Descrições morfológicas são os primeiros passos para compreender a fisiologia dos organismos e seus sistemas. A biologia do desenvolvimento e a ontogenia da hematopoese de serpentes são pouco conhecidas, sendo a hematopoese embrionária inteiramente desconhecida. Os principais objetivos deste trabalho foram caracterizar o desenvolvimento e a hematopoese embrionária da serpente falsa-coral Oxyrhopus guibei (Dipsadidae: Xenodontinae) a partir da oviposição, através da análise de caracteres morfológicos externos para a classificação dos estágios de desenvolvimento embrionário, de 30 embriões com 1-3, 15, 25, 30, 45, 65 e ~75 dias de ovipostura (d.o.) (quando eclodem), e de cortes histológicos de 29 ovos (ou embriões) de 1-3, 15, 25, 30, 45, 60d.o. e de filhotes do dia da eclosão e com uma semana de nascimento para a caracterização da hematopoese embrionária. Os embriões de 1-3d.o. foram classificados entre os estágios (st.) 17 e 23, com ausência da formação ocular até a presença de olhos sem pigmentação; de 15d.o. no st. 26, com olhos pigmentados e ductos endolinfáticos calcificados; de 25d.o. no st. 31, com escamas no corpo não pigmentado e a musculatura dos flancos não fusionada na linha mediana ventral; de 30d.o. entre os st. 31 e 33, que em adição à formação descrita no período anterior, apresentam membrana palpebral completa e musculatura fusionadas na linha mediana ventral na região pré-cardíaca; de 45d.o. entre os st.34 e 36, com musculatura completamente fusionada na linha mediana ventral e começando a desenvolver um padrão de pigmentação; de 65d.o. no st. 37 (último da tabela) com o padrão de pigmentação bem desenvolvido de coral em tríades, típico da espécie, e com o dente do ovo, e com ~75d.o. os ovos eclodiram com filhotes sadios, que apresentam cicatriz umbilical e o dente do ovo permanece presente até dois dias após a eclosão. A hematopoese em ovos de 1-3 e 15d.o. foi caracterizadas como hematopoese extraembrionária, ocorrendo nos vasos das membranas extraembrionárias e na fenda vitelínica, e hematopoese intraembrionária, na região AGM (aorta gonadal mesonéfrons), principalmente no interior da artéria aorta dorsal, que continua como local hematopoético embrionário até no período de 30d.o., porém com estruturas mais diferenciadas; com 45d.o. o principal local hematopoético passa a ser medula óssea, com foco hematopoético de multi-linhagem sanguínea a partir de 60d.o., nos seios vertebrais e medula costal, constituindo o local hematopoético definitivo. Foi observada a atividade hematopoética junto ao tecido renal, com o desenvolvimento da região AGM, entre os períodos de 15 a 30d.o., o timo e baço apresentam diferenciação linfocítica, observados a partir de 30 e 45d.o., respectivamente e o fígado não apresenta hematopoese embrionária. Esta é a primeira descrição do desenvolvimento embrionário de uma serpente Caenophidia ovípara e da hematopoese embrionária de Serpentes, contribuindo para o conhecimento da fisilogia destes processos e indicando a necessidade de estudos tanto para um melhor entendimento do desenvolvimento embrionário, quanto para a compreensão da ontogenia da hematopoese das serpentes. / Morphological descriptions are the first steps to understand the physiology of organisms and their systems. Developmental biology and ontogeny of hematopoiesis of snakes are poorly known, and the embryonic hematopoiesis is completely unknown. The main goals of this study were to characterize the embryonic and hematopoiesis development of the false coral snake Oxyrhopus guibei (Dipsadidae: Xenodontinae) since oviposition. Analysis of external morphological characters regarding the classification developmental stages for developmental stages classification has been made for 30 embryos with 1 -3, 15, 25, 30, 45, 65 and ~ 75 days after oviposition days (o.d.) (when hatched), and histological sections of 29 eggs (or embryos) from 1-3, 15, 25, 30, 45, 60o.d. and one day neonates and one week after birth for the characterization of embryonic hematopoiesis. The embryos with 1-3o.d. were ranked between 17 and 23 stages (st.), with the absence of eye formation to the presence of unpigmented eyes; with 15o.d. in 26 st., with pigmented eyes and calcified endolymphatic duct; with 25o.d. in 31 st., with unpigmented body scales and muscles of the flanks not fused in the midline, with 30o.d. between 31 and 33 st., which in addition to the preceeding form, have full eyelid membrane and muscles fused at midline in the pre-cardiac region; with 45d.o. between 34 and 36 st., with muscles completely fused in the midline and beginning the development of pigmentation pattern, with 65o.d. in 37 st. (last stage of table) with the distinctive pigmentation tricolor-triad pattern , and the egg tooth, and ~ 75 d.o. health newborns have hatched, exhibiting the umbilical scar and the egg tooth up to two days after hatching. The hematopoiesis in eggs of 1-3 and 15o.d. was characterized as extraembryonic hematopoiesis, occurring in the vessels of extraembryonic membranes and in yolk clef, and intraembryonic hematopoiesis, in the AGM (gonad mesonefron aorta), mainly within the dorsal aorta, which continues as the embryonic hematopoietic site until the 30o.d., although with better differentiated structures; wth 45o.d. the bone marrow turns the main hematopoietic site, and with a hematopoietic focus of multi-lineage blood beginning in the 60d.o. at vertebral sinus and the ribs marrow, consisting the definitive hematopoietic sites. Hematopoietic activity was observed with the kidney tissue, through development of the AGM region, between the periods of 15 to 30d.o., thymus and spleen lymphocyte differentiation have been observed at 30 . and 45o.d., respectively, and the liver does not displays embryonic hematopoiesis. This is the first description of embryonic development of an oviparous Caenophidia snake and of the embryonic hematopoiesis in Serpentes, contributing to the knowledge the physiology of these processes and demonstrating the need of further studies for a better understanding of both embryonic development and the ontogeny snake.

A Ipomoea carnea é uma planta tóxica de distribuição nacional e encontrada também em outros países de clima tropical. Possui entre seus princípios ativos um alcalóide indolizidínico chamado suainsonina que, devido à sua ação sobre enzimas intracelulares (a &alpha;-manosidase ácida e a manosidase II do Complexo de Golgi), altera receptores de membrana e moléculas de adesão. Acredita-se que a suainsonina possa ter atividades imunomodulatória, mieloprotetora, antineoplásica e antimetastática. Devido ao potencial terapêutico sugerido pela literatura, nesse estudo avaliou-se os possíveis efeitos antineoplásico e mieloprotetor da suainsonina empregada isoladamente ou em associação com um antineoplásico clássico, a cisplatina. Além disso, procurou-se avaliar se a administração do resíduo aquoso final da Ipomoea carnea (RAF), isoladamente ou também associado à cisplatina, possui atividade anitineoplásica. Foram utilizados camundongos C57Bl/6, que foram inoculados por via intraperitoneal (ip.) com 2x107 células do tumor ascítico de Ehrlich. Os animais foram divididos nos grupos: BRANCO, CONTROLE e tratados com: cisplatina (CIS); cisplatina e suainsonina (CISSW); suainsonina (SW); cisplatina e RAF (CISRAF); e RAF (RAF). A suainsonina foi administrada por via ip. 2x ao dia (1mg/kg de peso vivo). A cisplatina foi administrada por via ip. em dias alternados (0,25mg/kg). A administração do RAF foi realizada por gavage, na dose de 3g de folhas secas/kg de I. carnea. Foi avaliada a sobrevida, o crescimento tumoral e a celularidade da medula óssea e do baço. Os resultados mostraram que apenas o grupo CISSW apresentou aumento significativo na sobrevida em relação ao grupo CONTROLE (Logrank test, p<0,05). Em relação ao crescimento tumoral, os grupos CIS e CISSW apresentaram diminuições significantes quanto comparados ao grupo CONTROLE; porém, o grupo CISSW apresentou melhores resultados (ANOVA com pós teste de Dunns, p<0,001) quando comparado ao grupo CONTROLE,enquanto os animais tratados com o RAF não apresentaram diferenças significantes. Dados sobre o ciclo celular (citometria de fluxo) revelaram que todos os animais tratados com a cisplatina (CIS, CISSW e CISRAF) apresentaram aumento significante no número de células tumorais em apoptose, porém o grupo CISSW apresentou melhores resultados, prejudicando de maneira mais eficiente o ciclo celular das células tumorais. As análises referentes à atividade mieloprotetora da suainsonina não apresentou dados relevantes. Estes dados em conjunto indicam efeito antineoplásico sinérgico da suainsonina quando administrada concomitantemente com a cisplatina. / Ipomoea carnea is a toxic plant widely distributed in Brazil and other tropical countries. Among its active principles I. carnea has the indolizidine alkaloid swainsonine, which inhibits intracellular enzymes (the lysosomal &alpha;-mannosidase and the Golgi mannosidase II) and alters membrane receptors and adhesion molecules. It is known that swainsonine presents the following activities: imunommodulatory, bone marrow protective, antineoplastic e antimetastatic. Due to the therapeutical potencial suggested by other authors, the present study was performed to evaluate the antineoplastic and bone marrow protective effects of swainsonine when used alone or in association with a classic chemotherapy agent like cisplatin. Moreover, the antineoplastic effect of the I. carnea aqueous fraction (AF), when administered alone or in association with cisplatin, was evaluated. For this, male C57Bl/6 mice were randomly assigned to seven groups: UNTREATED; CONTROL; CIS (treated with cisplatin); CIS-SW (treated with cisplatin and swainsonine); SW (treated with swainsonine); CIS-AF (treated with cisplatin and I. carnea aqueous fraction); and AF (treated only with I. carnea aqueous fraction). All animals except for those from the untreated group received an intraperitoneal (ip.) inoculation of 2x107 viable Ehrlich Ascites Carcinoma cells on day zero. Swainsonine was administered by the ip. route twice a day (1mg/kg of body weight), cisplatin was administered by the ip. route in alternate days (0.25mg/kg) and AF (3g of I. carnea dry leaves/kg) was administered by the oral route (gavage). Survival rate, tumor growth, bone marrow and spleen cellularity were evaluated. The only group that showed a significant increase on the survival rate when compared to CONTROL was CISSW (Logrank test, p<0.05). Considering tumor growth, both CIS e CISSW groups showed significant decreases in this parameter when compared to CONTROL. However, this decrease was better observed in the CISSW group (ANOVA followed by Dunns test, p<0,001), when compared to the CONTROL. Treatment with AF did not cause significant differences. The cell cycle results (flow citometry) revealed a significant increase in the number of apoptotic tumor cells in all animals treated with cisplatin (CIS, CISSW e CISRAF), although CISSW showed better results, altering the cycle of the tumor cells more efficiently. Swainsonine did not present any relevant protective activity on the bone marrow. These results suggest the existence of a synergism when swainsonine is administered in association with cisplatin.

Este trabalho aborda o uso da realidade virtual aplicada à simulação de procedimentos invasivos em oncologia pediátrica. Para tanto, apresenta uma revisão dos conceitos relacionados à concepção de simuladores baseados em realidade virtual, descrevendo requisitos específicos como estereoscopia e interação háptica. Particularmente, é apresentado um estudo de caso em coleta de medula óssea para transplante, para o qual um simulador foi desenvolvido. Este é o primeiro trabalho em simulação cirúrgica para oncologia pediátrica baseado em realidade virtual e apresenta detalhes relacionados à implementação do simulador e aspectos relacionados à calibragem de propriedades físicas em modelos tridimensionais. O trabalho também analisa e demonstra vantagens no uso de sistemas dessa natureza voltados para o treinamento médico, apontando necessidades e novos problemas a serem tratados por simuladores futuros. / This work approaches the use of the Virtual Reality applied to the simulation of invasive procedures in pediatric oncology. It presents a revision of the concepts related to the conception of simulators based on virtual reality, describing requirements as stereoscopy and haptic interaction. As specific problem, it was made a case study in bone marrow harvest for transplant, for which a simulator was developed. This is the first work in surgical simulation for pediatrics based on Virtual Reality and it presents details related to the implementation of the simulator and aspects related to physical properties in three-dimensional models. The work also analyzes and demonstrates advantages in the use of this kind of system for medical training besides to point needs and new problems to be addressed in future work.

A pessoa com distúrbios hematológicos demonstra desarranjos físicos, emocionais e sociais. Assim, atuar nesta área é estar disposta a cuidar do indivíduo, percebendo e intervindo precocemente nas alterações manifestadas. Os tratamentos para as patologias hematológicas podem ser feitos pela radioterapia, quimioterapia e o transplante de células tronco hematopoéticas (TCTH). Na maioria dos casos, a combinação destas modalidades terapêuticas faz-se necessária para o alcance do controle da doença. Após ser submetido à quimioterapia e/ou radioterapia o paciente é encaminhado para o TCTH almejando-se a cura. Estes tratamentos são altamente agressivos, acarretando prejuízos em todas as dimensões da vida do doente. A compreensão da trajetória da pessoa acometida por uma doença, desde o início dos sinais e sintomas até as perspectivas para o futuro, identificando suas idéias e condutas na luta pela sobrevivência, parte do entendimento do contexto sociocultural em que está inserida. Nessa perspectiva, o objetivo deste estudo foi identificar os modelos explicativos (MEs) para o TCTH alogênico aparentado, na visão de um grupo de pacientes. Para o seu alcance, estabelecemos como base teórica a antropologia médica, o método do estudo de caso qualitativo e a técnica de análise de conteúdo indutivo. Participaram deste estudo 11 pacientes, sendo três com leucemia mielóide aguda, quatro com leucemia mielóide crônica, dois com leucemia linfóide aguda e dois com anemia aplástica grave. As seis mulheres e os cinco homens formam um grupo de adultos em idade produtiva e suas características sociais mostram que são pessoas oriundas da classe social popular. Realizamos entrevistas semi-estruturadas, norteada por questões que integram a construção dos modelos explicativos. Posteriormente compilamos cada uma das entrevistas e seguimos para a análise de dados que se realizou em duas etapas. Reunimos as entrevistas e após várias leituras elencamos as categorias temáticas: da trajetória da doença ao tratamento especializado, os sentidos dados à doença e aos tratamentos, o lidar com a doença e os tratamentos e a vida após os tratamentos. Por estas categorias apreendemos os diversos elementos constitutivos dos modelos explicativos do adoecer e do submeter-se ao TCTH alogênico, onde evidenciamos a influência da cultura em que estão inseridos os participantes. Os entrevistados retrataram os sinais e sintomas da doença, a percepção do estar doente e suas causas, a difícil busca pela assistência à saúde, os tratamentos caseiros, a necessidade das terapêuticas, a existência de um doador compatível, as complicações e suas consequências, as dificuldades financeiras, os cuidados e as mudanças no modo de vida após os tratamentos e as perspectivas de futuro. Concluímos que a cultura é um sistema de referência para as pessoas de um grupo social, que fornece formas de pensar e agir sobre uma determinada situação ou evento. Evidenciamos exemplos da trajetória percorrida pelos participantes em busca da cura, almejando instigar os profissionais da saúde, em especial à equipe de enfermagem, que cuidam destes pacientes a olhá-los em todas as suas dimensões da vida, desejando um cuidado integral e diferenciado, integrando conhecimentos culturais ao modelo biomédico. / People with hematologic disorders face physical, emotional and social alterations. Thus, acting in this area means to be willing to care for the individual, precociously perceiving and intervening in the revealed changes. Hematologic pathologies can be treated by radiotherapy, chemotherapy and hematopoietic stem cell transplantation (HSCT). In most cases, the combination of these types of therapy is needed to control the disease. After undergoing chemotherapy and/or radiotherapy the patient is subject to HSCT, aiming the cure. These treatments are highly aggressive, causing damages in all dimensions of patients lives. Understanding patients sociocultural context is necessary to comprehend the trajectory of someone affected by a disease, since the beginning of the signs and symptoms up to the perspectives for the future, identifying their ideas and behaviors in the fight for survival. In this way, this study aimed to identify the explanatory models (EMs) for allogeneic HSCT of related donors, in the view of a group of patients. In order to achieve this, the medical anthropology, the qualitative case study method and the inductive content analysis technique were established as theoretical bases. Eleven patients participated in the study, three with acute myeloid leukemia, four with chronic myeloid leukemia, two with acute lymphoid leukemia and two with severe aplastic anemia. The six women and five men are adults at reproductive age and their social characteristics show they are from the lower social class. Semi-structured interviews guided by questions that are part of the construction of the explanatory models were carried. Afterwards the interviews were compiled and analyzed in a two-phase data analysis. Interviews were gathered and after several readings, the following thematic categories were listed: from the trajectory of the disease to specialized treatment, the meanings given to the disease and treatments, dealing with the disease and treatments, and life after treatments. Different elements of the explanatory models of becoming ill and undergoing allogeneic HSCT emerged from these categories. The influence of participants cultural context was evidenced. Interviewed subjects reported the signs and symptoms of the disease, the perception of being ill and its causes, the difficult search for health care, home treatments, the need of medications, the existence of a compatible donor, complications and their consequences, the financial difficulties, care and changes in the lifestyle after treatments and the perspectives of future. It is concluded culture is a reference system to people from a social group, which provides ways of thinking and acting about a certain situation or event. Examples of the trajectory followed by the participants in the search for care is evidenced, aiming to motivate health professionals, specially from the nursing team, who delivery care to these patients, to look them in all dimensions of their lives, to provide a comprehensive and differentiated care, integrating cultural knowledge to the biomedical model.

Orientador: Maria Aparecida Custódio Domingues / Resumo: Introdução: A medula óssea é o órgão responsável pela hematopoise, fenômeno que se relaciona com a diferenciação da célula tronco hemopoética (CTH) em eritrócitos, granulócitos, monócitos, linfócitos e plaquetas. É constituída pelo tecido hematopoiético, tecido gorduroso, matriz proteica e sinusoidais e está contida no interior dos ossos esponjosos, em contato com o tecido ósseo cortical e trabecular. Em 1978, Ray Schofield conceituou nicho como local especifico, dentro da medula óssea, onde a CTH se origina, assenta e prolifera. Atualmente sabe-se que o nicho é mais do que um simples compartimento morfológico, mas um sítio funcional e dinâmico, que se remodela e influencia a função da CTH e suas progenitoras, exercendo papel definitivo na CTH normal e neoplásica. Estudos demonstram que os nichos e seus componentes celulares e proteicos, influenciam e determinam a manutenção de CTH ativas e quiescentes, sendo determinante na manutenção, progressão e recaída de doenças da medula óssea. Objetivo: O objetivo deste trabalho foi avaliar na medula óssea humana, acometida por doenças primárias, secundárias e reacionais, o comportamento da CTH, descrevendo localização e quantificação destas. Material e métodos: Para realização deste estudo, foram selecionados quatro grupos de doenças hematológicas, divididas em mieloma múltiplo (n= 10), leucemia linfóide crônica (n= 10), síndrome mielodisplásica (n= 10) e reacional (n= 10). Foi realiado um estudo imuno-histoquímico para avaliação da ... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: Introduction: Bone marrow is the organ responsible for hematopoiesis, a phenomenon that is related to the differentiation of hemopoietic stem cell (HSC) into erythrocytes, granulocytes, monocytes, lymphocytes and platelets. It consists of hematopoietic tissue, fatty tissue, protein and sinusoidal matrix and is contained within the spongy bones, in contact with the cortical and trabecular bone tissue. In 1978, Ray Schofield conceptualized a niche as a specific site within the bone marrow, where the HSC originated, settled, and proliferated. It is now known that the niche is more than a simple morphological compartment, but a functional and dynamic site, which remodels and influences the function of HSC and its progenitors, playing a definitive role in normal and neoplastic HSC. Studies demonstrate that niches and their cellular and protein components influence and determine the maintenance of active and quiescent HSC, being determinant in the maintenance, progression and relapse of diseases of the bone marrow. Objective: The aim of this study was to evaluate the behavior of HSC in the human bone marrow, affected by primary, secondary and reactional diseases, describing their location and quantification. Materials and methods: Four groups of hematological diseases were selected, divided into multiple myeloma (n = 10), chronic lymphocytic leukemia (n = 10), myelodysplastic syndrome (n = 10) and reactional (n = 10). An immunohistochemical study was performed to evaluate the marki... (Complete abstract click electronic access below) / Mestre

The tumour microenvironment plays a key role in tumour progression. In this thesis acute myeloid leukaemia (AML) was used as a model system to investigate the interplay between stromal and cancer cells. AML is a heterogeneous clonal disorder of haematopoietic undifferentiated progenitor cells or 'blast cells', which accumulate in the bone marrow and lead to the reduced output of crucial haematopoietic elements. Due to its heterogeneity (at least in part), treatment of the disease has not witnessed great innovation in the past 30 years. The bone marrow microenvironment (BMM) has a key role in the haematological malignancies contributing to the survival of leukaemic blasts. Relapse in AML occurs because of residual disease and evidence suggests that this resistance is facilitated through leukaemic cells ability to reside in BMM niches. To understand the precise role of the BMM in AML progression and therefore target any supportive mechanisms requires knowledge of how AML cells communicate with their microenvironment. In the work presented in this thesis I undertook a multi-proteomic approach that utilised liquid chromatography tandem mass spectrometry (LC-MS/MS) to assess the interplay between AML and BMM cell signalling. This thesis shows the results of a secretomic analysis of stromal cell lines, which identified a previously uncharacterized panel of six stromal secreted proteins (BMP-1, CSF-1, CTGF, HGF, S100-A4 and S100-A11) that support primary AML cell survival and proliferation in culture. Comparison of AML cell signalling (using global phosphoproteomic methods) following treatment with the newly identified growth factors revealed that these signalling proteins elicit multi-nodular activation of signalling networks with known anti-apoptotic activity. Consistent with the cell signalling proteomics data, cell viability studies as a function of pharmacological kinase inhibitor treatment determined that the sensitivity of AML to targeted kinase inhibitors was modulated by the supportive stromal conditioned media. To investigate heterotypic signalling between cell populations, AML/stromal cell co-cultures were designed, tested and optimised. These studies identified additional activated pathways in AML cells that were only present when AML cells had physical interaction with stroma. Complementary analysis of the stromal cells which had been first cultured with AML cells revealed that despite heterogeneity there is an emerging stromal phospho-proteomic signature that is different in BMM independent AML cells vs BMM interactive AML cells. Collectively these findings evidence the influence that the BMM can have on AML signalling. Although evidence for the influence of BMM in modulating AML resistance to standard chemotherapy exists, this study highlights specific BMM components that contribute to the ability of AML cells to circumvent current treatments based on kinase targeted drugs. These observations have implications for designing future therapies for AML.

The mammalian target of rapamycin (mTOR) signalling pathway takes part in both extracellular and intracellular signals. It is a major regulator of cell metabolism, growth, proliferation and survival. mTOR also regulates critical processes such as cytoskeletal organization, ribosomal biogenesis, transcription and protein synthesis. The mTOR pathway has been implicated in many diseases such as cancer, neurodegeneration and diabetes, which impact homeostasis and cellular functions. Moreover, mTOR has also been shown to play a critical role in immune cell regulation of T and B cells together with neutrophils and antigen presenting cells, as it integrates signals between them extending to the entire immune microenvironment. The aim of my study was to investigate the role of a component of the mTOR complex 1, Raptor, in myeloid cells. My findings show that the absence of Raptor knock out (KO) does not affect bone marrow derived macrophage (BMDM) differentiation and maturation. However, the absence of Raptor influences BMDM polarisation towards an inflammatory phenotype, at least at the level of transcription as observed by increases in mRNA expression of inflammatory cytokines such as TNFα, IL-12β, and IL-6. This finding was consolidated by an increase in NFκΒ pathway signalling in Raptor KO BMDMs. Downstream intracellular signalling in myeloid cells was affected by deletion of Raptor as I found reduced S6K phosphorylation in Raptor KO BMDMs compared to wild type (WT) BMDMs. As a consequence of Raptor absence in BMDMs, STAT3 phosphorylation was also reduced. Raptor deletion did not impact the PI3K/Akt signalling pathway, but decreased phosphorylation of ERK. BMDMs lacking Raptor had reduced phagocytic activity as they were also observed to migrate less towards a pancreatic cancer cell line. However preliminary experiments in pancreatic cancer models did not indicate a major role for Raptor in the activity of tumour associated myeloid cells. My results demonstrate that Raptor and by implication mTORC1, is involved in macrophage polarisation and function.

Made available in DSpace on 2016-04-25T20:21:05Z (GMT). No. of bitstreams: 1 Maria de Fatima Patu da Silva.pdf: 2895041 bytes, checksum: a2389e9eedb7123cb1f8a76e5d10ad18 (MD5) Previous issue date: 2013-11-11 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / This work is a doctoral research in Social Sciences and whose object of study has been the bone marrow donor/transplantation. This can be analyzed at least by two areas of knowledge: Anthropology and Medicine. We take the notion of gift as a founding concept of our reflection. Thus, we should remember the bone marrow as an object donated for transplantation that carries beyond the genetic and physiological capacity, power and strength to revive the receiver. In recent decade, organ transplantation has attracted reviews from various sectors of the Brazilian society, the media, on the other hand, has been playing an important role in the broad disclosure information by democratizing the discussions, sometimes heated and contradictory, and clarifies or raises doubts and fear in people. It is therefore a matter of great social relevance and as such has been treated well by society; it has been receptive to the calls of solidarity in organ donation as public health, because it is a procedure of high complexity and high cost, requiring measures of public policy planning and resource allocation, democratic and ethical criteria. This study aims to describe the profile of recipients of bone marrow in Recife and Lisbon. We performed a retrospective analysis from 2002 to 2010. Over this period, 134 patients underwent transplantation with unrelated donors of these, 107 in Lisbon and 26 in Recife. Patients transplanted in Lisbon (51.4 %) are female; while in Recife (69.2 %) are male. Are in the age group 0-17 years in Portugal (54.2 %) and Recife (50 %) diagnosed with Aplastic Anemia prevalent. However, in Lisbon prevailed diagnosis of Acute Myeloid Leukemia. The study draws attention to the fact that Lisbon performed more transplants in the age group of 30-39 years old (18.7%), different from Recife (11.5%) in the same age group. In terms of age group 18-29 years old showed a percentage of (15.9 %). Research has shown acute myeloid leukemia as the most prevalent disease in Lisbon. It is a fact that confirms the literature, to affect individuals of all ages, especially those in adulthood. This study is in the age group above 50 years old in Lisbon (6.5%) and Recife (3.8 %). We emphasize that in the period of data collection, Recife had a population of 1,537,704 and 2,821,699 people Lisbon. Although Recife has other units, only the center where the research was conducted was doing this type of transplant / Este trabalho é uma pesquisa de doutorado em Ciências Sociais e tem como objeto de estudo o doador de medula óssea/transplante. Este pode ser analisado, pelo menos, por duas áreas do conhecimento: antropologia e medicina. Tomamos a noção de dádiva como conceito fundante da nossa reflexão. Para isso, convém lembrar que a medula óssea enquanto objeto doado para o transplante carrega além da constituição genética e sua capacidade fisiológica, o poder e a força de reviver o receptor. Nas últimas décadas, o transplante de órgãos tem atraído opiniões de setores diversificados da sociedade brasileira, a mídia, por sua vez, vem cumprindo importante papel na ampla divulgação ao democratizar as informações a discussões, algumas vezes acaloradas e contraditórias, assim como esclarece ou suscita dúvidas e receio nas pessoas. É, portanto, um tema de muita relevância social e como tal, tem sido tratado tanto pela sociedade, receptiva aos apelos da solidariedade na doação de órgãos, como pela saúde pública, por se tratar de um procedimento de alta complexidade e elevado custo, exigindo medidas de planejamento de políticas públicas e alocação de recursos, critérios éticos e democráticos. Este estudo objetiva descrever o perfil dos receptores de medula óssea em Recife e Lisboa. Foi realizada uma análise retrospectiva no período de 2002 a 2010. No período analisado, 134 pacientes realizaram transplante com doadores não aparentados destes, 107 em Lisboa e 26 em Recife. Os pacientes transplantados em Lisboa (51,4%) são do sexo feminino, enquanto em Recife (69,2%) pertence ao sexo masculino. Estão na faixa etária de 0 a 17 anos em Portugal (54,2%) e Recife (50 %) com diagnóstico prevalente de Anemia Aplástica. No entanto, em Lisboa prevaleceu o diagnóstico de Leucemia Mieloide Aguda. O estudo chama a atenção para o fato de que Lisboa realizou mais transplantes na faixa etária de 30 a 39 anos (18,7%), diferente de Recife (11,5%) na mesma faixa etária. Em relação à faixa etária de 18 a 29 anos, apresentou um percentual de (15,9%). A pesquisa mostrou a leucemia mieloide aguda como a doença mais prevalente em Lisboa. É um dado que corrobora com a literatura, por afetar indivíduos de todas as idades, principalmente os que estão na fase adulta. Nesse estudo está na faixa etária acima de 50 anos em Lisboa (6,5%) e Recife (3,8%). Ressaltamos que, no período da coleta de dados, Recife tinha uma população de 1.537.704 e Lisboa 2.821.699 pessoas. Apesar de Recife possuir outras unidades, apenas o centro onde foi realizada a pTesquisa estava fazendo este tipo de transplante

Les chimiokines (CKs) jouent un rôle important dans l’orchestration de la réponse immunitaireen contrôlant notamment la mobilisation des cellules immunitaires. Les cellules myéloïdes et, enparticulier, les monocytes sont impliquées dans le processus inflammatoire et notamment dansle développement des cancers. En effet, les cellules dérivées des monocytes ou macrophagesassociés aux tumeurs sont fortement représentés dans le micro environnement tumoral et sontsouvent associés à un mauvais pronostic. La caractérisation des mécanismes aboutissant aurecrutement des monocytes dans la tumeur représentent donc un enjeu majeur pourl’optimisation des protocoles thérapeutiques anti-cancéreux. Chez la souris, deux populationsmonocytaires sont caractérisées sur la base de l’expression différentielle des récepteurs auxchimiokines CCR2 et CX3CR1 et interviennent dans leur recrutement et leur différenciation enmacrophages dans les tissus, les monocytes inflammatoires (CCR2+, CX3CR1low) et les monocytesdits résidents (CCR2-, CX3CR1high). L’objectif principal de mon projet de recherche a été de mieuxcomprendre les mécanismes contrôlant la mobilisation des monocytes suite à un traitementchimiothérapeutique. J’ai entrepris d’étudier le rôle des récepteurs aux chimiokines, enparticulier CX3CR1, dans la reconstitution monocytaire après traitement chimiothérapeutiquepar le cyclophosphamide (CP), un agent alkylant reconnu pour son activité myélosuppressive. LeCP provoque un renouvellement des monocytes et une forte infiltration de la tumeur par les LTspécifiques de la tumeur (issus d’un transfert adoptif) associés à la réactivation de la réponseimmune anti-tumorale. Cependant, les LTs spécifiques de l’antigène de la tumeur se localisentpréférentiellement dans des zones riches en cellules dendritiques associées à la tumeur (TuDCs)et sont piégés par ces dernières. Ces interactions diminuent potentiellement le nombre decontacts entre les LT et les cellules tumorales suggérant un rôle pro tumoral des TuDCs. Letraitement au CP provoque une déplétion des cellules myéloïdes suivie d’une reconstitutionmassive des réservoirs de monocytes (moelle osseuse et rate). Au cours de la reconstitutionmonocytaire, l’expression de CX3CR1 diminue et est corrélée à une diminution de l’adhérence exvivo des cellules médullaires. Nous avons mis en évidence une mobilisation accrue desmonocytes inflammatoires au sein des souris CX3CR1- /- comparée aux souris WT et CCR2-/-.L’imagerie in vivo de la moelle osseuse au sein de souris CX3CR1-/- ou à l’aide d’un antagonistede CX3CR1 nous a permis de montrer un rôle spécifique de CX3CR1 dans le « crawling » sur lescellules endothéliales et le confinement des cellules monocytaires au niveau des sinus et duparenchyme médullaire. Nous suggérons qu’au cours de la mobilisation cellulaire induite par leCP le récepteur CX3CR1 contrôle la rétention médullaire des monocytes. Nous pensons que lamodulation du taux de mobilisation cellulaire au cours de la reconstitution induite par CP et/oule ciblage de CX3CR1 pourrait, par augmentation du pool de cellules myéloïdes leucocytairesd’un hôte, contribuer à l’amélioration des réponses cellulaires à la suite d’une lésion tissulaireou d’un dysfonctionnement des défenses immunitaires. De plus, le ciblage de CX3CR1 pourraittrouver des applications dans le domaine de la greffe de HSCs. / Chemokines orchestrate immune response especially by leucocytes mobilization. Myeloidcells, notably monocytes, are involved in inflammation and cancer development. Indeedmonocyte-derived cells and macrophages are strongly represented in tumourmicroenvironment and are associated with a bad prognosis. Characterization of mechanismsleading to monocytes recruitment within the tumor is thus a major issue in anti-cancertherapeutic protocols optimization. Based on the differential expression of chemokinereceptors CCR2 and CX3CR1, two populations of monocytes, inflammatory monocytes(CCR2+, CX3CR1low) and resident monocytes (CCR2-, CX3CR1high) have been characterized inmice which are involved in monocytes recruitment and differentiation into macrophages.The main objective of my work was to better understand the mechanisms of monocytesmobilization following chemotherapeutic treatment. I started to study the role of chemokinereceptors with a focus on CX3CR1 in monocytes reconstitution following cyclophosphamide(CP) treatment. CP is an alkylant agent known for its myelosuppressive properties. Thischemotherapeutic agent induces a transitory anti tumour immune response associated witha monocyte renewal and a strong infiltration of the tumour by adoptively transferred Tlymphocytes. However, antigen-specific T cells are trapped by tumour-associated dendriticcells (TuDCs). This potentially decreases interactions between T lymphocytes and tumourcells suggesting an immunosuppressive role of TuDCs. CP induces a strong depletion ofmyeloid cells followed by a massive reconstitution of bone marrow and spleen monocytesreservoirs. CX3CR1 expression on bone marrow monocytes is decreased duringreconstitution and correlated with a decreasing adhesion of these cells to CX3CL1 ex vivo.We highlighted an increased mobilization of inflammatory monocytes in CX3CR1-/- micecompared to WT and CCR2-/-. Intra vital imaging of bone marrow within CX3CR1-/- mice orwith the help of a CX3CR1 antagonist allowed us to show a specific role of CX3CR1 in thelumen crawling and confinement of monocyte-derived cells in both sinusoid andparenchyma of the bone marrow. We suggest that CX3CR1 controls the release of bonemarrow monocytes during CP-induced mobilization. We think that modulating the rate ofcellular mobilization, by increasing the host’s leukocyte pool during CP inducedreconstitution and/or targeting CX3CR1, could contribute to improve cellular responsefollowing tissue damage or immune cell dysfunction. Furthermore, targeting CX3CR1couldprovide applications in the stem cell transplantation domain.

Orientador: Maria Aparecida Custódio Domingues / Resumo: Introdução: A Leucemia Mieloide Crônica (LMC) consiste em uma neoplasia hematopoiética maligna de células-tronco, caracterizada pela aquisição do gene de fusão BCR-ABL1 associado à translocação [t(9;22) (q34;q11)]. O início é insidioso com progressão lenta seguida por uma fase acelerada, que culmina na crise blástica. Os nichos da medula óssea estão relacionados com o microambiente tumoral, afetando a sobrevivência e a proliferação das células-tronco leucêmicas. Objetivos: Análise da quantidade e distribuição de células-tronco hematopoiéticas nos diferentes nichos na LMC em diferentes fases e correlação destes achados com a distribuição de galectina-3 nas fases acelerada e blástica. Material e métodos: Revisão clínico-morfológica de 20 pacientes na fase crônica da LMC, 1 paciente na fase acelerada e 1 paciente na crise blástica. Análise imunofenotípica (CD34, CD31, CD44, CD117 e galectina-3) em Tissue Microarrays para avaliação dos diferentes nichos e sua correlação com a matriz proteica. Resultados: Observou-se aumento na expressão de todos os marcadores na fase crônica quando comparados ao controle. Na fase acelerada há aumento na expressão de CD44 e galectina-3 quando comparado ao controle enquanto que na crise blástica há aumento na expressão de CD44 e CD117. Conclusão: A LMC constitui uma neoplasia mieloproliferativa em que o remodelamento dos nichos malignos favorece a manutenção das células-tronco leucêmicas. O presente estudo demonstrou elevação em todos os marcadores... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: Introduction: Chronic Myeloid Leukemia (CML) is a malignant hematopoietic stem cell neoplasm, characterized by the acquisition of the translocation-associated BCR-ABL1 gene [t(9;22)(q34;q11)]. The onset is insidious with the slow progression followed by an accelerated phase, culminating in the blast crisis. The bone marrow niches are related to the tumor microenvironment, the survival and proliferation of the leukemic stem cells. Objectives: To analyze the amount and distribution of hematopoietic stem cells in different niches in CML in different phases and correlate these findings with the galectin-3 distribution in the accelerated and blastic phases. Material and methods: Clinical-morphological review of 20 patients in CML chronic phase, 1 patient in the accelerated phase and 1 patient in the blast crisis. Immunophenotypic analysis (CD34, CD31, CD44 and galectin-3) in Tissue Microarrays to evaluate the different niches and correlation with the protein matrix. Results: We observed the presence of all markers in the chronic phase when compared to the control. In the accelerated phase there is an increase in expression of CD44 and galectin-3, when compared to control while in blast crisis there is an increase in expression of CD44 and CD117. Conclusion: CML is a myeloproliferative neoplasm in which remodeling of malignant niches favors the maintenance of leukemic stem cells. The present study demonstrated an increase in expression of all immunohistochemical markers at differen... (Complete abstract click electronic access below) / Mestre