Avaliação dos efeitos da inibição da maturação nuclear e de antioxidantes sobre a maturação oocitária, fecundação e desenvolvimento embrionário bovino in vitro

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Efeito do estresse subletal com a utilização de LED no desenvolvimento e na qualidade de embriões bovinos produzidos in vitro

Pós-graduação em Medicina Veterinária - FMVZ

Biotecnologias aplicadas à reprodução de éguas velhas

The acceptance of biotechnology for the most equine breeders association had a significant effect in the horse industry, gaining popularity around the world, because the increasing on the genetic gain, allowing the use of sub fertile mares and stallions with high genetics value on reproduction. The embryos in vitro production of human and cattle has been used with success, however in vitro embryo production is not efficient in the horse, as oocyte transfer (OT) and intracytoplasmatic sperm injection (ICSI). The oocyte transfer has been used especially in subfertile old mares presenting reproductive pathologies as: endometrite, cervical and uterine adhesions, blocked oviduct, perineal laceration and ovulation failures. During oocyte recovery process, the oocytes must be collected from immature follicles that need be matured in vitro or in vivo matured oocytes from pre-ovulatory follicles through the transvaginal aspiration guided by ultrasound. The recovered oocyte is transferred to a previously inseminated recipient mare, through the flank laparotomy. The intracytoplasmatic sperm injection (ICSI) is a procedure of in vitro fertilization that needs only one sperm that is aspirated and injected inside the oocyte. The oocytes used, can be from mature and immature follicles. Fresh, cooled and frozen semen can be used, because the procedure not requires a functional sperm. The use of Piezo drill resulted in a breakthrough the pellucid zone, allowing the vibration per minute provided in the sperm injection pipette, a major result of cleaved oocytes, due to a better sperm injection in the oocyte. The embryo transfer can be straight inside the oviduct, as also transcervical transferred after embryo culture produced in vitro. In conclusion both procedures (OT and ICSI) are effective to be used on equine assisted reproduction, getting results even lower than expected, but satisfactory from animal genetically superior

A substituição do meio TCM pelo meio BARC, suplementado com SFB, BSA ou PVA, para maturação de oócitos bovinos in vitro, não incrementa a subsequente produção de blastocistos

This study was carried out to assess the influence of bovine embryo culture medium Beltsville Agriculture Research Center (BARC), supplemented with FCS, BSA or PVA, on the in vitro oocyte maturation, evidenced by cleavage rate and blastocysts production at different developmental stages. Three experiments were performed, as follows: exp.1: addition of FCS to BARC medium at concentrations of 0, 5 and 10%; exp. 2: addition of BSA to BARC medium at concentrations of 0, 4 and 8 mg/ml; exp. 3: addition of PVA to BARC medium at concentrations of 0, 0.5 and 1.0 mg/ml. TCM 199 supplemented with bicarbonate, pyruvate, gentamicin sulfate, FSH, LH and FCS was used as control group. Oocytes obtained from cow ovaries at slaughterhouse were selected in PBS, and then matured in BARC medium supplemented with FSH, LH and gentamicin sulfate, according to the experimental design. Percoll gradient was used for sperm selection and TALP medium for IVF. In vitro embryo culture was in SOF-m medium; a humidified atmosphere with 5% CO2, in air, at 38.7oC was used for all steps. The number of oocytes reaching blastocyst, expanded blastocyst, and hatched blastocyt stages was recorded, respectively at 72 and 168 h post-insemination. ANOVA and Bonferroni t test were used to determine differences among groups. Differences of P<0.05 were taken as significant. Higher percentage (P<0.05) of cleaved oocytes was observed in group TCM + FCS than for the other groups matured in BARC supplemented with FCS or BSA, regardless the concentration used. However, the cleavage rate was similar between groups BARC plus PVA with 1 mg/ml (85.7%) and TCM + FCS (90.8%). Significant difference was found among groups for the production of blastocysts, with the control group yielding a higher number of blastocysts (results ranging from 47.4 to 51.4%, in comparison with groups using BARC + FCS (4.1 to 19.7%), BSA (1.4 to 5.6%) and PVA (5.7 to 10.6%). In conclusion, BARC medium supplemented with different macromolecules did not promote a beneficial effect on in vitro oocyte maturation, resulting in lower rate of cleavage and blastocyst production when compared with TCM + FCS medium.

Efeitos de reguladores do metabolismo lipídico no desenvolvimento in vitro de embriões bovinos e na sobrevivência à vitrificação

Pós-graduação em Medicina Veterinária - FCAV

In Vitro Development and Cell Allocation After Aggregation of Syngeneic Wild Type and Fluorescence-Expressing Bovine Cloned Embryos

Background: The in vitro production (IVP) of embryos by in vitro fertilization or cloning procedures has been known to cause epigenetic changes in the conceptus that in turn are associated with abnormalities in pre- and postnatal development. Handmade cloning (HMC) procedures and the culture of zona-free embryos in individual microwells provide excellent tools for studies in developmental biology, since embryo development and cell allocation patterns can be evaluated under a wide range of embryo reconstruction arrangements and in in vitro embryo culture conditions. As disturbances in embryonic cell allocation after in vitro embryo manipulations and unusual in vivo conditions during the first third of pregnancy appear to be associated with large offspring, embryo aggregation procedures may allow a compensation for epigenetic defects between aggregated embryos or even may influence more favorable cell allocation in embryonic lineages, favoring subsequent development. Thus, the aim of this study was to evaluate in vitro embryo developmental potential and the pattern of cell allocation in blastocysts developed after the aggregation of handmade cloned embryos produced using syngeneic wild type and/or transgenic somatic cells. Materials, Methods & Results: In vitro-matured bovine cumulus-oocyte complexes (COC) were manually bisected after cumulus and zona pellucida removal; then, two enucleated hemi-oocytes were paired and fused with either a wild type (WT) or a GFP-expressing (GFP) fetal skin cell at the 11th and 19th passages, respectively. Following chemical activation, reconstructed cloned embryos and zona-free parthenote embryos were in vitro-cultured in microwells, for 7 days, either individually (1 x 100%) or after the aggregation of two structures (2 x 100%) per microwell, as follows: (G1) one WT cloned embryo; (G2) two aggregated WT embryos; (G3) one GFP cloned embryo; (G4) two aggregated GFP embryos; (G5) aggregation of a WT embryo and a GFP embryo; (G6) one parthenote embryo; or (G7) two aggregated parthenote embryos. Fusion (clones), cleavage (Day 2), and blastocyst (Day 7) rates, and embryonic cell allocation were compared by the. 2 or Fisher tests. Total cell number (TCN) in blastocysts was analyzed by the Student's test (P < 0.05). Fusion and cleavage rates, and cell allocation were similar between groups. On a per WOW basis, development to the blastocyst stage was similar between groups, except for lower rates of development seen in G3. However, when based on number of embryos per group (one or two), blastocyst development was higher in G1 than all other groups, which were similar between one another. Cloned GFP embryos had lower in vitro development to the blastocyst stage than WT embryos, which had more TCN than parthenote or aggregated chimeric WT/GFP embryos. Aggregated GFP embryos had fewer cells than the other embryo groups. Discussion: The in vitro development of GFP cloned embryos was lower than WT embryos, with no effects on cell allocation in resulting blastocysts. Differences in blastocyst rate between groups were likely due to lower GFP-expressing cell viability, as GFP donor cells were at high population cell doublings when used for cloning. On a per embryo basis, embryo aggregation on Day 1 resulted in blastocyst development similar to non-aggregated embryos on Day 7, with no differences in cell proportion between groups. The use of GFP-expressing cells was proven a promising strategy for the study of cell allocation during embryo development, which may assist in the elucidation of mechanisms of abnormalities after in vitro embryo manipulations, leading to the development of improved protocols for the in vitro production (IVP) of bovine embryos.

Algorithms for on-line image registration from multiple views

Images of a scene, static or dynamic, are generally acquired at different epochs from different viewpoints. They potentially gather information about the whole scene and its relative motion with respect to the acquisition device. Data from different (in the spatial or temporal domain) visual sources can be fused together to provide a unique consistent representation of the whole scene, even recovering the third dimension, permitting a more complete understanding of the scene content. Moreover, the pose of the acquisition device can be achieved by estimating the relative motion parameters linking different views, thus providing localization information for automatic guidance purposes. Image registration is based on the use of pattern recognition techniques to match among corresponding parts of different views of the acquired scene. Depending on hypotheses or prior information about the sensor model, the motion model and/or the scene model, this information can be used to estimate global or local geometrical mapping functions between different images or different parts of them. These mapping functions contain relative motion parameters between the scene and the sensor(s) and can be used to integrate accordingly informations coming from the different sources to build a wider or even augmented representation of the scene. Accordingly, for their scene reconstruction and pose estimation capabilities, nowadays image registration techniques from multiple views are increasingly stirring up the interest of the scientific and industrial community. Depending on the applicative domain, accuracy, robustness, and computational payload of the algorithms represent important issues to be addressed and generally a trade-off among them has to be reached. Moreover, on-line performance is desirable in order to guarantee the direct interaction of the vision device with human actors or control systems. This thesis follows a general research approach to cope with these issues, almost independently from the scene content, under the constraint of rigid motions. This approach has been motivated by the portability to very different domains as a very desirable property to achieve. A general image registration approach suitable for on-line applications has been devised and assessed through two challenging case studies in different applicative domains. The first case study regards scene reconstruction through on-line mosaicing of optical microscopy cell images acquired with non automated equipment, while moving manually the microscope holder. By registering the images the field of view of the microscope can be widened, preserving the resolution while reconstructing the whole cell culture and permitting the microscopist to interactively explore the cell culture. In the second case study, the registration of terrestrial satellite images acquired by a camera integral with the satellite is utilized to estimate its three-dimensional orientation from visual data, for automatic guidance purposes. Critical aspects of these applications are emphasized and the choices adopted are motivated accordingly. Results are discussed in view of promising future developments.

Thermoreversible hyaluronan-based hydrogel supports in vitro and ex vivo disc-like differentiation of human mesenchymal stem cells

BACKGROUND CONTEXT The fate of human mesenchymal stem cells (hMSCs) supplied to the degenerating intervertebral disc (IVD) is still not fully understood and can be negatively affected by low oxygen, pH, and glucose concentration of the IVD environment. The hMSC survival and yield upon injection of compromised IVD could be improved by the use of an appropriate carrier and/or by predifferentiation of hMSCs before injection. PURPOSE To optimize hMSC culture conditions in thermoreversible hyaluronan-based hydrogel, hyaluronan-poly(N-isopropylacrylamide) (HA-pNIPAM), to achieve differentiation toward the disc phenotype in vitro, and evaluate whether preconditioning contributes to a better hMSC response ex vivo. STUDY DESIGN In vitro and ex vivo whole-organ culture of hMSCs. METHODS In vitro cultures of hMSCs were conducted in HA-pNIPAM and alginate for 1 week under hypoxia in chondropermissive medium alone and with the supplementation of transforming growth factor β1 or growth and differentiation factor 5 (GDF-5). Ex vivo, hMSCs were either suspended in HA-pNIPAM and directly supplied to the IVDs or predifferentiated with GDF-5 for 1 week in HA-pNIPAM and then supplied to the IVDs. Cell viability was evaluated by Live-Dead assay, and DNA, glycosaminoglycan (GAG), and gene expression profiles were used to assess hMSC differentiation toward the disc phenotype. RESULTS The HA-pNIPAM induced hMSC differentiation toward the disc phenotype more effectively than alginate: in vitro, higher GAG/DNA ratio and higher collagen type II, SOX9, cytokeratin-19, cluster of differentiation 24, and forkhead box protein F1 expressions were found for hMSCs cultured in HA-pNIPAM compared with those cultured in alginate, regardless of the addition of growth factors. Ex vivo, direct combination of HA-pNIPAM with the disc environment induced a stronger disc-like differentiation of hMSCs than predifferentiation of hMSCs followed by their delivery to the discs. CONCLUSIONS Hyaluronan-based thermoreversible hydrogel supports hMSC differentiation toward the disc phenotype without the need for growth factor supplementation in vitro and ex vivo. Further in vivo studies are required to confirm the suitability of this hydrogel as an effective stem cell carrier for the treatment of IVD degeneration.

BENZENE TOXICITY AND THE OCCURRENCE OF BENZENE IN THE AMBIENT AIR OF THE HOUSTON AREA

This study was conducted by either literature review or actual field survey. Results are summarized as follows: (1) Long-term occupational exposure of workers to benzene vapor at levels of 3-7 ppm, 2-3 ppm and 1.6 ppm may result in a decreased level of leucocyte alkaline phosphates, an increased incidence of chromosome aberrations and an increased level of ALA in erythrocytes, respectively; (2) Benzene is capable of causing fetotoxic effects in animals at levels as low as 10 ppm by volume; (3) Exposure of animals to or less than 1 ppm benzene vapor may result in leucopenia, an inverse ratio of muscle antagonist chronaxy and a decreased level of ascorbic acid in fetus's and mother's liver as well as whole embryo; (4) Benzene is causally associated with the increased incidence of pancytopenia, including unicytopenia, bicytopenia and aplastic anemia, and chromosome aberrations in occupational exposure population, and at best benzene must also be considered as a leukemogen; (5) Since it can be emitted into the atmosphere from both man-made and natural sources, benzene in some concentrations is present everywhere in the various compartments of the environment; (6) The findings of the emission of benzene from certain natural sources indicate that reducing benzene to a zero-level of exposure is theoretically impossible; (7) The annual average of benzene concentration detected in the Houston ambient air is 2.50 ppb, which is about 2.4 times higher than the nation-wide annual average exposure level and may have been some health implications to the general public; (8) In the Houston area, stationary sources are more important than mobile sources in contributing to benzene in the ambient air. ^

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.

Expression of mouse telomerase catalytic subunit in embryos and adult tissues

Telomerase is a ribonucleoprotein complex that elongates telomeres, allowing the stable maintenance of chromosomes during multiple cell divisions. Here, we describe the isolation and characterization of the catalytic subunit of mouse telomerase, mTERT (mouse telomerase reverse transcriptase), an essential protein component of the telomerase complex. During embryonic development, mTERT mRNA is abundantly expressed in the whole embryo, especially in regions of intense proliferation. We found that the mTERT mRNA expression in both embryonic and adult tissues is independent of the essential RNA component of telomerase, mTR, and therefore, of the formation of active telomerase complexes. mTERT protein is present exclusively in tissues with telomerase activity, such as testis, spleen, and thymus. mTERT protein is barely detectable in the thymus of mTR−/− mice, suggesting that mTERT protein stability in this tissue may depend on the actual assembly of active telomerase complexes. Finally, we found that mouse and human telomerase catalytic subunit is located in the cell nucleus, and its localization is not regulated during cell cycle progression.

Non-Viral Gene Therapy to Cells of the bovine Intervertebral Disc

Background. Low back pain is an increasing global health problem, which is associated with intervertebral disc (IVD) damage and degeneration. Major changes occur in the nucleus pulposus (NP), with the degradation of the extracellular matrix (ECM).1 Further studies showed that growth factors from transforming growth factor β (TGFβ) and bone morphogenic proteins (BMP) family may induce chondrogenic differentiation of mesenchymal stem cells (MSC).2 Focusing on non-viral gene therapies and their possible translation into the clinics, we investigated if GDF6 (syn. BMP13 or CDMP2) can induce regeneration of degraded NP. We hypothesized that IVD transfected with plasmid over-expressing GDF6 also up-regulates other NP- and chondrogenic cell markers and enhances ECM deposition. Methods. Bovine nucleus pulposus (bNPC) and annulus fibrosus cells (bAFC) were harvested from bovine coccygeal IVD. Primary cells were then electroporized with plasmid GDF6 (Origene, vector RG211366) by optimizing parameters using the Neon Transfection system (Life Technologies, Basel). After transfection, cells were cultured in 2D monolayer or 3D alginate beads for 7, 14 or 21 days. Transfection efficiency of pGDF6 was analyzed by immunohistochemistry and fluorescent microscopy. Cell phenotype was quantified by real-time RT-PCR. To test a non-viral gene therapy applied directly to 3D whole organ culture, coccygeal bovine IVDs were harvested as previously described. Bovine IVDs were transfected by injection of plasmid GDF6 into the center. Electroporation was performed with ECM830 Square Wave Electroporation System (Harvard Apparatus, MA) using 2-needle array electrode or tweezertrodes. 72 h after tranfection discs were fixed and cryosectioned and analyzed by immunofluorescence against GDF6. Results. RT-PCR and immunohistochemistry confirmed up-regulation of GFP and GDF6 in the primary bNPC/bAFC culture. The GFP-tagged GDF6 protein, however, was not visible, possibly due to failure of dimer formation as a result of fusion structure. Organ IVD culture transfection revealed GDF6 positive staining in the center of the disc using 2-needle array electrode. Results from tweezertrodes did not show any GDF6 positive cells. Conclusion. Non-viral transfection is an appealing approach for gene therapy as it fulfills the translational safety aspects of transiency and lacks the toxic effects of viral transduction. We identified novel parameters to successfully transfect primary bovine IVD cells. For transfection of whole IVD explants electroporation parameters need to be further optimized. Acknowledgements. This project was funded by the Lindenhof Foundation (Funds “Research & Teaching”) Project no. 13-02-F. The imaging part of this study was performed with the facility of the Microscopy Imaging Center (MIC), University of Bern. References. Roughly PJ (2004): Spine (Phila), 29:2691-2699 Clarke LE, McConell JC, Sherratt MJ, Derby B, Richardson SM, Hoyland JA (2014), Arthritis Research & Therapy, 16:R67

The role of insulin-like growth factor II and its receptor in mouse preimplantation development

Insulin-like growth factor II (IGF-II) and its receptor, the IGF-II/mannose-6-phosphate (IGF-II/M6P) receptor, are first expressed from the zygotic genome at the two-cell stage of mouse development. However, their role is not clearly defined. Insulin-like growth factor II is believed to mediate growth through the heterologous type 1 IGF and insulin receptors, whereas the IGF-II/M6P receptor is believed to act as a negative regulator of somatic growth by limiting the availability of excess levels of IGF-II. These studies demonstrate that IGF-II does have a role in growth regulation in the early embryo through the IGF-II/M6P receptor. Insulin-like growth factor II stimulated cleavage rate in two-cell embryos in vitro. Moreover, this receptor is required for the glycaemic response of two-cell embryos to IGF-II and for normal progression of early embryos to the blastocyst stage. Improved development of embryos in crowded culture supports the concept of an endogenous embryonic paracrine activity that enhances cell proliferation. These responses indicate that the IGF-II/M6P receptor is functional and likely to participate in such a regulatory circuit. The functional role of IGF-II and its receptor is discussed with reference to regulation of early development.

Oxygen-regulated gene expression in bovine blastocysts

Oxygen concentrations used during in vitro embryo culture can influence embryo development, cell numbers, and gene expression. Here we propose that the preimplantation bovine embryo possesses a molecular mechanism for the detection of, and response to, oxygen, mediated by a family of basic helix-loop-helix transcription factors, the hypoxia-inducible factors (HIFs). Day 5 compacting bovine embryos were cultured under different oxygen tensions (2%, 7%, 20%) and the effect on the expression of oxygen-regulated genes, development, and cell number allocation and HIFalpha protein localization were examined. Bovine in vitro-produced embryos responded to variations in oxygen concentration by altering gene expression. GLUT1 expression was higher following 2% oxygen culture compared with 7% and 20% cultured blastocysts. HIF mRNA expression (HIF1alpha, HIF2alpha) was unaltered by oxygen concentration. HIF2alpha protein was predominantly localized to the nucleus of blastocysts. In contrast, HIF1alpha protein was undetectable at any oxygen concentration or in the presence of the HIF protein stabilizer desferrioxamine (DFO), despite being detectable in cumulus cells following normal maturation conditions, acute anoxic culture, or in the presence of DFO. Oxygen concentration also significantly altered inner cell mass cell proportions at the blastocyst stage. These results suggest that oxygen can influence gene expression in the bovine embryo during postcompaction development and that these effects may be mediated by HIF2alpha.

Blastocyst environment and its influence on offspring cardiovascular health:the heart of the matter

The development of adult-onset diseases such as type II diabetes, obesity and cardiovascular disease is traditionally attributed to adult lifestyle characteristics such as a lack of physical exercise, poor diet and smoking. However, evidence from both human and animal model studies has demonstrated that environmental factors such as an imbalance or reduction in maternal nutrition during gestation can have adverse effects on offspring metabolism and cardiovascular health. The severity and nature of the phenotypic changes induced in offspring is influenced by the period of gestation manipulated. In particular, the mammalian preimplantation embryo in different animal models displays particular sensitivity to environmental factors, either in vivo (maternal diet) or in vitro (embryo culture) that is associated with the onset of cardiovascular dysfunction in adult life. The detailed mechanisms by which environmental conditions can alter postnatal cardiovascular physiology are poorly understood. However, various factors including endothelial function, vascular responsiveness, the renin-angiotensin system, kidney structure and early postnatal growth dynamics have all been recognize as potential contributors. Here, we review the relationship between preimplantation embryo environment and postnatal cardiovascular disease risk, and consider biochemical, molecular, genetic and physiological pathways implicated in this association. © 2009 The Authors Journal compilation © 2009 Anatomical Society of Great Britain and Ireland.