Despite recent advances, radiopharmaceuticals to detect and characterize bacterial infections have a number of limitations. Many of the clinically approved radiopharmaceutical agents are not specific to bacterial infections and accumulate at lesions of inflammation. Hence, new approaches are necessary to detect bacteria with high specificity and selectivity. A library of desferrioxamine B (DFO) derivatives were prepared to create radiolabeled siderophores to create a bacteria-specific imaging probe by exploiting the mechanism bacteria use to scavenge iron, which plays a key role in bacterial growth and biofilm formation Compounds were synthesized using two convenient carbamate forming strategies in 30% to 92% yield. The cold and radioactive gallium (67Ga) complexes were prepared and characterized and their uptake by S. aureus bacteria were assessed in vitro and in vivo. In vivo studies revealed that 67GaDFOethoxycarbamate had uptake comparable to GaDFO that was blockable, showing the compound was actively taken up via the siderophore pathway. In vivo studies in a mouse model resulted in a good infected to non-infected thigh ratio (11:1) and non-specific uptake by the GI tract. Bioorthogonal chemistry was also explored as an approach for imaging infection using trans-cyclooctene (TCO) functionalized vancomycin and a tetrazine functionalized 67GaDFO (67GaDFO-Tz) complex.2,3 In vitro results revealed that allowing vancomycin-TCO to bind S. aureus prior to the addition of 67GaDFO-Tz (pretargeting) showed higher (63%) uptake than with a conjugate formed prior to incubation with the bacteria (direct targeting, 28%). For the bioorthogonal approach, the distribution of the 67GaDFO-Tz was assessed in a S. aureus infection murine model, which showed significant uptake of 67GaDFO-Tz in the GI tract 1 h post intravenous injection. However, uptake in the infected joint was evident at 71 h post infection. The data suggests targeting bacteria using TCO-labeled antibiotics and radiolabeled tetrazines is a feasible strategy, but that further optimization of the vancomycin injection dose and injection time are necessary. / Thesis / Master of Science (MSc)

OBJETIVO: A imagem molecular de transportadores de dopamina (DAT) oferecer uma informação diferencial na investigação de doenças neurodegenerativas, como a Doença de Parkinson, diante de uma sólida abordagem quantitativa. Porém, são diversos o número de diferentes métodos semiquantitativos aplicados na prática clínica, que nos quais podem produzir resultados distintos quando aplicados por diferentes avaliadores ou condições de avaliação da imagem de SPECT do corpo estriado. Logo, este estudo pode avaliar a acurácia, precisão e reprodutibilidade de diferentes métodos semiquantitativos de imagens de SPECT do corpo estriado. MATERIAIS E MÉTODOS: Foram realizadas 23 aquisições de SPECT utilizando um simulador antropomórfico estriatal preenchido com diferentes concentrações de atividade de 99mTc. A preparação deste simulador estriatal foi realizada a partir de concentrações de atividades nas cavidades de interesse (núcleo caudado, putâmen e corpo estriado) proporcionalmente maiores do que a da cavidade de referência (background). As imagens foram reconstruídas utilizando parâmetros ideais já protocolados. Cinco métodos baseados em ROIs, dedicados para a quantificação de SPECT do corpo estriado foram avaliados: ROIs desenhadas sobre as imagens de SPECT - (A) Manual, ROIs com dimensões padronizadas - (B) método Twobox e (C) método Threebox; e baseado em imagens estruturais (D) MRI e (E) CT. A acurácia de cada método aplicado foi avaliada através do coeficiente de correlação de concordância (CCC), sua precisão utilizando o coeficiente de Pearson e modelos regressão linear, assim como a reprodutibilidade pode ser investigada através de análises de variabilidade intra- e interobservadores. RESULTADOS: Tanto para as cavidades avaliadas de forma individual (Caudado e Putâmen), quanto para o corpo estriado como um todo, todos os métodos aplicados apresentaram um aumento no CCC dos índices quantificados diante de uma diminuição dos valores nominais de preenchimento. Os métodos D e E apresentaram os máximos valores de CCC na avaliação do núcleo caudado _ 0,89 baixo MRI CA CCC ? e _ 0,84 baixo CT CA CCC ? ) e putâmen ( _ 0,86 baixo MRI PU CCC ? e _ 0,82 baixo CT PU CCC ? ). Entretanto, na avaliação do corpo estriado, o método B apresentou a máxima acurácia dentre os cinco aplicados ( _ 0,95 baixo TWOBOX ST CCC ? ). A significante correlação entre os métodos foi evidenciada por um elevado coeficiente de correlação (r > 0.8). Na avaliação da reprodutibilidade intra e interobservadores uma grande variabilidade foi observada na aplicação do método A, principalmente quando aplicada na semiquantificação de baixas concentrações de atividade. Conclusão: Os cinco métodos semiquantitativos de SPECT do corpo estriado, demonstraram ser eficientes na realização de leituras proporcionais dos índices de BPI mesmo quando aplicados a imagens de diferentes concentrações de atividade. Porém, diante de investigações que necessitem de um processo de quantificação mais acurado e visando avaliar putâmen e núcleo caudado de forma isolada, os métodos estruturais (MRI e CT), demonstraram uma crescente eficiência em representar acurados parâmetros semiquantitativos diante da diminuição dos índices nominais de preenchimento. Na investigação de todo o corpo estriado e carente de qualquer informação estrutural, o método TwoBox passa a ser recomendado devido sua melhor performance diante todos os métodos avaliados. / AIM: The molecular image of dopamine transporters (DAT) gives differential information in research of neurodegenerative diseases, such as Parkinson\'s, when properly approached quantitatively. Yet, each method used in clinical routine may give, or not, different results when the quantifications are applied in images of several activity levels. Hence, this study assessed the accuracy, precision and reproducibility of striatum SPECT images semi-quantification methods, based in ROIs. METHODOLOGY: Twenty-three SPECT images were acquisitions of anthropomorphic striatal phantom filled with different activity concentrations of 99mTc. For each acquisition performed, the specific chambers (caudate and putamen chambers) to large chamber (simulating nonspecific background activity) was filled with solutions activity of different specific to nonspecific ratios (10, 8, 6, 4 and 2 to 1). The images were reconstructed by iterative algorithm, corrected to attenuation effects and the extracted values were analyzed by the specific binding ratio (SBR). Five semi-quantification methods for striatum SPECT, using ROIs was assessment: (A) draw freehand ROIs on SPECT image (manual); standard size ROIs: (B) TwoBox and (C) ThreeBox Methods and VOIs using structural images: (D) MRI and (E) CT. Accuracy of methods applied was assessed by concordance correlation coefficient (CCC) and precision by Pearson\'s coefficient and linear regression. RESUlTS: The SBR quantified both to individual specific chambers and striatal chamber analyzed to all methods applied resulted in a CCC increase with decrease of the nominal values used. For lower SBR values, the D and E methods evidenced the maximum values of CCC in assessment of caudate (CCCMRI_CA = 0.89 e CCCCT_CA = 0.84) and putamen (CCCMRI_PU = 0.86 e CCCCT_PU = 0.82). However, striatal assessments the B method highlights a maximum accuracy between all methods applied (CCCTWOBOX_ST = 0.95), for low values of SBR. A high Pearson\'s coefficient was found in the correlation between the all methods, report thereby a good precision between them (r > 0.8). The high ICC and variability values, showed a high reprodutibility intra- and interobserver for B, C and D methods, white the A and E methods presented a high variability between the raters. CONCLUSION: The five semi-quantification methods of striatum SPECT reported a high precision even when applied in images with activity solutions different. Therefore, to research much accurate and need to assessment just caudate or putamen individually the structural methods - MRI and CT - demonstrated progressive improvement in its quantification to reduction nominal fill index. To assessment striatal chamber and in the absence of structural information, the TwoBox method is advisable due to its excellent agreement with all nominal values when compared to the various semi-quantitative methods investigated. Keywords:

Introdução: A presença de cálcio nas artérias coronárias identifica a existência de aterosclerose coronariana, mesmo em fases precoces. Por outro lado, a decisão quanto à melhor forma de tratamento dessa entidade baseia-se no uso de exames funcionais, em especial, a cintilografia de perfusão do miocárdio (CPM). Existem dúvidas quanto à correlação desses dois exames, o que poderia ser, ao menos em parte, explicada pelo fato dos testes avaliarem fases distintas dessa entidade. Este estudo pretende avaliar o uso de uma abordagem anatômica e funcional combinada por meio da (CPM) e tomografia computadorizada coronária com multidetectores (TCMD) na determinação do escore de cálcio (CACS) e da presença de estenoses e isquemia e examinar a associação de seus resultados entre si e com outras variáveis demográficas, clínicas e funcionais em uma população brasileira com doença arterial coronária (DAC) suspeita ou conhecida. Métodos: foram analisados, retrospectivamente, 413 pacientes que se submeteram à CPM e TCMD por indicação clínica do médico assistente, durante o ano de 2009, com intervalo menor de 90 dias, entre os dois estudos. CACS foi definido automaticamente pelo software dedicado e o escore de Agatston foi calculado de forma semiquatitativa. Na CPM e na TCMD, os resultados foram obtidos por quantificação visual semiobjetiva. A correlação dos resultados dos exames foi analisada e foram obtidos dados da evolução tardia, baseados no contato com o médico assistente ou pela análise dos registros hospitalares. Procurou-se definir os preditores da ocorrência de eventos cardiovasculares adversos na evolução. Resultados: Foram selecionados 303 pacientes (73,3% homens, média de idade de 55,8 ± 10,6 anos, intervalo: 32-86 anos). Destes, 73,3% apresentaram estudos de perfusão normal, 71,6% tinham cálcio coronário e 45,2% tinha perfusão normal e TCDM com cálcio. No grupo sem suspeita de DAC (177), houve associação entre os resultados de TCMD e SPECT com: diabetes (DM) (p=0,045), hipertensão (HAS) (p=0,032), dislipidemia (p=0,030) testes funcionais, resultado do teste (p=0,022), percentil escore de cálcio (p<0,001) e CACS (p<0,001). Nos pacientes sem DAC, houve associação com defeitos de perfusão e TCMD com cálcio que mostraram em casos de DM (30,0%) e CACS igual ou acima do percentil 75 (60,0%) e com valores acima de 400 (40,0%). O grupo com TCMD e perfusão normal apresentou maior percentual de ausência de HAS (78,6%). De outra forma, o grupo com perfusão normal e TCMD com cálcio mostrou mais pacientes com dislipidemia (42,5%) e distribuição dos percentis de CACS e CACS, semelhantes ao grupo com defeitos de perfusão e TCMD com cálcio. No grupo dos pacientes com DAC conhecida, também houve associação entre CACS e a extensão da aterosclerose coronária. Não houve associações entre os resultados da cintilografia e TCMD e outras variáveis. No subgrupo de 128 pacientes, em que se conseguiu seguimento tardio (média de 824,5 dias; DP de 385,9), aconteceram dois óbitos (1,6%), nenhum de causa cardiovascular. O evento mais frequente foi cinecoronariografia (CAT) (21,1%), seguida da angioplastia (ATC) (9,4%) e revascularizações cirúrgicas (3,1%). Não houve infarto do miocárdio (IM). De forma similar, não foram observados determinantes significativos da evolução tardia desses pacientes. Conclusões: Embora o CAC tenha sido tão frequente, como perfusão normal no SPECT, menos da metade daqueles com SPECT normal podem apresentar cálcio coronário à TCMD. Esta combinação de resultados (SPECT normal e cálcio) tinha associação com dislipidemia. Além disso, DM, CACS > 400 e percentil igual ou superior a 75% foram associados com SPECT anormal e cálcio na MDCT. A MDTC combinada ao SPECT foi capaz de detectar isquemia em pacientes com estenose coronariana conhecida, mas não havia associações entre o grau de estenose ou a presença, ou ausência de isquemia e sua extensão, com os resultados combinados. Houve uma baixa taxa de eventos no subgrupo de acompanhamento. / Background: The calcium in coronary arteries identifies the presence of coronary atherosclerosis, even in early stages. On the other hand, the decision about the best form of treatment this entity is based on the use of functional tests, in particular the myocardial perfusion scintigraphy (SPECT). There are doubts about the correlation between the results of these two exams, what could be, at least in part explained by the fact of these tests assess distinct phases of this entity. This study aims to evaluate the use of a combined functional and anatomical approach through SPECT and coronary computed tomography with multi-detectors (MDCT) in determining the calcium score (CACS),presence of stenosis or ischemia and examine the combined results with other demographic, clinical and functional variables, in a Brazilian population with suspected or known coronary artery disease (CAD). Methods: we retrospectively analyzed 413 patients who underwent to SPECT and MDCT by their physician indications during the year 2009, with less than 90 days interval between the two studies. CACS was automatically defined by dedicated software and the Agatston score was semi-automatically calculated. SPECT and MDCT results were evaluated by semi-objective visual quantification. The correlations of both tests results were analyzed and follow-up data were obtained through contact with assistant physician or analysis of hospital records. Results: We included 303 patients with suspect or known DAC who underwent to both, SPECT and MDCT (75.9 % men, mean age 55.8 ± 10.6 years; range:32-86 years); it was observed that 73,3% had normal perfusion studies, CAC was present 71,6% patients and 48.2% had normal perfusion and MDCT with calcium. In 177 patients without DAC there were association between MDCT and SPECT results and following variables: diabetes (DM) (p=0.045), hypertension (HAS) (p=0.032), dyslipidemia (p=0.030), functional test results (p=0.022), percentile of calcium score (p<0.001) and CACS (p<0.001). Patients without DAC, with perfusion defects and calcium showed more occurrences of DM (30.0%) and CACS equal to or above the 75 percentile (60.0%) and with values above 400 (40.0%). The group with normal MDCT and perfusion had higher percentage of absence of HAS (78.6%). The group with normal perfusion and MDCT with calcium had the highest percentage of patients with dyslipidemia (42.5%) and distribution of the calcium score percentiles and CACS were similar to the group with perfusion defects and MDCT with calcium. In the group with known CAD there was association between DM, extent of coronary disease and CACS. There were no associations between the results of SPECT and MDCT and other variables. In the subgroup of 128 patients with follow-up, the average time was 824.5 days (SD 385.9), there were 2 deaths (1.6%) confirmed, however none of cardiovascular cause. Coronary angiography (CATH) (21.1%) was the most frequent event, followed by percutaneous angioplasty (PTCA) (9.4%). Surgical revascularizations were much less frequent (3.1%). There was no myocardial infarction (MI). Similarly, there were no markers of long term prognosis in this sample. Conclusions: Although CAC was as frequent as normal perfusion in SPECT, less than half with normal SPECT may have calcium on MDCT. This results combination (normal SPECT and calcium) had association with dyslipidemia. Also, DM, CACS >400 and percentile equal or above 75% were associate with abnormal SPECT and calcium on MDCT. MDCT combined to SPECT was able to detect ischemia in patients with known coronary stenosis, but there were no associations between the degree of stenosis, or the presence or absence of ischemia and its extent with the combined results. There was a low event rate in the follow-up subgroup.

OBJETIVO: A imagem molecular de transportadores de dopamina (DAT) oferecer uma informação diferencial na investigação de doenças neurodegenerativas, como a Doença de Parkinson, diante de uma sólida abordagem quantitativa. Porém, são diversos o número de diferentes métodos semiquantitativos aplicados na prática clínica, que nos quais podem produzir resultados distintos quando aplicados por diferentes avaliadores ou condições de avaliação da imagem de SPECT do corpo estriado. Logo, este estudo pode avaliar a acurácia, precisão e reprodutibilidade de diferentes métodos semiquantitativos de imagens de SPECT do corpo estriado. MATERIAIS E MÉTODOS: Foram realizadas 23 aquisições de SPECT utilizando um simulador antropomórfico estriatal preenchido com diferentes concentrações de atividade de 99mTc. A preparação deste simulador estriatal foi realizada a partir de concentrações de atividades nas cavidades de interesse (núcleo caudado, putâmen e corpo estriado) proporcionalmente maiores do que a da cavidade de referência (background). As imagens foram reconstruídas utilizando parâmetros ideais já protocolados. Cinco métodos baseados em ROIs, dedicados para a quantificação de SPECT do corpo estriado foram avaliados: ROIs desenhadas sobre as imagens de SPECT - (A) Manual, ROIs com dimensões padronizadas - (B) método Twobox e (C) método Threebox; e baseado em imagens estruturais (D) MRI e (E) CT. A acurácia de cada método aplicado foi avaliada através do coeficiente de correlação de concordância (CCC), sua precisão utilizando o coeficiente de Pearson e modelos regressão linear, assim como a reprodutibilidade pode ser investigada através de análises de variabilidade intra- e interobservadores. RESULTADOS: Tanto para as cavidades avaliadas de forma individual (Caudado e Putâmen), quanto para o corpo estriado como um todo, todos os métodos aplicados apresentaram um aumento no CCC dos índices quantificados diante de uma diminuição dos valores nominais de preenchimento. Os métodos D e E apresentaram os máximos valores de CCC na avaliação do núcleo caudado _ 0,89 baixo MRI CA CCC ? e _ 0,84 baixo CT CA CCC ? ) e putâmen ( _ 0,86 baixo MRI PU CCC ? e _ 0,82 baixo CT PU CCC ? ). Entretanto, na avaliação do corpo estriado, o método B apresentou a máxima acurácia dentre os cinco aplicados ( _ 0,95 baixo TWOBOX ST CCC ? ). A significante correlação entre os métodos foi evidenciada por um elevado coeficiente de correlação (r > 0.8). Na avaliação da reprodutibilidade intra e interobservadores uma grande variabilidade foi observada na aplicação do método A, principalmente quando aplicada na semiquantificação de baixas concentrações de atividade. Conclusão: Os cinco métodos semiquantitativos de SPECT do corpo estriado, demonstraram ser eficientes na realização de leituras proporcionais dos índices de BPI mesmo quando aplicados a imagens de diferentes concentrações de atividade. Porém, diante de investigações que necessitem de um processo de quantificação mais acurado e visando avaliar putâmen e núcleo caudado de forma isolada, os métodos estruturais (MRI e CT), demonstraram uma crescente eficiência em representar acurados parâmetros semiquantitativos diante da diminuição dos índices nominais de preenchimento. Na investigação de todo o corpo estriado e carente de qualquer informação estrutural, o método TwoBox passa a ser recomendado devido sua melhor performance diante todos os métodos avaliados. / AIM: The molecular image of dopamine transporters (DAT) gives differential information in research of neurodegenerative diseases, such as Parkinson\'s, when properly approached quantitatively. Yet, each method used in clinical routine may give, or not, different results when the quantifications are applied in images of several activity levels. Hence, this study assessed the accuracy, precision and reproducibility of striatum SPECT images semi-quantification methods, based in ROIs. METHODOLOGY: Twenty-three SPECT images were acquisitions of anthropomorphic striatal phantom filled with different activity concentrations of 99mTc. For each acquisition performed, the specific chambers (caudate and putamen chambers) to large chamber (simulating nonspecific background activity) was filled with solutions activity of different specific to nonspecific ratios (10, 8, 6, 4 and 2 to 1). The images were reconstructed by iterative algorithm, corrected to attenuation effects and the extracted values were analyzed by the specific binding ratio (SBR). Five semi-quantification methods for striatum SPECT, using ROIs was assessment: (A) draw freehand ROIs on SPECT image (manual); standard size ROIs: (B) TwoBox and (C) ThreeBox Methods and VOIs using structural images: (D) MRI and (E) CT. Accuracy of methods applied was assessed by concordance correlation coefficient (CCC) and precision by Pearson\'s coefficient and linear regression. RESUlTS: The SBR quantified both to individual specific chambers and striatal chamber analyzed to all methods applied resulted in a CCC increase with decrease of the nominal values used. For lower SBR values, the D and E methods evidenced the maximum values of CCC in assessment of caudate (CCCMRI_CA = 0.89 e CCCCT_CA = 0.84) and putamen (CCCMRI_PU = 0.86 e CCCCT_PU = 0.82). However, striatal assessments the B method highlights a maximum accuracy between all methods applied (CCCTWOBOX_ST = 0.95), for low values of SBR. A high Pearson\'s coefficient was found in the correlation between the all methods, report thereby a good precision between them (r > 0.8). The high ICC and variability values, showed a high reprodutibility intra- and interobserver for B, C and D methods, white the A and E methods presented a high variability between the raters. CONCLUSION: The five semi-quantification methods of striatum SPECT reported a high precision even when applied in images with activity solutions different. Therefore, to research much accurate and need to assessment just caudate or putamen individually the structural methods - MRI and CT - demonstrated progressive improvement in its quantification to reduction nominal fill index. To assessment striatal chamber and in the absence of structural information, the TwoBox method is advisable due to its excellent agreement with all nominal values when compared to the various semi-quantitative methods investigated. Keywords:

Os distúrbios da articulação temporomandibulares (DTM) apresentam um impacto negativo na qualidade de vida de seus portadores. A diversidade de sinais e sintomas apresentados pelos pacientes, como dores musculares, cefaléias (difusa ou localizada), dores na região da articulação temporomandibular (ATM), limitações ou excessos de abertura da boca e ruídos articulares dificultam o diagnóstico clínico. Para isso, há necessidade de uma investigação clínica minuciosa e de exames complementares. Os exames radiográficos convencionais e tomografias mostram a anatomia da região sem demonstrar alterações funcionais locais. A técnica de imagens tomográficas em Medicina Nuclear, SPECT (Single Photon Emission Computed Tomography) com 99m Tc-MDP (radiofármaco composto de Tecnécio 99 metaestável e Metileno Difosfonato), demonstra o metabolismo da região, e associada à fusão de imagens co-registradas com a tomografia computadorizada (CT), permite a localização anatômica das alterações ósseas funcionais existentes nas estruturas envolvidas. É um exame com baixa dose de radiação que possibilita a observação conjunta estrutural anatômica e metabólica. Neste trabalho avaliamos, retrospectivamente, exames SPECT/CT de 33 pacientes realizados no Setor de Medicina Nuclear do Departamento de Imagem do Centro de Tratamento e Pesquisa - Hospital do Câncer A. C. Camargo, que apresentavam sinais e sintomas de provável DTM. Os resultados dos exames foram relacionados aos dados existentes nos prontuários dos pacientes, como sexo, idade, anamnese e exame clínico. Por meio de análise descritiva, utilizando as freqüências e porcentagens das variáveis de interesse, pudemos avaliar a confiabilidade do exame. Concluímos que o SPECT/CT, quando aplicado em pacientes portadores de sinais e sintomas de DTM, apresentou alta sensibilidade (100%), especificidade (90,9%) e acurácia (96,9%), demonstrando que é um método de diagnóstico por imagem de relevância, útil e de grande valia, podendo ser um dos métodos de escolha indicados para diagnóstico de DTM. / Temporomandibular joint dysfunction play a negative role on patient life quality standard. A diversity of signs and symptoms such us muscular pain, headaches (focal or diffuse), temporomandibular joint localized pain, limitation or excessive mandibular opening and joint noise may lead to difficulties of correct diagnosis. Clinical investigation and complementary exams are necessary in these cases. Conventional radiographic and tomographic exams exactilly indicates local anatomy but do not demonstrate functional metabolic changes. Tomographic images on Nuclear Medicine, SPECT (Single Photon Emission Computed Tomography) with 99m Tc-MDP (methylene diphosphonate) a radiopharmaceutical labeled with Technetium 99m demonstrate regional metabolism changes. The association to co-registered computed tomography (CT) and image fusion allows the anatomic placement of bone functional alteration into the involved structures. SPECT/CT is a low dose radiation procedure that makes simultaneous anatomic and metabolic observation possible. This assignment was retrospectively analyzed, SPECT/CT of 33 patients from the Nuclear Medicine Sector of the Image Department of the Research and Treatment Center of the Cancer Hospital A. C. Camargo, that presented sings and symptoms of probable temporomandibular joint dysfunction. The results of the exams were related with the patient data that were on their file like sex, age, anamnesis and clinical exam. The assessment of the reliability of the exam was evaluated by descriptive analysis using frequencies and percentages of the interest variables. We conclude that SPEC/CT showed high sensitivity (100%), specificity (90,9%) and accuracy (96,9%); demonstrating that it is an important and useful diagnostic method, that may be an exam of choice to the diagnostic features of temporomandibular joint disfunction.

Os distúrbios da articulação temporomandibulares (DTM) apresentam um impacto negativo na qualidade de vida de seus portadores. A diversidade de sinais e sintomas apresentados pelos pacientes, como dores musculares, cefaléias (difusa ou localizada), dores na região da articulação temporomandibular (ATM), limitações ou excessos de abertura da boca e ruídos articulares dificultam o diagnóstico clínico. Para isso, há necessidade de uma investigação clínica minuciosa e de exames complementares. Os exames radiográficos convencionais e tomografias mostram a anatomia da região sem demonstrar alterações funcionais locais. A técnica de imagens tomográficas em Medicina Nuclear, SPECT (Single Photon Emission Computed Tomography) com 99m Tc-MDP (radiofármaco composto de Tecnécio 99 metaestável e Metileno Difosfonato), demonstra o metabolismo da região, e associada à fusão de imagens co-registradas com a tomografia computadorizada (CT), permite a localização anatômica das alterações ósseas funcionais existentes nas estruturas envolvidas. É um exame com baixa dose de radiação que possibilita a observação conjunta estrutural anatômica e metabólica. Neste trabalho avaliamos, retrospectivamente, exames SPECT/CT de 33 pacientes realizados no Setor de Medicina Nuclear do Departamento de Imagem do Centro de Tratamento e Pesquisa - Hospital do Câncer A. C. Camargo, que apresentavam sinais e sintomas de provável DTM. Os resultados dos exames foram relacionados aos dados existentes nos prontuários dos pacientes, como sexo, idade, anamnese e exame clínico. Por meio de análise descritiva, utilizando as freqüências e porcentagens das variáveis de interesse, pudemos avaliar a confiabilidade do exame. Concluímos que o SPECT/CT, quando aplicado em pacientes portadores de sinais e sintomas de DTM, apresentou alta sensibilidade (100%), especificidade (90,9%) e acurácia (96,9%), demonstrando que é um método de diagnóstico por imagem de relevância, útil e de grande valia, podendo ser um dos métodos de escolha indicados para diagnóstico de DTM. / Temporomandibular joint dysfunction play a negative role on patient life quality standard. A diversity of signs and symptoms such us muscular pain, headaches (focal or diffuse), temporomandibular joint localized pain, limitation or excessive mandibular opening and joint noise may lead to difficulties of correct diagnosis. Clinical investigation and complementary exams are necessary in these cases. Conventional radiographic and tomographic exams exactilly indicates local anatomy but do not demonstrate functional metabolic changes. Tomographic images on Nuclear Medicine, SPECT (Single Photon Emission Computed Tomography) with 99m Tc-MDP (methylene diphosphonate) a radiopharmaceutical labeled with Technetium 99m demonstrate regional metabolism changes. The association to co-registered computed tomography (CT) and image fusion allows the anatomic placement of bone functional alteration into the involved structures. SPECT/CT is a low dose radiation procedure that makes simultaneous anatomic and metabolic observation possible. This assignment was retrospectively analyzed, SPECT/CT of 33 patients from the Nuclear Medicine Sector of the Image Department of the Research and Treatment Center of the Cancer Hospital A. C. Camargo, that presented sings and symptoms of probable temporomandibular joint dysfunction. The results of the exams were related with the patient data that were on their file like sex, age, anamnesis and clinical exam. The assessment of the reliability of the exam was evaluated by descriptive analysis using frequencies and percentages of the interest variables. We conclude that SPEC/CT showed high sensitivity (100%), specificity (90,9%) and accuracy (96,9%); demonstrating that it is an important and useful diagnostic method, that may be an exam of choice to the diagnostic features of temporomandibular joint disfunction.

Actuellement, les injections systémiques de médicaments atteignent faiblement les sites tumoraux et de fortes doses sont alors administrées provoquant des effets secondaires parfois lourds. Les possibilités offertes par les applications en médecine des nanoparticules permettent de nouvelles stratégies pour vectoriser des substances actives dans les cellules malades. Ces travaux de thèse portent sur le cancer de la prostate qui est le deuxième cancer le plus diagnostiqué et la cinquième cause de décès chez les hommes dans le monde.Les nanotubes de titanate (TiONts) sont synthétisés par voie hydrothermale et présentent une longueur moyenne de 170 nm, un diamètre extérieur de 10 nm et une cavité interne accessible de 4 nm. Leur morphologie tubulaire permet aux TiONts d’être internalisés plus facilement dans les cellules, sans induire de cytotoxicité, tout en créant un effet radiosensibilisant.Deux nanohybrides ont été développés dans cette thèse, pour lutter contre le cancer de la prostate par injection intratumorale (IT) et une attention particulière a été portée sur leur élaboration. Ces nouveaux nanomédicaments ont été pleinement caractérisés par différentes techniques (MET, ATG, potentiel zêta, XPS, spectroscopies UV visible, IR et Raman).La première approche consiste à combiner les TiONts avec un agent thérapeutique (le docétaxel, DTX), largement utilisé pour inhiber les tumeurs de prostate et un agent chélatant (le DOTA, radiomarqué avec l’111In) pour suivre la biodistribution des tubes par SPECT/CT. La surface des TiONts a été préalablement fonctionnalisée par l’APTES et le poly(éthylène) glycol (PEG3000) pour rendre les TiONts stables et biocompatibles. Afin d’évaluer l’efficacité de ce nanohybride, des tests in vitro ont montré que l’association entre les TiONts et le DTX permettait de maintenir une activité cytotoxique sur des lignées cellulaires de prostate (cellules 22Rv1 et PC 3) alors que les TiONts sans le DTX n’étaient pas toxiques. Les études in vivo ont montré, sur des souris Swiss nude mâles, que plus de 70% des nanovecteurs étaient retenus dans la tumeur, après injection IT, après 7 jours. De plus, un retard de croissance tumorale pour les souris ayant reçu le nanohybride avec la radiothérapie (RT) est observé, par rapport aux souris ayant reçues seulement le DTX. Après cette étude, d’autres molécules organiques ont été greffées avec succès à la surface des TiONts pour améliorer la stabilité colloïdale et la biocompatibilité des nanotubes : AHAMTES, catéchols (LDOPA, DHCA et NDOPA), phosphonates (PHA, ALD et un polymère hétérobifonctionnel de type phosphonate : (HO)2 (O)P-PEG NH2). De plus, le greffage de différentes longueurs de chaîne de PEG a été évalué par deux voies de synthèses. Le greffage de ces PEG en milieu organique (PyBOP) s’est avéré très prometteur pour améliorer leur taux de greffage et leur stabilité colloïdale.Dans une seconde approche, pour accroître l’effet radiosensibilisant, des nanoparticules d’or (AuNPs), elles-mêmes modifiées par le DTDTPA, ont été couplées avec les TiONts en présence ou non de DTX. Cette nouvelle combinaison a pour objectif le maintien des AuNPs, par les TiONts, dans la tumeur afin d’améliorer l’effet de la RT. Grâce aux AuNPs modifiées par le DTDTPA, le nanohybride est également détectable par imagerie X et par SPECT/CT. Les résultats in vitro ont démontré l’activité cytotoxique de l’édifice final. Des études de biodistribution et de croissance tumorale ont également été réalisées sur des tumeurs PC-3 xénogreffées sur des souris.Ces TiONts fonctionnalisés apparaissent comme un nouvel outil polyvalent dans le domaine médical, notamment pour lutter contre le cancer de la prostate. / Currently, the systemic injections of drugs reach very weakly tumor sites and large doses are thus administered causing adverse side effects. The new implementations of nanoparticles in the medical field offer new strategies to vectorize an active substance in diseased cells. This work is focused on the prostate cancer, which is the second most frequently diagnosed cancer and the fifth leading cause of cancer death in men worldwide.Titanate nanotubes (TiONts) are synthetized by a hydrothermal process and have average dimensions of about 170 nm in length, 10 nm in outer diameter and also have an internal cavity of 4 nm in diameter. Their needle-shaped morphology allows them to be internalized more easily into cells without inducing cytotoxicity while providing a radiosensitization effect.In the present manuscript are described two TiONts-based nanohybrids which were developed with a view to fight against prostate cancer by intratumoral (IT) injection and a particular attention was paid on their elaboration. These new nanomedicines were extensively characterized by different techniques (TEM, TGA, ζ potential, XPS, UV visible, IR and Raman spectroscopies).The first approach that has been developed consists in combining TiONts with a therapeutic agent (docetaxel, DTX), widely used for the treatment of prostate cancer, and a chelating agent (DOTA) allowing the radiolabeling with 111In radionuclide to monitor TiONts biodistribution by SPECT/CT. The surface of TiONts was beforehand coated with a siloxane (APTES) and linked to a heterobifunctional polymer (PEG3000) to create well-dispersed and biocompatible TiONts. In vitro tests demonstrated that the association between TiONts and DTX had cytotoxic activity against prostate cancer cell lines (22Rv1 and PC-3 cells) whereas TiONts without DTX did not. The results of in vivo SPECT/CT imaging are also presented as well as first irradiation tests in Swiss nude mice after IT injection on PC-3 tumors. Biological tests showed that more than 70% of TiONts nanovectors were retained within the tumor for at least 7 days. In addition, tumor growth of mice receiving nanohybrids with radiotherapy was significantly slower than that of mice receiving free DTX. After this first study, other organic molecules were successfully grafted to the surface of TiONts to improve colloidal stability and biocompatibility of nanotubes: AHAMTES, catechols (LDOPA, DHCA and NDOPA) and phosphonates (PHA, ALD and a phosphonate heterobifunctional polymer-based: (HO)2 (O)P PEG NH2). Moreover, the influence of different PEG lengths has been considered on the nanomedicine efficacy by two different pathways. The grafting of these PEG in an organic medium (PyBOP) was very promising to improve their graft ratio and their colloidal stability.In a second approach and in order to improve the radiosensitizing effect, DTDTPA-modified gold nanoparticles (AuNPs) were coupled with TiONts in the presence of DTX. This novel combination aims at retaining these AuNPs into the tumor via the TiONts to enhance the radiotherapeutic effect. The nanohybrid was also detectable by X-ray and SPECT/CT imaging through AuNPs-DTDTPA. Preliminary in vitro results showed once again that our final nanohybrid had a satisfactory cytotoxic activity. Biodistribution and tumor growth studies were also realized on PC-3 xenografted tumors on mice.These functionalized-TiONts could thus become a new tool in the field of biomedicine to fight against prostate cancer and appear as versatile nanovectors.

Introdução: A presença de cálcio nas artérias coronárias identifica a existência de aterosclerose coronariana, mesmo em fases precoces. Por outro lado, a decisão quanto à melhor forma de tratamento dessa entidade baseia-se no uso de exames funcionais, em especial, a cintilografia de perfusão do miocárdio (CPM). Existem dúvidas quanto à correlação desses dois exames, o que poderia ser, ao menos em parte, explicada pelo fato dos testes avaliarem fases distintas dessa entidade. Este estudo pretende avaliar o uso de uma abordagem anatômica e funcional combinada por meio da (CPM) e tomografia computadorizada coronária com multidetectores (TCMD) na determinação do escore de cálcio (CACS) e da presença de estenoses e isquemia e examinar a associação de seus resultados entre si e com outras variáveis demográficas, clínicas e funcionais em uma população brasileira com doença arterial coronária (DAC) suspeita ou conhecida. Métodos: foram analisados, retrospectivamente, 413 pacientes que se submeteram à CPM e TCMD por indicação clínica do médico assistente, durante o ano de 2009, com intervalo menor de 90 dias, entre os dois estudos. CACS foi definido automaticamente pelo software dedicado e o escore de Agatston foi calculado de forma semiquatitativa. Na CPM e na TCMD, os resultados foram obtidos por quantificação visual semiobjetiva. A correlação dos resultados dos exames foi analisada e foram obtidos dados da evolução tardia, baseados no contato com o médico assistente ou pela análise dos registros hospitalares. Procurou-se definir os preditores da ocorrência de eventos cardiovasculares adversos na evolução. Resultados: Foram selecionados 303 pacientes (73,3% homens, média de idade de 55,8 ± 10,6 anos, intervalo: 32-86 anos). Destes, 73,3% apresentaram estudos de perfusão normal, 71,6% tinham cálcio coronário e 45,2% tinha perfusão normal e TCDM com cálcio. No grupo sem suspeita de DAC (177), houve associação entre os resultados de TCMD e SPECT com: diabetes (DM) (p=0,045), hipertensão (HAS) (p=0,032), dislipidemia (p=0,030) testes funcionais, resultado do teste (p=0,022), percentil escore de cálcio (p<0,001) e CACS (p<0,001). Nos pacientes sem DAC, houve associação com defeitos de perfusão e TCMD com cálcio que mostraram em casos de DM (30,0%) e CACS igual ou acima do percentil 75 (60,0%) e com valores acima de 400 (40,0%). O grupo com TCMD e perfusão normal apresentou maior percentual de ausência de HAS (78,6%). De outra forma, o grupo com perfusão normal e TCMD com cálcio mostrou mais pacientes com dislipidemia (42,5%) e distribuição dos percentis de CACS e CACS, semelhantes ao grupo com defeitos de perfusão e TCMD com cálcio. No grupo dos pacientes com DAC conhecida, também houve associação entre CACS e a extensão da aterosclerose coronária. Não houve associações entre os resultados da cintilografia e TCMD e outras variáveis. No subgrupo de 128 pacientes, em que se conseguiu seguimento tardio (média de 824,5 dias; DP de 385,9), aconteceram dois óbitos (1,6%), nenhum de causa cardiovascular. O evento mais frequente foi cinecoronariografia (CAT) (21,1%), seguida da angioplastia (ATC) (9,4%) e revascularizações cirúrgicas (3,1%). Não houve infarto do miocárdio (IM). De forma similar, não foram observados determinantes significativos da evolução tardia desses pacientes. Conclusões: Embora o CAC tenha sido tão frequente, como perfusão normal no SPECT, menos da metade daqueles com SPECT normal podem apresentar cálcio coronário à TCMD. Esta combinação de resultados (SPECT normal e cálcio) tinha associação com dislipidemia. Além disso, DM, CACS > 400 e percentil igual ou superior a 75% foram associados com SPECT anormal e cálcio na MDCT. A MDTC combinada ao SPECT foi capaz de detectar isquemia em pacientes com estenose coronariana conhecida, mas não havia associações entre o grau de estenose ou a presença, ou ausência de isquemia e sua extensão, com os resultados combinados. Houve uma baixa taxa de eventos no subgrupo de acompanhamento. / Background: The calcium in coronary arteries identifies the presence of coronary atherosclerosis, even in early stages. On the other hand, the decision about the best form of treatment this entity is based on the use of functional tests, in particular the myocardial perfusion scintigraphy (SPECT). There are doubts about the correlation between the results of these two exams, what could be, at least in part explained by the fact of these tests assess distinct phases of this entity. This study aims to evaluate the use of a combined functional and anatomical approach through SPECT and coronary computed tomography with multi-detectors (MDCT) in determining the calcium score (CACS),presence of stenosis or ischemia and examine the combined results with other demographic, clinical and functional variables, in a Brazilian population with suspected or known coronary artery disease (CAD). Methods: we retrospectively analyzed 413 patients who underwent to SPECT and MDCT by their physician indications during the year 2009, with less than 90 days interval between the two studies. CACS was automatically defined by dedicated software and the Agatston score was semi-automatically calculated. SPECT and MDCT results were evaluated by semi-objective visual quantification. The correlations of both tests results were analyzed and follow-up data were obtained through contact with assistant physician or analysis of hospital records. Results: We included 303 patients with suspect or known DAC who underwent to both, SPECT and MDCT (75.9 % men, mean age 55.8 ± 10.6 years; range:32-86 years); it was observed that 73,3% had normal perfusion studies, CAC was present 71,6% patients and 48.2% had normal perfusion and MDCT with calcium. In 177 patients without DAC there were association between MDCT and SPECT results and following variables: diabetes (DM) (p=0.045), hypertension (HAS) (p=0.032), dyslipidemia (p=0.030), functional test results (p=0.022), percentile of calcium score (p<0.001) and CACS (p<0.001). Patients without DAC, with perfusion defects and calcium showed more occurrences of DM (30.0%) and CACS equal to or above the 75 percentile (60.0%) and with values above 400 (40.0%). The group with normal MDCT and perfusion had higher percentage of absence of HAS (78.6%). The group with normal perfusion and MDCT with calcium had the highest percentage of patients with dyslipidemia (42.5%) and distribution of the calcium score percentiles and CACS were similar to the group with perfusion defects and MDCT with calcium. In the group with known CAD there was association between DM, extent of coronary disease and CACS. There were no associations between the results of SPECT and MDCT and other variables. In the subgroup of 128 patients with follow-up, the average time was 824.5 days (SD 385.9), there were 2 deaths (1.6%) confirmed, however none of cardiovascular cause. Coronary angiography (CATH) (21.1%) was the most frequent event, followed by percutaneous angioplasty (PTCA) (9.4%). Surgical revascularizations were much less frequent (3.1%). There was no myocardial infarction (MI). Similarly, there were no markers of long term prognosis in this sample. Conclusions: Although CAC was as frequent as normal perfusion in SPECT, less than half with normal SPECT may have calcium on MDCT. This results combination (normal SPECT and calcium) had association with dyslipidemia. Also, DM, CACS >400 and percentile equal or above 75% were associate with abnormal SPECT and calcium on MDCT. MDCT combined to SPECT was able to detect ischemia in patients with known coronary stenosis, but there were no associations between the degree of stenosis, or the presence or absence of ischemia and its extent with the combined results. There was a low event rate in the follow-up subgroup.

Introdução: A doença de Wilson (DW) é uma doença genética rara, sistêmica, causada por déficit no metabolismo do cobre, levando a acúmulo em diversos órgãos, incluindo o cérebro, especialmente os núcleos da base. Apesar de distonia e disartria serem as alterações neurológicas mais comuns, alterações cognitivas, incluindo demência, podem surgir em casos não tratados. Objetivos: Avaliar as funções cognitivas de um grupo de pacientes com DW e correlacionar com os achados de RM e SPECT. Métodos: Todos os pacientes com DW atendidos consecutivamente em uma clínica de distúrbios do movimento, entre outubro de 2006 e agosto de 2007, foram submetidos ao Miniexame do Estado Mental (MEEM), Bateria Cognitiva Breve (BCB), teste de fluência verbal (animais, FAS e verbos), Bateria de Avaliação Frontal (BAF), STROOP, Extensão de Dígitos (ED) ordem direta e ordem inversa, Escala de Demência de Mattis (DRS), Teste de Seleção de Cartas de Wisconsin (WCST), Hooper e Blocos do WAIS. Depois dos testes, todos os pacientes realizaram RM e SPECT. Alterações de RM (hipersinal, hiposinal e atrofia) foram quantificadas em uma escala (0-17). O SPECT foi analisado por meio de SPM. Pacientes com depressão ou com anartria foram excluídos. Os resultados foram comparados com o desempenho de um grupo de voluntários sadios. Resultados: 20 pacientes com DW (11 homens) e 20 controles (9 homens) foram avaliados. A média de idade no grupo de pacientes com DW e controles foi 30,05±7,25 anos e 32,15±5,37 anos, respectivamente. A escolaridade média foi 11,15±3,73 anos nos pacientes com DW e 10,08 ± 2,62 anos nos controles. Pacientes com DW tiveram comprometimento cognitivo nos seguintes testes: MEEM (26,70±2,45 x 28,75±1,29), DRS (132,45 ±10,77 x 140,55±3,72), Fluência verbal: FAS (22,40±12,40 x 38,75±9,11) e Verbos (8,50±6,63 x 15,40±6,22); ED ordem direta (4,95±0,82 x 6,15±1,42), STROOP (4,40±4,87 x 0,50±0,68), BAF (12,95±2,85 x 16,25±1,25) e BCB: M2 (9,20±0,69 x 9,75±0,44), M5 (8,45±1,35 x 9,50 ±0,76). O diagnóstico de demência foi realizado em dois pacientes. Houve correlação entre o comprometimento cognitivo e a escala de RM (r=0,5348). Essa correlação foi mais forte com as alterações de hipersinal e atrofia (r=0,718). A avaliação por SPM mostrou uma hipoperfusão no caudado de forma simétrica, no cerebelo à esquerda e insula à esquerda. Conclusões: Pacientes com DW apresentam um comprometimento cognitivo, especialmente nas funções executivas, com boa correlação entre cognição e RM / Background: Wilsons disease (WD) is a rare, genetic and systemic disease, caused by a deficit in the copper metabolism, leading to its accumulation in different organs, including the brain, especially the basal ganglia. Although dystonia and dysarthria are the most common neurological signals, cognitive changes, including dementia, may emerge in untreated cases. Objective: To assess cognitive functioning of a group of WD patients and correlate with MRI and SPECT findings. Methods: All WD patients consecutively attended in a Movement Disorders Clinic between October 2006 and August 2007 were submitted to the Mini-Mental State Examination (MMSE), Brief Cognitive Screening Battery (BCSB), verbal fluency tests (animals, FAS and verbs), the Frontal Assessment Battery (FAB), STROOP, Digit Span forward and backward (DS), Dementia Ranting Scale (DRS), Wisconsin Card Sorting Test (WCST), Hooper and WAIS block design. After the tests, a MRI and SPECT were done in all patients. MRI abnormalities (high intensity signal, low intensity signal and atrophy) were ratted in a scale (0-17). SPECT was analyzed with SPM. Patients with depression or with anarthria were excluded. Results were compared with the performance of a group of healthy controls. Results: 20 WD patients (11 men) and 20 controls (9 men) were evaluated. Mean age in the WD and control groups were 30.05±7.25 years and 32.15±5.37, respectively. Mean educational level was 11.15±3.73 years among WD cases and 10.08 ± 2.62 years among controls. The WD patients had a cognitive impairment in the: MMSE (26.70±2.45 x 28.75±1.29), DRS (132.45 ±10.77 x 140.55±3.72), verbal fluency: FAS (22.40±12.40 x 38.75±9.11), Verbs (8.50±6.63 x 15.40±6.22); DS forward (4.95±0.82 x 6.15±1.42), STROOP (4.40±4.87 x 0.50±0.68), FAB (12.95±2.85 x 16.25±1.25) and BCSB: M2 (9.20±0.69 x 9.75±0.44), M5 (8.45±1.35 x 9.50 ±0.76). Dementia diagnostic was made in tow patients. There was a correlation between cognitive impairment and MRI scale (r=0.5348), this correlation was better with high intensity signal plus atrophy (r=0.718). The SPM showed a symmetrical caudate, left cerebellar and left insular hypoperfusion. Conclusion: WD patients presented cognitive impairment, especially in executive functions, with good correlation between cognition and MRI

Un problème majeur du cancer est l'apparition de métastases, difficiles à détecter par l'imagerie médicale et qui peuvent progresser rapidement. Par le biais de la modélisation mathématique, nous espérons développer de nouveaux outils capables d'anticiper l'état métastatique d'un patient.Les deux premières parties de cette thèse sont dédiées au développement d'un tel outil, l'objectif étant sonutilisation chez l'animal voire en clinique. Dû aux variabilités intra- et inter-individuelles, nous sommes amenés à utiliser des modèles statistiques coûteux en temps de calcul.Dans la partie 1, nous étendons une approche introduite par Iwata et al. et développée dans l'équipe. Nousproposons une résolution numérique plus efficace basée sur la reformulation du modèle sous formed'équation intégrale de Volterra de type convolution, qui s'avère également utile pour montrer despropriétés théoriques du modèle. En outre, nous étudions une extension stochastique de ce modèle déterministe.Dans la partie 2, nous montrons que notre approche est adaptée à la description de données souris. Utilisant le cadre statistique des modèles nonlinéaires à effets mixtes, nous construisons un modèle métastatique identifiable à partir des données et nous interprétons les résultats biologiquement.La partie 3 regroupe des résultats issus de collaborations avec des biologistes. Nous avons commencé àmodéliser la croissance tumorale à partir d'observations par imagerie SPECT en utilisant un modèle deGyllenberg et Webb. D'autre part, afin d'améliorer la précision des observations SPECT, nous testons des techniques dedétection de contours via des méthodes volumes finis basées sur des schémas DDFV. / Metastasis is one of the major problems of cancer because metastases areoften difficult to detect by clinical imaging and may develop rapidly. With the help of mathematical modelling, we hope to developnew tools capable of anticipating the metastatic state of a patient.The first two parts of this thesis are dedicated to developing such a tool, destined for a preclinical oreven clinical use. As tumour growth dynamics vary strongly between individuals and since observations are often sparse andnoisy, we need to consider computationally expensive statistical tools.In the first part, we extend an approach introduced by Iwata et al. and developed by Barbolosi et al. In particular, wepropose a more efficient numerical resolution based on a model reformulation into a Volterra integral equation of convolutiontype. This reformulation also permits to prove theoretical model properties (regularity and identifiability). Moreover, we study a stochastic generalisation of this deterministic model.In the second part, we will show that our approach is suitable for the description of experimental data on tumour-bearing mice.Using the statistical framework of nonlinear mixed-effects modelling, we build a metastatic model that is identifiable fromour data. We then interpret the results biologically.The last part of this thesis contains several results obtained in collaboration with biologists. We have started to model tumourgrowth with data obtained from SPECT imaging, using a model by Gyllenberg and Webb. Also, in order to improve the precision ofSPECT data, we have tested contour detection methods via finite volume methods based on DDFV schemes.