La casa crecedera. El crecimiento programado de la vivienda con innovación europea y economía de medios latinoamericana

El concepto de casa crecedera, tal y como lo conocemos en la actualidad, se acuñó por primera vez en 1932 en el concurso Das Wachsende Haus organizado por Martin Wagner y Hans Poelzig dentro del marco de la Exposición Internacional Sonne, Luft und Haus für alle, promovida por la Oficina de Turismo de la ciudad de Berlín. En dicho concurso, se definía este tipo de vivienda como aquella célula básica o vivienda semilla que, dependiendo de las necesidades y posibilidades de los habitantes, podía crecer mediante otras estancias, conformando una vivienda completa en sí misma en cada fase de crecimiento. Numerosos arquitectos de primer orden, tales como Walter Gropius, Bruno Taut, Erich Mendelsohn o Hans Scharoun, participaron en este concurso, abriendo una nueva vía de exploración dentro de la vivienda flexible, la del crecimiento programado. A partir de ese momento, en Europa, y subsecuentemente en EEUU y otras regiones desarrolladas, se iniciaron numerosas investigaciones teóricas y prácticas en torno al fenómeno del crecimiento en la vivienda desde un enfoque vinculado a la innovación, tanto espacial como técnica. Por otro lado, aunque dentro del marco de la arquitectura popular de otros países, ya se ensayaban viviendas crecederas desde el siglo XVIII debido a que, por su tamaño, eran más asequibles dentro del mercado. Desde los años treinta, numerosos países en vías de desarrollo tuvieron que lidiar con migraciones masivas del campo a la ciudad, por lo que se construyeron grandes conjuntos habitacionales que, en numerosos casos, estaban conformados por viviendas crecederas. En todos ellos, la aproximación al crecimiento de la vivienda se daba desde una perspectiva diferente a la de los países desarrollados. Se primaba la economía de medios, el uso de sistemas constructivos de bajo costo y, en muchos casos, se fomentaba incluso la autoconstrucción guiada, frente a las construcciones prefabricadas ensambladas por técnicos especializados que se proponían, por ejemplo, en los casos europeos. Para realizar esta investigación, se recopiló información de estas y otras viviendas. A continuación, se identificaron distintas maneras de producir el crecimiento, atendiendo a su posición relativa respecto de la vivienda semilla, a las que se denominó mecanismos de ampliación, utilizados indistintamente sin tener en cuenta la ubicación geográfica de cada casa. La cuestión de porqué se prefiere un mecanismo en lugar de otro en un caso determinado, desencadenó el principal objetivo de esta Tesis: la elaboración de un sistema de análisis y diagnóstico de la vivienda crecedera que, de acuerdo a determinados parámetros, permitiera indicar cuál es la ampliación o sucesión de ampliaciones óptimas para una familia concreta, en una ubicación establecida. Se partió de la idea de que el crecimiento de la vivienda está estrechamente ligado a la evolución de la unidad de convivencia que reside en ella, de manera que la casa se transformó en un hábitat dinámico. Además se atendió a la complejidad y variabilidad del fenómeno, sujeto a numerosos factores socio-económicos difícilmente previsibles en el tiempo, pero fácilmente monitorizables según unos patrones determinados vinculados a la normatividad, el número de habitantes, el ahorro medio, etc. Como consecuencia, para el diseño del sistema de optimización de la vivienda crecedera, se utilizaron patrones evolutivos. Dichos patrones, alejados ya del concepto espacial y morfológico usualmente utilizado en arquitectura por figuras como C. Alexander o J. Habraken, pasaron a entenderse como una secuencia de eventos en el tiempo (espaciales, sociales, económicos, legales, etc.), que describen el proceso de transformación y que son peculiares de cada vivienda. De esta manera, el tiempo adquirió una especial importancia al convertirse en otro material más del proyecto arquitectónico. Fue en la construcción de los patrones donde se identificaron los mencionados mecanismos de ampliación, entendidos también como sistemas de compactación de la ciudad a través de la ocupación tridimensional del espacio. Al estudiar la densidad, mediante los conceptos de holgura y hacinamiento, se aceptó la congestión de las ciudades como un valor positivo. De esta forma, las posibles transformaciones realizadas por los habitantes (previstas desde un inicio) sobre el escenario del habitar (vivienda semilla), se convirtieron también en herramientas de proyecto urbano que responden a condicionantes del lugar y de los habitantes con distintas intensidades de crecimiento, ocupación y densidad. Igualmente, en el proceso de diseño del sistema de optimización, se detectaron las estrategias para la adaptabilidad y transformación de la casa crecedera, es decir, aquella serie de acciones encaminadas a la alteración de la vivienda para facilitar su ampliación, y que engloban desde sistemas constructivos en espera, que facilitan las costuras entre crecimiento y vivienda semilla, hasta sistemas espaciales que permiten que la casa altere su uso transformándose en un hábitat productivo o en un artefacto de renta. Así como los mecanismos de ampliación están asociados a la morfología, se descubrió que su uso es independiente de la localización, y que las estrategias de adaptabilidad de la vivienda se encuentran ligadas a sistemas constructivos o procesos de gestión vinculados a una región concreta. De esta manera, la combinación de los mecanismos con las estrategias caracterizan el proceso de evolución de la vivienda, vinculándola a unos determinados condicionantes sociales, geográficos y por tanto, constructivos. Finalmente, a través de la adecuada combinación de mecanismos de ampliación y estrategias de adaptabilidad en el proyecto de la vivienda con crecimiento programado es posible optimizar su desarrollo en términos económicos, constructivos, sociales y espaciales. Como resultado, esto ayudaría no sólo a mejorar la vida de los habitantes de la vivienda semilla en términos cualitativos y cuantitativos, sino también a compactar las ciudades mediante sistemas incluyentes, ya que las casas crecederas proporcionan una mayor complejidad de usos y diversidad de relaciones sociales. ABSTRACT The growing house concept -as we currently know it- was used for the first time back in 1932 in the competition Das Wachsende Haus organized by Martin Wagner and Hans Poelzig during the International Exhibition Sonne, Luft und Haus für alle, promoted by Berlin's Tourist Office. In that competition this type of housing was defined as a basic cell or a seed house unit, and depending on the needs and capabilities of the residents it could grow by adding rooms and defining itself as a complete house unit during each growing stage. Many world-top class architects such as Walter Gropius, Bruno Taut, Erich Mendelsohn or Hans Scharoun, were part of this competition exploring a new path in the flexible housing field, the scheduled grownth. From that moment and on, in Europe -and subsequently in the USA and other developed areas- many theorical and pragmatical researchs were directed towards the growing house phenomena, coming from an initial approach related to innovation, spacial and technical innovation. Furthermore -inside the traditional architecture frame in other countries, growing houses were already tested in the XVIII century- mainly due to the size were more affordable in the Real State Market. Since the 30's decade many developing countries had to deal with massive migration movements from the countryside to cities, building large housing developments were -in many cases- formed by growing housing units. In all of these developing countries the growing house approach was drawn from a different perspective than in the developed countries. An economy of means was prioritized, the utilization of low cost construction systems and -in many cases- a guided self-construction was prioritized versus the prefabricated constructions set by specialized technics that were proposed -for instance- in the European cases. To proceed with this research, information from these -and other- housing units was gathered. From then and on different ways to perform the growing actions were identified, according to its relative position from the seed house unit, these ways were named as addition or enlargement mechanisms indifferently utilized without adknowledging the geographic location for each house. The question of why one addition mechanism is preferred over another in any given case became the main target of this Thesis; the ellaboration of an analysis and diagnosis system for the growing house -according to certain parameters- would allow to point out which is the addition or addition process more efficient for a certain family in a particular location. As a starting point the grownth of the housing unit is directly linked to the evolution of the family unit that lives on it, so the house becomes a dynamic habitat. The complexity and the variability of the phenomena was taken into consideration related to a great number of socio-economic factors hardly able to be foreseen ahead on time but easy to be monitored according to certain patterns linked to regulation, population, average savings, etc As a consequence, to design the optimization system for the growing house, evolutionary patterns were utilized. Those patterns far away from the spatial and morphologic concept normally utilized in Architecture by characters like C. Alexander or J. Habraken, started to be understood like a sequence of events on time (spatial events, social events, economic events, legal events, etc) that describes the transformation process and that are particular for each housing unit. Therefore time became something important as another ingredient in the Architectural Project. The before mentioned addition or enlargement mechanisms were identified while building the patterns; these mechanisms were also understood as city's system of compactation through the tridimendional ocupation of space. Studying density, thorough the concepts of comfort and overcrowding, traffic congestion in the city was accepted as a positive value. This way, the possible transformations made by the residents (planned from the begining) about the residencial scenary (seed house), also became tools of the urban project that are a response to site's distinctive features and to the residents with different grownth intensities, activities and density Likewise, during the process of designing the optimization system, strategies for adaptations and transformation of the growing house were detected, in other words, the serial chain of actions directed to modify the house easing its enlargement or addition, and that comprehends from constructive systems on hold -that smooths the costures between grownth and housing seed- to spatial systems that allows that the house modify its utilization, becoming a productive habitat or a rental asset. Because the enlargement mechanisms are linked to the morphology, it was discovered that the use it's not related to the location, and that the adaptation strategies of the houses are linked to constructive systems or management processes linked to a particular area. This way the combination of mechanisms and strategies characterizes the process of housing evolution, linking it to certain social and geographic peculiarities and therefore constructives. At last, through the certain combination of enlargement mechanisms and adaptability strategies in the housing with scheduled grownth project is possible to optimize its development in economic, constructive, social and spatial terms. As a result, this would help not only to improve the life of the seed house residents in qualitative and quantitative terms but also to compact the cities through inclusive systems, given that the growing houses provide a larger complexity of uses and social relations.

Study of OLSR protocol with Network Simulator 2 over Linux

Until a few years ago, most of the network communications were based in the wire as the physical media, but due to the advances and the maturity of the wireless communications, this is changing. Nowadays wireless communications offers fast, secure, efficient and reliable connections. Mobile communications are in expansion, clearly driven by the use of smart phones and other mobile devices, the use of laptops, etc… Besides that point, the inversion in the installation and maintenance of the physical medium is much lower than in wired communications, not only because the air has no cost, but because the installation and maintenance of the wire require a high economic cost. Besides the economic cost we find that wire is a more vulnerable medium to external threats such as noise, sabotages, etc… There are two different types of wireless networks: those which the structure is part of the network itself and those which have a lack of structure or any centralization, in a way that the devices that form part of the network can connect themselves in a dynamic and random way, handling also the routing of every control and information messages, this kind of networks is known as Ad-hoc. In the present work we will proceed to study one of the multiple wireless protocols that allows mobile communications, it is Optimized Link State Routing, from now on, OLSR, it is an pro-active routing, standard mechanism that works in a distributed in order to stablish the connections among the different nodes that belong to a wireless network. Thanks to this protocol it is possible to get all the routing tables in all the devices correctly updated every moment through the periodical transmission of control messages and on this way allow a complete connectivity among the devices that are part of the network and also, allow access to other external networks such as virtual private networks o Internet. This protocol could be perfectly used in environments such as airports, malls, etc… The update of the routing tables in all the devices is got thanks to the periodical transmission of control messages and finally it will offer connectivity among all the devices and the corresponding external networks. For the study of OLSR protocol we will have the help of the network simulator “Network Simulator 2”, a freeware network simulator programmed in C++ based in discrete events. This simulator is used mainly in educational and research environments and allows a very extensive range of protocols, both, wired networks protocols and wireless network protocols, what is going to be really useful to proceed to the simulation of different configurations of networks and protocols. In the present work we will also study different simulations with Network Simulator 2, in different scenarios with different configurations, wired networks, and Ad-hoc networks, where we will study OLSR Protocol. RESUMEN. Hasta hace pocos años, la mayoría de las comunicaciones de red estaban basadas en el cable como medio físico pero debido al avance y madurez alcanzados en el campo de las comunicaciones inalámbricas esto está cambiando. Hoy día las comunicaciones inalámbricas nos ofrecen conexiones veloces, seguras, eficientes y fiables. Las comunicaciones móviles se encuentran en su momento de máxima expansión, claramente impulsadas por el uso de teléfonos y demás dispositivos móviles, el uso de portátiles, etc… Además la inversión a realizar en la instalación y el mantenimiento del medio físico en las comunicaciones móviles es muchísimo menor que en comunicaciones por cable, ya no sólo porque el aire no tenga coste alguno, sino porque la instalación y mantenimiento del cable precisan de un elevado coste económico por norma. Además del coste económico nos encontramos con que es un medio más vulnerable a amenazas externas tales como el ruido, escuchas no autorizadas, sabotajes, etc… Existen dos tipos de redes inalámbricas: las constituidas por una infraestructura que forma parte más o menos de la misma y las que carecen de estructura o centralización alguna, de modo que los dispositivos que forman parte de ella pueden conectarse de manera dinámica y arbitraria entre ellos, encargándose además del encaminamiento de todos los mensajes de control e información, a este tipo de redes se las conoce como redes Ad-hoc. En el presente Proyecto de Fin de Carrera se procederá al estudio de uno de los múltiples protocolos inalámbricos que permiten comunicaciones móviles, se trata del protocolo inalámbrico Optimized Link State Routing, de ahora en adelante OLSR, un mecanismo estándar de enrutamiento pro-activo, que trabaja de manera distribuida para establecer las conexiones entre los nodos que formen parte de las redes inalámbricas Ad-hoc, las cuales carecen de un nodo central y de una infraestructura pre-existente. Gracias a este protocolo es posible conseguir que todos los equipos mantengan en todo momento las tablas de ruta actualizadas correctamente mediante la transmisión periódica de mensajes de control y así permitir una completa conectividad entre todos los equipos que formen parte de la red y, a su vez, también permitir el acceso a otras redes externas tales como redes privadas virtuales o Internet. Este protocolo sería usado en entornos tales como aeropuertos La actualización de las tablas de enrutamiento de todos los equipos se conseguirá mediante la transmisión periódica de mensajes de control y así finalmente se podrá permitir conectividad entre todos los equipos y con las correspondientes redes externas. Para el estudio del protocolo OLSR contaremos con el simulador de redes Network Simulator 2, un simulador de redes freeware programado en C++ basado en eventos discretos. Este simulador es usado principalmente en ambientes educativos y de investigación y permite la simulación tanto de protocolos unicast como multicast. El campo donde más se utiliza es precisamente en el de la investigación de redes móviles Ad-hoc. El simulador Network Simulator 2 no sólo implementa el protocolo OLSR, sino que éste implementa una amplia gama de protocolos, tanto de redes cableadas como de redes inalámbricas, lo cual va a sernos de gran utilidad para proceder a la simulación de distintas configuraciones de redes y protocolos. En el presente Proyecto de Fin de Carrera se estudiarán también diversas simulaciones con el simulador NS2 en diferentes escenarios con diversas configuraciones; redes cableadas, redes inalámbricas Ad-hoc, donde se estudiará el protocolo antes mencionado: OLSR. Este Proyecto de Fin de Carrera consta de cuatro apartados distintos: Primeramente se realizará el estudio completo del protocolo OLSR, se verán los beneficios y contrapartidas que ofrece este protocolo inalámbrico. También se verán los distintos tipos de mensajes existentes en este protocolo y unos pequeños ejemplos del funcionamiento del protocolo OLSR. Seguidamente se hará una pequeña introducción al simulador de redes Network Simulator 2, veremos la historia de este simulador, y también se hará referencia a la herramienta extra NAM, la cual nos permitirá visualizar el intercambio de paquetes que se produce entre los diferentes dispositivos de nuestras simulaciones de forma intuitiva y amigable. Se hará mención a la plataforma MASIMUM, encargada de facilitar en un entorno académico software y documentación a sus alumnos con el fin de facilitarles la investigación y la simulación de redes y sensores Ad-hoc. Finalmente se verán dos ejemplos, uno en el que se realizará una simulación entre dos PCs en un entorno Ethernet y otro ejemplo en el que se realizará una simulación inalámbrica entre cinco dispositivos móviles mediante el protocolo a estudiar, OLSR.

Instruments for analysing urban conflicts in the Global South: The case of urban transformation processes in Dakar

Physical and social transformation processes that take place in urban contexts with strong spatial growth and hardly any economic development frequently have significant adverse impacts for the affected people, which tend to be made invisible. This paper presents an analytical framework to explore different ways to approach urban transformation processes (supply side), their impacts on the set of needs of the community (demand side) and their consequences on the urban environment as a whole (context). The proposed method has been used to assess three actions related to the physical and social transformation of the largest self-made settlement in the city of Dakar, Senegal, during the 2005–2012 period. Research findings show how exogenous interests are privileged over the common good when the affected citizens are not effectively involved in decision-making processes.

The lepidopteran transposon vector, piggyBac, mediates germ-line transformation in the Mediterranean fruit fly

The piggyBac (IFP2) short inverted terminal repeat transposable element from the cabbage looper Trichoplusia ni was tested for gene transfer vector function as part of a bipartite vector–helper system in the Mediterranean fruit fly Ceratitis capitata. A piggyBac vector marked with the medfly white gene was tested with a normally regulated piggyBac transposase helper at two different concentrations in a white eye host strain. Both experiments yielded transformants at an approximate frequency of 3–5%, with a total of six lines isolated having pigmented eyes with various levels of coloration. G1 transformant siblings from each line shared at least one common integration, with several sublines having an additional second integration. For the first transformant line isolated, two integrations were determined to be stable for 15 generations. For five of the lines, a piggyBac-mediated transposition was verified by sequencing the insertion site junctions isolated by inverse PCR that identified a characteristic piggyBac TTAA target site duplication. The efficient and stable transformation of the medfly with a lepidopteran vector represents transposon function over a relatively large evolutionary distance and suggests that the piggyBac system will be functional in a broad range of insects.

Dominant transformation by mutated human ras genes in vitro requires more than 100 times higher expression than is observed in cancers

The gene-mutation-cancer hypothesis holds that mutated cellular protooncogenes, such as point-mutated proto-ras, “play a dominant part in cancer,” because they are sufficient to transform transfected mouse cell lines in vitro [Alberts, B., Bray, D., Lewis, J., Raff, M., Roberts, K. & Watson, J. D. (1994) Molecular Biology of the Cell (Garland, New York)]. However, in cells transformed in vitro mutated human ras genes are expressed more than 100-fold than in the cancers from which they are isolated. In view of the discrepancy between the very low levels of ras transcription in cancers and the very high levels in cells transformed in vitro, we have investigated the minimal level of human ras expression for transformation in vitro. Using point-mutated human ras genes recombined with different promoters from either human metallothionein-IIA or human fibronectin or from retroviruses we found dominant in vitro transformation of the mouse C3H cell line only with ras genes linked to viral promoters. These ras genes were expressed more than 120-fold higher than are native ras genes of C3H cells. The copy number of transfected ras genes ranged from 2–6 in our system. In addition, nondominant transformation was observed in a small percentage (2–7%) of C3H cells transfected with ras genes that are expressed less than 20 times higher than native C3H ras genes. Because over 90% of cells expressing ras at this moderately enhanced level were untransformed, transformation must follow either a nondominant ras mechanism or a non-ras mechanism. We conclude that the mutated, but normally expressed, ras genes found in human and animal cancers are not likely to “play a dominant part in cancer.” The conclusion that mutated ras genes are not sufficient or dominant for cancer is directly supported by recent discoveries of mutated ras in normal animals, and in benign human tissue, “which has little potential to progress” [Jen, J., Powell, S. M., Papadopoulos, N., Smith, K. J., Hamilton, S. R., Vogelstein, B. & Kinzler, K. W. (1994) Cancer Res. 54, 5523–5526]. Even the view that mutated ras is necessary for cancer is hard to reconcile with (i) otherwise indistinguishable cancers with and without ras mutations, (ii) metastases of the same human cancers with and without ras mutations, (iii) retroviral ras genes that are oncogenic without point mutations, and (iv) human tumor cells having spontaneously lost ras mutation but not tumorigencity.

In vitro selection and evolution of functional proteins by using ribosome display

We report here a system with which a correctly folded complete protein and its encoding mRNA both remain attached to the ribosome and can be enriched for the ligand-binding properties of the native protein. We have selected a single-chain fragment (scFv) of an antibody 108-fold by five cycles of transcription, translation, antigen-affinity selection, and PCR. The selected scFv fragments all mutated in vitro by acquiring up to four unrelated amino acid exchanges over the five generations, but they remained fully compatible with antigen binding. Libraries of native folded proteins can now be screened and made to evolve in a cell-free system without any transformation or constraints imposed by the host cell.

Epigallocathechin-3 Gallate Selectively Inhibits the PDGF-BB–induced Intracellular Signaling Transduction Pathway in Vascular Smooth Muscle Cells and Inhibits Transformation of sis-transfected NIH 3T3 Fibroblasts and Human Glioblastoma Cells (A172)

Enhanced activity of receptor tyrosine kinases such as the PDGF β-receptor and EGF receptor has been implicated as a contributing factor in the development of malignant and nonmalignant proliferative diseases such as cancer and atherosclerosis. Several epidemiological studies suggest that green tea may prevent the development of cancer and atherosclerosis. One of the major constituents of green tea is the polyphenol epigallocathechin-3 gallate (EGCG). In an attempt to offer a possible explanation for the anti-cancer and anti-atherosclerotic activity of EGCG, we examined the effect of EGCG on the PDGF-BB–, EGF-, angiotensin II-, and FCS-induced activation of the 44 kDa and 42 kDa mitogen-activated protein (MAP) kinase isoforms (p44mapk/p42mapk) in cultured vascular smooth muscle cells (VSMCs) from rat aorta. VSMCs were treated with EGCG (1–100 μM) for 24 h and stimulated with the above mentioned agonists for different time periods. Stimulation of the p44mapk/p42mapk was detected by the enhanced Western blotting method using phospho-specific MAP kinase antibodies that recognized the Tyr204-phosphorylated (active) isoforms. Treatment of VSMCs with 10 and 50 μM EGCG resulted in an 80% and a complete inhibition of the PDGF-BB–induced activation of MAP kinase isoforms, respectively. In striking contrast, EGCG (1–100 μM) did not influence MAP kinase activation by EGF, angiotensin II, and FCS. Similarly, the maximal effect of PDGF-BB on the c-fos and egr-1 mRNA expression as well as on intracellular free Ca2+ concentration was completely inhibited in EGCG-treated VSMCs, whereas the effect of EGF was not affected. Quantification of the immunoprecipitated tyrosine-phosphorylated PDGF-Rβ, phosphatidylinositol 3′-kinase, and phospholipase C-γ1 by the enhanced Western blotting method revealed that EGCG treatment effectively inhibits tyrosine phosphorylation of these kinases in VSMCs. Furthermore, we show that spheroid formation of human glioblastoma cells (A172) and colony formation of sis-transfected NIH 3T3 cells in semisolid agar are completely inhibited by 20–50 μM EGCG. Our findings demonstrate that EGCG is a selective inhibitor of the tyrosine phosphorylation of PDGF-Rβ and its downstream signaling pathway. The present findings may partly explain the anti-cancer and anti-atherosclerotic activity of green tea.

The cellular ecology of progressive neoplastic transformation: A clonal analysis

A comparison was made of the competence for neoplastic transformation in three different sublines of NIH 3T3 cells and multiple clonal derivatives of each. Over 90% of the neoplastic foci produced by an uncloned transformed (t-SA′) subline on a confluent background of nontransformed cells were of the dense, multilayered type, but about half of the t-SA′ clones produced only light foci in assays without background. This asymmetry apparently arose from the failure of the light focus formers to register on a background of nontransformed cells. Comparison was made of the capacity for confluence-mediated transformation between uncloned parental cultures and their clonal derivatives by using two nontransformed sublines, one of which was highly sensitive and the other relatively refractory to confluence-mediated transformation. Transformation was more frequent in the clones than in the uncloned parental cultures for both sublines. This was dramatically so in the refractory subline, where the uncloned culture showed no overt sign of transformation in serially repeated assays but increasing numbers of its clones exhibited progressive transformation. The reason for the greater susceptibility of the pure clones is apparently the suppression of transformation among the diverse membership that makes up the uncloned parental culture. Progressive selection toward increasing degrees of transformation in confluent cultures plays a major role in the development of dense focus formers, but direct induction by the constraint of confluence may contribute by heritably damaging cells. In view of our finding of increased susceptibility to transformation in clonal versus uncloned populations, expansion of some clones at the expense of others during the aging process would contribute to the marked increase of cancer with age.

Transformation of malaria parasites by the spontaneous uptake and expression of DNA from human erythrocytes

The uptake and expression of extracellular DNA has been established as a mechanism for horizontal transfer of genes between bacterial species. Such transfer can support acquisition of advantageous elements, including determinants that affect the interactions between infectious organisms and their hosts. Here we show that erythrocyte-stage Plasmodium falciparum malaria parasites spontaneously take up DNA from the host cell cytoplasm into their nuclei. We have exploited this finding to produce levels of reporter expression in P.falciparum that are substantially improved over those obtained by electroporation protocols currently used to transfect malaria parasites. Parasites were transformed to a drug-resistant state when placed into cell culture with erythrocytes containing a plasmid encoding the human dihydrofolate reductase sequence. The findings reported here suggest that the malaria genome may be continually exposed to exogenous DNA from residual nuclear material in host erythrocytes.

Genomic evidence for a complete sexual cycle in Candida albicans

Candida albicans is a diploid fungus that has become a medically important opportunistic pathogen in immunocompromised individuals. We have sequenced the C. albicans genome to 10.4-fold coverage and performed a comparative genomic analysis between C. albicans and Saccharomyces cerevisiae with the objective of assessing whether Candida possesses a genetic repertoire that could support a complete sexual cycle. Analyzing over 500 genes important for sexual differentiation in S. cerevisiae, we find many homologues of genes that are implicated in the initiation of meiosis, chromosome recombination, and the formation of synaptonemal complexes. However, others are striking in their absence. C. albicans seems to have homologues of all of the elements of a functional pheromone response pathway involved in mating in S. cerevisiae but lacks many homologues of S. cerevisiae genes for meiosis. Other meiotic gene homologues in organisms ranging from filamentous fungi to Drosophila melanogaster and Caenorhabditis elegans were also found in the C. albicans genome, suggesting potential alternative mechanisms of genetic exchange.

Transcription factor EGR-1 suppresses the growth and transformation of human HT-1080 fibrosarcoma cells by induction of transforming growth factor beta 1.

The early growth response 1 (EGR-1) gene product is a transcription factor with role in differentiation and growth. We have previously shown that expression of exogenous EGR-1 in various human tumor cells unexpectedly and markedly reduces growth and tumorigenicity and, conversely, that suppression of endogenous Egr-1 expression by antisense RNA eliminates protein expression, enhances growth, and promotes phenotypic transformation. However, the mechanism of these effects remained unknown. The promoter of human transforming growth factor beta 1 (TGF-beta 1) contains two GC-rich EGR-1 binding sites. We show that expression of EGR-1 in human HT-1080 fibrosarcoma cells uses increased secretion of biologically active TGF-beta 1 in direct proportion (rPearson = 0.96) to the amount of EGR-1 expressed and addition of recombinant human TGF-beta 1 is strongly growth-suppressive for these cells. Addition of monoclonal anti-TGF-beta 1 antibodies to EGR-1-expressing HT-1080 cells completely reverses the growth inhibitory effects of EGR-1. Reporter constructs bearing the EGR-1 binding segment of the TGF-beta 1 promoter was activated 4- to 6-fold relative to a control reporter in either HT-1080 cells that stably expressed or parental cells cotransfected with an EGR-1 expression vector. Expression of delta EGR-1, a mutant that cannot interact with the corepressors, nerve growth factor-activated factor binding proteins NAB1 and NAB2, due to deletion of the repressor domain, exhibited enhanced transactivation of 2- to 3.5-fold over that of wild-type EGR-1 showing that the reporter construct reflected the appropriate in vivo regulatory context. The EGR-1-stimulated transactivation was inhibited by expression of the Wilms tumor suppressor, a known specific DNA-binding competitor. These results indicate that EGR-1 suppresses growth of human HT-1080 fibrosarcoma cells by induction of TGF-beta 1.

Homeologous recombination and mismatch repair during transformation in Streptococcus pneumoniae: saturation of the Hex mismatch repair system.

The ability of the Hex generalized mismatch repair system to prevent recombination between partially divergent (also called homeologous) sequences during transformation in Streptococcus pneumoniae was investigated. By using as donor in transformation cloned fragments 1.7-17.5% divergent in DNA sequence from the recipient, it was observed that the Hex system prevents chromosomal integration of the least and the most divergent fragments but frequently fails to do so for other fragments. In the latter case, the Hex system becomes saturated (inhibited) due to an excess of mismatches: it is unable to repair a single mismatch located elsewhere on the chromosome. Further investigation with chromosomal donor DNA, carrying only one genetically marked divergent region, revealed that a single divergent fragment can lead to saturation of the Hex system. Increase in cellular concentration of either HexA, the MutS homologue that binds mismatches, or HexB, the MutL homologue for which the essential role in repair as yet remains obscure, was shown to restore repair ability in previously saturating conditions. Investigation of heterospecific transformation by chromosomal DNA from two related streptococcal species, Streptococcus oralis and Streptococcus mitis, also revealed complete saturation of the Hex system. Therefore the Hex system is not a barrier to interspecies recombination in S. pneumoniae. These results are discussed in light of those described for the Mut system of Escherichia coli.

Complete congruence between gene diversity estimates derived from genotypic data at enzyme and random amplified polymorphic DNA loci in black spruce.

Controversy still exists over the adaptive nature of variation of enzyme loci. In conifers, random amplified polymorphic DNAs (RAPDs) represent a class of marker loci that is unlikely to fall within or be strongly linked to coding DNA. We have compared the genetic diversity in natural populations of black spruce [Picea mariana (Mill.) B.S.P.] using genotypic data at allozyme loci and RAPD loci as well as phenotypic data from inferred RAPD fingerprints. The genotypic data for both allozymes and RAPDs were obtained from at least six haploid megagametophytes for each of 75 sexually mature individuals distributed in five populations. Heterozygosities and population fixation indices were in complete agreement between allozyme loci and RAPD loci. In black spruce, it is more likely that the similar levels of variation detected at both enzyme and RAPD loci are due to such evolutionary forces as migration and the mating system, rather than to balancing selection and overdominance. Furthermore, we show that biased estimates of expected heterozygosity and among-population differentiation are obtained when using allele frequencies derived from dominant RAPD phenotypes.

Societal transformation and climate change adaptation : PAR experiences in Portugal

Tese de doutoramento, Alterações Climáticas e Políticas de Desenvolvimento Sustentável (Sociologia), Universidade de Lisboa, Instituto de Ciências Sociais, 2016

Thrombolysis in Patients Aged over 80 Years Is Equally Effective and Safe

BACKGROUND: Despite stroke's high prevalence in the elderly, intravenous thrombolysis is licensed in Europe only for patients younger than 80 years old. We aimed to compare the functional outcomes and complication rates in patients older versus younger than 80 years old treated with intravenous thrombolysis. METHODS: A retrospective observational study of patients who received intravenous thrombolysis in a stroke unit between January 1, 2009, and June 30, 2012, was conducted. Variables were compared between 2 subgroups (≤80 and >80 years). RESULTS: Overall, 512 patients underwent intravenous thrombolysis, of which 13.1% were over 80 years. The mean age was 65.4 years in the younger subgroup and 82.9 years in the older subgroup. Prior independence rates did not differ between the subgroups. Prevalence of atrial fibrillation and cardioembolic stroke was higher in the older subgroup (P = .004 and .026). Only 3% of the elderly with atrial fibrillation were taking oral anticoagulants. Symptoms-to-needle time was lower in the older subgroup (P = .048). Stroke severity was higher in patients over 80 years (P = .026). There was significant improvement in the National Institutes of Health Stroke Scale score 7 days after intravenous thrombolysis (P < .001) in both subgroups. The proportion of patients with 3 months' favorable outcome and independence, hemorrhagic transformation, and mortality rates were similar in both subgroups. CONCLUSIONS: Elderly patients' benefits and outcomes from intravenous thrombolysis treatment were identical to the younger subgroup without excess hemorrhagic transformation or mortality. These results favor the use of intravenous thrombolysis in patients over 80 years.