Cancer is the second largest mortal disease in Sweden, and high efforts are made to develop the treatment of cancer. One of the main treatment methods is radiotherapy, which uses ionizing radiation to damage the cancerous cells. This has the chance of stopping the cell reproduction, and the goal is to reduce the tumor and stop the tumor growth. The most common forms of radiotherapy uses external beams to irradiate the tumor. In intensity modulated radiotherapy, IMRT, the beam fluences are optimized to give a highly conformal dose, i.e. a dose distribution which is restricted to the tumor and has low dose values outside of the tumor. A conformal dose is necessary to spare healthy tissue and sensitive organs, and thus keep the side-effects of the treatment at an acceptable level. The optimized beam shapes are created using a multileaf collimator, MLC. Finding the leaf positions and dose levels is formulated as a problem in the framework of mathematical optimization. Currently, one of the limitations in delivering conformal dose is due to patient movement during the treatment. In IMRT, the beams are delivered by consecutive segments, and the exact pairing of the segments with the patient position will have an impact on the delivered dose. This is called the interplay effect, and can cause both underdosage of the tumor and overdosage of the surrounding tissue. There are methods of mitigating the interplay effect. For example, the beam could be restricted to a single phase of the motion by repeatedly turning it on and off. This is known as gating. However, gating and many other interplay mitigation techniques lead to prolonged treatment times, which decreases the clinical throughput, causes higher patient discomfort and gives higher uncertainties in the delivered dose. This makes it desirable to find methods which avoid prolonged treatment times, while still giving highly conformal doses. Ideally, the best method would be to have a beam which follows any target movement. This idea is known as target tracking. In this thesis, an optimization method is suggested which includes the interplay effect in the treatment optimization. Two main treatment strategies are proposed. The method which is simplest to implement clinically is to create plans which are robust against uncertainties in the times for the patient motion. The resulting doses are found to give acceptable target covering where similar, conventional plans give a significant target underdose. To further increase the conformality of the doses, a non-robust method paired with gating technology is suggested. This method can effectively be seen as a target tracking method, and has the possibility to give highly conformal doses under acceptable treatment times.

Base teórica: O câncer de pulmão é uma doença com alta incidência e mortalidade, cujo prognóstico permanece discreto apesar do melhor entendimento da doença nas últimas décadas. A radioterapia tem papel terapêutico em todos os estágios da doença. A expressão da cofilina-1, uma proteína relacionada à mobilidade celular, determinou maior radiossensibilidade a células de adenocarcinoma brônquico em estudos in vitro, porém pior sobrevida em estádios iniciais Objetivo: Avaliar se a expressão da cofilina-1 interfere na sobrevida e no controle local em pacientes com câncer de pulmão submetidos a tratamento com radioterapia definitiva Métodos: Foram avaliados pacientes com câncer de pulmão não pequenas células, com estádios I–IV, que receberam radioterapia exclusiva ou combinada com quimioterapia, dirigida à neoplasia brônquica, na unidade de radioterapia do HCPA, nos anos de 2009 a 2015. Todos os pacientes tiveram a expressão de cofilina-1 aferida e foram distribuídos conforme os níveis de expressão de cofilina-1 utilizando-se de protocolo específico. Os prontuários foram avaliados retrospectivamente para calcular a sobrevida mediana. A progressão foi verificada através de avaliação de tomografias de tórax de controle. Resultados: Foram avaliados 45 pacientes neste estudo. A sobrevida mediana de todos os pacientes foi de 11,3 meses e a sobrevida global em 5 anos de 17,3%. Pacientes com expressão média ou alta de cofilina-1 tiveram maior mortalidade quando comparados com pacientes com baixa expressão (respectivamente, HR 1,628, IC95 1,137-8,287 e HR 1,59 IC95 1,105-7,342). Não houve diferença significantemente estatística entre controle local e expressão de cofilina-1. Conclusão: A expressão de cofilina-1 está associada à sobrevida em pacientes com carcinoma brônquico tratados com radioterapia e existe uma tendência a melhor controle local com baixa expressão. Nossos resultados sugerem um novo campo a ser explorado no manejo do carcinoma de pulmão localmente avançado, utilizando-se dos níveis de cofilina-1. / Background: Lung cancer is a disease with high incidence and mortality, whose prognosis remains poor despite a better understanding of the disease in the last decades. Radiotherapy plays a therapeutic role in all stages of disease. The expression of cofilin-1, a protein related to cellular mobility, determined greater radiosensitivity to lung adenocarcinoma cells in in vitro studies, but worse survival at initial stages. Objective: To evaluate if the expression of cofilin-1 modified survival and local control in lung cancer patients submitted to definitive treatment with radiotherapy. Methods: Patients with non-small cell lung cancer with stage IIV who received radiotherapy alone or combined with chemotherapy for lung cancer at the HCPA radiotherapy unit from 2009 to 2015 were evaluated. All patients had the expression of measured cofilin-1 evaluated and were distributed by cofilin-1 expression according to specific protocol. The medical records were retrospectively evaluated to estimate median survival. The progression was verified through evaluation of control chest tomography. Results: 45 patients were assessed in this study. The median survival of all patients was 11.3 months and the 5-year overall survival was 17.3%. Patients with medium or high expression of cofilin-1 had higher mortality rates when compared to patients with low expression (HR, 1,628, CI95, 1,137-8,287 and HR, 1.59, CI95, 1,105-7,342). There was no statistically significant difference between local control and cofilin-1 expression. Conclusion: cofilin-1 expression is associated with survival in patients with lung cancer treated with radiotherapy and there is a tendency for better local control with low CFL1 expression. Our results suggest a new field to be explored in the management of locally advanced lung carcinoma, using cofilin-1 expression levels.

Base teórica: O câncer de pulmão é uma doença com alta incidência e mortalidade, cujo prognóstico permanece discreto apesar do melhor entendimento da doença nas últimas décadas. A radioterapia tem papel terapêutico em todos os estágios da doença. A expressão da cofilina-1, uma proteína relacionada à mobilidade celular, determinou maior radiossensibilidade a células de adenocarcinoma brônquico em estudos in vitro, porém pior sobrevida em estádios iniciais Objetivo: Avaliar se a expressão da cofilina-1 interfere na sobrevida e no controle local em pacientes com câncer de pulmão submetidos a tratamento com radioterapia definitiva Métodos: Foram avaliados pacientes com câncer de pulmão não pequenas células, com estádios I–IV, que receberam radioterapia exclusiva ou combinada com quimioterapia, dirigida à neoplasia brônquica, na unidade de radioterapia do HCPA, nos anos de 2009 a 2015. Todos os pacientes tiveram a expressão de cofilina-1 aferida e foram distribuídos conforme os níveis de expressão de cofilina-1 utilizando-se de protocolo específico. Os prontuários foram avaliados retrospectivamente para calcular a sobrevida mediana. A progressão foi verificada através de avaliação de tomografias de tórax de controle. Resultados: Foram avaliados 45 pacientes neste estudo. A sobrevida mediana de todos os pacientes foi de 11,3 meses e a sobrevida global em 5 anos de 17,3%. Pacientes com expressão média ou alta de cofilina-1 tiveram maior mortalidade quando comparados com pacientes com baixa expressão (respectivamente, HR 1,628, IC95 1,137-8,287 e HR 1,59 IC95 1,105-7,342). Não houve diferença significantemente estatística entre controle local e expressão de cofilina-1. Conclusão: A expressão de cofilina-1 está associada à sobrevida em pacientes com carcinoma brônquico tratados com radioterapia e existe uma tendência a melhor controle local com baixa expressão. Nossos resultados sugerem um novo campo a ser explorado no manejo do carcinoma de pulmão localmente avançado, utilizando-se dos níveis de cofilina-1. / Background: Lung cancer is a disease with high incidence and mortality, whose prognosis remains poor despite a better understanding of the disease in the last decades. Radiotherapy plays a therapeutic role in all stages of disease. The expression of cofilin-1, a protein related to cellular mobility, determined greater radiosensitivity to lung adenocarcinoma cells in in vitro studies, but worse survival at initial stages. Objective: To evaluate if the expression of cofilin-1 modified survival and local control in lung cancer patients submitted to definitive treatment with radiotherapy. Methods: Patients with non-small cell lung cancer with stage IIV who received radiotherapy alone or combined with chemotherapy for lung cancer at the HCPA radiotherapy unit from 2009 to 2015 were evaluated. All patients had the expression of measured cofilin-1 evaluated and were distributed by cofilin-1 expression according to specific protocol. The medical records were retrospectively evaluated to estimate median survival. The progression was verified through evaluation of control chest tomography. Results: 45 patients were assessed in this study. The median survival of all patients was 11.3 months and the 5-year overall survival was 17.3%. Patients with medium or high expression of cofilin-1 had higher mortality rates when compared to patients with low expression (HR, 1,628, CI95, 1,137-8,287 and HR, 1.59, CI95, 1,105-7,342). There was no statistically significant difference between local control and cofilin-1 expression. Conclusion: cofilin-1 expression is associated with survival in patients with lung cancer treated with radiotherapy and there is a tendency for better local control with low CFL1 expression. Our results suggest a new field to be explored in the management of locally advanced lung carcinoma, using cofilin-1 expression levels.

Este estudo avaliou, in vitro, a influência da radioterapia na resistência de união (RU) e na interface adesiva entre diferentes cimentos resinosos e dentina radicular. Sessenta caninos superiores foram distribuídos em 2 grupos (n=30) de acordo com a irradiação: não irradiados e irradiados. Os dentes do grupo irradiado foram submetidos à radioterapia com raios-X de 6 MV em frações de 2 Gy, com 30 ciclos, perfazendo 60 Gy. Os dentes foram seccionados para obtenção de raízes com 16 mm de comprimento, sendo realizado em seguida o preparo biomecânico com instrumento Reciproc R50 e obturação pela técnica de condensação lateral com cimento a base resina epóxica. Cada grupo foi subdividido de acordo com o cimento resinoso utilizado para a cimentação dos pinos de fibra de vidro (n=10): RelyX U200; Panavia F 2.0; RelyX ARC. Os pinos foram submetidos a tratamento de superfície com ácido fluorídrico 10% e cimentados de acordo com as recomendações de cada fabricante. Após a cimentação dos pinos, os dentes foram seccionados transversalmente em slices de 1 mm de espessura, obtendo-se 3 slices de cada terço da raiz. Os slices mais cervicais de cada terço foram utilizados para avaliar a RU, por meio do teste de push-out com velocidade de 0,5 mm/min, e posterior análise do padrão de falha em estereomicroscópio. Os slices mais apicais de cada terço foram selecionados para análise da interface cimento/dentina em MEV com aumentos de 100, 1000, 2000 e 4000X. Os dados de RU e adaptação da interface cimento/dentina foram submetidos à análise estatística pelos testes de ANOVA e Tukey, e Kruskal-Wallis e Duns, respectivamente. Os espécimes irradiados apresentaram menores valores de RU (8,23&plusmn;4,26) comparados aos dentes não irradiados (11,88&plusmn;6,42) (p<0,00001). Quanto aos cimentos resinosos, o RelyX U200 apresentou maiores valores na RU (15,17&plusmn;5,89) comparado aos cimentos RelyX ARC (7,68&plusmn;4,22) (p<0,0001) e Panavia F 2.0 (7,32&plusmn;2,71) (p<0,0001). O terço cervical apresentou maiores valores de RU (13,08&plusmn;6,10) comparado aos terços médio (9,72&plusmn;5,03) e apical (7,38&plusmn;4,53) (p<0,0001). O padrão de falhas mostrou ocorrência de falhas coesivas na dentina para os espécimes irradiados. Na análise da interface cimendo/dentina por MEV, observou-se maior desadaptação nos dentes submetidos à radioterapia. Em relação aos cimentos resinosos, foi observada maior adaptação com RelyX U200 e RelyX ARC comparados à Panavia F 2.0. Na análise qualitativa em MEV observou-se presença de fraturas e microfraturas na dentina radicular e menor presença de fibras colágenas em dentes irradiados. Para o cimento resinoso RelyX U200 e Panavia F 2.0 observou-se interface justaposta do cimento com a dentina radicular em dentes irradiados e não irradiados, sendo que para o cimento resinoso RelyX ARC foi observada formação de camada híbrida e tags de maneira similar para dentes irradiados e não irradiados. Concluiu-se que a radioterapia resultou na redução da resistência de união e na maior desadaptação da interface cimento resinoso/dentina radicular e que o cimento resinoso autoadesivo se apresentou como melhor alternativa para cimentação de pinos de fibra de vidro em dentes irradiados. / This in vitro study, evaluated the influency of radiotherapy on bond strength (BS) and adhesive interface between different resin cements and root dentin. Sixty maxillary canines were selected and distributed into two groups (n = 30) according to the irradiation protocol: non-irradiated and irradiated. The irradiated group were submitted to X-ray radiotherapy of 6 MV in fractions of 2 Gy, with 30 cycles, until complete 60 Gy. The teeth were sectioned to obtain 16 mm of root length, followed by biomechanical preparation with R50 reciproc instrument and obturation using the lateral condensation technique with epoxy resin-based sealer. Then, each group was subdivided according to the resin cement used for the glass fiber post cementation (n = 10): RelyX U200, Panavia F 2.0 and RelyX ARC. The posts were submitted to surface treatment with 10% hydrofluoric acid, and then cemented according to the manufacturer instructions. After the posts cementation, the teeth were sectioned transversely into 1 mm thick slices, and 3 slices were obtained from each root third. The most cervical slice of each third was used to evaluate the BS by the push-out test at 0.5 mm/min of velocity and the failure pattern was analyzed using stereomicroscopy. The most apical slice of each third was selected for SEM analysis, which were prepared and metalized. The dentin/cement interface analysis was performed at the following magnification: 100, 1000, 2000 and 4000X. The BS data and dentin/cement interface adaptation were submitted to statistical analysis by ANOVA, Tukey and Kruskal-Wallis, and Duns tests respectively. The irradiated specimens had lower BS values (8.23 &plusmn; 4.26) compared to non-irradiated group (11.88 &plusmn; 6.42) (p <0.00001). Regarding the resin cements, the RelyX U200 showed the higher values in BS (15.17 &plusmn; 5.89) compared to RelyX ARC (7.68 &plusmn; 4.22) and Panavia F 2.0 (7 , 32 &plusmn; 2.71) (p <0.0001). In addition, the cervical third presented higher BS values (13.08 &plusmn; 6.10) when compared to the middle (9.72 &plusmn; 5.03) and apical (7.38 &plusmn; 4.53) thirds (p <0.0001). The failure pattern showed cohesive failures in root dentin of the irradiated specimens. In the SEM dentin/cement interface analysis, a greater misadaptation was observed for the teeth submitted to radiotherapy. Regarding the resin cements, a better adaptation was observed for the RelyX U200 and RelyX ARC compared to the Panavia F 2.0. In the SEM qualitative analysis of the irradiated teeth, was possible to observe the presence of fractures and microfractures in the root dentin and a small number of collagen fibers. The resin cements RelyX U200 and Panavia F 2.0 showed a juxtaposed interface of the cement with the root dentin in irradiated and non-irradiated teeth, being that for the RelyX ARC cement the presence of hybrid layer and tags was similar for irradiated and non-irradiated groups. It was concluded that the radiotherapy promoted reduction of the bond strength and increase the gaps in the dentin/resin cement interface and the self-adhesive resin cement is the best alternative for glass fiber posts cementation in irradiated teeth.

La radiothérapie est souvent utilisée pour contrôler la progression d'un cancer. Cependant, la mauvaise spécificité de ciblage de la plupart des techniques de radiothérapie peut entraîner une réponse clinique ambiguë. Une stratégie alternative - et complémentaire - est d'utiliser des matériaux possédant un numéro atomique élevé et qui peuvent ainsi agir en synergie avec les rayonnements ionisants pour améliorer le ratio thérapeutique de la radiothérapie. Dans ce contexte, une nanoparticule (NP) théranostique à base de gadolinium (Gd) est particulièrement adaptée pour fournir simultanément une plus grande précision en Imagerie par Résonance Magnétique (IRM) et une meilleure efficacité en radiothérapie clinique. Au cours de cette thèse, nous avons étudié d'un point de vue préclinique la pharmacocinétique et le métabolisme de ces NP chez des rongeurs et des primates non humains afin d'élucider leurs voies d'élimination et de calculer la dose sans effet nocif observé (NOAEL). De plus, nous avons démontré la capacité d'imagerie et de thérapie de ces particules sur un modèle de souris porteuses de mélanome cérébral, ceci afin d'appuyer le potentiel des NP pour la radiothérapie guidée par IRM en clinique. Ces travaux de thèse - ainsi que des résultats précédents - ont contribué au début d'un essai clinique actuellement en cours / Combinations of chemotherapy and radiotherapy are often used to control cancer progression. However, the poor targeting specificity of most chemotherapies and radiotherapies can cause toxicity and ambiguous clinical response. In particular, dose escalation in radiotherapy inevitably increases radiation exposure for some surroundings normal tissues and organs, putting them at risk for debilitating damage. An alternative – and complementary – strategy is the use of materials with high atomic numbers (Z) that strongly interact with low energy photons to produce photoelectrons and Auger electrons In this context, a new efficient type of gadolinium (Gd)-based theranostic agent (AGuIX®) has recently been developed by the team of Prof. Tillement for MRI-guided radiotherapy. AGuIX® is a 3-nm size nanoparticles of 9 kDa, consist of a polysiloxane network surrounded by Gd chelates. In this thesis, we investigated the elimination kinetics of AGuIX nanoparticle’s (NPs) from sub-cellular to whole organ scale using original and complementary techniques. This combination of techniques allows the exact mechanism of AGuIX NPs elimination to be elucidated. We reported the preclinical pharmacokinetics and toxicology studies of intravenous AGuIX NPs administration in healthy and atherosclerosis non-human primates (NHP), the goal of which is to demonstrate the safety of AGuIX NPs, in particular, for pre-clinical evaluation. Subsequently, we performed experimental and theoretical studies to investigate the radiosensitization of AGuIX NPs, in particular with B16F10 mouse melanoma as a model for brain metastases. After, we implemented experimental and theoretical studies to precisely understand the mechanism of this radiosensitization, we suggest additional mechanism, potentially caused by chemical and biological effects induced by the combination of Gd and radiation (i.e. high yield of radicals formation and combination, and bystander effect)

La Thérapie Photodynamique ou PDT est une méthode de traitement principalement anti-cancéreux. Elle est basée sur l’activation par la lumière de molécules photosensibles, appelés photosensibilisateurs, non toxiques à l’obscurité. Après excitation lumineuse et en présence d’oxygène, elles génèrent des espèces réactives de l’oxygène dont l’oxygène singulet, qui engendrent des réactions de photo-oxydation entraînant la mort cellulaire. Cette méthode est cliniquement appliquée en France et dans plusieurs pays du monde mais présente toutefois certaines limites comme la faible profondeur de pénétration de la lumière dans les tissus. Elle ne peut donc s’appliquer qu’aux tumeurs de surface telles que les kératoses actiniques et aux tumeurs de petite taille accessibles à la lumière. Pour traiter les tumeurs profondes par PDT, nous proposons une stratégie innovante basée sur l’utilisation de rayons X, très connus pour leur pouvoir pénétrant. Comme la plupart des photosensibilisateurs ne sont pas radiosensibles, ils ne peuvent être excités directement par les rayons X. Les récentes avancées en nanotechnologie nous ont amené à envisager une stratégie thérapeutique novatrice pouvant combiner les principes de la PDT et la radiothérapie via l’élaboration de scintillateurs nanoparticulaires excitables par rayons X. Le concept consiste à utiliser des nanoparticules contenant dans leur cœur un scintillateur qui, excité par des rayons X, peut émettre des photons, qui, à leur tour, sont réabsorbés par le photosensibilisateur lui-même conjugué à la nanoparticule. C’est dans ce contexte que nous avons développé des nanoparticules Gd2O2S : Eu3+, possédant dans leur cœur du gadolinium (agent permettant le rehaussement positif du signal IRM) dopé avec de l’europium ou du terbium (pour une l’absorption des RX) et conjugués à des photosensibilisateurs (porphyrine, chlorine zinguée et phtalocyanine zinguée). D’autres nanoparticules TbO3, de plus petite taille, possédant dans leur cœur de l’oxyde de terbium, enrobé d’une couche de polysiloxane dans laquelle est incorporée de la porphyrine ont aussi été synthétisées et étudiées. Ces nanoparticules offrent à la fois une possibilité d’effet radiothérapie, d’effet PDT, tout en offrant la possibilité de visualiser les tumeurs par IRM. Nous avons mis au point un protocole de synthèse permettant d’obtenir la porphyrine avec de bons rendements tout en limitant le temps de synthèse et les étapes de purification. Les synthèses et purifications des nanoparticules ont été optimisées. Les caractérisations physico-chimiques et photophysiques des nanoparticules ont été réalisées, en particulier leur capacité à produire de l’oxygène singulet. L’efficacité du transfert d’énergie entre les nanoparticules et les photosensibilisateurs dans le but d’obtenir un effet PDT a été évaluée et des essais préliminaires sous excitation par rayons X ont été entrepris / PDT or photodynamic therapy is a method mainly used against cancer. PDT is based on the activation by light of photosensitive molecules, called photosensitizers, non-toxic in the dark. After excitation by light in the presence of oxygen, the photosensitizers generate reactive oxygen species including singlet oxygen, which lead to photo-oxidation reactions and cell death. This method is clinically applied in France and in several countries of the world but still has some limitations such as the low depth of light penetration in tissues. It therefore can be applied only to surface tumors such as actinic keratoses and small tumors accessible to laser light. To treat deep tumors by PDT, we propose an innovative strategy based on the use of X-rays known for their penetrating power. Since most photosensitizers are not radiosensitive, they cannot be excited by X-rays. Recent advances in nanotechnology have led us to consider a novel therapeutic strategy that can combine both the principles of PDT and radiotherapy through the development of scintillator containing nanoparticles excitable by X-rays. The concept is to use the scintillator included in the nanoparticle’s core which, excited by X-rays, can emit photons, which, in turn, can be re-absorbed by the photosensitizer itself conjugated to the nanoparticles. It is in this context that we developed nanoparticles of Gd2O2S:Eu3+, with in their core gadolinium (which is a MRI positive enhancement agent) doped with europium or terbium (for RX absorption) and coupled with photosensitizers (porphyrin, zinc chlorin and zinc phthalocyanine). We have developed a synthesis protocol for obtaining the porphyrin in good yields while limiting the time of synthesis and purification steps. The syntheses and purifications of nanoparticles were optimized. The physicochemical and photophysical characterization of the nanoparticles were performed, in particular their ability to produce singlet oxygen. Other Tb2O3 nanoparticles of smaller size, coated with a polysiloxane layer coupled to porphyrin have also been synthesized and studied. These nanoparticles offer both an opportunity for radiotherapy effect of PDT effect, while providing the ability to be detected by MRI. The efficiency of energy transfer between the nanoparticles and the photosensitizers in order to obtain a PDT effects was assessed and preliminary tests under X-ray excitation were undertaken

Le but principal de ce projet est de préparer l’implantation clinique du système Clarity qui utilise une sonde ultrasonore pour visualiser l’anatomie interne du patient. Ce système est utilisé pour les cas de prostate et nécessite d’être adapté pour les cas de poumon. L’utilité de ce système est de suivre un substitut afin d’inférer la position d’une tumeur pulmonaire. L’hypothèse de cette étude est qu’un substitut interne serait mieux corrélé avec une tumeur pulmonaire que le seraient des marqueurs externes. Les sous-objectifs sont : 1) aborder l’adaptation du montage pour faire des acquisitions sur des patients ; 2) explorer la performance des algorithmes de détection de mouvements ainsi que des métriques de qualité d’image sur des images US et ciné IRM; 3) démontrer que la corrélation entre un substitut interne et une structure pulmonaire est plus grande que celle avec un substitut externe. Pour les acquisitions d’images US, la sonde est placée sur les volontaires et fixée à la table de traitement à l’aide d’un bras mécanique. Il a été démontré qu’une pression insuffisante peut causer une perte de signal dû à la forme curviligne de la sonde. La diminution de la moyenne des intensités de l’image et de l’écart-type confirme une perte de signal lors d’amplitudes respiratoires élevées justifiée par une perte de contact entre la sonde et la peau malgré la fixation de la sonde. Entre les algorithmes de corrélation croisée normalisée (NCC), d’erreur moyenne quadratique (RMS) et de flux optique, la méthode NCC semble la plus robuste pour suivre le substitut interne (structure dans le foie) dans les images IRM pour 5/9 volontaires sains ( = 0, 050). Cette méthode est utilisé présentement pour les cas de prostate. Le flux optique s’est montré plus efficace pour des cas spécifiques ce qui démontre l’intérêt d’adapter l’algorithme pour les cas de poumons. Enfin, il a été démontré sur les images IRM qu’un substitut interne au niveau du foie est plus efficace pour la majorité des volontaires (8/9) en comparaison avec un marqueur sur la peau placé dans la région abdominale. Le marqueur abdominal possède une meilleure corrélation qu’un marqueur thoracique (9/9) illustrant l’importance du positionnement d’un marqueur externe pour le suivi d’une tumeur pulmonaire. / The main objective of this thesis is to prepare the clinical implementation of the Clarity ultrasound system for indirect lung tumours tracking using a surrogate. It is currently used for motion management during prostate treatments and requires adaptation. Our hypothesis is that an internal marker would have a better correlation with the tumour’s position than an external surrogate. The sub-objectives are : 1) test different setups for the image acquisition on patients ; 2) explore the algorithms’ performance for motion detection as well as the image quality metrics on US and dynamic MRI images ; 3) evaluate the correlation between surrogates and a lung structure to determine which performs best. The ultrasound probe is fixed on the treatment couch for the acquisition on healthy volunteers using a mechanical arm. Low pressure on the patient’s skin results in a loss of signal due to the curvilinear shape of the probe. We observed a loss of contact between the probe and the volunteers’ skin due to ample movements causing a deterioration of the image quality. We tested three different motion detection algorithms on dynamic MRI images : normalized cross-correlation (NCC), root mean square error (RMS) and optical flow. The NCC algorithm is the most robust out of the three for 5/9 volunteers for the internal surrogate tracking ( < 0.050). In specific cases, the optical flow method performed better indicating an interest in developping a new algorithm for indirect lung tracking. Finally, the correlation between the surrogates and a lung structure were calculated using the MRI images. The internal surrogate inside the liver was proven more efficient for indirect lung tumour tracking for 8/9 volunteers. External markers give a greater prediction error. It has also been shown that the positioning of the external marker on the patient’s skin impacts the correlation. The abdominal marker is better than the thoracic one for all the volunteers.

<p>Vitamin E is a natural lipophilic antioxidant contains eight structurally related forms, i.e., α-, β-, γ-, δ-tocopherols (αT, βT, γT, and δT) and corresponding tocotrienols. Recent research indicates that vitamin E forms are differentially metabolized to various carboxychromanols. Some these vitamin E metabolites have been shown to exhibit strong anti-inflammatory and anticancer effects, yet little is known about their bioavailability. Without this knowledge, it is impossible to assess the role of vitamin E metabolism in biological functions of vitamin E forms and their protective effects on chronic diseases. While αT and γT appear to improved gut health, the underlying mechanisms are not well understood. Furthermore, specific forms of vitamin E such as γT have been reported to have cancer-preventing effects, but their anticancer efficacy is relatively modest. For these reasons, this dissertation focused on the characterization of the pharmacokinetic formation of vitamin E metabolites after supplementation, and the investigation of the underlying mechanisms of the protective effect of vitamin E forms, αT and γT, on gut health, as well as anticancer efficacy of the combination of aspirin and γT on carcinogen-induced colon tumorigenesis.</p><p><br></p><p>The first project focuses on characterizing the pharmacokinetic formation of vitamin E metabolites after single dose supplementation of γ-tocopherol-rich mixed tocopherol (γTmT) and δ-tocotrienol (δTE). With our recently developed LC/MS/MS assay for quantifying vitamin E metabolites, we can simultaneously quantify the level of short-chain, long-chain, and sulfated carboxychromanols in plasma, urine, and fecal samples of supplemented animals. In this study, we investigated the pharmacokinetics including excretion of vitamin E forms and the formation of their metabolites after a single dose intragastric administration of tocopherols and tocotrienols in rats. We also measured vitamin E metabolites in the serum obtained from healthy humans after gT supplementation. In the plasma of rat, the pharmacokinetic profiles of γT and δTE are described as the following: γT, Cmax = 25.6 ± 9.1 μM, Tmax = 4 h; δTE, Cmax = 16.0 ± 2.3 μM, Tmax = 2 h. Sulfated CEHCs and sulfated 11’-COOHs were the predominant metabolites in the plasma of rat with Cmax of 0.4-0.5 μM (Tmax ~ 5-7 h) or ~0.3 μM (Tmax at 4.7 h), respectively. In 24-h urine, 2.7% of γT and 0.7% of dTE were excreted as conjugated CEHCs, the major identified urinary metabolites. In the feces, 17-45% of supplemented vitamers were excreted as un-metabolized forms and 4.9-9.2% as metabolites. The majority of metabolites excreted in feces were unconjugated carboxychromanols, among which 13’-COOHs constituted ~50% of total metabolites. Interestingly, 13’-COOHs derived from δTE were 2-fold higher than 13’-COOH from γT. Unlike rats, γ-CEHC is the predominant metabolites found in human plasma, although 11’-COOHs and 13’-COOHs (sulfated and unconjugated) were elevated by >20 folds responding to γT supplement. In this study, we found that tocopherols and tocotrienols, when taken as supplements, are mainly excreted as un-metabolized forms and long-chain carboxychromanols in feces. High fecal availability of 13’-COOHs may contribute to modulating effects on gut health.</p><p><br></p><p>The second project of my dissertation investigated the effect of vitamin E forms, αT and γT, on intestinal barrier function in a cellular model and a mouse colitis model. Inflammatory bowel diseases (IBD) are chronic idiopathic inflammatory conditions characterized by disruption of intestinal barrier integrity. Previous studies by others and us had demonstrated that vitamin E forms, αT and γT, can protect against chemical-induced colitis in animal models. However, the role of these vitamin E forms on intestinal barrier function has not been studied. Herein, we investigated the potential protective effects of vitamin E forms, αT and γT, on intestinal barrier function in a Caco-2 colon epithelial cell model and a dextran sodium sulfate (DSS)-induced colitis mouse model. In Caco-2 cells, pretreatment with 25mM αT and γT attenuated Caco-2 monolayer barrier dysfunction induced by 10 ng/mL TNF-α/IFN-γ, suggesting that these vitamin E forms protect intestinal barrier integrity in this cellular model. In male BALB/c mice, the supplementation of αT (0.05%) or γTmT (0.05%) when given 3 weeks before DSS treatment or at the same time as DSS treatment alleviated DSS-induced fecal bleeding and diarrhea symptoms in mice, and attenuated colon inflammation and colitis-associated damages. Additionally, αT and γTmT supplementation attenuated DSS-induced intestinal barrier dysfunction, as indicated by improving the level of occludin, a tight junction protein, in the colon and reducing lipopolysaccharide-binding protein (LBP) in the plasma. Furthermore, gut microbiota analysis demonstrated that αT and γTmT supplementation could modulate intestinal microbiome composition in mice with DSS treatment. DSS treatment reduced the relative abundance of Lachnospiraceae compared to healthy mice, and supplementation of αT and γT partially reversed this effect. Interestingly, the family Lachnospiraceae has been reported to decrease in IBD patients. Our study demonstrated the protective effects of vitamin E forms on intestinal barrier integrity in a cell-based model and a colitis model in mice. Furthermore, we demonstrated that these vitamin E forms caused favorable changes in the intestinal microbial population under colitis condition.</p><p><br></p><p>The third project of my dissertation evaluated the anticancer efficacy of the combination of aspirin and γT using an azoxymethane (AOM)-induced and colitis-promoted colon tumorigenesis mouse model. Extensive inflammation in the colon promotes the development of colorectal cancer (CRC). Eicosanoid production by pro-inflammatory enzymes, cyclooxygenases (COX-1 and COX-2) and 5-lipoxygenase (5-LOX) play a critical role in the initiation, progression, and invasion of CRC. Thus, nonsteroidal anti-inflammatory drugs (NSAIDs), such as aspirin, have been recommended for chemoprevention of CRC. However, long-term use of aspirin can cause many side effects, and the anticancer activity of aspirin is very modest. Previously, we have demonstrated that the combination of γT with aspirin prolonged the anti-inflammatory activity of aspirin and alleviated aspirin-associated adverse effects in a carrageenan-induced inflammation model in rats. Additionally, we found that the combination of γT and aspirin has stronger anticancer activity than aspirin or γT alone against HCT-116 human colorectal carcinoma cells. Therefore, we examined the anticancer effect of the combination of 0.025% aspirin and 0.05% γT against AOM-induced and DSS-promoted tumorigenesis in mice. In this study, we have found that the combination of aspirin and γT, but not aspirin or γT alone, suppressed colon tumorigenesis in mice, as indicated by 40% and 50% reduction in the multiplicity of total polyps (P < 0.05) and large adenomatous polyps (>2mm2, P < 0.05), respectively. More strikingly, the combination of aspirin and γT reduced the overall tumor area by 60% (P < 0.05). Noteworthy, the supplementation of γT also alleviated aspirin-induced stomach lesion and appeared to modulate intestinal microbial composition. Our study demonstrated that the combination of aspirin and γT has stronger anticancer activity than aspirin or γT alone while alleviates aspirin-associated adverse effect, suggesting that the combination of γT and aspirin is a more effective and safer chemopreventive agent for CRC than aspirin alone.</p>

The task of this thesis is to familiarize readers with the basics of the problem of ionizing radiation from its types and resources, through mechanisms of action and its effects on the human organism, to the possibility of calculating various characteristics. Close attention is paid to the use of ionizing radiation in medical approaches. There is in detail described process of radiotherapy planning and a basic properties of planning systems. Part of the thesis is also program RT-Plan processed in Matlab. The program simulates the planning system for external ionizing radiation therapy used in treatment. There is supplemented a detailed help and series of tests for the plan. It is suitable for possible use in teaching.

Jamaican males are a high-risk population for aggressive prostate cancer (PrCa) due to genetic influences, and identifying empirical data on treatments, which provide survival benefits is a prime challenge for clinicians who manage Jamaican PrCa patients. Thus, the purpose of this investigation was to elucidate treatment effects of brachytherapy and ERBT in the survival of a Jamaican PrCa cohort. Differences in survival outcomes of brachytherapy and ERBT treated Jamaican, and White U.S.-born PrCa patients with localized PrCa were compared. The mechanism of radiation programmed cell death in PrCa carcinogenesis explained in the oxidative stress theory, was the theoretical base for interpreting the research questions. A retrospective cohort design was used, and included survival analysis of secondary data from the Surveillance Epidemiology and End Results database. The sample size was 10,752 Jamaican and White U.S.-born prostate cancer patients diagnosed between 1992 and 2011. Kaplan-Meier and Cox proportional hazard regression models confirmed that brachytherapy resulted in enhanced survival benefits to the Jamaicans, HR 0.63, 95% CI [0.55, 0.73], p < .001, but ERBT did not, HR 1.6, 95% CI [1.38, 1.84] p < .001. Hence, brachytherapy may be an appropriate treatment option for Jamaican PrCa patients. Clinicians and health care planners can utilize the results for policy decisions aimed at increasing access to brachytherapy treatments to Jamaicans. Improving access to efficient PrCa treatments could reduce the morbidity and mortality rates of PrCa among Jamaicans, decrease years of potential life lost from PrCa, and enhance the life expectancy of the Jamaican male population.