Re-Ocupación y/o Re-Construcción de estructuras urbanas en desuso y su conexión con el espacio público y el paisaje

Los nuevos comportamientos urbanos nos permiten observar cada vez con más frecuencia en nuestras calles y plazas realidades que siempre habíamos considerado domésticas. Al contrario también pasa, todos los días vivimos en nuestras casas situaciones que implican relacionarnos con personas que no son de nuestro núcleo familiar. El diseño doméstico de nuestras ciudades y el urbanismo del diseño de interiores parecen herramientas oportunas en el mundo que nos ha tocado vivir. Esto nos lleva a pensar que los espacios públicos y privados son términos simplificados, definidos en base a conceptos de propiedad para organizar la ciudad. En cambio, sus usos y vivencias, su gestión y sus comportamientos se han complejizado, distorsionando la terminología convencional hasta hacerla obsoleta. En este contexto, considerado también el marco socioeconómico actual, surgen las “acciones de abajo a arriba” como nuevo paradigma o modelo de renovación urbana, que entienden la involucración del ciudadano como parte activa en el proceso de construcción de la urbe desde la misma gestación del proyecto, frente a las acciones habituales que consideran al usuario como mero receptor de las propuestas. Un ciudadano que parece estar cada vez más radicalizado y una administración que parece asustarse ante el desconcierto que el acercamiento al ciudadano puede acarrear, han ocasionado por un lado, espacios “gueto” de carácter casi anárquico y, por el otro lado, lugares tan institucionalizados que derivan en espacios asociados a la administración y ajenos al ciudadano. Por ello, se considera imprescindible la colaboración entre ambos poderes. De acuerdo con el discurso que precede, dentro de un marco comparativo, se estudian 5 supuestos seleccionados de las ciudades Madrid y Zaragoza. Madrid porque es referencia nacional e internacional en el desarrollo urbano a través de procesos ‘abajo arriba’. Zaragoza porque es una ciudad ‘media’ que históricamente no se ha definido por estrategias urbanas claras, ya sean de carácter social o institucional. Sin embargo, en el momento actual se pueden identificar planteamientos relacionados con la recuperación de construcciones y espacios vacantes que pueden ser determinantes a la hora de alcanzar equilibrios con los intensos procesos institucionales acaecidos en las dos últimas décadas. De los procesos urbanos registrados en cada lugar, desarrollados en construcciones y espacios vacantes, he seleccionado: Construcciones Vacantes Madrid |Tabacalera de Lavapiés Zaragoza | Antiguo I.E.S. Luis Buñuel y antiguo Convento de Mínimos Espacios Vacantes Madrid | Campo de Cebada [solar] Zaragoza | Patio ‘antiguo I.E.S. Luis Buñuel [espacio libre] y plaza Eduardo Ibarra [espacio libre] El proyecto de investigación ha partido de las hipótesis de partida que siguen: UNA… Las ‘acciones de abajo arriba’ o renovación desde abajo no tienen cabida como elementos urbanos aislados sino conectados entre sí, posibilitando la producción de sinergias y la construcción de la ciudad; pudiendo ser consideradas acciones de desarrollo y enlace urbano, pues su objetivo es convertirse en motores del espacio público. Cuestión que es aplicable al resto de los procesos o acciones urbanas [‘horizontales’ o ‘institucionales’] DOS… La capacidad de adaptación manifestada por las ‘acciones de abajo arriba’ implica un marco ideológico de referencia asociado a la importancia de la construcción con mínimos recursos [Re-ocupación y/o Re- Construcción de estructuras urbanas en desuso] en los procesos urbanos descritos, como vía para comprender los concepto sostenibilidad y calidad figurativa de lo construido. Cuestión que es aplicable a la recuperación de aquellos aspectos de la arquitectura que la convierten en necesaria para la sociedad. Y tiene como objetivo: Identificar modelos de sostenibilidad urbana como una estrategia que va de lo individual a lo colectivo y que se transmite fundamentalmente con la acción, mezclando la innovación y la tecnología en múltiples ámbitos, utilizando los recursos naturales e intelectuales de una manera eficiente y entendiendo la inteligencia humana y sobre todo la inteligencia colectiva, como principio y justificación. Se han analizado los siguientes aspectos: sociales [participación ciudadana e implicación de la administración pública], urbanos [conexiones con otros colectivos o espacios urbanos / transformaciones urbanas a través de los procesos de gestión utilizados], constructivos [materiales utilizados en la re-construcción de construcciones y espacios vacantes / sistemas constructivos utilizados] y los relacionados con la sostenibilidad [sostenibilidad económica / sostenibilidad de mantenimiento / sostenibilidad funcional / inteligencia colectiva] El estudio de los aspectos considerados se ha desarrollado con las herramientas metodológicas siguientes: Entrevistas abiertas a expertos: se han obtenido respuestas de 25 personas expertas [5 por cada espacio o construcción vacante] relacionadas con las acciones urbanas sostenibles, la cultura y las relaciones sociales, y que también conocen los lugares y su entorno desde los puntos de vista urbano y construido. Son ‘tipos ideales’ asociados a uno de los cinco poderes que se manifiestan en la ciudad: el poder educativo [la universidad], el poder creativo [la cultura], el poder administrativo [la política], el poder empresarial [la empresa privada] y los usuarios [un usuario activo y representativo de cada lugar elegido que haya intervenido en la gestación del proceso]. Han sido personas que conocían el tejido social y urbano de la ciudad de Zaragoza y Madrid, ya que la herramienta ‘entrevista abierta a expertos’, recoge datos y opiniones planteadas en las construcciones y espacios vacantes ubicados en Zaragoza y Madrid. Entrevistas cerradas a usuarios: como la población de usuarios que se somete a la investigación es infinita o muy grande, resulta imposible o inconveniente realizar la obtención de los datos sobre todos aquellos elementos que la forman. Por lo tanto, he decidido estudiar sólo una parte de la población que denomino ‘tipos ideales’, obteniendo respuestas de 150 usuarios [30 personas por cada espacio o construcción vacante]. La selección de grupos de personas entrevistadas, debe permitir que los resultados sean representativos de la población total de usuarios. Además, la elección de ‘tipos ideales’ se ha identificado con los vecinos de los núcleos urbanos [o barrios] en los que se ubican las construcciones o espacios vacantes analizados. Observación estructurada: recoger información a través de la observación me ha permitido conocer las actuaciones y comportamientos de los ciudadanos en el medio urbano. Esto ha facilitado el estudio del medio a nivel práctico, valorando el uso que la sociedad da a las construcciones y a los espacios vacantes analizados. Es importante posicionar la estrategia en relación con el tema de investigación propuesto. Una estrategia que dibuje un panorama plural, desarrollado a través de herramientas sociales y constructivas que permitan que la arquitectura hable de cosas parecidas a lo que interesa a la ciudadanía. Para ello, propuse un catálogo de herramientas arquitectónicas que han permitido evaluar todas las propuestas. Un contexto de estrategias comunes que han descrito con los mismos códigos las distintas actuaciones analizadas. Estas herramientas tocan diferentes campos de interés. Desde las partes más tectónicas y constructivas, hasta las más ligadas con el desarrollo urbanístico y social. Acciones de participación colectiva: Experiencias o laboratorios urbanos participados por los alumnos del grado de arquitectura de la UNIZAR y los agentes sociales. Las acciones son una herramienta propositiva. Investigar y analizar proponiendo ha permitido que el análisis del contexto pueda llegar a capas de mucha más profundidad. No se ha trabajado estableciendo jerarquías de profesores y alumnos, sino que se ha intentado posibilitar la conexión de distintos agentes que trabajan coordinadamente durante el tiempo que han durado las acciones. Por un lado esto ha permite que cada integrante haya aportado al grupo lo que mejor sabe hacer y de la misma manera, que cada uno pueda aprender aquello de lo que tenga más ganas… y reflexionar sobre determinados aspectos objeto del análisis. Una vez interpretados los resultados, obtenidos a través de las herramientas metodológicas referenciadas, se ha concluido lo que sigue: Respecto de la Hipótesis de partida UNO LAS ACCIONES ‘ABAJO ARRIBA’ han revelado que no se puede entender ningún proceso de gestión urbana fuera de la participación ciudadana. El ‘ESPACIO LIBRE’ de una ciudad necesita lugares de autogestión, espacios de cogestión, movimientos de ‘arriba abajo’ y también modelos que todavía no sabemos ni que existen. LAS ACCIONES INSTITUCIONALES ‘ARRIBA ABAJO’ han demostrado que no han presentado permeabilidad ni relación con las circulaciones de entorno. Tampoco han tenido en cuenta a otras muchas personas, ‘usuarios productores’, a las que les interesan los procesos de búsqueda y las fórmulas de conexión más interactivas. Respecto de la hipótesis de partida DOS LAS ACCIONES ‘ABAJO ARRIBA’ han revelado que el ‘derecho a la ciudad’, paradigma defendido por Lefebvre desde el cual se piensa el urbanismo ciudadano, en estos supuestos podría entenderse como el ‘derecho a la infraestructura’. El ESPACIO LIBRE es infraestructura y se quiere para infraestructurar los derechos de cada uno. Y aunque también es verdad que estas acciones son simples destellos, han hecho visible otro paradigma de gestión y propuesta urbana que puede ser predominante en un futuro próximo. LAS ACCIONES INSTITUCIONALES ‘ARRIBA ABAJO’ han revelado que las intervenciones estuvieron enfocadas únicamente a la resolución de los procesos constructivos y a la incorporación del programa como un dato ‘problema’ que era necesario resolver para evitar la afección al diseño. ABSTRACT The new urban ways of behaviour let us watch more and more frequently in our streets and squares, realities that we had always considered as domestic. This also happens the other way round. Every day we have to go through situations at home which imply relationships with people who don’t belong to our family circle. The domestic design of our cities and the urban planning of indoor design seem to be adequate tools in the world we have to live in. This leads us to think that public and private spaces are simplified terms, defined according to concepts of property in order to organise the city. On the other hand, its uses and the experiences of people, its management and ways of behaviour is now more complex, changing the conventional terminology that has become outdated. In this context, ‘bottom-up’ actions arise as a new paradigm or model of urban renewal. These actions consider the active role of social participation in the process of building up the city from the very beginning, in comparison with the former way of acting that considered the user as mere receptor of the proposals. A citizen who seems to become more and more radical, and an administration that seems to be afraid of the unknown, have created both almost anarchic ghetto spaces and, on the other hand, spaces which have been so institutionalised that derive into areas associated to the administration but alienated from the citizen. For this reason, cowork of both forces is considered as crucial. In accordance with the above mentioned ideas and within a comparative framework, five situations chosen from the cities of Madrid and Zaragoza are studied. Madrid because is a national and international reference in urban development that uses “bottom-up” processes. Zaragoza because is a “medium-size” city that, historically, has not been defined by clear social or institutional urban strategies. Nevertheless, at the present time we can identify approaches on the recovery of constructions and empty areas that may be determining for reaching a balance with the intense institutional processes that have taken place in the two last decades. From the urban processes registered in every place and developed in vacant areas and constructions, I have chosen: Vacant constructions Madrid | Lavapiés Tobacco Factory Zaragoza | Old Secondary School Luis Buñuel and old Convent of the Minimos Vacant areas Madrid | Campo de Cebada [non-built site]. Zaragoza | Old courtyard of the secondary school and Eduardo Ibarra square [free space] The research project has been issued from the following starting hypotheses: ONE… “Bottom-up actions” or renewal from below have no place as isolated urban elements but as connected parts that can produce synergies and the construction of the city, and that can also be considered as actions producing urban development and links. This can also be applied to the rest of urban processes or actions [‘horizontal’ or ‘institutional’]. TWO… The capacity of adaptation shown by “bottom-up actions” implies an ideological framework of reference which is related to the importance of construction with minimal resources (re-occupation and/or reconstruction of urban structures in disuse) in the above mentioned urban processes, as a way for understanding the concepts of sustainability and the representational quality of what has been constructed. This can also be applied to the recovery of those architectural aspects that make architecture necessary for society. And its objective is: Identify models of urban sustainability as a strategy going from the individual to the collective, which are mainly transferred by action and that mix innovation and technology in many fields. Models that use natural and intellectual resources in an efficient way, and understand human intelligence and, above all, collective intelligence, as principle and justification. The following aspects have been analysed: social [civic participation and involvement of the public Administration], urban [connections with other collectives or urban spaces / urban transformation by the processes of administration used], constructive [materials used for the re-construction of empty spaces / construction systems used] and those focusing on sustainability [economic sustainability /maintenance sustainability /functional sustainability / collective intelligence]. For researching into the above mentioned aspects, the following methodological tools have been developed: Open interviews with experts: answers from 25 experts have been obtained [five for every vacant space or empty construction] on sustainable urban actions, culture and social relations, who also know the places and their environment from an urban and constructive point of view. These are “ideal types” linked to one of the five powers acting in the city: the educational power [University], the creative power [culture], the administration power [politics], the corporate power [private companies] and the users [an active and representative user for every place selected during the establishment of the process]. They were people who knew the social and urban fabric of Zaragoza and Madrid, since the “open interview for experts” tool collects data and points of view set out in vacant constructions and spaces of Zaragoza and Madrid. Close interviews with users: as the number of users targeted for the research is very big or infinite, it is impossible or inconvenient to get data from all its constituent parts. Therefore, I have decided to research into the part of the population that I call “ideal types”, obtaining answers from 150 users [30 people for every empty space or construction]. The selection of the groups of people interviewed must produce results which are representative of the total population of users. Furthermore, the election of “ideal types” has been identified with the inhabitants of urban areas [or city districts] in which the vacant spaces or constructions analysed are located. A structured observation: I have known the actions and ways of behaving of the citizens in the urban environment by means of collecting information after observation. Therefore, the practical research into the target environment has been easier by valuing the use that society gives to the empty constructions and spaces analysed. It is important to position the strategy with respect to the research subject proposed. It involves a strategy able to get an overview of a plural landscape, developed by social and constructive tools, allowing architecture to talk about topics which are interesting for city dwellers. Therefore, I proposed a set of architectural tools to evaluate all the proposals. A context of common strategies describing the different actions analysed by using the same codes. These tools focus on different fields of interests, from the most tectonic and constructive parts, to the most related to urban and social development. Actions on collective participation: experiences or urban laboratories shared by the students of architecture of the University of Zaragoza and social agents. The actions are a proactive tool. Researching and analysing by means of proposing, has allowed me to analyse the context and much deeper layers. This work has not been done by establishing ranks of professors and student, but trying to get an interaction between the different agents who work in close coordination during the implementation of the actions. This has allowed every agent to contribute the group what they do the best, and also every individual has had the possibility to learn what s/he prefers…, thinking about the different aspects targeted by the analysis. Once the different methodological tools have been interpreted, this is the conclusion: With regard to the initial hypothesis ONE “BOTTOM-UP” ACTIONS have proven that no process of urban management can be understood outside civic participation. The “FREE SPACE” of a city needs self-managed places, co-managed spaces, “up-bottom” movements, and also models whose existence is still ignored. “UP-BOTTOM” INSTITUTIONAL ACTIONS have proven that they have not presented neither permeability nor relation with local ideas. They have also disregarded many other people, the “usersproducers”, who are interested in the most interactive means of searching and connecting processes. With regard to the initial hypothesis TWO Under these premises, “BOTTOM-UP” ACTIONS have shown that the “right to the city”, a paradigm defended by Lefebvre and from which citizen-focused urbanism is conceived, could be considered as a “right to the infrastructures”. A FREE SPACE is an infrastructure and must be used to “infrastructure” the rights of every citizen. And, even though it is true that these actions are mere flashes, they have made visible another paradigm of management and urban proposal that can be prevailing in a near future. “UP-BOTTOM” INSTITUTIONAL ACTIONS have revealed that the interventions have only focused on resolving construction processes and the incorporation of the program as a “problem” data that was necessary to resolve in order to avoid its influence on the design.

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.

Evidence for balancing selection operating at the het-c heterokaryon incompatibility locus in a group of filamentous fungi

In filamentous fungi, het loci (for heterokaryon incompatibility) are believed to regulate self/nonself-recognition during vegetative growth. As filamentous fungi grow, hyphal fusion occurs within an individual colony to form a network. Hyphal fusion can occur also between different individuals to form a heterokaryon, in which genetically distinct nuclei occupy a common cytoplasm. However, heterokaryotic cells are viable only if the individuals involved have identical alleles at all het loci. One het locus, het-c, has been characterized at the molecular level in Neurospora crassa and encodes a glycine-rich protein. In an effort to understand the role of this locus in filamentous fungi, we chose to study its evolution by analyzing het-c sequence variability in species within Neurospora and related genera. We determined that the het-c locus was polymorphic in a field population of N. crassa with close to equal frequency of each of the three allelic types. Different species and even genera within the Sordariaceae shared het-c polymorphisms, indicating that these polymorphisms originated in an ancestral species. Finally, an analysis of the het-c specificity region shows a high occurrence of nonsynonymous substitution. The persistence of allelic lineages, the nearly equal allelic distribution within populations, and the high frequency of nonsynonymous substitutions in the het-c specificity region suggest that balancing selection has operated to maintain allelic diversity at het-c. Het-c shares this particular evolutionary characteristic of departing from neutrality with other self/nonself-recognition systems such as major histocompatibility complex loci in mammals and the S (self-incompatibility) locus in angiosperms.

Chrysanthemum chlorotic mottle viroid: Unusual structural properties of a subgroup of self-cleaving viroids with hammerhead ribozymes

The causal agent of chrysanthemum chlorotic mottle (CChM) disease has been identified, cloned, and sequenced. It is a viroid RNA (CChMVd) of 398–399 nucleotides. In vitro transcripts with the complete CChMVd sequence were infectious and induced the typical symptoms of the CChM disease. CChMVd can form hammerhead structures in both polarity strands. Plus and minus monomeric CChMVd RNAs self-cleaved during in vitro transcription and after purification as predicted by these structures, which are stable and most probably act as single hammerhead structures as in peach latent mosaic viroid (PLMVd), but not in avocado sunblotch viroid (ASBVd). Moreover, the plus CChMVd hammerhead structure also appears to be active in vivo, because the 5′ terminus of the linear plus CChMVd RNA isolated from infected tissue is that predicted by the corresponding hammerhead ribozyme. Both hammerhead structures of CChMVd display some peculiarities: the plus self-cleaving domain has an unpaired A after the conserved A9 residue, and the minus one has an unusually long helix II. The most stable secondary structure predicted for CChMVd is a branched conformation that does not fulfill the rod-like or quasi-rod-like model proposed for the in vitro structure of most viroids with the exception of PLMVd, whose proposed secondary structure of lowest free energy also is branched. The unusual conformation of CChMVd and PLMVd is supported by their insolubility in 2 M LiCl, in contrast to ASBVd and a series of representative non-self-cleaving viroids that are soluble under the same high salt conditions. These results support the classification of self-cleaving viroids into two subgroups, one formed by ASBVd and the other one by PLMVd and CChMVd.

Crossreactive recognition of viral, self, and bacterial peptide ligands by human class I-restricted cytotoxic T lymphocyte clonotypes: Implications for molecular mimicry in autoimmune disease

The immunodominant, CD8+ cytotoxic T lymphocyte (CTL) response to the HLA-B8-restricted peptide, RAKFKQLL, located in the Epstein–Barr virus immediate-early antigen, BZLF1, is characterized by a diverse T cell receptor (TCR) repertoire. Here, we show that this diversity can be partitioned on the basis of crossreactive cytotoxicity patterns involving the recognition of a self peptide—RSKFRQIV—located in a serine/threonine kinase and a bacterial peptide—RRKYKQII—located in Staphylococcus aureus replication initiation protein. Thus CTL clones that recognized the viral, self, and bacterial peptides expressed a highly restricted αβ TCR phenotype. The CTL clones that recognized viral and self peptides were more oligoclonal, whereas clones that strictly recognized the viral peptide displayed a diverse TCR profile. Interestingly, the self and bacterial peptides equally were substantially less effective than the cognate viral peptide in sensitizing target cell lysis, and also resulted only in a weak reactivation of memory CTLs in limiting dilution assays, whereas the cognate peptide was highly immunogenic. The described crossreactions show that human antiviral, CD8+ CTL responses can be shaped by peptide ligands derived from autoantigens and environmental bacterial antigens, thereby providing a firm structural basis for molecular mimicry involving class I-restricted CTLs in the pathogenesis of autoimmune disease.

Androgen-induced proliferative quiescence in prostate cancer cells: The role of AS3 as its mediator

In the prostate gland of adult mammals, most epithelial cells are in a state of proliferative quiescence. Androgens regulate this effect by inducing cell cycle arrest in the G0/G1 phase. Potential mediators of this androgen-induced proliferative shutoff were identified by means of subtracted cDNA libraries. The expression pattern of one of these sequences, AS3, strongly correlated with the expression of the androgen-induced proliferative shutoff both temporally and dosewise. The AS3 gene is located on chromosome 13 q12.3, in close proximity to the BRCA2 gene. The loss of chromosomal regions where AS3 alleles are located correlates with various human cancers, including prostate. The biological effect of AS3 was tested in two stable cell lines, one expressing sense and another expressing antisense AS3 constructs, both under tetracycline regulation. S9 cells were obtained by retroviral infection with virions containing a tetracycline-regulated sense AS3 construct. In these cells, sense AS3 was negatively regulated by tetracycline. Tetracycline withdrawal increased the expression of AS3 mRNA and protein. The expression of tetracycline-regulated AS3 resulted in inhibition of cell proliferation. A4 cells were obtained by retroviral infection with virions containing a tetracycline-regulated antisense AS3 construct. Vector-driven expression of antisense-AS3 blocked the induction of androgen-induced endogenous AS3 mRNA and blocked the inhibitory effect of androgens on cell proliferation. Tetracycline-regulated expression of the empty vector control had no effect on cell proliferation. These experiments strongly suggest that AS3 is a mediator of the androgen-induced proliferative shutoff.

The effect of 4,4′-diisothiocyanato-stilbene-2,2′-disulfonate on CO2 permeability of the red blood cell membrane

It has long been assumed that the red cell membrane is highly permeable to gases because the molecules of gases are small, uncharged, and soluble in lipids, such as those of a bilayer. The disappearance of 12C18O16O from a red cell suspension as the 18O exchanges between labeled CO2 + HCO3− and unlabeled HOH provides a measure of the carbonic anhydrase (CA) activity (acceleration, or A) inside the cell and of the membrane self-exchange permeability to HCO3− (Pm,HCO−3). To test this technique, we added sufficient 4,4′-diisothiocyanato-stilbene-2,2′-disulfonate (DIDS) to inhibit all the HCO3−/Cl− transport protein (Band III or capnophorin) in a red cell suspension. We found that DIDS reduced Pm,HCO−3 as expected, but also appeared to reduce intracellular A, although separate experiments showed it has no effect on CA activity in homogenous solution. A decrease in Pm,CO2 would explain this finding. With a more advanced computational model, which solves for CA activity and membrane permeabilities to both CO2 and HCO3−, we found that DIDS inhibited both Pm,HCO−3 and Pm,CO2, whereas intracellular CA activity remained unchanged. The mechanism by which DIDS reduces CO2 permeability may not be through an action on the lipid bilayer itself, but rather on a membrane transport protein, implying that this is a normal route for at least part of red cell CO2 exchange.

Medial prefrontal cortex and self-referential mental activity: Relation to a default mode of brain function

Medial prefrontal cortex (MPFC) is among those brain regions having the highest baseline metabolic activity at rest and one that exhibits decreases from this baseline across a wide variety of goal-directed behaviors in functional imaging studies. This high metabolic rate and this behavior suggest the existence of an organized mode of default brain function, elements of which may be either attenuated or enhanced. Extant data suggest that these MPFC regions may contribute to the neural instantiation of aspects of the multifaceted “self.” We explore this important concept by targeting and manipulating elements of MPFC default state activity. In this functional magnetic resonance imaging (fMRI) study, subjects made two judgments, one self-referential, the other not, in response to affectively normed pictures: pleasant vs. unpleasant (an internally cued condition, ICC) and indoors vs. outdoors (an externally cued condition, ECC). The ICC was preferentially associated with activity increases along the dorsal MPFC. These increases were accompanied by decreases in both active task conditions in ventral MPFC. These results support the view that dorsal and ventral MPFC are differentially influenced by attentiondemanding tasks and explicitly self-referential tasks. The presence of self-referential mental activity appears to be associated with increases from the baseline in dorsal MPFC. Reductions in ventral MPFC occurred consistent with the fact that attention-demanding tasks attenuate emotional processing. We posit that both self-referential mental activity and emotional processing represent elements of the default state as represented by activity in MPFC. We suggest that a useful way to explore the neurobiology of the self is to explore the nature of default state activity.

Overexpression of 5-methylcytosine DNA glycosylase in human embryonic kidney cells EcR293 demethylates the promoter of a hormone-regulated reporter gene

We have shown that the DNA demethylation complex isolated from chicken embryos has a G⋅T mismatch DNA glycosylase that also possesses 5-methylcytosine DNA glycosylase (5-MCDG) activity. Herein we show that human embryonic kidney cells stably transfected with 5-MCDG cDNA linked to a cytomegalovirus promoter overexpress 5-MCDG. A 15- to 20-fold overexpression of 5-MCDG results in the specific demethylation of a stably integrated ecdysone-retinoic acid responsive enhancer-promoter linked to a β-galactosidase reporter gene. Demethylation occurs in the absence of the ligand ponasterone A (an analogue of ecdysone). The state of methylation of the transgene was investigated by Southern blot analysis and by the bisulfite genomic sequencing reaction. Demethylation occurs downstream of the hormone response elements. No genome-wide demethylation was observed. The expression of an inactive mutant of 5-MCDG or the empty vector does not elicit any demethylation of the promoter-enhancer of the reporter gene. An increase in 5-MCDG activity does not influence the activity of DNA methyltransferase(s) when tested in vitro with a hemimethylated substrate. There is no change in the transgene copy number during selection of the clones with antibiotics. Immunoprecipitation combined with Western blot analysis showed that an antibody directed against 5-MCDG precipitates a complex containing the retinoid X receptor α. The association between retinoid receptor and 5-MCDG is not ligand dependent. These results suggest that a complex of the hormone receptor with 5-MCDG may target demethylation of the transgene in this system.

Lessons from the past: Biotic recoveries from mass extinctions

Although mass extinctions probably account for the disappearance of less than 5% of all extinct species, the evolutionary opportunities they have created have had a disproportionate effect on the history of life. Theoretical considerations and simulations have suggested that the empty niches created by a mass extinction should refill rapidly after extinction ameliorates. Under logistic models, this biotic rebound should be exponential, slowing as the environmental carrying capacity is approached. Empirical studies reveal a more complex dynamic, including positive feedback and an exponential growth phase during recoveries. Far from a model of refilling ecospace, mass extinctions appear to cause a collapse of ecospace, which must be rebuilt during recovery. Other generalities include the absence of a clear correlation between the magnitude of extinction and the pace of recovery or the resulting ecological and evolutionary disruption the presence of a survival interval, with few originations, immediately after an extinction and preceding the recovery phase, and the presence of many lineages that persist through an extinction event only to disappear during the subsequent recovery. Several recoveries include numerous missing lineages, groups that are found before the extinction, then latter in the recovery, but are missing during the initial survival–recovery phase. The limited biogeographic studies of recoveries suggest considerable variability between regions.

A loop-loop "kissing" complex is the essential part of the dimer linkage of genomic HIV-1 RNA.

RNA-RNA interactions govern a number of biological processes. Several RNAs, including natural sense and antisense RNAs, interact by means of a two-step mechanism: recognition is mediated by a loop-loop complex, which is then stabilized by formation of an extended intermolecular duplex. It was proposed that the same mechanism holds for dimerization of the genomic RNA of human immunodeficiency virus type 1 (HIV-1), an event thought to control crucial steps of HIV-1 replication. However, whereas interaction between the partially self-complementary loop of the dimerization initiation site (DIS) of each monomer is well established, formation of the extended duplex remained speculative. Here we first show that in vitro dimerization of HIV-1 RNA is a specific process, not resulting from simple annealing of denatured molecules. Next we used mutants of the DIS to test the formation of the extended duplex. Four pairs of transcomplementary mutants were designed in such a way that all pairs can form the loop-loop "kissing" complex, but only two of them can potentially form the extended duplex. All pairs of mutants form heterodimers whose thermal stability, dissociation constant, and dynamics were analyzed. Taken together, our results indicate that, in contrast with the interactions between natural sense and antisense RNAs, no extended duplex is formed during dimerization of HIV-1 RNA. We also showed that 55-mer sense RNAs containing the DIS are able to interfere with the preformed HIV-1 RNA dimer.

Expression of the CD36 homolog (FAT) in fibroblast cells: effects on fatty acid transport.

An adipocyte membrane glycoprotein, (FAT), homologous to human CD36, has been previously implicated in the binding/transport of long-chain fatty acids. It bound reactive derivatives of long-chain fatty acids and binding was specific and associated with significant inhibition of fatty acid uptake. Tissue distribution of the protein and regulation of its expression were also consistent with its postulated role. In this report, we have examined the effects of FAT expression on rates and properties of fatty acid uptake by Ob17PY fibroblasts lacking the protein. Three clones (P21, P22, and P25) were selected based on FAT mRNA and protein levels. Cell surface labeling could be demonstrated with the anti-CD36 antibody FITC-OKM5. In line with this, the major fraction of immunoreactive FAT was associated with the plasma membrane fraction. Assays of oleate and/or palmitate uptake demonstrated higher rates in the three FAT-expressing clones, compared to cells transfected with the empty vector. Clone P21, which had the highest protein levels on Western blots, exhibited the largest increase in transport rates. Fatty acid uptake in FAT-expressing P21 cells reflected two components, a phloretin-sensitive high-affinity saturable component with a Km of 0.004 microM and a basal phloretin-insensitive component that was a linear function of unbound fatty acid. P21 cells incorporated more exogenous fatty acid into phospholipids, indicating that binding of fatty acids was followed by their transfer into the cell and that both processes were increased by FAT expression. The data support the interpretation that FAT/CD36 functions as a high-affinity membrane receptor/transporter for long-chain fatty acids.

Preferential self-association of basic fibroblast growth factor is stabilized by heparin during receptor dimerization and activation.

Central to signaling by fibroblast growth factors (FGFs) is the oligomeric interaction of the growth factor and its high-affinity cell surface receptor, which is mediated by heparin-like polysaccharides. It has been proposed that the binding of heparin-like polysaccharides to FGF induces a conformational change in FGF, resulting in the formation of FGF dimers or oligomers, and this biologically active form is 'presented' to the FGF receptor for signal transduction. In this study, we show that monomeric basic FGF (FGF-2) preferentially self-associates and forms FGF-2 dimers and higher-order oligomers. As a consequence, FGF-2 monomers are oriented for binding to heparin-like polysaccharides. We also show that heparin-like polysaccharides can readily bind to self-associated FGF-2 without causing a conformational change in FGF-2 or disrupting the FGF-2 self-association, but that the bound polysaccharides only additionally stabilize the FGF-2 self-association. The preferential self-association corresponds to FGF-2 translations along two of the unit cell axes of the FGF-2 crystal structures. These two axes represent the two possible heparin binding directions, whereas the receptor binding sites are oriented along the third axis. Thus, we propose that preferential FGF-2 self-association, further stabilized by heparin, like "beads on a string," mediates FGF-2-induced receptor dimerization and activation. The observed FGF-2 self-association, modulated by heparin, not only provides a mechanism of growth factor activation but also represents a regulatory mechanism governing FGF-2 biological activity.

Introduction of the transposable element Minos into the germ line of Drosophila melanogaster.

A transposon based on the transposable element Minos from Drosophila hydei was introduced into the genome of Drosophila melanogaster using transformation mediated by the Minos transposase. The transposon carries a wild-type version of the white gene (w) of Drosophila inserted into the second exon of Minos. Transformation was obtained by injecting the transposon into preblastoderm embryos that were expressing transposase either from a Hsp70-Minos fusion inserted into the genome via P-element-mediated transformation or from a coinjected plasmid carrying the Hsp70-Minos fusion. Between 1% and 6% of the fertile injected individuals gave transformed progeny. Four of the insertions were cloned and the DNA sequences flanking the transposon ends were determined. The "empty" sites corresponding to three of the insertions were amplified from the recipient strain by PCR, cloned, and sequenced. In all cases, the transposon has inserted into a TA dinucleotide and has created the characteristic TA target site duplication. In the absence of transposase, the insertions were stable in the soma and the germ line. However, in the presence of the Hsp70-Minos gene the Minos-w transposon excises, resulting in mosaic eyes and germ-line reversion to the white phenotype. Minos could be utilized as an alternative to existing systems for transposon tagging and enhancer trapping in Drosophila; it might also be of use as a germ-line transformation vector for non-Drosophila insects.

Recoverin, a photoreceptor-specific calcium-binding protein, is expressed by the tumor of a patient with cancer-associated retinopathy.

Recoverin is a member of the EF-hand family of calcium-binding proteins involved in the transduction of light by vertebrate photoreceptors. Recoverin also was identified as an autoantigen in the degenerative disease of the retina known as cancer-associated retinopathy (CAR), a paraneoplastic syndrome whereby immunological events lead to the degeneration of photoreceptors in some individuals with cancer. In this study, we demonstrate that recoverin is expressed in the lung tumor of a CAR patient but not in similar tumors obtained from individuals without the associated retinopathy. Recoverin was identified intially by Western blot analysis of the CAR patient's biopsy tissue by using anti-recoverin antibodies generated against different regions of the recoverin molecule. In addition, cultured cells from the biopsy tissue expressed recoverin, as demonstrated by reverse transcription-PCR using RNA extracted from the cells. The immunodominant region of recoverin also was determined in this study by a solid-phase immunoassay employing overlapping heptapeptides encompassing the entire recoverin sequence. Two linear stretches of amino acids (residues 64-70, Lys-Ala-Tyr-Ala-Gln-His-Val; and 48-52, Gln-Phe-Gln-Ser-Ile) made up the major determinants. One of the same regions of the recoverin molecule (residues 64-70) also was uniquely immunopathogenic, causing photoreceptor degeneration upon immunization of Lewis rats with the corresponding peptide. These data demonstrate that the neural antigen recoverin more than likely is responsible for the immunological events associated with vision loss in some patients with cancer. These data also establish CAR as one of the few autoimmune-mediated diseases for which the specific self-antigen is known.