Nonviral Gene Delivery of Growth and Differentiation Factor 5 to Human Mesenchymal Stem Cells Injected into a 3D Bovine Intervertebral Disc Organ Culture System

Intervertebral disc (IVD) cell therapy with unconditioned 2D expanded mesenchymal stem cells (MSC) is a promising concept yet challenging to realize. Differentiation of MSCs by nonviral gene delivery of growth and differentiation factor 5 (GDF5) by electroporation mediated gene transfer could be an excellent source for cell transplantation. Human MSCs were harvested from bone marrow aspirate and GDF5 gene transfer was achieved by in vitro electroporation. Transfected cells were cultured as monolayers and as 3D cultures in 1.2% alginate bead culture. MSC expressed GDF5 efficiently for up to 21 days. The combination of GDF5 gene transfer and 3D culture in alginate showed an upregulation of aggrecan and SOX9, two markers for chondrogenesis, and KRT19 as a marker for discogenesis compared to untransfected cells. The cells encapsulated in alginate produced more proteoglycans expressed in GAG/DNA ratio. Furthermore, GDF5 transfected MCS injected into an IVD papain degeneration organ culture model showed a partial recovery of the GAG/DNA ratio after 7 days. In this study we demonstrate the potential of GDF5 transfected MSC as a promising approach for clinical translation for disc regeneration.

3D Conversion using Vanishing Points and Image Warping

We describe a user assisted technique for 3D stereo conversion from 2D images. Our approach exploits the geometric structure of perspective images including vanishing points. We allow a user to indicate lines, planes, and vanishing points in the input image, and directly employ these as constraints in an image warping framework to produce a stereo pair. By sidestepping explicit construction of a depth map, our approach is applicable to more general scenes and avoids potential artifacts of depth-image-based rendering. Our method is most suitable for scenes with large scale structures such as buildings.

Quantitative 3D strain analysis in analogue experiments: Integration of X-ray computed tomography and digital volume correlation techniques

The combination of scaled analogue experiments, material mechanics, X-ray computed tomography (XRCT) and Digital Volume Correlation techniques (DVC) is a powerful new tool not only to examine the 3 dimensional structure and kinematic evolution of complex deformation structures in scaled analogue experiments, but also to fully quantify their spatial strain distribution and complete strain history. Digital image correlation (DIC) is an important advance in quantitative physical modelling and helps to understand non-linear deformation processes. Optical non-intrusive (DIC) techniques enable the quantification of localised and distributed deformation in analogue experiments based either on images taken through transparent sidewalls (2D DIC) or on surface views (3D DIC). X-ray computed tomography (XRCT) analysis permits the non-destructive visualisation of the internal structure and kinematic evolution of scaled analogue experiments simulating tectonic evolution of complex geological structures. The combination of XRCT sectional image data of analogue experiments with 2D DIC only allows quantification of 2D displacement and strain components in section direction. This completely omits the potential of CT experiments for full 3D strain analysis of complex, non-cylindrical deformation structures. In this study, we apply digital volume correlation (DVC) techniques on XRCT scan data of “solid” analogue experiments to fully quantify the internal displacement and strain in 3 dimensions over time. Our first results indicate that the application of DVC techniques on XRCT volume data can successfully be used to quantify the 3D spatial and temporal strain patterns inside analogue experiments. We demonstrate the potential of combining DVC techniques and XRCT volume imaging for 3D strain analysis of a contractional experiment simulating the development of a non-cylindrical pop-up structure. Furthermore, we discuss various options for optimisation of granular materials, pattern generation, and data acquisition for increased resolution and accuracy of the strain results. Three-dimensional strain analysis of analogue models is of particular interest for geological and seismic interpretations of complex, non-cylindrical geological structures. The volume strain data enable the analysis of the large-scale and small-scale strain history of geological structures.

Clinical Significance of 3D Ultrasound Compared to MRI in Uterine Malformations

Purpose: Congenital uterine anomalies often remain asymptomatic until they cause problems, for example during pregnancy. We studied the diagnostic aspects of two- and three-dimensional ultrasound and MRI. Materials and Methods: 63 women referred for suspected uterine anomalies were studied: In the first group (until July 2008) with 2 D-US and MRI, in the second group (from August 2008) additionally with 3 D-US; these women also had diagnostic or therapeutic operative confirmation. In the third group, only 3D-US was used. Results: In all women 3D-US was possible and successful. The most common anomaly was a subseptate uterus, while a septate uterus was less frequent, and uterus bicornis (unicollis) and uterus didelphys (bicornis bicollis) were rare. The women in the first two groups all underwent at least diagnostic hysteroscopy, and some (subseptate or septate uterus) underwent operative hysteroscopy. After preoperative volume imaging, laparoscopies were required less often. 3D-US diagnoses as judged by intraoperative findings were correct in 100 % of cases, while the MRI diagnoses in the same group were correct in only 7/13 cases. Conclusion: Since the introduction of volume imaging (MRI, later 3 D-US), laparoscopy during hysteroscopic septum resection was not necessary in the majority of cases. 3D-US brings the diagnostics of uterine anomalies back into the hands of the gynecologist and can provide the gynecological surgeon with a higher subjective degree of certainty during operative hysteroscopy.

Development and characterization of a scaffold-free 3D spheroid model of iPSC-derived human cardiomyocytes

Purpose: Cardiomyocytes are terminally differentiated cells in the adult heart and ischemia and cardiotoxic compounds can lead to cell death and irreversible decline of cardiac function. As testing platforms, isolated organs and primary cells from rodents have been the standard in research and toxicology, but there is a need for better models that more faithfully recapitulate native human biology. Hence, a new in vitro model comprising the advantages of 3D cell culture and the availability of induced pluripotent stem cells (iPSC) from human origin was developed and characterized. Methods: Human cardiomyocytes (CMs) derived from induced pluripotent stem cells (iPSCs) were studied in standard 2D culture and as cardiac microtissues (MTs) formed in hanging drops. 2D cultures were examined using immunofluorescence microscopy and Western blotting while the cardiac MTs were subjected to immunofluorescence, contractility, and pharmacological investigations. Results: iPSC-derived CMs in 2D culture showed well-formed myofibrils, cell-cell contacts positive for connexin-43, and other typical cardiac proteins. The cells reacted to pro-hypertrophic growth factors with a substantial increase in myofibrils and sarcomeric proteins. In hanging drop cultures, iPSC-derived cardiomyocytes formed spheroidal MTs within 4 days showing a homogeneous tissue structure with well-developed myofibrils extending throughout the whole spheroid without a necrotic core. MTs showed spontaneous contractions for more than 4 weeks that were recorded by optical motion tracking, sensitive to temperature, and responsive to electrical pacing. Contractile pharmacology was tested with several agents known to modulate cardiac rate and viability. Calcium-transients underlay the contractile activity and were also responsive to electrical stimulation, caffeine-induced Ca2+-release, extracellular calcium levels. Conclusions: 3D culture using iPSC-derived human cardiomyocytes provides an organoid human-based cellular platform that is free of necrosis and recapitulates vital cardiac functionality, thereby providing new and promising relevant model for the evaluation and development of new therapies and detection of cardiotoxicity.

VIRTOPSY--scientific documentation, reconstruction and animation in forensic: individual and real 3D data based geo-metric approach including optical body/object surface and radiological CT/MRI scanning.

Until today, most of the documentation of forensic relevant medical findings is limited to traditional 2D photography, 2D conventional radiographs, sketches and verbal description. There are still some limitations of the classic documentation in forensic science especially if a 3D documentation is necessary. The goal of this paper is to demonstrate new 3D real data based geo-metric technology approaches. This paper present approaches to a 3D geo-metric documentation of injuries on the body surface and internal injuries in the living and deceased cases. Using modern imaging methods such as photogrammetry, optical surface and radiological CT/MRI scanning in combination it could be demonstrated that a real, full 3D data based individual documentation of the body surface and internal structures is possible in a non-invasive and non-destructive manner. Using the data merging/fusing and animation possibilities, it is possible to answer reconstructive questions of the dynamic development of patterned injuries (morphologic imprints) and to evaluate the possibility, that they are matchable or linkable to suspected injury-causing instruments. For the first time, to our knowledge, the method of optical and radiological 3D scanning was used to document the forensic relevant injuries of human body in combination with vehicle damages. By this complementary documentation approach, individual forensic real data based analysis and animation were possible linking body injuries to vehicle deformations or damages. These data allow conclusions to be drawn for automobile accident research, optimization of vehicle safety (pedestrian and passenger) and for further development of crash dummies. Real 3D data based documentation opens a new horizon for scientific reconstruction and animation by bringing added value and a real quality improvement in forensic science.

Stereoselective virtual screening of the ZINC database using atom pair 3D-fingerprints

Background Tools to explore large compound databases in search for analogs of query molecules provide a strategically important support in drug discovery to help identify available analogs of any given reference or hit compound by ligand based virtual screening (LBVS). We recently showed that large databases can be formatted for very fast searching with various 2D-fingerprints using the city-block distance as similarity measure, in particular a 2D-atom pair fingerprint (APfp) and the related category extended atom pair fingerprint (Xfp) which efficiently encode molecular shape and pharmacophores, but do not perceive stereochemistry. Here we investigated related 3D-atom pair fingerprints to enable rapid stereoselective searches in the ZINC database (23.2 million 3D structures). Results Molecular fingerprints counting atom pairs at increasing through-space distance intervals were designed using either all atoms (16-bit 3DAPfp) or different atom categories (80-bit 3DXfp). These 3D-fingerprints retrieved molecular shape and pharmacophore analogs (defined by OpenEye ROCS scoring functions) of 110,000 compounds from the Cambridge Structural Database with equal or better accuracy than the 2D-fingerprints APfp and Xfp, and showed comparable performance in recovering actives from decoys in the DUD database. LBVS by 3DXfp or 3DAPfp similarity was stereoselective and gave very different analogs when starting from different diastereomers of the same chiral drug. Results were also different from LBVS with the parent 2D-fingerprints Xfp or APfp. 3D- and 2D-fingerprints also gave very different results in LBVS of folded molecules where through-space distances between atom pairs are much shorter than topological distances. Conclusions 3DAPfp and 3DXfp are suitable for stereoselective searches for shape and pharmacophore analogs of query molecules in large databases. Web-browsers for searching ZINC by 3DAPfp and 3DXfp similarity are accessible at www.gdb.unibe.ch webcite and should provide useful assistance to drug discovery projects.

Automated 3D Lumbar Intervertebral Disc Segmentation from MRI Data Sets

This paper proposed an automated three-dimensional (3D) lumbar intervertebral disc (IVD) segmentation strategy from Magnetic Resonance Imaging (MRI) data. Starting from two user supplied landmarks, the geometrical parameters of all lumbar vertebral bodies and intervertebral discs are automatically extracted from a mid-sagittal slice using a graphical model based template matching approach. Based on the estimated two-dimensional (2D) geometrical parameters, a 3D variable-radius soft tube model of the lumbar spine column is built by model fitting to the 3D data volume. Taking the geometrical information from the 3D lumbar spine column as constraints and segmentation initialization, the disc segmentation is achieved by a multi-kernel diffeomorphic registration between a 3D template of the disc and the observed MRI data. Experiments on 15 patient data sets showed the robustness and the accuracy of the proposed algorithm.

Comparison of 2.5D and 3D Quantification of Femoral Head Coverage in Normal Control Subjects and Patients with Hip Dysplasia.

Hip dysplasia is characterized by insufficient femoral head coverage (FHC). Quantification of FHC is of importance as the underlying goal of the surgery to treat hip dysplasia is to restore a normal acetabular morphology and thereby to improve FHC. Unlike a pure 2D X-ray radiograph-based measurement method or a pure 3D CT-based measurement method, previously we presented a 2.5D method to quantify FHC from a single anteriorposterior (AP) pelvic radiograph. In this study, we first quantified and compared 3D FHC between a normal control group and a patient group using a CT-based measurement method. Taking the CT-based 3D measurements of FHC as the gold standard, we further quantified the bias, precision and correlation between the 2.5D measurements and the 3D measurements on both the control group and the patient group. Based on digitally reconstructed radiographs (DRRs), we investigated the influence of the pelvic tilt on the 2.5D measurements of FHC. The intraclass correlation coefficients (ICCs) for absolute agreement was used to quantify interobserver reliability and intraobserver reproducibility of the 2.5D measurement technique. The Pearson correlation coefficient, r, was used to determine the strength of the linear association between the 2.5D and the 3D measurements. Student's t-test was used to determine whether the differences between different measurements were statistically significant. Our experimental results demonstrated that both the interobserver reliability and the intraobserver reproducibility of the 2.5D measurement technique were very good (ICCs > 0.8). Regression analysis indicated that the correlation was very strong between the 2.5D and the 3D measurements (r = 0.89, p < 0.001). Student's t-test showed that there were no statistically significant differences between the 2.5D and the 3D measurements of FHC on the patient group (p > 0.05). The results of this study provided convincing evidence demonstrating the validity of the 2.5D measurements of FHC from a single AP pelvic radiograph and proved that it could serve as a surrogate for 3D CT-based measurements. Thus it may be possible to use this method to avoid a CT scan for the purpose of estimating 3D FHC in diagnosis and post-operative treatment evaluation of patients with hip dysplasia.

Accuracy of linear measurements using three imaging modalities: two lateral cephalograms and one 3D model from CBCT data

BACKGROUND The aim of this study was to evaluate the accuracy of linear measurements on three imaging modalities: lateral cephalograms from a cephalometric machine with a 3 m source-to-mid-sagittal-plane distance (SMD), from a machine with 1.5 m SMD and 3D models from cone-beam computed tomography (CBCT) data. METHODS Twenty-one dry human skulls were used. Lateral cephalograms were taken, using two cephalometric devices: one with a 3 m SMD and one with a 1.5 m SMD. CBCT scans were taken by 3D Accuitomo® 170, and 3D surface models were created in Maxilim® software. Thirteen linear measurements were completed twice by two observers with a 4 week interval. Direct physical measurements by a digital calliper were defined as the gold standard. Statistical analysis was performed. RESULTS Nasion-Point A was significantly different from the gold standard in all methods. More statistically significant differences were found on the measurements of the 3 m SMD cephalograms in comparison to the other methods. Intra- and inter-observer agreement based on 3D measurements was slightly better than others. LIMITATIONS Dry human skulls without soft tissues were used. Therefore, the results have to be interpreted with caution, as they do not fully represent clinical conditions. CONCLUSIONS 3D measurements resulted in a better observer agreement. The accuracy of the measurements based on CBCT and 1.5 m SMD cephalogram was better than a 3 m SMD cephalogram. These findings demonstrated the linear measurements accuracy and reliability of 3D measurements based on CBCT data when compared to 2D techniques. Future studies should focus on the implementation of 3D cephalometry in clinical practice.

Optimization of a label-free biosensor vertically characterized based on a periodic lattice of high aspect ratio SU-8 nano-pillars with a simplified 2D theoretical model

We have recently demonstrated a biosensor based on a lattice of SU8 pillars on a 1 μm SiO2/Si wafer by measuring vertically reflectivity as a function of wavelength. The biodetection has been proven with the combination of Bovine Serum Albumin (BSA) protein and its antibody (antiBSA). A BSA layer is attached to the pillars; the biorecognition of antiBSA involves a shift in the reflectivity curve, related with the concentration of antiBSA. A detection limit in the order of 2 ng/ml is achieved for a rhombic lattice of pillars with a lattice parameter (a) of 800 nm, a height (h) of 420 nm and a diameter(d) of 200 nm. These results correlate with calculations using 3D-finite difference time domain method. A 2D simplified model is proposed, consisting of a multilayer model where the pillars are turned into a 420 nm layer with an effective refractive index obtained by using Beam Propagation Method (BPM) algorithm. Results provided by this model are in good correlation with experimental data, reaching a reduction in time from one day to 15 minutes, giving a fast but accurate tool to optimize the design and maximizing sensitivity, and allows analyzing the influence of different variables (diameter, height and lattice parameter). Sensitivity is obtained for a variety of configurations, reaching a limit of detection under 1 ng/ml. Optimum design is not only chosen because of its sensitivity but also its feasibility, both from fabrication (limited by aspect ratio and proximity of the pillars) and fluidic point of view. (© 2011 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim)

Validación de dos algoritmos para la localización 3D del instrumental laparoscópico a partir de la imagen de vídeo endoscópico

Uno de los aspectos fundamentales en un sistema de cirugía guiada por imagen (CGI) es la localización del instrumental quirúrgico con respecto a la anatomía del paciente. Los sistemas basados en sensores ofrecen buenos niveles de precisión, pero son sensibles a distintas fuentes de ruido en el quirófano y contribuyen a la sobrecarga tecnológica del mismo. Una alternativa novedosa es analizar la imagen del vídeo endoscópico para llevar a cabo la detección y localización espacial del instrumental. Se presenta en este trabajo la validación de dos métodos, basados en el diámetro aparente y en la sección transversal del instrumental, para la localización espacial del instrumental a partir de los bordes y la posición 2D de la punta en la imagen. La validación, llevada a cabo en un simulador físico, se realiza comparando los resultados con el sistema Kinescan/IBV. Los resultados muestran para cada método un error medio de 12,7 y 12,8 mm respectivamente. La incorporación de estos algoritmos dentro del paradigma de navegación propuesto en el proyecto THEMIS permitirá al cirujano conocer la posición del instrumental de forma no intrusiva y transparente, sin necesidad de equipamiento adicional en el quirófano.

Posibilidades del motor 3D de la Nintendo DS

La consola portátil Nintendo DS es una plataforma de desarrollo muy presente entre la comunidad de desarrolladores independientes, con una extensa y nutrida escena homebrew. Si bien las capacidades 2D de la consola están muy aprovechadas, dado que la mayor parte de los esfuerzos de los creadores amateur están enfocados en este aspecto, el motor 3D de ésta (el que se encarga de representar en pantalla modelos tridimensionales) no lo está de igual manera. Por lo tanto, en este proyecto se tiene en vista determinar las capacidades gráficas de la Nintendo DS. Para ello se ha realizado una biblioteca de funciones en C que permite aprovechar las posibilidades que ofrece la consola en el terreno 3D y que sirve como herramienta para la comunidad homebrew para crear aplicaciones 3D de forma sencilla, dado que se ha diseñado como un sistema modular y accesible. En cuanto al proceso de renderizado se han sacado varias conclusiones. En primer lugar se ha determinado la posibilidad de asignar varias componentes de color a un mismo vértice (color material reactivo a la iluminación, color por vértice directo y color de textura), tanto de forma independiente como simultáneamente, pudiéndose utilizar para aplicar diversos efectos al modelo, como iluminación pre-calculada o simulación de una textura mediante color por vértice, ahorrando en memoria de video. Por otro lado se ha implementado un sistema de renderizado multi-capa, que permite realizar varias pasadas de render, pudiendo, de esta forma, aplicar al modelo una segunda textura mezclada con la principal o realizar un efecto de reflexión esférica. Uno de los principales avances de esta herramienta con respecto a otras existentes se encuentra en el apartado de animación. El renderizador desarrollado permite por un lado animación por transformación, consistente en la animación de mallas o grupos de vértices del modelo mediante el movimiento de una articulación asociada que determina su posición y rotación en cada frame de animación. Por otro lado se ha implementado un sistema de animación por muestreo de vértices mediante el cual se determina la posición de éstos en cada instante de la animación, generando frame a frame las poses que componen el movimiento (siendo este último método necesario cuando no se puede animar una malla por transformación). Un mismo modelo puede contener diferentes esqueletos, animados independientemente entre sí, y cada uno de ellos tener definidas varias costumbres de animación que correspondan a movimientos contextuales diferentes (andar, correr, saltar, etc). Además, el sistema permite extraer cualquier articulación para asociar su transformación a un objeto estático externo y que éste siga el movimiento de la animación, pudiendo así, por ejemplo, equipar un objeto en la mano de un personaje. Finalmente se han implementado varios efectos útiles en la creación de escenas tridimensionales, como el billboarding (tanto esférico como cilíndrico), que restringe la rotación de un modelo para que éste siempre mire a cámara y así poder emular la apariencia de un objeto tridimensional mediante una imagen plana, ahorrando geometría, o emplearlo para realizar efectos de partículas. Por otra parte se ha implementado un sistema de animación de texturas por subimágenes que permite generar efectos de movimiento mediante imágenes, sin necesidad de transformar geometría. ABSTRACT. The Nintendo DS portable console has received great interest within the independent developers’ community, with a huge homebrew scene. The 2D capabilities of this console are well known and used since most efforts of the amateur creators has been focused on this point. However its 3D engine (which handles with the representation of three-dimensional models) is not equally used. Therefore, in this project the main objective is to assess the Nintendo DS graphic capabilities. For this purpose, a library of functions in C programming language has been coded. This library allows the programmer to take advantage of the possibilities that the 3D area brings. This way the library can be used by the homebrew community as a tool to create 3D applications in an easy way, since it has been designed as a modular and accessible system. Regarding the render process, some conclusions have been drawn. First, it is possible to assign several colour components to the same vertex (material colour, reactive to the illumination, colour per vertex and texture colour), independently and simultaneously. This feature can be useful to apply certain effects on the model, such as pre-calculated illumination or the simulation of a texture using colour per vertex, providing video memory saving. Moreover, a multi-layer render system has been implemented. This system allows the programmer to issue several render passes on the same model. This new feature brings the possibility to apply to the model a second texture blended with the main one or simulate a spherical reflection effect. One of the main advances of this tool over existing ones consists of its animation system. The developed renderer includes, on the one hand, transform animation, which consists on animating a mesh or groups of vertices of the model by the movement of an associated joint. This joint determines position and rotation of the mesh at each frame of the animation. On the other hand, this tool also implements an animation system by vertex sampling, where the position of vertices is determined at every instant of the animation, generating the poses that build up the movement (the latter method is mandatory when a mesh cannot be animated by transform). A model can contain multiple skeletons, animated independently, each of them being defined with several animation customs, corresponding to different contextual movements (walk, run, jump, etc). Besides, the system allows extraction of information from any joint in order to associate its transform to a static external object, which will follow the movement of the animation. This way, any object could be equipped, for example, on the hand of a character. Finally, some useful effects for the creation of three-dimensional scenes have been implemented. These effects include billboarding (both spherical and cylindrical), which constraints the rotation of a model so it always looks on the camera's direction. This feature can provide the ability to emulate the appearance of a three-dimensional model through a flat image (saving geometry). It can also be helpful in the implementation of particle effects. Moreover, a texture animation system using sub-images has also been implemented. This system allows the generation of movement by using images as textures, without having to transform geometry.

3D Videoconferencing system using spatial scalability

An implementation of a real-time 3D videoconferencing system using the currently available technology is presented. This appr oach is based on the side by side spatial compression of the stereoscopic images . The encoder and the decoder have b een implemented in a standard personal computer and a conventional 3D comp atible TV has been used to present the frames. Moreover, the users without 3D technology can use the system because 2D compatibility mode has been implemented in the decoder. The performance res ults show that a conventional computer can be used for encod ing/decoding audio and video streams and the delay in the transmission is lower than 200 ms.

Bayesian scene analysis for multi-camera 3D tracking and camera positioning

Los sistemas de seguimiento mono-cámara han demostrado su notable capacidad para el análisis de trajectorias de objectos móviles y para monitorización de escenas de interés; sin embargo, tanto su robustez como sus posibilidades en cuanto a comprensión semántica de la escena están fuertemente limitadas por su naturaleza local y monocular, lo que los hace insuficientes para aplicaciones realistas de videovigilancia. El objetivo de esta tesis es la extensión de las posibilidades de los sistemas de seguimiento de objetos móviles para lograr un mayor grado de robustez y comprensión de la escena. La extensión propuesta se divide en dos direcciones separadas. La primera puede considerarse local, ya que está orientada a la mejora y enriquecimiento de las posiciones estimadas para los objetos móviles observados directamente por las cámaras del sistema; dicha extensión se logra mediante el desarrollo de un sistema multi-cámara de seguimiento 3D, capaz de proporcionar consistentemente las posiciones 3D de múltiples objetos a partir de las observaciones capturadas por un conjunto de sensores calibrados y con campos de visión solapados. La segunda extensión puede considerarse global, dado que su objetivo consiste en proporcionar un contexto global para relacionar las observaciones locales realizadas por una cámara con una escena de mucho mayor tamaño; para ello se propone un sistema automático de localización de cámaras basado en las trayectorias observadas de varios objetos móviles y en un mapa esquemático de la escena global monitorizada. Ambas líneas de investigación se tratan utilizando, como marco común, técnicas de estimación bayesiana: esta elección está justificada por la versatilidad y flexibilidad proporcionada por dicho marco estadístico, que permite la combinación natural de múltiples fuentes de información sobre los parámetros a estimar, así como un tratamiento riguroso de la incertidumbre asociada a las mismas mediante la inclusión de modelos de observación específicamente diseñados. Además, el marco seleccionado abre grandes posibilidades operacionales, puesto que permite la creación de diferentes métodos numéricos adaptados a las necesidades y características específicas de distintos problemas tratados. El sistema de seguimiento 3D con múltiples cámaras propuesto está específicamente diseñado para permitir descripciones esquemáticas de las medidas realizadas individualmente por cada una de las cámaras del sistema: esta elección de diseño, por tanto, no asume ningún algoritmo específico de detección o seguimiento 2D en ninguno de los sensores de la red, y hace que el sistema propuesto sea aplicable a redes reales de vigilancia con capacidades limitadas tanto en términos de procesamiento como de transmision. La combinación robusta de las observaciones capturadas individualmente por las cámaras, ruidosas, incompletas y probablemente contaminadas por falsas detecciones, se basa en un metodo de asociación bayesiana basado en geometría y color: los resultados de dicha asociación permiten el seguimiento 3D de los objetos de la escena mediante el uso de un filtro de partículas. El sistema de fusión de observaciones propuesto tiene, como principales características, una gran precisión en términos de localización 3D de objetos, y una destacable capacidad de recuperación tras eventuales errores debidos a un número insuficiente de datos de entrada. El sistema automático de localización de cámaras se basa en la observación de múltiples objetos móviles y un mapa esquemático de las áreas transitables del entorno monitorizado para inferir la posición absoluta de dicho sensor. Para este propósito, se propone un novedoso marco bayesiano que combina modelos dinámicos inducidos por el mapa en los objetos móviles presentes en la escena con las trayectorias observadas por la cámara, lo que representa un enfoque nunca utilizado en la literatura existente. El sistema de localización se divide en dos sub-tareas diferenciadas, debido a que cada una de estas tareas requiere del diseño de algoritmos específicos de muestreo para explotar en profundidad las características del marco desarrollado: por un lado, análisis de la ambigüedad del caso específicamente tratado y estimación aproximada de la localización de la cámara, y por otro, refinado de la localización de la cámara. El sistema completo, diseñado y probado para el caso específico de localización de cámaras en entornos de tráfico urbano, podría tener aplicación también en otros entornos y sensores de diferentes modalidades tras ciertas adaptaciones. ABSTRACT Mono-camera tracking systems have proved their capabilities for moving object trajectory analysis and scene monitoring, but their robustness and semantic possibilities are strongly limited by their local and monocular nature and are often insufficient for realistic surveillance applications. This thesis is aimed at extending the possibilities of moving object tracking systems to a higher level of scene understanding. The proposed extension comprises two separate directions. The first one is local, since is aimed at enriching the inferred positions of the moving objects within the area of the monitored scene directly covered by the cameras of the system; this task is achieved through the development of a multi-camera system for robust 3D tracking, able to provide 3D tracking information of multiple simultaneous moving objects from the observations reported by a set of calibrated cameras with semi-overlapping fields of view. The second extension is global, as is aimed at providing local observations performed within the field of view of one camera with a global context relating them to a much larger scene; to this end, an automatic camera positioning system relying only on observed object trajectories and a scene map is designed. The two lines of research in this thesis are addressed using Bayesian estimation as a general unifying framework. Its suitability for these two applications is justified by the flexibility and versatility of that stochastic framework, which allows the combination of multiple sources of information about the parameters to estimate in a natural and elegant way, addressing at the same time the uncertainty associated to those sources through the inclusion of models designed to this end. In addition, it opens multiple possibilities for the creation of different numerical methods for achieving satisfactory and efficient practical solutions to each addressed application. The proposed multi-camera 3D tracking method is specifically designed to work on schematic descriptions of the observations performed by each camera of the system: this choice allows the use of unspecific off-the-shelf 2D detection and/or tracking subsystems running independently at each sensor, and makes the proposal suitable for real surveillance networks with moderate computational and transmission capabilities. The robust combination of such noisy, incomplete and possibly unreliable schematic descriptors relies on a Bayesian association method, based on geometry and color, whose results allow the tracking of the targets in the scene with a particle filter. The main features exhibited by the proposal are, first, a remarkable accuracy in terms of target 3D positioning, and second, a great recovery ability after tracking losses due to insufficient input data. The proposed system for visual-based camera self-positioning uses the observations of moving objects and a schematic map of the passable areas of the environment to infer the absolute sensor position. To this end, a new Bayesian framework combining trajectory observations and map-induced dynamic models for moving objects is designed, which represents an approach to camera positioning never addressed before in the literature. This task is divided into two different sub-tasks, setting ambiguity analysis and approximate position estimation, on the one hand, and position refining, on the other, since they require the design of specific sampling algorithms to correctly exploit the discriminative features of the developed framework. This system, designed for camera positioning and demonstrated in urban traffic environments, can also be applied to different environments and sensors of other modalities after certain required adaptations.