Espacio web para el diseño de un museo virtual de la historia de la informática

El objetivo principal de crear un espacio web para el Museo Histórico de la Informática (MHI) perteneciente a la Escuela Técnica Superior de Ingenieros Informáticos (ETSIINF) de la Universidad Politécnica de Madrid (UPM) es la difusión de la historia de la informática entre el público en general. Si bien es cierto que existe ya una página web de consulta del MHI con algunas imágenes y contenido sobre los objetos que allí se exhiben, es también reseñable que se trata de un espacio obsoleto, lleno de carencias y extremadamente difícil de gestionar y actualizar, por lo que se hacía imprescindible actualizar el diseño del espacio web, los contenidos y el sistema de gestión de los mismos, cosa que es de gran interés para un lugar divulgativo. En la actualidad, existen maneras mucho más amigables para el usuario de navegar por una web; y de la misma manera para un administrador, gestionar el contenido de la misma y mantener a los usuarios bien informados de todo lo que se ofrece en cada momento. Esto es posible gracias a los sistemas de gestión de contenidos o content management system (CMS), de los que se hablará lo largo de todo el documento. Estos sistemas, dan una facilidad mucho mayor a los encargados de llevar al día una página web, sin tener que saber de programación, lenguajes o informática en general, ya que incorporan paneles de control muy intuitivos y fáciles de manejar, que son una ventaja tanto para ellos como para los usuarios. Es por esta razón que, hay páginas web como la de las empresas IKEA, Ubuntu o, en especial para el caso que compete al documento, el museo del Louvre usan gestores de contenidos para sus páginas web. Y es que las ventajas y facilidades que ofrece un CMS son realmente interesantes y se tratará de todas ellas en el documento, de la elección del CMS que mejor se ajusta a los requisitos del museo, las restricciones a la hora del despliegue en el ámbito de la ETSIINF y de cómo mejorará esto la calidad visual y divulgativa del MHI. Este trabajo se desarrolla a lo largo de 11 capítulos, en los que se muestra como construir un sitio web, las posibilidades y la elección final para este caso. En el primer capítulo se hace una pequeña introducción de lo que es el proyecto, se especifican los objetivos, la motivación del mismo y el alcance que tiene. En el segundo capítulo se muestra la información que se ha recopilado en el trabajo de investigación que se hace previo al desarrollo. En él se muestran los distintos tipos de páginas web, que tecnologías y lenguajes se pueden usar para su construcción, una comparativa sobre otras entidades similares al MHI, las limitaciones que presenta el entorno y la elección final que se consideró más adecuada para este caso. En el capítulo tres se empieza a desarrollar la solución a través del diseño. Aquí se puede encontrar el diseño de más bajo nivel que se le presenta al cliente para sentar las bases del trabajo, el diseño de alto nivel con un mayor grado de realidad que el anterior y una preparación de lo que serán los planes de prueba. El capítulo cuatro muestra todo lo que se ha usado en la implementación y la integración de la página web: herramientas, tecnologías, plantillas de diseño y módulos que proporcionan distintas funcionalidades. Llegados al capítulo cinco, se puede ver una detallada documentación de los resultados de las pruebas de usabilidad y accesibilidad realizadas, y las conclusiones que subyacen de estas. Una vez acabada la implementación del sitio web del MHI, en el capítulo seis se intenta hacer una labor de consultoría, mostrando precios y presupuestos de las distintas tareas que se han llevado a cabo para la realización de este proyecto. En el capítulo siete se extraen las conclusiones de todo lo acontecido en los capítulos anteriores y en el ocho unas líneas sobre posibles trabajos futuros que se podrían llevar a cabo en base a lo que ya se tiene en la institución, incluido este trabajo. Para facilitar la comprensión y satisfacer la curiosidad del lector, se ha incluido en el capítulo nueve la bibliografía con toda la documentación consultada y en el diez un glosario de términos para la aclaración de términos y acrónimos más técnicos. Para acabar, en el capítulo once se anexionan tanto el documento que se usó para las pruebas de usabilidad como un manual de administrador para el sitio web, que hace más amigable el entorno para las personas que lo tengan que mantener en un futuro.---ABSTRACT---The main goal of creating a website for the Informatics Historical Museum (“Museo Histórico de la Informática” or MHI) located in the Higher Technical School of Informatics Engineers (“Escuela Técnica Superior de Ingenieros Informáticos” or ETSIINF) of the Polytechnic University of Madrid (“Universidad Politécnica de Madrid” UPM), is to promote the aforementioned museum as to increase its reach to the public. While it is true that there already is a consulting website with some pictures and information about the items which are displayed in the building, it is outdated and the data is scarce. Moreover, it is extremely complicated to manage and to regularly update the web page, which is very important for informative/broadcasting media. Currently, there are easier ways for the users of a website to consult whatever information they want, as well as it is now easy for a website manager to display new content and to keep the users informed about what is been offered at every moment. This is possible because of content management systems (CMS), which will be discussed throughout the entire paper. These systems make it easier to use for the administrator of a website to keep it up to date without the necessity for them of having any knowledge or skills in programming, languages or computing, because the systems have an intuitive control panel that is easy to use, which is an advantage for both managers and users. Because of all these reasons, there are lots of companies that use this kind of systems, such as IKEA, Ubuntu or, especially, the Louvre Museum, to which we direct our attention all throughout this report. It is easy to notice that these systems have an important and very interesting number of perks and benefits. In the next chapters of the document we will explain the benefits of the program as well as the choice on the kind of CMS that best suits the requirements of the museum and, finally, the restrictions of the school for the deployment and of how all of this will improve the visual and informational qualities of the MHI. This work is developed over 11 chapters, shown how to build up a website, the possibilities and the final choice for this case. In the first chapter a brief introduction of the project, goals, motivation and scope thereof having specified are done. Before the development of the website, the second chapter shows de information of the researching work. It discusses the different types of websites, technologies and languages that can be used for build-up, a comparison of similar entities to MHI, the limitations of the environment and the final choice was considered more appropriate for this case. Chapter three begins to develop the design of the solution. Here there are the lowest level design that presents the customer to fix any problem, the high level design with a higher degree of reality than the last and the test plans. Chapter four shows everything that has been used in the implementation and integration of the website: tools, technologies, design templates and modules that provide different functionalities. Reaching the fifth chapter, you can see a detailed documentation of the results of the usability and accessibility tests made to some users and the conclusions of it. Once the implementation of MHI website is done, in chapter six there is a consultancy work, showing prices and budgets of the different tasks which were carried out for this project. In chapter seven there are the conclusions of what happened in the previous chapters and eight chapter shows possible future works that could be carried out based on what the institution already have, including this work. To make easier to the reader understand this paper and satisfy his curiosity, the chapter nine includes the bibliography consulted with all the documentation and chapter ten has a glossary of terms and an explanation of technical terms and acronyms. Finally, in chapter eleven there are attached both the document that was used for usability testing as a manual administrator for the website, making the environment friendlier for people who have to maintain it in the future.

Herramienta de partes de actividad

Para entender el proyecto de una manera muy simplificada lo podríamos resumir de la siguiente forma: Se trata de una aplicación que permite a los usuarios imputar las horas de las tareas que realizan de una forma sencilla, y que los gestores de estos usuarios, mediante informes ejecutivos, puedan analizar las dedicaciones y costes de los proyectos que éstos gestionan. Además se dispondrá de pantallas para administrar todas las entidades que están implicadas en las imputaciones. Las imputaciones se realizarán agrupándolas por el concepto “Parte”. Los Partes dividen los meses en grupos de 10 días, así cada usuario tendrá que imputar las horas que realiza cada 10 días, enviando a su supervisor el parte para que este pueda revisarlo y analizar los resultados de las imputaciones a través de los informes. Vamos a resumir las entidades que intervienen en la aplicación y sus relaciones de la siguiente forma: - Los usuarios tendrán asignados una serie de cargos, los cuales definirán las posibles tareas que podrán realizar categorizadas en los conceptos Proceso y Subproceso. Un ejemplo de esta relación podría ser el cargo “Programador” podrá realizar tareas de tipo Proceso “Desarrollo” y tipos de Subproceso “Programación” y “Pruebas unitarias” - También se asignarán a los usuarios los proyectos en los que trabajan. Estos proyectos pertenecerán a una unidad (se podría equiparar el concepto unidad por ejemplo a clientes, departamentos, etc). - También se podrán asignar a los usuarios las peticiones concretas de proyectos en los que intervienen. Una petición podría ser una incidencia detectada en un proyecto, una mejora concreta en un proyecto de mantenimiento, un soporte realizado para arreglar unos datos incorrectos, etc. - A los usuarios se le asignarán unos costes que podrán ser distintos por periodos de tiempo con el fin de conocer el coste exacto de una hora realizada en la fecha en la que fue realizada. - A los usuarios se les asignará un Perfil de usuario. Este perfil de usuario definirá los permisos sobre los módulos disponibles de la herramienta. Habrá un permiso por cada entidad, dividiéndolos en Alta, Baja, Modificación y Consulta. Además de estos permisos habrá permisos especiales sobre módulos que no se refieran al manteniendo de entidades pero que sí requieran de un permiso especial para poder ejecutarse como por ejemplo poder ejecutar los informes de costes, poder realizar y consultar partes de otros usuarios además de los propios del usuario, etc. Los perfiles serán dinámicos. Se podrán crear tantos perfiles como se requieran, cada uno con los permisos que se le asignen. - Los usuarios imputarán las horas que realizan categorizando por tarea los siguientes conceptos: Proyecto, Petición (opcional), Proceso, Subproceso, Descripción de la tarea y horas invertidas en los días de la decena. La aplicación permite poder adaptarse a las necesidades de diseño de la organización que la utilice y pueden crearse tantas instancias de la aplicación como se necesiten, compartiendo ono la base de datos entre varias de las instancias, sin necesidad de duplicar el código en el servidor, lo que facilita el mantenimiento de la aplicación cuando se añaden mejoras y se solucionan incidencias. También facilita el poder vender licencias de uso de la aplicación a otras entidades teniendo una sola versión de la aplicación en la nube, lo que hace muy bajo el coste de la venta de licencias aumentando así los beneficios por venta. ABSTRACT To understand the project in a very simplified way we could summarize as follows: Its an application that allows users to charge the hours of tasks that they realize in a easy way, and that the managers of these users, by executive reports, can analyze dedications and costs of projects that they manage. Furthermore screens will be available to manage all entities that are involved in the charge of hours. The charge of hours will be grouped by the concept "Timesheet". The Timesheets divided the months in groups of 10 days, so each user will have to charge the hours every 10 days by sending his supervisor the thimesheet so that it can review it and analyze the results of the charges of hours by reports.Let's summarize the entities involved in the application and their relationships as follows: - Users will be assigned to one or more positions, which define the possible tasks that can perform categorized into concepts Process and Subprocess. An example of this relationship could be the position "Developer" may do tasks of process type "Development" and subprocess types "Programming" and "Unit Testing" - Users will also be assigned to projects in which they work. These projects belongs to a unit (Some possible examples of unit could be customers, departments, ...). - You can also assign users to specific requests for projects thar they are involved. A request could be an issue detected in a project, a concrete improvement in a maintenance project, a support made to fix some incorrect data, ... - Users will be assigned to one or more costs that may be different for periods of time in order to know the exact cost of an hour on the date on which it was made. - Users will be assigned to a User Profile. This user profile define the available permissions on modules of tool. There will be a permission for each entity, dividing them into Insert, Delete, Modify and Read. In addition to these permissions will be special permissions on modules that are not related to maintaining entities but if require special permission to execute such as to execute the cost reports, to make and read Timesheets of others users in addition to themselve user, etc. The profiles will be dynamic. We can create many profiles as we required, each with the permissions that are assigned. - Users charge the hours that they work categorizing per task the following: Project, Request (optional), Process, Subprocess, task description and the hours that they used in the ten days period. The application allows to adapt to the needs of design of the organization that use the application and allows create many instances of the application as needed, yes or not sharing the database among multiple instances without duplicating the code on the server, which facilitates maintenance of the application when improvements are added and incidents are resolved. It also facilitates to sell licenses to use the application to other organizations having a single version of the application in the cloud, making it very low cost from the sale of licenses and increasing profits per sale.

O derramamento do Espírito : fortalecimento dos enfraquecidos

Esta tese se insere dentro da leitura bíblica latino-americana trazendo, nessa ótica, a riqueza e o desafio do que significa ler a Bíblia a partir de um contexto de opressão como é o nosso continente. Nos servimos do roteiro das ciências bíblicas e dos métodos exegéticos modernos. É uma pesquisa bibliográfica onde, eventualmente, consideramos o nível experiêncial, por exemplo quando trabalhamos o tema do derramamento do Espírito nos pentecostais chilenos. Esse capítulo, IV, segue o roteiro da hipótese central de nossa tese na medida que firmamos o princípio de que os pentecostais fazem parte do amplo e diverso grupo dos enfraquecidos como produto de um sistema excludente, baseado na exploração social. Numa situação de crise, o anúncio do derramamento do Espírito é um sinal de salvação, de perspectivas futuras, de conservação e prolongação da vida. Esta tese relê o tema do derramamento do Espírito, a partir de Jl 3,1-5. O tema é analisado num contexto social concreto, onde os enfraquecidos recebem o Espírito do Senhor. O eixo que nos permite fazer esta leitura encontra sua referencial nas pessoas setores sociais - mencionadas como beneficiadas diretas da ação do Espírito. São pessoas que representam setores diferentes; jovens, escravos e escravas, formam o grupo que sustenta e faz funcionar o sistema imperial persa grego - baseado na exploração e mão de obra barata. Eles, juntamente com os idosos que não produzem mais, são a base de sustentação da pirâmide social. Como demostraremos no decorrer da tese, nossa hipótese somente é possível se duas conjunturas se cruzam. Para isso, em num primeiro momento, localizamos e demostramos que o livro de Joel deve ser situado no contexto da literatura apocalíptica (nascimento). Em seguida, demonstramos que o livro de Joel, como produto literário final, deve ser localizado no contexto histórico do pós-exílio, isto é, no tempo dos impérios persa e grego. A confirmação destes dois aspectos desenvolvidos nos capítulos I e II, nos permitem, na análise literária, capítulo III, aprofundar, mediante a análise exegética, e confirmar a nossa hipótese central. Com estes três capítulos demostramos que o derramamento do Espírito do Senhor tem como destinatários preferenciais, senão exclusivos, os setores enfraquecidos pela política social, econômica e religiosa dos impérios persa e grego. Com estes três capítulos desenvolvidos, no quarto capítulo analisamos uma experiência concreta, de um setor majoritariamente pobre que tem se apropriado do texto de Jl 3,1-5, e encontrado nele uma alternativa social, política e religiosa para se manter fiel ao Senhor e por quase um século recria e revive a experiência do derramamento do Espírito. Os velhos, os jovens, os escravos e as escravas, formam o setor dos enfraquecidos. É o setor que nada espera, e que nada terá da parte dos impérios e que, como os pentecostais, pela sua situação de enfraquecimento, acredita que somente uma intervenção externa pode mudar o seu futuro, pode trazer de volta a esperança. Essa intervenção externa começa a ser possível pelo derramamento do Espírito e se concretiza na chegada do dia de Javé, que será grande e terrível, onde o sol e a lua se unem para indicar o monte de Sião e a cidade de Jerusalém como lugar de adoração, refúgio e salvação. Esta perspectiva da ação do Espírito é possível somente no período do pós-exílio. Nesse tempo o Espírito adquire uma dimensão ampla e inclusiva agindo sobre diversos setores sociais, independe de serem ou não judeus.(AU)

ANALISANDO A RELAÇÃO DOS VOLUMES DE EXPORTAÇÃO E DE IMPORTAÇÃO, PIB, TAXAS DE CÂMBIO E INFLAÇÃO NO PERÍODO DE 2004 A 2014.

Esta dissertação visa deslumbrar uma análise macroeconômica do Brasil, especialmente no que se refere à relação dos índices mensais dos volumes das exportações e das importações com os volumes mensais do PIB, da Taxa SELIC e as Taxas de Câmbio, conforme dados coletados no período de janeiro de 2004 a dezembro de 2014, através de pesquisa literária referente aos históricos sobre cada conceito envolvido no âmbito da macroeconomia das varáveis estudadas. Foi realizado um estudo de caso embasado em dados de sites governamentais, no período delimitado, empregando-se o método de regressão linear, com base na Teoria da correlação de Pearson, demonstrando os resultados obtidos no período do estudo para as varáveis estudadas. Desta maneira, conseguiu-se estudar e analisar como as variáveis dependentes (resposta): volume das exportações e volume das importações estão relacionadas com as varáveis independentes (explicativas): PIB, Taxa Selic e taxa de Câmbio. Os resultados apurados no presente estudo permitem identificar que existe correlação moderada e negativa, quando analisadas a Taxa Selic e a Taxa de Câmbio com os volumes das exportações e das importações, enquanto o PIB apresenta correlação forte e positiva na análise com os volumes das exportações e das importações

A factor IX-deficient mouse model for hemophilia B gene therapy

We have generated a mouse where the clotting factor IX (FIX) gene has been disrupted by homologous recombination. The FIX nullizygous (−/−) mouse was devoid of factor IX antigen in plasma. Consistent with the bleeding disorder, the factor IX coagulant activities for wild-type (+/+), heterozygous (+/−), and homozygous (−/−) mice were 92%, 53%, and <5%, respectively, in activated partial thromboplastin time assays. Plasma factor IX activity in the deficient mice (−/−) was restored by introducing wild-type murine FIX gene via adenoviral vectors. Thus, these factor IX-deficient mice provide a useful animal model for gene therapy studies of hemophilia B.

The anaphase inhibitor of Saccharomyces cerevisiae Pds1p is a target of the DNA damage checkpoint pathway

Inhibition of DNA replication and physical DNA damage induce checkpoint responses that arrest cell cycle progression at two different stages. In Saccharomyces cerevisiae, the execution of both checkpoint responses requires the Mec1 and Rad53 proteins. This observation led to the suggestion that these checkpoint responses are mediated through a common signal transduction pathway. However, because the checkpoint-induced arrests occur at different cell cycle stages, the downstream effectors mediating these arrests are likely to be distinct. We have previously shown that the S. cerevisiae protein Pds1p is an anaphase inhibitor and is essential for cell cycle arrest in mitosis in the presence DNA damage. Herein we show that DNA damage, but not inhibition of DNA replication, induces the phosphorylation of Pds1p. Analyses of Pds1p phosphorylation in different checkpoint mutants reveal that in the presence of DNA damage, Pds1p is phosphorylated in a Mec1p- and Rad9p-dependent but Rad53p-independent manner. Our data place Pds1p and Rad53p on parallel branches of the DNA damage checkpoint pathway. We suggest that Pds1p is a downstream target of the DNA damage checkpoint pathway and that it is involved in implementing the DNA damage checkpoint arrest specifically in mitosis.

The change in hydrogen bond strength accompanying charge rearrangement: Implications for enzymatic catalysis

The equilibrium for formation of the intramolecular hydrogen bond (KHB) in a series of substituted salicylate monoanions was investigated as a function of ΔpKa, the difference between the pKa values of the hydrogen bond donor and acceptor, in both water and dimethyl sulfoxide. The dependence of log KHB upon ΔpKa is linear in both solvents, but is steeper in dimethyl sulfoxide (slope = 0.73) than in water (slope = 0.05). Thus, hydrogen bond strength can undergo substantially larger increases in nonaqueous media than aqueous solutions as the charge density on the donor or acceptor atom increases. These results support a general mechanism for enzymatic catalysis, in which hydrogen bonding to a substrate is strengthened as charge rearranges in going from the ground state to the transition state; the strengthening of the hydrogen bond would be greater in a nonaqueous enzymatic active site than in water, thus providing a rate enhancement for an enzymatic reaction relative to the solution reaction. We suggest that binding energy of an enzyme is used to fix the substrate in the low-dielectric active site, where the strengthening of the hydrogen bond in the course of a reaction is increased.

Parametrizations of elliptic curves by Shimura curves and by classical modular curves

Fix an isogeny class

Escherichia coli Nth and human hNTH1 DNA glycosylases are involved in removal of 8-oxoguanine from 8-oxoguanine/guanine mispairs in DNA

The spectrum of DNA damage caused by reactive oxygen species includes a wide variety of modifications of purine and pyrimidine bases. Among these modified bases, 7,8-dihydro-8-oxoguanine (8-oxoG) is an important mutagenic lesion. Base excision repair is a critical mechanism for preventing mutations by removing the oxidative lesion from the DNA. That the spontaneous mutation frequency of the Escherichia coli mutT mutant is much higher than that of the mutM or mutY mutant indicates a significant potential for mutation due to 8-oxoG incorporation opposite A and G during DNA replication. In fact, the removal of A and G in such a situation by MutY protein would fix rather than prevent mutation. This suggests the need for differential removal of 8-oxoG when incorporated into DNA, versus being generated in situ. In this study we demonstrate that E.coli Nth protein (endonuclease III) has an 8-oxoG DNA glycosylase/AP lyase activity which removes 8-oxoG preferentially from 8-oxoG/G mispairs. The MutM and Nei proteins are also capable of removing 8-oxoG from mispairs. The frequency of spontaneous G:C→C:G transversions was significantly increased in E.coli CC103mutMnthnei mutants compared with wild-type, mutM, nth, nei, mutMnei, mutMnth and nthnei strains. From these results it is concluded that Nth protein, together with the MutM and Nei proteins, is involved in the repair of 8-oxoG when it is incorporated opposite G. Furthermore, we found that human hNTH1 protein, a homolog of E.coli Nth protein, has similar DNA glycosylase/AP lyase activity that removes 8-oxoG from 8-oxoG/G mispairs.

N2 fixation in marine heterotrophic bacteria: dynamics of environmental and molecular regulation.

Molecular and immunological techniques were used to examine N2 fixation in a ubiquitous heterotrophic marine bacterium, the facultative anaerobic Vibrio natriegens. When batch cultures were shifted from aerobic N-replete to anaerobic N-deplete conditions, transcriptional and post-translational regulation of N2 fixation was observed. Levels of nifHDK mRNA encoding the nitrogenase enzyme were highest at 140 min postshift and undetectable between 6 and 9 h later. Immunologically determined levels of nitrogenase enzyme (Fe protein) were highest between 6 and 15 h postshift, and nitrogenase activity peaked between 6 and 9 h postshift, declining by a factor of 2 after 12-15 h. Unlike their regulation in cyanobacteria, Fe protein and nitrogenase activity were present when nifHDK mRNA was absent in V. natriegens, indicating that nitrogenase is stored and stable under anaerobic conditions. Both nifHDK mRNA and Fe protein disappeared within 40 min after cultures were shifted from N2-fixing conditions (anaerobic, N-deplete) to non- N2-fixing conditions (aerobic, N-enriched) but reappeared when shifted to conditions favoring N2 fixation. Thus, unlike other N2-fixing heterotrophic bacteria, nitrogenase must be resynthesized after aerobic exposure in V. natriegens. Immunological detection based on immunoblot (Western) analysis and immunogold labeling correlated positively with nitrogenase activity; no localization of nitrogenase was observed. Because V. natriegens continues to fix N2 for many hours after anaerobic induction, this species may play an important role in providing "new" nitrogen in marine ecosystems.

Reconstitution of repair-gap UV mutagenesis with purified proteins from Escherichia coli: a role for DNA polymerases III and II.

Using a cell-free system for UV mutagenesis, we have previously demonstrated the existence of a mutagenic pathway associated with nucleotide-excision repair gaps. Here, we report that this pathway can be reconstituted by using six purified proteins: UvrA, UvrB, UvrC, DNA helicase II, DNA polymerase III core, and DNA ligase. This establishes the minimal requirements for repair-gap UV mutagenesis. DNA polymerase II could replace DNA polymerase III, although less effectively, whereas DNA polymerase I, the major repair polymerase, could not. DNA sequence analysis of mutations generated in the in vitro reaction revealed a spectrum typical of mutations targeted to UV lesions. These observations suggest that repair-gap UV mutagenesis is performed by DNA polymerase III, and to a lesser extent by DNA polymerase II, by filling-in of a rare class of excision gaps that contain UV lesions.

X-ray structure of clotting factor IXa: active site and module structure related to Xase activity and hemophilia B.

Hereditary deficiency of factor IXa (fIXa), a key enzyme in blood coagulation, causes hemophilia B, a severe X chromosome-linked bleeding disorder afflicting 1 in 30,000 males; clinical studies have identified nearly 500 deleterious variants. The x-ray structure of porcine fIXa described here shows the atomic origins of the disease, while the spatial distribution of mutation sites suggests a structural model for factor X activation by phospholipid-bound fIXa and cofactor VIIIa. The 3.0-A-resolution diffraction data clearly show the structures of the serine proteinase module and the two preceding epidermal growth factor (EGF)-like modules; the N-terminal Gla module is partially disordered. The catalytic module, with covalent inhibitor D-Phe-1I-Pro-2I-Arg-3I chloromethyl ketone, most closely resembles fXa but differs significantly at several positions. Particularly noteworthy is the strained conformation of Glu-388, a residue strictly conserved in known fIXa sequences but conserved as Gly among other trypsin-like serine proteinases. Flexibility apparent in electron density together with modeling studies suggests that this may cause incomplete active site formation, even after zymogen, and hence the low catalytic activity of fIXa. The principal axes of the oblong EGF-like domains define an angle of 110 degrees, stabilized by a strictly conserved and fIX-specific interdomain salt bridge. The disorder of the Gla module, whose hydrophobic helix is apparent in electron density, can be attributed to the absence of calcium in the crystals; we have modeled the Gla module in its calcium form by using prothrombin fragment 1. The arched module arrangement agrees with fluorescence energy transfer experiments. Most hemophilic mutation sites of surface fIX residues occur on the concave surface of the bent molecule and suggest a plausible model for the membrane-bound ternary fIXa-FVIIIa-fX complex structure: fIXa and an equivalently arranged fX arch across an underlying fVIIIa subdomain from opposite sides; the stabilizing fVIIIa interactions force the catalytic modules together, completing fIXa active site formation and catalytic enhancement.

A second nitrogenase in vegetative cells of a heterocyst-forming cyanobacterium.

In many filamentous cyanobacteria nitrogen fixation occurs in differentiated cells called heterocysts. Filamentous strains that do not form heterocysts may fix nitrogen in vegetative cells, primarily under anaerobic conditions. We describe here two functional Mo-dependent nitrogenases in a single organism, the cyanobacterium Anabaena variabilis. Using a lacZ reporter with a fluorescent beta-galactoside substrate for in situ localization of gene expression, we have shown that the two clusters of nif genes are expressed independently. One nitrogenase functions only in heterocysts under either aerobic or anaerobic growth conditions, whereas the second nitrogenase functions only under anaerobic conditions in vegetative cells and heterocysts. Differences between the two nif clusters suggest that the nitrogenase that is expressed in heterocysts is developmentally regulated while the other is regulated by environmental factors.

Chloroplast gene sequence data suggest a single origin of the predisposition for symbiotic nitrogen fixation in angiosperms.

Of the approximately 380 families of angiosperms, representatives of only 10 are known to form symbiotic associations with nitrogen-fixing bacteria in root nodules. The morphologically based classification schemes proposed by taxonomists suggest that many of these 10 families of plants are only distantly related, engendering the hypothesis that the capacity to fix nitrogen evolved independently several, if not many, times. This has in turn influenced attitudes toward the likelihood of transferring genes responsible for symbiotic nitrogen fixation to crop species lacking this ability. Phylogenetic analysis of DNA sequences for the chloroplast gene rbcL indicates, however, that representatives of all 10 families with nitrogen-fixing symbioses occur together, with several families lacking this association, in a single clade. This study therefore indicates that only one lineage of closely related taxa achieved the underlying genetic architecture necessary for symbiotic nitrogen fixation in root nodules.