Dendrimers as potential platform in nanotechnology-based drug delivery systems

The dendrimers of poly (amidoamine) (PAMAM) are nanoparticles which have proven succeed in transporting drugs due to high solubility, low toxicity and ability to control drugs release. Studies have explored the biological potential of dendrimers such as to transport genes, development of vaccines, antiviral, antibacterial and anticancer therapies. This review of literature on the PAMAM dendrimers discusses the architecture and general construction of dendrimers and intrinsic properties of the PAMAM. This study also describes how the PAMAM interact with many drugs and the potential of these macromolecules as well as drug nanocarriers in transdermal routes of administration, ocular, respiratory, oral and intravenous administration. Dendrimers promises good future prospects for the biomedicine.

Inward-Inclined Implant Platform for the Amplified Platform-Switching Concept: 18-Month Follow-up Report of a Prospective Randomized Matched-Pair Controlled Trial

Purpose: This prospective randomized matched-pair controlled trial aimed to evaluate marginal bone levels and soft tissue alterations at implants restored according to the platform-switching concept with a new inward-inclined platform and compare them with external-hexagon implants. Materials and Methods: Traditional external-hexagon (control group) implants and inward-inclined platform implants (test group), all with the same implant body geometry and 13 mm in length, were inserted in a standardized manner in the posterior maxillae of 40 patients. Radiographic bone levels were measured by two independent examiners after 6, 12, and 18 months of prosthetic loading. Buccal soft tissue height was measured at the time of abutment connection and 18 months later. Results: After 18 months of loading, all 80 implants were clinically osseointegrated in the 40 participating patients. Radiographic evaluation showed mean bone losses of 0.5 +/- 0.1 mm (range, 0.3 to 0.7 mm) and 1.6 +/- 0.3 mm (range, 1.1 to 2.2 mm) for test and control implants, respectively. Soft tissue height showed a significant mean decrease of 2.4 mm in the control group, compared to 0.6 mm around the test implants. Conclusions: After 18 months, significantly greater bone loss was observed at implants restored according to the conventional external-hexagon protocol compared to the platform-switching concept. In addition, decreased soft tissue height was associated with the external-hexagon implants versus the platform-switched implants. INT J ORAL MAXILLOFAC IMPLANTS 2012;27:927-934.

Climate impact studies of sediment transport on the Adriatic coastal zone

The study of the impact of climate change on the environment has been based, until very recently, on an global approach, whose interest from a local point of view is very limited. This thesis, on the contrary, has treated the study of the impact of climate change in the Adriatic Sea basin following a twofold strategy of regionalization and integration of numerical models in order to reproduce the present and future scenarios of the system through a more and more realistic and solid approach. In particular the focus of the study was on the impact on the physical environment and on the sediment transport in the basin. This latter is a very new and original issue, to our knowledge still uninvestigated. The study case of the coastal area of Montenegro was particularly studied, since it is characterized by an important supply of sediment through the Buna/Bojana river, second most important in the Adriatic basin in terms of flow. To do this, a methodology to introduce the tidal processes in a baroclinic primitive equations Ocean General Circulation Model was applied and tidal processes were successfully reproduced in the Adriatic Sea, analyzing also the impacts they have on the mean general circulation, on salt and heat transport and on mixing and stratification of the water column in the different seasons of the year. The new hydrodynamical model has been further coupled with a wave model and with a river and sea sediment transport model, showing good results in the reproduction of sediment transport processes. Finally this complex coupled platform was integrated in the period 2001-2030 under the A1B scenario of IPCC, and the impact of climate change on the physical system and on sediment transport was preliminarily evaluated.

Systematic studies of protein immobilization by surface plasmon field-enhanced fluorescence spectroscopy

The research interest of this study is to investigate surface immobilization strategies for proteins and other biomolecules by the surface plasmon field-enhanced fluorescence spectroscopy (SPFS) technique. The recrystallization features of the S-layer proteins and the possibility of combining the S-layer lattice arrays with other functional molecules make this protein a prime candidate for supramolecular architectures. The recrystallization behavior on gold or on the secondary cell wall polymer (SCWP) was recorded by SPR. The optical thicknesses and surface densities for different protein layers were calculated. In DNA hybridization tests performed in order to discriminate different mismatches, recombinant S-layer-streptavidin fusion protein matrices showed their potential for new microarrays. Moreover, SCWPs coated gold chips, covered with a controlled and oriented assembly of S-layer fusion proteins, represent an even more sensitive fluorescence testing platform. Additionally, S-layer fusion proteins as the matrix for LHCII immobilization strongly demonstrate superiority over routine approaches, proving the possibility of utilizing them as a new strategy for biomolecular coupling. In the study of the SPFS hCG immunoassay, the biophysical and immunological characteristics of this glycoprotein hormone were presented first. After the investigation of the effect of the biotin thiol dilution on the coupling efficiently, the interfacial binding model including the appropriate binary SAM structure and the versatile streptavidin-biotin interaction was chosen as the basic supramolecular architecture for the fabrication of a SPFS-based immunoassay. Next, the affinity characteristics between different antibodies and hCG were measured via an equilibrium binding analysis, which is the first example for the titration of such a high affinity interaction by SPFS. The results agree very well with the constants derived from the literature. Finally, a sandwich assay and a competitive assay were selected as templates for SPFS-based hCG detection, and an excellent LOD of 0.15 mIU/ml was attained via the “one step” sandwich method. Such high sensitivity not only fulfills clinical requirements, but is also better than most other biosensors. Fully understanding how LHCII complexes transfer the sunlight energy directionally and efficiently to the reaction center is potentially useful for constructing biomimetic devices as solar cells. After the introduction of the structural and the spectroscopic features of LHCII, different surface immobilization strategies of LHCII were summarized next. Among them the strategy based on the His-tag and the immobilized metal (ion) affinity chromatography (IMAC) technique were of great interest and resulted in different kinds of home-fabricated His-tag chelating chips. Their substantial protein coupling capacity, maintenance of high biological activity and a remarkably repeatable binding ability on the same chip after regeneration was demonstrated. Moreover, different parameters related to the stability of surface coupled reconstituted complexes, including sucrose, detergent, lipid, oligomerization, temperature and circulation rate, were evaluated in order to standardize the most effective immobilization conditions. In addition, partial lipid bilayers obtained from LHCII contained proteo-liposomes fusion on the surface were observed by the QCM technique. Finally, the inter-complex energy transfer between neighboring LHCIIs on a gold protected silver surface by excitation with a blue laser (λ = 473nm) was recorded for the first time, and the factors influencing the energy transfer efficiency were evaluated.

Systematic studies of the interaction between amyloid beta-protein and lipids

The amyloid peptide (Aß), a normal constituent of neuronal and non-neuronal cells, has been shown to be a major component of the extracellular plaque of Alzheimer’s disease (AD). The interaction of Aß peptides with the lipid matrix of neuronal cell membranes plays an important role in the pathogenesis of AD. In this study, we have developed peptide-tethered artificial lipid membranes by the Langmuir-Blodgett and Langmuir-Schaefer methods. Anti-Aß40-mAb labeled with a fluorophore was used to probe the Aß40 binding to the model membrane system. Systematic studies on the antibody or Aß-membrane interactions were carried out in our model systems by Surface Plasmon Field-Enhanced Fluorescence Spectroscopy (SPFS). Aß adsorption is critically determined by the lipid composition of the membranes. Aß specifically binds with membranes of sphingomyelin, and this preferential adsorption was markedly amplified by the addition of sterols (cholesterol or 25-OH-Chol). Fluorescence microscopy indicated that 25-OH-Chol could also form micro-domains with sphingomyelin as cholesterol does at the conditions used for the built-up of the model membranes. Our findings suggest that micro-domains composed of sphingomyelin and the sterols could be the binding sites of Aß and the role of sphingomyelin in AD should receive much more attention. The artificial membranes provide a novel platform for the study on AD, and SPFS is a potential tool for detecting Aß-membrane interaction. Numerous investigations indicate that the ability of Aß to form fibrils is considerably dependent upon the levels of ß-sheet structure adopted by Aß. Membrane-mediated conformational transition of Aß has been demonstrated. In this study, we focus on the interaction of Aß and the membranes composed of POPC/SM/25-OH-Chol (2:1:1). The artificial membrane system was established by the methods as described above. Immunoassy based on a pair of monoclonal antibodies (mAbs) against different epitopes was employed to detect the orientation of the Aß at the model membranes. Kinetics of antibody-Aß binding was determined by surface plasmon field-enhanced fluorescence spectroscopy (SPFS). The attempt has also been made to probe the change in the conformation of Aß using SPFS combined with immunoassay. Melatonin was employed to induce the conformational change of Aß. The orientation and the conformational change of Aß are evaluated by analysing kinetic/affinity parameters. This work provides novel insight into the investigation on the structure of Aß at the membrane surface.

Surface plasmon fluorescence spectroscopy and microscopy studies for biomolecular interaction studies

ABSTARCT Biotechnology has enabled the modification of agricultural materials in a very precise way. Crops have been modified through the insertion of new traits or the inhibition of existing gene functions, named Genetically Modified Organism (GMO), and resulted in improved tolerance of herbicide and/or increased resistance against pests, viruses and fungi. Commercial cultivation of GMO started in 1996 and increased rapidly in 2003 according to a recently released report by the International Service for the Acquisition of Agri-Biotech Applications (ISAAA), depicted continuing consumer resistance in Europe and other part of the world. Upon these developments, the European Union regulations mandated labeling of GMOs containing food and as a consequence, the labeling of GMO containing product in the case of exceeding the1% threshold of alien DNA is required. The aim of the study is to be able to detect and quantify the GMO from the mixture of natural food components. The surface plasmon resonance (SPR) technique combined with fluorescence was used for this purpose. During the presented studies, two key issues are addressed and tried to solve; what is the best strategy to design and built an interfacial architecture of a probe oligonucletide layer either on a two dimensional surface or on an array platform; and what is the best detection method allowing for a sensitive monitoring of the hybridisation events. The study includes two parts: first part includes characterization of different PNAs on a 2D planar surface by defining affinity constants using the very well established optical method “Surface Plasmon Fluorescence Spectroscopy”(SPFS) and on the array platform by “Surface Plasmon Fluorescence Microscopy” (SPFM), and at the end comparison of the sensitivity of these two techniques. The second part is composed of detection of alien DNA in food components by using DNA and PNA catcher probes on the array platform in real-time by SPFM.

Nucleation studies on graphics processing units

Ein System in einem metastabilen Zustand muss eine bestimmte Barriere in derrnfreien Energie überwinden um einen Tropfen der stabilen Phase zu formen.rnHerkömmliche Untersuchungen nehmen hierbei kugelförmige Tropfen an. Inrnanisotropen Systemen (wie z.B. Kristallen) ist diese Annahme aber nicht ange-rnbracht. Bei tiefen Temperaturen wirkt sich die Anisotropie des Systems starkrnauf die freie Energie ihrer Oberfläche aus. Diese Wirkung wird oberhalb derrnAufrauungstemperatur T R schwächer. Das Ising-Modell ist ein einfaches Mo-rndell, welches eine solche Anisotropie aufweist. Wir führen großangelegte Sim-rnulationen durch, um die Effekte, die mit einer endlichen Simulationsbox ein-rnhergehen, sowie statistische Ungenauigkeiten möglichst klein zu halten. DasrnAusmaß der Simulationen die benötigt werden um sinnvolle Ergebnisse zu pro-rnduzieren, erfordert die Entwicklung eines skalierbaren Simulationsprogrammsrnfür das Ising-Modell, welcher auf verschiedenen parallelen Architekturen (z.B.rnGrafikkarten) verwendet werden kann. Plattformunabhängigkeit wird durch ab-rnstrakte Schnittstellen erreicht, welche plattformspezifische Implementierungs-rndetails verstecken. Wir benutzen eine Systemgeometrie die es erlaubt eine Ober-rnfläche mit einem variablen Winkel zur Kristallebene zu untersuchen. Die Ober-rnfläche ist in Kontakt mit einer harten Wand, wobei der Kontaktwinkel Θ durchrnein Oberflächenfeld eingestellt werden kann. Wir leiten eine Differenzialglei-rnchung ab, welche das Verhalten der freien Energie der Oberfläche in einemrnanisotropen System beschreibt. Kombiniert mit thermodynamischer Integrationrnkann die Gleichung benutzt werden, um die anisotrope Oberflächenspannungrnüber einen großen Winkelbereich zu integrieren. Vergleiche mit früheren Mes-rnsungen in anderen Geometrien und anderen Methoden zeigen hohe Überein-rnstimung und Genauigkeit, welche vor allem durch die im Vergleich zu früherenrnMessungen wesentlich größeren Simulationsdomänen erreicht wird. Die Temper-rnaturabhängigkeit der Oberflächensteifheit κ wird oberhalb von T R durch diernKrümmung der freien Energie der Oberfläche für kleine Winkel gemessen. DiesernMessung lässt sich mit Simulationsergebnissen in der Literatur vergleichen undrnhat bessere Übereinstimmung mit theoretischen Voraussagen über das Skalen-rnverhalten von κ. Darüber hinaus entwickeln wir ein Tieftemperatur-Modell fürrndas Verhalten um Θ = 90 Grad weit unterhalb von T R. Der Winkel bleibt bis zu einemrnkritischen Feld H C quasi null; oberhalb des kritischen Feldes steigt der Winkelrnrapide an. H C wird mit der freien Energie einer Stufe in Verbindung gebracht,rnwas es ermöglicht, das kritische Verhalten dieser Größe zu analysieren. Die harternWand muss in die Analyse einbezogen werden. Durch den Vergleich freier En-rnergien bei geschickt gewählten Systemgrößen ist es möglich, den Beitrag derrnKontaktlinie zur freien Energie in Abhängigkeit von Θ zu messen. Diese Anal-rnyse wird bei verschiedenen Temperaturen durchgeführt. Im letzten Kapitel wirdrneine 2D Fluiddynamik Simulation für Grafikkarten parallelisiert, welche u. a.rnbenutzt werden kann um die Dynamik der Atmosphäre zu simulieren. Wir im-rnplementieren einen parallelen Evolution Galerkin Operator und erreichen

Influence of platform switching on bone-level alterations: a three-year randomized clinical trial

The concept of platform switching has been introduced to implant dentistry based on clinical observations of reduced peri-implant crestal bone loss. However, published data are controversial, and most studies are limited to 12 months. The aim of the present randomized clinical trial was to test the hypothesis that platform switching has a positive impact on crestal bone-level changes after 3 years. Two implants with a diameter of 4 mm were inserted crestally in the posterior mandible of 25 patients. The intraindividual allocation of platform switching (3.3-mm platform) and the standard implant (4-mm platform) was randomized. After 3 months of submerged healing, single-tooth crowns were cemented. Patients were followed up at short intervals for monitoring of healing and oral hygiene. Statistical analysis for the influence of time and platform type on bone levels employed the Brunner-Langer model. At 3 years, the mean radiographic peri-implant bone loss was 0.69 ± 0.43 mm (platform switching) and 0.74 ± 0.57 mm (standard platform). The mean intraindividual difference was 0.05 ± 0.58 mm (95% confidence interval: -0.19, 0.29). Crestal bone-level alteration depended on time (p < .001) but not on platform type (p = .363). The present randomized clinical trial could not confirm the hypothesis of a reduced peri-implant crestal bone loss, when implants had been restored according to the concept of platform switching.

Next-generation tissue microarray (ngTMA) increases the quality of biomarker studies: an example using CD3, CD8, and CD45RO in the tumor microenvironment of six different solid tumor types

Background Tissue microarray (TMA) technology revolutionized the investigation of potential biomarkers from paraffin-embedded tissues. However, conventional TMA construction is laborious, time-consuming and imprecise. Next-generation tissue microarrays (ngTMA) combine histological expertise with digital pathology and automated tissue microarraying. The aim of this study was to test the feasibility of ngTMA for the investigation of biomarkers within the tumor microenvironment (tumor center and invasion front) of six tumor types, using CD3, CD8 and CD45RO as an example. Methods Ten cases each of malignant melanoma, lung, breast, gastric, prostate and colorectal cancers were reviewed. The most representative H&E slide was scanned and uploaded onto a digital slide management platform. Slides were viewed and seven TMA annotations of 1 mm in diameter were placed directly onto the digital slide. Different colors were used to identify the exact regions in normal tissue (n = 1), tumor center (n = 2), tumor front (n = 2), and tumor microenvironment at invasion front (n = 2) for subsequent punching. Donor blocks were loaded into an automated tissue microarrayer. Images of the donor block were superimposed with annotated digital slides. Exact annotated regions were punched out of each donor block and transferred into a TMA block. 420 tissue cores created two ngTMA blocks. H&E staining and immunohistochemistry for CD3, CD8 and CD45RO were performed. Results All 60 slides were scanned automatically (total time < 10 hours), uploaded and viewed. Annotation time was 1 hour. The 60 donor blocks were loaded into the tissue microarrayer, simultaneously. Alignment of donor block images and digital slides was possible in less than 2 minutes/case. Automated punching of tissue cores and transfer took 12 seconds/core. Total ngTMA construction time was 1.4 hours. Stains for H&E and CD3, CD8 and CD45RO highlighted the precision with which ngTMA could capture regions of tumor-stroma interaction of each cancer and the T-lymphocytic immune reaction within the tumor microenvironment. Conclusion Based on a manual selection criteria, ngTMA is able to precisely capture histological zones or cell types of interest in a precise and accurate way, aiding the pathological study of the tumor microenvironment. This approach would be advantageous for visualizing proteins, DNA, mRNA and microRNAs in specific cell types using in situ hybridization techniques.

A novel comparative research platform designed to determine the functional significance of tree species diversity in European forests

One of the current advances in functional biodiversity research is the move away from short-lived test systems towards the exploration of diversity-ecosystem functioning relationships in structurally more complex ecosystems. In forests, assumptions about the functional significance of tree species diversity have only recently produced a new generation of research on ecosystem processes and services. Novel experimental designs have now replaced traditional forestry trials, but these comparatively young experimental plots suffer from specific difficulties that are mainly related to the tree size and longevity. Tree species diversity experiments therefore need to be complemented with comparative observational studies in existing forests. Here we present the design and implementation of a new network of forest plots along tree species diversity gradients in six major European forest types: the FunDivEUROPE Exploratory Platform. Based on a review of the deficiencies of existing observational approaches and of unresolved research questions and hypotheses, we discuss the fundamental criteria that shaped the design of our platform. Key features include the extent of the species diversity gradient with mixtures up to five species, strict avoidance of a dilution gradient, special attention to community evenness and minimal covariation with other environmental factors. The new European research platform permits the most comprehensive assessment of tree species diversity effects on forest ecosystem functioning to date since it offers a common set of research plots to groups of researchers from very different disciplines and uses the same methodological approach in contrasting forest types along an extensive environmental gradient. (C) 2013 Elsevier GmbH. All rights reserved.

Concept, design and implementation of a cardiovascular gene-centric 50 k SNP array for large-scale genomic association studies.

A wealth of genetic associations for cardiovascular and metabolic phenotypes in humans has been accumulating over the last decade, in particular a large number of loci derived from recent genome wide association studies (GWAS). True complex disease-associated loci often exert modest effects, so their delineation currently requires integration of diverse phenotypic data from large studies to ensure robust meta-analyses. We have designed a gene-centric 50 K single nucleotide polymorphism (SNP) array to assess potentially relevant loci across a range of cardiovascular, metabolic and inflammatory syndromes. The array utilizes a "cosmopolitan" tagging approach to capture the genetic diversity across approximately 2,000 loci in populations represented in the HapMap and SeattleSNPs projects. The array content is informed by GWAS of vascular and inflammatory disease, expression quantitative trait loci implicated in atherosclerosis, pathway based approaches and comprehensive literature searching. The custom flexibility of the array platform facilitated interrogation of loci at differing stringencies, according to a gene prioritization strategy that allows saturation of high priority loci with a greater density of markers than the existing GWAS tools, particularly in African HapMap samples. We also demonstrate that the IBC array can be used to complement GWAS, increasing coverage in high priority CVD-related loci across all major HapMap populations. DNA from over 200,000 extensively phenotyped individuals will be genotyped with this array with a significant portion of the generated data being released into the academic domain facilitating in silico replication attempts, analyses of rare variants and cross-cohort meta-analyses in diverse populations. These datasets will also facilitate more robust secondary analyses, such as explorations with alternative genetic models, epistasis and gene-environment interactions.

Structure/affinity studies in the bicyclo-DNA series: Synthesis and properties of oligonucleotides containing bcen-T and iso-tricyclo-T nucleosides

We present the synthesis of the two novel nucleosides iso-tc-T and bcen-T, belonging to the bicyclo-/tricyclo-DNA molecular platform. In both modifications the torsion around C6’–C7’ within the carbocyclic ring is planarized by either the presence of a C6’–C7’ double bond or a cyclopropane ring. Structural analysis of these two nucleosides by X-ray analysis reveals a clear preference of torsion angle γ for the gauche orientation with the furanose ring in a near perfect 2’-endo conformation. Both modifications were incorporated into oligodeoxynucleotides and their thermal melting behavior with DNA and RNA as complements was assessed. We found that the iso-tc-T modification was significantly more destabilizing in duplex formation compared to the bcen-T modification. In addition, duplexes with complementary RNA were less stable as compared to duplexes with DNA as complement. A structure/affinity analysis, including the already known bc-T and tc-T modifications, does not lead to a clear correlation of the orientation of torsion angle γ with DNA or RNA affinity. There is, however, some correlation between furanose conformation (N- or S-type) and affinity in the sense that a preference for a 3’-endo like conformation is associated with a preference for RNA as complement. As a general rule it appears that Tm data of single modifications with nucleosides of the bicyclo-/tricyclo-DNA platform within deoxyoligonucleotides are not predictive for the stability of fully modified oligonucleotides.

A next-generation tissue microarray (ngTMA) protocol for biomarker studies

Biomarker research relies on tissue microarrays (TMA). TMAs are produced by repeated transfer of small tissue cores from a 'donor' block into a 'recipient' block and then used for a variety of biomarker applications. The construction of conventional TMAs is labor intensive, imprecise, and time-consuming. Here, a protocol using next-generation Tissue Microarrays (ngTMA) is outlined. ngTMA is based on TMA planning and design, digital pathology, and automated tissue microarraying. The protocol is illustrated using an example of 134 metastatic colorectal cancer patients. Histological, statistical and logistical aspects are considered, such as the tissue type, specific histological regions, and cell types for inclusion in the TMA, the number of tissue spots, sample size, statistical analysis, and number of TMA copies. Histological slides for each patient are scanned and uploaded onto a web-based digital platform. There, they are viewed and annotated (marked) using a 0.6-2.0 mm diameter tool, multiple times using various colors to distinguish tissue areas. Donor blocks and 12 'recipient' blocks are loaded into the instrument. Digital slides are retrieved and matched to donor block images. Repeated arraying of annotated regions is automatically performed resulting in an ngTMA. In this example, six ngTMAs are planned containing six different tissue types/histological zones. Two copies of the ngTMAs are desired. Three to four slides for each patient are scanned; 3 scan runs are necessary and performed overnight. All slides are annotated; different colors are used to represent the different tissues/zones, namely tumor center, invasion front, tumor/stroma, lymph node metastases, liver metastases, and normal tissue. 17 annotations/case are made; time for annotation is 2-3 min/case. 12 ngTMAs are produced containing 4,556 spots. Arraying time is 15-20 hr. Due to its precision, flexibility and speed, ngTMA is a powerful tool to further improve the quality of TMAs used in clinical and translational research.

p-medicine: a medical informatics platform for integrated large scale heterogeneous patient data

Secure access to patient data is becoming of increasing importance, as medical informatics grows in significance, to both assist with population health studies, and patient specific medicine in support of treatment. However, assembling the many different types of data emanating from the clinic is in itself a difficulty, and doing so across national borders compounds the problem. In this paper we present our solution: an easy to use distributed informatics platform embedding a state of the art data warehouse incorporating a secure pseudonymisation system protecting access to personal healthcare data. Using this system, a whole range of patient derived data, from genomics to imaging to clinical records, can be assembled and linked, and then connected with analytics tools that help us to understand the data. Research performed in this environment will have immediate clinical impact for personalised patient healthcare.

Decodificar Sejima‐SANAA: Estrategias formales y estructurales en su trayectoria de proyecto, 1987 a 2010; de la Casa Platform I al Centro Rolex

La Tesis decodifica una selección de veinte proyectos representativos de Sejima-SANAA, desde su primer proyecto construido, la Casa Platform I, 1987, hasta el Centro Rolex, 2010, año en que Sejima y Nishizawa –SANAA- reciben el Premio Pritzker. De los veinte proyectos once son de Sejima: Casa Platform I, Casa Platform II, Residencia de Mujeres, Casa N, Pachinco Parlor I, Villa en el Bosque, Comisaría en Chofu, Casa Y, Apartamentos en Gifu, Edificio de equipamientos en la Expo Tokio 96, Pachinko Parlor III; y nueve de SANAA: edificio Multimedia en Oogaki, estudio de viviendas metropolitanas,Park Café en Koga, De Kunstlinie en Almere, Museo de Kanazawa, Pabellón de Toledo, Escuela de Zollverein, Casa Flor y Centro Rolex. La decodificación lee la obra de Sejima-SANAA a la inversa para ‘reconstruir’, en un ejercicio de simulación ficticia, una versión verosímil y coherente de los que podrían haber sido sus procesos proyectuales; podrían, porque los verdaderos son imposibles de dilucidar. Los que se proponen se pretenden exclusivamente verosímiles y plausibles. Con ello se pretende contribuir al entendimiento y comprensión de la arquitectura de Sejima-SANAA y, tangencialmente y en menor medida, a la teoría sobre el ejercicio proyectual arquitectónico. La decodificación se centra en dos aspectos concretos: la forma arquitectónica y el papel proyectual de la estructura portante. Ambas decodificaciones se extienden inevitablemente a otros aspectos relacionados, como, por ejemplo, la naturaleza del espacio arquitectónico. El procedimiento de investigación partió de una descripción objetiva y pormenorizada de los significantes formales y estructurales de cada proyecto desde su propia configuración física y geométrica. Esa descripción ‘objetiva’, llevada al límite, permitió que afloraran estructuras conceptuales y lógicas subyacentes de cada proyecto. Unida a interpretación crítica, –mediante su relación y confrontación con otras arquitecturas y otros modos de hacer conocidos- permitió trazar la reconstitución ficticia que persigue la decodificación. Ese trabajo se materializó en veinte ensayos críticos y se acompañó de un conjunto de otros textos sobre temas sugeridos o reclamados por el proceso de investigación. El conjunto de todos esos textos constituye el material de trabajo de la tesis. A partir de ahí, con una visión de conjunto, la tesis identifica una trayectoria de estrategias formales y una trayectoria de estrategias proyectuales relacionadas con lo portante. Juntas conforman el grueso de la tesis que se expone en los cuatro capítulos centrales. Los precede un capítulo introductorio que expone el recorrido biográfico de K. Sejima y la trayectoria profesional de Sejima-SANAA; y los siguen de unos textos transversales sobre forma, lugar y espacio. La tesis termina con una síntesis de sus conclusiones. Las estrategias formales se exponen en tres capítulos. El primero, ‘Primeras estrategias formales’ agrupa proyectos de la primera etapa de Sejima. El segundo capítulo está dedicado enteramente al proyecto de los apartamentos en Gifu, 1994-98, que según esta tesis, supuso un importante punto de inflexión en la trayectoria de Sejima; tanto el tercer capítulo lleva por nombre ‘Estrategias formales después de Gifu’ y recoge los proyectos que le siguieron. Las ‘Primeras estrategias formales’, varias y balbucientes, se mueven en general en torno a dos modos o procedimientos de composición, bien conocidos: por partes y sistemático. Éste última inicia en la trayectoria de SANAA un aspecto que va a ser relevante de aquí en adelante: entender el proyecto como propuesta genérica en la que, más allá de su realidad específica y tangible, subyace una lógica, en cada proyecto la suya, extrapolable a otros lugares, otras dimensiones, incluso otros programas: cada proyecto podría dar lugar a otros proyectos de la misma familia. La composición sistemática incluye, entre otros, la Casa Platform II, basada en la definición de un elemento constructivo, y la formulación de unas leyes de repetición y de posibles modos de agrupación. Incluye también la Residencia de Mujeres Saishunkan Seiyaku- proyecto que lanzó a Sejima a la fama internacional-, que también sería un sistema, pero distinto: basado en la repetición regular de una serie de elementos a lo largo de una directriz generando un hipotético contenedor infinito del que el proyecto sería tan solo un fragmento. La estrategia formal del edificio de Gifu ahondaría en la voluntad genérica del proyecto, adoptando la lógica de un juego. El proyecto sería una partida del juego, pero no la única posible, podrían jugarse otras. Esta hipótesis del juego está verificada en ‘El Juego de Gifu’ que - tras formular el juego identificando sus elementos (tablero y fichas), reglas y procedimientos- juega una partida: la que habría dado lugar al edificio proyectado por Sejima. Gifu extiende el concepto de ‘repetir’ un elemento constructivo a la de repetir un patrón espacial, lo que conlleva: la desvinculación entre forma y función; y un nuevo concepto de flexibilidad, que deja de referirse al uso flexible del edificio construido para pertenecer al momento proyectual en que se asignan funciones específicas a los patrones espaciales. Esta tesis propone que esa asignación de funciones sería uno de los últimos eslabones del proceso proyectual, algo opuesto a la premisa moderna de “la forma sigue a la función”. Las estrategias formales ‘Después de Gifu’ tienen también lógicas de juego, pero cada estrategia responde a un juego distinto, como dejan entrever sus nombres: ‘Tableros de Juego’, que con distintos grados de madurez estaría presente en varios proyectos; ‘Elementos de Catálogo’ en el Museo de Kanazawa; ‘Forma apriorística’, en la Casa Flor y ‘Repetición de una situación topológica’, en el Centro Rolex. Todas esas estrategias, o juegos, mantienen aspectos comunes relativos a la forma arquitectónica, precisamente los aspectos Gifu: la repetición aplicada al patrón espacial, y lo que conlleva: desvinculación entre forma y función y la nueva acepción de flexibilidad. ‘Tableros de Juego’ consiste en configurar cada sistema de proyecto (estructura, cerramientos, particiones y mobiliario) eligiendo elementos ofrecidos por una geometría de base, en cada proyecto la suya, en general reticular: intersecciones, líneas, módulos. Cada sistema se configura, en principio, sin relación de subordinación con cualquiera de los demás; cuando esa subordinación es ineludible, el juego determina que el sistema portante no puede materializar el orden geométrico de base, lo que se traduce en que no ejerce el papel dominante. Por lo tanto, ‘Tableros de Juego’ transgrede la lógica de la planta libre moderna: la estructura ni refleja ni revela el orden de base y los sistemas no respetan las relaciones de subordinación jerárquica y encadenada que aquella determinaba. Esta estrategia de ‘Tableros de juego’ deriva en soluciones y proyectos formales muy distintos: los proyectos de Oogaki y Park Café, que presentarían ‘Tableros de Juego’ incipientes; De Kunstlinie en Almere y la Escuela de Zollverein, que presentarían una consolidación de esta estrategia; y el Pabellón de Vidrio de Toledo que resultaría de la subversión de la estrategia. Este último proyecto, además, lleva el concepto de repetición más allá del elemento constructivo y del patrón espacial (que en este caso tiene forma de burbuja) parar acabar afectando a la propia experiencia del espectador, que esté donde esté, siempre tiene la sensación de estar en el mismo sitio. Esta tesis denomina a ese espacio repetitivo como ‘espacio mantra’. La estrategia ‘Elementos de Catálogo’ se ilustra con el Museo de Kanazawa. Su lógica parte de la definición de una serie de elementos, muy pocos, y se basa en el ingente número de posibles combinaciones entre sí. Gifu habría anunciado el catalogo de elementos en la caracterización de sus patrones espaciales. La estrategia ‘Forma Apriorística’ se ilustra con la Casa Flor. La decisión sobre el tipo de forma -en este caso la de una ameba- estaría al principio del proceso proyectual, lo que no quiere decir que sea una forma arbitraria: la forma de la ameba lleva implícita la repetición de un patrón espacial (el seudópodo) y una apoteosis del concepto de repetición que, alcanzando la experiencia espacial, da lugar a un espacio repetitivo o mantra. El ‘Espacio Mantra’ es uno de los leitmotivs, que se emplean como argumento en la última estrategia formal que la Tesis decodifica: el Centro Rolex. Con respecto a la estructura portante, la tesis identifica y traza una trayectoria de cinco estrategias proyectuales: preeminencia, ocultación, disolución, desaparición y desvirtuación. --Ocultación, reduce el papel dominante de la estructura. Al principio es una ocultación literal, casi un tapado de los elementos estructurales, como en Gifu; luego se hace más sofisticada, como la ocultación por camuflaje o la paradójica ocultación por multiplicación de Park Café. --La disolución merma la condición dominante de la estructura que en lugar de configurarse como sistema unitario u homogéneo se fragmenta en varios subsistemas. --La desaparición se refiere a estructuras que desaparecen como sistemas propios y autónomos, a proyectos en los que la función portante es desempeñada por otros sistemas como el de las particiones. La desaparición culmina con la Casa Flor, cuyo perímetro ejerce la función portante y además es transparente, está desmaterializado: la estructura se ha hecho invisible, ha desaparecido. --La desvirtuación se refiere a estructuras que sí se presentan como sistemas propios y autónomos, pero dejan de tener un papel preeminente por cuanto no materializan el orden de base: esta estrategia es correlativa a la estrategia formal ‘Tableros de juego’. Las conclusiones de la tesis están en la propia organización de la tesis: la identificación de las estrategias. Aún así, y como epílogos, se exponen seis. Las dos primeras subrayan el hilo conductor del trabajo realizado, que radica en la cualidad genérica de las estrategias proyectuales en Sejima-SANAA. Las cuatro siguientes dilucidan hasta qué punto hay, en sus proyectos, rasgos o significantes formales y/o estructurales que sean a su vez señales características del panorama arquitectónico contemporáneo; y plantean la pregunta estrella: ¿hay algunos que, apuntando más lejos, supongan aportaciones originales? --Como aportaciones originales la tesis destaca: la identificación entre el ideal genérico y proyecto concreto; y la propuesta de un espacio nuevo, híbrido, una suerte de estadio intermedio entre el espacio subdividido y compartimentado de la tradición y el continuo moderno. --Como síntomas de contemporaneidad se destacan: respecto de la forma, la traslación de la especificidad formal de la parte al conjunto; y respecto de la estructura, la tendencia contemporánea a hacer estructuras cada vez más ligeras y livianas, que tienden a lo evanescente. Ésta última, la tendencia al evanescencia estructural, podría tener la condición de aportación original, no en vano la desaparición de la estructura lleva la evanescencia hacia sus últimas consecuencias, y en el caso de estructuras con presencia física, hace que dejen de ser el sistema ordenador orquestador del proceso proyectual. ABSTRACT The Thesis decodes a selection of twenty representative Sejima-SANAA projects, from the first one built, the Platform I House in 1987, to the Rolex Center in 2010, year in which Sejima and Nishizawa –SANAA- received the Pritzker Prize. Eleven projects are from Sejima: Platform I, Platform II, Saishunkan Seiyaku Women´s Dormitory, N- House, Pachinco Parlor I, Villa in the Forest, Policy Box at Chofu Station, Y-House, Gifu Kitigata Apartment, World City Expo ´96 Facilities Building, Pachinko Parlor III; and nine from SANAA: Multimedia Workshop in Ogaki, Metropolitan Housing Studies, Park Café in Koga, De Kunstlinie in Almere, Kanazawa Museum, Glass Pavilion at the Toledo Museum of Art, Zollverein School, Flower House and the Rolex Center. This decoding reads the Sejima-SANAA’s projects inversely aiming ‘to reconstruct', in a fictitious simulation exercise, a likely and coherent version of what her/their projectual processes ‘could’ have been; ‘could’, because the true ones are impossible to explain. The ones proposed here pretend only to be likely and reasonable. By so doing the Thesis tries to contribute to the understanding and comprehension of Sejima-SANAA architecture and, tangentially and to a lesser extent, to the theory of architectural projects exercise. Decoding centers in two specific aspects: architectural form, and projectual role of the load bearing structure. Both decodes inevitably extend to other related aspects such as, for example, the nature of space. The research procedure begun by carrying out an objective and detailed description of the formal and structural signifiers of each project; looking at them from their physical and geometric configuration. Taken to the limit, the ‘objective’ descriptions allowed the conceptual structures and underlying logics of each project to arise. Together with critical interpretations, which related and confronted them with other architectures and well-known projectual working ways, it became possible to outline and trace the intended fictitious reconstruction decodes. The descriptive analytical work materialized in twenty critical essays, and was accompanied by a set of other essays on subjects suggested or demanded by the research process. Together, all those texts were the material basis on which thesis work was built. Looking at the whole and taking it from there, the thesis identifies two related projectual trajectories: a trajectory of formal strategies and a trajectory of strategies having to do with structural systems and components. Both, together, constitute the bulk of the thesis, as presented in the four central chapters. Preceding them there is an introductory chapter outlining the biographical path of Kazuyo Sejima and the professional trajectory of Sejima-SANAA. And following them there is another one containing transversal texts on form, place and space. The thesis ends with a synthesis on conclusions. The formal strategies are displayed in three chapters. The first one, `Early formal strategies' groups the first phase projects by Sejima. The second one, ‘Formal strategies of Gifu’s paradigm’, is entirely dedicated to the Gifu apartments project, 1994-98, which according to this thesis meant an important inflexion point in Sejima’s trajectory; so much so that the third chapter is named `Formal strategies after Gifu' and gathers the selected projects that followed it. The ‘Early formal strategies', diverse and tentative, move in general around two well-known projectual composition methods ‘composition by parts’, and ‘systematic composition’. This last one –systematic composition- begins and leads in SANAA’s trajectory an aspect which will remain relevant from here on: the understanding of the project as if it were an specific instance of a generic proposal in which -below and beyond the project tangible reality- there lays a logic that could be applicable at other places, for other dimensions, even with other programs; from each project, other projects of the same family could rise. The set of projects using this systematic composition method include, among others, the ‘Platform II House, based on the definition of a constructive element and of rules having to do with its replicas and their possible groupings. It also includes the Saishunkan Seiyaku Women Residence -project that launched Sejima to international fame- that could also be seen as a system, but of a different kind: a system based on the regular repetition of a series of elements along a directive line, thus generating a hypothetical infinite container of which the project would be only a fragment. The formal strategy of the Gifu apartments building would push further towards the generic project concept, adopting the logic of a game. The project would be a bout, a round, one play…, but not the only possible one; others could be played. The thesis confirms this game hypothesis -after having formulated `The Game of Gifu' and identified its elements (board, chips, rules and procedures)- playing the one play from which the building as projected by Sejima would have raised. Gifu extends the concept of ‘repeating a constructive element’ to that of ‘repeating a space pattern element’, and to what it implies: the decoupling of form and function, leading to a new concept of flexibility that no longer refers to the flexible use of the constructed building but to the projectual moment at which the specific functions are assigned to the space patterns. This thesis proposes that this allocation of functions would be one of the last steps in projectual process, quite opposite from the modern premise: “form follows function”. The Formal strategies after Gifu do also have a game logic; but, as their names reveal, each strategy responds to a different game: ‘Game Boards’, present with different maturity levels in several projects; ‘Elements from a Catalogue’, in the Kanazawa Museum; ‘Aprioristic Form’, in the Flower House; and ‘Repetition of a topologic situation', in the Rolex Center. All of these strategies, or games, maintain common aspects having to do with architectural form; aspects that were already present, precisely, in Gifu: repetition of space pattern units, uncoupling of form and function, and a new meaning of flexibility. -`Game Boards’ consists on setting up a base geometry -each project his, generally reticular- and give form to each project system (structure, closings, partitions and furniture) by choosing elements -intersections, lines, modules- it offers. Each project system is formed, in principle, with no subordinated relation with any of the others; when subordination is unavoidable, the game rules determine that the load bearing structural system may not be the one to materialize the base geometric order, which means that it does not exert the dominant role. Therefore, ‘Game Boards' transgresses the Modern logic, because the structure neither reflects nor reveals the base order, and because the systems do not respect any of the hierarchic and chained subordination relations that the ‘free plan’ called for. ‘Game Boards' leads to quite different solutions and formal projects: the Oogaki and Park Coffee projects show incipient Game Boards; The Almere Kunstlinie and the Zollverein School present consolidations of this strategy; and the Toledo’s Glass Pavilion results from subverting the strategy. In addition, the Toledo project takes the repetition concept beyond that of using a constructive element and a space pattern element (in this case with a bubble form) to end up affecting the personal experience of the spectator, who, wherever he is, feels to always be in the same place. This thesis denominates that repetitive space as ‘Mantra space '. -‘Elements from a Catalogue’ is shown with the Kanazawa Museum. Its logic starts from the definition of a series of elements, very few, and it is based on the huge number of possible combinations among them. The ‘Elements from a Catalogue’ approach was announced in the Gifu project when characterizing its space pattern elements. -Aprioristic Form' is illustrated by the Flower House. The decision on the type of form -in this case the form of an amoeba- would be the beginning of the projectual process, but it does not mean it is arbitrary form: the amoeba form implies repeating a space pattern (pseudopodia) and an apotheosis of the repetition concept: embracing the space experience, it gives rise to a repetitive or mantra space. ‘Mantra Space’ is one of leitmotivs used as an argument in the last formal strategy Thesis decodes: the Rolex Center. With respect to the ‘Projectual strategies of the load bearing structure’, the thesis finds and traces a trajectory of five projectual strategies: ‘preeminence, concealment, dissolution, disappearance and desvirtuación’. --Preeminence is present in Sejima’s first works in which she resorts to structures which have a dominant preeminent role in the project in so far as they impersonate the greater scale and/or materialize the base geometric order. In later works that preeminence will be inverted, the projects aiming towards its opposite: lighter, slighter, smaller structures. -Concealment reduces the dominant role of the structure. At the outset concealment is literal, almost hiding the structural elements, as in Gifu; soon it will become more sophisticated, such as the concealment by camouflage or the paradoxical concealment by multiplication in the Koga Park Café. -Dissolution diminishes the dominant condition of the structure: instead of its’ being configured as unitary or homogenous system is fragmented in several subsystems. -Disappearance talks about structures that fade away as self referred and independent systems; projects in which the load bearing function is carried out by other systems such as the set of partitions. Disappearance reaches its zenith at the Flower House, whose perimeter functions structurally being, in addition, transparent, immaterial: its structure has become invisible, has disappeared. -Desvirtuación talks about structures that do appear like independent self-systems, but which that do not longer have a preeminent paper, inasmuch as they do not materialize the base order. This strategy correlates with the ‘Game Boards’ formal strategy. The thesis conclusions are show by the organization of the thesis itself: its identification of the different strategies. Even so, as epilogues, the thesis exposes six ‘Conclusions’. The first two emphasize the leading thread of the work done, rooted in the generic quality of the Sejima-SANAA projectual strategies. The following four expound to what extent their projects show features, or formal and/or structural signifiers, which also are or can be read as characteristic signals of the contemporary architectonic panorama, and raise the key question: aiming farther, may some of them be taken as original contributions? -As original contributions the conclusions highlight: the identification between the generic ideal and the concrete project; and the proposal of a new, hybrid space, kind of an intermediate stage between the traditional subdivided compartmented space and the continuous modern. -As symptoms of contemporaneousness: in relation to the form it highlights the transferring of the formal specificity from the part to the whole; and in relation to the structure, it underscore the contemporary tendency towards lighter and growingly slimmer structures, tending to the evanescent. This last one, the tendency towards structural evanescence, could have condition of being an original contribution, not in vain it carries the structural disappearance towards its last consequences; and in the case of structures with physical presence, it makes them to cease being the ordering system orchestrating the projectual process.