Submitted by Anderson Silva (avargas@icict.fiocruz.br) on 2012-10-22T16:59:52Z No. of bitstreams: 1 camila_s_lemos_ioc_bcm_0001_2010.pdf: 5583427 bytes, checksum: e0182171b95abb491ee7c70d18ffe4c1 (MD5) / Made available in DSpace on 2012-10-22T16:59:52Z (GMT). No. of bitstreams: 1 camila_s_lemos_ioc_bcm_0001_2010.pdf: 5583427 bytes, checksum: e0182171b95abb491ee7c70d18ffe4c1 (MD5) Previous issue date: 2010 / Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Rio de Janeiro, RJ, Brasil. / Para desvendar a fisiopatologia da leishmaniose humana, é necessário um melhor entendimento de como a Leishmania é capaz de sobreviver por anos em granulomas imunologicamente ativos. Neste estudo, utilizamos o modelo de infecção macaco Rhesus-Leishmania braziliensis (Macaca mulatta) de leishmaniose cutânea (LC), como um meio de avaliar a utilidade do sistema primata no estudo da resposta inflamatória crônica granulomatosa desenvolvida pelo hospedeiro tanto na forma auto-resolutiva quanto na forma persistente da doença. Nossos achados reforçam a noção de que existe uma interação entre o sistema imune do hospedeiro e a capacidade patogênica do parasita no resultado clínico da infecção por Leishmania. De acordo com os casos documentados de LC em humanos induzida por L.braziliensis, as infecções experimentais neste modelo induzem a célula “T-helper” tipo 1 (Th1) mediando-a inflamação e levando a formação de granulomas bem organizados, consistindo de todos os tipos celulares específicos encontrados nos granulomas humanos. Nós mostramos que os leucócitos polimorfonucleares são decisivos para controle do parasita nos eventos iniciais da inflamação in vivo. Ademais, fagócitos mononucleares que fagocitam granulócitos apoptóticos parasitados parecem ser a chave principal no estabelecimento da infecção nos macacos. A formação do granuloma verificada nos animais é direcionada por diversos mediadores inflamatórios que são importantes para o desenvolvimento das células Th1 e da função de macrófagos efetores. A análise cinética da resposta inflamatória revelou variados marcadores fenotípicos de células positivas para CD3, CD4, CD8, CD20, CD68, Foxp3 e HLA-DR. Digno de nota, agregados de células B CD20+ foram encontrados na lesão crônica, o qual serve como um substrato morfológico para orquestrarem ou modularem o desenvolvimento da resposta duradoura na infecção persistente. Nossos achados indicam que ambos os subgrupos de células regulatórias CD4 Foxp3+ e Foxp3- expressando interleucina-10 previne a erradicação da L. braziliensis da pele inflamada, desse modo, suprimindo uma eventual resposta curativa em macacos com LC persistente. Esses resultados apontam a viabilidade do uso desse modelo para elucidar, totalmente, os mecanismos moleculares das células t regulatórias mediadoras da supressão na resposta imune efetora, resultando assim o crescimento descontrolado do granuloma durante a leishmaniose, o que é necessário ser considerado em qualquer estratégia proposta para o controle terapêutico dessa importante doença inflamatória. / In order to unravel the physiopathology of leishmaniasis in humans, it is necessary to better understand how Leismania are able to survive for years within immunologicallyactive granulomas. In the present study, we used a Leishmania braziliensis-macaque (Macaca mulatta) infection model of cutaneous leishmaniasis (CL) as a means of assessing the usefulness of this primate system to study the host inflammatory granulomatous response involved in both healing and non-healing forms of the disease. Our findings reinforce the notion that an interplay exists between the host immune system and the pathogenic capability of the parasite in the clinical outcome of leishmanial infection. Consistent with documented cases of human L. braziliensis CL, experimental infections induced a T-helper cell type 1 (Th1)-mediated inflammation leading to the formation of well organized granulomas, consisting of all the specific cell types found within human granulomas. We provide evidence that polymorphonuclear neutrophil leukocytes are decisive for parasite containment by early inflammatory events in vivo. Furthermore, scavenger mononuclear phagocytes engulfing most of the parasite-laden apoptotic granulocytes are likely to be key players in establishing the infection in macaques. We show that granuloma formation in macaques is orchestrated by diverse inflammatory mediators that are important for Th1-cell development and macrophage effector functions. A kinetic analysis of the inflammatory response revealed marked variation in the number of positive cells for CD3, CD4, CD8, CD20, CD68, Foxp3, and HLA-DR phenotypes. Of note, CD20+ dense B-cell aggregates were found in chronic lesions, which might serve as a morphological substrate for the orchestration of the enduring host response to persistent infection. Our findings indicate that both Foxp3+ and Foxp3- CD4+ regulatory T-cell (Treg) subsets expressing interleukin-10 might prevent clearance of L. braziliensis from the inflamed skin, thereby eventually suppressing the healing response in macaques with long-standing CL. These results point to the feasibility of using this model to fully elucidate the molecular mechanisms of Treg-mediated suppression leading to the uncontrolled growth of granulomas occurring in leishmaniasis, which will need to be considered in any strategy designed for therapeutic control of this important inflammatory disease.

Nous possédons la faculté de reconnaître individuellement des centaines d’individus. Ceci nous permet d’évoluer dans une société complexe dont l’organisation est en partie forgée par les relations interindividuelles. La reconnaissance individuelle peut être réalisée par l'identification de divers éléments distincts, comme le visage ou la voix, qui forment chez l’Homme une seule représentation conceptuelle de l'identité de la personne. Nous avons démontré que les singes rhésus, comme les humains, reconnaissent individuellement leurs congénères familiers, mais également les individus humains connus. Ceci montre que la reconnaissance fine est une compétence partagée par un éventail d'espèces de primates pouvant servir de fondement à la vie en réseaux sociaux sophistiqués, et également que le cerveau s’adapte de façon flexible pour reconnaître les individus d'autres espèces lorsque ceux-ci ont une importance socioécologique. Par la suite, au niveau neuronal, ce projet a mis en lumière que les connaissances sociales concernant autrui sont représentées par les neurones hippocampiques ainsi que par les neurones inférotemporaux. Ainsi nous avons observé l’existence de neurones sélectifs aux visages non seulement dans le cortex inferotemporal, comme ceci a été décrit précédemment, mais également dans l’hippocampe. La comparaison des propriétés de ces neurones au sein de ces deux structures, suggère que les deux régions joueraient des rôles complémentaires au cours de la reconnaissance individuelle. Enfin, parce que l'hippocampe est une structure qui a évolué à des degrés divers chez différents mammifères pour soutenir la mémoire autobiographique et les représentations spatiales, la caractérisation des différents types de neurones et de leur connectivité a fourni un cadre commun pour comparer les fonctions de l’hippocampe à travers les espèces / Humans can individually recognize some hundreds of persons and therefore operate within a rich and complex society. Individual recognition can be achieved by identifying distinct elements such as the face or voice as belonging to one individual. In humans, those different cues are linked into one conceptual representation of individual identity. I demonstrated that rhesus monkeys, like humans, recognize familiarpeers but also familiar humans individually and that they match their voice to their corresponding memorized face. Thus it shows that fine individual recognition is a skill shared across a range of primate species, which may serve as the basis of a sophisticated social network. It also suggests that animals’ brains flexibly adapt to recognize individuals of other species when socio-ecologically relevant. Following at the neuronal level, this project put in light that social knowledge about other individuals is represented by hippocampal neurons as well as by inferotemporal neurons. For instance I observed the existence of face preferring neurons not only in the inferotemporal cortex as previously described but also in the hippocampus. Comparison of their properties across both structures, suggests that they could play complementary roles in recognition of individuals. Finally, because the hippocampus is a structure that evolved in various degrees to support autobiographical memory and spatial information in different mammals, I characterized the different subtypes of neurons and their network connectivity in the monkey hippocampus to provide a common anatomical framework to discuss hippocampal functions across species

Natal dispersal may have considerable social, ecological and evolutionary consequences. While speciesspecific dispersal strategies have received much attention, individual variation in dispersal decisions and its fitness consequences remain poorly understood. We investigated causes and consequences of natal dispersal age in rhesus macaques (Macaca mulatta), a species with male dispersal. Using long-term demographic and genetic data from a semi-free ranging population on Cayo Santiago, Puerto Rico, we analysed how the social environment such as maternal family, group and population characteristics affected the age at which males leave their natal group. While natal dispersal age was unrelated to most measures of group or population structure, our study confirmed earlier findings that sons of high-ranking mothers dispersed later than sons of low-ranking ones. Natal dispersal age did not affect males\\\'' subsequent survival, but males dispersing later were more likely to reproduce. Late dispersers were likely to start reproducing while still residing in their natal group, frequently produced extra-group offspring before natal dispersal and subsequently dispersed to the group in which they had fathered offspring more likely than expected. Hence, the timing of natal dispersal was affected by maternal rank and influenced male reproduction, which, in turn affected which group males dispersed to.

Magister Scientiae (Medical Bioscience) - MSc(MBS) / The incidence of male infertility is increasing, with up to 50% of infertile males having “unexplained” (idiopathic) infertility. Newly developed molecular techniques have great value in detecting subtle causes of male infertility, as compared to idiopathic infertility which may be explained by standardizing and optimizing sperm functional and structural tests in non-human primate (NHP) sperm. The aim of the study was to evaluate sperm functionality utilizing the sperm of two NHP species, i.e.1) the rhesus monkey (Macaca mulatta) and 2) the vervet monkey (Chlorocebus aethiops), and further evaluate the effect of physiological media (including commonly used, and newly formulated sperm wash and sperm capacitating media) on NHP sperm functionality. Sperm functionality was evaluated by investigating the following sperm functions i.e.: sperm motility, vitality, acrosome reaction (AR), hyperactivation, and mitochondrial membrane potential (MMP). Sperm functional tests included computer-aided semen analysis (CASA), motility analysis, BrightVit staining for sperm vitality, flourescenin isothiocyanate (FITC)- conjugated peanut agglutinin (PNA) staining for sperm acrosome integrity, induction of hyperactivation by stimulants (sperm preparation media containing capacitating ingredients), and mitochondrial inhibitor (Oligomycin-A) for testing MMP. All functional and structural tests were investigated in both species, except for acrosome integrity, mitochondrial inhibition and functional tests compared over time that could not be successfully completed and investigated in the rhesus species. Motility analysis tests proved that within the vervet species, the use of different physiological media results in statistically significant differences in motility and kinematic parameters over a 1 hour time period. Hyperactivation tests proved that capacitating physiological media produced significantly higher percentages hyperactivation when compared to sperm wash media within the vervet species over a 1 hour time period. Furthermore, within both NHP species, sperm structural analysis (vitality and acrosome integrity) results showed that no significant differences are present when making use of different physiological media over a period of 1 hour incubation. The incubation of vervet sperm with different concentrations of mitochondrial inhibitor, Oligomycin-A (0 μM, 5 μM, and 25 μM), resulted in motility inhibition over a 1 hour incubation period. By the evaluation of these tests it was found that the use of different sperm wash [Human tubal fluid (HTF), Ham‟s F-10® and HD Sperm Wash Plus (HDSWP)] and sperm capacitation media [Human tubal fluid with added caffeine (HTFC) and HD Sperm Capacitating Plus (HDSCP)] resulted in significantly different results within sperm functional tests as compared to sperm structural tests. The study indicates that the composition of media, varying from simple to more complex, used for semen preparation plays an important role in determining NHP sperm functionality. Based on these findings further investigation in larger NHP sample groups and human sperm are required to evaluate the role of certain ingredients in the development of more cost-effective media producing satisfactory results in terms of sperm functionality for artificial reproductive technologies (ART).

It has been hypothesized that head-unrestrained gaze shifts are controlled by an error signal produced by a feedback loop. It has also been hypothesized that the superior colliculus (SC) is within this feedback loop. If the feedback-to-SC hypothesis is valid, an unexpected mid-flight perturbation in gaze trajectory should be quickly followed by a concurrent change in the discharges of collicular saccade-related neurons. To verify this prediction experimentally, primate head movements were unexpectedly and briefly halted during head-unrestrained gaze shifts in the dark. Perturbed gaze shifts were composed of first a gaze saccade made when the head was immobilized by the head-brake, followed by a period where gaze was immobile, called a gaze plateau. The latter was composed of an initial period when the eyes and head were immobile, followed by a period wherein the head was released and the eyes counter-rotated to stabilize gaze. The plateau ended with a corrective gaze saccade to the goal location. In perturbed gaze shifts, there was widely distributed activity on the SC map during gaze plateaus, and there was no evidence that the initial motor program was aborted; the corrective gaze saccades were not "fresh" small stand-alone movements. Cells on the SC map responded at short latencies to head accelerations and associated gaze shift perturbations and carried a gaze position error (GPE = final - instantaneous gaze position) signal. As a large gaze shift progressed there was a caudo-rostral moving hill of activity on the SC map that encoded, not instantaneous veridical GPE, but a filtered version of it (time constant 100ms). Recordings from both the motor map and the so-called "fixation zone" in the rostral SC during perturbed head-unrestrained gaze shifts reveal gaze feedback control and a gaze feedback signal to the SC. However, these results do not prove that the SC is within the online gaze feedback loop, only that such a loop exists and that the collicular map is informed about its calculations.

Thesis (M.S.)--University of Wisconsin-Madison, 1962. / Includes bibliographical references (leaves 22-23).

Thesis (Ph. D.)--University of Wisconsin--Madison, 1990. / Typescript. Vita. eContent provider-neutral record in process. Description based on print version record. Includes bibliographical references (leaves 107-128).

Thesis (Ph. D.)--University of Washington, 2006. / Vita. Includes bibliographical references (p. 100-106).

Malaria is a pernicious disease that has greatly impacted and continues to affect the human population. While much research has been performed to understand the underlying nature of this disease, gaps in the knowledge-base persist. In order to address these deficiencies, a multi-disciplinary, multi-institutional project has been funded to study the systems biology of the host pathogen interaction during malaria infection in both humans and non-human primates. In the course of investigating the transcriptome during two 100-day experiments in Macaca mulatta, this work elucidated many of the underlying molecular pathways of the host and parasite that are affected by antimalarial drugs, as well as through host-pathogen interactions. The malaria-disease-related host pathways are related to, not surprisingly, immune-associated signalling and hematopoesis, and the altered parasite pathways demonstrate an association between disease severity and parasite life stage abundance. Continuing integration of this research with other data-types collected during the course of these experiments will improve our understanding of malaria systems biology and improve targeted malaria therapies.

No description available.