Integração e contextualização de modelos 3D no ensino da área da saúde

Mestrado em Engenharia Informática. Sistemas Gráficos e Multimédia.

Bioactive beads for local sensing of proteases in 3D engineered tissues

Dissertação para obtenção do Grau de Doutor em Bioengenharia (MIT)

Non-cell autonomous neuroprotective effect of carbon monoxide : microglia-to-neuron communication

RESUMO: A isquémia cerebral é uma das doenças mais predominantes a nivel mundial, sendo uma das principais causas de mortalidade e invalidez. Parte da propagação de dano no cérebro é causado por inflamação descontrolada, causada principalmente por disfunção da microglia. Desta forma, existe a necessidade de tentar desenvolver estratégias para melhor compreender e modular as acções destas células. O monóxido de carbono (CO), é uma molécula endógena com provas dadas como anti-neuroinflamatório em vários modelos. Assim, o principal objectivo do trabalho foi o estudo do CO como um modulador da acção da microglia, com principal foco dado à comunicação entre estas células e neurónios, tentando entender se existe um efeito neuroprotector por inibição da inflamação. Um protocolo de meio condicionado foi estabelecido usando as linhas celulares BV2 e SH-SY5Y, de microglia e neurónio. A molécula CORM-A1, que liberta expontaniamente CO, foi usada como método de entrega da molécula às celulas. Demonstrámos que o pre-tratamento de células BV2 com CORM-A1 gera neuroprotecção já que reduz a morte celular de neurónios SH-SY5Y quando são incubados com meio condicionado de microglia activada em conjunto com o pró-oxidante t-BHP (tert-butil hidroperóxido). Assim, considerámos que o CO promove neuroprotecção ao inibir as acções inflamatórias da microglia. O papel anti-inflamatório da molécula CORM-A1 foi confirmado quando se verificou que pré-tratamento desta molécula em microglia BV2 limita a secreção de TNF-α mas estimula a secreção de IL-10. Por último, a CORM-A1 induziu a expressão do receptor da microglia CD200R1, molécula que participa na comunicação neurónio-microglia e fundamental para a modulação das acções inflamatórias destas últimas. Em suma, o nosso trabalho reforçou as propriedades anti-neuroinflamatórias do CO e uma capacidade de modular viabilidade neuronal através do seu efeito a nível de comunicação célula-célula. ---------------------------- ABSTRACT: Brain ischemia is a widespread disease worldwide, being one of the main causes of mortality and permanent disability. A portion of the damage that ensues following the ischemic event is caused by unrestrained inflammation, which is mainly orchestrated by exacerbated microglial activity. Hence, developing strategies for modulating microglial inflammation is a major concern nowadays. The endogenous molecule carbon monoxide (CO) has been shown to possess anti-neuroinflammatory properties using in vitro and in vivo approaches. Thus, our objective was to study CO as modulator of microglial activity, in particular in what concerns their communication with neurons, by promoting neuronal viability and limiting inflammatory output of activated microglia. A conditioned media strategy was established with BV2 microglia and SH-SY5Y neurons as cell models. CO-releasing molecule A1 (CORM-A1), a compound that releases CO spontaneously, was used as method of CO delivery to cells. We found that CORM-A1 pre-treatment in BV2 cells yields neuroprotective results, as it limits cell death when SH-SY5Y neurons are challenged with conditioned media from LPS-activated microglia and the pro-oxidant t-BHP (tert-butyl-hydroperoxide). Thus, we assumed carbon monoxide promotes neuroprotection via inhibition of microglial inflammation, displaying a non-cell autonomous role. CORM-A1 pre-treatment limited inflammation by inhibiting BV2 secretion of TNF-α and stimulating IL-10 production. These results reinforce that CO’s anti-inflammatory role confers neuroprotection, as the alterations in these cytokines occur concurrently with the increase in SH-SY5Y viability. Finally, we showed for the first time that carbon monoxide promotes the expression of CD200R1, a microglial receptor involved in neuron-glia communication and modulation of microglia inflammation. Further studies are necessary to clarify this role. Altogether, other than just highlighting CO as an anti-inflammatory and neuroprotective molecule, this work set the foundation for disclosing its involvement in cell-to-cell communication.

Fluid flow regulates stromal cell organization and CCL21 expression in a tissue-engineered lymph node microenvironment.

In the paracortex of the lymph node (LN), T zone fibroblastic reticular cells (TRCs) orchestrate an immune response by guiding lymphocyte migration both physically, by creating three-dimensional (3D) cell networks, and chemically, by secreting the chemokines CCL19 and CCL21 that direct interactions between CCR7-expressing cells, including mature dendritic cells and naive T cells. TRCs also enwrap matrix-based conduits that transport fluid from the subcapsular sinus to high endothelial venules, and fluid flow through the draining LN rapidly increases upon tissue injury or inflammation. To determine whether fluid flow affects TRC organization or function within a 3D network, we regenerated the 3D LN T zone stromal network by culturing murine TRC clones within a macroporous polyurethane scaffold containing type I collagen and Matrigel and applying slow interstitial flow (1-23 microm/min). We show that the 3D environment and slow interstitial flow are important regulators of TRC morphology, organization, and CCL21 secretion. Without flow, CCL21 expression could not be detected. Furthermore, when flow through the LN was blocked in mice in vivo, CCL21 gene expression was down-regulated within 2 h. These results highlight the importance of lymph flow as a homeostatic regulator of constitutive TRC activity and introduce the concept that increased lymph flow may act as an early inflammatory cue to enhance CCL21 expression by TRCs, thereby ensuring efficient immune cell trafficking, lymph sampling, and immune response induction.

Recovering Euclidean deformable models from stereo-motion

In this paper we present a novel structure from motion (SfM) approach able to infer 3D deformable models from uncalibrated stereo images. Using a stereo setup dramatically improves the 3D model estimation when the observed 3D shape is mostly deforming without undergoing strong rigid motion. Our approach first calibrates the stereo system automatically and then computes a single metric rigid structure for each frame. Afterwards, these 3D shapes are aligned to a reference view using a RANSAC method in order to compute the mean shape of the object and to select the subset of points on the object which have remained rigid throughout the sequence without deforming. The selected rigid points are then used to compute frame-wise shape registration and to extract the motion parameters robustly from frame to frame. Finally, all this information is used in a global optimization stage with bundle adjustment which allows to refine the frame-wise initial solution and also to recover the non-rigid 3D model. We show results on synthetic and real data that prove the performance of the proposed method even when there is no rigid motion in the original sequence

Online Robust 3D Mapping Using Structure from Motion Cues

This paper presents a complete solution for creating accurate 3D textured models from monocular video sequences. The methods are developed within the framework of sequential structure from motion, where a 3D model of the environment is maintained and updated as new visual information becomes available. The camera position is recovered by directly associating the 3D scene model with local image observations. Compared to standard structure from motion techniques, this approach decreases the error accumulation while increasing the robustness to scene occlusions and feature association failures. The obtained 3D information is used to generate high quality, composite visual maps of the scene (mosaics). The visual maps are used to create texture-mapped, realistic views of the scene

Restitución 3D de la topografía de la antigua ciudad de Tarraco en un entorno SIG: propuestas metodológicas y primeros resultados

Durante los últimos años el Institut Català d’Arquelogia Clàssica, el Museu d’Història de Tarragona, contando con la colaboración de la Generalitat de Catalunya, han desarrallado el proyecto Planimetría Arqueológica de Tárraco, destinado a la elaboración de una planta arqueológica global en la cual se recogieran intervenciones y noticias referentes a los hallazgos arqueológicos existentes. Este trabajo fue publicado utilizando como plataforma de trabajo un SIG construido para tal fin (Macias et al. 2007). Sin embargo, un problema de difícil solución arqueológica venía dado por las transformaciones urbanísticas de la ciudad, sufridas en su mayor parte a lo largo de los siglos XIX y XX. Éstas habían provocado la pérdida irremediable de gran parte de la elevación que acogiera la ciudad romana, cambiando substancialmente su aspecto original. Ante esta situación y como proyecto paralelo a la realización de la Planimetría Arqueológica de Tarragona se plantearon formas de cubrir este vacío. Se presenta en esta comunicación una propuesta metodológica para la reconstrucción de los grandes «vacíos topográficos » originados por la evolución urbanística de Tarragona mediante la obtención e integración en un SIG de diversos tipos de información documental. En estas zonas rebajadas no resulta posible la obtención de información estratigráfica y arqueológica, por lo que es imprescindible la definición de vías metodológicas alternativas basadas en la extrapolación de datos extraídos de la cartografía histórica, panorámicas del XVI o fotografías tomadas en los siglos XIX y XX. Esta técnica permite aplicar los resultados obtenidos en los nuevos análisis interpretativos, complementando así la interpretación arqueológica de la topografía urbana de la ciudad romana. A partir de esta información, y aplicando funciones y técnicas de interpolación propias de un GIS, se propone aquí un modelo de relieve de la ciudad de Tarraco.

The Application of 3D Reconstruction Techniques in the Analysis of Ancient Tarraco’s Urban Topography

The widespread implementation of GIS-based 3D topographical models has been a great aid in the development and testing of archaeological hypotheses. In this paper, a topographical reconstruction of the ancient city of Tarraco, the Roman capital of the Tarraconensis province, is presented. This model is based on topographical data obtained through archaeological excavations, old photographic documentation, georeferenced archive maps depicting the pre-modern city topography, modern detailed topographical maps and differential GPS measurements. The addition of the Roman urban architectural features to the model offers the possibility to test hypotheses concerning the ideological background manifested in the city shape. This is accomplished mainly through the use of 3D views from the main city accesses. These techniques ultimately demonstrate the ‘theatre-shaped’ layout of the city (to quote Vitrubius) as well as its southwest oriented architecture, whose monumental character was conceived to present a striking aspect to visitors, particularly those arriving from the sea.

Establishment and characterization of models of chemotherapy resistance in colorectal cancer: Towards a predictive signature of chemoresistance.

Current standard treatments for metastatic colorectal cancer (CRC) are based on combination regimens with one of the two chemotherapeutic drugs, irinotecan or oxaliplatin. However, drug resistance frequently limits the clinical efficacy of these therapies. In order to gain new insights into mechanisms associated with chemoresistance, and departing from three distinct CRC cell models, we generated a panel of human colorectal cancer cell lines with acquired resistance to either oxaliplatin or irinotecan. We characterized the resistant cell line variants with regards to their drug resistance profile and transcriptome, and matched our results with datasets generated from relevant clinical material to derive putative resistance biomarkers. We found that the chemoresistant cell line variants had distinctive irinotecan- or oxaliplatin-specific resistance profiles, with non-reciprocal cross-resistance. Furthermore, we could identify several new, as well as some previously described, drug resistance-associated genes for each resistant cell line variant. Each chemoresistant cell line variant acquired a unique set of changes that may represent distinct functional subtypes of chemotherapy resistance. In addition, and given the potential implications for selection of subsequent treatment, we also performed an exploratory analysis, in relevant patient cohorts, of the predictive value of each of the specific genes identified in our cellular models.

3D-tuoteyhdistelmämallien ja mittakuvien tuottaminen PDM-järjestelmästä

Tämä diplomityö on selvitystyö mittakuvien ja kolmiulotteisten CAD-mallien tuottamisesta. Mittakuvat ja 3D-CAD-mallit halutaan Neles-tuotemerkin omaaville tuotteille. Olennaisena osana työssä on tuotetiedonhallintajärjestelmä AtonPDM, koska mittakuvia ja malleja toivottaisiin hallittavan AtonPDM-järjestelmällä. Työ tehdään Metso Automationin (MA) Flow Control (FC) –liiketoimintalinjalle. Nykyiset mittakuvat aiheuttavat ongelmia sekä MA:ssa että asiakkaille. MA:ssa mittakuvien tekeminen kestää kauemmin kuin asiakas toivoisi. Nykyisen mittakuvaohjelmiston riittämättömät ominaisuudet aiheuttavat lisätyötä mittakuvien valmistuksessa. Asiakkaille mittakuvien viivästyminen on suunnittelua hidastava tekijä. Mittakuvissa olevat virheet ja puutteet vaikeuttavat asiakkaan suunnittelutyötä ja saattavat päästä läpi tarkatuksien myös loppusuunnitelmiin, jolloin seurauksena voi olla rahallisia menetyksiä. Tämän päivän putkistosuunnittelu hoidetaan suurimmaksi osaksi 3D-CADohjelmistoilla. Suunnittelua helpotetaan ja nopeutetaan valmiilla komponenttien 3Dmalleilla, joihin on liitetty tuotetietoa. Työssä on haastateltu MA:n henkilökuntaa, CAD-järjestelmien toimittajia ja asiakkaita. Haastattelut ovat työn tärkein tiedonlähde. Teoriatietoa on selvitetty kirjoista, lehdistä ja internetistä. Teoriaosiossa käydään läpi tuotetiedonhallinta (Product Data Management, PDM), tietokanta ja parametrinen mallintaminen. Työn lopputuloksena on pyritty saamaan kuvaus siitä miten tuotetaan 3D-CAD-mallit ja mittakuvat Neles-tuotteista käyttäen tiedonlähteenä AtonPDM:ää ja millaisia 3Dmallien ja mittakuvien tulee olla. Malleja ja mittakuvia tuottavaan järjestelmään on etsitty ratkaisuja CAD-ohjelmistoista. Ratkaisuja on verrattu keskenään ominaisuuksien, joustavuuden ja AtonPDM yhteensopivuuden perusteella.

Visualització de models fusionats

L’objectiu principal d’aquest projecte era implementar la visualització 3D de models fusionats i aplicar totes les tècniques possibles per realitzar aquesta fusió. Aquestes tècniques s’integraran en la plataforma de visualització i processament de dades mèdiques STARVIEWER. Per assolir l’ objectiu principal s’ han definit els següents objectius específics:1- estudiar els algoritmes de visualització de models simples i analitzar els diferents paràmetres a tenir en compte. 2- ampliació de la tècnica de visualització bàsica seleccionada per tal de suportar els models fusionats. 3- avaluar i compar tots els mètodes implementats per poder determinar quin ofereix les millors visualitzacions

Recovering Euclidean deformable models from stereo-motion

In this paper we present a novel structure from motion (SfM) approach able to infer 3D deformable models from uncalibrated stereo images. Using a stereo setup dramatically improves the 3D model estimation when the observed 3D shape is mostly deforming without undergoing strong rigid motion. Our approach first calibrates the stereo system automatically and then computes a single metric rigid structure for each frame. Afterwards, these 3D shapes are aligned to a reference view using a RANSAC method in order to compute the mean shape of the object and to select the subset of points on the object which have remained rigid throughout the sequence without deforming. The selected rigid points are then used to compute frame-wise shape registration and to extract the motion parameters robustly from frame to frame. Finally, all this information is used in a global optimization stage with bundle adjustment which allows to refine the frame-wise initial solution and also to recover the non-rigid 3D model. We show results on synthetic and real data that prove the performance of the proposed method even when there is no rigid motion in the original sequence

Online Robust 3D Mapping Using Structure from Motion Cues

This paper presents a complete solution for creating accurate 3D textured models from monocular video sequences. The methods are developed within the framework of sequential structure from motion, where a 3D model of the environment is maintained and updated as new visual information becomes available. The camera position is recovered by directly associating the 3D scene model with local image observations. Compared to standard structure from motion techniques, this approach decreases the error accumulation while increasing the robustness to scene occlusions and feature association failures. The obtained 3D information is used to generate high quality, composite visual maps of the scene (mosaics). The visual maps are used to create texture-mapped, realistic views of the scene

Efficient 3D scene modeling and mosaicing

El modelat d'escenes és clau en un gran ventall d'aplicacions que van des de la generació mapes fins a la realitat augmentada. Aquesta tesis presenta una solució completa per a la creació de models 3D amb textura. En primer lloc es presenta un mètode de Structure from Motion seqüencial, a on el model 3D de l'entorn s'actualitza a mesura que s'adquireix nova informació visual. La proposta és més precisa i robusta que l'estat de l'art. També s'ha desenvolupat un mètode online, basat en visual bag-of-words, per a la detecció eficient de llaços. Essent una tècnica completament seqüencial i automàtica, permet la reducció de deriva, millorant la navegació i construcció de mapes. Per tal de construir mapes en àrees extenses, es proposa un algorisme de simplificació de models 3D, orientat a aplicacions online. L'eficiència de les propostes s'ha comparat amb altres mètodes utilitzant diversos conjunts de dades submarines i terrestres.

Synthesis and structure-activity relationship studies of CD4 down-modulating cyclotriazadisulfonamide (CADA) analogues

HIV attachment via the CD4 receptor is an important target for developing novel approaches to HIV chemotherapy. Cyclotriazadisulfonamide (CADA) inhibits HIV at submicromolar levels by specifically down-modulating cell-surface and intracellular CD4. An effective five-step synthesis of CADA in 30% overall yield is reported. This synthesis has also been modified to produce more than 50 analogues. Many tail-group analogues have been made by removing the benzyl tail of CADA and replacing it with various alkyl, acyl, alkoxycarbonyl and aminocarbonyl substituents. A series of sidearm analogues, including two unsymmetrical compounds, have also been prepared by modifying the CADA synthesis, replacing the toluenesulfonyl sidearms with other sulfonyl groups. Testing 30 of these compounds in MT-4 cells shows a wide range of CD4 down-modulation potency, which correlates with ability to inhibit HIV-1. Three-dimensional quantitative structure-activity relationship (3D-QSAR) models were constructed using comparative molecular field analysis (CoMFA) and comparative molecular similarity indices analysis (CoMSIA) approaches. The X-ray crystal structures of four compounds, including CADA, show the same major conformation of the central 12-membered ring. The solid-state structure of CADA was energy minimized and used to generate the remaining 29 structures, which were similarly minimized and aligned to produce the 3D-QSAR models. Both models indicate that steric bulk of the tail group, and, to a lesser extent, the sidearms mainly determine CD4 down-modulation potency in this series of compounds.