The mismatching of alveolar ventilation and perfusion (VA/Q) is the major determinant of impaired gas exchange. The gold standard for analyzing VA/Q distribution is the multiple inert gas elimination technique (MIGET), conventionally based on gas chromatography (GC), and, although simple in principle, a technically demanding procedure limiting its use. A new technique based on micropore membrane inlet mass spectrometry (MMIMS) combined MIGET with mass spectrometry, simplifying the sample handling process, and potentially providing VA/Q distributions for a general clinical approach. The kinetics of volatile anesthetics are well known in patients with healthy lungs. The uptake and distribution of inhaled anesthetics have usually been modeled by physiologic models. However, these models have limitations, and they do not consider ventilation/perfusion matching. Respiratory diseases account for a large part of morbidity and mortality and are associated with pulmonary VA/Q mismatch that may affect uptake and elimination of volatile anesthetics. The objectives of the studies were firstly to investigate assessment of VA/Q mismatch by MMIMS and secondly to investigate the effects of asthma-like VA/Q mismatch on the kinetics of volatile anesthetics in an experimental porcine model. Anesthetized and mechanically ventilated piglets were studied. In study I, a direct comparison of MIGET by MMIMS with the conventional MIGET by GC in three animal models that covered a wide range of VA/Q distributions was preformed. The two methods agreed well, and parameters derived from both methods showed good agreement with externally measured references. In studies II–IV, a stable method of inducing and maintaining asthma-like VA/Q mismatch with methacholine (MCh) administration was established, and the effect of VA/Q mismatch on the pharmacokinetics of desflurane and isoflurane was investigated. The present model of bronchoconstriction demonstrates a delay in volatile anesthetic uptake and elimination, related to the heterogeneity of MCh-inhalation induced ventilation. The difference in solubility of volatile anesthetics has a significant influence on their uptake and elimination under VA/Q mismatch. The higher blood soluble isoflurane is affected to a lesser degree than the fairly insoluble desflurane.

La compréhension des mécanismes précis conduisant à la formation d'une plaque d'athérosclérose et à sa déstabilisation provoquant infarctus du myocarde ou accident vasculaire cérébral est fondamentale pour l'amélioration de la prévention, du dépistage et du traitement. Notre travail de thèse a porté sur l'analyse et le développement des modalités d'imagerie de la plaque d'athérosclérose afin d'améliorer notre capacité à suivre in vivo l'évolution de cette maladie. Nous proposons, après une introduction sur l'athérosclérose et la notion de plaque vulnérable, une revue des modèles animaux d'athérosclérose et l'intérêt du suivi en imagerie par résonance magnétique. Différentes modalités permettent le suivi en imagerie de la plaque d'athérosclérose. En pratique clinique, l'Imagerie par Résonance Magnétique (IRM) haute résolution est reconnue comme étant une modalité fiable et reproductible de caractérisation morphologique de la plaque carotidienne. Nous démontrons que l'injection de Gadolinium utilisée classiquement pour visualiser la lumière des vaisseaux permet également de préciser la caractérisation de la plaque carotidienne en donnant des informations sur la néovascularisation, la matrice extracellulaire et son statut inflammatoire. La caractérisation fonctionnelle de la plaque peut également être appréciée par l'injection de particules de fer en IRM ou par l'injection de radio-traceur comme le 18Ffluorodesoxyglucose en Tomographie par Emission de Positron (TEP). Le couplage de l'IRM et de la TEP de développement récent nous a permis de montrer une plus grande sensibilité de signal en TEP qu'en IRM avec particules de fer pour détecter les changements inflammatoires observés dans des plaques d'athérome d'aorte de lapin. Les applications actuelles de l'imagerie de l'athérosclérose sont nombreuses permettant de juger de l'efficacité thérapeutique d'un médicament ou d'identifier les patients à plus haut risque d'évènement cardiovasculaire / A better understanding of mechanisms leading to the formation of an atherosclerotic plaque and its destabilization causing myocardial infarction or stroke is fundamental to improve prevention, detection and treatment. Our work focused on development of imaging modalities of atherosclerotic plaque in order to improve our ability to monitor in vivo the progression of this disease. We propose, after an introduction to the concept of atherosclerosis and vulnerable plaque, a review of existing animal models of atherosclerosis and the interest of monitoring with magnetic resonance imaging. Different methods allow imaging of atherosclerotic plaque. In clinical practice, Magnetic Resonance Imaging (MRI) is recognized as a reliable and reproducible tool for morphological characterization of carotid plaque. We demonstrate that Gadolinium conventionally used to visualize the lumen of the vessels also allows a characterization of carotid plaque with information on neovascularization, extracellular matrix and inflammatory status. The functional characterization of the atherosclerotic plaque can be assessed with iron particles in MRI or with radiotracer such as 18Ffluorodeoxyglucose in positron emission tomography (PET). Combined PET-MR systems allowed us to show a better sensitivity of PET signal than USPIO-MR signal to detect early inflammatory changes in atherosclerotic plaque of rabbit aorta. We also provide some current clinical applications of atherosclerosis imaging (drug efficacy or identification of high risk patients)

INTRODUÇÃO: O modelo eslastase-induzido de aneurismas tem se destacado, nos últimos anos, porque simula as características geométricas dos aneurismas intracranianos humanos. A elastase destrói as fibras elásticas e dilata as artérias. A papaína é uma enzima que ainda não foi usada com esta finalidade. Os objetivos deste estudo foram determinar se a papaína produz aneurismas saculares em coelhos, e comparar suas características macroscópicas e histológicas com as dos aneurismas elastase-induzidos. MÉTODO: Dezoito coelhos brancos Nova Zelândia (1,9-3,2 kg) foram divididos em 3 grupos: I- elastase (n=7), II- papaína (n=8) e III- cirurgia controle (n=3). Os animais foram submetidos à exposição cirúrgica do pescoço, sendo que a artéria carótida comum direita foi usada como teste e a artéria carótida comum esquerda como controle. No 21° dia após a cirurgia, os animais foram sacrificados para retirada das artérias, tomada de suas medidas e análise histológica. Considerou-se formação de aneurisma quando a artéria teste dilatou em relação ao seu controle. RESULTADOS: Não houve aneurismas no grupo cirúrgico controle. Houve formação de aneurismas nos grupos elastase (71,4%) e papaína (100%). A diferença do diâmetro das artérias testes e seus respectivos controles não foi significativa (p= 0,15) entre os grupos elastase (média= 1,2 ± 0,4mm) e papaína (média= 2,1 ± 0,4mm), embora houvesse tendência deste último à maior dilatação . A histologia demonstrou que a papaína produziu maior tendência à lesão endotelial, à trombose (p = 0,01) e à inflamação parietal do que a elastase. A análise da fibrose intimal foi prejudicada em 50% dos casos do grupo papaína devido à trombose acentuada. Não houve diferença significativa entre os grupos quanto ao espessamento parietal (p=0,81) e ao grau de destruição das fibras elásticas (p= 0,009). CONCLUSÕES: A papaína produz aneurismas com tamanhos semelhantes aos da elastase, contudo a papaína provoca maior lesão endotelial, maior trombose e maior inflamação do que a elastase / INTRODUCTION: The elastase-induced model of experimental saccular aneurysms has been relevant in the last years because it mimics the size and geometric features of human intracranial aneurysms. Elastase destroys the arterys elastic fibers and produces arterial enlargement. Papain enzyme is also elastolytic but it had not been tested on saccular aneurysms creation yet. The purpose of this study was determine if papain produces saccular aneurysms in rabbits and to compare its gross and microscopic features are with the elastaseinduced aneurysms. METHODS: Eighteen New Zealand white rabbits (1.9 kg 3,2 kg) were separated in 3 groups: 1) sham (n=3) 0,9% saline solution; papain (n=8) 17-40 U; elastase (n=7) 6-8 U. The animals underwent surgical exposure of the neck; the right common carotid artery (RCCA) was used as the test and the left common carotid artery (LCCA) as the control. On the 21st day after surgery, animals were sacrificed for removal of the arteries, measurements and histological analysis. We determine formation of aneurysm to occur when the test artery dilated compared to the control. RESULTS: The sham group didnt develop aneurysms. There was aneurysm formation in the elastase (71,4%) and papain (100%) groups. The difference of the diameter of the tests and their respective controls is not significant (p=0,15) between elastase (average = 1,2 ± 0,4 mm) and papain (average = 2,1 ± 0,4mm) groups although there was tendency of this last one to produce larger aneurysms. the and to thrombosis (p = 0,01) and to parietal inflammation than the elastase. The analysis of the intimal fibrosis was not possible in 50% of papain cases due to pronounced thrombosis. There was no significant difference between the groups regarding the parietal thickening (p = 0,81) and the degree of destruction of the elastic fibers (0,009). CONCLUSION: Papain creates saccular aneurysms with similar dimensions to elastase-induced aneurysms. The microscopic results indicated papain destroys more endothelial cells, produces more thrombosis and more inflammatory process than elastase

Objetivo: Avaliar eletrofisiologicamente a função da retina do camundongo modelo de Alzheimer (3xTg-AD) comparando com seu controle (b6;129-PS1) em um estudo longitudinal com seis idades (2, 4, 6, 8, 10 e 12 meses). Métodos: Eletrorretinogramas (ERGs) foram registrados em 44 camundongos 3xTg-AD e em 23 controles, após administrada anestesia. Para o registro foi colocado um eletrodo de lente de contato sobre a córnea, um eletrodo de referência na cabeça e um terra na cauda. Em sessão de 30-40min de duração foram expostos ao seguinte protocolo de estimulação: 1) Adaptação ao escuro seguida de flashes nas intensidades: 0,003; 0,03; 0,3; 3 e 30 cd.s/m2; 2) Estimulação periódica (30 cd.s/m2) nas freqüências de 12, 18, e 30 Hz, sob luz de fundo (30 cd/m2). Resultados: Os ERGs mostraram dois tipos de respostas escotópicas tanto no grupo dos camundongos controles (b6;129- PS1) quanto nos modelos de Alzheimer. 13% dos camundongos controles e 72% dos modelos de AD apresentaram ERGs com potenciais oscilatórios presentes e tempo implícito da onda-b dentro da faixa esperada (45,31 ± 6,74 ms), enquanto no restante dos grupos, o ERG apresentou latência da onda-b muito aumentada (111,73 ± 22,56 ms) e potenciais oscilatórios ausentes. Devido a estes resultados, os grupos controle e experimental foram subdivididos em: b6;129 com OP, b6;129 sem OP; 3xTg-AD com OP e 3xTg-AD sem OP. Também foi incluído um grupo controle adicional constituído por 9 camundongos C57/B6. Comparando os cinco grupos, nenhuma diferença foi encontrada em relação à amplitude e à latência da onda-a. A amplitude da onda-b também foi semelhante para todos, ao contrário da latência para atingir o pico da onda-b dos grupos b6;129 sem OP e 3xTg-AD sem OP, que se apresentou duas vezes maior do que nos grupos com OP. As amplitudes dos cinco potenciais oscilatórios foram medidas individualmente e não mostraram diferenças entre os controles e os 3xTg-AD. Para o estímulo periódico, a amplitude do 1º harmônico dos grupos com OP mostrou clara diferença entre os grupos controle e o 3xTg-AD, tanto em 12 Hz como em 18 Hz. Os resultados dos dois grupos controle b6;129 e C57/B6 mantiveram-se muito próximos. Os grupos sem OP mantiveram-se sempre próximos a 10 V para as três freqüências de estimulação e mostraram atraso na diferença de fase média do 1º harmônico em 18 e 30 Hz, indicando maior lentidão de resposta, quando comparados aos primeiros. Conclusão: O camundongo 3xTg-AD e seu controle (b6;129) apresentam uma variante lenta e sem OPs do ERG escotópico em parte da população. Células bipolares, amácrinas e ganglionares podem estar alteradas nesses subgrupos (b6;129 sem OP e 3xTg-AD sem OP). Os grupos controle e 3xTg-AD com OPs diferiram quanto à amplitude de resposta à estimulação intermitente, diferença essa que implica em menor capacidade de processamento temporal para o modelo de AD. Sugerimos que as células bipolares de cones podem estar alteradas nos modelos de AD devido às amplitudes mais baixas dos 1os harmônicos desse grupo / Objective: To evaluate electrophysiologically the function of the retina of the Alzheimer model mouse (3xTg-AD) comparing it with its control (b6;129-PS1) in a longitudinal study at six ages (2, 4, 6, 8, 10 e 12 months) Methods: Electroretinograms (ERGs) were recorded in 44 anesthetized mice 3xTg-AD and in 23 controls, with a contact lens electrode placed on the cornea, a reference electrode on the head and a ground on the tail. During a 30-40min duration session the mice were exposed to the following stimulation protocol: 1) Scotopic response Dark adaptation followed by flashes at the following intensities: 0,003; 0,03; 0,3; 3 e 30 cd.s/m2; 2) Periodic stimulation (30 cd.s/m2) at the temporal frequencies of 12, 18, e 30 Hz, under background light (30 cd/m2). Results: The ERGs showed two types of scotopic responses, which ocurred in both the control mice (b6;129- PS1) and the Alzheimer´s models (3xTg-AD). 13% of the controls and 72% of the Alzheimer´s models mice presented ERGs with oscillatory potentials (OPs) and b-wave implicit times within the expected range (45,31 ± 6,74 ms), while for the other groups the ERG presented a very delayed b-wave latency (111,73 ± 22,56 ms) and absence of OPs. Given these results, the control and experimental groups were subdivided into: b6;129 with OPs, b6;129 without OPs; 3xTg-AD with OPs e 3xTg-AD without OPs. An additional control group with 9 mice C57/B6 was included. Comparing the five groups, no difference was found in a-wave amplitude and latency. The b-wave amplitude also did not differ among the groups, but the latency of the b-wave for the groups b6;129 without OPs and 3xTg-AD without OPs, was twice as long as in the groups with OPs. The amplitudes of the five OPs, measured individually, did not show differences between controls and 3xTg-AD groups. For the periodic stimulation the amplitude of the first harmonic of the Fourier transform of the groups with OPs showed a clear difference between the control and the 3xTg-AD groups, both for the 12 Hz and for the 18 Hz stimuli. The results of the two control groups (b6;129 and C57/B6) were very close. The groups without OPs had responses always close to 10 V for the three frequencies of stimulation and showed phase delay for the first harmonic, indicating response slowing, compared to the other groups. Conclusions: We found that a sub-group of both triple transgenic (3xTg-AD) and control mice (b6;129) manifest strikingly slow scotopic ERGs that lack OPs. We hypothesize that these response feature may reflect alterations in bipolar, amacrine and ganglion cells. The sub-group of triple transgenic and control mice that exhibited OPs differed in their response to flicker. Alzheimer model had significantly lower flicker-response amplitudes than the controls, suggesting impaired retinal temporal processing. We propose that the flicker results are consistent with alteration in cone bipolar cells in the Alzheimer model mice

Resultados recentes mostraram efeito ansiolítico do extrato hidroalcoólico bruto de Erythrina mulungu (EM) em ratos submetidos aos modelos do labirinto em T elevado (LTE) e da transição claro-escuro (TCE). O objetivo do presente trabalho foi realizar um estudo fitoquímico biomonitorado com a intenção de se verificar a participação dos alcalóides eritrínicos na atividade ansiolítica do extrato. Foi observado que o extrato bruto de EM apresentou efeito ansiolítico nas medidas de esquiva inibitória (100, 200 e 400 mg/Kg) e fuga (400 mg/Kg) dos braços abertos do LTE e na medida de tempo gasto pelos animais no compartimento iluminado (100 e 200 mg/Kg) no TCE. A partir do extrato hidroalcoólico de EM foi isolado um novo alcalóide eritrínico, a 11-hidroxi-eritravina, e dois já registrados na literatura, a eritrartina e a eritravina. Os testes com o LTE mostraram que a eritrartina, a eritravina (3 e 10 mg/Kg) e a 11-hidroxi-eritravina (10 mg/Kg), apresentaram efeito ansiolítico na medida de esquiva inibitória dos braços abertos. No TCE foi observado efeito ansiolítico com a eritravina (3 e 10 mg/Kg) e com a 11-hidroxi-eritravina (10 mg/Kg) na medida de tempo gasto pelos animais no compartimento iluminado. No TCE, a 11-hidroxi-eritravina (3 mg/Kg) também apresentou efeito ansiolítico aumentando o número de transição entre os dois compartimentos do modelo. Os efeitos ansiolíticos provocados pelos três alcalóides foram independentes de qualquer alteração locomotora, pois nenhum dos compostos estudados e, tampouco, o extrato bruto alterou o comportamento avaliado na arena. Desta forma, os resultados do presente trabalho mostraram que os alcalóides eritrartina, eritravina e 11-hidroxi-eritravina são responsáveis pelo efeito ansiolítico observado com o extrato bruto, o que explica a ampla utilização popular das plantas do gênero Erythrina como \"calmante\". / One new erythrinian alkaloid derivative 11-OH-erythravine (3) and the known erythravine (2), and erythrartine (1) from Erythrina mulungu were isolated, and their structures were determined by spectral analysis. The relative stereochemistry of the new alkaloid was determined mainly by 1H NMR experiments, including NOESY spectrometry. Furthermore, the anxiolytic properties of the hydroalcoholic crude extract (CE) and of the alkaloids were evaluated using the elevated plus-maze test (EPM), the elevated T-maze test (ETM) and the light-dark transition model (LDTM) for mice. The CE showed anxiolytic-like effects in the ETM, i.e., the doses 100, 200, and 400 mg/kg (po) of CE impaired the inhibitory avoidance of the open arms of the maze. In the LDTM, CE (100 and 200 mg/kg) increased the time spent by the animals in the illuminated compartment, an anxiolytic-like effect. Since CE did not alter the anxiety related responses in mice exposed to the EPM, we did not use this model to test the anxiolytic-like effects of the erythrinian alkaloids. Interestingly, the compounds 1, 2, and 3 also attenuated anxiety in the ETM, an effect that was confirmed in the LDTM with the alkaloids 2 and 3. Taken in account, these results strongly suggest that these erythrinian alkaloids are the compounds responsible for the anxiolytic properties attributed to the crude extract and seem to supports the folk medicinal use as a natural anxiolytic.

Resultados recentes mostraram efeito ansiolítico do extrato hidroalcoólico bruto de Erythrina mulungu (EM) em ratos submetidos aos modelos do labirinto em T elevado (LTE) e da transição claro-escuro (TCE). O objetivo do presente trabalho foi realizar um estudo fitoquímico biomonitorado com a intenção de se verificar a participação dos alcalóides eritrínicos na atividade ansiolítica do extrato. Foi observado que o extrato bruto de EM apresentou efeito ansiolítico nas medidas de esquiva inibitória (100, 200 e 400 mg/Kg) e fuga (400 mg/Kg) dos braços abertos do LTE e na medida de tempo gasto pelos animais no compartimento iluminado (100 e 200 mg/Kg) no TCE. A partir do extrato hidroalcoólico de EM foi isolado um novo alcalóide eritrínico, a 11-hidroxi-eritravina, e dois já registrados na literatura, a eritrartina e a eritravina. Os testes com o LTE mostraram que a eritrartina, a eritravina (3 e 10 mg/Kg) e a 11-hidroxi-eritravina (10 mg/Kg), apresentaram efeito ansiolítico na medida de esquiva inibitória dos braços abertos. No TCE foi observado efeito ansiolítico com a eritravina (3 e 10 mg/Kg) e com a 11-hidroxi-eritravina (10 mg/Kg) na medida de tempo gasto pelos animais no compartimento iluminado. No TCE, a 11-hidroxi-eritravina (3 mg/Kg) também apresentou efeito ansiolítico aumentando o número de transição entre os dois compartimentos do modelo. Os efeitos ansiolíticos provocados pelos três alcalóides foram independentes de qualquer alteração locomotora, pois nenhum dos compostos estudados e, tampouco, o extrato bruto alterou o comportamento avaliado na arena. Desta forma, os resultados do presente trabalho mostraram que os alcalóides eritrartina, eritravina e 11-hidroxi-eritravina são responsáveis pelo efeito ansiolítico observado com o extrato bruto, o que explica a ampla utilização popular das plantas do gênero Erythrina como \"calmante\". / One new erythrinian alkaloid derivative 11-OH-erythravine (3) and the known erythravine (2), and erythrartine (1) from Erythrina mulungu were isolated, and their structures were determined by spectral analysis. The relative stereochemistry of the new alkaloid was determined mainly by 1H NMR experiments, including NOESY spectrometry. Furthermore, the anxiolytic properties of the hydroalcoholic crude extract (CE) and of the alkaloids were evaluated using the elevated plus-maze test (EPM), the elevated T-maze test (ETM) and the light-dark transition model (LDTM) for mice. The CE showed anxiolytic-like effects in the ETM, i.e., the doses 100, 200, and 400 mg/kg (po) of CE impaired the inhibitory avoidance of the open arms of the maze. In the LDTM, CE (100 and 200 mg/kg) increased the time spent by the animals in the illuminated compartment, an anxiolytic-like effect. Since CE did not alter the anxiety related responses in mice exposed to the EPM, we did not use this model to test the anxiolytic-like effects of the erythrinian alkaloids. Interestingly, the compounds 1, 2, and 3 also attenuated anxiety in the ETM, an effect that was confirmed in the LDTM with the alkaloids 2 and 3. Taken in account, these results strongly suggest that these erythrinian alkaloids are the compounds responsible for the anxiolytic properties attributed to the crude extract and seem to supports the folk medicinal use as a natural anxiolytic.

Introdução: A perda da função esfincteriana anal e a colostomia definitiva são condições socialmente incapacitantes, com grande impacto social e econômico. Não há tratamento satisfatório para estas duas condições e o transplante anorretal surge como uma alternativa para a perda da função ou do esfíncter anal propriamente dito. Objetivo: apresentar modelo experimental de autotransplante anorretal em ratos, avaliar os seus resultados clínicos e funcionais Método: 55 ratos Wistar foram divididos aleatoriamente em quatro grupos de estudo: I autotransplante anorretal ortotópico (n=13); II - autotransplante heterotópico (n=14); III - SHAM - ratos submetidos a laparotomia e incisão perianal (n=13); IV - Controle - ratos não operados (n=15). Os animais foram avaliados com parâmetros clínicos e manométricos. Os parâmetros clínicos de avaliação foram: peso, comportamento, aspecto das fezes, aspecto anal e aspecto abdominal. Os dados de avaliação clínica foram protocolados no dia do procedimento (D0), segundo dia pós-operatório (D2), sétimo (D7) e décimo quarto dias (D14). Exceto peso, todos os demais foram avaliados por escores préestabelecidos. A manometria foi usada para determinação das pressões Médias e Máximas, e realizadas no pré-operatório imediato (D0 pré), logo após o procedimento (D0 pós), no D7 e no D14. O grupo II teve avaliação manométrica apenas no D0 pré e D14. As manometrias de ratos normais foram analisadas separadamente, para determinação do perfil funcional anorretal normal. Foram também comparados os grupos I, III e IV em todos os momentos, e os quatro grupos nos momentos D0 pré e D14. Os animais foram sacrificados após 14 dias e o segmento anorretal foi analisado histologicamente. Resultados: ocorreram nove óbitos pós operatórios, 5 no grupo II e 4 no grupo I. Todos os parâmetro clínicos mostraram tendência de piora no pós-operatório precoce nos grupos transplantados. Porém, houve equiparação entre os grupos ao final do estudo. A anatomia patológica demonstrou histologia análoga entre os grupos transplantados e os controles após 14 dias. As manometrias em ratos normais tiveram média de pressões Médias de 33,7 ± 12,6 cmH2O, e de pressões Máximas de 61,2 ± 19 cmH2O. Na comparação entre os grupos I, III e IV, no D0 pós existiu queda significativa das pressões nos grupos I e III, diferentes do IV (p<0,05). No D7 observou-se recuperação de atividade manométrica do grupo I, porém estatisticamente inferior ao grupo Controle. No D14 os grupos foram estatisticamente semelhantes. Na comparação dos quatro grupos houve queda pressórica significativa no grupo II, que diferiu dos grupos controle (p<0,05). O grupo I não diferiu dos controles nem do grupo II no D14. Conclusões: 1. O modelo experimental proposto de autotransplante anorretal ortotópico e heterotópico em ratos é factível. 2. A evolução clínica e funcional do transplante anorretal demonstrou perda da função imediatamente após o transplante, com recuperação ao longo de 14 dias. 3. O transplante heterotópico teve resultados precoces piores que o ortotópico / Introduction: The loss of anal sphincter function and permanent colostomy are disabling conditions with great social and economic impact. There is no satisfactory treatment for either of these conditions and anorectal transplantation offers an alternative to the loss of function or the loss of the anal sphincter itself. Aim: To present an experimental model of anorectal autotransplantation in rats to assess the clinical and functional results. Method: 55 Wistar rats were divided randomly into four study groups: I - orthotopic anorectal autotransplantation (n=13), II - Heterotopic autotransplantation (n=14), III SHAM - Laparotomy and perianal incision (n=13), IV - Control non-operated rats (n=15). The animals were evaluated with clinical and manometric parameters. The clinical parameters of evaluation were: weight, behavior, appearance of stools, appearance of the anus and abdominal distension. Clinical trial data was collected on the day of the procedure (D0) and the second (D2), seventh (D7) and fourteenth (D14) post-operative days. All parameters, except for weight, were evaluated by pre-established scores. Manometry was used to determine the Mean and Maximum pressures, and performed immediately before (D0 pre), immediately after the procedure (D0 post), and on D7 and D14. Group II had manometric evaluation only on D0 pre and D14. The manometries of the non-operated rats were analyzed separately to determine a normal anorectal functional profile. Groups I, III and IV were also compared at all evaluation times and all four groups on D0 pre and D14. The animals were sacrificed after 14 days and the anorectal segment was examined histologically. Results: There were nine post-operative deaths, 5 in group II and 4 in group I. All clinical parameters showed a worsening trend in the early postoperative evaluation in the transplant groups. However, at the end of the study there were no significant differences between the groups. The histology of the transplanted groups and the control groups was similar after 14 days. The manometries in normal rats had an average of 33.7 ±12.6 cmH2O Mean pressure, and average Maximum pressure of 61.2 ±19 cmH2O. In the analysis of groups I, III and IV, a significant drop of pressure in groups I and III occurred at D0 post compared to group IV (p<0.05). Functional recovery of Group I was noted in D7, but statistically lower than the Control group. The three groups were statistically similar at D14. In the comparison of the four groups there was a significant pressure fall in Group II, which differed from the control groups (p<0.05). Group I did not differ from the controls or Group II in D14. Conclusions: 1.The proposed experimental orthotopic and heterotopic anorectal autotransplantation model in rats is feasible. 2. The clinical and functional outcomes demonstrated loss of function immediately after transplantation, with recovery over the course of 14 days. 3. The Heterotopic transplantation had worse early results than the orthotopic method

吸煙是引起骨質疏鬆症的因素之一。臨床研究清楚表明吸煙者的骨密度降低，但其他干擾因素可能掩蓋了吸煙對骨頭的不良效果。使用動物模型用以研究吸煙和骨質疏鬆症之間是否有直接的因果關係與它潛在的機制是有必要的。為此，我們使用年輕和雌激素耗盡的小鼠作被動吸煙模型以及小鼠成骨細胞和破骨細胞株來作研究。 / 年輕的Balb/c小鼠暴露於2%或4% (v/v)的香煙煙霧中，代表中度和重度吸煙的人。骨代謝生物標誌物明顯增加，4%吸煙組在14週後股骨的微觀結構4%顯著下降，這相等於人類吸煙12年。此外，雌性Balb/c小鼠進行4%吸煙和/或卵巢切除術(OVX)。吸煙+OVX組增加血清中骨轉換指標水平。4%吸煙組的股骨生長板較薄。μ-CT數據進一步表明，相對骨體積(BV / TV)和結構模型指數(SMI)在吸煙組有顯著影響，而且在吸煙+ OVX組上有更大程度的影響。 / 在細胞研究中使用氯仿(CSE)和乙醇的香煙提取物(ESE)。CSE抑制小鼠細胞株RAW 264.7形成破骨細胞，並刺激小鼠成骨細胞株的分化和功能。這個與體內研究矛盾的結果暗示直接從煙霧中提取的化學成分並不是引起骨質疏鬆的元兇。影響骨代謝的很可能是其他從煙霧中生成的活性代謝物和一些吸煙引起的內源性激素物質。在吸煙引起的骨質流失中，這些代謝物或內源性物質可能是非常重要的。 / 有見及此，4%吸煙小鼠的血清用以研究其對成骨細胞和破骨細胞活動的影響。吸煙小鼠血清顯著降低在成骨細胞中鹼性磷酸酶(ALP)活性和鈣沉積，一些成骨細胞標記基因和蛋白表達均下降，而且 Wnt/β-catenin信號通路下調。此外，吸煙小鼠血清顯著增加形成破骨細胞的數量，組織蛋白酶K的基因和蛋白表達增加，在NF-κB和p-38 MAPK信號傳導途徑的信號分子表達增加。 / 總而言之，大量吸煙可能影響年輕小鼠和雌激素耗竭小鼠的骨代謝和微結構，通過類似的行動機制，人類也可能有同樣的骨疾病風險。這項研究揭示了吸煙導致的骨質疏鬆症在青少年和絶經後婦女的發病機制。這也給我們線索如何預防和治療與吸煙有關的骨骼疾病。這項研究還傳達了一個明確的信息：在年輕時應開始應控制吸煙。 / Cigarette smoking is one of the risk factors for osteoporosis. Clinical studies clearly showed that smokers have lower bone mineral density, but other confounding factors could mask the adverse actions of smoking on bones. Animal models are warranted to study the direct causal relationship between cigarette smoking and osteoporosis, and also the underlying mechanisms. In this regard, we used a mouse passive smoking model in both young and estrogen depleted mice, and the mouse osteoblast and osteoclast cell lines. / Young Balb/c mice were exposed to 2 or 4% (v/v) of cigarette smoke, similar to moderate or heavy smoking respectively in humans. Biomarkers for bone turnover were increased and bone microstructure of femur was significantly deteriorated after 4% smoking for 14 weeks which is similar to cigarette smoking for 12 years in humans. Furthermore, the effects of heavy smoking on ovariectomized mice were also investigated. Female Balb/c mice were subjected to 4% cigarette smoking and/or ovariectomy (OVX). Cigarette smoking together with OVX further increased the levels of bone turnover markers in serum. Femur growth plate was thinner in the 4% smoking group when compared to those in the SHAM- and OVX-operated groups. Micro-CT data further indicated that the relative bone volume (BV/TV) and structural model index (SMI) were significantly affected in the smoking groups, and to a greater extent in the 4% smoking + OVX group. / Chloroform (CSE) and ethanol smoke extracts (ESE) were used in cell studies. CSE suppressed the formation of osteoclasts, and stimulated the differentiation and function of mouse osteoblasts. These findings are contradictory to those found in in vivo study implying that chemical components directly extracted from cigarette smoke are not the culprits in causing bone disorder in animals. It is likely that other active metabolites from cigarette smoke and some endogenous hormonal substances released by cigarette smoking could affect bone metabolism. These active metabolites together with the endogenous bone hormones are perhaps crucial in smoking-induced bone loss in the body. / In view of this hypothesis, sera from 4% smoking mice were used to investigate their effects on osteoblast and osteoclast activities. It was found that the alkaline phosphatase (ALP) activity and calcium deposition in osteoblast were reduced significantly by the sera from smoking mice. Gene and protein expressions of some osteoblast markers were also decreased. The downregulation of Wnt/β-catenin signaling pathway was observed after the treatment with the serum obtained from the 4% smoking group. Moreover, the number of osteoclasts being formed was increased significantly by the smoking mouse serum. Cathepsin K gene and protein expressions were also induced. The increased expressions of the signaling molecules including NF-κB and p-38 MAPK were also observed. / In conclusion, heavy cigarette smoking could deteriorate bone metabolism and microstructures in young female and also estrogen depleted mice. The same kind of risk in bone disease may also apply to humans through similar mechanisms of action. This study sheds light in understanding the pathogenesis of smoking-induced bone disorders in teenagers and also postmenopausal women. It also gives us clues how to prevent and treat smoking related bone diseases. This study also conveys a clear message that cigarette smoking control should be started in young ages. / Detailed summary in vernacular field only. / Detailed summary in vernacular field only. / Detailed summary in vernacular field only. / Detailed summary in vernacular field only. / Detailed summary in vernacular field only. / Chan, Lok Yi Ruby. / Thesis (Ph.D.) Chinese University of Hong Kong, 2014. / Includes bibliographical references (leaves 182-207). / Abstracts also in Chinese. / Chan, Lok Yi Ruby.

Les accidents vasculaires cérébraux sont la première cause mondiale d'handicap et l'athérosclérose en est le principal facteur. Cette pathologie, liée à une mauvaise prise en charge du cholestérol pourra avoir des conséquences plus pernicieuses comme la fragilisation des unités cérébrovasculaires qui, combinée à une inflammation systémique et locale, peut entraîner d'importantes répercussions cérébrales.Pour être au plus proche de l'humain nous avons utilisé des modèles animaux murins et primate non-humain (PNH) âgés sous régime gras. Une approche translationnelle avec suivi longitudinal de biomarqueurs sanguins et d'imagerie combinée à la caractérisation tissulaire de l'inflammation a été effectuée pour tenter d'élucider les spécificités de la réponse inflammatoire dans la paroi vasculaire des grosses artères et le tissu cérébral.Nous avons montré que chez des souris ApoE-/- âgées l'exercice physique peut contrecarrer les effets délétères d'un régime gras lorsque l'apport calorique est contrôlé mais plus lorsqu'il ne l'est pas. La dégradation de la barrière hémato-encéphalique pourrait expliquer l'inflammation observée in vivo et confirmée par l'analyse tissulaire. L'étude des PNH a montré l'intérêt d'associer imagerie multimodale et dosages sanguins dans la stratification du risque cardiovasculaire ainsi que l'importance d'associer des marqueurs métaboliques, inflammatoires et anti-inflammatoires.Nous avons montré l'intérêt de contrôler les apports caloriques pour bénéficier des effets protecteurs de l'exercice sur l'athérosclérose et l'importance d'avoir une vue globale du patient pour une stratification individuelle précise du risque cardio et cérébrovasculaire / Stroke is the leading cause of disabilities worldwide and is mainly caused by atherosclerosis. But this is not the only risk for patients. Indeed, as this pathology is due to a lack of circulating cholesterol management and could lead to more pernicious outcomes such as cerebrovascular units disorganization whom, when combined with systemic and local inflammation can result in serious brain aftermath.Aged murine and non-human primate (NHP) animal models under high cholesterol diet were used to be closest to the human pathology. A translational approach with longitudinal follow-up of circulating and imaging biomarkers combined with a tissular characterization of inflammation was performed in order to elucidate the specificities of the inflammatory response in vascular wall of large vessels and brain tissue.We showed that in old ApoE-/- mice exercise can counterbalance deleterious effects of high fat diet when caloric intake is controlled but no longer when food is given ad libitum. The leakage of blood-brain barrier might explain the neuroinflammation observed in vivo and confirmed by tissular analysis. The study on NHP showed the interest of combine multimodal imaging with blood dosage for cardiovascular risk stratification and the importance of associating metabolic, inflammatory but also anti-inflammatory markers.We highlighted the importance of controlling calories intake in order to benefit of protective effects of exercise on atherosclerosis and the relevance of having an overview of the patient’s status for an accurate individual stratification of cardio and cerebrovascular risk

Quando as escoriações ocorrem é esperada uma cicatrização sem complicações, utilizando produtos para prevenção de infecções e que proporcionam ambiente que otimiza rápida cicatrização com cicatrizes mínimas. Tratamentos com antissépticos tópicos e produtos cicatrizantes são os mais utilizados. Vários relatos demonstram que o soro do látex (SLX) da seringueira Hevea brasiliensis tem propriedades cicatrizantes para úlceras cutâneas. O objetivo deste trabalho foi avaliar citotoxicidade e potencial proliferativo do SLX em ensaios in vitro e sua eficácia em escoriações cutâneas no dorso de ratos. A citotoxicidade do SLX (10%, 1% e 0,1%) foi avaliada em culturas de fibroblastos 3T3 pela determinação da viabilidade celular. A proliferação/migração celular com queratinócitos humanos foi avaliada pelo método scratch assay nas concentrações (1%, 0,1%, 0,01%, 0,001%, 0,0001% e 0,00001%). Para a atividade cicatrizante utilizou-se 72 ratos Wistar submetidos à escoriação no dorso por dermoabrasão, e 3 grupos foram estabelecidos: soro fisiológico (SF), antisséptico (AS-Merthiolate®) e soro do látex (SLX-Regederm®), aplicado diariamente por 10 dias. As lesões foram fotografadas e avaliadas por análise de imagem nos dias 2, 7 e 10 póslesão e calculados os índices de cicatrização das escoriações (ICE) para medir reepitelização. Nos dias 2, 7 e 10 foram sacrificados 8 animais de cada grupo e coletadas amostras para dosagem bioquímica da mieloperoxidase (MPO) e no 10º dia uma parte das biópsias foram separadas para análise histológica da epiderme e crosta. O SLX não apresentou toxicidade aos fibroblastos nas concentrações menores ou iguais à 1%, sendo inviável na concentração de 10%. Pelo scratch assay o SLX apresentou atividade proliferativa sobre queratinócitos humanos principalmente na concentração de 0,01% (89%). O SLX promoveu melhor reepitelização no modelo de escoriação, principalmente no 7º dia, diferente dos demais grupos que tinham atraso na reepitelização. Além disso, quanto às diferenças de espessura da crosta e da epiderme, o grupo SLX promoveu um aumento no número de camadas epidérmicas e menor espessura das crostas. Os grupos SLX e AS apresentaram aumento da enzima MPO no 2º dia, porém com redução significante a partir do 7º dia, diminuindo assim a inflamação e acelerando consequentemente a reepitelização. Os resultados evidenciaram que a aplicação do soro do látex é segura pela não toxicidade e comprova sua eficácia perante a outros produtos, pela propriedade proliferativa para queratinócitos e pela aceleração da cicatrização em modelos de exulcerações cutâneas. / When abrasions occur is expected to heal without complications, using products to prevent infections and provide environment that optimizes rapid healing with minimal scarring. Treatments with topical antiseptics and healing products are the most used. Several reports show that the latex serum (LXS) from Hevea brasiliensis has healing properties for skin ulcers. The objective was to evaluate the cytotoxic and proliferative potential of LXS in vitro assays and their efficacy to heal skin abrasions on the back of rats. LXS cytotoxicity (10, 1 and 0.1%) was evaluated in 3T3 fibroblast cultures by cell viability. The cell proliferation/migration in human keratinocytes was evaluated by the scratch assay method using 1%, 0.1%, 0.01%, 0.001%, 0.0001% and 0.00001% concentrations. For healing activity, 72 Wistar rats were underwent to dermoabrasion on the back and 3 groups were stablished: saline (S), antiseptic (AS-Merthiolate®) and rubber latex serum (LXS-Regederm®). The different products were applied daily for 10 days. The lesions were photographed and evaluated by image analysis on days 2, 7 and 10 post-injury and the abrasion healing rate (AHR) were calculated for evaluation of the re-epithelialization. On days 2, 7 and 10, 8 animals from each group were sacrificed, and samples were taken for biochemical determination of myeloperoxidase (MPO) After that, on day 10, a part of the biopsies was separated for histological analysis of the epidermis and crust. The LXS showed no toxicity to fibroblasts at concentrations less than or equal 1%, being impractical at a concentration of 10%. By the scratch assay, the LXS showed proliferative activity on human keratinocytes, particularly at a concentration of 0.01% (89%). The LXS produced better re-epithelialization in the excoriation model, especially on the 7th day, unlike other groups, in which the re-epithelialization was delayed. Furthermore, it showed significant differences regarding the amount of epidermis and crust. The Regederm group presented an increase in the number of epidermal layers and thinner thickness of their crusts. The LXS and AS groups showed an increase of MPO enzyme on the 2nd day, however, with a significant decrease from the 7th day, thus reducing inflammation and consequently accelerating re-epithelialization. The results showed that the application of the rubber latex serum is safe due its non-toxicity and prove its efficacy comparing to other products, because of its proliferative property for keratinocytes and acceleration of healing in cutaneous exulceration models.