Entwicklung eines pharmakologischen Modells zur Prüfung immuntherapeutischer Antikörper mit dreidimensionalen Tumorzellkulturen in vitro

In den letzten Jahren hat die Tumorbehandlung mit immunologischen Präparaten an Bedeutung gewonnen. Der allgemeine Ablauf der Testung eines Arzneimittelkandidaten sieht vor, zunächst in Zellkulturversuchen und Tierversuchen Wirkweise und Sicherheit, sowie voraussichtliche Abbauwege und mögliche Gefahren so beurteilen zu können, dass sie für einen Einsatz im Menschen in Frage kommen. Zur präklinischen in vitro-Testung werden dabei in der Regel Monolayer-Zellkulturen oder Einzelzellsuspensionen eingesetzt. Der Einsatz von 3D-Zellkulturmodellen, welche den Aufbau von Mikrometastasen oder intervaskuläre Areale in Tumoren exakter widerspiegeln, führt zu wesentlich besseren Voraussagen bezüglich der klinischen Wirksamkeit neuer Präparate. Das Ziel dieser Arbeit war daher die Entwicklung und Anwendung eines neuen 3D-Zellkulturbasierten Systems zur Testung trifunktionaler bispezifischer Antikörper für die Tumorbehandlung, welches sich auch auf andere vergleichbare Präparate übertragen lässt.rnIn meiner Arbeit konnte ich mehrere humane Tumorzelllinien definieren, mit denen es gelang, stabile Co-Kulturen von Multi Cellular Tumour Spheroids (MCTS) mit Peripheral Blood Mononuclear Cells (PBMC) in miniaturisierten Spinner-Flaschen zu etablieren. Spinner-Flaschen, in denen die im Kulturmedium befindlichen Immunzellen, MCTS und Therapeutika ständig frei zirkulieren, sind besonders für eine wirklichkeitsnahe Nachbildung der in vivo-Simulation mit disseminierten Tumorzellen oder mit malignem Aszites geeignet. Diese Art der Kultivierung erlaubte Beobachtungszeiten von ≥20 Tagen für eine große Bandbreite Analysemethoden. Zu den mit dem erstellten Protokoll standardmäßig durchführbaren Analysemethoden zählen unter anderem immunhistochemische Färbungen an Sphäroid-Gefrierschnitten, Vitalitätstest, Untersuchung der Plattierungs-Effizienz, Bestimmung der Sphäroidvolumina, Zytokinbestimmungen aus dem Medienüberstand mit Cytokine Bead Arrays, PCR-Analysen immunzellspezifischer Antigene, sowie durchflusszytometrische Analysen. Diese Methodenkombination erlaubt einen sehr detaillierten Einblick in die Wirkweise und Effizienz neuer Immuntherapeutika aus verschiedensten Blickwinkeln und stellt ein reproduzierbares Testsystem zur präklinischen Testung von Immuntherapeutika dar, das zukünftig als Bindeglied zwischen Monolayer-Zellkulturen und klinischen Prüfungen einen festen Platz einnehmen könnte.rnMit dem beschriebenen 3D-Zellkultur-System wurden in der vorliegenden Arbeit die trifunktionalen bispezifischen Antikörper catumaxomab (unter dem Handelsnamen Removab® für die Behandlung maligner Ascites zugelassen) und ertumaxomab (derzeit in klinischen Prüfungen) hinsichtlich ihrer Wirkweise untersucht. Die Antikörper besitzen im Gegensatz zu herkömmlichen monoklonalen Antikörpern zwei verschiedene Bindungsarme, einer gegen CD3 auf T-Zellen, der zweite gegen EpCAM respektive Her2/neu - beides weit verbreitete Tumorantigene - gerichtet. An ihrem Fc-Teil besitzen sie eine dritte Bindungskapazität, über welche sie an Fcγ RI, -IIa und -III positive akzessorische Zellen binden. Diese Kombination ermöglicht theoretisch die Ausbildung eines Tri-Zell-Komplexes aus T-Zelle, Tumorzelle und akzessorischer Zelle. Dies stellt eine wirkungsvolle Therapieoption unter Ausnutzung der körpereigenen, immunologischen Abwehr dar. rnIm Rahmen dieser Arbeit wurde gezeigt, dass beide Antikörper eine Größenreduktion der Sphäroide mit den entsprechenden Tumorantigenen in gleichem Maße bewirkten und die Plattierungseffizienz durch ertumaxomab dosisabhängig reduziert wurde. Mit dem erstellten Testsystem konnte der Wirkmechanismus von catumaxomab auf Sphäroide der Zelllinie FaDu (Kopf-Hals-Plattenepithelkarzinom) detaillierter gezeigt werden: catumaxomab wirkte dosisabhängig auf die Reduktion der Sphäroidvolumina und die zunehmende Infiltration von CD45+ Zellen, die als T-, NK- und/oder dendritische Zellen identifiziert wurden. Des Weiteren rief die catumaxomab-Gabe eine verstärkte Ausschüttung der Zytokine IL-2, IFN-γ und TNF-α hervor. Diese Ergebnisse sprechen dafür, dass catumaxomab die zelluläre Immunantwort aktiviert.rnDie Standard-Tumorbehandlung beinhaltet die Gabe von Chemotherapeutika. Oft werden dafür Zytostatika mit dem unerwünschten Nebeneffekt auch gesunde proliferierende Zellen anzugreifen verwendet. Dies kann prinzipiell auch die Wirksamkeit der Antikörper-Therapie beeinflussen. Aus diesem Grund wurden in dieser Arbeit zusätzlich vergleichende Kombinations-Versuche mit catumaxomab und einem gängigen Zytostatikum - Cisplatin - durchgeführt. Mit Untersuchungen der Sphäroidvolumina, Vitalitätstests und Plattierungseffizienz konnte gezeigt werden, dass die Wirkung von catumaxomab bei gleichzeitiger Anwendung beider Therapeutika aufrecht erhalten bleibt und diese sogar additiv verstärkt wird. Eine Kombinationstherapie im Menschen ist daher denkbar.rnrn

Therapeutic protein transduction of mammalian cells and mice by nucleic acid-free lentiviral nanoparticles

The straightforward production and dose-controlled administration of protein therapeutics remain major challenges for the biopharmaceutical manufacturing and gene therapy communities. Transgenes linked to HIV-1-derived vpr and pol-based protease cleavage (PC) sequences were co-produced as chimeric fusion proteins in a lentivirus production setting, encapsidated and processed to fusion peptide-free native protein in pseudotyped lentivirions for intracellular delivery and therapeutic action in target cells. Devoid of viral genome sequences, protein-transducing nanoparticles (PTNs) enabled transient and dose-dependent delivery of therapeutic proteins at functional quantities into a variety of mammalian cells in the absence of host chromosome modifications. PTNs delivering Manihot esculenta linamarase into rodent or human, tumor cell lines and spheroids mediated hydrolysis of the innocuous natural prodrug linamarin to cyanide and resulted in efficient cell killing. Following linamarin injection into nude mice, linamarase-transducing nanoparticles impacted solid tumor development through the bystander effect of cyanide.

Self-assembly of sensory neurons into ganglia-like microtissues

Unraveling intra- and inter-cellular signaling networks managing cell-fate control, coordinating complex differentiation regulatory circuits and shaping tissues and organs in living systems remain major challenges in the post-genomic era. Resting on the laurels of past-century monolayer culture technologies, the cell culture community has only recently begun to appreciate the potential of three-dimensional mammalian cell culture systems to reveal the full scope of mechanisms orchestrating the tissue-like cell quorum in space and time. Capitalizing on gravity-enforced self-assembly of monodispersed primary embryonic mouse cells in hanging drops, we designed and characterized a three-dimensional cell culture model for ganglion-like structures. Within 24h, a mixture of mouse embryonic fibroblasts (MEF) and cells, derived from the dorsal root ganglion (DRG) (sensory neurons and Schwann cells) grown in hanging drops, assembled to coherent spherical microtissues characterized by a MEF feeder core and a peripheral layer of DRG-derived cells. In a time-dependent manner, sensory neurons formed a polar ganglion-like cap structure, which coordinated guided axonal outgrowth and innervation of the distal pole of the MEF feeder spheroid. Schwann cells, present in embryonic DRG isolates, tended to align along axonal structures and myelinate them in an in vivo-like manner. Whenever cultivation exceeded 10 days, DRG:MEF-based microtissues disintegrated due to an as yet unknown mechanism. Using a transgenic MEF feeder spheroid, engineered for gaseous acetaldehyde-inducible interferon-beta (ifn-beta) production by cotransduction of retro-/ lenti-viral particles, a short 6-h ifn-beta induction was sufficient to rescue the integrity of DRG:MEF spheroids and enable long-term cultivation of these microtissues. In hanging drops, such microtissues fused to higher-order macrotissue-like structures, which may pave the way for sophisticated bottom-up tissue engineering strategies. DRG:MEF-based artificial micro- and macrotissue design demonstrated accurate key morphological aspects of ganglions and exemplified the potential of self-assembled scaffold-free multicellular micro-/macrotissues to provide new insight into organogenesis.

Increased invasive potential and up-regulation of MMP-2 in MDA-MB-231 breast cancer cells expressing the beta3 integrin subunit

Integrins are a family of transmembrane adhesion receptors that might transduce signals from the extracellular matrix into the inside of cells after ligand binding. In order to investigate whether beta3 integrins expressed in tumor cells might mediate such outside-in signaling, human MDA-MB-231 breast cancer cells that were stably transfected with either beta3 integrin or mock-transfected were investigated in a matrigel degradation assay and a grafting experiment was performed on the developing chicken chorioallantoic membrane (CAM). After cultivation on matrigel for time periods between one and five days, more matrigel was digested in the wells in which beta3 integrin expressing cells were incubated than in wells of mock-transfected cells. Furthermore, extracts of beta3 integrin expressing cells contained higher levels of MMP-2 protein as determined by immunoblotting and more MMP-2 associated gelatinase activity as detected by zymography than extracts of mock-transfected cells. Matrigel degradation and gelatinase activity as well as MMP-2 expression were elevated when beta3 integrin expressing cells were incubated in the presence of the RGD peptide (mimicking an integrin ligand). After grafting on 10 day-old embryonic chicken CAM for three to five days, beta3 integrin expressing cells assembled in spheroids showed higher rates of spreading on the CAM surface and CAM invasion as well as a significant MMP-2 up-regulation compared to mock-transfected cells. The results from the in vivo and in vitro experiments allow the conclusion that the presence of beta3 integrin in MDA-MB-231 breast cancer cells induced an increased MMP-2 expression and activity that might contribute to the enhanced invasive potential observed.

Empirical chemosensitivity testing in a spheroid model of ovarian cancer using a microfluidics-based multiplex platform.

The use of biomarkers to infer drug response in patients is being actively pursued, yet significant challenges with this approach, including the complicated interconnection of pathways, have limited its application. Direct empirical testing of tumor sensitivity would arguably provide a more reliable predictive value, although it has garnered little attention largely due to the technical difficulties associated with this approach. We hypothesize that the application of recently developed microtechnologies, coupled to more complex 3-dimensional cell cultures, could provide a model to address some of these issues. As a proof of concept, we developed a microfluidic device where spheroids of the serous epithelial ovarian cancer cell line TOV112D are entrapped and assayed for their chemoresponse to carboplatin and paclitaxel, two therapeutic agents routinely used for the treatment of ovarian cancer. In order to index the chemoresponse, we analyzed the spatiotemporal evolution of the mortality fraction, as judged by vital dyes and confocal microscopy, within spheroids subjected to different drug concentrations and treatment durations inside the microfluidic device. To reflect microenvironment effects, we tested the effect of exogenous extracellular matrix and serum supplementation during spheroid formation on their chemotherapeutic response. Spheroids displayed augmented chemoresistance in comparison to monolayer culturing. This resistance was further increased by the simultaneous presence of both extracellular matrix and high serum concentration during spheroid formation. Following exposure to chemotherapeutics, cell death profiles were not uniform throughout the spheroid. The highest cell death fraction was found at the center of the spheroid and the lowest at the periphery. Collectively, the results demonstrate the validity of the approach, and provide the basis for further investigation of chemotherapeutic responses in ovarian cancer using microfluidics technology. In the future, such microdevices could provide the framework to assay drug sensitivity in a timeframe suitable for clinical decision making.

TECPR2 Associated Neuroaxonal Dystrophy in Spanish Water Dogs.

Clinical, pathological and genetic examination revealed an as yet uncharacterized juvenile-onset neuroaxonal dystrophy (NAD) in Spanish water dogs. Affected dogs presented with various neurological deficits including gait abnormalities and behavioral deficits. Histopathology demonstrated spheroid formation accentuated in the grey matter of the cerebral hemispheres, the cerebellum, the brain stem and in the sensory pathways of the spinal cord. Iron accumulation was absent. Ultrastructurally spheroids contained predominantly closely packed vesicles with a double-layered membrane, which were characterized as autophagosomes using immunohistochemistry. The family history of the four affected dogs suggested an autosomal recessive inheritance. SNP genotyping showed a single genomic region of extended homozygosity of 4.5 Mb in the four cases on CFA 8. Linkage analysis revealed a maximal parametric LOD score of 2.5 at this region. By whole genome re-sequencing of one affected dog, a perfectly associated, single, non-synonymous coding variant in the canine tectonin beta-propeller repeat-containing protein 2 (TECPR2) gene affecting a highly conserved region was detected (c.4009C>T or p.R1337W). This canine NAD form displays etiologic parallels to an inherited TECPR2 associated type of human hereditary spastic paraparesis (HSP). In contrast to the canine NAD, the spinal cord lesions in most types of human HSP involve the sensory and the motor pathways. Furthermore, the canine NAD form reveals similarities to cases of human NAD defined by widespread spheroid formation without iron accumulation in the basal ganglia. Thus TECPR2 should also be considered as candidate gene for human NAD. Immunohistochemistry and the ultrastructural findings further support the assumption, that TECPR2 regulates autophagosome accumulation in the autophagic pathways. Consequently, this report provides the first genetic characterization of juvenile canine NAD, describes the histopathological features associated with the TECPR2 mutation and provides evidence to emphasize the association between failure of autophagy and neurodegeneration.

Human lung-derived mesenchymal stem cell-conditioned medium exerts in vitro antitumor effects in malignant pleural mesothelioma cell lines.

BACKGROUND The soluble factors secreted by mesenchymal stem cells are thought to either support or inhibit tumor growth. Herein, we investigated whether the human lung-derived mesenchymal stem cell-conditioned medium (hlMSC-CM) exerts antitumor activity in malignant pleural mesothelioma cell lines H28, H2052 and Meso4. METHODS hlMSC-CM was collected from the human lung-derived mesenchymal stem cells. Inhibition of tumor cell growth was based on the reduction of cell viability and inhibition of cell proliferation using the XTT and BrdU assays, respectively. Elimination of tumor spheroids was assessed by the anchorage-independent sphere formation assay. The cytokine profile of hlMSC-CM was determined by a chemiluminescence-based cytokine array. RESULTS Our data showed that hlMSC-CM contains a broad range of soluble factors which include: cytokines, chemokines, hormones, growth and angiogenic factors, matrix metalloproteinases, metalloproteinase inhibitors and cell-cell mediator proteins. The 48- and 72-hour hlMSC-CM treatments of H28, H2052 and Meso4 cell lines elicited significant decreases in cell viability and inhibited cell proliferation. The 72-hour hlMSC-CM incubation of H28 cells completely eliminated the drug-resistant sphere-forming cells, which is more potent than twice the half maximal inhibitory concentration of cisplatin. CONCLUSIONS Our findings indicate that the cell-free hlMSC-CM confers in vitro antitumor activities via soluble factors in the tested mesothelioma cells and, hence, may serve as a therapeutic tool to augment the current treatment strategies in malignant pleural mesothelioma.

Liposome -mediated delivery of all-{\it trans\/}-retinoic acid

Because of its antiproliferative and differentiation-inducing properties, all-trans-retinoic acid (ATRA) has been used as a chemopreventive and therapeutic agent, for treatment various cancers including squamous cell carcinomas (SCCs). Long-term treatment with ATRA is associated with toxic effects in patients leading to acute or chronic hypervitaminosis syndrome. Moreover, prolonged treatment with oral ATRA leads to acquired resistance to the differentiation-inducing effects of the drug. This resistance is attributed to the induction of cytochrome P-450-dependent catabolic enzymes that lead to accelerated ATRA metabolism and decline in circulating levels. Most of these problems could be circumvented by incorporating ATRA in liposomes (L-ATRA) which results in sustained drug release, decrease in drug-associated toxicity, and protection of the drug from metabolism in the host. Liposomes also function as a solubilization matrix enabling lipophilic drugs like ATRA to be aerosolized and delivered directly to target areas in the aerodigestive tract and lungs. Of the 14 formulations tested, the positively-charged liposome, DPPC:SA (9:1, w/w) was found to be most effective in interacting with SCC cell lines. This, L-ATRA formulation was stable in the presence of serum proteins and buffered the toxic effects of the drug against several normal and malignant cell lines. The positive charge attributed by the presence of SA was critical for increased uptake and retention of L-ATRA by SCC cell lines and tumor spheroids. L-ATRA was highly effective in mediating differentiation in normal and transformed epithelial cells. Moreover, liposomal incorporation significantly reduced the rate of ATRA metabolism by cells and isolated liver microsomes. In vivo studies revealed that aerosol delivery is an effective way of administering L-ATRA, in terms of its safety and retention by lung tissue. The drug so delivered, is biologically active and had no toxic effects in mice. From these results, we conclude that liposome-incorporation is an excellent way of delivering ATRA to target tissues. The results obtained may have important clinical implications in treating patients with SCCs of the aerodigestive tract. ^

Description of manganese nodules collected in a metamorphic limestone Cambrian formation near Philipsburg, Montana

In the Philipsburg district, in western Montana a low arch of a Paleozoic limestones has been cut and deformed on the east and a south sides by a small batholith of Tertiary granodiorite. The manganese deposits are confined to an area of about 2 square miles underlain by sedimentary rocks and adjacent to the granodiorite body. The ore, chiefly pyrolusite, was apparently derived from rhodochrosite that was abundant in the veins and had replaced the adjacent limestones. The oxide ore is found chiefly within 600 feet of the surface, though one small body was mined at a depth of 700 feet. Commonly these bodies are aggregates of nodules or spheroids that range in size from that of an egg to that of a coconut or larger. In some places they show an irregular texture somewhat like that of a sponge, and in others the material composing them is loose and friable and apparently structureless. Psilomelane is the principal constituent of many of the nodules, in which it forms layers that alternate with softer oxides.

Desarrollo de estructuras nanofotonicas de campo cercano para intensificacion localizada de luz en celulas solares de banda intermedia - Near-field nanophotonic structures for localized light enhancement in intermediate band solar cells

This doctoral thesis explores some of the possibilities that near-field optics can bring to photovoltaics, and in particular to quantum-dot intermediate band solar cells (QD-IBSCs). Our main focus is the analytical optimization of the electric field distribution produced in the vicinity of single scattering particles, in order to produce the highest possible absorption enhancement in the photovoltaic medium in their surroundings. Near-field scattering structures have also been fabricated in laboratory, allowing the application of the previously studied theoretical concepts to real devices. We start by looking into the electrostatic scattering regime, which is only applicable to sub-wavelength sized particles. In this regime it was found that metallic nano-spheroids can produce absorption enhancements of about two orders of magnitude on the material in their vicinity, due to their strong plasmonic resonance. The frequency of such resonance can be tuned with the shape of the particles, allowing us to match it with the optimal transition energies of the intermediate band material. Since these metallic nanoparticles (MNPs) are to be inserted inside the cell photovoltaic medium, they should be coated by a thin insulating layer to prevent electron-hole recombination at their surface. This analysis is then generalized, using an analytical separation-of-variables method implemented in Mathematica7.0, to compute scattering by spheroids of any size and material. This code allowed the study of the scattering properties of wavelengthsized particles (mesoscopic regime), and it was verified that in this regime dielectric spheroids perform better than metallic. The light intensity scattered from such dielectric spheroids can have more than two orders of magnitude than the incident intensity, and the focal region in front of the particle can be shaped in several ways by changing the particle geometry and/or material. Experimental work was also performed in this PhD to implement in practice the concepts studied in the analysis of sub-wavelength MNPs. A wet-coating method was developed to self-assemble regular arrays of colloidal MNPs on the surface of several materials, such as silicon wafers, amorphous silicon films, gallium arsenide and glass. A series of thermal and chemical tests have been performed showing what treatments the nanoparticles can withstand for their embedment in a photovoltaic medium. MNPs arrays are then inserted in an amorphous silicon medium to study the effect of their plasmonic near-field enhancement on the absorption spectrum of the material. The self-assembled arrays of MNPs constructed in these experiments inspired a new strategy for fabricating IBSCs using colloidal quantum dots (CQDs). Such CQDs can be deposited in self-assembled monolayers, using procedures similar to those developed for the patterning of colloidal MNPs. The use of CQDs to form the intermediate band presents several important practical and physical advantages relative to the conventional dots epitaxially grown by the Stranski-Krastanov method. Besides, this provides a fast and inexpensive method for patterning binary arrays of QDs and MNPs, envisioned in the theoretical part of this thesis, in which the MNPs act as antennas focusing the light in the QDs and therefore boosting their absorption

Sobre la Arquitectura en la definición del Paisaje

Comúnmente la Arquitectura se manifiesta en los edificios como un hecho de la realidad que tiene siempre carácter contemporáneo y ése debe ser el valor supremo de un arte que no distingue entre antiguo y moderno, por afectar al presente y ser contemporáneo como experiencia. Los objetos se insertan irremediablemente en su medio e incluso llegan a definirlo en ocasiones: así, arquitectura y paisaje, aparecerán a veces confundidos con diferencias difíciles de determinar. El término “paisaje” es relativamente moderno y se deriva de ciertas representaciones gráficas o pictóricas producidas en Occidente en época algo posterior al Renacimiento. No obstante, el hecho de que una palabra se pueda escribir o se cite no quiere decir que la realidad a la que responde no pueda existir, pues lo escrito es solamente un medio de expresión, y es obvio que existen otros quizá más antiguos y de idéntica importancia, pues la propia escritura no es más que el reflejo de un fenómeno que ya se ha producido con anterioridad. Parece así que el testimonio de la realidad viene dado por distintos contextos en lo que suele llamarse “cultura” de modo que las culturas pueden tener aspectos de gran interés mediante diferentes sistemas de expresión variados sin que eso tenga que pasar forzosamente por el filtro de un alfabeto y una escritura. A tenor de los primeros descubrimientos, parece que la cuestión de escribir tuvo originalmente un carácter de constatación de apunte contable y tampoco se puede establecer con certeza si algunos utilizaron la escritura como el so-porte adecuado para referir mitos o historias, pues la Arqueología no ha proporcionado sino testimonios fragmentarios; de lo que si está repleta sin duda la historia es de apuntes contables. Otra cuestión que suscita dudas es la propia definición de escritura, pues parece que los más antiguos modos de expresión escrita se reducen a pictogramas que resultan aún indescifrables. Lenguas y toponimia son también herramientas muy útiles para bucear en el pasado, pero aún así persisten dudas en las escrituras que pro-vienen de las representaciones primordiales: la propia escritura en sus orígenes parece alimentarse de las propias intuiciones inmediatas de la representación, algo que evolucionaría representando esa realidad urgente e inmediata del control o el abastecimiento que luego se convertirían en los fragmentos de texto que han podido conservarse. Es notorio, sin embargo, que ese conjunto enorme de convenios gráficos estaba sujeto a la palabra y la desaparición de las culturas determinó también la desaparición consecuente de sus lenguas, y esos signos - fueran o no escritura - han pasado definitivamente a las zonas oscuras del conocimiento. Escribir supone también disponer de capacidad de abstracción gráfica que diferencie la palabra y la cosa, y es posible que la razón que llevara a ese descubrimiento fuera simplemente el de una economía de signos, pues escritura y lenguaje son limitados por definición, mientras que las cosas del mundo son innumerables: ningún idioma contiene todas las palabras para describir todo lo que puede aparecer. Aparentemente, ese el motivo por el cual existe un localismo – un término específico que refiere algo que existe en un lugar y no en otros - en lo que se refiere a dar el nombre a la cosa y también fuente de paradigma entre los pueblos que se consideran primitivos y los que se consideran civilizados. Debe tenerse en cuenta también que esa transposición se da en ambos sentidos, de modo que las culturas más aisladas también incorporan conceptos que carecen de una explicación racional consistente. Las mitologías son así eternas, pues sirven para explicar realidades que carecen de un entendimiento alcanzable y está también bastante claro que los lenguajes resultan ser un enigma, como realidad autónoma que queda en-cerrada y se explica en sí misma. Parece también que los primeros en escribir las consonantes aisladas fueron los pueblos semíticos occidentales de las orillas del Mediterráneo, pueblos que crearon un alfabeto silábico que llegó a ser utilizado incluso por los tartesios en el sur de la península ibérica como el primer alfabeto de toda Europa occidental. Resulta patente que el término “paisaje” se ha nutrido siempre de representaciones, bien escritas o artísticas, pero queda también claro que esas representaciones se suponen derivadas de la creencia en una idea de paisaje como algo que se ve representado en sí mismo, es decir, como la imagen de una realidad externa situada fuera del individuo. Eso es un hecho trascendente, pues el paisaje requiere lejanía de la cosa, de modo que el actor – aún sabiéndose inserto en su paisaje – es incapaz de percibirlo desde dentro. En el paisaje ocurre igual que en un teatro o en una representación: los actores son conscientes de su papel y su posible cometido, pero no son los que realmente pueden disfrutar de la eficacia o de la propia presencia de la obra, pues forman parte de ella. La idea de paisaje proviene de una lectura externa a la de los propios protagonistas del drama, de modo que si quieren ser un crítico del paisaje se debe abandonar la representación para mirar el espectáculo desde una distancia adecuada, al fin y a cabo externa. De ese modo, aparece la primera constatación del hecho del paisaje como una realidad construida por objetos y personajes, pero, sobre todo, es una realidad construida por miradas. Llama también la atención otorgada en las lecturas de los especialistas hacia esa referencia externa - artística si se quiere - sobre el término “paisaje”, de modo que la bibliografía no especializada sobre el particular siempre acaba en tratados de pintura o poesía. Parece sin embargo que el hombre y sus paisajes son realidades indisolubles desde la propia aparición de la especie, de modo que llevar la cuestión hacia términos exclusivamente esteticistas parece una posición parcial. Hombre y medio han formado siempre una sola unidad, aunque esa unidad se haya convertido en determinados casos en sinónimo de depredación y destrozo. Sin embargo, esa destrucción crea igualmente un paisaje como desolación que es resultado del propio quehacer del hombre: elementos que también poseen un contenido sustancial de memoria en los paisajes industriales como un momento de la Historia previo a la automatización y a la globalización de la vida actuales. Quizá el concepto más interesante desde el punto de vista teórico sea precisamente el de esa cualidad del paisaje como mirada, algo externo producido por el contemplador en un momento ajeno a la pertenencia, una mirada que no es tan sólo estética sino comprensiva, gozosa, activa o analítica - pues existen tantas maneras de mirar como sujetos - sin que pueda definirse con precisión esa contemplación más que en términos que alcanzan la propia individualidad. En términos poéticos, también podría establecerse como ese conjunto de miradas individuales crean también una estructura que hace que ese paisaje se valore y se comprenda, de modo que desde ese punto de vista ese paisaje supone una creación colectiva. Con respeto o como depredador, el hombre se ha instalado en su medio, y al hacerlo, ha dejado huellas dentro del propio paisaje que hacen que tome una determinada significación. Naturalmente, existe una teoría que distingue entre “país” y “paisaje”, asumiendo para el primero la noción exclusiva de territorio en la cual el hombre no tiene papel estético alguno. He intentado muchas veces comprender esa posición, sin acabar de entender el planteamiento que la sustenta: parece que la visión de la cosa estará siempre humanizada, aún en las zonas “vírgenes” o intactas, pues la propia visión hace que el objeto se modifique en su propia unidad perceptiva, creando una curiosa indeterminación que lleva a los conocidos equívocos entre imagen real y representación. Efectivamente, la visión de la cosa requiere de una cultura y unos medios que la informan, de modo que un texto, una pintura, una descripción o una fotografía quedarán ya humanizadas por el propio hecho de ser informadas, pues ello otorga una forma a priori. De ese modo, el paisaje figura inscrito en una función que establece tanto aspectos de un paisaje posible como aspectos del paisaje descrito y solamente podrá hablarse sobre la potencialidad del estado final de ese paisaje y nada del propio paisaje en sí, mientras ese paisaje no quede humanizado por el propio observador. Esta cuestión obliga a elegir una definición de paisaje alejada de presupuestos esteticistas para acercarla a los puramente culturales, de modo que no existe otra vía para la investigación que contemplar esa realidad física en función de las coordenadas del hombre que la habita, dotada del contenido correspondiente a esa revelación. Lejos de las posiciones de la geomorfología, el término “paisaje” implicará así unas condiciones determinadas de contemplación por parte de un sujeto por el cual el espectáculo queda humanizado en dicho acto.Cabe pensar también si no es cierto que todos los paisajes requieren de esa condición humanizada, pues aunque no estén habitados están siempre ocupados por esa mirada que los habita, al igual que un escenario sin público que carece de vigencia. A partir de ahí se sitúan las coordenadas en donde este trabajo configura la presencia de la arquitectura en la definición del paisaje, una premisa que parece ya venir otorgada desde el principio, pues esa misma mirada del espectador ya está dotando de un sentido de orden y jerarquía a la contemplación, unas cualidades que están en la base de toda actividad arquitectónica, De hecho la propia definición de “monumento natural” - en si misma una contradicción – expresa ese conflicto, dotado de un fenómeno de admiración y solape entre cultura y naturaleza, como conceptos enfrentados desde su origen. La conclusión sobre el dilema propuesta en la tesis no ha sido otra que suponer que esas dos realidades que son la cultura y el paisaje se han solapado desde el principio de los tiempos del hombre formando un binomio indeslindable. Se ha dicho antes que el proceso de invasión del territorio por el hombre es significativo, y esa significación es la que origina una creación autónoma que se aísla como un concepto abstracto de los entes naturales, tomándolos como material de trabajo, y estableciendo una oposición conceptual en la realidad perforada e interpretada por el hombre que viene a oponerse a lo que supone la caja cerrada e ignota del enigma del mundo. La cuestión de la significación del hombre sobre el territorio se subsume en la creación de unos productos que permanecen y que son testimonio de la propia cultura, de forma que la cantidad de rastro que el hombre deja sobre el territorio contribuye en mayor medida a la cualificación del paisaje como concepto. Eso lleva a establecer que cualquier paisaje - por el mero hecho de serlo y ser definido así – es ya cultural, puesto que está configurado por los productos de la cultura. Las palabras que puedan quedar encerradas en las piedras de los monumentos o de las ciudades son las de los hombres que trabajaron allí, y también las de los que las habitaron: más aún, el propio sentido del paisaje y su conservación vienen determinados por la presencia del hombre como único interprete de conceptos como ecología o conservación, algo que se pone de manifiesto también en la espantosa devastación que producen los fenómenos propios de la naturaleza. La historia natural, al igual que la vida, están conformadas por éxito y devastación, sin que uno y otra tengan especial preferencia, pues la preferencia se alimenta de otra clase de conceptos. La cuestión de atribuir valores morales al mundo natural es algo muy antiguo, y quizá sea la fuente o el manantial de las primeras religiones, una cuestión que se une a la indefectible noción de mortalidad que define la existencia del hombre frente a la inmanencia de la naturaleza. Esa propia naturaleza está dotada intuitivamente de un carácter “inocente” suponiendo que la inocencia es lo opuesto a la sabiduría. La cuestión es bien otra, ya que la naturaleza no posee ni siquiera algo que pueda definirse como “carácter”, en el sentido personal del término. La cuestión no cae, evidentemente, del lado de las definiciones o de las cualidades, sino del propio análisis de la realidad que el hombre va construyendo, destruyendo, embelleciendo o perjudicando para definir el territorio que habita, interponiendo a su manera recursos, instalaciones y en definitiva todos los testimonios posibles como principal manifestación de su esencia. Entre los artefactos que el hombre produce, uno de los más persistentes y más conspicuamente descritos es el de la arquitectura - entendida aquí en un sentido amplio - es decir, como el conjunto de modificaciones del espacio y del territorio. El espacio se puede modificar de muchos modos, pero en cualquiera de los casos constituye una de las huellas más características que el hombre establece como manifestación física de su propio ser discursivo. También la arquitectura ha demostrado ser una de los fenómenos más persistentes, incluso más que la propia lengua que la origina en su discurso primero antes que pueda ac-cederse a una idea sobre la conformación del material. Es paradigmático que el episodio descrito sobre la Torre de Babel en la Biblia la cuestión de la ambición de los hombres frente a Dios, representada precisamente en una arquitectura, se asimile a la discusión sobre el lenguaje primordial. La cuestión no es baladí, pues el fenómeno de la creación es algo que se concede exclusivamente a los dioses, que por esa razón habitan los territorios a los que los hombres no pueden llegar; territorios de albergue en los que las mitologías sitúan a dioses y demonios, con amplios espacios intermedios donde situar a las divinidades menores, héroes o seres híbridos en los que la coyunda de los dioses con los humanos produce sujetos que alivian el sentido de la mortalidad. El comentario del Génesis también concede un valor a la técnica, al mito de Prometeo y al fuego como creador de excelencia. Frente al progreso prometeico, se postula el valor divino, y la única forma posible de paliar ese progreso es la confusión del lenguaje, pues eso será lo que produzca la dispersión y la falta de acuerdo. La cuestión también puede explicar esa afición tan común por lo canónico en arquitectura que se mantiene a lo largo de grandes períodos, al igual que una gran máquina de inercia. Parece que los conceptos primordiales de la arquitectura basados en elementos simples como el hito, el dintel, lo recto y lo curvo, la rampa o la escalera, y el uso distinto o cualificado que se otorga a la piedra, el ladrillo, la madera o el metal como componentes primordiales del material arquitectónico se haya mantenido a lo largo de muchos milenios sin apenas cambios, al igual que ocurre con las costumbres alimenticias que siguen una progresión ascendente a través de lo crudo, lo asado y lo cocido, hasta obtener distintos grados de refina-miento, pero que siempre se sustentan en la sensación primigenia. El descubrimiento de la arquitectura proviene de un cierto manejo de las dimensiones, y consiguientemente de la geometría. Pero la geometría es cosa abstracta al igual que el lenguaje, de modo que para poder realizar arquitectura se hace necesaria esa capacidad de abstracción primera que luego permite la realización de un dispositivo. La realidad y su número exhiben un divorcio, al igual que las cosas y sus nombres exhiben el suyo, y el análisis numérico es solamente una forma de ver la realidad, una forma rigurosa y exacta – aunque parcial - que solamente representa el modelo ideal al cual la realidad se aproxima en mayor o menor medida: esa aproximación matemática hace que el universo pueda condensarse parcialmente en números, al igual que la realidad puede condensarse en nombres, pero ni el nombre ni el número reflejarán el mundo en toda su complejidad. El número es quizá solamente un medio de describir las cosas, como lo serían las formas puras que responden a una configuración matemática que pueda producirse en teoría en cualquier parte del Universo. Sin embargo, para el ejercicio de la arquitectura es preciso acudir a esa simplificación que exige la visión abstracta del plano como una sección cierta realidad como un corte abstracto del elemento considerado. Con su traza o sin ella, con la propia expresión matemática que lo describa o sin precisarla, esa intuición del plano como elemento generador del espacio es anterior a aquella expresión, al igual que el habla fue anterior a la escritura, pues solamente se produjo a través de ella y como refuerzo y sustituto de la memoria. Existen así abstracciones de la memoria que aparecen derivadas de los signos de la naturaleza aparecidos solamente de forma eventual y fragmentaría: así ocurre con la línea, el cuadrado y el círculo, formas iniciales y abstractas sonsacadas en cierta medida de la observación que dan origen a los primeros signos de la arquitectura primordial, de modo que cuadrados y círculos, elevados a prismas y superficies esféricas o esferoides aparecen en tumbas y edificios elementales de los primeros tiempos mediante una geometría primordial que se superpone al paisaje en el que se inserta. Es cierto también que esas formas se encuentran ya aproximadas en objetos que se encuentran en el medio físico, líneas en extremidades, ramas y miembros; ángulos rectos en algunos cristales que se observan mediante corte o formas circulares en astros y cráteres, pero esa realidad solamente presenta su forma aproximada y no su abstracción pura, de igual modo que el jeroglífico propondrá al ave rapaz para representar la idea de vigilancia y la buena vista, o incluso que la imagen del propio ojo sustituya la idea del Dios que todo lo ve en las culturas anti-guas. La elección fue resuelta, después de muchos intentos y aproximaciones, con la adopción del ángulo recto como un artificio fácil para el replanteo a través del triángulo sagrado 3:4:5 que ya se utilizó para construir las pirámides de Egipto, y que dio origen también a la mayor parte del urbanismo antiguo, coexistiendo con la forma circular primordial en el tipo denominado “tholo”. Aquella trama cuadrangular era uno de los patrones de relleno del espacio más comunes y compactos, y esa fue probablemente la razón por la que en tiempos muy posteriores fuera adoptada como una forma eficaz permanente de organización frente al desorden topológico que procura el conjunto de asociación de plantas circulares. Otra cuestión paradigmática es que esos conceptos primordiales e ignotos - que convergen en el mismo origen de las civilizaciones - se conviertan luego en algo canónico, a través del uso. El canon en sí mismo es algo ideal, como una norma aplicable a objetos de una realidad que ha sido creada solamente como indicio del ca-non, algo abstracto que tiene proporciones estrictas que son siempre las mismas y no obedece a criterios racionales: será absurdo sin embargo buscar el canon griego entre los templos de época como algunos lo hicieron, pues los edificios solamente se aproximan a los ejemplos canónicos y por esa razón se habla del “dórico del Partenón” que es diferente del de Egina o del de Paestum, siendo todos ellos evidentemente dóricos. Sin embargo, esa idea resulta útil al tratadista, al teórico de la arquitectura y al historiador, aun-que solamente refleje una vaga idea de lo que sucede más allá del tratado. Otra cuestión es la sutileza de los ejemplos de arquitectura, y del mismo modo que los cuerpos de los seres vivos jamás son simétricos, aunque respondan a un diseño simétrico general derivado de las condiciones de la división celular, los edificios supuestamente canónicos en los que los especialistas se inspiraron para definir los órdenes clásicos no disponen de esa simetría modular exacta, sino que adaptan un modelo general al lugar siempre cambiante del emplazamiento, es decir, se adaptan con habilidad a la vez que configuran el paisaje en el que se insertan. La luz de los distintos intercolumnios del Partenón es ligeramente distinta, aunque guarde un evidente sentido de simetría axial, de manera que aquellos “órdenes” que formaron la Teoría de la Arquitectura no son más que una bella interpretación sectorial de la arquitectura antigua elaborada por los tratadistas del Renacimiento y, posteriormente, por los neoclásicos. Parece, sin embargo, que ese ansia por el canon como claridad de lenguaje es algo consustancial al desarrollo de la arquitectura, una lingua franca que tiende a evitar la dispersión producida entre los mortales por los dioses antiguos, de modo que si no era posible llegar al cielo de los dioses se procuró que el lenguaje de la Tierra fuera al menos inteligible para poder entenderse entre los hombres. Parece que la estructura del poder siempre requirió de un determinado nivel de organización, y también que las instrucciones se entendieran con claridad: parece que en nuestros tiempos esos antiguos cánones se han sustituido por la obediencia a normas abstractas, dictadas por instituciones también algo abstractas y que tienen nombres divertidos compuestos por siglas, aunque no se conozca bien su virtud. El canon actual está constituido por “normas” que dejan tranquilos a algunos, aunque parece quizá que todo ese entramado formal que sirve como anestesia para el cuerpo social procura también la destrucción de los bosques en formas de montañas ingentes de papel burocrático. Durante muchos siglos las normas fueron otras, en la forma de un canon del cual nadie podía escapar: aún así, mediante la utilización de cánones o sin ellos, la arquitectura prosperó en la civilización desde los primeros refugios cavernarios o los abrigos primigenios y de ese modo fue configurando la realidad, es decir, el paisaje. Como antes se dijo, ese es un viaje de ida y vuelta en el cual ambos se confundían y subsumían: el manejo de las formas y lenguajes de la arquitectura posibilitaría con el tiempo la distinción entre el campo en donde reina el caos y la ciudad, en donde reina teóricamente el orden, creando un divorcio que duraría milenios y que aún persiste. Esa oposición generaría también una forma de paisaje y una serie de usos simbólicos y sagrados de los elementos de la arquitectura - como son puertas y murallas - que se han mantenido conceptualmente aunque hoy las ciudades ya no posean murallas físicas, ni puertas que se cierran con la llegada de la noche. En ese sentido, la arquitectura ha podido definir el paisaje, entendiendo la arquitectura no solamente como los edificios en sí, sino como el hecho de la manifestación del hombre sobre el territorio, de modo que no podrá hablarse de paisaje sin su arquitectura, del mismo modo que no puede hablarse de paisaje sin hombres. Por esta razón, la Tesis habla sobre Arquitectura y Paisaje, pero más particularmente sobre el hecho de la arquitectura en la definición del paisaje, es decir, de como los hechos arquitectónicos van a determinar o no la cualidad y la calificación de un paisaje. Deberá partirse en primer lugar de una definición de lo que se entiende y se ha entendido comúnmente por paisaje: igualmente, y habida cuenta del contexto en el que sitúa el propio trabajo de tesis, la cuestión solamente se circunscribirá a lo que se entiende como cultura occidental, pues el desarrollo de la civilización presenta siempre un color local que hace que el análisis de un fenómeno de esta envergadura tenga distintas connotaciones en las distintas áreas de civilización. De igual modo, y habida cuenta también que el paisaje se construye a través de todas las manifestaciones culturales, se hará a veces necesario indagar en otras disciplinas no arquitectónicas para comprender el alcance de algunos casos en los cuales los restos arquitectónicos han desaparecido, o en los que subsisten escasas trazas. Una definición tan amplia de la Arquitectura llevaría a escribir un relato sobre toda la cultura occidental y por ese motivo solamente se han esbozado ideas sobre la aparición de esos signos sobre el paisaje basados en elementos antiguos y primigenios que se repiten con insistencia y van dando lugar al variado repertorio de la arquitectura occidental cómo conformación de ideas sobre el me-dio y sobre el mundo que se percibe y se habita. ABSTRACT About Architecture in defining Landscape. Abstract Architecture is commonly manifested in buildings as a fact of reality that has always a contemporary character and that should be the highest value of an art that does not distinguish between ancient and modern, to affect the present and be contemporary as experience. Objects are inserted irremediably in their midst and even sometimes come to define it: thus, architecture and landscape, appear sometimes confused with differences difficult to determine. However, the term "landscape" is relatively modern and is derived from certain graphical or pictorial representations produced in the West in some subsequent period to the Renaissance. The fact that a word can be written or quoting does not mean that the reality that can not be answered, because the writing is only a mean of expression, and it is obvious that there are other and perhaps older equally important, because the writing itself is nothing more than the reflection of a phenomenon that has occurred before. It thus appears that the testimony of reality is given by different contexts in what is called "culture", so that cultures can have aspects of great interest by many different expression systems without this necessarily have to pass through the filter of alphabet and writing. Under the initial findings, it appears that the question of writing originally had a character and finding accounting entries, and it can not be established with certainty whether some used writing as the support appropriate to refer myths or stories, because archaeology has provided only fragmentary evidence. Another issue that raises questions is what can properly be defined as writing, it seems that the oldest modes are reduced to writing pictograms are still indecipherable. Languages and place names are also very useful tools for diving in the past, but still questions remain in the scriptures that come from the primordial representations: either it is very well defined what the own writing in its origins: the beginnings seem to feed on immediate intuitions of representation, which would evolve representing reality that urgent and immediate control or supply which is then inherited into fragments. It is noteworthy, however, that this huge set of graphics agreements was subject to the word and the disappearance of cultures determined also the consequent disappearance of their languages, and those signs - whether or not they write - have passed definitively to dark areas of knowledge. Writings supposed also the capacity of abstraction graph differentiates the word and the thing, and it is possible that the reason to carry this discovery was simply that of an economy of signs, for writing and language are limited by definition, while the things of the world are innumerable: no language contains all words to describe all that may appear. Apparently, that's why there is a localism - a specific term that refers to something that exists in one place and not in others - in regards to name the thing and also the source of paradigm among peoples are considered primitive and civilized. It should be noted also that transposition occurs in both directions, so that the most isolated cultures also incorporate concepts that lack a consistent rational explanation. Mythologies are eternal and therefore serve to explain realities that lack an understanding achievable and also pretty clear that languages happen to be an enigma, as an autonomous reality that is enclosed and explains itself. It also seems that the first to write consonants were isolated western Semitic peoples from the shores of the Mediterranean, peoples who created a syllabic alphabet came to be used even by tartesios in southern Iberia as the first alphabet in Western Europe. It is clear that the term "landscape" has always nurtured representations, either written or artis-tic, but it is also clear that these representations are assumed arising from belief in an idea of landscape as something that is represented in itself, as the image of a reality external located outside the individual. That is an important fact because the landscape requires remoteness of the thing, so that the actor - even knowing insert in landscape - is unable to perceive from within. The landscape is just as in a theatre or a performance: the actors are aware of their role and their possible role, but they are not the ones who can really enjoy the efficiency or the presence of the work itself, as part of it. The idea comes from an external landscape reading the principles of players in the drama, so if you want to be a critic of the landscape should leave the actual representation to watch the spectacle from a safe distance, finally external. Thus, the first finding of fact of the landscape appears as a reality constructed by objects and characters, but above all, a reality constructed by looks. Also noteworthy given the readings of specialists to the external reference - art if it could be - on the term "landscape", so no specialized literature on the subject always ends in treatises on painting or poetry. It seems however that the man and his landscapes are inseparable realities from the very onset of the species, so bring the issue into terms exclusively aesthetics seems a partial position. Man and environment have always been a single unit, but that unit has become synonymous with certain cases predation and destruction. Nevertheless, this destruction also creates a landscape as desolation that results from proper task of man elements that also have substantial contents of memory in industrial landscapes as a time of pre-automation history and globalization of current life. Perhaps the most interesting from a theoretical point of view is precisely that quality of landscape as something external produced by the viewer in a strange time of membership, a look that is not only aesthetic but sympathetic, joyful, active concept or analytical - because there are so many ways to look as subjects - it may not be precisely defined that contemplation rather than in terms that reach one's individuality. In poetic terms, it could also be set as the set of individual gazes also create a structure that makes this landscape is valued and understood, so from that point of view that landscape is a collective creation. With respect or as a predator, man has settled in his environment and in doing so has left traces in the landscape itself that make take a certain significance. Naturally, there is a theory that distinguishes what is "home" and what is "nature" providing for the first notion of the exclusive territory in which man has no aesthetic role. I tried many times to understand this position, without understanding the approach that supports: it seems that the vision of the thing is always humane, even in the "virgin" or untouched areas, as the vision itself makes the object modified in its own perceptual unit, creating a curious indeterminacy leading to the known misunderstandings between real image and representation. Indeed, the vision of the thing requires a culture and means that the report, so that a text, a picture, a description or photograph will be humanized by the very fact of being informed, as this provides a way a priori. Thus, the landscape provides a function that sets both aspects of a potential landscape as described aspects of the landscape and can only talk about the potential of the final state of the landscape, while the landscape remains humanized by the observer himself. This question forces to choose a definition of remote landscape budgets purely cultural, so there is another way for research to contemplate that physical reality in terms of the coordinates of the man who inhabits gifted content corresponding to that revelation. Far from the positions of the geomorphology, the term "landscape" and involve a certain condition by contemplation of a subject for which the show is humanized in the act. It should also consider, in the light of the above, if it is not true that all landscapes require that humanized condition, because although they are not inhabited they are always occupied by the look that dwells, like a stage without audience produces no consistent effect. From there the coordinates where this work sets the presence of architecture in defining landscape, a premise which seems to come from the beginning given lie, because that same look is already giving the viewer a sense of order and hierarchy to contemplation, qualities that are at the basis of all architectural activity, in fact the very definition of "natural monument" - in itself a contradiction - expresses this conflict, which has a phenomenon of admiration and overlap between culture and nature as concepts faced since its inception. The conclusion on the dilemma proposed in the thesis has not been another to assume that these two realities are the culture and landscape have overlapped since the beginning of man time forming a pair. It has been said before that the process of invasion of the territory by man is significant, and that meaning is the originating autonomous creation that is isolated as an abstract concept of nature, taking them as working material, and establishing a conceptual opposition in reality and punched by the man who comes to oppose representing the closed and unknown to the enigma of the world interpreted. The question of the significance of the man on the land is subsumed in the creation of products that remain and are testimony of their own culture, so that the trace amount that the man leaves the territory contributes most to the qualification landscape as a concept. That brought into any landscape - by the mere fact of being and being well defined - is already cultural, as it is configured by culture products. The words that can be locked in the stones of the monuments or cities are those of the men who worked there, and also of those who inhabited: even more, the sense of the landscape itself and its conservation are determined by the presence of man as the sole interpreter of concepts such as ecology or conservation, something which becomes manifest also in the awful devastation that produce the phenomena of nature. The natural history, like life, are shaped by success and devastation without special preference, the preference is used for feeding on other kinds of concepts. The question of moral values attributed to the natural world is very ancient, and may be the source or the source of the first religions, an issue that joins the unfailing notion of mortality that defines the existence of man against immanence of nature. That nature is endowed intuitively an "innocent" character assuming that innocence is the opposite of wisdom. The other issue is well, since nature does not even have what is defined as "character" because that is something that serves to qualify beings, but not objects. The question does not fall clearly on the side of the definitions or qualities, but from the analysis of the reality that man is building, destroying or damaging to define the territory, interposing his way resources facilities and possible witness as the main manifestation of its essence. Among the artifacts that man produces one of the most persistent and most conspicuously disclosed is architecture as a way of modification of space and territory. The space can be modified in many ways, but in either case is one of the most characteristic traces that man establishes as a physical manifestation of his own discourse being. Architecture has also proven to be one of the most persistent phenomena, even more than their own language that originates in his speech first. The paradigm wrote in the episode described on the Tower of Babel in the Bible shows the question of ambition of men before God - represented precisely in architecture - is assimilated to the discussion about the primary language. The issue is not trivial, since the phenomenon of creation is something that is granted exclusively to the gods, for that reason inhabit the territories to which men can not reach; territories where the hostel located mythologies gods and demons, with large gaps where to place the minor deities, heroes or hybrid beings where the yoke of the gods with human subjects produces relieving sense of mortality. The commentary on Genesis also gives a value to the art, the myth of Prometheus and fire as creator of excellence. In front of promethean progress, divine value is postulated, and the only possible way to alleviate this progress is the confusion of language, because that is what will produce the dispersion and lack of agreement. The issue may also explain such a common passion for the canonical architecture maintained throughout long periods, like a great machine inertia. It seems that the main concepts of architecture based on simple elements like milestone, lintels, straight and curved stairs or ramps, or other qualified and used are granted to the stone, brick, wood or metal as the primary components of the architectural material maintained throughout many millennia are almost unchanged, as is the case with the eating habits that follow a progression through the raw, the cooked and roasted or fried until obtain different degrees of refinement, but always are based on the primal feeling. The discovery of the architecture comes from a management dimensions, and consequently the geometry. But the geometry is abstract thing like the language so that to make architecture that first absorption capacity which then allows the realization of a device is necessary. Reality and its number exhibit a divorce, like things and their names displayed his; numerical analysis is just one way of seeing reality, rigorous and accurate - though partial - only represents the ideal model to which reality is coming to a greater or lesser extent: the mathematical approach makes the universe can condense on numbers, like reality may condense on names, but neither the name nor the number will reflect in all its complexity. The number is only a means of describing things, such as the pure forms that match setup a mathematical theory that occurs anywhere in the universe. However, for the practice of architecture is necessary to go to the simplification that requires abstract view of a section plane of reality, as an abstract element considered cutting. With its trace or not, with the mathematical expression that describes what or without clarify, this intuition of the plane as a generator of space predates his own expression, like speech preceded the writing, which only occurred through it and as reinforcement and replacement of memory. There are abstractions of memory displayed on the signs of nature only on casual and fragmentary, such as line, square and circle: those initials and abstract forms located in abstraction give rise to the first signs of primordial architecture, so that squares and circles, lifting two prisms and spheroids or spherical surfaces appear in tombs and elementary buildings the first few times, and that primordial geometry overlaps the landscape in which it is inserted. It is also true that these forms are approximate objects found in the physical environment, limb lines, branches and limbs; straight in some crystals angles observed by cutting or circular in stars and craters forms, but only the approximate shape and not its abstraction, just as the hieroglyphic of a falcon to represent the idea of surveillance presents the good view, or even the image of the eye itself replace the idea of the all-seeing God. The election was resolved, after many attempts and approaches, with the adoption of the right angle as an easy trick for stake through the sacred triangle 3:4:5 already used to build the pyramids of Egypt, and also gave rise to most of the old urbanism tend coexist with the primary circular form type called "Tholo". That frame homer was one of the fill patterns of common and compact space, and that was probably the reason why in much later times was adopted as a permanent effective form of organization against the topological disorder that seeks the set of association circular plants. Another issue is that these paradigmatic and unknown primary concepts - that converge at the origin of civilizations - then become something canon, through use. The canon itself is something ideal, as a rule for objects of a reality that has been created only as an indication of the ca-non, something abstract that has strict proportions are always the same and not due to rational criteria: be absurd however seek the Greek canon among the temples of time as some did, because the buildings only approximate the canonical examples and for that reason we speak of "doric from Parthenon" which is different from Aegina or Paestum, being all clearly doric. However, this idea is useful to scholar, the architectural theorist and historian, although it reflects only a vague idea of what happens beyond the book. Another issue is the subtlety of the examples of architecture, just as the bodies of living beings are never symmetrical, even if they involve a general symmetrical design derived from the conditions of cell division, the supposedly canonical buildings where specialists were inspired to define the classical orders do not have that exact modular symmetry, but a general model adapted to the ever changing location of the site shaping the landscape in which they are inserted. The light of the various bays of the Parthenon is slightly different, but keep a clear sense of axial symmetry, so that those "orders" that formed the theory of architecture are just a beautiful sectoral interpretation of ancient architecture developed by writers of the Renaissance and later by neoclassical. It seems, however, that craving for clarity of language canon as is inherent to the development of architecture, a lingua franca that tends to avoid scattering among mortals by the ancient gods, so if it was not possible the heaven of the gods sought the language of the Earth was at least intelligible to be understood. Power structure has always required a certain level of organization, and the instructions are clearly understood: it seems that in our times these ancient canons were replaced by obedience to abstract rules, issued by institutions also somewhat abstract and have funny names made up acronym, although not well known virtue. The current canon consists of "rules" that leave some quiet, although it seems that maybe all that interweaves-do formally serving as anaesthesia for the social body also seeks the destruction of forests in forms of huge mountains of bureaucratic paper. For many centuries were other rules, in the form of a canon which no one could escape: still using royalties or without them, civilization flourished in architecture from the earliest cave shelters or shelters and the primordial reality was setting mode in landscape. As noted above, this is a return trip in which both confused and subsumed: the management of forms and languages of architecture over time would allow the distinction between the field where chaos reigns and city, where order reigns theoretically creating a divorce that lasted millennia and still persists. This opposition also generate a form of landscape and a series of symbolic and sacred uses of architectural elements - such as doors and walls - which have remained conceptually although today the cities no longer having physical walls or doors close during the night. In this sense, the architecture could define the landscape, architecture is not only understood as the buildings themselves, but the fact of the manifestation of the man on the premises, so you can not speak without his landscape architecture, the same so we can not speak of landscape without men. For this reason, the thesis discusses architecture and landscape, but more particularly to the fact of architecture in defining landscape, as the facts of architectural or not will determine the quality and qualification of a landscape. One should begin first a definition of what is understood and has been commonly understood by landscape: equally, and given the context in which it places the own thesis work, the issue only be limited to what is understood as western culture, for the development of civilization always has a local colour that makes the analysis of a phenomenon of this magnitude have different connotations in different areas of civilization. Similarly, and given also that the landscape is constructed through all cultural manifestations, will sometimes necessary to investigate other non-architectural disciplines to understand the scope of some cases in which the architectural remains have disappeared, or the remaining few traces. Such a broad definition of architecture take to write a story about all of Western culture and for this reason only been sketched ideas about the appearance of these signs on the landscape based on ancient and primitive elements are repeated insistently and leading the varied repertoire of Western architecture shaping ideas about how the media and the world is perceived and inhabited.

Loss of ovarian function promotes angiogenesis in human ovarian carcinoma

We show here that elevated levels of gonadotropins (luteinizing hormone and follicle stimulating hormone), as found in menopause or after ovariectomy, promote growth of human ovarian carcinoma by induction of tumor angiogenesis. Human epithelial ovarian cancer tumors progressed faster in ovariectomized mice. This induced growth could be attributed to the elevated levels of gonadotropins associated with loss of ovarian function because direct administration of gonadotropins also was effective in promoting tumor progression in vivo. On the other hand, gonadotropins had no direct effect on the proliferation of human ovarian cancer cells in vitro. Using MRI, we demonstrated that ovariectomy significantly (P < 0.02) induces neovascularization of human ovarian carcinoma spheroids implanted in nude mice. Moreover, conditioned medium of gonadotropin-treated human ovarian carcinoma cells showed increased mitogenic activity to bovine endothelial cells, and this activity could be blocked by neutralizing antibodies against luteinizing hormone and against vascular endothelial growth factor. Accordingly, gonadotropin stimulation resulted in a dose-dependent-induced expression of vascular endothelial growth factor in monolayer culture as well as in the outer proliferating cells of human ovarian cancer spheroids. These results demonstrate the significance of the elevated levels of gonadotropins, as found in menopause and in all ovarian cancer patients, on the progression of ovarian cancer and could explain the protective effect of estrogen replacement therapy. Based on these results, we suggest that hormonal therapy aimed at lowering the circulating levels of gonadotropins may possibly prolong remission in ovarian cancer by extending tumor dormancy.

Transgenic mice carrying a human mutant superoxide dismutase transgene develop neuronal cytoskeletal pathology resembling human amyotrophic lateral sclerosis lesions.

Mutations in the human Cu,Zn superoxide dismutase gene (SOD1) are found in 20% of kindreds with familial amyotrophic lateral sclerosis. Transgenic mice (line G1H) expressing a human SOD1 containing a mutation of Gly-93 --> Ala (G93A) develop a motor neuron disease similar to familial amyotrophic lateral sclerosis, but transgenic mice (line N1029) expressing a wild-type human SOD1 transgene do not. Because neurofilament (NF)-rich inclusions in spinal motor neurons are characteristic of amyotrophic lateral sclerosis, we asked whether mutant G1H and/or N1029 mice develop similar NF lesions. NF inclusions (i.e., spheroids, Lewy body-like inclusions) were first detected in spinal cord motor neurons of the G1H mice at 82 days of age about the time these mice first showed clinical evidence of disease. Other neuronal intermediate filament proteins (alpha-internexin, peripherin) also accumulated in these spheroids. The onset of accumulations of ubiquitin immunoreactivity in the G1H mice paralleled the emergence of vacuoles and NF-rich spheroids in neurons, but they did not colocalize exclusively with spheroids. In contrast, NF inclusions were not seen in the N1029 mice until they were 132 days old, and ubiquitin immunoreactivity was not increased in the N1029 mice even at 199 days of age. Astrocytosis in spinal cord was associated with a marked increase in glial fibrillary acidic protein immunoreactivity in the G1H mice, but not in the N1029 mice. Finally, comparative studies revealed a striking similarity between the cytoskeletal pathology in the G1H transgenic mice and in patients with amyotrophic lateral sclerosis. These findings link a specific SOD1 mutation with alterations in the neuronal cytoskeleton of patients with amyotrophic lateral sclerosis. Thus, neuronal cytoskeletal abnormalities may be implicated in the pathogenesis of human familial amyotrophic lateral sclerosis.

The star formation history of CALIFA galaxies: Radial structures

We have studied the radial structure of the stellar mass surface density (μ∗) and stellar population age as a function of the total stellar mass and morphology for a sample of 107 galaxies from the CALIFA survey. We applied the fossil record method based on spectral synthesis techniques to recover the star formation history (SFH), resolved in space and time, in spheroidal and disk dominated galaxies with masses from 10^9 to 10^12 M_⊙. We derived the half-mass radius, and we found that galaxies are on average 15% more compact in mass than in light. The ratio of half-mass radius to half-light radius (HLR) shows a dual dependence with galaxy stellar mass; it decreases with increasing mass for disk galaxies, but is almost constant in spheroidal galaxies. In terms of integrated versus spatially resolved properties, we find that the galaxy-averaged stellar population age, stellar extinction, and μ_∗ are well represented by their values at 1 HLR. Negative radial gradients of the stellar population ages are present in most of the galaxies, supporting an inside-out formation. The larger inner (≤1 HLR) age gradients occur in the most massive (10^11 M_⊙) disk galaxies that have the most prominent bulges; shallower age gradients are obtained in spheroids of similar mass. Disk and spheroidal galaxies show negative μ∗ gradients that steepen with stellar mass. In spheroidal galaxies, μ∗ saturates at a critical value (~7 × 10^2 M_⊙/pc^2 at 1 HLR) that is independent of the galaxy mass. Thus, all the massive spheroidal galaxies have similar local μ_∗ at the same distance (in HLR units) from the nucleus. The SFH of the regions beyond 1 HLR are well correlated with their local μ_∗, and follow the same relation as the galaxy-averaged age and μ_∗; this suggests that local stellar mass surface density preserves the SFH of disks. The SFH of bulges are, however, more fundamentally related to the total stellar mass, since the radial structure of the stellar age changes with galaxy mass even though all the spheroid dominated galaxies have similar radial structure in μ_∗. Thus, galaxy mass is a more fundamental property in spheroidal systems, while the local stellar mass surface density is more important in disks.

The tidal and other problems,

At head of title: Contributions to cosmogony and the fundamental problems of geology.