A presença que tem hoje a Medicina Nuclear como modalidade de obtenção de imagens médicas é muito importante e um dos principais procedimentos utilizados hoje nos centros de saúde, tendo como grande vantagem a capacidade de conseguir analisar o comportamento metabólico do paciente, fazendo possíveis diagnósticos precoces. Este projeto está baseado em imagens médicas obtidas através da modalidade PET (Positron Emission Tomography) a qual está tendo uma crescente difusão e aceitação. Para isso, temos desenvolvido uma estrutura integral de processamento de imagens tridimensionais PET, a qual está constituída por etapas consecutivas que se iniciam na obtenção das imagens padrões (gold standard), sendo utilizados volumes simulados ou phantoms do Ventrículo Esquerdo do Coração criadas como parte do projeto, assim como geradas a partir do software NCAT-4D. A seguir, nos volumes simulados, é introduzido ruído quântico tipo Poisson que é o ruído característico das imagens PET e feita uma análise que busca certificar que o ruído utilizado corresponde efetivamente ao ruído Poisson. Em sequência é executada a parte de pré-processamento, utilizando para este fim, um conjunto de filtros tais como o filtro da mediana, o filtro da Gaussiana ponderada e o filtro que mistura os conceitos da Transformada de Anscombe e o filtro pontual de Wiener. Posteriormente é aplicada a etapa de segmentação que é considerada a parte central da sequência de processamento. O processo de segmentação é baseado na teoria de Conectividade Fuzzy e para isso temos implementado quatro diferentes abordagens: Algoritmo Genérico, Algoritmo LIFO, Algoritmo kTetaFOEMS e o Algoritmo utilizando Pesos Dinâmicos. Sendo que os três primeiros algoritmos utilizam pesos específicos selecionados pelo usuário, foi preciso efetuar uma análise para determinar os melhores pesos de segmentação que se reflitam numa segmentação mais eficiente. Finalmente, para terminar a estrutura de processamento, um procedimento de avaliação foi utilizado como métrica para obter quantitativamente três parâmetros (Verdadeiro Positivo, Falso Positivo e Máxima Distância) que permitiram conhecer o nível de eficiência e precisão de nosso processo e do projeto em geral. Constatamos que os algoritmos implementados (filtros e algoritmos de segmentação) são bastante robustos e atingem ótimos resultados chegando-se a obter, para o caso do volume do Ventrículo Esquerdo simulado, taxas de VP e FP na ordem de 98.49 ± 0.27% e 2,19 ± 0.19%, respectivamente. Com o conjunto de procedimentos e escolhas feitas ao longo da estrutura de processamento, encerramos o projeto com a análise de um grupo de volumes produto de um exame PET real, obtendo a quantificação destes volumes. / The usefulness of Nuclear medicine nowadays as a modality to obtain medical images is very important, and it has turned into one of the main procedures utilized in Health Care Centers. Its great advantage is to analyze the metabolic behavior of the patient, by allowing early diagnosis. This project is based on medical images obtained by the PET modality (Positron Emission Tomography), which has won wide acceptance. Thus, we have developed an integral framework for processing Nuclear Medicine three-dimensional images of the PET modality, which is composed of consecutive steps that start with the generation of standard images (gold standard) by using simulated images or phantoms of the Left Ventricular Heart that were generated in this project, such as the ones obtained from the NCAT-4D software. Then Poisson quantum noise is introduced into the whole volume to simulate the characteristic noises in PET images and an analysis is performed in order to certify that the utilized noise is the Poisson noise effectively. Subsequently, the pre-processing is executed by using specific filters, such as the median filter, the weighted Gaussian filter, and the filter that joins the concepts of Anscombe Transformation and the Wiener filter. Then the segmentation, which is considered the most important and central part of the whole process, is implemented. The segmentation process is based on the Fuzzy Connectedness theory and for that purpose four different approaches were implemented: Generic algorithm, LIFO algorithm, kTetaFOEMS algorithm, and Dynamic Weight algorithm. Since the first three algorithms used specific weights that were selected by the user, an extra analysis was performed to determine the best segmentation constants that would reflect an accurate segmentation. Finally, at the end of the processing structure, an assessment procedure was used as a measurement tool to quantify some parameters that determined the level of efficiency and precision of our process and project. We have verified that the implemented algorithms (filters and segmentation algorithms) are fairly robust and achieve optimal results, assist to obtain, in the case of the Left Ventricular simulated, TP and FP rates in the order of 98.49 ± 0.27% and 2.19 ± 0.19%, respectively. With the set of procedures and choices made along of the processing structure, the project was concluded with the analysis of a volumes group from a real PET exam, obtaining the quantification of the volumes.

Environ 150.000 personnes souffrent en France d'une épilepsie partielle réfractaire à tous les médicaments. La chirurgie, qui constitue aujourd’hui le meilleur recours thérapeutique nécessite un bilan préopératoire complexe. L'analyse de données d'imagerie telles que l’imagerie par résonance magnétique (IRM) anatomique et la tomographie d’émission de positons (TEP) au FDG (fluorodéoxyglucose) tend à prendre une place croissante dans ce protocole, et pourrait à terme limiter de recourir à l’électroencéphalographie intracérébrale (SEEG), procédure très invasive mais qui constitue encore la technique de référence. Pour assister les cliniciens dans leur tâche diagnostique, nous avons développé un système d'aide au diagnostic (CAD) reposant sur l'analyse multivariée de données d'imagerie. Compte tenu de la difficulté relative à la constitution de bases de données annotées et équilibrées entre classes, notre première contribution a été de placer l'étude dans le cadre méthodologique de la détection du changement. L'algorithme du séparateur à vaste marge adapté à ce cadre là (OC-SVM) a été utilisé pour apprendre, à partir de cartes multi-paramétriques extraites d'IRM T1 de sujets normaux, un modèle prédictif caractérisant la normalité à l'échelle du voxel. Le modèle permet ensuite de faire ressortir, dans les images de patients, les zones cérébrales suspectes s'écartant de cette normalité. Les performances du système ont été évaluées sur des lésions simulées ainsi que sur une base de données de patients. Trois extensions ont ensuite été proposées. D'abord un nouveau schéma de détection plus robuste à la présence de bruit d'étiquetage dans la base de données d'apprentissage. Ensuite, une stratégie de fusion optimale permettant la combinaison de plusieurs classifieurs OC-SVM associés chacun à une séquence IRM. Enfin, une généralisation de l'algorithme de détection d'anomalies permettant la conversion de la sortie du CAD en probabilité, offrant ainsi une meilleure interprétation de la sortie du système et son intégration dans le bilan pré-opératoire global. / One third of patients suffering from epilepsy are resistant to medication. For these patients, surgical removal of the epileptogenic zone offers the possibility of a cure. Surgery success relies heavily on the accurate localization of the epileptogenic zone. The analysis of neuroimaging data such as magnetic resonance imaging (MRI) and positron emission tomography (PET) is increasingly used in the pre-surgical work-up of patients and may offer an alternative to the invasive reference of Stereo-electro-encephalo -graphy (SEEG) monitoring. To assist clinicians in screening these lesions, we developed a computer aided diagnosis system (CAD) based on a multivariate data analysis approach. Our first contribution was to formulate the problem of epileptogenic lesion detection as an outlier detection problem. The main motivation for this formulation was to avoid the dependence on labelled data and the class imbalance inherent to this detection task. The proposed system builds upon the one class support vector machines (OC-SVM) classifier. OC-SVM was trained using features extracted from MRI scans of healthy control subjects, allowing a voxelwise assessment of the deviation of a test subject pattern from the learned patterns. System performance was evaluated using realistic simulations of challenging detection tasks as well as clinical data of patients with intractable epilepsy. The outlier detection framework was further extended to take into account the specificities of neuroimaging data and the detection task at hand. We first proposed a reformulation of the support vector data description (SVDD) method to deal with the presence of uncertain observations in the training data. Second, to handle the multi-parametric nature of neuroimaging data, we proposed an optimal fusion approach for combining multiple base one-class classifiers. Finally, to help with score interpretation, threshold selection and score combination, we proposed to transform the score outputs of the outlier detection algorithm into well calibrated probabilities.

A presença que tem hoje a Medicina Nuclear como modalidade de obtenção de imagens médicas é muito importante e um dos principais procedimentos utilizados hoje nos centros de saúde, tendo como grande vantagem a capacidade de conseguir analisar o comportamento metabólico do paciente, fazendo possíveis diagnósticos precoces. Este projeto está baseado em imagens médicas obtidas através da modalidade PET (Positron Emission Tomography) a qual está tendo uma crescente difusão e aceitação. Para isso, temos desenvolvido uma estrutura integral de processamento de imagens tridimensionais PET, a qual está constituída por etapas consecutivas que se iniciam na obtenção das imagens padrões (gold standard), sendo utilizados volumes simulados ou phantoms do Ventrículo Esquerdo do Coração criadas como parte do projeto, assim como geradas a partir do software NCAT-4D. A seguir, nos volumes simulados, é introduzido ruído quântico tipo Poisson que é o ruído característico das imagens PET e feita uma análise que busca certificar que o ruído utilizado corresponde efetivamente ao ruído Poisson. Em sequência é executada a parte de pré-processamento, utilizando para este fim, um conjunto de filtros tais como o filtro da mediana, o filtro da Gaussiana ponderada e o filtro que mistura os conceitos da Transformada de Anscombe e o filtro pontual de Wiener. Posteriormente é aplicada a etapa de segmentação que é considerada a parte central da sequência de processamento. O processo de segmentação é baseado na teoria de Conectividade Fuzzy e para isso temos implementado quatro diferentes abordagens: Algoritmo Genérico, Algoritmo LIFO, Algoritmo kTetaFOEMS e o Algoritmo utilizando Pesos Dinâmicos. Sendo que os três primeiros algoritmos utilizam pesos específicos selecionados pelo usuário, foi preciso efetuar uma análise para determinar os melhores pesos de segmentação que se reflitam numa segmentação mais eficiente. Finalmente, para terminar a estrutura de processamento, um procedimento de avaliação foi utilizado como métrica para obter quantitativamente três parâmetros (Verdadeiro Positivo, Falso Positivo e Máxima Distância) que permitiram conhecer o nível de eficiência e precisão de nosso processo e do projeto em geral. Constatamos que os algoritmos implementados (filtros e algoritmos de segmentação) são bastante robustos e atingem ótimos resultados chegando-se a obter, para o caso do volume do Ventrículo Esquerdo simulado, taxas de VP e FP na ordem de 98.49 ± 0.27% e 2,19 ± 0.19%, respectivamente. Com o conjunto de procedimentos e escolhas feitas ao longo da estrutura de processamento, encerramos o projeto com a análise de um grupo de volumes produto de um exame PET real, obtendo a quantificação destes volumes. / The usefulness of Nuclear medicine nowadays as a modality to obtain medical images is very important, and it has turned into one of the main procedures utilized in Health Care Centers. Its great advantage is to analyze the metabolic behavior of the patient, by allowing early diagnosis. This project is based on medical images obtained by the PET modality (Positron Emission Tomography), which has won wide acceptance. Thus, we have developed an integral framework for processing Nuclear Medicine three-dimensional images of the PET modality, which is composed of consecutive steps that start with the generation of standard images (gold standard) by using simulated images or phantoms of the Left Ventricular Heart that were generated in this project, such as the ones obtained from the NCAT-4D software. Then Poisson quantum noise is introduced into the whole volume to simulate the characteristic noises in PET images and an analysis is performed in order to certify that the utilized noise is the Poisson noise effectively. Subsequently, the pre-processing is executed by using specific filters, such as the median filter, the weighted Gaussian filter, and the filter that joins the concepts of Anscombe Transformation and the Wiener filter. Then the segmentation, which is considered the most important and central part of the whole process, is implemented. The segmentation process is based on the Fuzzy Connectedness theory and for that purpose four different approaches were implemented: Generic algorithm, LIFO algorithm, kTetaFOEMS algorithm, and Dynamic Weight algorithm. Since the first three algorithms used specific weights that were selected by the user, an extra analysis was performed to determine the best segmentation constants that would reflect an accurate segmentation. Finally, at the end of the processing structure, an assessment procedure was used as a measurement tool to quantify some parameters that determined the level of efficiency and precision of our process and project. We have verified that the implemented algorithms (filters and segmentation algorithms) are fairly robust and achieve optimal results, assist to obtain, in the case of the Left Ventricular simulated, TP and FP rates in the order of 98.49 ± 0.27% and 2.19 ± 0.19%, respectively. With the set of procedures and choices made along of the processing structure, the project was concluded with the analysis of a volumes group from a real PET exam, obtaining the quantification of the volumes.

A presença da Medicina Nuclear como modalidade de obtenção de imagens médicas é um dos principais procedimentos utilizados hoje nos centros de saúde, tendo como grande vantagem a capacidade de analisar o comportamento metabólico do paciente, traduzindo-se em diagnósticos precoces. Entretanto, sabe-se que a quantificação em Medicina Nuclear é dificultada por diversos fatores, entre os quais estão a correção de atenuação, espalhamento, algoritmos de reconstrução e modelos assumidos. Neste contexto, o principal objetivo deste projeto foi melhorar a acurácia e a precisão na análise de imagens de PET/CT via processos realísticos e bem controlados. Para esse fim, foi proposta a elaboração de uma estrutura modular, a qual está composta por um conjunto de passos consecutivamente interligados começando com a simulação de phantoms antropomórficos 3D para posteriormente gerar as projeções realísticas PET/CT usando a plataforma GATE (com simulação de Monte Carlo), em seguida é aplicada uma etapa de reconstrução de imagens 3D, na sequência as imagens são filtradas (por meio do filtro de Anscombe/Wiener para a redução de ruído Poisson caraterístico deste tipo de imagens) e, segmentadas (baseados na teoria Fuzzy Connectedness). Uma vez definida a região de interesse (ROI) foram produzidas as Curvas de Atividade de Entrada e Resultante requeridas no processo de análise da dinâmica de compartimentos com o qual foi obtida a quantificação do metabolismo do órgão ou estrutura de estudo. Finalmente, de uma maneira semelhante imagens PET/CT reais fornecidas pelo Instituto do Coração (InCor) do Hospital das Clínicas da Faculdade de Medicina da Universidade de São Paulo (HC-FMUSP) foram analisadas. Portanto, concluiu-se que a etapa de filtragem tridimensional usando o filtro Anscombe/Wiener foi relevante e de alto impacto no processo de quantificação metabólica e em outras etapas importantes do projeto em geral. / The presence of Nuclear Medicine as a medical imaging modality is one of the main procedures utilized nowadays in medical centers, and the great advantage of that procedure is its capacity to analyze the metabolic behavior of the patient, resulting in early diagnoses. However, the quantification in Nuclear Medicine is known to be complicated by many factors, such as degradations due to attenuation, scattering, reconstruction algorithms and assumed models. In this context, the goal of this project is to improve the accuracy and the precision of quantification in PET/CT images by means of realistic and well-controlled processes. For this purpose, we proposed to develop a framework, which consists in a set of consecutively interlinked steps that is initiated with the simulation of 3D anthropomorphic phantoms. These phantoms were used to generate realistic PET/CT projections by applying the GATE platform (with Monte Carlo simulation). Then a 3D image reconstruction was executed, followed by a filtering process (using the Anscombe/Wiener filter to reduce Poisson noise characteristic of this type of images) and, a segmentation process (based on the Fuzzy Connectedness theory). After defining the region of interest (ROI), input activity and output response curves are required for the compartment analysis in order to obtain the Metabolic Quantification of the selected organ or structure. Finally, in the same manner real images provided from the Heart Institute (InCor) of Hospital das Clínicas, Faculty of Medicine, University of São Paulo (HC-FMUSP) were analysed. Therefore, it is concluded that the three-dimensional filtering step using the Ascombe/Wiener filter was preponderant and had a high impact on the metabolic quantification process and on other important stages of the whole project.

Positron emission tomography (PET) is a functional imaging method that allows studying physiological, biochemical or pharmacological processes in vivo. PET is being used in both research and clinical practice. In the brain, it has been used to investigate metabolism, receptor binding, and alterations in regional blood flow. This thesis involves both preclinical and clinical dual-tracer PET imaging studies of different neurological disorders. In this way, different radiotracers were used along the projects. The first project focused on the implementation and in vivo validation of the simultaneous dual-tracer PET imaging technique on the rat brain and its applications in the study of cerebral ischemia. In particular, in this project two biological processes were studied at the same time: cerebral blood flow and cerebral glucose metabolism. The second project consisted in a clinical correlation study of the GABAergic and serotonin systems in a population with Essential Tremor (ET), the most commonly movement disorders. / La tomografia per emissió de positrons (PET) és un mètode d'imatge funcional que permet l'estudi in vivo de processos fisiològics, bioquímics i farmacològics. La PET s'utilitza tant en la pràctica clínica com en la recerca. Al cervell, s'ha utilitzat per investigar el metabolisme, la neurotransmissió, i les alteracions en el flux sanguini regional. Aquesta tesi implica estudis preclínics i clínics de la tècnica PET en diversos trastorns neurològics. D'aquesta manera, es van utilitzar diferents radiotraçadors al llarg dels projectes. El primer projecte es va centrar en la implementació i validació in vivo de la tècnica PET del doble-marcador simultani en el cervell de rata i les seves aplicacions en l'estudi de la isquèmia cerebral. En particular, en aquest projecte es van estudiar en el mateix moment dos processos biològics: el flux sanguini cerebral i el metabolisme cerebral de la glucosa. El segon projecte va consistir en un estudi clínic de correlació dels sistemes GABAèrgic i serotoninèrgic en una població amb tremolor essencial (TE), el trastorn del moviment més comú

Positron emission tomography (PET) is a well-established functional imaging method used in nuclear medicine. It allows for retrieving information about biochemical and physiological processes in vivo. The currently possible spatial resolution of PET is about 5 mm for brain acquisitions and about 8 mm for whole-body acquisitions, while recent improvements in image reconstruction point to a resolution of 2 mm in the near future. Typical acquisition times range from minutes to hours due to the low signal-to-noise ratio of the measuring principle, as well as due to the monitoring of the metabolism of the patient over a certain time. Therefore, patient motion increasingly limits the possible spatial resolution of PET. In addition, patient immobilisations are only of limited benefit in this context. Thus, patient motion leads to a relevant resolution degradation and incorrect quantification of metabolic parameters. The present work describes the utilisation of a novel motion compensation method for clinical brain PET acquisitions. By using an external motion tracking system, information about the head motion of a patient is continuously acquired during a PET acquisition. Based on the motion information, a newly developed event-based motion compensation algorithm performs spatial transformations of all registered coincidence events, thus utilising the raw data of a PET system - the so-called `list-mode´ data. For routine acquisition of this raw data, methods have been developed which allow for the first time to acquire list-mode data from an ECAT Exact HR+ PET scanner within an acceptable time frame. Furthermore, methods for acquiring the patient motion in clinical routine and methods for an automatic analysis of the registered motion have been developed. For the clinical integration of the aforementioned motion compensation approach, the development of additional methods (e.g. graphical user interfaces) was also part of this work. After development, optimisation and integration of the event-based motion compensation in clinical use, analyses with example data sets have been performed. Noticeable changes could be demonstrated by analysis of the qualitative and quantitative effects after the motion compensation. From a qualitative point of view, image artefacts have been eliminated, while quantitatively, the results of a tracer kinetics analysis of a FDOPA acquisition showed relevant changes in the R0k3 rates of an irreversible reference tissue two compartment model. Thus, it could be shown that an integration of a motion compensation method which is based on the utilisation of the raw data of a PET scanner, as well as the use of an external motion tracking system, is not only reasonable and possible for clinical use, but also shows relevant qualitative and quantitative improvement in PET imaging. / Die Positronen-Emissions-Tomographie (PET) ist ein in der Nuklearmedizin etabliertes funktionelles Schnittbildverfahren, das es erlaubt Informationen über biochemische und physiologische Prozesse in vivo zu erhalten. Die derzeit erreichbare räumliche Auflösung des Verfahrens beträgt etwa 5 mm für Hirnaufnahmen und etwa 8 mm für Ganzkörperaufnahmen, wobei erste verbesserte Bildrekonstruktionsverfahren eine Machbarkeit von 2 mm Auflösung in Zukunft möglich erscheinen lassen. Durch das geringe Signal/Rausch-Verhältnis des Messverfahrens, aber auch durch die Tatsache, dass der Stoffwechsel des Patienten über einen längeren Zeitraum betrachtet wird, betragen typische PET-Aufnahmezeiten mehrere Minuten bis Stunden. Dies hat zur Folge, dass Patientenbewegungen zunehmend die erreichbare räumliche Auflösung dieses Schnittbildverfahrens limitieren. Eine Immobilisierung des Patienten zur Reduzierung dieser Effekte ist hierbei nur bedingt hilfreich. Es kommt daher zu einer relevanten Auflösungsverschlechterung sowie zu einer Verfälschung der quantifizierten Stoffwechselparameter. Die vorliegende Arbeit beschreibt die Nutzbarmachung eines neuartigen Bewegungskorrekturverfahrens für klinische PET-Hirnaufnahmen. Mittels eines externen Bewegungsverfolgungssystems wird während einer PET-Untersuchung kontinuierlich die Kopfbewegung des Patienten registriert. Anhand dieser Bewegungsdaten führt ein neu entwickelter event-basierter Bewegungskorrekturalgorithmus eine räumliche Korrektur aller registrierten Koinzidenzereignisse aus und nutzt somit die als "List-Mode" bekannten Rohdaten eines PET Systems. Für die Akquisition dieser Daten wurden eigens Methoden entwickelt, die es erstmals erlauben, diese Rohdaten von einem ECAT Exact HR+ PET Scanner innerhalb eines akzeptablen Zeitraumes zu erhalten. Des Weiteren wurden Methoden für die klinische Akquisition der Bewegungsdaten sowie für die automatische Auswertung dieser Daten entwickelt. Ebenfalls Teil der Arbeit waren die Entwicklung von Methoden zur Integration in die klinische Routine (z.B. graphische Nutzeroberflächen). Nach der Entwicklung, Optimierung und Integration der event-basierten Bewegungskorrektur für die klinische Nutzung wurden Analysen anhand von Beispieldatensätzen vorgenommen. Es zeigten sich bei der Auswertung sowohl der qualitativen als auch der quantitativen Effekte deutliche Änderungen. In qualitativer Hinsicht wurden Bildartefakte eliminiert; bei der quantitativen Auswertung einer FDOPA Messung zeigte sich eine revelante Änderung der R0k3 Einstromraten eines irreversiblen Zweikompartment-Modells mit Referenzgewebe. Es konnte somit gezeigt werden, dass eine Integration einer Bewegungskorrektur unter Zuhilfenahme der Rohdaten eines PET Systems sowie unter Nutzung eines externen Verfolgungssystems nicht nur sinnvoll und in der klinischen Routine machbar ist, sondern auch zu maßgeblichen qualitativen und quantitativen Verbesserungen in der PET-Bildgebung beitragen kann.

[EN] Positron Emission Tomography (PET) is a non-invasive nuclear medical imaging modality that makes it possible to observe the distribution of metabolic substances within a patient's body after marking them with radioactive isotopes and arranging an annular scanner around him in order to detect their decays. The main applications of this technique are the detection and tracing of tumors in cancer patients and metabolic studies with small animals. The Electronic Design for Nuclear Applications (EDNA) research group within the Instituto de Instrumentación para Imagen Molecular (I3M) has been involved in the study of high performance PET systems and maintains a small experimental setup with two detector modules. This thesis is framed within the necessity of developing a new data acquisition system (DAQ) for the aforementioned setup that corrects the drawbacks of the existing one. The main objective is to define a DAQ architecture that is completely scalable, modular, and guarantees the mobility and the possibility of reusing its components, so that it admits any extension of modification of the setup and it is possible to export it directly to the configurations used by other groups or experiments. At the same time, this architecture should be compatible with the best possible resolutions attainable at the present instead of imposing artificial limits on system performance. In particular, the new DAQ system should outperform the previous one. As a first step, a general study of DAQ arquitectures is carried out in the context of experimental setups for PET and other high energy physics applications. On one hand, the conclusion is reached that the desired specifications require early digitization of detector signals, exclusively digital communication between modules, and the absence of a centralized trigger. On the other hand, the necessity of a very precise distributed synchronization scheme between modules becomes apparent, with errors in the order of 100 ps, and operating directly over the data links. A study of the existing methods reveals their severe limitations in terms of achievable precision. A theoretical analysis of the situation is carried out with the goal of overcoming them, and a new synchronization algorithm is proposed that is able to reach the desired resolution while getting rid of the restrictions on clock alignment that are imposed by virtually all usual schemes. Since the measurement of clock phase difference plays a crucial role in the proposed algorithm, extensions to the existing methods are defined and analyzed that improve them significantly. The proposed scheme for synchronism is validated using commercial evaluation boards. Taking the proposed synchronization method as a starting point, a DAQ architecture for PET is defined that is composed of two types of module (acquisition and concentration) whose replication makes it possible to arrange a hierarchic system of arbitrary size, and circuit boards are designed and commissioned that implement a realization of the architecture for the particular case of two detectors. This DAQ is finally installed at the experimental setup, where their synchronization properties and resolution as a PET system are characterized and its performance is verified to have improved with respect to the previous system. / [ES] La Tomografía por Emisión de Positrones (PET) es una modalidad de imagen médica nuclear no invasiva que permite observar la distribución de sustancias metabólicas en el interior del cuerpo de un paciente tras marcarlas con isótopos radioactivos y disponer después un escáner anular a su alrededor para detectar su desintegración. Las principales aplicaciones de esta técnica son la detección y seguimiento de tumores en pacientes con cáncer y los estudios metabólicos en animales pequeños. El grupo de investigación Electronic Design for Nuclear Applications (EDNA) del Instituto de Instrumentación para Imagen Molecular (I3M) ha estado involucrado en el estudio de sistemas PET de alto rendimiento y mantiene un pequeño setup experimental con dos módulos detectores. La presente tesis se enmarca dentro de la necesidad de desarrollar un nuevo sistema de adquisición de datos (DAQ) para dicho setup que corrija los inconvenientes del ya existente. En particular, el objetivo es definir una arquitectura de DAQ que sea totalmente escalable, modular, y que asegure la movilidad y la posibilidad de reutilización de sus componentes, de manera que admita cualquier ampliación o alteración del setup y pueda exportarse directamente a los de otros grupos o experimentos. Al mismo tiempo, se desea que dicha arquitectura no limite artificialmente el rendimiento del sistema sino que sea compatible con las mejores resoluciones disponibles en la actualidad, y en particular que sus prestaciones superen a las del DAQ instalado previamente. En primer lugar, se lleva a cabo un estudio general de las arquitecturas de DAQ para setups experimentales para PET y otras aplicaciones de física de altas energías. Por un lado, se determina que las características deseadas implican la digitalización temprana de las señales del detector, la comunicación exclusivamente digital entre módulos, y la ausencia de trigger centralizado. Por otro lado, se hace patente la necesidad de un esquema de sincronización distribuida muy preciso entre módulos, con errores del orden de 100 ps, que opere directamente sobre los enlaces de datos. Un estudio de los métodos ya existentes revela sus graves limitaciones a la hora de alcanzar esas precisiones. Con el fin de paliarlos, se lleva a cabo un análisis teórico de la situación y se propone un nuevo algoritmo de sincronización que es capaz de alcanzar la resolución deseada y elimina las restricciones de alineamiento de reloj impuestas por casi todos los esquemas usuales. Dado que la medida de desfase entre relojes juega un papel crucial en el algoritmo propuesto, se definen y analizan extensiones a los métodos ya existentes que suponen una mejora sustancial. El esquema de sincronismo propuesto se valida utilizando placas de evaluación comerciales. Partiendo del método de sincronismo propuesto, se define una arquitectura de DAQ para PET compuesta de dos tipos de módulos (adquisición y concentración) cuya replicación permite construir un sistema jerárquico de tamaño arbitrario, y se diseñan e implementan placas de circuito basadas en dicha arquitectura para el caso particular de dos detectores. El DAQ así construído se instala finalmente en el setup experimental, donde se caracterizan tanto sus propiedades de sincronización como su resolución como sistema PET y se comprueba que sus prestaciones son superiores a las del sistema previo. / [CA] La Tomografia per Emissió de Positrons (PET) és una modalitat d'imatge mèdica nuclear no invasiva que permet observar la distribució de substàncies metabòliques a l'interior del cos d'un pacient després d'haver-les marcat amb isòtops radioactius disposant un escàner anular al seu voltant per a detectar la seua desintegració. Aquesta tècnica troba les seues principals aplicacions a la detecció i seguiment de tumors a pacients amb càncer i als estudis metabòlics en animals petits. El grup d'investigació Electronic Design for Nuclear Applications (EDNA) de l'Instituto de Instrumentación para Imagen Molecular (I3M) ha estat involucrat en l'estudi de sistemes PET d'alt rendiment i manté un petit setup experimental amb dos mòduls detectors. Aquesta tesi neix de la necessitat de desenvolupar un nou sistema d'adquisició de dades (DAQ) per al setup esmentat que corregisca els inconvenients de l'anterior. En particular, l'objectiu és definir una arquitectura de DAQ que sigui totalment escalable, modular, i que asseguri la mobilitat i la possibilitat de reutilització dels seus components, de tal manera que admeta qualsevol ampliació o alteració del setup i pugui exportar-se directament a aquells d'altres grups o experiments. Al mateix temps, es desitja que aquesta arquitectura no introduisca límits artificials al rendiment del sistema sinó que sigui compatible amb les millors resolucions disponibles a l'actualitat, i en particular que les seues prestacions siguin superiors a les del DAQ instal.lat amb anterioritat. En primer lloc, es porta a terme un estudi general de les arquitectures de DAQ per a setups experimentals per a PET i altres aplicacions de física d'altes energies. Per una banda, s'arriba a la conclusió que les característiques desitjades impliquen la digitalització dels senyals del detector el més aviat possible, la comunicació exclusivament digital entre mòduls, i l'absència de trigger centralitzat. D'altra banda, es fa palesa la necessitat d'un mecanisme de sincronització distribuïda molt precís entre mòduls, amb errors de l'ordre de 100 ps, que treballi directament sobre els enllaços de dades. Un estudi dels mètodes ja existents revela les seues greus limitacions a l'hora d'assolir aquest nivell de precisió. Amb l'objectiu de pal.liar-les, es duu a terme una anàlisi teòrica de la situació i es proposa un nou algoritme de sincronització que és capaç d'obtindre la resolució desitjada i es desfà de les restriccions d'alineament de rellotges imposades per gairebé tots els esquemes usuals. Atès que la mesura del desfasament entre rellotges juga un paper cabdal a l'algoritme proposat, es defineixen i analitzen extensions als mètodes ja existents que suposen una millora substancial. L'esquema de sincronisme proposat es valida mitjançant plaques d'avaluació comercials. Prenent el mètode proposat com a punt de partida, es defineix una arquitectura de DAQ per a PET composta de dos tipus de mòduls (d'adquisició i de concentració) tals que la replicació d'aquests elements permet construir un sistema jeràrquic de mida arbitrària, i es dissenyen i implementen plaques de circuit basades en aquesta arquitectura per al cas particular de dos detectors. L'electrònica desenvolupada s'instal.la finalment al setup experimental, on es caracteritzen tant les seues propietats de sincronització com la seua resolució com a sistema PET i es comprova que les seues prestacions són superiors a les del sistema previ. / Aliaga Varea, RJ. (2016). Development of a data acquisition architecture with distributed synchronization for a Positron Emission Tomography system with integrated front-end [Tesis doctoral]. Universitat Politècnica de València. https://doi.org/10.4995/Thesis/10251/63271 / Premios Extraordinarios de tesis doctorales

Die Interaktion von S100-Proteinen mit dem Rezeptor für advanced glycation endproducts (RAGE) wird als hoch relevant bei der Entstehung, Manifestation und Progression verschiedener entzündlicher Erkrankungen sowie bei der Tumorigenese gewertet. Das tiefergehende Verständnis der Interaktion von S100-Proteinen mit RAGE in vivo stellt eine wissenschaftliche Herausforderung dar und ist ein Ansatz für therapeutische Interventionen. Darüber hinaus stellen Untersuchungen zum Metabolismus von extrazellulär zirkulierenden S100-Proteinen in vivo einen vielversprechenden Forschungsansatz zur Analyse von S100-Protein-assoziierten Erkrankungen dar. Die einzigartigen Eigenschaften der Positronen-Emissions-Tomographie (PET) als nicht-invasives bildgebendes Verfahren erlauben die Darstellung und quantitative Erfassung biochemischer Prozesse mit der Möglichkeit zelluläre und molekulare Reaktionswege aufzuzeigen sowie in vivo-Mechanismen von Krankheiten im Kontext eines physiologischen Umfeldes darzulegen. Ziel der vorliegenden Arbeit war es, Fluor-18-markierte S100-Proteine (18F-S100) herzustellen, diese biochemisch, radiochemisch und radiopharmakologisch zu charakterisieren und deren Metabolismus und Interaktion mit RAGE in vivo mittels Kleintier-PET am Tiermodell zu untersuchen. Es wurden die mit RAGE interagierenden S100-Proteine S100A1, S100A12 und S100B in biologisch funktioneller Form hergestellt. Dazu wurden die entsprechenden S100-Gene in den prokaryotischen Expressionsvektor pGEX-6P-1 kloniert. Mit diesen Konstrukten wurden E. coli-Zellen transformiert, aus denen nachfolgend die S100-Proteine isoliert und gereinigt werden konnten. Es konnte eine Reinigung unter nativen, milden Bedingungen etabliert werden, die es ermöglichte, S100A1, S100A12 und S100B in biologisch aktiver Form und in hohen Reinheitsgraden (> 95%) für die nachfolgenden Experimente bereitzustellen. Diese S100-Proteine wurden über den 18F-tragenden Aktivester N-Succinimidyl-4-[18F]fluorbenzoesäure ([18F]SFB) radioaktiv markiert und charakterisiert. Dabei konnte sichergestellt werden, dass die 18F-S100-Proteine in vitro und in vivo stabil sind. Weiterhin konnte nachgewiesen werden, dass die radioaktive Markierung keine Beeinträchtigung auf die biologische Funktionalität der S100-Proteine hat. Dies wurde anhand von sRAGE-Bindungsuntersuchungen sowie Zell-Interaktionsuntersuchungen an konfluenten Endothelzellen (HAEC) und an zu Makrophagen differenzierten THP-1-Zellen (THP-1-Makrophagen) verifiziert. Für die Untersuchung der RAGE-Bindung war die Produktion des löslichen sRAGE bzw. die Generation von flRAGE-berexprimierenden Zellen erforderlich. Beide Konstrukte wurden in geeigneten Zellsystemen exprimiert und das sRAGE-Protein wurde in biologisch aktiver Form synthetisiert und gereinigt (Reinheitsgrad > 97%). Die 18F-S100-Bindung an THP-1-Makrophagen und HAEC wurde in Gegenwart von glykierten LDL (glykLDL) sowie sRAGE signifikant inhibiert, was auf eine RAGE-Interaktion hinweist. Weiterhin konnten durch den Einsatz von Scavenger-Rezeptor-Liganden, wie z. B. Maleinanhydrid-modifiziertes BSA (malBSA) bzw. von Lektinen inhibierende Effekte erzielt werden. Dies ist ein Indiz für die 18F-S100-Interaktion mit Scavenger-Rezeptoren und Glykokonjugaten an der Zelloberfläche. Durch die Untersuchungen mittels konfokaler Laserscanning-Mikroskopie an THP-1-Makrophagen wurde eine Zellaufnahme des Fluoreszein-markierten S100A12 festgestellt. Weiterhin konnten Kolokalisationen mit Lektinen detektiert werden. Das metabolische Schicksal extrazellulär zirkulierender 18F-S100-Proteine in vivo wurde mit Hilfe dynamischer PET-Untersuchungen bzw. anhand von Bioverteilungs-Untersuchungen in männlichen Wistar-Ratten analysiert. Die Hauptakkumulation der Radioaktivität wurde in der Leber und in den Nieren detektiert. In diesen Organen findet der Metabolismus bzw. die glomeruläre Filtration der 18F-S100-Proteine statt. In den Untersuchungen zur Genexpression mittels Echtzeit-PCR sowie im immunchemischen Proteinnachweis am Western Blot wurde eine hohe Expression und Proteinbiosynthese des RAGE in der Lunge ermittelt. Die Lunge eignet sich daher als „Referenz“-Organ für eine funktionelle in vivo-Charakterisierung von RAGE mit 18FS100-Proteinen. Bei den durchgeführten PET-Untersuchungen konnte eine temporäre 18F-S100-Interaktion mit dem Lungengewebe festgestellt werden. Die Retention des 18FS100A12 in der Lunge wurde in Gegenwart von sRAGE inhibiert. Dies ist ein Hinweis dafür, dass 18F-S100-Proteine auch in vivo an RAGE binden können. Die Radioaktivitäts-Akkumulation in den Organen Leber und Milz, die eine Vielzahl von sessilen Makrophagen aufweisen, wurde durch die Applikation von malBSA inhibiert. Dies ist ein Indiz dafür, dass 18F-S100-Proteine in vivo mit Scavenger-Rezeptoren interagieren können. Die vorliegende Arbeit liefert deutliche Hinweise darauf, dass RAGE nicht der alleinige Rezeptor für 18F-S100-Proteine ist. Der Einsatz von 18F-S100-Proteinen als experimentelles Werkzeug in dynamischen PET-Untersuchungen birgt das Potential einer Charakterisierung von S100-Protein-assoziierten, pathophysiologischen Prozessen. / Members of the S100 family of EF-hand calcium binding proteins play important regulatory roles not only within cells but also exert effects in a cytokine-like manner on definite target cells once released into extracellular space or circulating blood. Accordingly, increased levels of S100 proteins in the circulating blood have been associated with a number of disease states, e.g., diabetes, cancer, and various inflammatory disorders. As the best known target protein of extracellular S100 proteins, the receptor for advanced glycation endproducts (RAGE) is of significant importance. However, the role of extracellular S100 proteins during etiology, progression, and manifestation of inflammatory disorders still is poorly understood. One reason for this is the shortage of sensitive methods for direct assessment of the metabolic fate of circulating S100 proteins and, on the other hand, measurement of functional expression of extracellular targets of S100 proteins, e.g., RAGE in vivo. In this line, small animal PET provides a valuable tool for noninvasive imaging of physiological processes and interactions like plasma or vascular retention, tissue-specific receptor binding, accumulation or elimination in vivo. To address this question, human S100 proteins were cloned in the bacterial expression vector pGEX-6P-1, expressed in E. coli BL21, and purified by affinity chromatography and anion exchange chromatography. Purified S100A1, S100B and S100A12 proteins were then radiolabeled with the positron emitter fluorine-18 (18F) by N-succinimidyl-4-[18F]fluorobenzoate ([18F]SFB). Radiolabeling of S100 proteins resulted in radiochemical yields of 3-10% (corrected for decay) and effective specific radioactivities of 1 GBq/µmol, respectively. For investigations about RAGE binding soluble RAGE (sRAGE) was expressed and purified using pSecTag2B. A radioligand binding assay confirmed specific binding of 18F-S100A12, 18F-S100A1, and 18F-S100B to immobilized sRAGE, also showing an order of affinity with S100A12 > S100A1 > S100B. These results indicate that radioactive labelling of S100 proteins did not affect their overall affinity to RAGE. Cellular association studies in human THP-1 macrophages and human aortic endothelial cells (HAEC) showed specific binding of all 18F-S100 proteins to the non-internalizing RAGE as confirmed by inhibitory effects exerted either by other RAGE ligands, e.g., glycated LDL, or by soluble RAGE. Of interest, 18F-S100 proteins were also shown to interact with other putative binding sites, e.g. scavenger receptors as well as proteoglycans. In this line, uptake of 18F-S100 proteins in THP-1 and HAEC could be inhibited by various scavenger receptor ligands, in particular by maleylated BSA as well as by lectines (e.g. ConA and SBA). Confocal laser scanning microscopy analysis showed a major part of the fluoresceinated S100A12 bound to the surface of THP-1 macrophages. Beyond this, uptake of S100A12 could be determined indicating an interaction of S100A12 with both non-internalizing, e.g., RAGE, and internalizing receptors, e.g. scavenger receptors. By evaluation of the relative contribution of 18F-S100A12 association to RAGE-overexpressed CHO cells (using pIres2-AcGFP1), 18F-S100A12 showed a significantly higher association to CHO-RAGE cells compared with CHO-mock cells. Based on these findings and due to their crucial role in inflammatory disorders the metabolic fate of S100 proteins was further investigated in dynamic small animal Positron emission tomography (PET) studies as well as in biodistribution studies in Wistar rats in vivo. For interpretation of in vivo investigations in rats, expression of RAGE was analyzed by quantitative real time RT-PCR as well as western blotting in various organs. Lung tissue expressed the highest level of RAGE protein compared to the other tissues. PET studies in rats revealed a comparatively long mean residence time of circulating 18F-S100 proteins. A major contributor to this phenomenon seems to be a sustained temporary interaction with tissues overexpressing RAGE, e.g., the lung. On the other hand, renal clearance of 18F-S100 via glomerular filtration is a major elimination pathway. However, scavenger receptor-mediated pathways in the liver, the spleen and, to a minor extent, in the kidneys, also seem to contribute to the overall clearance. The presence of sRAGE revealed a decreased retention of 18F-S100A12 in the lung, indicating in vivo binding to RAGE. In vivo blocking studies using maleylated BSA demonstrated a strong inhibition of putative binding sites in rat tissues enriched in cells expressing scavenger receptors like liver and spleen. In conclusion, 18F-labeling of S100 proteins and the use of small animal PET provide a valuable tool to discriminate the kinetics and the metabolic fate of S100 proteins in vivo. Furthermore, the results strongly suggest an involvement of other putative receptors beside RAGE in distribution, tissue association and elimination of circulating proinflammatory S100 proteins. Moreover, the approach provides novel probes for imaging of functional expression of RAGE and scavenger receptors in peripheral inflammatory compartments.

Die Interaktion von S100-Proteinen mit dem Rezeptor für advanced glycation endproducts (RAGE) wird als hoch relevant bei der Entstehung, Manifestation und Progression verschiedener entzündlicher Erkrankungen sowie bei der Tumorigenese gewertet. Das tiefergehende Verständnis der Interaktion von S100-Proteinen mit RAGE in vivo stellt eine wissenschaftliche Herausforderung dar und ist ein Ansatz für therapeutische Interventionen. Darüber hinaus stellen Untersuchungen zum Metabolismus von extrazellulär zirkulierenden S100-Proteinen in vivo einen vielversprechenden Forschungsansatz zur Analyse von S100-Protein-assoziierten Erkrankungen dar. Die einzigartigen Eigenschaften der Positronen-Emissions-Tomographie (PET) als nicht-invasives bildgebendes Verfahren erlauben die Darstellung und quantitative Erfassung biochemischer Prozesse mit der Möglichkeit zelluläre und molekulare Reaktionswege aufzuzeigen sowie in vivo-Mechanismen von Krankheiten im Kontext eines physiologischen Umfeldes darzulegen. Ziel der vorliegenden Arbeit war es, Fluor-18-markierte S100-Proteine (18F-S100) herzustellen, diese biochemisch, radiochemisch und radiopharmakologisch zu charakterisieren und deren Metabolismus und Interaktion mit RAGE in vivo mittels Kleintier-PET am Tiermodell zu untersuchen. Es wurden die mit RAGE interagierenden S100-Proteine S100A1, S100A12 und S100B in biologisch funktioneller Form hergestellt. Dazu wurden die entsprechenden S100-Gene in den prokaryotischen Expressionsvektor pGEX-6P-1 kloniert. Mit diesen Konstrukten wurden E. coli-Zellen transformiert, aus denen nachfolgend die S100-Proteine isoliert und gereinigt werden konnten. Es konnte eine Reinigung unter nativen, milden Bedingungen etabliert werden, die es ermöglichte, S100A1, S100A12 und S100B in biologisch aktiver Form und in hohen Reinheitsgraden (> 95%) für die nachfolgenden Experimente bereitzustellen. Diese S100-Proteine wurden über den 18F-tragenden Aktivester N-Succinimidyl-4-[18F]fluorbenzoesäure ([18F]SFB) radioaktiv markiert und charakterisiert. Dabei konnte sichergestellt werden, dass die 18F-S100-Proteine in vitro und in vivo stabil sind. Weiterhin konnte nachgewiesen werden, dass die radioaktive Markierung keine Beeinträchtigung auf die biologische Funktionalität der S100-Proteine hat. Dies wurde anhand von sRAGE-Bindungsuntersuchungen sowie Zell-Interaktionsuntersuchungen an konfluenten Endothelzellen (HAEC) und an zu Makrophagen differenzierten THP-1-Zellen (THP-1-Makrophagen) verifiziert. Für die Untersuchung der RAGE-Bindung war die Produktion des löslichen sRAGE bzw. die Generation von flRAGE-berexprimierenden Zellen erforderlich. Beide Konstrukte wurden in geeigneten Zellsystemen exprimiert und das sRAGE-Protein wurde in biologisch aktiver Form synthetisiert und gereinigt (Reinheitsgrad > 97%). Die 18F-S100-Bindung an THP-1-Makrophagen und HAEC wurde in Gegenwart von glykierten LDL (glykLDL) sowie sRAGE signifikant inhibiert, was auf eine RAGE-Interaktion hinweist. Weiterhin konnten durch den Einsatz von Scavenger-Rezeptor-Liganden, wie z. B. Maleinanhydrid-modifiziertes BSA (malBSA) bzw. von Lektinen inhibierende Effekte erzielt werden. Dies ist ein Indiz für die 18F-S100-Interaktion mit Scavenger-Rezeptoren und Glykokonjugaten an der Zelloberfläche. Durch die Untersuchungen mittels konfokaler Laserscanning-Mikroskopie an THP-1-Makrophagen wurde eine Zellaufnahme des Fluoreszein-markierten S100A12 festgestellt. Weiterhin konnten Kolokalisationen mit Lektinen detektiert werden. Das metabolische Schicksal extrazellulär zirkulierender 18F-S100-Proteine in vivo wurde mit Hilfe dynamischer PET-Untersuchungen bzw. anhand von Bioverteilungs-Untersuchungen in männlichen Wistar-Ratten analysiert. Die Hauptakkumulation der Radioaktivität wurde in der Leber und in den Nieren detektiert. In diesen Organen findet der Metabolismus bzw. die glomeruläre Filtration der 18F-S100-Proteine statt. In den Untersuchungen zur Genexpression mittels Echtzeit-PCR sowie im immunchemischen Proteinnachweis am Western Blot wurde eine hohe Expression und Proteinbiosynthese des RAGE in der Lunge ermittelt. Die Lunge eignet sich daher als „Referenz“-Organ für eine funktionelle in vivo-Charakterisierung von RAGE mit 18FS100-Proteinen. Bei den durchgeführten PET-Untersuchungen konnte eine temporäre 18F-S100-Interaktion mit dem Lungengewebe festgestellt werden. Die Retention des 18FS100A12 in der Lunge wurde in Gegenwart von sRAGE inhibiert. Dies ist ein Hinweis dafür, dass 18F-S100-Proteine auch in vivo an RAGE binden können. Die Radioaktivitäts-Akkumulation in den Organen Leber und Milz, die eine Vielzahl von sessilen Makrophagen aufweisen, wurde durch die Applikation von malBSA inhibiert. Dies ist ein Indiz dafür, dass 18F-S100-Proteine in vivo mit Scavenger-Rezeptoren interagieren können. Die vorliegende Arbeit liefert deutliche Hinweise darauf, dass RAGE nicht der alleinige Rezeptor für 18F-S100-Proteine ist. Der Einsatz von 18F-S100-Proteinen als experimentelles Werkzeug in dynamischen PET-Untersuchungen birgt das Potential einer Charakterisierung von S100-Protein-assoziierten, pathophysiologischen Prozessen. / Members of the S100 family of EF-hand calcium binding proteins play important regulatory roles not only within cells but also exert effects in a cytokine-like manner on definite target cells once released into extracellular space or circulating blood. Accordingly, increased levels of S100 proteins in the circulating blood have been associated with a number of disease states, e.g., diabetes, cancer, and various inflammatory disorders. As the best known target protein of extracellular S100 proteins, the receptor for advanced glycation endproducts (RAGE) is of significant importance. However, the role of extracellular S100 proteins during etiology, progression, and manifestation of inflammatory disorders still is poorly understood. One reason for this is the shortage of sensitive methods for direct assessment of the metabolic fate of circulating S100 proteins and, on the other hand, measurement of functional expression of extracellular targets of S100 proteins, e.g., RAGE in vivo. In this line, small animal PET provides a valuable tool for noninvasive imaging of physiological processes and interactions like plasma or vascular retention, tissue-specific receptor binding, accumulation or elimination in vivo. To address this question, human S100 proteins were cloned in the bacterial expression vector pGEX-6P-1, expressed in E. coli BL21, and purified by affinity chromatography and anion exchange chromatography. Purified S100A1, S100B and S100A12 proteins were then radiolabeled with the positron emitter fluorine-18 (18F) by N-succinimidyl-4-[18F]fluorobenzoate ([18F]SFB). Radiolabeling of S100 proteins resulted in radiochemical yields of 3-10% (corrected for decay) and effective specific radioactivities of 1 GBq/µmol, respectively. For investigations about RAGE binding soluble RAGE (sRAGE) was expressed and purified using pSecTag2B. A radioligand binding assay confirmed specific binding of 18F-S100A12, 18F-S100A1, and 18F-S100B to immobilized sRAGE, also showing an order of affinity with S100A12 > S100A1 > S100B. These results indicate that radioactive labelling of S100 proteins did not affect their overall affinity to RAGE. Cellular association studies in human THP-1 macrophages and human aortic endothelial cells (HAEC) showed specific binding of all 18F-S100 proteins to the non-internalizing RAGE as confirmed by inhibitory effects exerted either by other RAGE ligands, e.g., glycated LDL, or by soluble RAGE. Of interest, 18F-S100 proteins were also shown to interact with other putative binding sites, e.g. scavenger receptors as well as proteoglycans. In this line, uptake of 18F-S100 proteins in THP-1 and HAEC could be inhibited by various scavenger receptor ligands, in particular by maleylated BSA as well as by lectines (e.g. ConA and SBA). Confocal laser scanning microscopy analysis showed a major part of the fluoresceinated S100A12 bound to the surface of THP-1 macrophages. Beyond this, uptake of S100A12 could be determined indicating an interaction of S100A12 with both non-internalizing, e.g., RAGE, and internalizing receptors, e.g. scavenger receptors. By evaluation of the relative contribution of 18F-S100A12 association to RAGE-overexpressed CHO cells (using pIres2-AcGFP1), 18F-S100A12 showed a significantly higher association to CHO-RAGE cells compared with CHO-mock cells. Based on these findings and due to their crucial role in inflammatory disorders the metabolic fate of S100 proteins was further investigated in dynamic small animal Positron emission tomography (PET) studies as well as in biodistribution studies in Wistar rats in vivo. For interpretation of in vivo investigations in rats, expression of RAGE was analyzed by quantitative real time RT-PCR as well as western blotting in various organs. Lung tissue expressed the highest level of RAGE protein compared to the other tissues. PET studies in rats revealed a comparatively long mean residence time of circulating 18F-S100 proteins. A major contributor to this phenomenon seems to be a sustained temporary interaction with tissues overexpressing RAGE, e.g., the lung. On the other hand, renal clearance of 18F-S100 via glomerular filtration is a major elimination pathway. However, scavenger receptor-mediated pathways in the liver, the spleen and, to a minor extent, in the kidneys, also seem to contribute to the overall clearance. The presence of sRAGE revealed a decreased retention of 18F-S100A12 in the lung, indicating in vivo binding to RAGE. In vivo blocking studies using maleylated BSA demonstrated a strong inhibition of putative binding sites in rat tissues enriched in cells expressing scavenger receptors like liver and spleen. In conclusion, 18F-labeling of S100 proteins and the use of small animal PET provide a valuable tool to discriminate the kinetics and the metabolic fate of S100 proteins in vivo. Furthermore, the results strongly suggest an involvement of other putative receptors beside RAGE in distribution, tissue association and elimination of circulating proinflammatory S100 proteins. Moreover, the approach provides novel probes for imaging of functional expression of RAGE and scavenger receptors in peripheral inflammatory compartments.