A ventilação natural pode ser promovida por dois mecanismos: o denominado efeito chaminé e o efeito dos ventos, porém também podem ocorrer os dois mecanismos simultaneamente. A determinação das vazões de ventilação por efeito do vento, em pavilhões industriais, pode ser realizada a partir de ensaios de modelos reduzidos em túnel de vento, com medição das velocidades do escoamento nas aberturas, ou então, com medição dos coeficientes de pressão em modelos fechados, nas faces onde estão localizadas as aberturas. A metodologia com medição dos coeficientes de pressão, que exige o emprego de um modelo teórico para calcular as vazões, é a forma convencional de usar o túnel de vento como ferramenta no projeto de ventilação. O objetivo deste trabalho é avaliar os procedimentos citados, para calcular as vazões de ventilação por efeito do vento em pavilhões industriais. Para atingir esse objetivo foram ensaiados em túnel de vento dois modelos reduzidos (escala geométrica 1:200) de um pavilhão industrial: um sem aberturas, para a medição dos coeficientes de pressão, e outro, com aberturas de ventilação de área variável, para a medição das velocidades do vento com anemômetro de fio quente. Além da comparação das duas metodologias para determinação das vazões de ventilação, os resultados dos ensaios foram também utilizados para analisar as alterações nas vazões de ventilação e nas velocidades do escoamento do ar decorrentes do fechamento de algumas aberturas do modelo estudado. Conclui-se que as aberturas localizadas na cumeeira e o tipo de escoamento exercem grande influência nas vazões de ventilação. A utilização da metodologia de medição direta de velocidades através de anemômetro de fio quente permite a obtenção de vazões de ventilação sem iterações, com resultados compatíveis com os modelos teóricos que se baseiam na diferença de pressões externas e internas. / Natural ventilation in industrial buildings may be promoted by two mechanisms: one due to wind effects and other called chimney effect. However, the two mechanisms may also occur simultaneously. The determination of wind induced ventilation flows in industrial buildings may be performed through wind tunnel tests, with measurement of discharge velocity at openings, or else, with measurement of the pressure coefficients at the face of the buildings, where the openings are located. The methodology which employs the pressure coefficients requires the use of a theoretical model to calculate the flows, being the conventional way of using the wind tunnel as a tool in the ventilation project. The objective of this work is to evaluate the experimental procedures previously mentioned in order to be able to calculate the wind induced ventilation flows in industrial buildings. To reach this objective, two reduced models of an industrial building were tested at a geometric scale 1:200; one without openings, for the measurement of the pressure coefficients, and the other with ventilation openings with variable area, for the measurement of the wind velocities by the use of hot-wire anemometer. Different opening combinations were also tested and the flows analyzed. It is concluded that the opening types and the different flows exert a big influence in the ventilation flows and, therefore, in the natural ventilation itself. Also, the direct velocity measurement technique, through hot-wire anemometer, allows the ventilation flows to obtained directly, without interactions, being the results compatible with the theoretical models based on pressure differences.

Ce travail s’articule sur trois axes ; le premier est une contribution à l'étude du répertoire des bactéries associées aux arthropodes vecteurs (tique et puces) en Afrique du nord (Algérie) et en Afrique Sub-saharienne (Bénin, Tanzanie et République Démocratique du Congo). Nous avons pu ainsi détecter par biologie moléculaire (qPCRs, PCR standard et séquençage) et pour la première fois au Bénin, Rickettsia typhi (l'agent du typhus murin), et Bartonella sp dans des puces collectées sur des rongeurs à Cotonou. Dans ce travail, nous avons également détecté Yersinia pestis, l'agent de la peste et R. felis (responsable de la fièvre boutonneuse) dans des puces de la RD du Congo. En Tanzanie, nous avons mis en évidence la présence de R. felis et R. typhi dans des puces de rongeurs. En Algérie, nous avons décrit pour la première fois la présence d'agent de borréliose de Lyme (Borrelia garinii) dans les tiques. Nous avons confirmé la présence de R. massiliae, R. monacensis R. aeschlimannii, R. slovaca et R. felis et nous avons également détecté pour la première fois en Algérie, Bartonella tamiae, une bactérie dont la pathogénicité est peu connue et Coxiella burnetii, l'agent de la fièvre Q.Dans la deuxième partie de notre travail, nous nous sommes intéressés à l’évaluation des compétences vectorielles des puces de chat (Ctenocephalides felis) et punaises de lit (Cimex lectularius) dans la transmission de l’agent de la fièvre des tranchées (Bartonella quintana) dont le vecteur connu est le pou de corps. Trois approches ont été utilisées : la qPCR, la culture et l’immunohistochimie. / This work focuses on three areas; the first is a contribution to the study of the repertoire of bacteria associated with arthropod vectors (tick and flea) in North Africa (Algeria) and in Sub-Saharan Africa (Benin, Tanzania and the Democratic Republic of Congo). We could thus detected by molecular tools (qPCRs, standard PCR and sequencing) and for the first time in Benin, Rickettsia typhi (the agent of murine typhus) and Bartonella sp in fleas collected from rodents in Cotonou. In this work, we have also associated the agent of plague (Yersinia pestis), and for the first time in fleas of DR of Congo, and we detected also R. felis (the causative agent of spotted fever). In Tanzania, we have highlighted the presence of R. typhi and R. felis fleas on rodents. In Algeria, we described for the first time the presence of Lyme disease agent (Borrelia garinii) in hard ticks. We confirmed the presence of R. massiliae, R. monacensis, R. aeschlimannii, R. slovaca and R. felis, we also detected for the first time Bartonella tamiae and Coxiella burnetii associated with bat ticks in Algeria.Regarding the second part we was interested in the evaluation of vector competence of cat fleas (Ctenocephalides felis) and bed bugs (Cimex lectularius) in the transmission of trench fever agent (Bartonella quintana) that is known to be transmitted by lice. Three approaches have been tested; qPCR, culture and immunohistochemistry.

Orientador: Marco Antonio de Andrade Belo / Resumo: O presente estudo investigou os efeitos da ciclofosfamida (CYP) na modulação de proteínas de fase aguda (APPs) e acúmulo celular em exsudatos presentes na bexiga natatória de tilápias (modelo de peixe de aerocistite) e o recrutamento de leucócitos na lamínula de vidro implantadas no tecido subcutâneo. Foram utilizadas 140 tilápias (Oreochromis niloticus) (± 150 g) distribuídas em 14 aquários com capacidade para 250 L de água (n = 10). Os animais foram distribuídos em dois ensaios: Experimento I: foram utilizados 80 animais distribuídos em quatro tratamentos: T1 - controle NAIVE; T2 - controle negativo + solução salina injetada NaCl; T3- controle positivo + injetado com Aeromonas hydrophila; T4- ciclofosfamida, Isopac ® Sigma (200 mg / kg, dose única, por via intraperitoneal) + injetado com A. hydrophila. As amostras de exsudato e sangue foram coletadas 6 e 24 horas após a infecção (HPI) para a contagem de células e determinação do fracionamento eletroforético das APPs por SDS-PAGE, digestão de proteínas em gel e identificação por espectrometria de massa. Experimento II: os peixes foram alocados (n=60) em 3 grupos experimentais: G1 - controle NAIVE; G2 – controle positivo + implante de lamínula; G3 - ciclofosfamida, Isopac ® Sigma (200 mg / kg, dose única, por via intraperitoneal) + implante de lamínula. Após serem anestesiados, foram submetidos ao implante da lamínula de vidro de 10mm de diâmetro no tecido subcutâneo. As amostras de sangue foram coletadas e as lamínulas foram... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: The present study investigated the effects of cyclophosphamide (CYP) on the modulation of acute phase proteins (APPs) and cellular accumulation in exudates present in the swim bladder of tilapia (aerocystitis fish model) and the recruitment of leukocytes on a glass cover slip implanted in the subcutaneous tissue. One hundred forty tilapia (Oreochromis niloticus) (± 150 g) were distributed in 14 aquariums with capacity of 250 L (n = 10), for two trials. Experiment I: For this purpose, 80 fish were distributed in four treatments: T1 – naïve control; T2 - negative control + injected with NaCl solution; T3- positive control + injected with A. hydrophila; and T4- cyclophosphamide (200 mg/kg) + injected with A. hydrophila. Samples of exudate and blood were collected 6 and 24 hours post-inoculation (HPI) for cell counts and determination of the APP electrophoretic fractionation by SDS-PAGE, in-gel protein digestion and mass spectrometric identification. Experiment II: Fish were allocated (n = 60) in 3 experimental groups: G1 – naive control; G2 - positive control + coverlet implant; and G3 - cyclophosphamide (200 mg/kg, single dose intraperitoneally) + cover slip implant. The glass coverslips (10 mm diameter) were implanted after anesthetization in the subcutaneous tissue. Blood samples were collected and the coverslips were carefully removed and washed with 0.9% saline 74 and 144 hours implantation. Then they were fixed in Bouin's solution and stained with hematoxylineosin. The tot... (Complete abstract click electronic access below) / Doutor

Colorectal carcinoma is an important, feasible and attractive target for chemoprevention because a) it is a major cause of mortality in the United States and in other developed countries worldwide, b) there is a high mortality associated with advanced disease, c) there is a well described molecular carcinogenesis pathway and d) recent advances in molecular genetics will improve the ability to identify high-risk subjects. Epidemiological data, colonoscopic screening and advances in molecular genetics has made possible the identification and selection of subjects at increased risk of developing colorectal cancer. Due to this new information it may be possible to impede malignant cellular transformation with drugs. Such intervention with relatively simple maneuvers, such as a low daily dose of aspirin, can potentially reduce mortality from colorectal cancer. Prospective trials need to confirm experimental and epidemiological data supporting the efficacy of aspirin and other NSAID as chemopreventive agents before they can be used in the general population at risk. To use cancer chemopreventives effectively and safely in an asymptomatic population, the risks should be minimized and the benefits maximized by determination of optimal dose, schedule and chemopreventive mechanism of the NSAID. By linking the putative mechanism of drug action to effect endpoints, we expect to know whether the chemopreventive intervention is likely to be effective in a given individual.

Neosporosis is a parasite disease caused by Neospora caninum protozoan and characterized as a major cause of abortion in cattle. Eventually is possible birth of animals with neurological signs, stillbirths or birth of clinically normal calves, but chronically infected. Although there are many researches related to bovine neosporosis, there are still doubts about the cycle and the pathogenesis of N. caninum infection, so that is suitable for certain control and prophylaxis strategies to disease. Gerbils are considered highly sensitive of the parasite replication. Thus, this study aimed to verify the occurrence of infection with N. caninum by oral inoculation of tachyzoites in gerbils (M. unguiculatus). Seventeen animals were orally inoculated with 4x105 N. caninum tachyzoites. The detection frequency of total DNA samples of N. caninum showed that the protozoan infectivity occurs via this route of inoculation, noting that 81.8 % of the inoculated animal parasite DNA was detected. In relation to tissues, it can be observed that the DNA parasite was found in all organs evaluated (heart, lungs, kidneys, liver, spleen, brain), by variable frequency, demonstrating the wide distribution of this parasite in animals tissues. / Neosporose é uma enfermidade de origem parasitária, causada pelo protozoário Neospora caninum e caracteriza-se como uma das principais causas de aborto em bovinos. Eventualmente, é possível ocorrer o nascimento de animais com sinais neurológicos, natimortos, ou ainda, o nascimento de bezerros clinicamente normais, porém cronicamente infectados. Embora existam inúmeras pesquisas relacionadas à neosporose bovina, ainda existem imprecisões quanto ao ciclo e patogenia da infecção por N. caninum, para que possam ser determinadas estratégias adequadas ao controle e profilaxia da doença. Os gerbils (Meriones unguiculatus) são considerados altamente sensíveis a replicação do protozoário. Assim, este estudo teve como principal objetivo verificar a ocorrência de infecção pelo N. caninum através da inoculação oral de taquizoítos em gerbils. Foram inoculados 17 animais pela via oral, com 4x105 taquizoítos de N. caninum. A frequência de detecção total de DNA de N. caninum das amostras demonstrou que ocorre infectividade pelo protozoário através desta via de inoculação, observando que em 81,8% dos animais inoculados foi detectado DNA do parasita. Em relação aos tecidos, pode-se observar que o DNA do protozoário foi encontrado em todos os órgãos avaliados (coração, pulmões, rins, fígado, baço, cérebro), com frequências variáveis, demonstrando assim a ampla distribuição deste protozoário em diferentes tecidos dos animais.

Submitted by Luciana Ferreira (lucgeral@gmail.com) on 2015-11-20T10:54:01Z No. of bitstreams: 2 Dissertação - Dieferson da Costa Estrela - 2015.pdf: 2754827 bytes, checksum: 254f2e157991239ed42ccec84b03e2dd (MD5) license_rdf: 23148 bytes, checksum: 9da0b6dfac957114c6a7714714b86306 (MD5) / Approved for entry into archive by Luciana Ferreira (lucgeral@gmail.com) on 2015-11-20T10:56:04Z (GMT) No. of bitstreams: 2 Dissertação - Dieferson da Costa Estrela - 2015.pdf: 2754827 bytes, checksum: 254f2e157991239ed42ccec84b03e2dd (MD5) license_rdf: 23148 bytes, checksum: 9da0b6dfac957114c6a7714714b86306 (MD5) / Made available in DSpace on 2015-11-20T10:56:04Z (GMT). No. of bitstreams: 2 Dissertação - Dieferson da Costa Estrela - 2015.pdf: 2754827 bytes, checksum: 254f2e157991239ed42ccec84b03e2dd (MD5) license_rdf: 23148 bytes, checksum: 9da0b6dfac957114c6a7714714b86306 (MD5) Previous issue date: 2015-08-17 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES / Obesity and chronic stress have been considered major public health problems that affect millions of people worldwide. The objective of this study was to analyze the effect of obesity associated with chronic stress on neurobehavioral parameters, hematological and biochemical in rats, considering that the association of these diseases can enhance their negative effects on homeostasis of the organism. For this, female Wistar rats were distributed in the following groups: standard diet (SD), standard diet + stress (DP + stress), cafeteria diet (Cafe) and cafeteria diet + stress (Cafe + stress) groups (n=12 rats, per group). The animals of the DP and DP + stress groups were fed standard food. Groups Cafe and Cafe + stress, additionally to the standard feed, were offered palatable and calorie-rich processed food, water with sucrose (300 g/L)and cola-type soft drink ad libitum. From the eighth experimental week, groups Std + stress and Cafe + stress were subjected to restraint chronic stress model (50 days). After the restraint chronic stress the animals were subjected to predictive anxiety tests (open field and elevated plus maze tests) and depression (forced swimming test). At the end of the experiment was evaluated hematological parameters (erythrogram, leucogram and platelet parameters) and biochemical (lipid profile, blood glucose, total proteins and fractions). The results demonstrate that cafeteria diet was able to induce obesity in the above animals, increasing body mass, weight of retroperitoneal and visceral fat, body mass index, specific rate of weight gain, weight delta efficacy coefficient food, weight gain coefficient for calorie intake, nasoanal length, abdominal circumference and kilocalories ingested. The ratio locomotion in the central quadrants/total locomotion evaluated during the open field test was not indicative of anxiogenic or anxiolytic effect in the animal’s behavior. However, the elevated plus maze test showed that obese and stressed animals were prone to higher anxiety levels. In addition, the obese and stressed animals display less climbing behavior than all the other groups, which can be considered an indicator of depression-like behavior. No damaging effects caused by the interaction of obesity and stress were observed on the biochemical and hematological parameters. Only individual effects of the factor obesity on leukocyte and glycemic parameters were observed. The obese animals presented hypoglycemia and leukocytosis by neutrophilia and lymphocytosis. / A obesidade e estresse crônico têm sido considerados importantes problemas de saúde pública que afetam milhões de pessoas em todo o mundo. Assim, o objetivo deste estudo foi analisar o efeito da obesidade associada ao estresse crônico sobre os parâmetros neurocomportamentais, hematológicos e bioquímicos em ratas, considerando que a associação dessas doenças pode potencializar seus efeitos negativos sobre a homeostase no organismo. Para isso, ratas Wistar foram distribuídas nos seguintes grupos experimentais: dieta padrão (DP), dieta padrão + estresse (DP + estresse), dieta de cafeteria (Cafe) e dieta de cafeteria + estresse (Cafe + estresse) grupos, com 12 animais por grupo. Os animais dos grupos e DP e DP + estresse foram alimentados com ração padrão roedoroes. Aos animais dos grupos Cafe e Cafe + estresse, adicionalmente à ração padrão, foram oferecidos diferentes alimentos palatáveis (calóricos e in natura), água com sacarose (300 g/L) e refrigerante do tipo cola, ad libitum. A partir da oitava semana experimental, os grupos DP + estresse e Cafe + estresse foram submetidos ao modelo de estresse crônico por restrição, durante 50 dias. Após o protocolo de estresse os animais foram submetidos a testes neucomportamentais preditivos de ansiedade (tesde do campo aberto e labirinto em cruz elevado) e depressão (teste do nado forçado). Ao final do experimento, avaliou-se parâmetros hematológicos (eritograma, leucograma e plaquetograma) e bioquímicos (lipidograma, glicemia, proteínas totais e frações). Os resultados demonstram que a dieta de cafeteria foi capaz de induzir um quadro de obesidade nos animais, aumentando a massa corpórea, massa das gorduras retroperitoneal e visceral, índice de massa corpórea, taxa específica de ganho de peso, delta do peso, coeficiente de eficácia alimentar, coeficiente de ganho de peso por consumo calórico, comprimento naso-anal, circunferência abdominal e quilocalorias ingeridas. A proporção da locomoção nos quadrantes centrais/locomoção total, avaliado durante o teste de campo aberto não indicou comportamento ansiolítico ou ansiogênico nos animais. No entanto, o teste de labirinto em cruz elevado mostrou que os animais obesos e estressados apresentaram maior índice de ansiedade. Além disso, os animais estressados e obesos exibiram menor comportamento de escalada em relação aos demais grupos, o que pode ser considerado um comportamento preditivo de depressão. Não foram observados efeitos prejudiciais da interação obesidade e estresse sobre os parâmetros bioquímicos e hematológicos avaliados. Evidenciaram-se apenas efeitos individuais do fator “nutrição” (fator 1) sobre os parâmetros leucocitários e glicêmicos avaliados. Os animais obesos apresentaram-se hiperglicêmicos e com quadro de leucocitose por neutrofilia e lifocitose.

Le tendon transmet les forces musculaires aux os et permet le mouvement. C'est un tissucomplexe et organisé, formé de cellules et d’une matrice extracellulaire (fibres collagènes,élastiques et substance fondamentale). Les fibres lui confèrent son élasticité et résistance et lasubstance fondamentale sa viscosité.La tendinopathie est souvent lié au sport, de morbidité élevée qui affecte laperformance des sportifs. Elle associe dégénérescence, douleur, rigidité et oedème local.L’imagerie quantifie et qualifie les changements morphologiques du tendon. L’homéostasietendineuse et sa physiopathologie restent peu connues, mais un changement des liaisons desfibrilles lors de la cicatrisation diminue sa résistance.L’objectif de l’étude est: mettre en place un modèle de tendinopathie chirurgicale chezla chèvre avec suivi clinique transposable (homme et cheval); développer un dispositifd’évaluation du comportement mécanique des tendons; et valider la chèvre comme modèleanimal pour l’étude de la tendinopathie. L’étude a été faite sur 84 jours avec un suivi cliniqueet échographique suivi d’un test de traction.La chèvre est un ongulé facile à héberger et manipuler. La technique chirurgicalepermet la résection de quelques fibres causant une lésion identifiable par échographie et dessymptômes cliniques. La grande variabilité du comportement mécanique ne se corrèle pas aupoids, morphologie des animaux ou section transversale du ses tendon, ce qui complique laréduction de l’effective. Notre modèle semble pouvoir être utilisé pour évaluer des traitementsde tendinopathies destinés à l’homme et/ou au cheval. Ce que n’exclue pas sondéveloppement pour renforcer sa pertinence / Tendons transmit muscles forces to bone and allow movement. It is a complexorganized tissue, composed by few cells and an extracellular matrix (collagen fibers, elasticfibers, and a ground substance. Fibers give to tendon its elastic characteristic and resistanceand ground substance its viscosity.Tendinopathy is a medical problem associated with sport, with a high morbidity whichaffects performance. It is an association of degeneration, pain, stiffness and local edema.Imaging can be used to quantify and qualify the changes in the morphology of the tendon.The tendon homeostasis and physiopathology remain poorly understood, but a change at theconnections of the fibrils, decreasing its strength.The purpose of this study is: development of surgical model of induced localtendinopathy in goats with clinical follow up transposable for humans and horses;development of a device to evaluate the mechanical properties of tendons, and demonstratethat the goat is a good experimental animal model for the study of tendinopathy. The studyhas been performed in 84 days with a clinical follow up, US imaging and tensile test.Goat is an ungulate easy to house and handle. The modified splitting allows easy andreproducible resection of some tendon fibers with an identifiable lesion in US imaging andclinical symptoms. The variability of mechanical behavior is not correlated with"macroscopic" aspects (weight, animal morphology or cross sectional area), thus reducing theeffective is difficult. However, our model appears in the state, can be used to evaluatetreatments for human and horse’s tendinopathy. The development of our model couldstrengthen its relevance.

Conselho Nacional de Desenvolvimento Científico e Tecnológico / Contagious ecthyma (orf) is a cutaneous disease that affects sheep and goats, and may be occasionally transmitted to humans. The disease is caused by orf virus (ORFV). ORFV infection produces croustous and proliferative lesions, usually on the nostrils and labial commissures of lambs, and also in the udder, teat skin and coronary bands of adults animals. The pathogenesis of ORFV infection is poorly understood and a search for an adequate animal model is required, yet the disease has been already reproduced in sheep, goats and rabbits. This dissertation relates the clinical, virological and pathological aspects of ORFV infection in rabbits and mice experimental inoculated. Ten rabbits, ten mice and two lambs were inoculated intradermally after skin scarification with an hypodermic needle. A viral suspension of ORFV IA-82 strain (108.5TCID50/mL) was inoculated in the internal face of the ear, back skin and labial commissure of rabbits; internal face of the ear of mice. Lambs were inoculated in the labial commissures and in the internal face of hind limbs. All animals were monitored clinically, virologically, and pathologically for 21 days. All rabbits developed clinical signs in the inoculation sites, begining with mild hyperemia that evolved to macules, papules, vesicle, pustules and scabs. Lesions appeared at days 3 and 4 post-inoculation (pi) and lasted to 3 to 10 days. Viral shedding was detected from days 2 to 14pi. Histological examination of lesions revealed focal proliferative dermatitis with ballooning degeneration and eosinophilic intracytoplasmic inclusions in keratinocytes, histological hallmarks of contagious ecthyma in sheep. A similar, albeit much milder clinical course was observed in 5 out of 10 inoculated mice. All lambs presented characteristic contagious ecthyma clinical and histopathologycal lesions from days 3 to 18pi, and the virus was recovered from lesions between days 2 and 19pi. At day 28pi, seroneutralization test (SN) was unable to detect neutralizing antibodies in all inoculated animals. These findings show that ORFV replicates and produce local lesions in rabbits and mice. However, rabbits are more susceptible to infection and disease, and may be used as an animal model to study some aspects of ORFV pathogenesis. / O ectima contagioso (ou orf) é uma doença infecto-contagiosa de pele que afeta principalmente ovinos e caprinos, e que ocasionalmente pode acometer o homem. O seu agente etiológico é o vírus da orf (ORFV). O ORFV produz lesões proliferativas, geralmente na comissura labial e no plano naso-labial de cordeiros, e também na pele do úbere, nos tetos e no rodete coronário dos cascos de animais adultos. A patogenia da infecção pelo ORFV é pouco conhecida, embora a doença já tenha sido reproduzida em ovinos, caprinos e coelhos. Essa dissertação relata os achados clínicos, virológicos e histopatológicos da infecção experimental de coelhos e camundongos pelo ORFV. Para isso, coelhos, camundongos e cordeiros foram inoculados pela via intradérmica (ID), após escarificação da pele com agulha hipodérmica. A inoculação dos cordeiros serviu como controle positivo. Uma suspensão viral da cepa IA-82 do ORFV (108,5DICC50/mL) foi inoculada na face interna da orelha, na pele do dorso e na comissura labial dos coelhos; na face interna da orelha dos camundongos; e na comissura labial e face interna do membro pélvico dos cordeiros. Os animais foram monitorados por 21 dias nos aspectos clínicos, virológicos e patológicos. Todos os coelhos inoculados apresentaram lesões semelhantes nos locais de inoculação, iniciando com hiperemia, evoluindo para máculas, pápulas, vesículas, pústulas e crostas. Os sinais surgiram 3 a 4 dias pós inoculação (pi) e duraram por 3 a 10 dias. Excreção viral foi detectada entre os dias 2 e 14pi. A análise histológica das lesões revelou dermatite focal proliferativa, com degeneração balonosa e corpúsculos de inclusão intracitoplasmáticos eosinofílicos nos queratinócitos, semelhante às alterações histológicas observadas nos cordeiros. Lesões similares, mas de menor intensidade foram observadas em 5 de 10 camundongos. Os cordeiros, utilizados como controles positivos, apresentaram lesões clínicas e histopatológicas características de ectima contagioso entre os dias 3 e 18pi, sendo que o vírus foi recuperado das lesões entre os dias 2 e 19dpi. No dia 28pi, pelo teste de soroneutralização (SN), não foram detectados anticorpos neutralizantes no soro dos animais inoculados. Esses resultados demonstram que a inoculação de ORFV resulta em replicação viral e produção de lesões em coelhos e camundongos, porém a doença é reproduzida de forma mais consistente em coelhos. Portanto, sugere-se que coelhos possam ser utilizados como modelos para estudos in vivo com o ORFV.

Metabolic syndrome (MetS) is a complex condition with a number of interacting genes, epigenetic and environmental factors underlying its pathogenesis. The analysis of genetic component of MetS showed that number of defining parameters of the syndrome is linked to regions of rat chromosome 4. In order to verify these quantitative trait loci (QTL), a double congenic strain was derived with parts of chromosome 4 of spontaneously hypertensive rat (SHR, an inbred MetS model) origin introgressed onto genomic background of congenic Brown Norway strain (BN-Lx). The aim of the proposed thesis is comprise detail genetic mapping of differential segments of the above mentioned double congenic strain BN-Lx.SHR4 and comparison of its metabolic profile under different dietary conditions with varying carbohydrate and fat content. Utilizing DNA sequence and gene expression comparisons, candidate genes or polymorphisms for the MetS aspects and potential nutrigenetic interactions will be identified. Key words: nutrigenetics, experimental models, metabolic syndrome, congenic strain, genotyping, rat

L’injection intraveineuse de l’activateur tissulaire du plasminogène (tPA) est le seul traitement aigu de l’ischémie cérébrale autorisé chez l’Homme. Cependant, la thrombolyse présente des limites d’utilisation, comme son étroite fenêtre thérapeutique, un risque hémorragique et une efficacité de recanalisation malgré tout relativement peu élevée. De plus, la littérature suggère fortement que non seulement le tPA endogène, mais aussi exogène (capable de traverser la barrière hémato-encéphalique), a des effets pro-excitotoxiques. Nous avons proposé que cet effet résulte du clivage de la sous unité NR1 du récepteur NMDA. Malgré un effort important de la communauté scientifique pour chercher de nouveaux traitements, tous les espoirs se sont avérés être des échecs. Sur ces bases, ces travaux de thèse ont consisté à : 1) Améliorer les approches précliniques en développant un nouveau modèle d’ischémie cérébrale chez la souris et en incluant dans les études un des principaux facteur de risque des AVC, le vieillissement ; 2) Développer une stratégie d’immunothérapie visant l’interaction tPA/ récepteur NMDA. J’ai ainsi montré qu’il existe une diminution du volume de lésion ischémique corrélée à l’âge et que cette diminution de tPA est due à une diminution d’expression du facteur de transcription D-Site Albumin Binding Protein (DBP). J’ai également développé un modèle innovant d’ischémie thrombo-embolique chez la souris, dans lequel la reperfusion par le tPA est bénéfique, si tant est qu’elle soit réalisée de manière précoce. Sur ce modèle, j’ai apporté par une stratégie d’immunisation active la preuve in vivo du clivage du domaine amino-terminal de la sous-unité NR1 des récepteurs NMDA. Enfin, j’ai produit un anticorps médicament, capable d’empêcher l’interaction du tPA avec la sous-unité NR1 des récepteurs NMDA, dont une injection unique permet de réduire les lésions ischémiques, mais aussi d’augmenter la fenêtre thérapeutique de la thrombolyse, conférant alors une récupération fonctionnelle à long terme. Cette stratégie pourrait donc accroître la proportion de patients traitables après un AVC ischémique aiguë / Reperfusion with tissue plasminogen activator (tPA) is the only approved treatment for ischemic stroke. However, thrombolysis has some limitations, including a narrow therapeutic window, an elevated risk of hemorrhage transformation and a low level of effective recanalization. Moreover, there is a growing body of evidence that both endogenous and exogenous tPA (able to cross the blood-brain barrier) could mediated pro-excitotoxic effects. We have proposed that this noxious effect results from the cleavage of the NR1 subunit of the NMDA receptor. My thesis work consisted in: 1) Improving pre-clinic approaches by developing a new model of thrombo-embolic ischemia in mice and by taking into account a major risk factor for stroke, aging; 2) Developing a strategy of immunotherapy targeting the interaction between tPA and NMDA receptor. I have thus shown that ischemic lesions decrease as a function of age, due to reduced levels of tPA. Moreover, I have identified DBP (D-site albumin Binding Protein), as being the transcription factor responsible for the control of tPA levels as a function of age. I have also developed a new model of thrombo-embolic ischemia in mice, in which tPA-induced thrombolysis is beneficial, provided it is performed soon enough. In this model, I have demonstrated by using a strategy of active immunization the in vivo occurrence of the cleavage of the NMDA receptor NR1 subunit by tPA. Finally, I have produced an antibody able to prevent the interaction between tPA and the NMDA receptor subunit, of which a single injection confers long lasting brain protection and neurological recovery and can also increase the therapeutic window of thrombolysis. This strategy could thus significantly increase the proportion of treatable ischemic stroke patients