These studies investigated the impact of intensity and body temperature on performance and cardiovascular regulation during high intensity and prolonged exercise. In study 1, polyphenol antioxidant supplementation proved to have no effect on exercise performance and related variables (gross efficiency, perceived exertion, maximal power) during exercise in the heat. Furthermore, there were no differences between the cardiovascular or thermoregulatory responses between control and antioxidant treatments. Study 2 utilized an integrative approach to investigate a classic topic in exercise physiology, namely, is the cardiac output to oxygen consumption relationship linear across a wide range of exercise intensities? The slope of the CO vs. VO2 relationship was significantly reduced from 70 to 100% of VO2max when compared to the slope from 40 to 70% of VO2max (2.0 ± 0.4 vs. 4.4 ± 0.3 l/min, p = 0.025). This finding, in combination with the plateau and eventual reduction in stroke volume at high intensity exercise compared to moderate intensity exercise (146.0 ± 16.6 vs. 138.5 ± 14.9 ml/beat, p = 0.015), argues in favor of a cardiac limitation to high intensity exercise. This study also showed that the pattern of oxygen extraction at the whole body level (arterial venous O2 difference) and the muscle level (deoxygenated hemoglobin) is not similar and that muscle specific differences exist regarding oxygen extraction. Study 3 determined that hyperthermia (elevation of skin temperature by 4.3°C and core temperature by 0.8°C) did not reduce SV independent of the increase in HR. Even under conditions of moderate hyperthermia the reduction in SV is due to the increase in HR and temporally unrelated to increases in cutaneous blood flow. In summary, antioxidant supplementation had no effect on performance, cardiovascular, or thermoregulatory responses to exercise in the heat in well trained subjects. High intensity exercise is associated with a reduced rate of increase in the CO vs. VO2 relationship. Finally, hyperthermia does not reduce SV during exercise when HR is maintained at normal levels. / text

The primary purpose of this work was to evaluate the capability of nanoparticles to transform electromagnetic energy at microwave frequencies into therapeutic heating. Targeted nanoparticles, in conjunction with microwave irradiation, can increase the temperatures of the targeted area over the peripheral region. Therefore, to become clinically viable, microwave absorbing nanoparticles must first be identified, and a system to monitor the treatment must be developed. In this study, ultrasound temperature imaging was used to monitor the temperature of deep lying structures. First, a material-dependent quantity to correlate the temperature induced changes in ultrasound images (i.e. apparent time shifts) to differential temperatures was gathered for a tissue-mimicking phantom, porcine longissimus dorsi muscle, and porcine fat. Then microwave nanoabsorbers were identified using an infrared radiometer. The determined nanoabsorbers were then injected into ex-vivo porcine longissimus dorsi muscle tissue. Ultrasound imaging frames were gathered during microwave treatment of the inoculated tissue. Finally, the ultrasound frames were analyzed using the correlation between temperature and apparent shifts in ultrasound for porcine muscle tissue. The outcome was depth-resolved temperature profiles of the ex-vivo porcine muscle during treatment. The results of this study show that magnetite is a microwave nanoabsorber that increases the targeted temperature of microwave hyperthermia treatments. Overall, there is clinical potential to use microwave nanoabsorbers to increase the efficiency of microwave hyperthermia treatments. / text

Iron oxide nanoparticles are highly researched for their use in biomedical applications such as drug delivery, diagnosis, and therapy. The inherent biodegradable and biocompatible nanoparticle properties make them highly advantageous in nanomedicine. The magnetic properties of iron oxide nanoparticles make them promising candidates for magnetic fluid hyperthermia applications. Designing an efficient iron oxide nanoparticle for hyperthermia requires synthetic, surface functionalization, stability, and biological investigations. This research focused on the following three areas: optimizing synthesis conditions for maximum radiofrequency induced magnetic hyperthermia, designing a simple and modifiable surface functionalization method for specific or broad biological stability, and in vitro and in vivo testing of surface functionalized iron oxide nanoparticles in delivering effective hyperthermia or radiotherapy. The benzyl alcohol modified seed growth method of synthesizing iron oxide nanoparticles using iron acetylacetonate as an iron precursor was investigated to identify significant nanoparticle properties that effect radiofrequency induced magnetic hyperthermia. Investigation of this synthesis under atmospheric conditions revealed a combination of thermal decomposition and oxidation-reduction mechanisms that can produce nanoparticles with larger crystallite sizes and decreased size distributions. Nanoparticles were easily surface functionalized with (3-Glycidyloxypropyl)trimethoxysilane (GLYMO) without the need for organic-aqueous phase transfer methods. The epoxy ring on GLYMO facilitated post-modifications via a base catalyzed epoxy ring opening to obtain nanoparticles with different terminal groups. Glycine, serine, γ-aminobutryic acid (ABA), (S)-(-)-4-amino-2-hydroxybutyric acid (SAHBA), ethylenediamine, and tetraethylenepentamine were successful in modifying GLYMO coated-iron oxide nanoparticles to provide colloidal and varying biological stability while also allowing for further conjugation of chemotherapeutics or radiotherapeutics. The colloidal stability of cationic and anionic nanoparticles in several biologically relevant media was studied to address claims of increased cellular uptake for cationic nanoparticles. The surface functionalized iron oxide nanoparticles were investigated to determine effects on cellular uptake and viability. In vitro tests were used to confirm the ability of iron oxide nanoparticles to provide effective hyperthermia treatment. S-2-(4-Aminobenzyl)-1,4,7,10-tetraazacyclododecane tetraacetic acid (DOTA) was coupled to SAHBA and carboxymethylated polyvinyl alcohol surface functionalized iron oxide nanoparticles and radiolabeled with 177Lu. The capability of radiolabeled iron oxide nanoparticles for delivering radiation therapy to a U87MG murine orthotopic xenograft model of glioblastoma was initially investigated.

Doctor of Philosophy / Department of Chemistry / Stefan H. Bossmann / Cancer is one of the leading causes of death in the world. Billions of dollars are spent to treat cancer every year. This clearly shows the need for developing improved treatment techniques that are affordable to every person. Early diagnosis and imaging of tumors is equally important for the battle against this disease. This dissertation will discuss new approaches for discovering and developing novel detection and treatment techniques for cancer using organic ligands, and Fe/Fe3O4 core/shell magnetic nanoparticles. A series of o-phenylenediamine derivatives with nitro-, methyl- and chloro- substituents were synthesized and studied their ability to act as anticancer agents by using steady-state, UV/Vis-, and fluorescence spectroscopy. In the absence of zinc(II), intercalation with DNA is the most probable mode of interaction. Upon addition of zinc(II), DNA-surface binding of the supramolecular aggregates was observed. The interaction of the supramolecular (-ligand-Zn2+-)n aggregates with MDA 231 breast cancer cells led to significant cell death in the presence of UVA at λ=313 nm displaying their potential as anticancer agents. Bimagnetic Fe/Fe3O4 core/shell nanoparticles (MNPs) were designed for cancer targeting after intratumoral or intravenous administration. Their inorganic center was protected by dopamine-oligoethylene glycol ligands. TCPP (4-tetracarboxyphenyl porphyrin), a fluorescent dye, was attached to the dopamine-oligoethylene glycol ligands. These modified nanoparticles have the ability to selectively accumulate within the cancerous cells. They are suitable candidates for local hyperthermia treatment. We have observed a temperature increase of 11 ºC in live mice when subcutaneously injecting the MNPs at the cancer site and applying an alternating magnetic field The system is also suitable for Magnetic Resonance Imaging (MRI), which is a diagnostic tool to obtain images of the tumors. Our superparamagnetic iron oxide nanoparticles have the ability to function as T1 weighted imaging agents or positive contrasting agents. We were able to image tumors in mice using MRI. Various proteases are over-expressed by numerous cancer cell lines and, therefore, of diagnostic value. Our diagnostic nanoplatforms, designed for the measurement of protease activities in various body fluids (blood, saliva, and urine), comprise Fe/Fe3O4 core/shell nanoparticles featuring consensus sequences, which are specific for the target protease. Linked to the consensus sequence is a fluorescent organic dye (e.g. TCPP). Cleavage of the sequence by the target protease can be detected as a significant increase in fluorescence occurring from TCPP. We were able to correlate our diagnostic results with cancer prognosis.

Doctor of Philosophy / Department of Chemistry / Stefan H. Bossmann / Recent development of a variety of superparamagnetic and ferromagnetic iron/iron oxide (Fe/Fe₃O₄) nanoparticles with different surface chemistry have been widely studied for numerous biological applications such as drug delivery, as diagnostics, hyperthermia and magnetic resonance imaging. The wide applications of Fe/Fe₃O₄ nanoparticles are possible since they exhibit favorable properties as high magnetization ability, are smaller than 100 nm in size, they can be coated with several ligands which allow drug delivery at a specific site and are biocompatible. By using Fe/Fe₃O₄ nanoparticles as drug delivery agents treatment costs and side effects can be reduced, however treatment efficacy can be increased. We have demonstrated that Fe/Fe₃O₄ nanoparticles can be utilized in different methods depending on their properties, to be used as therapeutic agents for cancer treatment. In one method we have taken advantage of the Fe/Fe₃O₄ nanoparticles magnetic ability to produce hyperthermia (heat) in cancer cells when subjected to an alternative magnetic field. Here we use the cell based delivery system since the size of the nanoparticles are small they can be taken up by monocyte/ macrophage like cells for systemic transportation to the inflamed cancer cite. The hyperthermia study was conducted in mice with pancreatic cancer. This study demonstrated that the life expectancy of the mice increased by 31%. In the next method we took the advantage of the surface chemistry of the Fe/Fe₃O₄ nanoparticles and changed it with dopamine-peptide and dopamine-thiosemicarbazone ligands. The advantage of the peptide is to deliver the nanoparticle to its target site and the thiosemicarbazone analogue is used as an iron chelator that would initiate apoptosis in cancer cells. This nanoplatform was tested in 4T1 breast cancer cell line and normal fibroblast cell line and demonstrated that it was effective towards the cancer cell line than the normal cell line at a ratio of 5:1 of thiosemicarbazone analogue : dopamine on the nanoparticle. However further studies are needed to be done to clarify the effectiveness of this nanosystem.

Sabe-se que o aumento na temperatura do tecido tumoral pode aumentar a eficiência de técnicas convencionais de combate ao câncer (radioterapia e quimioterapia). Além disso, a variação de temperatura em tumores pode ser uma forma de tratamento alternativo à cirurgia, feito por meio do fornecimento de calor direcionado às células cancerosas e preservando o tecido sadio. Para maior eficácia e segurança no emprego de técnicas que utilizam fornecimento de calor ao tecido biológico, é necessário o monitoramento da temperatura tecidual para garantir que a morte celular por ablação térmica seja limitada ao tecido alvo, minimizando os danos aos tecidos adjacentes. A imagem fotoacústica é uma técnica baseada no efeito fotoacústico, o qual consiste na absorção de radiação eletromagnética pelo tecido e, devido à expansão termoelástica, na geração de ondas acústicas. A amplitude da onda de pressão gerada pelo efeito fotoacústico possui dependência com a temperatura do meio pelo parâmetro de Grueneisen, que depende das propriedades mecânicas do material. Portanto, mudanças na amplitude do sinal fotoacústico carregam informações a respeito da variação na temperatura do material. Neste trabalho, a dependência da amplitude do sinal fotoacústico com a temperatura foi estudada em um material simulador de tecido biológico (phantom) em condições similares a de tratamentos por hipertermia Nesse caso, imagens fotoacústicas foram adquiridas para cada grau de temperatura em uma faixa de 36 até 41 ºC durante o procedimento de aquecimento por banho térmico. Mudanças na amplitude e fase do sinal fotoacústico foram avaliadas através da aplicação de algoritmos de speckle tracking. Para estimar a variação na amplitude do sinal também foram utilizados e avaliados diferentes métodos de comparação. Os resultados são apresentados por imagens fotoacústicas termais produzidas pela aplicação de um fator de calibração aos mapas de variação relativa da amplitude do sinal em função da temperatura do meio. Finalmente, avaliamos um experimento de hipertemia por ultrassom focalizado de alta intensidade (High Intensity Therapeutic Ultrasound - HITU) em uma amostra de músculo suíno. Nesse caso foram geradas imagens termais fotoacústicas e imagens termais produzidas pela mudança de fase do sinal pulso-eco de ultrassom. Os resultados sugerem uma maneira não invasiva de calcular a distribuição da variação de temperatura do meio que pode ser aplicada para monitoramento durante tratamentos que utilizam o fornecimento de calor ao tecido biológico. / Several studies have shown that elevating the temperature of tumoral tissue improves standard cancer treatments success rate (radiotherapy and chemotherapy). This procedure can also be a therapy to cancer by delivering heat and killing cancer cells while healthy tissues are preserved. For improved efficiency and security in heat applications, it is important to monitor tissue temperature during treatments. Photoacoustic (PA) pressure wave amplitude has a temperature dependence given by the sample mechanical properties (Gruenesein parameter). These changes in photoacoustic signal amplitude carry information about temperature variation in tissue. Therefore, PA has been proposed as an imaging technique to monitor temperature during hyperthermia. In this study, PA images were acquired for temperatures ranging from 36ºC to 41ºC using a tissue-mimicking phantom immersed in a temperature controlled thermal bath. Relative amplitude variation was calculated using speckle tracking algorithms using four different methods to estimate these variations in PA signal amplitude. The results are presented as PA-based thermal images, generated using a calibration factor to the percentage variations in the amplitude maps. Finally, PA-based and ultrasound-based thermal images were acquired during heating by high intensity focused ultrasound (High Intensity Therapeutic Ultrasound - HITU) in a porcine muscle. The results suggest a non-invasive way to monitor temperature during hyperthermia procedures.

Nanopartículas de ouro têm mostrado enorme potencial de aplicação em modalidades diagnósticas e terapêuticas fotoativadas. Em especial, nanoestruturas de ouro anisotrópicas ramificadas apresentam excelente desempenho atuando tanto como contrastes de imagens fotoacústicas, quanto como agentes ativos para terapias fototérmicas de câncer. Apesar dos avanços nas suas rotas de síntese, o desenvolvimento dessas nanoestruturas de forma simples e reprodutível ainda é desafiador. O presente trabalho visou o desenvolvimento de nanopartículas de ouro anisotrópicas ramificadas, ou nanoflores, que sejam fotoativas no infravermelho-próximo para a terapia e diagnóstico de câncer. Em particular, buscou-se o desenvolvimento de uma síntese simples para sua obtenção, assim como a verificação de sua atuação como agente de contraste fotoacústico e como agente ativo para hipertermia de tumores. Para tanto, desenvolveu-se uma síntese in situ que permitiu a obtenção de nanoflores monodispersas com tamanho e propriedades ópticas controláveis. Através da variação de aspectos da síntese, como a temperatura e a concentração de ouro, foi possível sintonizar a atividade óptica das partículas entre 590 e 960 nm. Sua formação foi confirmada por microscopia eletrônica de varredura, espalhamento de luz dinâmico e espectroscopia UV-visível. As partículas apresentaram boa estabilidade de suas características físico-químicas por dois meses e meio. Ainda, as nanoflores se mostraram estáveis, também, quando suspensas em meio de cultura, sob irradiação de lasers, e quando mantidas a temperatura corpórea por longos intervalos. Sua resposta fotoacústica foi caracterizada, apresentando sinais significativos e permitindo a obtenção de imagens claras de sua localização, mesmo em baixas concentrações. Testes realizados em cultura de células mostraram que as nanoflores foram eficazes na hipertermia de uma linhagem de hepatocarcinoma de rato (HTC), ao mesmo tempo que não apresentaram sinais de toxicidade a uma linhagem de fibroblastos de camundongos (FC3H). Esses resultados revelam uma possibilidade simples de fabricação de nanoestruturas de ouro anisotrópicas ramificadas, que podem servir como uma plataforma promissora para o diagnóstico e terapia do câncer. / Gold nanoparticles have shown enormous potential of application in photodiagnostic and in phototherapeutic procedures. Notably, branched anisotropic gold nanostructures present distinguished performance acting as contrast agents of photoacoustic images and as active agents for photothermal therapies for cancer. Despite advances in their synthesis routes, the growth of these nanostructures in a simple and reproducible way is still challenging. The present study was aimed at developing branched anisotropic gold nanoparticles, coined nanoflowers, that are photoactive in the near-infrared for therapy and diagnosis of cancer. In particular, we sought to develop a simple synthesis route, as well as to verify its application for both, as photoacoustic contrast agents and as active agents for tumor hyperthermia. An in situ synthesis was developed which allowed the development of monodisperse nanoflowers with controllable size and optical properties. Through variations of certain aspects of this procedure, such as temperature and gold ions concentration, it was possible to tune the optical activity of the particles between 590 and 960 nm. The nanostructure morphology was confirmed by scanning electron microscopy, dynamic light scattering and UV-visible spectroscopy. The particles exhibited consistent physicochemical characteristics and good stability for two and a half months. Furthermore, the nanoflowers were also stable when suspended in cell culture medium, under laser irradiation and when maintained at body temperature for long intervals. Its photoacoustic response was characterized, presenting significant responses and generating clear images of its location, even at low concentrations. In vitro tests revealed that these nanoflowers were effective therapeutic agents for photothermal therapy of a rat hepatocarcinoma (HTC) lineage, while showing no signs of toxicity to mouse fibroblast (FC3H) cell line. These results reveal a simple procedure of synthesizing branched anisotropic gold nanostructures, which can serve as a promising platform for cancer diagnosis and therapy.

Estudos com tratamento hipertérmico de tumores utilizando nanopartículas metálicas têm sido realizados durante as últimas décadas e mostram resultados bons quanto à remissão de tumores, por vezes chegando à cura completa. O mesmo acontece em relação aos tratamentos baseados em ação fotodinâmica de fotossensibilizadores. Tratamentos aliando a terapia hipertérmica com nanopartículas de ouro e a terapia fotodinâmica com diversos fotossensibilizadores tem efeito sinérgico e apresenta excelente potencial terapêutico, em que pese serem necessários mais estudos para que uma nova terapia conjunta possa ser implementada. A proposta deste trabalho foi investigar esse efeito sinérgico utilizando nanobastões de ouro complexados com fotossensibilizadores. Após a síntese dos nanobastões pelo método de seeding, a eficácia do tratamento fotodinâmico e da terapia hipertérmica, separadamente, foi investigada. A metodologia do recobrimento dos nanobastões por fotossensibilizador, em um primeiro momento, não logrou êxito com a porfirina, porém com a ftalocianina tetracarboxilada se mostrou mais eficaz. A taxa de fotodegradação da ftalocianina em solução foi investigada como parâmetro para a eficiência em geração de oxigênio singlete. Após centrifugação e lavagem das nanopartículas, no entanto, evidenciou-se por espectrofotometria que o fotossensibilizador não permaneceu aderido aos nanobastões. Em um segundo momento, optamos por recobrir os nanobastões por porfirinas tetrassulfonadas, com ou sem grupamentos metil-glucamina. Após o processo de recobrimento, essas ftalocianinas formaram complexos iônicos com o CTAB que recobre os nanobastões. Os complexos nanobastões-ftalocianinas foram analisados por microscopia eletrônica de transmissão e as taxas de geração de oxigênio singlete e de radical hidroxil foram investigadas. Além disso, foram utilizadas para testes in vivo e in vitro com células de melanoma melanótico (B16F10) ou amelanótico (B16G4F). As células tumorais em cultura ou os tumores em camundongos C57BL6 foram irradiados com luz em 635 nm e os tumores foram observados por 15 dias após o tratamento. Houve evidente aumento na geração de oxigênio singlete por ambos fotossensibilizadores, e maior geração de radicais livres por parte do fotossensibilizador metilglucaminado. O oposto ocorre com o fotossensibilizador sem metilglucamina. Houve, também, moderada citotoxicidade no escuro quando células foram incubadas com nanopartículas recobertas por ftalocianinas ou não. Quando ativados pela luz, os complexos ftalocianinas-nanobastões desencadearam um aumento de 5ºC no meio de cultura das células, e a morte celular observada foi extensa (91% para a linhagem B16G4F e 95% para a linhagem B16F10). Tanto os resultados in vitro quanto os in vivo indicam que as propriedades das ftalocianinas testadas são melhoradas significativamente quando elas estão complexadas aos nanobastões. Este é um estudo pioneiro por utilizar duas porfirinas tetrassulfonadas específicas e por utilizar o mesmo comprimento de onda para a ativação dos fotossensibilizadores e nanobastões. / Studies with hyperthermic tumor ablation using metallic nanoparticles have been performed on the last decades, and show promising results on tumor remission, sometimes achieving the complete cancer elimination. The same occurs regarding on treatments based on photodynamic activity of photosensitizer compounds. Studies indicate that those therapeutic interventions - hyperthermic therapy using gold nanorods and photodynamic activity with many photosensitizers - together can present a synergistic effect, and offer a great therapeutic potential, although more investigation needs to be performed before such approach could be implemented. We proposed to investigate the effect of the attachment of photosensitizers onto the surface of gold nanorods (well-characterized devices for hyperthermia generation). After nanorods synthesis through a seed-mediated method, the PDT and hyperthermia\'s efficacy was assessed separately. The method used for covering the gold nanorods with photosensitizers did not permit, in a first approach, the attachment of porphyrins onto the nanoparticles surface, but the attachment of tetrasulfonated phthalocyanines was more successful. The phthalocyanine\'s degradation rate was assessed as an indirect parameter of singlet oxygen generation. After centrifuging and washing the nanoparticles, we saw that the photosensitizers do not keep attached to the nanorods. On a second approach, we chose to recover the nanorods with two zinc phthalocyanines - with or without methyl-glucamine groups. After the recovering process, the phthalocyanines formed ionic complexes with the CTAB that is previously recovering the nanoparticles. The nanorod-phthalocyanines complexes were analyzed by TEM, and their singlet oxygen and hydroxyl radical generation yield were assessed. Furthermore, they were tested in vitro in melanoticB16F10 and amelanotic B16G4F melanoma cells, and in vivo. The tumor cells (in vitro) and the tumor tissue (in vivo) with nanoparticles were irradiated with laser (at 635 nm), and the tumor growth in mice was observed for 15 days after the laser irradiation. It is evident the increase in the singlet oxygen generation, and higher HPF activation for the glucaminated Pc, but the inverse for the other photosensitizer. It seems like there is a type I to type II switch on the action mechanism of the latter Pc. A mild cytoxocity was observed with the nanorods conjugated with photosensitizer in the dark, but when they are activated by light (and taken into account a 5ºC rise in the temperature because of the surface plasmon resonance from the gold nanorods), the cell killing is intense (91% for B16G4F cell line, and 95% for B16F10 cell line). Both in vitro and in vivo results indicate that the photodynamic properties of the phthalocyanines tested are enhanced when they are attached onto the nanorods surface. This is a novel study because we used two tetrasulfonated phthalocyanines and because we used the same wavelength to activate both the nanorods and the photosensitizers.

A ação combinada da Terapia Fotodinâmica (TFD) e da Hipertermia (HPT), projetadas para trabalhar sinergicamente, pode levar a uma considerável regressão de tumores neoplásicos após mínimas doses de dissipação de calor e/ou fotossensitização luminosa. Nesse estudo foi proposto, inicialmente, a formação do complexo de inclusão entre o fármaco fotossensibilizante, ftalocianina de zinco (ZnPc) e a cucurbiturila (CB[7]), de modo a aumentar a solubilidade do fármaco e minimizar sua agregação. Além disso, foi proposto nanoencapsular o complexo em lipossomas, tornando-os bicompatíveis. O complexo de inclusão CB[7]:ZnPc foi preparado pelo método de co-evaporação complementado por liofilização e caracterizado por Ressonância Magnética Nuclear (RMN 1H), Calorimetria Diferencial Exploratória (DSC) e Difração de Raios-X (XDR). As formulações lipossomais foram preparadas pelo método de hidratação do filme lipídico e caracterizadas com relação ao tamanho das vesículas, potencial Zeta, índice de polidispersão e estabilidade. Nos estudos biológicos, utilizando a linhagem neoplásica B16-F10, foi feita uma comparação entre o complexo livre e encapsulado em lipossoma, a fim de estudar o lipossoma como um veículo de liberação de fármacos. Além desse estudo, foi avaliada a toxicidade do lipossoma contendo o complexo de inclusão e o fluido magnético (CB:ZnPc-MLp) na presença de luz e campo magnético. Esse resultado nos mostra que a aplicação da luz e do campo magnético em conjunto, consegue ser mais eficaz do que os mesmos aplicados separadamente, comprovando o efeito sinérgico. / The combined action of photodynamic therapy (PDT) and hyperthermia (HPT), designed to work synergistically, may lead to a significant regression of neoplasic tumors after minimum doses of heat dissipation and / or light photosensitization. In this study was provided initially, the formation of an inclusion complex between the drug photosensitizer, zinc phthalocyanine (ZnPc) and cucurbit[7]uril (CB [7]) in order to increase the solubility of the drug and minimize aggregation. Furthermore, it was also proposed nanoencapsular liposome complex, making them biocompatible. The inclusion complex CB [7]: ZnPc was prepared by co-evaporation method complemented by lyophilization and characterized by nuclear magnetic resonance (1H-NMR), Differential Scanning Calorimetry (DSC) and X-ray diffraction (XDR). Liposomal formulations were prepared by lipid film hydration and characterized with respect to vesicle size, zeta potential, polydispersity and stability. In the biological studies, using neoplasic cell line B16-F10, a comparison was made between the free complex and the complex encapsulated in liposome in order to study the liposome as a carrier for drug delivery. In this study, we evaluated the toxicity of the liposome containing the inclusion complex and the magnetic fluid (CB: ZnPc-MLP) in the presence of magnetic field and light. This result shows that the application of light and magnetic field in combination are able to be more effective than the same applied separately, demonstrating the synergistic effect.

As células dendríticas (DCs) são células apresentadoras de antígeno suscetíveis a muitos sinais de ativação, os quais induzem diferentes padrões de ativação e resposta de linfócitos T. Neste trabalho, estudamos os efeitos de dois sinais de perigo, a febre e o LPS, sobre o fenótipo e função de DCs. A exposição de DCs ao calor não afetou a expressão de CD80 e CD86, capacidade endocítica ou produção de citocinas, características que foram afetadas pelo LPS. Entretanto, DCs expostas ao calor apresentaram uma maior atividade aloestimuladora e maior expressão de CD40. Quando DCs ativadas com LPS foram também estimuladas pelo calor, nenhuma alteração fenotípica na superfície celular foi notada, mas as DCs induziram maior produção de IFN-<font face=\"symbol\">g por linfócitos T alogenêicos e favoreceram a proliferação de linfócitos T CD8+. Esses dados indicam que a febre pode favorecer uma resposta celular, através de sua ação sobre DCs ativadas, com provável participação do CD40. Além de seu significado fisiológico, esse fenômeno pode ter aplicação em estratégias imunoterapêuticas. / Dendritic cells (DCs) are the main antigen presenting cells and susceptible to many activation signals that will induce different patterns of DC activation and of T cell responses. In this work we studied the effects of two danger signals, fever and LPS, on DC phenotype and function. Exposure of immature monocyte-derived dendritic cells to heat did not affect CD80 and CD86 expression, their endocytic ability, or their cytokine production, characteristics that were affected by LPS. However, heat-exposed DCs presented a higher allo-stimulatory activity and enhanced CD40 expression. When LPS activated DCs were also stimulated by heat, no cell surface phenotypic change was noted but these cells induced a higher IFN-<font face=\"symbol\">g secretion by allogeneic T lymphocytes and favored the proliferation of CD8+ cells. These data indicate that fever may cause a bias toward cellular responses, through its action on activated DCs, probably through CD40. Besides its physiological meaning, this phenomenon may have applications in immunotherapeutic strategies.