1950 En torno al Museo Louisiana 1970

La tesis “1950 En torno al Museo Louisiana 1970” analiza varias obras relacionadas con el espacio doméstico, que se realizaron entre 1950 y 1970 en Dinamarca, un periodo de esplendor de la Arquitectura Moderna. Tras el aislamiento y restricciones del conflicto bélico que asoló Europa, los jóvenes arquitectos daneses, estaban deseosos por experimentar nuevas ideas de procedencia internacional, favorecidos por diferentes circunstancias, encuentran el mejor campo de ensayo en el espacio doméstico. La mejor arquitectura doméstica en Dinamarca, de aquel periodo, debe entenderse como un sistema compuesto por diferentes autores, que tienen en común muchas más similitudes que diferencias, se complementan unos a otros. Para la comprensión y el entendimiento de ello se hace necesario el estudio de varias figuras y edificios, que completen este sistema cuya investigación está escasamente desarrollada. La tesis propone un viaje para conocer los nombres de algunos de sus protagonistas, que mostraron con su trabajo, que tradición y vanguardia no estarán reñidas. El objetivo es desvelar las claves de la Modernidad Danesa, reconocer, descubrir y recuperar el legado de algunos de sus protagonistas en el ámbito doméstico, cuya lección se considera de total actualidad. Una arquitectura que asume las aportaciones extranjeras con moderación y crítica, cuya íntima relación con la tradición arquitectónica y la artesanía propias, será una de sus notas especiales. Del estudio contrastado de varios proyectos y versiones, se obtienen valores comunes entre sus autores, al igual que se descubren sus afinidades o diferencias respecto a los mismos asuntos; que permitirán comprender sus actuaciones según las referencias e influencias, y definir las variables que configuran sus espacios arquitectónicos. La línea de conexión entre los edificios elegidos será su particular relación con la naturaleza y el lugar en que se integran. La fachada, lugar donde se negociará la relación entre el interior y el paisaje, será un elemento entendido de un modo diferente en cada uno de ellos, una relación que se extenderá en todas ellas, más allá de su perímetro. La investigación se ha estructurado en seis capítulos, que van precedidos de una Introducción. En el capítulo primero, se estudian y se señalan los antecedentes, las figuras y edificios más relevantes de la Tradición Danesa, para la comprensión y el esclarecimiento de algunas de las claves de su Modernidad en el campo de la Arquitectura, que se produce con una clara intención de encontrar su propia identidad y expresión. Esta Modernidad floreciente se caracteriza por la asimilación de otras culturas extranjeras desde la moderación, con un punto de vista crítico, y encuentra sus raíces ancladas a la tradición arquitectónica y la artesanía propia, que fragua en la aparición de un ideal común con enorme personalidad y que hoy se valora como una auténtica aportación de una cultura considerada entonces periférica. Se mostrará el debate y el camino seguido por las generaciones anteriores, a las obras análizadas. Las sensibilidades por lo vernáculo y lo clásico, que aparentemente son contradictorias, dominaran el debate con la misma veracidad y respetabilidad. La llamada tercera generación por Sigfried Giedion reanudará la práctica entre lo clásico y lo vernáculo, apoyados en el espíritu del trabajo artesanal y de la tradición, con el objetivo de conocer del acto arquitectónico su “la verdad” y “la esencia original”. El capítulo segundo, analiza la casa Varming, de 1953, situada en un área residencial de Gentofte, por Eva y Nils Koppel, que reinterpreta la visión de Asplund de un paisaje interior continuación del exterior, donde rompen la caja de ladrillo macizo convencional propia de los años 30. Es el ejemplo más poderoso de la unión de tradición e innovación en su obra residencial. Sus formas sobrias entre el Funcionalismo Danés y la Modernidad se singularizarán por su abstracción y volúmenes limpios que acentúan el efecto de su geometría, prismática y brutalista. El desplazamiento de los cuerpos que lo componen, unos sobre otros, generan un ritmo, que se producirá a otras escalas, ello unido a las variaciones de sus formas y a la elección de sus materiales, ladrillo y madera, le confieren a la casa un carácter orgánico. El edificio se ancla a la tierra resolviéndose en diferentes niveles tras el estudio del lugar y su topografía. El resultado es una versión construida del paisaje, en la cual el edificio da forma al lugar y ensalza la experiencia del escenario natural. La unidad de las estructuras primitivas, parece estar presente. Constituye un ejemplo de la “La idea de Promenade de Asplund”, el proyecto ofrece diferentes recorridos, permitiendo su propia vivencia de la casa, que ofrece la posibilidad vital de decidir. El capítulo tercero trata sobre el pabellón de invitados de Niels Bohr de 1957, situado un área boscosa, en Tisvilde Hegn, fue el primer edificio del arquitecto danés Vilhelm Wohlert. Arraigado a la Tradición Danesa, representa una renovación basada en la absorción de influencias extranjeras: la Arquitectura Americana y la Tradición Japonesa. La caja de madera, posada sobre un terreno horizontal, tiene el carácter sensible de un organismo vivo, siempre cambiante según las variaciones de luz del día o temperatura. Cuando se abre, crea una prolongación del espacio interior, que se extiende a la naturaleza circundante, y se expande hacia el espacio exterior, permitiendo su movilización. Se establece una arquitectura de flujos. Hay un interés por la materia, su textura y el efecto emocional que emana. Las proporciones y dimensiones del edificio están reguladas por un módulo, que se ajusta a la medida del hombre, destacando la gran unidad del edificio. La llave se su efecto estético está en su armonía y equilibrio, que transmiten serenidad y belleza. El encuentro con la naturaleza es la lección más básica del proyecto, donde un mundo de relaciones es amable al ser humano. El capítulo cuarto, analiza el proyecto del Museo Louisiana de 1958, en Humlebæk, primer proyecto de la pareja de arquitectos daneses Jørgen Bo y Vilhelm Wohlert. La experiencia en California de Wohlert donde será visitado por Bo, será trascendental para el desarrollo de Louisiana, donde la identidad Danesa se fusiona con la asimilación de otras culturas, la arquitectura de Frank Lloyd Wright, la del área de la Bahía y la Tradición Japonesa principalmente. La idea del proyecto es la de una obra de arte integral: arquitectura, arte y paisaje, coexistirían en un mismo lugar. Diferentes recursos realzarán su carácter residencial, como el uso de los materiales propios de un entorno doméstico, la realización a la escala del hombre, el modo de usar la iluminación. Cubiertas planas que muestran su artificialidad, parecen flotar sobre galerías acristaladas, acentuarán la fuerza del plano horizontal y establecerán un recorrido en zig-zag de marcado ritmo acompasado. Ritmo que tiene que ver con la encarnación del pulso de la naturaleza, que se acompaña de juegos de luz, y de otras vibraciones materiales a diferentes escalas, imagen, que encuentra una analogía semejante en la cultura japonesa. Todo se coordina con la trama estructural, que conlleva a una construcción y proporción disciplinada. Louisiana atiende al principio de crecimiento de la naturaleza, con la que su conexión es profunda. Hay un dinamismo expresado por el despliegue del edificio, que evoca a algunos proyectos de la Tradición Japonesa. Los blancos muros tienen su propia identidad como formas en sí mismas, avanzan prolongándose fuera de la línea del vidrio, se mueven libremente siguiendo el orden estructural, acompañando al espacio que fluye, en contacto directo con la naturaleza que está en un continuo estado de flujos. Se da todo un mundo de relaciones, donde existe un dialogo entre el paisaje, arte y arquitectura. El capítulo quinto, se dedica a analizar la segunda casa del arquitecto danés Halldor Gunnløgsson, de 1959. Evoca a la Arquitectura Japonesa y Americana, pero es principalmente resultado de una fuerte voluntad y disciplina artística personal. La cubierta, plana, suspendida sobre una gran plataforma pavimentada, que continúa la sección del terreno construyendo de lugar, tiene una gran presencia y arroja una profunda sombra bajo ella. En el interior un espacio único, que se puede dividir eventualmente, discurre en torno a un cuerpo central. El espacio libre fluye, extendiéndose a través de la transparencia de sus ventanales a dos espacios contrapuestos: un patio ajardinado íntimo, que inspira calma y sosiego; y la naturaleza salvaje del mar que proyecta el color del cielo, ambos en constante estado de cambio. El proyecto se elabora de un modo rigurosamente formal, existiendo al mismo tiempo un perfecto equilibrio entre la abstracción de su estructura y su programa. La estructura de madera cuyo orden se extiende más allá de los límites de su perímetro, está formada por pórticos completos como elementos libres, queda expuesta, guardando una estrecha relación con el concepto de modernidad de Mies, equivalente a la arquitectura clásica. La preocupación por el efecto estético es máxima, nada es improvisado. Pero además la combinación de materiales y el juego de las texturas hay una cualidad táctil, cierto erotismo, que flota alrededor de ella. La precisión constructiva y su refinamiento se acercan a Mies. La experiencia del espacio arquitectónico es una vivencia global. La influencia de la arquitectura japonesa, es más conceptual que formal, revelada en un respeto por la naturaleza, la búsqueda del refinamiento a través de la moderación, la eliminación de los objetos innecesarios que distraen de la experiencia del lugar y la preocupación por la luz y la sombra, donde se establece cierto paralelismo con el oscuro mundo del invierno nórdico. Hay un entendimiento de que el espacio, en lugar de ser un objeto inmaterial definido por superficies materiales se entiende como interacciones dinámicas. El capítulo sexto. Propone un viaje para conocer algunas de las viviendas unifamiliares más interesantes que se construyeron en el periodo, que forman parte del sistema investigado. Del estudio comparado y orientado en varios temas, se obtienen diversa conclusiones propias del sistema estudiado. La maestría de la sustancia y la forma será una característica distintiva en Dinamarca, se demuestra que hay un acercamiento a la cultura de Oriente, conceptual y formal, y unos intereses comunes con cierta arquitectura Americana. Su lección nos sensibiliza hacia un sentido fortalecido de proporción, escala, materialidad, textura y peso, densidad del espacio, se valora lo táctil y lo visual, hay una sensibilidad hacia la naturaleza, hacia lo humano, hacia el paisaje, la integridad de la obra. ABSTRACT The thesis “1950 around the Louisiana Museum 1970” analyses several works related to domestic space, which were carried out between 1950 and 1970 in Denmark, a golden age of modern architecture. After the isolation and limitations brought about by the war that blighted Europe, young Danish architects were keen to experiment with ideas of an international origin, encouraged by different circumstances. They find the best field of rehearsal to be the domestic space. The best architecture of that period in Denmark should be understood as a system composed of different authors, who have in common with each other many more similarities than differences, thus complimenting each other. In the interests of understanding, the study of a range of figures and buildings is necessary so that this system, the research of which is still in its infancy, can be completed. The thesis proposes a journey of discovery through the names of some of those protagonists who were showcased through their work so that tradition and avant- garde could go hand in hand. The objective is to unveil the keys to Danish Modernity; to recognise, discover and revive the legacy of some of its protagonists in the domestic field whose lessons are seen as entirely of the present. For an architect, the taking on of modern contributions with both moderation and caution, with its intimate relationship with architectural tradition and its own craft, will be one of his hallmarks. With the study set against several projects and versions, one can derive common values among their authors. In the same way their affinities and differences in respect of the same issue emerge. This will allow an understanding of their measures in line with references and influences and enable the defining of the variables of their architectural spaces. The common line between the buildings selected will be their particular relationship with nature and the space with which they integrate. The façade, the place where the relationship between the interior and the landscape would be negotiated, wouldl be the discriminating element in a distinct way for each one of them. It is through each of these facades that this relationship would extend, and far beyond their physical perimeter. The investigation has been structured into six chapters, preceded by an introduction. The first chapter outlines and analyses the backgrounds, figures and buildings most relevant to the Danish Tradition. This is to facilitate the understanding and elucidation of some of the keys to its modernity in the field of architecture, which came about with the clear intention to discover its own identity and expression. This thriving modernity is characterized by its moderate assimilation with foreign cultures with a critical eye, and finds its roots anchored in architectural tradition and its own handcraft. It is forged in the emergence of a common ideal of enormous personality which today has come to be valued as an authentic contribution to the sphere from a culture that was formerly seen as on the peripheries. What will be demonstrated is the path taken by previous generations to these works and the debate that surrounds them. The sensibilities for both the vernacular and the classic, which at first glance may seem contradictory, will dominate the debate with the same veracity and respectability. The so-called third generation of Sigfried Giedion will revive the process between the classic and the vernacular, supported in spirit by the handcraft work and by tradition, with the objective of discovering the “truth” and the “original essence” of the architectural act. The second chapter analyzes the Varming house, built by Eva and Nils Koppel 1953, which is situated in a residential area of Gentofte. This reinterprets Asplund’s vision of an interior landscape extending to the exterior, where we see a break with the conventional sturdy brick shell of the 1930s. It is the most powerful example of the union of tradition and innovation in his their residential work. Their sober forms caught between Danish Functionalism and modernity are characterized by their abstraction and clean shapes which accentuate their prismatic and brutal geometry, The displacement of the parts of which they are composed, one over the other, generate a rhythm. This is produced to varying scales and is closely linked to its forms and the selection of materials – brick and wood – that confer an organic character to the house. The building is anchored to the earth, finding solution at different levels through the study of place and topography. The result is an adaption constructed out of the landscape, in which the building gives form to the place and celebrates the experience of the natural setting. The unity of primitive structures appears to be present. It constitutes an example of “Asplund’s Promenade Idea”. Different routes of exploration within are available to the visitor, allowing for one’s own personal experience of the house, allowing in turn for the vital chance to decide. The third chapter deals with Niels Bohr’s guest pavilion. Built in 1957, it is situated in a wooded area of Tisvilde Hegn and was the architect Vilhelm Wohlert’s first building. Rooted in the Danish Tradition, it represents a renewal based on the absorption of foreign influences: American architecture and the Japanese tradition. The wooden box, perched atop a horizontal terrain, possesses the sensitive character of the living organism, ever-changing in accordance with the variations in daylight and temperature. When opened up, it creates an elongation of the interior space which extends into the surrounding nature and it expands towards the exterior space, allowing for its mobilisation. It establishes an architecture of flux. There is interest in the material, its texture and the emotional effect it inspires. The building’s proportions and dimensions are regulated by a module, which is adjusted by hand, bringing out the great unity of the building. The key to its aesthetic effect is its harmony and equilibrium, which convey serenity and beauty. The meeting with nature is the most fundamental lesson of the project, where a world of relationships softens the personality of the human being. The fourth chapter analyzes the Louisiana Museum project of 1958 in 1958. It was the first project of the Danish architects Jørgen Bo and Vilhelm Wohlert. Wohlert’s experience in California where he was visited by Bo would be essential to the development of Louisiana, where the Danish identity is fused in assimilation with other cultures, the architecture of Frank Lloyd Wright, that of the Bahía area and principally the Japanese tradition. The idea of the project was for an integrated work of art: architecture, art and landscape, which would coexist in the same space. A range of different resources would realize the residential character, such as the use of materials taken from a domestic environment, the attainment of human scale and the manner in which light was used. Flat roof plans that show their artificiality and appear to float over glassed galleries. They accentuate the strength of the horizontal plan and establish a zigzag route of marked and measured rhythm. It is a rhythm that has to do with the incarnation of nature’s pulse, which is accompanied with plays of light, as well as material vibrations of different scales, imagery which uncovers a parallel analogy with Japanese culture. Everything is coordinated along a structural frame, which involves a disciplined construction and proportion. Louisiana cherishes nature’s principle of growth, to which its connection is profound. Here is a dynamism expressed through the disposition of the building, which evokes in some projects the Japanese tradition. The white walls possess their own identity as forms in their own right. They advance, extending beyond the line of glass, moving freely along the structural line, accompanying a space that flows and in direct contact with nature that is itself in a constant state of flux. It creates a world of relationships, where dialogue exists between the landscape, art and architecture. The fifth chapter is dedicated to analyzing the Danish architect Halldor Gunnløgsson’s second house, built in 1959. It evokes both Japanese and American architecture but is principally the result of a strong will and personal artistic discipline. The flat roof suspended above a large paved platform – itself continuing the constructed terrain of the place – has great presence and casts a heavy shadow beneath. In the interior, a single space, which can at length be divided and which flows around a central space. The space flows freely, extending through the transparency of its windows which give out onto two contrasting locations: an intimate garden patio, inspiring calm and tranquillity, and the wild nature of the sea which projects the colour of sky, both in a constant state of change. The project is realized in a rigorously formal manner. A perfect balance exists between the abstraction of his structure and his project. The wooden structure, whose order extends beyond the limits of its perimeter, is formed of complete porticos of free elements. It remains exposed, maintaining a close relationship with Mies’ concept of modernity, analogous to classical architecture. The preoccupation with the aesthetic effect is paramount and nothing is improvised. But in addition to this - the combination of materials and the play of textures - there is a tactile quality, a certain eroticism, which lingers all about. The constructive precision and its refinement are close to Mies. The experience of the architectural space is universal. The influence of Japanese architecture, more conceptual than formal, is revealed in a respect for nature. It can be seen in the search for refinement through moderation, the elimination of the superfluous object that distract from the experience of place and the preoccupation with light and shade, where a certain parallel with the dark world of the Nordic winter is established. There is an understanding that space, rather than being an immaterial object defined by material surfaces, extends instead as dynamic interactions. The sixth chapter. This proposes a journey to discover some of the unfamiliar residences of most interest which were constructed in the period, and which form part of the system being investigated. Through the study of comparison and one which is geared towards various themes, diverse conclusions are drawn regarding the system being researched. The expertise in substance and form will be a distinctive characteristic in Denmark, demonstrating an approach to the culture of the Orient, both conceptual and formal, and some common interests in certain American architecture. Its teachings sensitize us to a strengthened sense of proportion, scale, materiality, texture and weight and density of space. It values both the tactile and the visual. There is a sensitivity to nature, to the human, to the landscape and to the integrity of the work.

Immune regulation by the ST6Gal sialyltransferase

The ST6Gal sialyltransferase controls production of the Siaα2-6Galβ1-4GlcNAc (Sia6LacNAc) trisaccharide, which is the ligand for the lectin CD22. Binding of CD22 to Sia6LacNAc is implicated in regulating lymphocyte adhesion and activation. We have investigated mice that lack ST6Gal and report that they are viable, yet exhibit hallmarks of severe immunosuppression unlike CD22-deficient mice. Notably, Sia6LacNAc-deficient mice display reduced serum IgM levels, impaired B cell proliferation in response to IgM and CD40 crosslinking, and attenuated antibody production to T-independent and T-dependent antigens. Deficiency of ST6Gal was further found to alter phosphotyrosine accumulation during signal transduction from the B lymphocyte antigen receptor. These studies reveal that the ST6Gal sialyltransferase and corresponding production of the Sia6LacNAc oligosaccharide are essential in promoting B lymphocyte activation and immune function.

Selective loss of dopaminergic nigro-striatal neurons in brains of Atm-deficient mice

Ataxia-telangiectasia (AT) is a human disease caused by mutations in the ATM gene. The neural phenotype of AT includes progressive cerebellar neurodegeneration, which results in ataxia and eventual motor dysfunction. Surprisingly, mice in which the Atm gene has been inactivated lack distinct behavioral ataxia or pronounced cerebellar degeneration, the hallmarks of the human disease. To determine whether lack of the Atm protein can nonetheless lead to structural abnormalities in the brain, we compared brains from male Atm-deficient mice with male, age-matched controls. Atm-deficient mice exhibited severe degeneration of tyrosine hydroxylase-positive, dopaminergic nigro-striatal neurons, and their terminals in the striatum. This cell loss was accompanied by a large reduction in immunoreactivity for the dopamine transporter in the striatum. A reduction in dopaminergic neurons also was evident in the ventral tegmental area. This effect was selective in that the noradrenergic nucleus locus coeruleus was normal in these mice. Behaviorally, Atm-deficient mice expressed locomotor abnormalities manifested as stride-length asymmetry, which could be corrected by peripheral application of the dopaminergic precursor l-dopa. In addition, these mice were hypersensitive to the dopamine releasing drug d-amphetamine. These results indicate that ATM deficiency can severely affect dopaminergic neurons in the central nervous system and suggest possible strategies for treating this aspect of the disease.

Inhibition of the electrostatic interaction between β-amyloid peptide and membranes prevents β-amyloid-induced toxicity

The accumulation of β-amyloid peptides (Aβ) into senile plaques is one of the hallmarks of Alzheimer disease. Aggregated Aβ is toxic to cells in culture and this has been considered to be the cause of neurodegeneration that occurs in the Alzheimer disease brain. The discovery of compounds that prevent Aβ toxicity may lead to a better understanding of the processes involved and ultimately to possible therapeutic drugs. Low nanomolar concentrations of Aβ1-42 and the toxic fragment Aβ25-35 have been demonstrated to render cells more sensitive to subsequent insults as manifested by an increased sensitivity to formazan crystals following MTT (3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide) reduction. Formation of the toxic β-sheet conformation by Aβ peptides is increased by negatively charged membranes. Here we demonstrate that phloretin and exifone, dipolar compounds that decrease the effective negative charge of membranes, prevent association of Aβ1-40 and Aβ25-35 to negatively charged lipid vesicles and Aβ induced cell toxicity. These results suggest that Aβ toxicity is mediated through a nonspecific physicochemical interaction with cell membranes.

Apolipoprotein E is essential for amyloid deposition in the APPV717F transgenic mouse model of Alzheimer's disease

We quantified the amount of amyloid β-peptide (Aβ) immunoreactivity as well as amyloid deposits in a large cohort of transgenic mice overexpressing the V717F human amyloid precursor protein (APPV717F+/− TG mice) with no, one, or two mouse apolipoprotein E (Apoe) alleles at various ages. Remarkably, no amyloid deposits were found in any brain region of APPV717F+/− Apoe−/− TG mice as old as 22 mo of age, whereas age-matched APPV717F +/− Apoe+/− and Apoe+/+ TG mice display abundant amyloid deposition. The amount of Aβ immunoreactivity in the hippocampus was also markedly reduced in an Apoe gene dose-dependent manner (Apoe+/+ > Apoe+/− ≫ Apoe−/−), and no Aβ immunoreactivity was detected in the cerebral cortex of APPV717F+/− Apoe−/− TG mice at any of the time points examined. The absence of apolipoprotein E protein (apoE) dramatically reduced the amount of both Aβ1–40 and Aβ1–42 immunoreactive deposits as well as the resulting astrogliosis and microgliosis normally observed in APPV717F TG mice. ApoE immunoreactivity was detected in a subset of Aβ immunoreactive deposits and in virtually all thioflavine-S-fluorescent amyloid deposits. Because the absence of apoE alters neither the transcription or translation of the APPV717F transgene nor its processing to Aβ peptide(s), we postulate that apoE promotes both the deposition and fibrillization of Aβ, ultimately affecting clearance of protease-resistant Aβ/apoE aggregates. ApoE appears to play an essential role in amyloid deposition in brain, one of the neuropathological hallmarks of Alzheimer's disease.

Genetic instability of cancer cells is proportional to their degree of aneuploidy

Genetic and phenotypic instability are hallmarks of cancer cells, but their cause is not clear. The leading hypothesis suggests that a poorly defined gene mutation generates genetic instability and that some of many subsequent mutations then cause cancer. Here we investigate the hypothesis that genetic instability of cancer cells is caused by aneuploidy, an abnormal balance of chromosomes. Because symmetrical segregation of chromosomes depends on exactly two copies of mitosis genes, aneuploidy involving chromosomes with mitosis genes will destabilize the karyotype. The hypothesis predicts that the degree of genetic instability should be proportional to the degree of aneuploidy. Thus it should be difficult, if not impossible, to maintain the particular karyotype of a highly aneuploid cancer cell on clonal propagation. This prediction was confirmed with clonal cultures of chemically transformed, aneuploid Chinese hamster embryo cells. It was found that the higher the ploidy factor of a clone, the more unstable was its karyotype. The ploidy factor is the quotient of the modal chromosome number divided by the normal number of the species. Transformed Chinese hamster embryo cells with a ploidy factor of 1.7 were estimated to change their karyotype at a rate of about 3% per generation, compared with 1.8% for cells with a ploidy factor of 0.95. Because the background noise of karyotyping is relatively high, the cells with low ploidy factor may be more stable than our method suggests. The karyotype instability of human colon cancer cell lines, recently analyzed by Lengnauer et al. [Lengnauer, C., Kinzler, K. W. & Vogelstein, B. (1997) Nature (London) 386, 623–627], also corresponds exactly to their degree of aneuploidy. We conclude that aneuploidy is sufficient to explain genetic instability and the resulting karyotypic and phenotypic heterogeneity of cancer cells, independent of gene mutation. Because aneuploidy has also been proposed to cause cancer, our hypothesis offers a common, unique mechanism of altering and simultaneously destabilizing normal cellular phenotypes.

Transforming neural computations and representing time

Motifs of neural circuitry seem surprisingly conserved over different areas of neocortex or of paleocortex, while performing quite different sensory processing tasks. This apparent paradox may be resolved by the fact that seemingly different problems in sensory information processing are related by transformations (changes of variables) that convert one problem into another. The same basic algorithm that is appropriate to the recognition of a known odor quality, independent of the strength of the odor, can be used to recognize a vocalization (e.g., a spoken syllable), independent of whether it is spoken quickly or slowly. To convert one problem into the other, a new representation of time sequences is needed. The time that has elapsed since a recent event must be represented in neural activity. The electrophysiological hallmarks of cells that are involved in generating such a representation of time are discussed. The anatomical relationships between olfactory and auditory pathways suggest relevant experiments. The neurophysiological mechanism for the psychophysical logarithmic encoding of time duration would be of direct use for interconverting olfactory and auditory processing problems. Such reuse of old algorithms in new settings and representations is related to the way that evolution develops new biochemistry.

Altered brain neurotransmitter receptors in transgenic mice expressing a portion of an abnormal human Huntington disease gene

Loss of neurotransmitter receptors, especially glutamate and dopamine receptors, is one of the pathologic hallmarks of brains of patients with Huntington disease (HD). Transgenic mice that express exon 1 of an abnormal human HD gene (line R6/2) develop neurologic symptoms at 9–11 weeks of age through an unknown mechanism. Analysis of glutamate receptors (GluRs) in symptomatic 12-week-old R6/2 mice revealed decreases compared with age-matched littermate controls in the type 1 metabotropic GluR (mGluR1), mGluR2, mGluR3, but not the mGluR5 subtype of G protein-linked mGluR, as determined by [3H]glutamate receptor binding, protein immunoblotting, and in situ hybridization. Ionotropic α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid and kainate receptors were also decreased, while N-methyl-d-aspartic acid receptors were not different compared with controls. Other neurotransmitter receptors known to be affected in HD were also decreased in R6/2 mice, including dopamine and muscarinic cholinergic, but not γ-aminobutyric acid receptors. D1-like and D2-like dopamine receptor binding was drastically reduced to one-third of control in the brains of 8- and 12-week-old R6/2 mice. In situ hybridization indicated that mGluR and D1 dopamine receptor mRNA were altered as early as 4 weeks of age, long prior to the onset of clinical symptoms. Thus, altered expression of neurotransmitter receptors precedes clinical symptoms in R6/2 mice and may contribute to subsequent pathology.

Activation of iron regulatory protein-1 by oxidative stress in vitro

Iron regulatory protein-1 (IRP-1), a central cytoplasmic regulator of cellular iron metabolism, is rapidly activated by oxidative stress to bind to mRNA iron-responsive elements. We have reconstituted the response of IRP-1 to extracellular H2O2 in a system derived from murine B6 fibroblasts permeabilized with streptolysin-O. This procedure allows separation of the cytosol from the remainder of the cells (cell pellet). IRP-1 in the cytosolic fraction fails to be directly activated by addition of H2O2. IRP-1 activation requires the presence of a nonsoluble, possibly membrane-associated component in the cell pellet. The streptolysin-O-based in vitro system faithfully recapitulates characteristic hallmarks of IRP-1 activation by H2O2 in intact cells. We show that the H2O2-mediated activation of IRP-1 is temperature dependent and sensitive to treatment with calf intestinal alkaline phosphatase (CIAP). Although IRP-1 activation is unaffected by addition of excess ATP or GTP to this in vitro system, it is negatively affected by the nonhydrolyzable nucleotide analogs adenylyl-imidodiphosphate and guanylyl-imidophosphate and completely blocked by ATP-γS and GTP-γS. The in vitro reconstitution of this oxidative stress-induced pathway has opened a different avenue for the biochemical dissection of the regulation of mammalian iron metabolism by oxidative stress. Our data show that H2O2 must be sensed to stimulate a pathway to activate IRP-1.

Arrayed transposase-binding sequences on the ends of transposon Tn5090/Tn402

The transposon Tn5090/Tn402 encodes a 559 amino acid transposase, TniA, with a DDE motif. Gel mobility shifting and cleavage protection analysis with DNase I and hydroxyl radical probes revealed that TniA binds to multiple repeat sequences on either terminus of Tn5090/Tn402. Four of these TniA-binding 19mers occurred on the left-hand (t) end and two on the right-hand (i) end. Hydroxyl radical cleavage protection demonstrated the presence of 3–6 bp contact sequences on one face of the DNA helix. The binding pattern and organisation of repeats suggested parallels between Tn5090/Tn402 and Mu, which controls its transpositional activity in the assembly step of a higher order transpososome complex. The complex terminal structure and genes of transposase and nucleotide-binding proteins in tandem are hallmarks of the handful of Mu-like elements that are known to date.

Identification and dissection of an enhancer controlling epithelial gene expression in skin

Keratins 14 and 5 are the structural hallmarks of the basal keratinocytes of the epidermis and outer root sheath (ORS) of the hair follicle. Their genes are controlled in a tissue-specific manner and thus serve as useful tools to elucidate the regulatory mechanisms involved in keratinocyte-specific transcription. Previously we identified several keratinocyte-specific DNase I hypersensitive sites (HSs) in the 5′ regulatory sequences of the K14 gene and showed that a 700-bp regulatory domain encompassing HSs II and III can confer epidermal and ORS-specific gene expression in transgenic mice in vivo. Although HS II harbored much of the transactivation activity in vitro, it was not sufficient to restrict expression to keratinocytes in vivo. We now explore the HS III regulatory element. Surprisingly, this element on its own confers gene expression to the keratinocytes of the inner root sheath (IRS) of the hair follicle, whereas a 275-bp DNA fragment containing both HSs II and III shifts the expression from the IRS to the basal keratinocytes and ORS in vivo. Electrophoretic mobility-shift assays and mutational studies of HSs III reveal a role for CACCC-box binding proteins, Sp1 family members, and other factors adding to the list of previously described factors that are involved in keratinocyte-specific gene expression. These studies highlight a cooperative interaction of the two HSs domains and strengthen the importance of combinatorial play of transcription factors that govern keratinocyte-specific gene regulation.

Customizing for clients: developing a library liaison program from need to plan*

Building on the experiences of librarian representatives to curriculum committees in the colleges of dentistry, medicine, and nursing, the Health Science Center Libraries (HSCL) Strategic Plan recommended the formation of a Library Liaison Work Group to create a formal Library Liaison Program to serve the six Health Science Center (HSC) colleges and several affiliated centers and institutes. The work group's charge was to define the purpose and scope of the program, identify models of best practice, and recommend activities for liaisons. The work group gathered background information, performed an environmental scan, and developed a philosophy statement, a program of liaison activities focusing on seven |primary areas, and a forum for liaison communication. Hallmarks of the plan included intensive subject specialization (beyond collection development), extensive communication with users, and personal information services. Specialization was expected to promote competence, communication, confidence, comfort, and customization. Development of the program required close coordination with other strategic plan implementation teams, including teams for collection development, education, and marketing. This paper discusses the HSCL's planning process and the resulting Library Liaison Program. Although focusing on an academic health center, the planning process and liaison model may be applied to any library serving diverse, subject-specific user populations.

In vivo restoration of laminin 5 β3 expression and function in junctional epidermolysis bullosa

The blistering disorder, lethal junctional epidermolysis bullosa (JEB), can result from mutations in the LAMB3 gene, which encodes laminin 5 β3 (β3). Appropriate expression of LAMβ3 in JEB skin tissue could potentially ameliorate the symptoms of the underlying disease. To explore the utility of this therapeutic approach, primary keratinocytes from six unrelated JEB patients were transduced with a retroviral vector encoding β3 and used to regenerate human skin on severe combined immunodeficient (SCID) mice. Tissue regenerated from β3-transduced JEB keratinocytes produced phenotypically normal skin characterized by sustained β3 expression and the formation of hemidesmosomes. Additionally, β3 gene transfer corrected the distribution of a number of important basement membrane zone proteins including BPAG2, integrins β4/β1, and laminins α3/γ2. Skin produced from β3-negative (β3[−]) JEB cells mimicked the hallmarks of the disease state and did not exhibit any of the aforementioned traits. Therefore, by effecting therapeutic gene transfer to β3-deficient primary keratinocytes, it is possible to produce healthy, normal skin tissue in vivo. These data support the utility of gene therapy for JEB and highlight the potential for gene delivery in the treatment of human genetic skin disease.

Genome-wide gene expression profiles of the developing mouse hippocampus

We have analyzed the developmental molecular programs of the mouse hippocampus, a cortical structure critical for learning and memory, by means of large-scale DNA microarray techniques. Of 11,000 genes and expressed sequence tags examined, 1,926 showed dynamic changes during hippocampal development from embryonic day 16 to postnatal day 30. Gene-cluster analysis was used to group these genes into 16 distinct clusters with striking patterns that appear to correlate with major developmental hallmarks and cellular events. These include genes involved in neuronal proliferation, differentiation, and synapse formation. A complete list of the transcriptional changes has been compiled into a comprehensive gene profile database (http://BrainGenomics.Princeton.edu), which should prove valuable in advancing our understanding of the molecular and genetic programs underlying both the development and the functions of the mammalian brain.

Two mutant prion proteins expressed in cultured cells acquire biochemical properties reminiscent of the scrapie isoform.

Prion diseases are a group of fatal neurodegenerative disorders that are unique in being infectious, genetic, and sporadic in origin. Infectious cases are caused by prions, which are composed primarily of PrPSc, a posttranslationally modified isoform of the normal cellular prion protein PrPC. Inherited cases are linked to insertional or point mutations in the host gene encoding PrPC. To investigate the molecular mechanisms underlying inherited prion diseases, we have constructed stably transfected Chinese hamster ovary cells that express mouse PrPs homologous to two human PrPs associated with familial Creutzfeldt-Jakob disease. One mouse PrP molecule carries a Glu-->Lys substitution at codon 199, and the other carries an insertion of six additional octapeptide repeats between codons 51 and 90. We find that both of these mutant PrPs display several biochemical hallmarks of PrPSc when synthesized in cell culture. Unlike wild-type PrP, the mutant proteins are detergent insoluble and are relatively resistant to digestion by proteinase K, yielding an N-terminally truncated core fragment of 27-30 kDa. Pulse-chase labeling experiments demonstrate that these properties are acquired posttranslationally, and are accompanied by increased metabolic stability of the protein. Our results provide the first evidence that a molecule with properties reminiscent of PrPSc can be generated de novo in cultured cells.