Functional studies of insulin-like growth factor binding protein 2 pathway in glioma progression

Comparison of gene expressing profiles between gliomas with different grades revealed frequent overexpression of insulin-like growth factor binding protein 2 (IGFBP2) in glioblastomas (GBM), in which uncontrolled cell proliferation, angiogenesis, invasion and anti-apoptosis are hallmarks. Using the glia-specific gene transfer transgenic mouse and the stable LN229(BP2) GBM cell lines, we found that IGFBP2 by itself cannot transform cells in vitro and in vivo. IGFBP2 had growth inhibitory effects on mouse primary neural progenitors, but overexpression of IGFBP2 had no effect on GBM cells. ^ Although IGFBP2 does not initiate gliomagenesis, using tissue array technology, we observed strong correlation between IGFBP2 overexpression and VEGF up-regulation in human diffuse gliomas. Furthermore, overexpression of IGFBP2 in GBM cells not only enhanced VEGF expression but also increased the malignant potential of U87 MG cells in our angiogenesis xenograft animal model. ^ In parallel to these studies, using established stable SNB19 GBM cells that overexpress IGFBP2, we found that IGFBP2 significantly increased invasion by induction of matrix metalloproteinase-2 (MMP-2) as well as other invasion related genes, providing evidence that IGFBP2 contributes to glioma progression in part by enhancing MMP-2 gene transcription and in turn tumor cell invasion. ^ Finally, we found that primary filial cells infected with an anti-sense IGFBP2 construct have markedly increased sensitivity to γ irradiation and reduced Akt activation. On the other hand, SNB19(BP2) stable lines have consistently increased levels of Akt and NFkB activation, suggesting that one possible mechanism for anti-apoptosic function of IGFBP2 is through the activation of Akt and NFkB. Beside this, what is especially interesting is the finding that Akt protein was cleaved and inactivated during apoptosis by caspases, and IGFBP2 can prevent Akt cleavage, revealing another possible mechanism through it IGFBP2 exhibit strong antiapoptotic effects. Our data showed that IGFBP2 is a specific substrate for caspase-3, raising the possibility that IGFBP2 may inhibit apoptosis by a suicide mechanism. ^ In summary, using cellular, genomics, and molecular approaches, this thesis documented the potential roles of IGFBP2 in glioma progression. Our findings shed light on an important biological aspect of glioma progression and may provide new insights useful for the design of novel mechanism-based therapies for GBM. ^

Calcium mobilization from the endoplasmic reticulum to the mitochondria during apoptosis in prostate cancer

Previous studies have implicated Ca2+ fluxes in the control of apoptosis but their exact roles in regulating the process remain obscure. Because Ca2+ can serve as a signal for cytochrome c release from isolated mitochondria, we hypothesized that alterations in intracellular Ca2+ compartmentalization might serve as a release signal in whole cells undergoing apoptosis. Exposure of human PC-3 prostate adenocarcinoma cells to staurosporine or DNA damaging agent (doxorubicin) but not to anti-Fas antibody led to early release of Ca2+ from the endoplasmic reticulum and subsequent accumulation of Ca2+ within mitochondria. Both events were blocked in cells stably transfected with Bcl-2 but were not affected by treatment with the pancaspase inhibitor, zVADfmk. The effects of staurosporine were associated with re-localization of Bax from the cytosol to both endoplasmic reticular and mitochondrial membranes. Neither ER Ca 2+ pool depletion nor mitochondrial Ca2+ uptake were observed in DU-145 cells that possess a frameshift mutation in the Bax gene unless wild-type Bax was restored via adenoviral gene transfer. Cytochrome c release and downstream features of apoptosis were attenuated by treatment with an inhibitor of mitochondria) Ca2+ uptake (RU-360). Although, direct pharmacological ER Ca2+ pool emptying in cells treated with thapsigargin did not lead to early cytochrome c release, pretreatment of cells with staurosporine dramatically sensitized mitochondria to thapsigargin-induced cytochrome c release. Together, our data demonstrate that ER-to-mitochondrial Ca2+ fluxes promote cytochrome c release and apoptosis in cells exposed to some (but not all) pro-apoptosic stimuli. ^

Whole genome analysis of Fusobacterium nucleatum, Rickettsia typhi and Francisella tularensis

The genomes of Fusobacterium nucleatum subspecies polymorphum strain ATCC 10953, Rickettsia typhi strain Wilmington, and Francisella tularensis subspecies holarctica strain OSU18 were sequenced, annotated, and analyzed. Each genome was then compared to the sequenced genomes of closely related bacteria. The genome of F. nucleatum ATCC 10953 was compared to two additional F. nucleatum subspecies, subspecies nucleatum and subspecies vincentii. This analysis revealed substantial evidence of horizontal gene transfer along with considerable genetic diversity within the species of F. nucleatum. R. typhi was compared to R. prowazekii and R. conorii. This analysis uncovered a hotspot for chromosomal rearrangements in the Spotted Fever Group but not the Typhus Group Rickettsia and revealed the close genetic relationship between the Typhus Group rickettsial species. F. tularensis OSU18 was compared to two additional F. tularensis strains. These comparisons uncovered significant chromosomal rearrangements between F. tularensis subspecies due to recombination between insertion sequence elements. ^

Tp53-dependent repression of cell cycle target genes: Global and mechanistic analysis

Most studies of p53 function have focused on genes transactivated by p53. It is less widely appreciated that p53 can repress target genes to affect a particular cellular response. There is evidence that repression is important for p53-induced apoptosis and cell cycle arrest. It is less clear if repression is important for other p53 functions. A comprehensive knowledge of the genes repressed by p53 and the cellular processes they affect is currently lacking. We used an expression profiling strategy to identify p53-responsive genes following adenoviral p53 gene transfer (Ad-p53) in PC3 prostate cancer cells. A total of 111 genes represented on the Affymetrix U133A microarray were repressed more than two fold (p ≤ 0.05) by p53. An objective assessment of array data quality was carried out using RT-PCR of 20 randomly selected genes. We estimate a confirmation rate of >95.5% for the complete data set. Functional over-representation analysis was used to identify cellular processes potentially affected by p53-mediated repression. Cell cycle regulatory genes exhibited significant enrichment (p ≤ 5E-28) within the repressed targets. Several of these genes are repressed in a p53-dependent manner following DNA damage, but preceding cell cycle arrest. These findings identify novel p53-repressed targets and indicate that p53-induced cell cycle arrest is a function of not only the transactivation of cell cycle inhibitors (e.g., p21), but also the repression of targets that act at each phase of the cell cycle. The mechanism of repression of this set of p53 targets was investigated. Most of the repressed genes identified here do not harbor consensus p53 DNA binding sites but do contain binding sites for E2F transcription factors. We demonstrate a role for E2F/RB repressor complexes in our system. Importantly, p53 is found at the promoter of CDC25A. CDC25A protein is rapidly degraded in response to DNA damage. Our group has demonstrated for the first time that CDC25A is also repressed at the transcript level by p53. This work has important implications for understanding the DNA damage cell cycle checkpoint response and the link between E2F/RB complexes and p53 in the repression of target genes. ^

Insulin-like growth factor binding protein 2 (IGFBP2) promotes glioma development and progression

Overexpression of insulin-like growth factor binding protein 2 (IGFBP2) is associated with progression and poor survival in many types of human cancer (such as prostate, ovarian, adrenocortical, breast, colorectal carcinomas, leukemia, and high-grade gliomas). We therefore hypothesize that IGFBP2 is a key regulator of tumor progression. We tested our hypothesis in gliomas using the somatic gene transfer RCAS-tva mouse model system, which permits the introduction of specific genes into specific, cell lineages, in this case glial cells (RCAS: Replication competent avian sarcomavirus, tv-a: avian RCAS virus receptor). Mice are transgenic and harbor the tv-a receptor under the control of a glial-specific promoter and study genes are cloned into the RCAS vector for post-natal intracranial delivery. For these experiments, the study genes were IGFBP2, platelet-derived growth factor B (PDGFB), K-Ras, Akt, and IIp45 (invasion inhibitory protein 45 kDa; known to bind and block IGFBP2 activity), which were delivered separately and in combination. Our results show that PDGFB signaling leads exclusively to the formation of low-grade (WHO grade II) oligodendrogliomas. PDGFB delivered in combination with IGFBP2 results in the formation of anaplastic oligodendrogliomas (WHO grade III), which are characterized by increased cellularity, vascular proliferation, small regions of necrosis, increased mitotic activity, and increased activation of the Akt pathway. IIp45 injected in combination with PDGFB and IGFBP2 ablates IGFBP2-induced tumor progression, which results in formation of low-grade oligodendrogliomas, and an overall reduction in tumor incidence. K-Ras expression was required to form astrocytomas with either IGFBP2 or Akt, indicating the activation of two separate pathways is necessary for gliomagenesis. In ex vivo experiments, blockade of Akt by an inhibitor led to decreased viability of cells co-expressing IGFBP2 versus PDGFB expression alone. This study provides definitive evidence, for the first time, that: (1) IGFBP2 plays a role in activation of the Akt pathway, (2) IGFBP2 collaborates with K-Ras or PDGFB in the development and progression of two major types of glioma, and (3) IGFBP2-induced tumor progression can be ablated by IIp45 or by specific inhibition of the Akt pathway. ^

The critical role and regulation of transcription factor FoxM1 in gastric cancer development and progression

The mammalian Forkhead Box (Fox) transcription factor (FoxM1) is implicated in tumorgenesis. However, the role and regulation of FoxM1 in gastric cancer remain unknown.^ I examined FoxM1 expression in 86 cases of primary gastric cancer and 57 normal gastric tissue specimens. I found weak expression of FoxM1 protein in normal gastric mucosa, whereas I observed strong staining for FoxM1 in tumor-cell nuclei in various gastric tumors and lymph node metastases. The aberrant FoxM1 expression is associated with VEGF expression and increased angiogenesis in human gastric cancer. A Cox proportional hazards model revealed that FoxM1 expression was an independent prognostic factor in multivariate analysis. Furthermore, overexpression of FoxM1 by gene transfer significantly promoted the growth and metastasis of gastric cancer cells in orthotopic mouse models, whereas knockdown of FoxM1 expression by small interfering RNA did the opposite. Next, I observed that alteration of tumor growth and metastasis by elevated FoxM1 expression was directly correlated with alteration of VEGF expression and angiogenesis. In addition, promotion of gastric tumorigenesis by FoxM1 directly and significantly correlated with transactivation of vascular endothelial growth factor (VEGF) expression and elevation of angiogenesis. ^ To further investigate the underlying mechanisms that result in FoxM1 overexpression in gastric cancer, I investigated FoxM1 and Krüppel-like factor 4 (KLF4) expressions in primary gastric cancer and normal gastric tissue specimens. Concomitance of increased expression of FoxM1 protein and decreased expression of KLF4 protein was evident in human gastric cancer. Enforced KLF4 expression suppressed FoxM1 protein expression. Moreover, a region within the proximal FoxM1 promoter was identified to have KLF4-binding sites. Finally, I found an increased FoxM1 expression in gastric mucosa of villin-Cre -directed tissue specific Klf4-null mice.^ In summary, I offered both clinical and mechanistic evidence that dysregulated expression of FoxM1 play an important role in gastric cancer development and progression, while KLF4 mediates negative regulation of FoxM1 expression and its loss significantly contributes to FoxM1 dysregulation. ^

THE CYTOPLASMIC TAIL OF MHC CLASS I MOLECULES PLAYS A CRITICAL ROLE IN DENDRITIC CELL-INDUCED T CELL IMMUNITY

The presentation of MHC class I (MHC-I)/peptide complexes by dendritic cells (DCs) is critical for the maintenance of central tolerance to self and for the regulation of cytotoxic T lymphocytes (CTL)-mediated adaptive immune responses against pathogens and cancer cells. Interestingly, several findings have suggested that the cytoplasmic tail of MHC class I plays a functional role in the regulation of CTL immune responses. For example, our previous studies demonstrated that exon 7-deleted MHC-I molecules not only showed extended DC cell surface half-lives but also induced significantly increased CTL responses to viral challange invivo. Although exon 7-deleted variant of MHC-I does not occur naturally in humans, the animal studies prompted us to examine whether exon 7-deleted MHC-I molecules could generate augmented CTL responses in a therapeutic DC-based vaccine setting. To examine the stimulatory capacity of exon 7-deleted MHC-I molecules, we generated a lentivirus-mediated gene transfer system to induce the expression of different MHC-I cytoplasmic tail isoforms in both mouse and human DCs. These DCs were then used as vaccines in a melanoma mouse tumor model and in a human invitro co-culture system. In this thesis, we show that DCs expressing exon 7-deleted MHC-I molecules, stimulated remarkably higher levels of T-cell cytokine production and significantly increased the proliferation of meanoma-specific (Pmel-1) T cells compared with DCs expressing wild type MHC-I. We also demonstrate that, in combination with adoptive transfer of Pmel-1 T-cell, DCs expressing exon 7-deleted Db molecules induced greater anti-tumor responses against established B16 melanoma tumors, significantly extending mouse survival as compared to DCs expressing wild-type Db molecules. Moreover, we also observed that human DCs expressing exon 7-deleted HLA-A2 molecules showed similarly augmented CTL stimulatory ability. Mechanistic studies suggest that exon 7-deleted MHC-I molecules showed impaired lateral membrane movement and extended cell surface half-lives within the DC/T-cell interface, leading to increased spatial availability of MHC-I/peptide complexes for recognition by CD8+ T cells. Collectively, these results suggesr that targeting exon 7 within the cytoplasmic tail of MHC-I molecules in DC vaccines has the potential to enhance CD8+ T cell stimulatory capacity and improve clinical outcomes in patients with cancer or viral infections.

The role of platelet -derived growth factor (PDGF) signaling in gliomagenesis

Gliomas are primary central nervous system (CNS) neoplasms that are believed to arise from astrocytes, oligodendrocytes or their precursors. Gliomas can be classified into two major histopathological groups: oligodendroglial and astroglial tumors. The most malignant of the astroglial tumors is glioblastoma multiforme (GBM). A great deal of genetic and epigenetic alterations have been implicated in gliomagenesis. In particular, PDGF signaling is frequently over-activated in a large number of human gliomas. In order to gain insights into the biology of gliomas, we manage to model human gliomas in mice using a somatic gene transfer approach—RCAS/TVA system. In our previous study, combined activation of AKT and RAS pathways gave rise to glioblastomas from CNS progenitors. In the present study, we demonstrate that in vivo autocrine PDGF stimulation induces oligodendrogliomas and mixed oligoastrocytomas from CNS progenitors and differentiated astrocytes respectively. In culture autocrine PDGF stimulation dedifferentiates astrocytes into progenitor-like cells and blockade of PDGF signaling reverses these phenotypic changes. Experimental disruption of cell cycle arrest pathway, such as Ink4a-Arf loss, is not required for the initiation of PDGF-induced gliomagenesis; instead, this mutation contributes to the tumor progression by enhancing tumor malignancy and shortening tumor latency. P53 deficiency does not promote the PDGF-induced gliomagenesis. In addition, 1p and 19q, often deleted in human oligodendrogliomas, remain intact in these PDGF-induced gliomas. Therefore, our studies suggest that autocrine PDGF stimulation alone may be sufficient to induce gliomagenesis. In contrast to transient stimulation in vitro, constitutive PDGF stimulation activates neither AKT nor RAS/MAPK pathways during gliomagenesis. This results in the formation of oligodendrogliomas, instead of glioblastomas. Sustained activation of the AKT pathway converts PDGF-induced oligodendrogliomas into astrocytomas. Our studies suggest that constitutive PDGF stimulation is not equivalent to transient PDGF stimulation, and that a transition between oligodendroglial and astroglial tumors in humans may be possible, depending on additional alterations. In summary, PDGF signaling plays a pivotal role in gliomagenesis in the mouse, and its hyperactivity is capable of contributing to both oligodendroglial and astroglial tumorigenesis. ^

Modeling, simulation and application of bacterial transduction in genetic algorithms

At present, all methods in Evolutionary Computation are bioinspired by the fundamental principles of neo-Darwinism, as well as by a vertical gene transfer. Virus transduction is one of the key mechanisms of horizontal gene propagation in microorganisms (e.g. bacteria). In the present paper, we model and simulate a transduction operator, exploring the possible role and usefulness of transduction in a genetic algorithm. The genetic algorithm including transduction has been named PETRI (abbreviation of Promoting Evolution Through Reiterated Infection). Our results showed how PETRI approaches higher fitness values as transduction probability comes close to 100%. The conclusion is that transduction improves the performance of a genetic algorithm, assuming a population divided among several sub-populations or ?bacterial colonies?.

Trans arquitectura : imaginación, invención e individuación del objeto técnico arquitectónico : transferencia tecnológica desde la industria del transporte al proyecto arquitectónico (1900-1973)

Se presenta la tesis doctoral, titulada ‘TRANS Arquitectura. Imaginación, Invención e individuación del objeto tecnico arquitectónico. Transferencia tecnológica desde la Industria del Transporte al Proyecto de Arquitectura [1900-1973]'’, que aborda la relación entre la Arquitectura y el Objeto Técnico durante la Modernidad.1 La temática de la tesis gravita en torno a la cultura técnica, la cultura material y la historia de la Tecnología del siglo XX. Hipótesis Se sostiene aquí la existencia de unas arquitecturas que se definen como Objetos Técnicos. Para demostrarlo se estudia si éstas comparten las mismas propiedades ontológicas de los objetos técnicos. Industria y Arquitectura La historia de la Arquitectura Moderna es la historia de la Industria Moderna y sus instalaciones industriales, sus productos y artefactos o sus procedimientos y procesos productivos. Fábricas, talleres, acerías, astilleros, minas, refinerías, laboratorios, automóviles, veleros, aviones, dirigibles, transbordadores, estaciones espaciales, electrodomésticos, ordenadores personales, teléfonos móviles, motores, baterías, turbinas, aparejos, cascos, chassis, carrocerías, fuselajes, composites, materiales sintéticos, la cadena de montaje, la fabricación modular, la cadena de suministros, la ingeniería de procesos, la obsolescencia programada… Todos estos objetos técnicos evolucionan constantemente gracias al inconformismo de la imaginación humana, y como intermediarios que son, cambian nuestra manera de relacionarnos con el mundo. La Arquitectura, al igual que otros objetos técnicos, media entre el hombre y el mundo. Con el objetivo de reducir el ámbito tan vasto de la investigación, éste se ha filtrado a partir de varios parámetros y cualidades de la Industria, estableciendo un marco temporal, vinculado con un determinado modo de hacer, basado en la ciencia. El inicio del desarrollo industrial basado en el conocimiento científico se da desde la Segunda Revolución Industrial, por consenso en el último tercio del siglo XIX. Este marco centra el foco de la tesis en el proceso de industrialización experimentado por la Arquitectura desde entonces, y durante aproximadamente un siglo, recorriendo la Modernidad durante los 75 primeros años del siglo XX. Durante este tiempo, los arquitectos han realizado transferencias de imágenes, técnicas, procesos y materiales desde la Industria, que ha servido como fuente de conocimiento para la Arquitectura, y ha evolucionado como disciplina. Para poder abordar más razonablemente un periodo tan amplio, se ha elegido el sector industrial del transporte, que históricamente ha sido, no sólo fuente de inspiración para los Arquitectos, sino también fuente de transferencia tecnológica para la Arquitectura. Conjuntos técnicos como los astilleros, fábricas de automóviles o hangares de aviones, individuos técnicos como barcos, coches o aviones, y elementos técnicos como las estructuras que les dan forma y soporte, son todos ellos objetos técnicos que comparten propiedades con las arquitecturas que aquí se presentan. La puesta en marcha de la cadena móvil de montaje en 1913, se toma instrumentalmente como primer foco temporal desde el que relatar la evolución de numerosos objetos técnicos en la Primera Era de la Máquina; un segundo foco se sitúa en 19582, año de la creación de la Agencia Espacial norteamericana (NASA), que sirve de referencia para situar la Segunda Era de la Máquina. La mayoría de los objetos técnicos arquitectónicos utilizados para probar la hipótesis planteada, gravitan en torno a estas fechas, con un rango de más menos 25 años, con una clara intención de sincronizar el tiempo de la acción y el tiempo del pensamiento. Arquitectura y objeto técnico Los objetos técnicos han estado siempre relacionados con la Arquitectura. En el pasado, el mismo técnico que proyectaba y supervisaba una estructura, se ocupaba de inventar los ingenios y máquinas para llevarlas a cabo. Los maestros de obra, eran verdaderos ‘agentes de transferencia tecnológica’ de la Industria y su conocimiento relacionaba técnicas de fabricación de diferentes objetos técnicos. Brunelleschi inventó varia grúas para construir la cúpula de Santa Maria dei Fiori (ca.1461), seguramente inspirado por la reedición del tratado de Vitruvio, De Architectura (15 A.C.), cuyo último capítulo estaba dedicado a las máquinas de la arquitectura clásica romana, y citaba a inventores como Archimedes. El arquitecto florentino fue el primero en patentar un invento en 1421: una embarcación anfibia que serviría para transportar mármol de Carrara por el río Arno, para su obra en Florencia. J. Paxton. Crystal Palace. London 1851. Viga-columna. Robert McCormick. Cosechadora 1831. 2ª patente, 1845. La Segunda Revolución Industrial nos dejó un primitivo ejemplo moderno de la relación entre la Arquitectura y el objeto técnico. El mayor edificio industrializado hasta la fecha, el Crystal Palace de Londres, obra de Joseph Paxton, fue montado en Londres con motivo de la Gran Exposición sobre la Industria Mundial de 1851, y siempre estará asociado a la cosechadora McCormick, merecedora del Gran Premio del Jurado. De ambos objetos técnicos, podrían destacarse características similares, como su origen industrial, y ser el complejo resultado de un ensamblaje simple de elementos técnicos. Desde la entonces, el desarrollo tecnológico ha experimentado una aceleración continuada, dando lugar a una creciente especialización y separación del conocimiento sobre las técnicas antes naturalmente unidas. Este proceso se ha dado a expensas del conocimiento integrador y en detrimento de la promiscuidad entre la Industria y la Arquitectura. Este es, sin lugar a dudas, un signo consustancial a nuestro tiempo, que provoca un natural interés de los arquitectos y otros tecnólogos, por las transferencias, trans e inter-disciplinareidades que tratan de re-establecer los canales de relación entre los diferentes campos del conocimiento. La emergencia de objetos técnicos como los vehículos modernos a principios del siglo XX (el automóvil, el trasatlántico, el dirigible o el aeroplano) está relacionada directamente con la Arquitectura de la Primera Era de la Máquina. La fascinación de los arquitectos modernos por aquellas nuevas estructuras habitables, se ha mantenido durante más de un siglo, con diferente intensidad y prestando atención a unos objetos técnicos u otros, oscilando entre el dominio del valor simbólico de los vehículos como objetosimágenes, durante el periodo heroico de la Primera Era de la Máquina, y la mirada más inquisitiva durante la Segunda, que perseguía un conocimiento más profundo de la organización de los mismos y del sistema técnico en el que estaban incluidos. La relación homóloga que existe entre arquitecturas y vehículos, por su condición de estructuras habitables, es algo de sobra conocido desde que Le Corbusier utilizara aquellas imágenes de barcos, coches y aviones para ilustrar su manifiesto Vers une architecture, de 1923. Los vehículos modernos han sido los medios con los que transmitir los conceptos que ansiaban transformar las propiedades tradicionales de la Arquitectura, relativas a su factura, su habitabilidad, su duración, su funcionalidad o su estética. Destaca particularmente el caso del automóvil en las décadas de los años 30 y 50, y los vehículos del programa espacial en las décadas de los 60 y 70. El conocimiento y la documentación previa de estos hechos, fueron un buen indicio para identificar y confirmar que el sector industrial del transporte, era un especialmente trascendente y fértil proveedor de casos de transferencia tecnológica para la Arquitectura. La tradición Moderna inaugurada por Le Corbusier en los años 20, ha sido mantenida y defendida por una multitud de arquitectos modernos como Albert Frey, Richard Neutra, Ralph Soriano, Charles Eames o Craig Ellwood, cuyo trabajo, animado por el legado de anteriores tecnólogos como Bucky Fuller o Jean Prouvé, fue fundamental y referencia obligada para la siguiente generación de arquitectos como Cedric Price, Archigram, Norman Foster, Richard Rogers, Renzo Piano, Jean Kaplicky o Richard Horden, entre otros. Todos ellos han contribuido a engrosar el imaginario del objeto técnico, aportando sus obras arquitectónicas. Estos arquitectos que aparecen repetidamente en el discurrir de la tesis, pertenecen a un mismo linaje, y son agrupados según una estructura ‘genealógica’, que se ha denominado ‘Estirpe Técnica’. Unidos por intereses comunes y similares enfoques o actitudes ante el proyecto de arquitectura, entendida como objeto Técnico, han operado mediante la práctica de la transferencia tecnológica, sin limitarse a las técnicas compositivas propias de la disciplina arquitectónica. Durante la investigación, se ha recopilado una selección de menciones explícitas -hechas por arquitectos- sobre otros objetos técnicos para referirse a la Arquitectura, mostrando las constantes y las variaciones de sus intereses a lo largo del siglo, lo que nos ha llevado a conclusiones como por ejemplo, que los conjuntos técnicos (fábricas de zepelines, aviones, automóviles o trasatlánticos) eran tomados por los arquitectos de la primera Modernidad, como un modelo imaginario, formal y compositivo, mientras que los de la Segunda Era de la Máquina los tomaban como modelo espacial y organizativo para la arquitectura. La mencionada estirpe de tecnólogos incluye líneas de descendencia conocidas, como: EiffelSuchovBehrens GropiusMiesLeCorbusierLodsProuve, en la Europa continental, o una rama británica como: LoudonPaxtonWilliamsStirlingGowan SmithsonsPriceArchigramFosterRogersPiano KaplickyHorden. También podemos encontrar conexiones intercontinentales como Fuller EamesRudolphFosterRogers, o ramificaciones menos previsibles como: LeRicolaisKahn PianoKaplicky, o LeCorbusierFreyLacaton Vassal… Seguramente muchos más merecerían incluirse en esta lista, y de hecho, la tesis asume la imposibilidad de incluirlo todo (por motivos prácticos) aunque contempla la posibilidad de ser ampliada en un futuro. Con lo aquí incluido, se pretende mostrar la continuidad en los enfoques, planteamientos y técnicas de proyectos aplicadas, de los que podemos deducir algunas conclusiones, como por ejemplo, que en los periodos inmediatamente posteriores a las dos Guerras Mundiales, aumentó la intensidad de aportaciones de nuevas imágenes de vehículos, al imaginario del objeto técnico utilizado por los arquitectos, a través de publicaciones y exposiciones. Hoy, cien años después de que Ford pusiera en marcha la cadena móvil de montaje, aún encontramos viva esta tradición en las palabras de un arquitecto, Richard Horden, cuyo trabajo porta consigo –como la información embebida en los elementos técnicos- toda una cultura técnica de una tradición moderna. Horden representa uno de los exponentes de la que he denominado estirpe de tecnólogos. Es por ello que he querido concluir la tesis con una entrevista, realizada en Mayo de 2015, en su estudio de Berkeley Square en Londres (ver Apéndices). Guías Para el desarrollo de la presente tesis, se ha tomado, como principal obra de referencia, otra tesis, titulada El modo de existencia de los objetos técnicos, leída y publicada en 1958 por el filósofo francés Gilbert Simondon [1924-89], dedicada a la ontología del objeto técnico. Esta obra enmarca el enfoque intelectual de la tesis, que entronca con la fenomenología, para movilizar una visión particular de la Arquitectura, a la que sirve como modelo de análisis ontológico para estudiar sus procesos de génesis, invención e individuación. Para el desarrollo de éstos, se ha utilizado como complemento bibliográfico, otra obra del mismo autor, titulada Imaginación e invención 1965-66. En cuanto a las fuentes historiográficas disciplinares, se ha elegido utilizar a Reyner P. Banham [1922-1988] y a Martin E. Pawley [1938-2008] como guías a través de la arquitectura del siglo XX. Sus crónicas sobre la Primera y Segunda Era de la Máquina3 y su obra crítica, han servido como índices desde los que reconstruir el imaginario del objeto técnico moderno, y del que aprovisionarse de proyectos y obras de Arquitectura como casos de estudio para la tesis. Estas obras han servido además como índices de otra bibliografía, que ha sido complementaria a la de éstos. Objetivos de la Tesis El principal objetivo de la tesis es demostrar la hipótesis: si una obra de arquitectura puede ser considerada un objeto técnico y bajo qué condiciones, construyendo un criterio que permita reconocer cuándo una obra de Arquitectura responde a la definición de objeto técnico. Otro objetivo es demostrar la importancia y potencia de la Transferencia tecnológica en el proceso evolutivo de la Arquitectura, y para ello se presentan ejemplos de una metodología de proyecto por ensamblaje, que Martin Pawley denominaba ‘Design by Assembly’. También es un objetivo el de reconstruir un Atlas del Imaginario del objeto técnico moderno, con el fin de conocer mejor las causas, razones y finalidades que llevaron a los arquitectos modernos a perseguir una arquitectura como objeto técnico. Este Atlas permite relacionar panópticamente los distintos objetos técnicos entre sí, revelando la verdadera importancia y trascendencia de aquéllos y las arquitecturas con las que se relacionan. En él, las arquitecturas vuelven a situarse en el contexto más extenso y complejo de la industria y la historia de la tecnología, al que siempre pertenecieron. De este modo, éstas son capaces de desvelar todo el conocimiento -en forma de información- que portan en su propio código ‘genético’, desplegando capítulos completos de cultura tecnológica, tan antigua como la Humanidad y en constante y creciente evolución. Estructura de la tesis Tras una Introducción en la que se presentan algunos de los conceptos principales que se instrumentalizan en la tesis sobre la ontología Simondoniana del objeto técnico y sobre la transferencia tecnológica aplicada al proyecto de Arquitectura, el texto principal de la tesis consta de tres partes: La primera se dedica a la Imaginación, una segunda parte a la Invención y una tercera a Individuación o evolución del objeto técnico. Se termina con una Discusión de la tesis y un apartado de Conclusiones. En la Introducción al objeto técnico, éste se define ontológicamente y se distinguen sus diferentes categorías (conjuntos técnicos, individuos técnicos y elementos técnicos). Se explica el proceso de génesis del objeto técnico y sus fases de imaginación, invención e individuación. También se presentan los conceptos de transducción, tecnicidad y sistema técnico, fundamentales para entender el concepto de transferencia tecnológica que se desarrollará después. La concretización, explica el modo particular de individuación y evolución de los objetos técnicos, un proceso por el que las diferentes partes de un objeto técnico, se integran y tienden hacia la propia convergencia. Aquí se comprueba la efectividad del concepto simondoniano de Transducción, como señal o información transmitida y transformada, y se relaciona con la Transferencia Tecnológica - un proceso sinergético, por el que un sector industrial se beneficia del desarrollo de otro sector- a la que se han referido explícitamente arquitectos e historiadores para explicar sus obras, durante la Segunda Era de la Máquina, y que es determinante para el desarrollo de la Industria. La transferencia tecnológica sería la transmisión del conjunto de conocimientos sobre la técnica, que incluyen su esfera fáctica, pero también la esfera sensible de la experiencia. En su aplicación a la arquitectura, las transferencias se han clasificado según tres tipos: Eidéticas, Tectónicas, Orgánicas. En la primera parte dedicada a la Imaginación del objeto técnico arquitectónico se realiza una reconstrucción ‘arqueológica’ –y parcial- del imaginario del objeto técnico moderno, con la intención de conocer mejor su génesis y la relación con otros objetos técnicos. Las fuentes de ese imaginario se buscan en las instalaciones de la Industria de principios de siglo XX, en particular en las fábricas de vehículos, con la finalidad de comprobar hasta qué punto, esos objetos técnicos fueron importantes para imaginar la Arquitectura moderna. La reconstrucción se continúa hasta la Segunda Era de la Máquina, cuando una nueva mirada más inquisitiva y precisa, se dirige a otras fábricas, vehículos y componentes, interesándose por sus cualidades materiales y organizativas. Transferencias Eidéticas, que operan desde un conocimiento intuitivo y son útiles para transmitir información sobre la esencia de un objeto técnico que sirve de fuente. Conceptos abstractos se transmiten por medio de las imágenes—objeto, para producir una transformación en su equivalente arquitectónico. Fruto de la investigación, se han detectado un grupo de conceptos que han sido objeto de transferencias tecnológicas de naturaleza eidética, provenientes del imaginario del objeto técnico moderno: FABRICADO, HABITABLE, FUNCIONAL, EFICIENTE, OBSOLESCENTE y BELLO. En la segunda parte dedicada a la Invención del objeto técnico arquitectónico, las transferencias también pueden ser Tectónicas, cuando lo que se transmite es una técnica constructiva o estructural aplicada mediante MATERIALES artificiales (como los metales, los composites como el ferrocemento, y el plywood, o las aleaciones como el aluminio) o mediante el ensamblaje de ESTRUCTURAS o partes componentes de otro objeto técnico, (como cascos, fuselajes, carrocerías o aparejos) y tiene como resultado la invención de un nuevo objeto técnico arquitectónico. En la tercera parte dedicada a la individuación, se abordan las transferencias ORGÁNICAS, lo que se transfiere es una técnica organizativa, aplicada a través de PROCEDIMIENTOS que definen la actividad del arquitecto como tecnólogo e inventor de objetos técnicos. Estos procedimientos tienen un efecto transformador en tres instituciones tradicionales para la Arquitectura: la Escuela, el Estudio y la Obra, y sus resultados se resumen en nuevos modelos de organización de la Educación de la Arquitectura, con la aparición de los Talleres de proyectos; nuevos modelos de organización del ejercicio de arquitecto: la Oficina técnica; nuevos modelos de organización del espacio, basados en la organización espacial de la Industria, que da lugar a patrones o Matrices espaciales; un nuevo modelo de organización del proyecto, que utiliza las herramientas gráficas de la industria y el ensamblaje como metodología; y un nuevo modelo de producción arquitectónica, basado en la Industrialización. Tras explicar los conceptos y la génesis del ensamblaje y el montaje, se presenta el proyecto por ensamblaje (Design by assembly) como un método que promueve la invención arquitectónica. Se demuestra utilizando algunos casos analizados en la tesis, en los que se ha realizado alguna transferencia conceptual, constructiva u organizativa. Tras analizar las arquitecturas estudiadas en la tesis, se ha utilizado el método genético propuesto por Simondon para comprender cada evolución particular, reconstruyendo las líneas genealógicas hasta sus ancestros, e identificando una serie de linajes genéticos, que corresponderían con los conjuntos técnicos estudiados en la tesis: el astillero, la fábrica de coches, y la fábrica de aeronaves: los Ancestros de la Modernidad. Los sistemas de organización espacial de estos conjuntos técnicos, están directamente relacionados con el objeto técnico que se produce en él. A partir de ellos se definen una serie de matrices operativas (MILL, SHOP, SHED), que sirven para hacer una taxonomía del objeto técnico arquitectónico. Esto se ejemplifica con algunos proyectos de Norman Foster, Richard Rogers, Renzo Piano, Nicholas Grimshaw, Jean Kaplicky y Richard Horden. Tesis: Comprobación de la hipótesis Simondon definía ontológicamente el Objeto técnico como aquello de lo que existe génesis y que desarrolla una tendencia hacia la solidaridad y unidad. Para que una Arquitectura pueda ser reconocida como un Objeto técnico, se deben dar una serie de condiciones, en las sucesivas fases que intervienen en su modo de existencia: Imaginación. Estas arquitecturas remiten a un imaginario protagonizado por imágenes-objeto de otros objetos técnicos (conjuntos técnicos, individuos técnicos y elementos técnicos). Esas imágenes-objeto vehiculizan una transferencia eidética de los objetos técnicos que simbolizan. Invención. Estas arquitecturas son el resultado de transferencias tectónicas, que se producen durante el proceso de proyecto, mediante el ensamblaje de materiales, componentes o procedimientos, utilizados en la industria para la producción de otros objetos técnicos. Individuación. Estas arquitecturas evolucionan y se individualizan por concretización, un proceso por el que los objetos técnicos se organizan para seguir su tendencia hacia la integración de sus partes, con el fin de alcanzar la convergencia de funciones en una única estructura. Esta integración tiende hacia la naturalización del objeto técnico, mediante la inclusión simbiótica de sus medios naturales asociados. En este caso, veremos cómo se ha producido transferencias orgánicas, o lo que es lo mismo, cómo los objetos técnicos –en el nivel de los conjuntos técnicos- se han tomado como modelo de organización por la arquitectura. Tras comprobar que de ellas existe una génesis, que evoluciona por las fases de imaginación e invención y concretización, se analiza su imaginario, su materialidad, sus estructuras y su organización, con el fin de detectar patrones y principios organizativos comunes a otros objetos técnicos. Interés de la tesis Desde el comienzo del nuevo siglo, diversos autores han demostrado un renovado interés por definir qué es el proyecto, qué lo constituye para qué sirve. Las aproximaciones al tema provienen de la filosofía analítica (Galle, 2008) o de la filosofía de la tecnología (Verbeek, 2005; Vermaas, 2009) y a menudo versan sobre la relación entre diseño y la cultura material (Dorschel 2003, Boradkar 2010 o Preston 2012). Es importante indicar el reciente y también creciente interés suscitado por la obra del filósofo francés, Gilbert Simondon [1924-1989], reconocida por su importante contribución a la filosofía de la técnica y la fenomenología, y por la influencia en el pensamiento de filósofos como Gilles Deleuze, autor presente en multitud de tesis doctorales e investigaciones teóricas llevadas a cabo en las principales escuelas de Arquitectura de todo el mundo desde los años 90 hasta el presente. La reedición y traducción de la obra de Simondon (ing. 1980, esp. 2008) ha recibido la atención de filósofos actuales como Paolo Virno, Bruno Latour o Bernard Stiegler, que siguen recurriendo a su estudio y análisis para avanzar en su pensamiento, estando por tanto presente en el debate contemporáneo sobre la técnica. Tras su reciente traducción al español, el pensamiento de Simondon ha despertado un gran interés en América Latina, como demuestra la organización de varios congresos y simposios, así como la proliferación de publicaciones en torno a su obra y pensamiento. Las futuras traducciones del resto de sus principales obras, asegurarán una introducción cada vez mayor en la comunidad académica. Se ha procurado presentar una mirada alternativa de la Historia de la Arquitectura Moderna, utilizando como guía a un cronista como Reyner Banham. La Era de la Máquina se ha cruzado con la Mecanología y el “vitalismo técnico” de Simondon, obteniendo como resultado una interpretación fresca, renovada y optimista de algunas de las más importantes obras de Arquitectura del siglo XX, que seguro contribuirán al desarrollo de la del siglo XXI, inmerso ya en el cambio de paradigma hacia la sostenibilidad y la ecología. ABSTRACT 'TRANS architecture. Imagination, invention and technical individuation of the architectural technical object. Technology transfer from the Transport Industry to Architectural Design [1900- 1973]' is a thesis dealing with the relationship between Architecture and the Technical Object during Modernity5. The theme of the thesis revolves around the technical culture, material culture and the history of twentieth-century technology. Hypothesis Held here is the existence of some architectures defined as technical objects. A study has been developed to prove if those architectures share the ontological properties of a technical object. Industry and Architecture The history of Modern Architecture is also the history of modern industry and its facilities, its products and devices, its procedures and production processes. Factories, workshops, steel mills, shipyards, mines, refineries, laboratories, cars, yachts, airplanes, airships, shuttles, space stations, home appliances, personal computers, mobile phones, motors, batteries, turbines, rigs, hulls, chassis, bodies, fuselages , composites and synthetic materials, the assembly line, modular manufacturing, the supply chain, process engineering, the planned obsolescence ... All these technical objects are constantly evolving thanks to the inconsistency of the human imagination and, as our intermediates, keep changing our way of relating and being in the world. Architecture, alike other technical objects, mediates between man and the World. In order to frame the vast field of the research, it has been filtered according to various parameters and qualities of Industry, establishing also a time frame which is related to a particular science-based way of making. The start of an industrial development, based on scientific knowledge is given from the Second Industrial Revolution -by consensus on the last third of the nineteenth century. This frame puts the focus of the thesis in the process of industrialization experienced by the Architecture of at least one century, and tours through Modernity during the first 75 years of the twenieth century. During this time, architects have made transfers of images, techniques, processes and materials from Industry, serving as a source of knowledge and thus allowing Architecture to evolve as a discipline. To reasonably address the enormous scope of the thesis, the industrial sector of transportation has ben chosen. It is not only a historical source of inspiration for architects, but also a traditional source of technology transfer for Modern Architecture. Technical sets such as shipyards, automobile factories or aircraft hangars, technical individuals as boats, cars or planes, and technical elements like the structures shaping and supporting them, are all technical objects which share properties with the architectures here presented. The launch of the moving assembly line in 1913, is instrumentally taken as a first time focus, from which to describe the evolution of many technical objects in the First Machine Age; a second focus could be found in 19586, year of the creation of the North American Space Agency (NASA), serving as a reference to the Second Machine Age. Most architectural technical objects used to test the hypothesis, gravitate around this second focus, in a range of plus or minus 25 years, with a clear intention to synchronize the time for action and time of thought. Architecture and Technical Object Technical objects have always been related to Architecture. In the past, the same technician who planned and oversaw a building structure, invented the devices and machines to carry them out. The foremen were the true 'technology transfer agents' from Industry. Their knowledge naturally related different manufacturing techniques to make diverse technical objects. Brunelleschi invented various cranes to build the dome of Santa Maria dei Fiori in Florence (ca.1461). Probably inspired by the reedition of Vitruvius’ treaty De Architectura (15 BC), whose last chapter was dedicated to the machines of classical Roman architecture and quoted inventors as Archimedes, the florentine architect was the first to patent an invention in 1421: an amphibious craft serving as a means of transportation for Carrara marble along the Arno river. At the daw of the Second Industrial Revolution, whose development was based on the scientific knowledge, we find a primitive modern example of the relationship between Architecture and a Technical Object: The Crystal Palace, built in London for the Great Exhibition of 1851 World Industry and designed by Joseph Paxton, was the largest to date industrialized building, and it will be always associated with the McCormick Reaper, worthy of the Grand Jury’s Prize. Similar characteristics could be emphasized of both technical objects, such as their industrial origin and for being be the complex result of a simple assembly of technical elements. Since then, technological development has experienced a continued acceleration, resulting in an increasing specialization and separation of knowledge about techniques which were naturally attached in the past. This process has happened at the expense of an integrative knowledge and against promiscuity between Industry and Architecture. This is, undoubtedly, an inherent sign of our time, which causes the natural and interest of architects and other technicians about transfers, trans-disciplinarity and inter-disciplinarity, as a reaction to reestablish channels of relationships between these different fields of knowledge. The emergence of technical objects as modern vehicles in the early twentieth century (the car, the Ocean liner, the airship or the airplane) is directly related to the Architecture of the First Machine Age. Modern architects’ fascination for those new ‘inhabitable’ structures has been maintained for over a century, with different intensity and paying attention to one and other technical objets, ranging from the domain of the symbolic value of the vehicles as objectsimages, during heroic period of the First Machine Age, to the more inquisitive glance characterizing the Second Machine Age, which sought a deeper understanding of the organization of such objects and the technical system to which they belonged. The periods immediately following both World Wars, showed a concentrated effort to bring new images of vehicles to the imaginary of architects, by means of publications and exhibitions. The homologous relationship between architectures and vehicles, in their capacity as living structures, is something well known since Le Corbusier used the images of cars, boats and airplanes to illustrate his manifesto, Towards an architecture in 1923. Modern vehicles have been the means by which to convey the concepts eager to transform the traditional attributes of Architecture: those relating to its manufacture, habitability, duration, functionality or aesthetics. The automobile stands out during the 30s and 50s, and the new vehicles of the Space Program satnd in the 60s and 70s. The prior knowledge and documentation of these events were a good indication to identify the industrial sector of Transportation as one of especial importance and as a fertile provider of technology transfer cases for Architecture. The Modern tradition, inaugurated by Le Corbusier in the 20s, has been maintained and defended by a host of modern architects like Albert Frey, Richard Neutra, Ralph Soriano, Charles Eames and Craig Ellwood, whose work - inspired by the legacy of previous technologists as Bucky Fuller or Jean Prouvé- was fundamental and a mandatory reference for the next generation of architects like Cedric Price, Archigram, Norman Foster, Richard Rogers, Renzo Piano, Jean and Richard Horden Kaplicky, among others. They have all contributed to increase the imaginary of the technical object, adding to it their architectural works. In the passage of the thesis, we repeatedly find a number of architects, who have been grouped according to a 'genealogical' structure, which has been called 'Technical Lineage'. Gathered by common interests and similar views or attitudes to the architectural design, understood as a technical object, they have operated through the practice of technology transfer, without limiting itself to specific compositional techniques of the architectural discipline. During the investigation, a selection of explicit references made by those architects, about other technical objects referring to their Architecture, has been compiled, showing constants and variations in their interests throughout the century, which has led to conclusions such as, having technicians sets (zeppelins factories, airships factories, car factories and shipyards) been taken by the architects of the first Modernity, as their main formal, compositional and imaginary models, while the Second Machine Age had taken them as a spatial and organizational model for their architecture. The above mentioned lineage of technologists includes weel-known ‘seed lines’ as: Eiffel- Suchov-Behrens, Gropius-Mies-LeCorbusier- Lods-Prouve, in continental Europe; British branches as Loudon-Paxton-Williams-Stirling- Gowan-Smithsons-Price-Archigram-Foster- Rogers-Piano-Kaplicky-Horden. And we could also find intercontinental connections as Fuller- Eames-Rudolph-Foster-Rogers, or other less predictable ramifications as LeRicolais-Kahn Piano-Kaplicky, or LeCorbusier-Frey-Lacaton & Vassal... Many more would surely deserve to be included in this list, and indeed, the thesis assumes the impossibility of including them all (for practical reasons) and even contemplates possible future extensions. The material included herein is to demonstrate the continuity in the approaches, statements and in the applied architectural design techniques, from which we can draw some conclusions. Today, one hundred years after Ford put up the moving assembly line, we still find this tradition alive in the words of the architect Richard Horden, whose work carries with it –as with the information embedded in every technical element- the whole techncial culture of a modern tradition. Horden is represented here as one of the exponents of what I have called the lineage of technologists. That is why I wanted to conclude the thesis with an interview to Richard Horden, held in May 2015 in his studio in London's Berkeley Square (see Appendices). Guides For the development of this thesis, another thesis, entitled: The mode of existence of technical objects, is taken as the main reference work. Read and published in 1958 by the French philosopher Gilbert Simondon [1924- 1989], it was dedicated to the ontology of the technical object. This work frames the intellectual approach of the thesis, which connects with phenomenology to mobilize a particular vision of Architecture. It is used as a model of ontological analysis to study its genesis, invention and evolutionary processes. To develop these, another work by the same author, titled Imagination and Invention (1965- 1966) has been used as a bibliographical complement. As for the disciplinary historical sources, Reyner P. Banham [1922-1988] and Martin E. Pawley [1938-2008] have been chosen as guides through the modern Architecture of the twentieth century. Their cronical reports on the First and Second Machine Age and their critical works have served as an index from which to reconstruct the imaginary of the modern technical object in the Machine Age7, and to stock up on projects and works of architecture, used as case studies for the thesis. These works have also been used as triggers for other literatures, which has been complementary to the former. Objectives of the Thesis The main objective of the thesis is to prove its hypothesis: if a work of architecture can be considered a technical object and under what conditions, building then a criterion for recognizing when a work of architecture meets the definition of a technical object. Another aim is to demonstrate the importance and power of Technology Transfer in the evolutionary process of Architecture, and to do it, some examples of a methodology for architectural design that Martin Pawley called 'Design by Assembly' are presented. It is also an objective to reconstruct an Atlas of the imaginary of the modern technical object, in order to better understand the causes, reasons and purposes that led modern architects to pursue architecture as a technical object. This Atlas allows to panoptically relate the various technical objects, revealing the true importance and significance of those and the architecture with whom they interact. Architectures are again at the largest and most complex industrial context and the history of technology, which always belonged. Thus, they are able to reveal all the knowledge-in the shape of information-carried in their own 'genetic' code, displaying full chapters of technological culture as old as mankind and constantly growing and evolving. Thesis: Proving the Hypothesis Simondon ontologically defined the technical object as ‘that of which genesis exists’ and that develops ‘a tendency towards solidarity and unity’. For an architecture to be recognized as a technical object, a number of conditions should be given, in the successive phases involved in their mode of existence: Imagination. These architectures refer to an imaginary featuring images-object other technical objects (technical sets, technical individuals and technical elements). These images are the means to an eidetic transfer of the technical objects which they symbolize. Invention. These architectures are the result of tectonic transfers, which occur during the architectural design process, by assembling materials, components or procedures used in industry for the production of other technical objects. Individuation. These architectures evolve and are individualized by ‘concretization’, a process leading to the full integration of its parts and aiming the full convergence of its functions into a single structure. This integration tends towards the naturalization of the technical object, by means of a symbiotic incorporation of their associated milieus. After checking if there is a genesis of them, which evolves through the phases of imagination and invention and concretization, their imaginary, materiality, structure and organization are analyzed in order to detect patterns and common organizational principles to other technical objects counterparts. Structure The main text of the thesis consists of three parts. Before there is an Introduction to the main concepts that are exploited in the thesis on ontology Simondonian technical object, and technology transfer applied to Architecture. Then a first part covers the Imaginary of the modern technical object, a second part is dedicated to the Invention and a third part to the individuation process The thesis ends with a section for the Discussion and the Conclusions. The Introduction to the technical object, this is ontologically defined and its different categories are distinguished. The process of genesis of the technical object and the phases of imagination, invention and indivuation are explained. Concepts as Transduction, Technicality and Technical system are presented for being fundamental to understand the concept of Technology Transfer that will take place later. The concretization is explained as the particular mode of individuation and evolution of technical objects, a process by which the different parts of a technical object, are integrated and begin a tendency towards a convergence in itself. The first part, dedicated to the Imagination of the architectural technical object presents a parcial "archaeological" reconstruction the imaginary of the modern technical object, intended to better understand its genesis and the relationship with other technical objects. The imaginary sources are searched in the premises of the Industry of the early twentieth century, and particularly in the factories of modern vehicles, in order to see, to what extent these technical objects were important to imagine modern architecture. The reconstruction is continued until the Second Machine Age, when a new, more inquisitive and precise gaze turns to other factories, other vehicles and other components and materials, inquiring now about their organizational qualities. The second part is devoted to the Invention of the architectural technical object. The effectiveness of the simondonian concept of Transduction is checked: a transmitted and transformed sign or information, which relates to Technology Transfer, a synergetic process by which an industrial sector benefits from the development of another sector, to which some architects and historians have explicitly referred to explain their works during Machine Age, and which is crucial for the development of the industry. Technology transfer would be the transmission of a set of information or knowledge about technique, including the factual sphere of technique, but also the sensitive sphere of experience. In their application to Architecture, these transfers have been classified according to three types: Eidetic, Tectonic and Organic. Eidetic Transfers operate from an intuitive knowledge and are useful for transmitting information about the essence of the technical object serving as a source. Abstract concepts are transmitted through the object-images to produce an equivalent transformation in Architecture. A group of concepts that have been the subject of technology transfers of eidetic nature, and have been originated in the imaginary of the modern technical object, have been detected as a result of the research: FABRICATED, INHABITABLE, FUNCTIONAL, EFFICIENT, OBSOLESCENT, and BEAUTIFUL. The transfers can also be Tectonic when, that which is transferred is a constructive or structural technique, applied through artificial MATERIALS such as metals, composites as the ferrocement, or plywood, or alloys such as aluminum; or by means of the assembly of STRUCTURES or parts of other technical objects such as hulls, fuselages, car bodies or rigs, resulting in the invention of a new architectural technical object. In the case of ORGANIC transfers, what is transferred is an organizational technique, applied by means of a set of PROCEDURES defining the activity of the architect as a technologist and inventor of technical objects. These procedures have a transformative effect on three traditional institutions for Architecture: the School, the Atelier and the Work, and the results are summarized in new models of organization of the Education of Architecture, with the onset of the Architectural Design Studios or workshops; new models of organization of the practice of architect: the technical office; and new models of space organization, based on the spatial organization of the industry, resulting in spatial patterns or spatial matrices; a new model of organization of the project, which uses graphical tools and industrail protocols as the assembly as a methodology; a new model of architectural production based on the industrialization. After explaining the concepts and the genesis of assembly and montage, Design by assembly is presented as a method that promotes architectural invention, and is shown using some case studies analyzed in the thesis, in which there has been made some conceptual, constructive or organizational transfer. After analyzing the architectures studied in the thesis, genetic method proposed by Simondon was used to understand every particular evolution, reconstructing their genealogical lines up to their ancestors, identifying a series of genetic lineages, which correspond to the technical sets studied in the thesis : the shipyard, the car factory, and aircraft factory. The real ancestors of Modernity. The spatial organization systems of these technical sets are directly related to the technical object that is fabricated within them. From that point, a number of operational matrices are defined (MILL, SHOP, SHED) and used to make a taxonomy of the architectural technical object. This is exemplified by some projects by architects as Norman Foster, Richard Rogers, Renzo Piano, Nicholas Grimshaw, Jean and Richard Horden Kaplicky. Interest of the thesis Since the beginning of the new century, several authors have shown a renewed interest in defining what a project is, how it is constituted and what it is for. The approaches to the subject are brought from analytic philosophy (Galle, 2008) or from the philosophy of technology (Verbeek, 2005; Vermaas, 2009) and they often speak about the relationship between design and material culture (Dorschel 2003, 2010 or Preston Boradkar 2012). It is also important to note the recent and growing interest in the work of French philosopher Gilbert Simondon [1924-1989], mainly known for its important contribution to the philosophy of technology and phenomenology of the technical object, and the influence on the thinking of contemporary philosophers as Paolo Virno, Bruno Latour or Gilles Deleuze, being the latter a author present in many doctoral theses and theoretical research conducted at major architecture schools around the world since the 90s to the present. The republication and translation of the work of Simondon (eng. 1980, spn. 2008) has received the attention from current philosophers such as Bernard Stiegler who continues to use its study and analysis to advance his thinking, thus being present in the contemporary debate about the technique. After its recent translation into Spanish, the thought of Simondon has aroused great interest in Latin America, as evidenced by the organization of various conferences and symposia, as well as the proliferation of publications about his work and thought8. Future translations of the rest of his major works, will ensure increased introduction in the academic community. Efforts have been made to present an alternative view of the History of Modern Architecture, using a reporter as Reyner P.Banham as a guide. The Machine Age intersects Simondon’s mechanology and his "technical vitalism", resulting in a fresh, renewed and optimistic interpretation of some of the most important works of Architecture of the twentieth century, which will surely contribute to the development of this century’s Architecture, already immersed in the paradigm shift towards sustainability and ecology.

Import of DNA into mammalian nuclei by proteins originating from a plant pathogenic bacterium

Import of DNA into mammalian nuclei is generally inefficient. Therefore, one of the current challenges in human gene therapy is the development of efficient DNA delivery systems. Here we tested whether bacterial proteins could be used to target DNA to mammalian cells. Agrobacterium tumefaciens, a plant pathogen, efficiently transfers DNA as a nucleoprotein complex to plant cells. Agrobacterium-mediated T-DNA transfer to plant cells is the only known example for interkingdom DNA transfer and is widely used for plant transformation. Agrobacterium virulence proteins VirD2 and VirE2 perform important functions in this process. We reconstituted complexes consisting of the bacterial virulence proteins VirD2, VirE2, and single-stranded DNA (ssDNA) in vitro. These complexes were tested for import into HeLa cell nuclei. Import of ssDNA required both VirD2 and VirE2 proteins. A VirD2 mutant lacking its C-terminal nuclear localization signal was deficient in import of the ssDNA–protein complexes into nuclei. Import of VirD2–ssDNA–VirE2 complexes was fast and efficient, and was shown to depended on importin α, Ran, and an energy source. We report here that the bacterium-derived and plant-adapted protein–DNA complex, made in vitro, can be efficiently imported into mammalian nuclei following the classical importin-dependent nuclear import pathway. This demonstrates the potential of our approach to enhance gene transfer to animal cells.

Targeting Gβγ signaling in arterial vascular smooth muscle proliferation: A novel strategy to limit restenosis

Restenosis continues to be a major problem limiting the effectiveness of revascularization procedures. To date, the roles of heterotrimeric G proteins in the triggering of pathological vascular smooth muscle (VSM) cell proliferation have not been elucidated. βγ subunits of heterotrimeric G proteins (Gβγ) are known to activate mitogen-activated protein (MAP) kinases after stimulation of certain G protein-coupled receptors; however, their relevance in VSM mitogenesis in vitro or in vivo is not known. Using adenoviral-mediated transfer of a transgene encoding a peptide inhibitor of Gβγ signaling (βARKct), we evaluated the role of Gβγ in MAP kinase activation and proliferation in response to several mitogens, including serum, in cultured rat VSM cells. Our results include the striking finding that serum-induced proliferation of VSM cells in vitro is mediated largely via Gβγ. Furthermore, we studied the effects of in vivo adenoviral-mediated βARKct gene transfer on VSM intimal hyperplasia in a rat carotid artery restenosis model. Our in vivo results demonstrated that the presence of the βARKct in injured rat carotid arteries significantly reduced VSM intimal hyperplasia by 70%. Thus, Gβγ plays a critical role in physiological VSM proliferation, and targeted Gβγ inhibition represents a novel approach for the treatment of pathological conditions such as restenosis.

The universal ancestor

A genetic annealing model for the universal ancestor of all extant life is presented; the name of the model derives from its resemblance to physical annealing. The scenario pictured starts when “genetic temperatures” were very high, cellular entities (progenotes) were very simple, and information processing systems were inaccurate. Initially, both mutation rate and lateral gene transfer levels were elevated. The latter was pandemic and pervasive to the extent that it, not vertical inheritance, defined the evolutionary dynamic. As increasingly complex and precise biological structures and processes evolved, both the mutation rate and the scope and level of lateral gene transfer, i.e., evolutionary temperature, dropped, and the evolutionary dynamic gradually became that characteristic of modern cells. The various subsystems of the cell “crystallized,” i.e., became refractory to lateral gene transfer, at different stages of “cooling,” with the translation apparatus probably crystallizing first. Organismal lineages, and so organisms as we know them, did not exist at these early stages. The universal phylogenetic tree, therefore, is not an organismal tree at its base but gradually becomes one as its peripheral branchings emerge. The universal ancestor is not a discrete entity. It is, rather, a diverse community of cells that survives and evolves as a biological unit. This communal ancestor has a physical history but not a genealogical one. Over time, this ancestor refined into a smaller number of increasingly complex cell types with the ancestors of the three primary groupings of organisms arising as a result.

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.

Pseudotyping of murine leukemia virus with the envelope glycoproteins of HIV generates a retroviral vector with specificity of infection for CD4-expressing cells

CD4-expressing T cells in lymphoid organs are infected by the primary strains of HIV and represent one of the main sources of virus replication. Gene therapy strategies are being developed that allow the transfer of exogenous genes into CD4+ T lymphocytes whose expression might prevent viral infection or replication. Insights into the mechanisms that govern virus entry into the target cells can be exploited for this purpose. Major determinants of the tropism of infection are the CD4 molecules on the surface of the target cells and the viral envelope glycoproteins at the viral surface. The best characterized and most widely used gene transfer vectors are derived from Moloney murine leukemia virus (MuLV). To generate MuLV-based retroviral gene transfer vector particles with specificity of infection for CD4-expressing cells, we attempted to produce viral pseudotypes, consisting of MuLV capsid particles and the surface (SU) and transmembrane (TM) envelope glycoproteins gp120-SU and gp41-TM of HIV type 1 (HIV-1). Full-length HIV-1 envelope glycoproteins were expressed in the MuLV env-negative packaging cell line TELCeB6. Formation of infectious pseudotype particles was not observed. However, using a truncated variant of the transmembrane protein, lacking sequences of the carboxyl-terminal cytoplasmic domain, pseudotyped retroviruses were generated. Removal of the carboxyl-terminal domain of the transmembrane envelope protein of HIV-1 was therefore absolutely required for the generation of the viral pseudotypes. The virus was shown to infect CD4-expressing cell lines, and infection was prevented by antisera specific for gp120-SU. This retroviral vector should prove useful for the study of HIV infection events mediated by HIV-1 envelope glycoproteins, and for the targeting of CD4+ cells during gene therapy of AIDS.