Configuration management and product lines to enhance the replication process in software engineering

This research is concerned with the experimental software engineering area, specifically experiment replication. Replication has traditionally been viewed as a complex task in software engineering. This is possibly due to the present immaturity of the experimental paradigm applied to software development. Researchers usually use replication packages to replicate an experiment. However, replication packages are not the solution to all the information management problems that crop up when successive replications of an experiment accumulate. This research borrows ideas from the software configuration management and software product line paradigms to support the replication process. We believe that configuration management can help to manage and administer information from one replication to another: hypotheses, designs, data analysis, etc. The software product line paradigm can help to organize and manage any changes introduced into the experiment by each replication. We expect the union of the two paradigms in replication to improve the planning, design and execution of further replications and their alignment with existing replications. Additionally, this research work will contribute a web support environment for archiving information related to different experiment replications. Additionally, it will provide flexible enough information management support for running replications with different numbers and types of changes. Finally, it will afford massive storage of data from different replications. Experimenters working collaboratively on the same experiment must all have access to the different experiments.

Using configuration management and product line software paradigms to support the experimentation process in software engineering.

There is no empirical evidence whatsoever to support most of the beliefs on which software construction is based. We do not yet know the adequacy, limits, qualities, costs and risks of the technologies used to develop software. Experimentation helps to check and convert beliefs and opinions into facts. This research is concerned with the replication area. Replication is a key component for gathering empirical evidence on software development that can be used in industry to build better software more efficiently. Replication has not been an easy thing to do in software engineering (SE) because the experimental paradigm applied to software development is still immature. Nowadays, a replication is executed mostly using a traditional replication package. But traditional replication packages do not appear, for some reason, to have been as effective as expected for transferring information among researchers in SE experimentation. The trouble spot appears to be the replication setup, caused by version management problems with materials, instruments, documents, etc. This has proved to be an obstacle to obtaining enough details about the experiment to be able to reproduce it as exactly as possible. We address the problem of information exchange among experimenters by developing a schema to characterize replications. We will adapt configuration management and product line ideas to support the experimentation process. This will enable researchers to make systematic decisions based on explicit knowledge rather than assumptions about replications. This research will output a replication support web environment. This environment will not only archive but also manage experimental materials flexibly enough to allow both similar and differentiated replications with massive experimental data storage. The platform should be accessible to several research groups working together on the same families of experiments.

RECLAMO: virtual and collaborative honeynets based on trust management and autonomous systems applied to intrusion management

Security intrusions in large systems is a problem due to its lack of scalability with the current IDS-based approaches. This paper describes the RECLAMO project, where an architecture for an Automated Intrusion Response System (AIRS) is being proposed. This system will infer the most appropriate response for a given attack, taking into account the attack type, context information, and the trust and reputation of the reporting IDSs. RECLAMO is proposing a novel approach: diverting the attack to a specific honeynet that has been dynamically built based on the attack information. Among all components forming the RECLAMO's architecture, this paper is mainly focused on defining a trust and reputation management model, essential to recognize if IDSs are exposing an honest behavior in order to accept their alerts as true. Experimental results confirm that our model helps to encourage or discourage the launch of the automatic reaction process.

Advanced system of planning and organization of classes at the university

This document is the result of a process of web development to create a tool that will allow to Cracow University of Technology consult, create and manage timetables. The technologies chosen for this purpose are Apache Tomcat Server, My SQL Community Server, JDBC driver, Java Servlets and JSPs for the server side. The client part counts on Javascript, jQuery, AJAX and CSS technologies to perform the dynamism. The document will justify the choice of these technologies and will explain some development tools that help in the integration and development of all this elements: specifically, NetBeans IDE and MySQL workbench have been used as helpful tools. After explaining all the elements involved in the development of the web application, the architecture and the code developed are explained through UML diagrams. Some implementation details related to security are also deeper explained through sequence diagrams. As the source code of the application is provided, an installation manual has been developed to run the project. In addition, as the platform is intended to be a beta that will be grown, some unimplemented ideas for future development are also exposed. Finally, some annexes with important files and scripts related to the initiation of the platform are attached. This project started through an existing tool that needed to be expanded. The main purpose of the project along its development has focused on setting the roots for a whole new platform that will replace the existing one. For this goal, it has been needed to make a deep inspection on the existing web technologies: a web server and a SQL database had to be chosen. Although the alternatives were a lot, Java technology for the server was finally selected because of the big community backwards, the easiness of modelling the language through UML diagrams and the fact of being free license software. Apache Tomcat is the open source server that can use Java Servlet and JSP technology. Related to the SQL database, MySQL Community Server is the most popular open-source SQL Server, with a big community after and quite a lot of tools to manage the server. JDBC is the driver needed to put in contact Java and MySQL. Once we chose the technologies that would be part of the platform, the development process started. After a detailed explanation of the development environment installation, we used UML use case diagrams to set the main tasks of the platform; UML class diagrams served to establish the existing relations between the classes generated; the architecture of the platform was represented through UML deployment diagrams; and Enhanced entity–relationship (EER) model were used to define the tables of the database and their relationships. Apart from the previous diagrams, some implementation issues were explained to make a better understanding of the developed code - UML sequence diagrams helped to explain this. Once the whole platform was properly defined and developed, the performance of the application has been shown: it has been proved that with the current state of the code, the platform covers the use cases that were set as the main target. Nevertheless, some requisites needed for the proper working of the platform have been specified. As the project is aimed to be grown, some ideas that could not be added to this beta have been explained in order not to be missed for future development. Finally, some annexes containing important configuration issues for the platform have been added after proper explanation, as well as an installation guide that will let a new developer get the project ready. In addition to this document some other files related to the project are provided: - Javadoc. The Javadoc containing the information of every Java class created is necessary for a better understanding of the source code. - database_model.mwb. This file contains the model of the database for MySQL Workbench. This model allows, among other things, generate the MySQL script for the creation of the tables. - ScheduleManager.war. The WAR file that will allow loading the developed application into Tomcat Server without using NetBeans. - ScheduleManager.zip. The source code exported from NetBeans project containing all Java packages, JSPs, Javascript files and CSS files that are part of the platform. - config.properties. The configuration file to properly get the names and credentials to use the database, also explained in Annex II. Example of config.properties file. - db_init_script.sql. The SQL query to initiate the database explained in Annex III. SQL statements for MySQL initialization. RESUMEN. Este proyecto tiene como punto de partida la necesidad de evolución de una herramienta web existente. El propósito principal del proyecto durante su desarrollo se ha centrado en establecer las bases de una completamente nueva plataforma que reemplazará a la existente. Para lograr esto, ha sido necesario realizar una profunda inspección en las tecnologías web existentes: un servidor web y una base de datos SQL debían ser elegidos. Aunque existen muchas alternativas, la tecnología Java ha resultado ser elegida debido a la gran comunidad de desarrolladores que tiene detrás, además de la facilidad que proporciona este lenguaje a la hora de modelarlo usando diagramas UML. Tampoco hay que olvidar que es una tecnología de uso libre de licencia. Apache Tomcat es el servidor de código libre que permite emplear Java Servlets y JSPs para hacer uso de la tecnología de Java. Respecto a la base de datos SQL, el servidor más popular de código libre es MySQL, y cuenta también con una gran comunidad detrás y buenas herramientas de modelado, creación y gestión de la bases de datos. JDBC es el driver que va a permitir comunicar las aplicaciones Java con MySQL. Tras elegir las tecnologías que formarían parte de esta nueva plataforma, el proceso de desarrollo tiene comienzo. Tras una extensa explicación de la instalación del entorno de desarrollo, se han usado diagramas de caso de UML para establecer cuáles son los objetivos principales de la plataforma; los diagramas de clases nos permiten realizar una organización del código java desarrollado de modo que sean fácilmente entendibles las relaciones entre las diferentes clases. La arquitectura de la plataforma queda definida a través de diagramas de despliegue. Por último, diagramas EER van a definir las relaciones entre las tablas creadas en la base de datos. Aparte de estos diagramas, algunos detalles de implementación se van a justificar para tener una mejor comprensión del código desarrollado. Diagramas de secuencia ayudarán en estas explicaciones. Una vez que toda la plataforma haya quedad debidamente definida y desarrollada, se va a realizar una demostración de la misma: se demostrará cómo los objetivos generales han sido alcanzados con el desarrollo actual del proyecto. No obstante, algunos requisitos han sido aclarados para que la plataforma trabaje adecuadamente. Como la intención del proyecto es crecer (no es una versión final), algunas ideas que se han podido llevar acabo han quedado descritas de manera que no se pierdan. Por último, algunos anexos que contienen información importante acerca de la plataforma se han añadido tras la correspondiente explicación de su utilidad, así como una guía de instalación que va a permitir a un nuevo desarrollador tener el proyecto preparado. Junto a este documento, ficheros conteniendo el proyecto desarrollado quedan adjuntos. Estos ficheros son: - Documentación Javadoc. Contiene la información de las clases Java que han sido creadas. - database_model.mwb. Este fichero contiene el modelo de la base de datos para MySQL Workbench. Esto permite, entre otras cosas, generar el script de iniciación de la base de datos para la creación de las tablas. - ScheduleManager.war. El fichero WAR que permite desplegar la plataforma en un servidor Apache Tomcat. - ScheduleManager.zip. El código fuente exportado directamente del proyecto de Netbeans. Contiene todos los paquetes de Java generados, ficheros JSPs, Javascript y CSS que forman parte de la plataforma. - config.properties. Ejemplo del fichero de configuración que permite obtener los nombres de la base de datos - db_init_script.sql. Las consultas SQL necesarias para la creación de la base de datos.

People, management and social responsability. The team: the main asset of any management

People, management and social responsability. The team: the main asset of any management. En colaboración con Dña. Montserrat Castellanos Moreno. Profesora Asociada de la Universidad Europea. Journal of Management and Marketing. Thomson Reuters. Zilina. Slovakia

Energy Management and Optimization for Video Decoders based on Functional-oriented Reconfiguration

In recent years, the increasing sophistication of embedded multimedia systems and wireless communication technologies has promoted a widespread utilization of video streaming applications. It has been reported in 2013 that youngsters, aged between 13 and 24, spend around 16.7 hours a week watching online video through social media, business websites, and video streaming sites. Video applications have already been blended into people daily life. Traditionally, video streaming research has focused on performance improvement, namely throughput increase and response time reduction. However, most mobile devices are battery-powered, a technology that grows at a much slower pace than either multimedia or hardware developments. Since battery developments cannot satisfy expanding power demand of mobile devices, research interests on video applications technology has attracted more attention to achieve energy-efficient designs. How to efficiently use the limited battery energy budget becomes a major research challenge. In addition, next generation video standards impel to diversification and personalization. Therefore, it is desirable to have mechanisms to implement energy optimizations with greater flexibility and scalability. In this context, the main goal of this dissertation is to find an energy management and optimization mechanism to reduce the energy consumption of video decoders based on the idea of functional-oriented reconfiguration. System battery life is prolonged as the result of a trade-off between energy consumption and video quality. Functional-oriented reconfiguration takes advantage of the similarities among standards to build video decoders reconnecting existing functional units. If a feedback channel from the decoder to the encoder is available, the former can signal the latter changes in either the encoding parameters or the encoding algorithms for energy-saving adaption. The proposed energy optimization and management mechanism is carried out at the decoder end. This mechanism consists of an energy-aware manager, implemented as an additional block of the reconfiguration engine, an energy estimator, integrated into the decoder, and, if available, a feedback channel connected to the encoder end. The energy-aware manager checks the battery level, selects the new decoder description and signals to build a new decoder to the reconfiguration engine. It is worth noting that the analysis of the energy consumption is fundamental for the success of the energy management and optimization mechanism. In this thesis, an energy estimation method driven by platform event monitoring is proposed. In addition, an event filter is suggested to automate the selection of the most appropriate events that affect the energy consumption. At last, a detailed study on the influence of the training data on the model accuracy is presented. The modeling methodology of the energy estimator has been evaluated on different underlying platforms, single-core and multi-core, with different characteristics of workload. All the results show a good accuracy and low on-line computation overhead. The required modifications on the reconfiguration engine to implement the energy-aware manager have been assessed under different scenarios. The results indicate a possibility to lengthen the battery lifetime of the system in two different use-cases.

Balancing management and preservation of mediterranean scattered oak woodlands (Dehesas) in human-dominated landscapes

Los sistemas agroforestales mediterráneos tienen una gran importancia ecológica y socioeconómica, y mantienen altos valores medioambientales y de diversidad biológica a la vez que producen importantes servicios ecosistémicos. Estos sistemas han sido testigos de diversos cambios rápidos y drásticos en su gestión y aprovechamiento durante el último siglo. La mayor parte de la investigación desarrollada en esta tesis doctoral ha sido llevada a cabo en las dehesas españolas. Esta tesis nos muestra: i) la evidencia de la existencia de un cambio global del estrato arbóreo y del manejo del pastoreo en el todo el área de distribución de la dehesa durante los últimos 60 años; ii) la importancia del papel que juega el arbolado disperso y el adecuado manejo del ganado en la mejora de la producción, calidad y diversidad de las comunidades herbáceas, que a su vez, un pasto herbáceo bien desarrollado es importante para la rentabilidad del sistema, evaluando estos efectos bajo distintos escenarios de clima y calidad de estación; y iii) la evidencia de la falta de regeneración en sistemas agroforestales mediterráneos bajo distintos tipos de manejo del pastoreo, y además se evalúa el crecimiento y desarrollo de las pocas plántulas existentes que serán las que aseguren la viabilidad y persistencia y de estos sistemas. El arbolado disperso de estos sistemas ha experimentado una reducción importante en su densidad arbórea y fracción de cabida cubierta durante el periodo entre 1950-1980 donde tuvieron lugar importantes transformaciones en la actividad agropecuaria. La cabaña ganadera de ovino disminuyó drásticamente en los años 70 en comparación a la de bovino que desde entonces ha aumentado progresivamente hasta la actualidad. Por otro lado, el mismo tipo de manejo del ganado doméstico (especialmente bovino) durante bastante tiempo (mínimo 30 años) provocó una reducción significativa de la densidad de las plántulas. Además la probabilidad de ocurrencia y la intensidad de daños por herbivoría fue mayor bajo pastoreo bovino (con daños más intensos y consistentes) que bajo pastoreo ovino o sin pastoreo doméstico (presencia de ciervos). También el patrón de crecimiento de las plantas jóvenes estuvo afectado por el tipo de manejo, generando plántulas achaparradas en el caso del bovino y plántulas esbeltas favoreciendo el crecimiento en altura en el caso del ovino. La presencia de un arbolado disperso generó una mayor diversidad y variación en la producción de las comunidades herbáceas según las condiciones de disponibilidad de agua. Especialmente, el ecotono como microhábitat sostuvo altos valores de diversidad herbácea. La presencia del ganado bajo pastoreo continuo de intensidad moderada a alta, especialmente el bovino, incrementó los rendimientos de producción y diversidad del estrato herbáceo. Los resultados de esta tesis nos muestran la importancia que tiene la existencia de un equilibrio entre la producción y la conservación de los sistemas agroforestales mediterráneos para obtener una producción sostenible de servicios ecosistémicos mientras se asegura la perpetuación del sistema a largo plazo. Es crucial diseñar planes de gestión incorporando objetivos de conservación que integren técnicas silvopastorales apropiadas para poder aplicar en los sistemas agroforestales mediterráneos. ABSTRACT Mediterranean scattered oak woodlands have great ecological and socio-economic importance, supporting high environmental and amenity values, and relatively rich biological diversity while producing important ecosystem services. They have been witnesses of different and fast changes developed in the last century. Most of the research developed in this dissertation has conducted within dehesas. This thesis provides: i) the global change evidence of the tree layer and grazing management experienced in the land-use range of a Mediterranean scattered oak woodland (dehesa) over the last 60 years; ii) the important role of scattered trees and adequate management grazing in the improvement of grassland yield, quality and diversity - which it is important, in turn, for the system profitability - under different climate scenarios and site quality; and iii) the lack of oak regeneration evidence under some given representative management regimes and how is the growth development of these plants to assure the viability and persistence of Mediterranean scattered oak woodlands. Tree layer experienced a significant reduction in dehesas during 1950-1980 period where the highest human impacts took place. Sheep herd decreased drastically during the 1970s and, in contrast, cattle have been increasing gradually since then. On the other hand, same livestock grazing management (especially cattle) during long time (minimum 30 years) within Mediterranean scattered oak woodlands reduced strongly the density of young oak plants and showed high probability of herbivory occurrence and intensity. Young plant growth pattern was greatly modified by livestock. Cattle grazing generated stunted plants and sheep grazing generated slender plants favoring the height growth. Microsites created by large trees modified the herbaceous yield according the water availability of the year and generated high plant diversity within herbaceous communities. Especially, ecotone microsite supported high values of herbaceous diversity. The presence of livestock species increased the herbaceous yield and maintained a more diverse community under continuous grazing at both moderate and high intensities; especially cattle. Thus, around the influence of scattered trees there is a high amount of different interactions among livestock, trees and grasslands maintaining and enhancing the quality of whole dehesa system. The results of this thesis highlight how important is balancing management and preservation of Mediterranean scattered oak woodlands to obtain the optimum ecosystem services while the system conservation is assured for a long-term. It is crucial to design management plans with conservation goals that include appropriate silvopastoral practices in Mediterranean scattered oak woodlands.

Balancing management and preservation of mediterranean scattered oak woodlands (Dehesas) in human-dominated landscapes

Los sistemas agroforestales mediterráneos tienen una gran importancia ecológica y socioeconómica, y mantienen altos valores medioambientales y de diversidad biológica a la vez que producen importantes servicios ecosistémicos. Estos sistemas han sido testigos de diversos cambios rápidos y drásticos en su gestión y aprovechamiento durante el último siglo. La mayor parte de la investigación desarrollada en esta tesis doctoral ha sido llevada a cabo en las dehesas españolas. Esta tesis nos muestra: i) la evidencia de la existencia de un cambio global del estrato arbóreo y del manejo del pastoreo en el todo el área de distribución de la dehesa durante los últimos 60 años; ii) la importancia del papel que juega el arbolado disperso y el adecuado manejo del ganado en la mejora de la producción, calidad y diversidad de las comunidades herbáceas, que a su vez, un pasto herbáceo bien desarrollado es importante para la rentabilidad del sistema, evaluando estos efectos bajo distintos escenarios de clima y calidad de estación; y iii) la evidencia de la falta de regeneración en sistemas agroforestales mediterráneos bajo distintos tipos de manejo del pastoreo, y además se evalúa el crecimiento y desarrollo de las pocas plántulas existentes que serán las que aseguren la viabilidad y persistencia y de estos sistemas. El arbolado disperso de estos sistemas ha experimentado una reducción importante en su densidad arbórea y fracción de cabida cubierta durante el periodo entre 1950-1980 donde tuvieron lugar importantes transformaciones en la actividad agropecuaria. La cabaña ganadera de ovino disminuyó drásticamente en los años 70 en comparación a la de bovino que desde entonces ha aumentado progresivamente hasta la actualidad. Por otro lado, el mismo tipo de manejo del ganado doméstico (especialmente bovino) durante bastante tiempo (mínimo 30 años) provocó una reducción significativa de la densidad de las plántulas. Además la probabilidad de ocurrencia y la intensidad de daños por herbivoría fue mayor bajo pastoreo bovino (con daños más intensos y consistentes) que bajo pastoreo ovino o sin pastoreo doméstico (presencia de ciervos). También el patrón de crecimiento de las plantas jóvenes estuvo afectado por el tipo de manejo, generando plántulas achaparradas en el caso del bovino y plántulas esbeltas favoreciendo el crecimiento en altura en el caso del ovino. La presencia de un arbolado disperso generó una mayor diversidad y variación en la producción de las comunidades herbáceas según las condiciones de disponibilidad de agua. Especialmente, el ecotono como microhábitat sostuvo altos valores de diversidad herbácea. La presencia del ganado bajo pastoreo continuo de intensidad moderada a alta, especialmente el bovino, incrementó los rendimientos de producción y diversidad del estrato herbáceo. Los resultados de esta tesis nos muestran la importancia que tiene la existencia de un equilibrio entre la producción y la conservación de los sistemas agroforestales mediterráneos para obtener una producción sostenible de servicios ecosistémicos mientras se asegura la perpetuación del sistema a largo plazo. Es crucial diseñar planes de gestión incorporando objetivos de conservación que integren técnicas silvopastorales apropiadas para poder aplicar en los sistemas agroforestales mediterráneos. ABSTRACT Mediterranean scattered oak woodlands have great ecological and socio-economic importance, supporting high environmental and amenity values, and relatively rich biological diversity while producing important ecosystem services. They have been witnesses of different and fast changes developed in the last century. Most of the research developed in this dissertation has conducted within dehesas. This thesis provides: i) the global change evidence of the tree layer and grazing management experienced in the land-use range of a Mediterranean scattered oak woodland (dehesa) over the last 60 years; ii) the important role of scattered trees and adequate management grazing in the improvement of grassland yield, quality and diversity - which it is important, in turn, for the system profitability - under different climate scenarios and site quality; and iii) the lack of oak regeneration evidence under some given representative management regimes and how is the growth development of these plants to assure the viability and persistence of Mediterranean scattered oak woodlands. Tree layer experienced a significant reduction in dehesas during 1950-1980 period where the highest human impacts took place. Sheep herd decreased drastically during the 1970s and, in contrast, cattle have been increasing gradually since then. On the other hand, same livestock grazing management (especially cattle) during long time (minimum 30 years) within Mediterranean scattered oak woodlands reduced strongly the density of young oak plants and showed high probability of herbivory occurrence and intensity. Young plant growth pattern was greatly modified by livestock. Cattle grazing generated stunted plants and sheep grazing generated slender plants favoring the height growth. Microsites created by large trees modified the herbaceous yield according the water availability of the year and generated high plant diversity within herbaceous communities. Especially, ecotone microsite supported high values of herbaceous diversity. The presence of livestock species increased the herbaceous yield and maintained a more diverse community under continuous grazing at both moderate and high intensities; especially cattle. Thus, around the influence of scattered trees there is a high amount of different interactions among livestock, trees and grasslands maintaining and enhancing the quality of whole dehesa system. The results of this thesis highlight how important is balancing management and preservation of Mediterranean scattered oak woodlands to obtain the optimum ecosystem services while the system conservation is assured for a long-term. It is crucial to design management plans with conservation goals that include appropriate silvopastoral practices in Mediterranean scattered oak woodlands.

A web-based application for the management and evaluation of tutoring requests in PBL-based massive laboratories

One important steps in a successful project-based-learning methodology (PBL) is the process of providing the students with a convenient feedback that allows them to keep on developing their projects or to improve them. However, this task is more difficult in massive courses, especially when the project deadline is close. Besides, the continuous evaluation methodology makes necessary to find ways to objectively and continuously measure students' performance without increasing excessively instructors' work load. In order to alleviate these problems, we have developed a web service that allows students to request personal tutoring assistance during the laboratory sessions by specifying the kind of problem they have and the person who could help them to solve it. This service provides tools for the staff to manage the laboratory, for performing continuous evaluation for all students and for the student collaborators, and to prioritize tutoring according to the progress of the student's project. Additionally, the application provides objective metrics which can be used at the end of the subject during the evaluation process in order to support some students' final scores. Different usability statistics and the results of a subjective evaluation with more than 330 students confirm the success of the proposed application.

The structure and organization within the membrane of the helices composing the pore-forming domain of Bacillus thuringiensis δ-endotoxin are consistent with an “umbrella-like” structure of the pore

The aim of this study was to elucidate the mechanism of membrane insertion and the structural organization of pores formed by Bacillus thuringiensis δ-endotoxin. We determined the relative affinities for membranes of peptides corresponding to the seven helices that compose the toxin pore-forming domain, their modes of membrane interaction, their structures within membranes, and their orientations relative to the membrane normal. In addition, we used resonance energy transfer measurements of all possible combinatorial pairs of membrane-bound helices to map the network of interactions between helices in their membrane-bound state. The interaction of the helices with the bilayer membrane was also probed by a Monte Carlo simulation protocol to determine lowest-energy orientations. Our results are consistent with a situation in which helices α4 and α5 insert into the membrane as a helical hairpin in an antiparallel manner, while the other helices lie on the membrane surface like the ribs of an umbrella (the “umbrella model”). Our results also support the suggestion that α7 may serve as a binding sensor to initiate the structural rearrangement of the pore-forming domain.

Localization of the C terminus of the assembly domain of hepatitis B virus capsid protein: Implications for morphogenesis and organization of encapsidated RNA

The capsid protein of hepatitis B virus, consisting of an “assembly” domain (residues 1–149) and an RNA-binding “protamine” domain (residues 150–183), assembles from dimers into icosahedral capsids of two different sizes. The C terminus of the assembly domain (residues 140–149) functions as a morphogenetic switch, longer C termini favoring a higher proportion of the larger capsids, it also connects the protamine domain to the capsid shell. We now have defined the location of this peptide in capsids assembled in vitro by engineering a mutant assembly domain with a single cysteine at its C terminus (residue 150), labeling it with a gold cluster and visualizing the cluster by cryo-electron microscopy. The labeled protein is unimpaired in its ability to form capsids. Our density map reveals a single undecagold cluster under each fivefold and quasi-sixfold vertex, connected to sites at either end of the undersides of the dimers. Considering the geometry of the vertices, the C termini must be more crowded at the fivefolds. Thus, a bulky C terminus would be expected to favor formation of the larger (T = 4) capsids, which have a greater proportion of quasi-sixfolds. Capsids assembled by expressing the full-length protein in Escherichia coli package bacterial RNAs in amounts equivalent to the viral pregenome. Our density map of these capsids reveals a distinct inner shell of density—the RNA. The RNA is connected to the protein shell via the C-terminal linkers and also makes contact around the dimer axes.

Numbers and Organization of RNA Polymerases, Nascent Transcripts, and Transcription Units in HeLa Nuclei

Using HeLa cells, we have developed methods to determine 1) the number of RNA polymerases that are active at any moment, 2) the number of transcription sites, and 3) the number of polymerases associated with one transcription unit. To count engaged polymerases, cells were encapsulated in agarose, permeabilized, treated with ribonuclease, and the now-truncated transcripts extended in [32P]uridine triphosphate; then, the number of growing transcripts was calculated from the total number of nucleotides incorporated and the average increment in length of the transcripts. Approximately 15,000 transcripts were elongated by polymerase I, and ∼75,000 were elongated by polymerases II and III. Transcription sites were detected after the cells were grown in bromouridine for <2.5 min, after which the resulting bromo-RNA was labeled with gold particles; electron microscopy showed that most extranucleolar transcripts were concentrated in ∼2400 sites with diameters of ∼80 nm. The number of polymerases associated with a transcription unit was counted after templates were spread over a large area; most extranucleolar units were associated with one elongating complex. These results suggest that many templates are attached in a “cloud” of loops around a site; each site, or transcription “factory,” would contain ∼30 active polymerases and associated transcripts.

The neural development and organization of letter recognition: Evidence from functional neuroimaging, computational modeling, and behavioral studies

Although much of the brain’s functional organization is genetically predetermined, it appears that some noninnate functions can come to depend on dedicated and segregated neural tissue. In this paper, we describe a series of experiments that have investigated the neural development and organization of one such noninnate function: letter recognition. Functional neuroimaging demonstrates that letter and digit recognition depend on different neural substrates in some literate adults. How could the processing of two stimulus categories that are distinguished solely by cultural conventions become segregated in the brain? One possibility is that correlation-based learning in the brain leads to a spatial organization in cortex that reflects the temporal and spatial clustering of letters with letters in the environment. Simulations confirm that environmental co-occurrence does indeed lead to spatial localization in a neural network that uses correlation-based learning. Furthermore, behavioral studies confirm one critical prediction of this co-occurrence hypothesis, namely, that subjects exposed to a visual environment in which letters and digits occur together rather than separately (postal workers who process letters and digits together in Canadian postal codes) do indeed show less behavioral evidence for segregated letter and digit processing.