Made available in DSpace on 2014-10-09T12:28:33Z (GMT). No. of bitstreams: 0 / Made available in DSpace on 2014-10-09T13:56:00Z (GMT). No. of bitstreams: 0 / Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) / Tese (Doutoramento) / IPEN/T / Instituto de Pesquisas Energeticas e Nucleares - IPEN-CNEN/SP / FAPESP:08/00723-4

Made available in DSpace on 2014-10-09T12:28:54Z (GMT). No. of bitstreams: 0 / Made available in DSpace on 2014-10-09T13:57:01Z (GMT). No. of bitstreams: 0 / Dissertacao (Mestrado) / IPEN/D / Instituto de Pesquisas Energeticas e Nucleares - IPEN-CNEN/SP

Submitted by Claudinei Pracidelli (cpracide@ipen.br) on 2015-05-20T17:36:43Z No. of bitstreams: 0 / Made available in DSpace on 2015-05-20T17:36:43Z (GMT). No. of bitstreams: 0 / Tese (Doutorado em Tecnologia Nuclear) / IPEN/T / Instituto de Pesquisas Energeticas e Nucleares - IPEN-CNEN/SP

Made available in DSpace on 2014-10-09T12:28:33Z (GMT). No. of bitstreams: 0 / Made available in DSpace on 2014-10-09T13:56:00Z (GMT). No. of bitstreams: 0 / Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) / A importância da modulação da inflamação no tratamento de doenças inflamatórias bem como a dificuldade na determinação dos parâmetros de irradiação laser levou-nos a estudar os efeitos de diferentes protocolos de fototerapia com laser em baixa intensidade (potência, energia, local e momento de irradiação) no tratamento e prevenção do processo inflamatório agudo utilizando o modelo experimental de edema de pata induzido por carragenina (CGN) em três linhagens de camundongos (Balb-c, SWISS e C57BL/6). A primeira etapa do trabalho avaliou diferentes combinações de energia (1J e 3J) com diferentes potências (30, 60 e 100mW) em camundongos Balb-c irradiados na pata 1 e 2h após a injeção da CGN. A segunda etapa estudou diferentes combinações de local (pata, linfonodo inguinal e ambos) e momentos de irradiação (2 e 1h antes, 1h e imediatamente antes da CGN, 1 e 2h após e 3,5 e 4,5h após a CGN) utilizando parâmetros de irradiação fixos (1J, 100mW , 35J/cm2, área do spot 0,028cm2). A terceira etapa comparou diferentes linhagens de camundongos (Balb-c e C57Bl/6) nos melhores parâmetros de local e momento encontrados na etapa 2. Em todas as etapas foi avaliada a alteração de volume da pata através da pletismografia e o infiltrado inflamatório por histomorfometria ou análise da mieloperoxidase (MPO). Os resultados mostraram que a fototerapia com laser preveniu e tratou o edema assim como modulou o processo inflamatório com irradiações na pata e no linfonodo inguinal, de acordo com os parâmetros e a linhagem utilizada. / Tese (Doutoramento) / IPEN/T / Instituto de Pesquisas Energeticas e Nucleares - IPEN-CNEN/SP / FAPESP:08/00723-4

Made available in DSpace on 2014-10-09T12:28:54Z (GMT). No. of bitstreams: 0 / Made available in DSpace on 2014-10-09T13:57:01Z (GMT). No. of bitstreams: 0 / Neste estudo clínico verificaram-se os efeitos do laser em baixa intensidade na prevenção e tratamento da mucosite oral radio e/ou radioquimioinduzida. Foram selecionados 31 pacientes portadores de câncer em cabeça e pescoço a serem submetidos à radioterapia ou a radioterapia em associação a quimioterapia, os quais foram distribuídos aleatoriamente em três grupos: grupo 1 (controle) tratamento medicamentoso; grupo 2 tratamento medicamentoso e laserterapia diária, a partir do início da ulceração referente à mucosite grau 2 e grupo 3 tratamento medicamentoso e laserterapia diária a ser iniciada imediatamente antes do início da radioterapia. Os parâmetros de irradiação foram: comprimento de onda de 660nm, potência de 100mW, modo de operação contínuo, aplicação pontual, energia de 2J por ponto em 30 pontos pré-determinados, sendo o tempo utilizado de 20s por ponto. O grupo controle recebeu tratamento medicamentoso que consistia no uso de um conjunto de medidas preventivas e terapêuticas para abordagem dos efeitos adversos agudos radioinduzidos. Os resultados foram avaliados, quanto à ocorrência, graus e escore de dor associados à mucosite oral, perda de massa corpórea, uso de sonda nasogástrica, necessidade de internação e interrupção do tratamento oncológico decorrentes da mucosite oral. Os resultados indicam que o protocolo de aplicação do laser em sua forma preventiva é o mais efetivo na prevenção e tratamento da mucosite oral e que seu uso diário contribuiu para o alívio da sintomatologia dolorosa colaborando para melhora da qualidade de vida do paciente oncológico. / Dissertacao (Mestrado) / IPEN/D / Instituto de Pesquisas Energeticas e Nucleares - IPEN-CNEN/SP

Submitted by Claudinei Pracidelli (cpracide@ipen.br) on 2015-05-20T17:36:43Z No. of bitstreams: 0 / Made available in DSpace on 2015-05-20T17:36:43Z (GMT). No. of bitstreams: 0 / A radioterapia é uma das três principais modalidades utilizadas no tratamento de doenças malignas como o câncer; as outras duas são a quimioterapia e a radiocirurgia. Em contraste com outras especialidades médicas que necessitam principalmente do conhecimento clínico e da experiência de especialistas, a radioterapia, com a utilização da radiação ionizante no tratamento do câncer, depende do investimento pesado em tecnologias modernas e dos esforços colaborativos de diversos profissionais, cuja equipe coordenada influencia, sobremaneira, o resultado do tratamento. A Radioterapia de intensidade modulada (IMRT) com o uso de colimadores multilâminas (multileaf collimators MLCs) tem o potencial para alcançar um alto grau de conformidade da dose no alvo (tumor a ser tratado) e ainda promover a proteção de tecidos normais do que a maioria de outras técnicas de tratamento, especialmente para volumes-alvo ou órgãos de risco com formatos complexos. Entretanto, estudos recentes mostraram que baixas doses de radiação podem causar tumores secundários. Esse trabalho tem como objetivo a determinação da distribuição de dose de radiação absorvida em diversas simulações de tratamentos radioterápicos com o uso de dosímetros compostos de LiF:Mg,Ti; CaSO4:Dy e Al2O3:C, utilizando um objeto simulador de polimetilmetacrilato (PMMA) empregando as seguintes técnicas dosimétricas: termoluminescência (TL), luminescência opticamente estimulada (OSL) e detectores semicondutores (diodos). / Tese (Doutorado em Tecnologia Nuclear) / IPEN/T / Instituto de Pesquisas Energeticas e Nucleares - IPEN-CNEN/SP

La vie sur Terre s’est adaptée à un environnement où il y a un faible et persistent bruit de fond radiatif qui interagit avec les cellules. Loin des effets clairement nocifs des radiations à haute dose, il est difficile d’évaluer et de comprendre les impacts des faibles doses de la radioactivité naturelle sur les systèmes vivants. Nous avons tenté d’étudier si le bruit de fond radiatif est un facteur important dans l’évolution, en menant des expériences évolutives identiques avec Escherichia coli au Laboratoire de Physique Corpusculaire de Clermont-Ferrand, et au Laboratoire Souterrain de Modane. Malgré une différence d’un facteur 7,3 entre les taux d’interaction des rayonnements ionisants avec les cellules dans les deux laboratoires, aucune différence significative n’a pu être trouvée dans le fitness compétitif des populations cellulaires évoluées dans chaque laboratoire. Par simulation, nous avons montré que le taux d’interaction entre le bruit de fond radiatif et E. coli est cent fois plus faible que le taux de mutations d’origine endémique, ce qui renforce l’hypothèse que les radiations naturelles ont peu d’effet sur l’évolution. Dans le cadre du projet Geant4-DNA, nous avons développé une application complète de simulation mécanistique des dommages radio-induits à l’ADN, afin d’explorer davantage cette hypothèse. Avec cette application, on a irradié un modèle du génome d’E. coli, montrant que pour l’irradiation par des électrons d’énergies > 10 keV, le rendement des cassures double brin est de 0,006 – 0,010 CDB Gy-1 Mbp-1, selon le modèle de piégeage des radicaux chimiques. Ce résultat est en accord avec des données expérimentales, et souligne plus encore que les radiations ionisantes d’origine naturelle n’ont qu’une contribution mineure aux mutations responsables de l’évolution. / All life on earth has adapted to an environment where there is a small, persistent, radiation background interacting with cells. Unlike evaluating the clearly harmful effects of high radiation doses, understanding the effects of this low persistent radiation dose on living systems is incredibly difficult. We have attempted to study whether background radiation is an important factor in evolution by conducting identical evolution experiments with Escherichia coli in the Clermont-Ferrand Particle Physics Laboratory and the Modane Underground Laboratory. Despite a 7.3 fold difference in the rate of interactions between the radiation background and cells between the two environments, no significant difference was found in the competitive fitness of the cell populations grown at each location. Using simulations, we showed that the rate at which ionising radiation interacts with cells is one hundred times less frequent than E. coli’s mutation rate in our experimental conditions, supporting the contention that natural radiation has no strong evolutionary effect. To further support this conclusion, we developed a mechanistic simulation for DNA damage as part of the Geant4-DNA project. Using this application, we irradiated a model of an E. coli genome, showing that for electron irradiation > 10 keV, the double strand break yield can be reasonably estimated to be between 0.006 – 0.010 DSB Gy-1 Mbp-1, depending upon the modelling of radical scavenging. This is in agreement with experimental data, further highlighting the small role natural ionising radation plays as a cause of mutations.

Patients in the intensive and critical care unit frequently undergo diagnostic radiology procedures such as computed tomography (CT) and X-ray imaging. As these patients often require respiratory assistance and are vulnerable to infection, it is important to understand the potential acute effects of these procedures on the lungs and immune system. The aim of this study was to determine the acute effects of a single clinically relevant low-dose X-ray exposure in order to establish baseline responses in markers of lung injury and immune function in a rodent model. Male Sprague-Dawley rats (200-250 g) were irradiated with 0, 2, 20 or 200 mGy whole-body X-rays in an XRAD 320 irradiator. Markers of lung injury and immune activation in the lungs and spleen were evaluated 0.5, 4, and 24 h post-irradiation to examine the acute stages of the physiological and immunological response. Intratrachaeal lipopolysaccharide (LPS) exposure was used as a positive control model of acute lung injury. Lung injury endpoints included respiratory mechanics, pulmonary oedema, arterial blood oxygenation, histological analysis, and cellular and proteinaceous infiltrate via bronchoalveolar lavage. Immunological measures in the spleen focused on splenocyte proliferation, using the MTS assay and differential cell counts before and after stimulation with LPS or concanavalin A (Con A), as compared to unstimulated cultures. Splenocyte proliferation in response to Con A, but not LPS, was significantly decreased after 200 mGy in vivo X-irradiation (repeated measures two-way ANOVA with LSD post-hoc, p=0.024). There was a non-significant trend towards increased lung tissue resistance after 200 mGy, with no significant effect on pulmonary oedema, cellular or proteinaceous infiltrate, nor other aspects of respiratory mechanics (two-way ANOVA with LSD post-hoc, p>0.05). A clear understanding of these immunological and physiological effects informs the responsible use of medical diagnostic procedures in modern medicine. Establishment of this model for the elucidation of acute immune effects of low-dose radiation will facilitate future work evaluating these parameters in disease models, mimicking patients in intensive care. / Thesis / Master of Science (MSc) / Diagnostic procedures such as computed tomography (CT) and X-ray imaging are a common part of intensive and critical care medicine. Some physicians are concerned that this exposure to diagnostic radiation may negatively affect the health of their patients, who are prone to infection and who often need a machine to breathe for them. In order for doctors to make informed decisions, the possible effects of these levels of radiation must be understood. To improve this understanding, this study looked at the short-term effects of X-ray doses on key organs affected by critical illness, the lungs, and the spleen, which is an important organ of the immune system that helps fight infection. Using an animal model, doses of X-rays in the range of diagnostic radiation (0-200 mGy) were examined and no significant effect on lung health was found. However, the highest dose of X-rays tested, which is greater than that expected for a single CT scan, did have an effect on cells from the spleen. Spleen cells are designed to multiply when they detect various types of infection, so that there are more immune cells to fight that infection. The cells from animals that were given the highest dose of X-rays didn’t multiply as much in response to infective stimulus as those from animals that received lower doses, or no X-rays at all. Overall, it seems that diagnostic radiation doesn’t have an effect in the lungs, but very high diagnostic doses could slightly affect a patient’s ability to fight infection. It is important to remember that patients in critical care are very sick, so doctors have good reason to use diagnostic tools available to them. Missing a diagnosis has major and immediate consequences, which must be balanced against the potential small risks of using radiation to make that diagnosis.

Anti-inflammatory low-dose therapy is well established, whereas the immunomodulatory impact of doses below 0.1 Gy is much less clear. In this study, we investigated dose, dose rate and time-dependent effects in a dose range of 0.005 to 2 Gy on immune parameters after whole body irradiation (IR) using a pro-inflammatory (ApoE−/−) and a wild type mouse model. Long-term effects on spleen function (proliferation, monocyte expression) were analyzed 3 months, and short-term effects on immune plasma parameters (IL6, IL10, IL12p70, KC, MCP1, INFγ, TGFβ, fibrinogen, sICAM, sVCAM, sE-selectin/CD62) were analyzed 1, 7 and 28 days after Co60 γ-irradiation (IR) at low dose rate (LDR, 0.001 Gy/day) and at high dose rate (HDR). In vitro measurements of murine monocyte (WEHI-274.1) adhesion and cytokine release (KC, MCP1, IL6, TGFβ) after low-dose IR (150 kV X-ray unit) of murine endothelial cell (EC) lines (H5V, mlEND1, bEND3) supplement the data. RT-PCR revealed significant reduction of Ki67 and CD68 expression in the spleen of ApoE−/− mice after 0.025 to 2 Gy exposure at HDR, but only after 2 Gy at LDR. Plasma levels in wild type mice, showed non-linear time-dependent induction of proinflammatory cytokines and reduction of TGFβ at doses as low as 0.005 Gy at both dose rates, whereas sICAM and fibrinogen levels changed in a dose rate-specific manner. In ApoE−/− mice, levels of sICAM increased and fibrinogen decreased at both dose rates, whereas TGFβ increased mainly at HDR. Non-irradiated plasma samples revealed significant age-related enhancement of cytokines and adhesion molecules except for sICAM. In vitro data indicate that endothelial cells may contribute to systemic IR effects and confirm changes of adhesion properties suggested by altered sICAM plasma levels. The differential immunomodulatory effects shown here provide insights in inflammatory changes occurring at doses far below standard anti-inflammatory therapy and are of particular importance after diagnostic and chronic environmental exposures.

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