) have always been regarded by the EU as a key instrument to promote the TEN‐T.

Utilizing the Internet of Things to promote energy awareness and efficiency at discrete production processes: Practices and methodology

El actual contexto de fabricación, con incrementos en los precios de la energía, una creciente preocupación medioambiental y cambios continuos en los comportamientos de los consumidores, fomenta que los responsables prioricen la fabricación respetuosa con el medioambiente. El paradigma del Internet de las Cosas (IoT) promete incrementar la visibilidad y la atención prestada al consumo de energía gracias tanto a sensores como a medidores inteligentes en los niveles de máquina y de línea de producción. En consecuencia es posible y sencillo obtener datos de consumo de energía en tiempo real proveniente de los procesos de fabricación, pero además es posible analizarlos para incrementar su importancia en la toma de decisiones. Esta tesis pretende investigar cómo utilizar la adopción del Internet de las Cosas en el nivel de planta de producción, en procesos discretos, para incrementar la capacidad de uso de la información proveniente tanto de la energía como de la eficiencia energética. Para alcanzar este objetivo general, la investigación se ha dividido en cuatro sub-objetivos y la misma se ha desarrollado a lo largo de cuatro fases principales (en adelante estudios). El primer estudio de esta tesis, que se apoya sobre una revisión bibliográfica comprehensiva y sobre las aportaciones de expertos, define prácticas de gestión de la producción que son energéticamente eficientes y que se apoyan de un modo preeminente en la tecnología IoT. Este primer estudio también detalla los beneficios esperables al adoptar estas prácticas de gestión. Además, propugna un marco de referencia para permitir la integración de los datos que sobre el consumo energético se obtienen en el marco de las plataformas y sistemas de información de la compañía. Esto se lleva a cabo con el objetivo último de remarcar cómo estos datos pueden ser utilizados para apalancar decisiones en los niveles de procesos tanto tácticos como operativos. Segundo, considerando los precios de la energía como variables en el mercado intradiario y la disponibilidad de información detallada sobre el estado de las máquinas desde el punto de vista de consumo energético, el segundo estudio propone un modelo matemático para minimizar los costes del consumo de energía para la programación de asignaciones de una única máquina que deba atender a varios procesos de producción. Este modelo permite la toma de decisiones en el nivel de máquina para determinar los instantes de lanzamiento de cada trabajo de producción, los tiempos muertos, cuándo la máquina debe ser puesta en un estado de apagada, el momento adecuado para rearrancar, y para pararse, etc. Así, este modelo habilita al responsable de producción de implementar el esquema de producción menos costoso para cada turno de producción. En el tercer estudio esta investigación proporciona una metodología para ayudar a los responsables a implementar IoT en el nivel de los sistemas productivos. Se incluye un análisis del estado en que se encuentran los sistemas de gestión de energía y de producción en la factoría, así como también se proporcionan recomendaciones sobre procedimientos para implementar IoT para capturar y analizar los datos de consumo. Esta metodología ha sido validada en un estudio piloto, donde algunos indicadores clave de rendimiento (KPIs) han sido empleados para determinar la eficiencia energética. En el cuarto estudio el objetivo es introducir una vía para obtener visibilidad y relevancia a diferentes niveles de la energía consumida en los procesos de producción. El método propuesto permite que las factorías con procesos de producción discretos puedan determinar la energía consumida, el CO2 emitido o el coste de la energía consumida ya sea en cualquiera de los niveles: operación, producto o la orden de fabricación completa, siempre considerando las diferentes fuentes de energía y las fluctuaciones en los precios de la misma. Los resultados muestran que decisiones y prácticas de gestión para conseguir sistemas de producción energéticamente eficientes son posibles en virtud del Internet de las Cosas. También, con los resultados de esta tesis los responsables de la gestión energética en las compañías pueden plantearse una aproximación a la utilización del IoT desde un punto de vista de la obtención de beneficios, abordando aquellas prácticas de gestión energética que se encuentran más próximas al nivel de madurez de la factoría, a sus objetivos, al tipo de producción que desarrolla, etc. Así mismo esta tesis muestra que es posible obtener reducciones significativas de coste simplemente evitando los períodos de pico diario en el precio de la misma. Además la tesis permite identificar cómo el nivel de monitorización del consumo energético (es decir al nivel de máquina), el intervalo temporal, y el nivel del análisis de los datos son factores determinantes a la hora de localizar oportunidades para mejorar la eficiencia energética. Adicionalmente, la integración de datos de consumo energético en tiempo real con datos de producción (cuando existen altos niveles de estandarización en los procesos productivos y sus datos) es esencial para permitir que las factorías detallen la energía efectivamente consumida, su coste y CO2 emitido durante la producción de un producto o componente. Esto permite obtener una valiosa información a los gestores en el nivel decisor de la factoría así como a los consumidores y reguladores. ABSTRACT In today‘s manufacturing scenario, rising energy prices, increasing ecological awareness, and changing consumer behaviors are driving decision makers to prioritize green manufacturing. The Internet of Things (IoT) paradigm promises to increase the visibility and awareness of energy consumption, thanks to smart sensors and smart meters at the machine and production line level. Consequently, real-time energy consumption data from the manufacturing processes can be easily collected and then analyzed, to improve energy-aware decision-making. This thesis aims to investigate how to utilize the adoption of the Internet of Things at shop floor level to increase energy–awareness and the energy efficiency of discrete production processes. In order to achieve the main research goal, the research is divided into four sub-objectives, and is accomplished during four main phases (i.e., studies). In the first study, by relying on a comprehensive literature review and on experts‘ insights, the thesis defines energy-efficient production management practices that are enhanced and enabled by IoT technology. The first study also explains the benefits that can be obtained by adopting such management practices. Furthermore, it presents a framework to support the integration of gathered energy data into a company‘s information technology tools and platforms, which is done with the ultimate goal of highlighting how operational and tactical decision-making processes could leverage such data in order to improve energy efficiency. Considering the variable energy prices in one day, along with the availability of detailed machine status energy data, the second study proposes a mathematical model to minimize energy consumption costs for single machine production scheduling during production processes. This model works by making decisions at the machine level to determine the launch times for job processing, idle time, when the machine must be shut down, ―turning on‖ time, and ―turning off‖ time. This model enables the operations manager to implement the least expensive production schedule during a production shift. In the third study, the research provides a methodology to help managers implement the IoT at the production system level; it includes an analysis of current energy management and production systems at the factory, and recommends procedures for implementing the IoT to collect and analyze energy data. The methodology has been validated by a pilot study, where energy KPIs have been used to evaluate energy efficiency. In the fourth study, the goal is to introduce a way to achieve multi-level awareness of the energy consumed during production processes. The proposed method enables discrete factories to specify energy consumption, CO2 emissions, and the cost of the energy consumed at operation, production and order levels, while considering energy sources and fluctuations in energy prices. The results show that energy-efficient production management practices and decisions can be enhanced and enabled by the IoT. With the outcomes of the thesis, energy managers can approach the IoT adoption in a benefit-driven way, by addressing energy management practices that are close to the maturity level of the factory, target, production type, etc. The thesis also shows that significant reductions in energy costs can be achieved by avoiding high-energy price periods in a day. Furthermore, the thesis determines the level of monitoring energy consumption (i.e., machine level), the interval time, and the level of energy data analysis, which are all important factors involved in finding opportunities to improve energy efficiency. Eventually, integrating real-time energy data with production data (when there are high levels of production process standardization data) is essential to enable factories to specify the amount and cost of energy consumed, as well as the CO2 emitted while producing a product, providing valuable information to decision makers at the factory level as well as to consumers and regulators.

Analysis and optimization for a hexapod walking robot for planetary missions

La tecnología de las máquinas móviles autónomas ha sido objeto de una gran investigación y desarrollo en las últimas décadas. En muchas actividades y entornos, los robots pueden realizar operaciones que son duras, peligrosas o simplemente imposibles para los humanos. La exploración planetaria es un buen ejemplo de un entorno donde los robots son necesarios para realizar las tareas requeridas por los científicos. La reciente exploración de Marte con robots autónomos nos ha mostrado la capacidad de las nuevas tecnologías. Desde la invención de la rueda, que esta acertadamente considerado como el mayor invento en la historia del transporte humano, casi todos los vehículos para exploración planetaria han empleado las ruedas para su desplazamiento. Las nuevas misiones planetarias demandan maquinas cada vez mas complejas. En esta Tesis se propone un nuevo diseño de un robot con patas o maquina andante que ofrecerá claras ventajas en entornos extremos. Se demostrara que puede desplazarse en los terrenos donde los robots con ruedas son ineficientes, convirtiéndolo en una elección perfecta para misiones planetarias. Se presenta una reseña histórica de los principales misiones espaciales, en particular aquellos dirigidos a la exploración planetaria. A través de este estudio será posible analizar las desventajas de los robots con ruedas utilizados en misiones anteriores. El diseño propuesto de robot con patas será presentado como una alternativa para aquellas misiones donde los robots con ruedas puedan no ser la mejor opción. En esta tesis se presenta el diseño mecánico de un robot de seis patas capaz de soportar las grandes fuerzas y momentos derivadas del movimiento de avance. Una vez concluido el diseño mecánico es necesario realizar un análisis que permita entender el movimiento y comportamiento de una maquina de esta complejidad. Las ecuaciones de movimiento del robot serán validadas por dos métodos: cinemático y dinámico. Dos códigos Matlab® han sido desarrollados para resolver dichos sistemas de ecuaciones y han sido verificados por un tercer método, un modelo de elementos finitos, que también verifica el diseño mecánico. El robot con patas presentado, ha sido diseñado para la exploración planetaria en Marte. El comportamiento del robot durante sus desplazamientos será probado mediante un código de Matlab®, desarrollado para esta tesis, que permite modificar las trayectorias, el tipo de terreno, y el número y altura de los obstáculos. Estos terrenos y requisitos iniciales no han sido elegidos de forma aleatoria, si no que están basados en mi experiencia como miembro del equipo de MSL-NASA que opera un instrumento a bordo del rover Curiosity en Marte. El robot con patas desarrollado y fabricado por el Centro de Astrobiología (INTA-CSIC), esta basado en el diseño mecánico y análisis presentados en esta tesis. ABSTRACT The autonomous machines technology has undergone a major research and development during the last decades. In many activities and environments, robots can perform operations that are tought, dangerous or simply imposible to humans. Planetary exploration is a good example of such environment where robots are needed to perform the tasks required by the scientits. Recent Mars exploration based on autonomous vehicles has shown us the capacity of the new technologies. From the invention of the wheel, which is rightly regarded as the greatest invention in the history of human transportation, nearly all-planetary vehicles are based in wheeled locomotion, but new missions demand new types of machines due to the complex tasks needed to be performed. It will be proposed in this thesis a new design of a legged robot or walking machine, which may offer clear advantages in tough environments. This Thesis will show that the proposed walking machine can travel, were terrain difficulties make wheeled vehicles ineffective, making it a perfect choice for planetary mission. A historical background of the main space missions, in particular those aimed at planetary exploration will be presented. From this study the disadvantages found in the existing wheel rovers will be analysed. The legged robot designed will be introduced as an alternative were wheeled rovers could be no longer the best option for planetary exploration. This thesis introduces the mechanical design of a six-leg robot capable of withstanding high forces and moments due to the walking motion. Once the mechanical design is concluded, and in order to analyse a machine of this complexity an understanding of its movement and behaviour is mandatory. This movement equation will be validated by two methods: kinematics and dynamics. Two Matlab® codes have been developed to solve the systems of equations and validated by a third method, a finite element model, which also verifies the mechanical design. The legged robot presented has been designed for a Mars planetary exploration. The movement behaviour of the robot will be tested in a Matlab® code developed that allows to modify the trajectories, the type of terrain, number and height of obstacles. These terrains and initial requirements have not been chosen randomly, those are based on my experience as a member of the MSL NASA team, which operates an instrument on-board of the Curiosity rover in Mars. The walking robot developed and manufactured by the Center of Astrobiology (CAB) is based in the mechanical design and analysis that will be presented in this thesis.

Why do health economists promote technology adoption rather than the search for efficiency? A proposal for a change in our approach to economic evaluation in health care

Acknowledgments Financial Support: HERU and HSRU receive a core grant from the Chief Scientist’s Office of the Scottish Government Health and Social Care Directorates, and the Centre for Clinical epidemiology & Evaluation is funded by Vancouver Coastal Health Authority. The model used for the illustrative case study in this paper was developed as part of a NHS Technology Assessment Review, funded by the National Institute for Health Research (NIHR) Health Technology Assessment Program (project number 09/146/01). The views and opinions expressed in this paper are those of the authors and do not necessarily reflect those of the Scottish Government, NHS, Vancouver Coastal Health, NIHR HTA Program or the Department of Health. The authors wish to thank Kathleen Boyd and members of the audience at the UK Health Economists Study Group, for comments received on an earlier version of this paper. We also wish to thank Cynthia Fraser (University of Aberdeen) for literature searches undertaken to inform the manuscript, and Mohsen Sadatsafavi (University of British Columbia) for comments on an earlier draft

CM101-mediated recovery of walking ability in adult mice paralyzed by spinal cord injury

CM101, an antiangiogenic polysaccharide derived from group B streptococcus, was administered by i.v. injection 1 hr post-spinal-cord crush injury in an effort to prevent inflammatory angiogenesis and gliosis (scarring) in a mouse model. We postulated that gliosis would sterically prevent the reestablishment of neuronal connectivity; thus, treatment with CM101 was repeated every other day for five more infusions for the purpose of facilitating regeneration of neuronal function. Twenty-five of 26 mice treated with CM101 survived 28 days after surgery, and 24 of 26 recovered walking ability within 2–12 days. Only 6 of 14 mice in the control groups survived 24 hr after spinal cord injury, and none recovered function in paralyzed limbs. MRI analysis of injured untreated and treated animals showed that CM101 reduced the area of damage at the site of spinal cord compression, which was corroborated by histological analysis of spinal cord sections from treated and control animals. Electrophysiologic measurements on isolated central nervous system and neurons in culture showed that CM101 protected axons from Wallerian degeneration; reversed γ-aminobutyrate-mediated depolarization occurring in traumatized neurons; and improved recovery of neuronal conductivity of isolated central nervous system in culture.

Endothelial selectins and vascular cell adhesion molecule-1 promote hematopoietic progenitor homing to bone marrow

The adhesive mechanisms allowing hematopoietic progenitor cells (HPC) homing to the bone marrow (BM) after BM transplantation are poorly understood. We investigated the role of endothelial selectins and vascular cell adhesion molecule-1 (VCAM-1) in this process. Lethally irradiated recipient mice deficient in both P-and E-selectins (P/E−/−), reconstituted with minimal numbers (≤5 × 104) of wild-type BM cells, poorly survived the procedure compared with wild-type recipients. Excess mortality in P/E−/− mice, after a lethal dose of irradiation, was likely caused by a defect of HPC homing. Indeed, we observed that the recruitment of HPC to the BM was reduced in P/E−/− animals, either splenectomized or spleen-intact. Homing into the BM of P/E−/− recipient mice was further compromised when a function-blocking VCAM-1 antibody was administered. Circulating HPC, 14 hr after transplantation, were greatly increased in P/E−/− mice treated with anti-VCAM-1 compared with P/E−/− mice treated with just IgG or wild-type mice treated with either anti-VCAM-1 or IgG. Our results indicate that endothelial selectins play an important role in HPC homing to the BM. Optimal recruitment of HPC after lethal doses of irradiation requires the combined action of both selectins and VCAM-1 expressed on endothelium of the BM.

X-ngnr-1 and Xath3 promote ectopic expression of sensory neuron markers in the neurula ectoderm and have distinct inducing properties in the retina

Xath3 encodes a Xenopus neuronal-specific basic helix–loop–helix transcription factor related to the Drosophila proneural factor atonal. We show here that Xath3 acts downstream of X-ngnr-1 during neuronal differentiation in the neural plate and retina and that its expression and activity are modulated by Notch signaling. X-ngnr-1 activates Xath3 and NeuroD by different mechanisms, and the latter two genes crossactivate each other. In the ectoderm, X-ngnr-1 and Xath3 have similar activities, inducing ectopic sensory neurons. Among the sensory-specific markers tested, only those that label cranial neurons were found to be ectopically activated. By contrast, in the retina, X-ngnr-1 and Xath3 overexpression promote the development of overlapping but distinct subtypes of retinal neurons. Together, these data suggest that X-ngnr-1 and Xath3 regulate successive stages of early neuronal differentiation and that, in addition to their general proneural properties, they may contribute, in a context-dependent manner, to some aspect of neuronal identity.

Virus-sized self-assembling lamellar complexes between plasmid DNA and cationic micelles promote gene transfer

Gene therapy is based on the vectorization of genes to target cells and their subsequent expression. Cationic amphiphile-mediated delivery of plasmid DNA is the nonviral gene transfer method most often used. We examined the supramolecular structure of lipopolyamine/plasmid DNA complexes under various condensing conditions. Plasmid DNA complexation with lipopolyamine micelles whose mean diameter was 5 nm revealed three domains, depending on the lipopolyamine/plasmid DNA ratio. These domains respectively corresponded to negatively, neutrally, and positively charged complexes. Transmission electron microscopy and x-ray scattering experiments on complexes originating from these three domains showed that although their morphology depends on the lipopolyamine/plasmid DNA ratio, their particle structure consists of ordered domains characterized by even spacing of 80 Å, irrespective of the lipid/DNA ratio. The most active lipopolyamine/DNA complexes for gene transfer were positively charged. They were characterized by fully condensed DNA inside spherical particles (diameter: 50 nm) sandwiched between lipid bilayers. These results show that supercoiled plasmid DNA is able to transform lipopolyamine micelles into a supramolecular organization characterized by ordered lamellar domains.

Cellulose-binding domains promote hydrolysis of different sites on crystalline cellulose

The cohesin-dockerin interaction in Clostridium thermocellum cellulosome mediates the tight binding of cellulolytic enzymes to the cellulosome-integrating protein CipA. Here, this interaction was used to study the effect of different cellulose-binding domains (CBDs) on the enzymatic activity of C. thermocellum endoglucanase CelD (1,4-β-d endoglucanase, EC3.2.1.4) toward various cellulosic substrates. The seventh cohesin domain of CipA was fused to CBDs originating from the Trichoderma reesei cellobiohydrolases I and II (CBDCBH1 and CBDCBH2) (1,4-β-d glucan-cellobiohydrolase, EC3.2.1.91), from the Cellulomonas fimi xylanase/exoglucanase Cex (CBDCex) (β-1,4-d glucanase, EC3.2.1.8), and from C. thermocellum CipA (CBDCipA). The CBD-cohesin hybrids interacted with the dockerin domain of CelD, leading to the formation of CelD-CBD complexes. Each of the CBDs increased the fraction of cellulose accessible to hydrolysis by CelD in the order CBDCBH1 < CBDCBH2 ≈ CBDCex < CBDCipA. In all cases, the extent of hydrolysis was limited by the disappearance of sites accessible to CelD. Addition of a batch of fresh cellulose after completion of the reaction resulted in a new burst of activity, proving the reversible binding of the intact complexes despite the apparent binding irreversibility of some CBDs. Furthermore, burst of activity also was observed upon adding new batches of CelD–CBD complexes that contained a CBD differing from the first one. This complementation between different CBDs suggests that the sites made available for hydrolysis by each of the CBDs are at least partially nonoverlapping. The only exception was CBDCipA, whose sites appeared to overlap all of the other sites.

Subgenome chromosome walking in wheat: A 450-kb physical contig in Triticum monococcum L. spans the Lr10 resistance locus in hexaploid wheat (Triticum aestivum L.)

For many agronomically important plant genes, only their position on a genetic map is known. In the absence of an efficient transposon tagging system, such genes have to be isolated by map-based cloning. In bread wheat Triticum aestivum, the genome is hexaploid, has a size of 1.6 × 1010 bp, and contains more than 80% of repetitive sequences. So far, this genome complexity has not allowed chromosome walking and positional cloning. Here, we demonstrate that chromosome walking using bacterial artificial chromosome (BAC) clones is possible in the diploid wheat Triticum monococcum (Am genome). BAC end sequences were mostly repetitive and could not be used for the first walking step. New probes corresponding to rare low-copy sequences were efficiently identified by low-pass DNA sequencing of the BACs. Two walking steps resulted in a physical contig of 450 kb on chromosome 1AmS. Genetic mapping of the probes derived from the BAC contig demonstrated perfect colinearity between the physical map of T. monococcum and the genetic map of bread wheat on chromosome 1AS. The contig genetically spans the Lr10 leaf rust disease resistance locus in bread wheat, with 0.13 centimorgans corresponding to 300 kb between the closest flanking markers. Comparison of the genetic to physical distances has shown large variations within 350 kb of the contig. The physical contig can now be used for the isolation of the orthologous regions in bread wheat. Thus, subgenome chromosome walking in wheat can produce large physical contigs and saturate genomic regions to support positional cloning.