Lacunar strokes account for approximately a quarter of all ischaemic strokes and traditionally are thought to result from occlusion of a small deep perforating arteriole in the brain. Lacunar infarcts can be up to 2cm in diameter and are found in deep brain structures such as the thalamus and internal capsule. Despite their prevalence and specific accompanying clinical syndromes, the cause of lacunar stroke and its associated vascular pathology remain unclear. Many hypotheses as to the cause exist, which fall broadly into two categories; firstly, a direct occlusion via emboli or thrombus usually from a cardiac or large artery source, microatheroma (intrinsic lenticulostriate occlusion) or macroatheroma (parent artery occlusion) all operating primarily via ischaemia. Secondly, there could be an indirect occlusion resulting from vasospasm, endothelial dysfunction or other forms of endovascular damage (e.g. inflammation). Therefore the question of whether the resulting lesions are truly “ischaemic” or actually arise secondary to an alternative process is still under debate. To clarify the chain of pathological events ultimately resulting in lacunar stroke, in this thesis I firstly undertook a systematic assessment of human lacunar stroke pathology literature to determine the information currently available and the quality of these studies (including terminology). The majority of these studies were performed in patients who had died long after their stroke making it difficult to determine the early changes, and there were few patients with a clinically verified lacunar syndrome. Therefore I adopted alternative approaches. In this thesis, I systematically looked for all potential experimental models of lacunar stroke and identified what appears at present to be the most pertinent - the spontaneous pathology of the stroke-prone spontaneously hypertensive rat (SHRSP). However, the cerebral pathology described in this model to date is biased towards end stage pathology, with little information concerning the microvasculature (as opposed to the brain parenchyma) and confounding by use of salt to exacerbate pathology. Therefore, the aim of the experimental work in this thesis was to assess pathological changes within the cerebral vasculature and brain parenchyma of the SHRSP across a variety of ages (particularly young pre-hypertensive animals) and to look at the effects of salt loading on both the SHRSP and its parent strain (the Wistar Kyoto rat - WKY). Three related studies (qualitative and quantitative histology, immunohistochemistry and a microarray study of gene expression confirmed by quantitative PCR), revealed that the presence of inflammation (via significant changes in gene expression in the acute phase response pathway and increased immunostaining of activated microglia and astrocytes) plus alterations in vascular tone regulation, (via genetic alteration of the nitric oxide signaling pathway probably secondary to abnormal oxidative state), impaired structural integrity of the blood brain barrier (histological evidence of endothelial dysfunction and significantly decreased Claudin-5 staining) and reduced plasma oncotic potential (reduced albumin gene expression) are all present in the native SHRSP at 5 weeks of age, i.e. well before the onset of hypertension and without exposure to high levels of salt. We also confirmed previous findings of vessel remodelling at older ages likely as a secondary response to hypertension (thickened arteriolar smooth muscle, increased smooth muscle actin immunostaining). Furthermore, we found not only that salt exacerbated the changes see in the SHRSP at 21 weeks, but also that the control animals (WKY) exposed to a high salt intake developed features of cerebral microvascular pathology independently of hypertension (e.g. white matter vacuolation and significant changes in myelin basic protein expression). In conclusion, via the assessment of the most pertinent experimental model of lacunar stroke currently available, this thesis has provided two very important pieces of evidence: firstly that cerebral small vessel disease is primarily caused by a non-ischaemic mechanism and that any thrombotic vessel lesions occur as secondary end stage pathology; secondly that these features are not simply the consequence of exposure to raised blood pressure but occur secondary to abnormal endothelial integrity, inflammation, abnormal oxidative pathways influencing regulation of vascular tone and low plasma oncotic pressure. Patients with an innate susceptibility to increased blood brain barrier permeability and/or chronic inflammation could therefore have a higher risk of developing small vessel disease pathology and ultimately lacunar stroke and other features of small vessel disease. Research, addressing whether lacunar stroke patients should be treated differently to those with atherothromboembolic stroke is urgently needed.

Einleitung Der Einsatz von Stammzellen als Grundlage neuer therapeutischer Strategien wird bereits seit über 25 Jahren intensiv erforscht. Stammzellen sind in der Lage, in verschiedene funktionale Zelltypen auszudifferenzieren und verfügen über ein enormes Proliferationspotential (NAM et al. 2015). Ausgehend von den Fähigkeiten von Stammzellen sehen Forscher und Kliniker erstmals eine realistische Möglichkeit, kurative Therapieoptionen für Erkrankungen zu entwickeln, die bisher als schwer behandelbar oder sogar unheilbar angesehen wurden. Davon könnten insbesondere Patienten chronisch-degenerativer neurologischer und zerebrovaskulärer Erkrankungen, einschließlich der großen Anzahl an Schlaganfallopfern, profitieren. Schlaganfälle repräsentieren eine der häufigsten Todesursachen in der westlichen Welt (LOPEZ et al. 2006). Ein Drittel der betroffenen Patienten verstirbt innerhalb eines Jahres, während etwa 40% von dauerhaften Behinderungen betroffen sind (MOZAFFARIAN et al. 2015). Trotz intensiver Forschung existieren neben der systemischen Thrombolyse, die auf einen engen Zeitraum von maximal 4,5 Stunden nach dem Akutereignis beschränkt ist, keine zugelassenen Therapieoptionen (HACKE et al. 2008, SAVER et al. 2009). Zelltherapeutische Strategien zur Behandlung des Schlaganfalls werden daher als besonders vielversprechend angesehen (ANDRES et al. 2011). Neben den bereits gesicherten Erkenntnissen zur stammzelltherapeutischen Sicherheit und Wirksamkeit aus Studien unter Einsatz gängiger Nagermodellen (BLISS et al. 2006, JOO et al. 2013) wird insbesondere die Überprüfung der Wirksamkeit an geeigneten Großtiermodellen gefordert, die die Situation des menschlichen Schlaganfallpatienten möglichst realistisch wiedergeben sollen (SAVITZ et al. 2011). Eine Voraussetzung für die erfolgreiche Testung eines zelltherapeutischen Ansatzes in einem Großtiermodell mit fokaler zerebraler Ischämie besteht darin, ein langfristiges Überleben xenogener Zelltransplantate durch ein geeignetes Immunsuppressionsprotokoll zu erreichen. Die Notwendigkeit einer Immunsuppression besteht darin, dass sowohl allo- als auch xenogene Transplantate eine Immunantwort beim Empfänger auslösen und somit zu einer Abstoßungsreaktion führen können (JANEWAY 2002). Die Anwendung von immunsuppressiven Medikamenten geht dabei aber häufig mit Nebenwirkungen einher. Insbesondere beim Schaf existiert jedoch nur eine limitierte Datenlage zu immunsuppressiven Protokollen und deren Nebenwirkungen. Ziele der Untersuchung Das Ziel der vorliegenden Studie bestand darin, eine xenogene Transplantation von fetalen humanen neuralen Progenitorzellen (fhNPZ) in einem gesunden Schafsmodell durchzuführen, um die Wirksamkeit in Hinblick auf das Transplantatüberleben und die Nebenwirkungen einer Immunsuppression mittels Cyclosporin A (CsA) zu untersuchen. Materialien und Methoden Hierfür wurden je 5 Schafe in zwei Gruppen über einen Zeitraum von 64 Tagen immunsupprimiert (iCsA: 3 mg CsA/kg 2x tägl. bis einschließlich Tag 36, danach 3 mg CsA/kg 1x tägl. jeden 3. Tag; kCsA: kontinuierlich 3 mg CsA/kg 2x tägl.), während eine Kontrollgruppe (Kon) von ebenfalls 5 Tieren keine Immunsuppression erhielt. Am Versuchstag 22 wurde den Schafen eisenmarkierte fhNPZ (Eisenkonzentration: 3,0 mM, ca. 200.000 Zellen pro Transplantationsposition) stereotaktisch in das gesunde Gehirn transplantiert. Aufgrund der Eisenmarkierung der Stammzellen konnten diese an den Versuchstagen 23, 36 und 64 mittels 3,0 MRT-Aufnahmen in vivo überwacht und anschließend ex vivo das Überleben der fhNPZ im Schafhirn 42 Tage nach Transplantation histologisch untersucht werden. Für die Untersuchungen zu Wirkspiegeln und Nebenwirkungen von CsA im Schaf wurden den Versuchstieren innerhalb des Versuchszeitraums regelmäßig Blutproben entnommen und am Versuchsende eine pathologische und histologische Untersuchung von Leber und Nieren durchgeführt. Ergebnisse Bei den durchgeführten Untersuchungen konnte festgestellt werden, dass die CsA-Wirkspiegel im Blut bei der kCsA (424,0 ± 135,0 ng/ml) signifikant höher waren im Vergleich zur iCsA (198,5 ± 155,9 ng/ml). Diese Unterschiede besaßen jedoch keinen Einfluss auf das Langzeitüberleben der transplantierten fhNPZ. In keiner der drei Versuchsgruppen konnten vitale Zellen 42 Tage nach der Transplantation aufgefunden werden. Die Untersuchung der Nebenwirkungen von CsA ergab, dass die Langzeitgabe von CsA Anzeichen für einen hämatologischen Einfluss zeigt. Ebenso konnte sowohl eine hepatotoxische, als auch eine nephrotoxische Wirkung von CsA beim Schaf nachgewiesen werden. Schlussfolgerungen Schlussfolgernd kann zusammengefasst werden, dass die Gabe von 3 mg CsA/kg 2x tägl. nicht suffizient einer Abstoßungsreaktion xenogener ins Schafhirn transplantierter fhNPZ entgegenwirkt. Für das Ziel einer suffizienten zelltherapeutischen Anwendung im Schaf nach einem Schlaganfall sind somit weitere Untersuchungen zu einer wirksamen Immunsuppression beim Schaf und zu einem verbesserten Transplantatüberleben notwendig. Desweiteren konnten klinische und pathologische Nebenwirkungen beim Schaf durch die Langzeitgabe des Immunsuppressivums CsA festgestellt werden.

English Abstract Glutamate carboxypeptidase II (GCPII) is a membrane metallopeptidase expressed in many human tissues, predominantly in prostate, brain and small intestine. In brain it cleaves the most abundant peptide neurotransmitter N-acetyl-L-aspartyl-α-L-glutamate into N-acetyl-L-aspartate and free L-glutamate. Thus, GCPII participates in glutamate excitotoxicity through the release of free glutamate into the synaptic cleft. Inhibition of this activity has been shown to be neuroprotective in rats. In the human jejunal brush border, GCPII cleaves off terminal glutamate moieties from poly-γ-glutamylated folates, which can be then transported across the intestinal mucosa. The function of GCPII in human prostate is unknown but it is overexpressed in prostate cancer. Therefore, GCPII is an important marker of prostate cancer and its progression.Moreover, it could become a perspective target for treatment of prostate cancer as well as neuronal disorders associated with glutamate excitotoxicity. For the development and testing of novel drugs and therapeutics it is necessary to have an appropriate animal model. Mouse (Mus musculus) is such a model and it is widely used by many experimentators. However, no detailed comparison of mouse and human GCPII orthologs regarding their enzymatic activity, inhibition...

Title: Structural changes in model of hypoxic-ischemic encephalopathy in rat Objectives: The aim of the research is to develop a model of hypoxic-ischemic encephalopathy in a rat that would represent perinatal injury in a human and then histologically differentiate the most commonly damaged cerebral structures. Methods: This is an experimental study. Five laboratory rats underwent hypoxic-ischemic conditions causing encephalopathy according to the Rice-Vannucci model. The control group representing the other five rats underwent hypoxia for 1.5 hours. Subsequently, the animals were returned to their mother. 48 hours later, cerebral perfusion, paraffinisation, slicing the brain into sections and followed by applying these sections onto slides. Sections to represent morphological changes and degeneration of neurons were stained with Hematoxylin-Eosin, Fluoro Jade B and immunohistochemically. The sections were then observed and evaluated under a light microscope. Results: Following the onset of hypoxic-ischemic encephalopathy in 7-day-old rat pups, damage to the investigated structures was observed in two animals. Other animals in the experimental group exhibited only minor morphological changes in neurons observable in H&E staining. Brains of the control group were intact. Keywords: necrosis,...

O modelo de recaída cue-induced é comumente utilizado para o estudo da recaída ao uso de drogas causada pela exposição dos animais à estímulos ambientais previamente associados à droga. Quando o álcool é a droga a ser estudada, um estímulo contextual (S) e um reforçador condicionado (Sr) são os estímulo frequentemente associados ao seu consumo. Embora esse modelo seja consolidado na área, sua validade de constructo vem sendo criticada; entre essas críticas estão o fato de que apenas o Sr adquire controle sobre a resposta de procura pela droga, além de que pouco se sabe sobre os sistemas neurais envolvidos nessa recaída. Portanto, este trabalho teve como objetivo principal avaliar o papel dos processos operantes no procedimento do modelo de recaída ao uso de álcool cue-induced, além de avaliar a participação de sistemas neurais ligados ao estresse nessa recaída. Ainda, este trabalho teve como objetivo melhorar o procedimento desse modelo animal. Os experimentos apresentados no Capítulo 1 tiveram como objetivo melhorar o controle exercido por S sobre a recaída, aprimorando o modelo, além de avaliar processos de aprendizagem associativa que ocorrem nesse modelo. Dois experimentos foram realizados para atingir esses objetivos. No primeiro, dois grupos de ratos foram treinados no modelo cue-induced, porém um grupo foi treinado sob o esquema de razão fixa 1 (FR1) e o outro sob o esquema de razão variável 5 (VR5). No segundo experimento, os ratos foram treinados nesse modelo, porém os estímulos S e Sr foram apresentados separadamente na fase de treino. Os resultados desses experimentos indicaram que o treino separado foi eficaz em melhorar o controle de S sobre a recaída e que a troca de esquema de reforço não foi eficaz para aumentar esse controle. Os resultados também mostraram que quando os estímulo são treinados em conjunto o Sr sombreia o S e quando eles são treinados separadamente e testados como um composto ocorre somação. Os experimentos apresentados no Capítulo 2 tiveram o objetivo de avaliar se eixos neurais ligados à resposta ao estresse estariam envolvidos na recaída mimetizada pelo modelo cue-induced. Ratos Marchigian Sardinian Preferring (msP) e Wistars foram treinados no modelo cue-induced e, na fase de testes, Antalarmin, Metirapona e corticosterona (CORT) foram injetados i.p. nos animais. Os resultados mostraram que o Antalarmin (dose 20 mg/kg) bloqueou a recaída em ambas as cepas, que a Metirapona (doses 50 mg/kg e 100 mg/kg) bloqueou a recaída nos ratos msP e apenas a dose de 100 mg/kg bloqueou a recaída nos ratos Wistars e que a CORT não teve efeito na em nenhuma das cepas. Esses resultados mostraram o procedimento do modelo de recaída cue-induced pode ser aprimorado principalmente no que tange ao controle do S sobre a recaída. Os resultados também mostraram que respostas ao estresse influenciam a recaída observada nesse modelo / The cue-induced relapse model is a model commonly used to study relapse caused by environmental stimuli. In its procedure, the environmental stimuli are usually a contextual stimulus (S) and a conditioned reinforcer (Sr). Although this model is extensively used its construct validity has been criticized. Between these critics are the fact that only the Sr controls the drug seeking responses because this stimulus overshadows the S, in addition to the fact that little is known about the neural systems that are involved in cue-induced relapse. Therefore, this work had as main purpose to evaluate operant process in the alcohol cue-induced relapse and evaluate the role of stress neural systems in this relapse. In adition, this work had also the purpose to improve the animal model procedure. The experiments presented in Chapter 1 had the purpose to increase the control exercised by S over relapse and to evaluate associative learned in this model procedure. Two experiments were conducted to achieve this goal. In the first one two groups of rats were trained in the cue-induced relapse model but one group was trained under the fixed ratio 1 (FR1) and the other under the variable ratio 5 schedule (VR5). In the second experiment, rats were trained in the cue-induced procedure, but the stimuli S and Sr were presented separately during the training phase. The results showed that separate training was effective in increase the S control over relapse and that changing the schedule of reinforcement was not effective in increase this control. Also, results showed that summation occurred when the stimuli are trained separately and tested as a compound. The experiments presented in Chapter 2 had the purpose to evaluate if neural axis related to organic stress responses are involved in cue-induced alcohol relapse. Also, it was investigated if the HPA is involved in relapse. Marchigian Sardinian Preferring (msP) rats and Wistars rats were trained in the cue-induced model and during the test phase Antalarmin, Metyrapone and corticosterone (CORT) were injected i.p. in the animals. The results showed that Antalarmin (dose 20 mg/kg) blocked relapse produced by environmental cues in both rats strains, Metyrapone (doses 50 mg/kg and 100 mg/kg) blocked relapse in msP rats and only dose 100 mg/kg blocked relapse in Wistar rats and CORT had no effect on relapse in both strains. These results showed that the procedure used in the cue-induced model can be improved mainly in regard to the control from S over relapse. The results also showed that stress responses influence relapse in this model

A realização deste estudo teve como objetivo a estimação de componentes de (co) variância e parâmetros genéticos das características: peso (PES18), perímetro escrotal (PE18), precocidade de acabamento (PREC), musculosidade (MUSC) e altura (ALT), mensuradas aos 18 meses de idade, ganho de peso da desmama ao sobreano (GP345) e idade ao primeiro parto (IPP) de bovinos Nelore, utilizando-se modelos multicaracterísticas. Foram avaliados dados de 107.332 mil bovinos criados entre os anos de 1994 e 2009 em fazendas localizadas nos estados de São Paulo, Mato Grosso do Sul e Goiás, e pertencentes ao Programa de Seleção da raça Nelore da Agropecuária CFM Ltda. Utilizaram-se três diferentes modelos: modelo multicaracterísticas padrão, modelo de componentes principais e de análise de fatores, esses dois últimos contemplando os três primeiros componentes e fatores, os quais foram comparados por critérios de informação de Akaike (AIC) e Bayesiano de Schwarz (BIC). O modelo que utilizou a técnica da análise de fatores contemplando os dois primeiros fatores (MFA2) apresentou melhor ajuste, seguido do modelo que contemplou os 3 primeiros fatores (MFA3). Estes possibilitaram estimativas próximas das realizadas pelos modelos multicaracterísticas padrão, às quais se mostraram parecidas com os valores já relatados e estimados no Brasil. / This study was objectified the estimation of (co)variance components and genetics parameters of traits: weight (W18), scrotal circumference (SC18), precocity finishing (PREC), muscularity (MUSC) and height (H), measured at 18 months of age, weight gain from weaning to yearling (WG345) and age at first calving (AFC) from Nellore cattle, using multi trait models, with analysis multi trait standard, analysis of principal components and factor analysis. Were evaluated data from 107,332 bovines, managed between the years of 1994 and 2009 in farms localized on states of São Paulo, Mato Grosso do Sul and Goias, and were participants in Nellore Selection Program from CFM Company Ltda. Was utilized three different models, multi trait model, model of principal components and factor analytic, these two last contemplating the four first principal components and factors analytics and that was compared by information criteria of Akaike (AIC) and Bayesian of Schwarz (BIC). The model that used the factor analysis technique contemplating the first two factors (MFA2) presented better adjustment, followed by the model that contemplated the first 3 factors (MFA3). These allowed close estimates to those performed by the standard multi trait models, which were similar to the values already reported and estimated in Brazil.

Chronic Mild Stress (CMS) é um modelo animal de depressão no qual ratos são submetidos a um protocolo de estressores moderados de forma crônica. Em função disso, o consumo e preferência por água com sacarose diminuem. Tal redução costuma ser considerada uma medida de anedonia, sintoma central da depressão em humanos. Três estudos realizados no Laboratório do Programa de Estudos Pós- Graduados em Psicologia Experimental da PUC-SP demonstraram que esse efeito é atenuado pela exposição a uma condição operante em esquema concorrente FR água FR sacarose. O presente estudo teve por objetivo investigar se a submissão a uma condição operante que envolvesse diferentes estímulos reforçadores teria o mesmo efeito. Para isso, as seguintes condições experimentais foram propostas: 1) exposição a um protocolo de estressores por seis semanas; 2) testes semanais de consumo de água e água + 2% sacarose; 3) sessões operantes. Dos nove sujeitos utilizados, quatroforam expostos também a uma condição na qual a resposta de pressão à barra foi consequenciada com acesso à roda de atividades, dois a uma condição na qual a resposta de pressão à barra foi consequenciada com uma pelota de alimento em FR6 e dois em FR12. Como resultado, observou-se que no sujeito submetido somente aos estressores (sujeito 09) foi replicado mais uma vez o efeito de redução no consumo de solução de sacarose. Esse efeito não pode ser observado em nenhum dos sujeitos submetidos às sessões operantes durante a exposição. Observou-se uma queda somente no primeiro teste após o término dessa, indicando que possivelmente a submissão a essas afetou o efeito tradicionalmente observado em decorrência da exposição ao protocolo de estressores. Tais resultados corroboram e ampliam os estudos anteriores. Nas sessões operantes, observou-se redução na taxa de respostas na quinta semana de exposição para os sujeitos em FR12 (02 e 03) e na quinta ou sexta para o sujeitos em FR6 (01 e 04). O desempenho na roda de atividades pareceu ficar sob controle da alteração no peso corporal (sujeitos 05 a 09). É possível que a exposição à condição operante tenha atenuado os efeitos do protocolo de estressores e a redução no valor reforçador dos estímulos tenha sido retardada e/ou observada em condições com maior custo de resposta. / Chronic Mild Stress (CMS) is an animal model of depression. Chronic exposure of rats to a protocol of mild stressors produces decrease in sucrose intake and reduction in the preference for sucrose over water, which is considered as a measure of anhedonia, a core symptom of depression. Three Brazilian studies from PUC-SP showed that the additional exposure of rats to operant sessions using FR water FR sucrose concurrent schedule of reinforcement attenuates this effect. The objective of this study was to investigate if the submission to operant conditions that involves other reinforcers would also attenuate the decrease in sucrose consumption. Three experimental conditions were proposed: 1) exposure to a protocol of mild stressors for six weeks; 2) weekly tests of water and water + 2% sucrose intake; 3) operant sessions. Subject 09 was exposed only to conditions (1) and (2). Eight subjects were submitted to conditions (1), (2) and (3): access to a running-wheel served as reinforcer, in CRF, for subjects 05, 06, 07 and 08 and food was the reinforcer for other four subjects: 01 and 04 were submitted to a FR6 schedule of reinforcement and 02 and 03 to a FR12 schedule of reinforcement. It was observed that the subject 09, that was submitted only to stressors showed, again, a decrease in sucrose consumption and preference. The other eight subjects did not show this effect. A decrease in sucrose consumption was noted only in the first test after exposure to the protocol. These results suggest that submission to these operant conditions affected the traditional effect of the exposure to mild stressors and it corroborates and amplifies previous studies. During operant conditions, it was observed that lever presses decreased during fifth week of exposure to stressors for subjects responding on FR12 and during fifth or sixth week for subjects responding on FR6. Performance on running wheel changed according to body weight. It is possible that exposure to these operant conditions diminished the stressors effects and that the decrease in the reinforcing value of these stimuli was delayed and/ or observed under conditions with higher response cost.

A psoríase é uma doença de pele inflamatória crônica, que afeta cerca de 2-4% da população mundial. Se desenvolve ao longo do tempo, principalmente no final da adolescência ou início da idade adulta e depende de uma complexa interação entre fatores genéticos e ambientais. Dados experimentais demostram que a dermatite induzida por imiquimode (IMQ) em camundongos assemelha-se estreitamente às lesões de psoríase humana, tanto nas características fenotípicas e histológicas como no desenvolvimento das lesões na epiderme. O estudo avaliou os efeitos anti-inflamatórios do óleo de semente de maracujá (Passiflora edulis) no tratamento da psoríase, utilizando a análise histológica e imunológica da epiderme. O experimento foi realizado com 36 camundongos Balb/c, os quais foram submetidos à indução da psoríase por imiquimode, por 10 dias consecutivos. O tratamento foi realizado com óleo de semente de maracujá in natura 100%, pomada LECIGEL® 2%, pomada associação de óleo de semente de maracujá 20% e LECIGEL® 2%, por 15 dias. Tanto o óleo da semente do maracujá quanto a associação do mesmo ao LECIGEL® diminuíram o quadro inflamatório induzido por imiquimode nas orelhas tratadas. Através das análises imuno-histoquímicas realizadas na epiderme (PCNA, IL-6, VEGF, CD34), observou-se um aumento na formação de corpos apoptóticos, diminuição a hiperplasia epitelial e redução do infiltrado inflamatório. Os resultados deste experimento demonstraram que o óleo da semente do maracujá desempenha um efeito anti-inflamatório no tratamento da psoríase induzida por imiquimode. / Psoriasis is a chronic inflammatory skin disease that affects about 2-4% of the world\'s population. It develops over time, especially in late adolescence or early adulthood and depends on a complex interaction between genetic and environmental factors. Experimental data show that imiquimode-induced dermatitis (IMQ) in mice closely resembles human psoriasis lesions, both in phenotypic and histological characteristics and in the development of lesions in the epidermis. The study evaluated the anti-inflammatory effects of passion fruit (Passiflora edulis) oil in the treatment of psoriasis, using the histological and immunological analysis of the epidermis. The experiment was performed with 36 Balb / c mice, which were submitted to psoriasis induction by imiquimode, for 10 consecutive days. The treatment was carried out with 100% fresh passion fruit seed oil, LECIGEL ® 2% ointment, 20% passion fruit seed oil ointment and LECIGEL ® 2% for 15 days. Both passionflower seed oil and its association with LECIGEL ® decreased the imiquimod-induced inflammation in the treated ears. Through the immunohistochemical analyzes performed on the epidermis (PCNA, IL-6, VEGF, CD34), an increase in the formation of apoptotic bodies, decrease in epithelial hyperplasia and reduction of inflammatory infiltrate was observed. The results of this experiment suggest that passion fruit seed oil has an anti-inflammatory effect in the treatment of imiquimode-induced psoriasis.

Os objetivos desse trabalho foram avaliar in vitro a sensibilidade de zigotos murinos à Brucella abortus, assim como a eficácia das lavagens seqüenciais e do tratamento com tripsina, padronizados pela International Embryo Transfer Society, na sua remoção e/ou inativação, a fim de estabelecer um modelo acessível para o estudo de interações embriões-patógenos. Para a coleta dos zigotos, foram utilizados camundongos fêmeas (Swiss Webster), púberes, nulíparas, 6-8 semanas de idade, acasaladas com machos inteiros da mesma linhagem, após superestimulação ovariana. Para a infecção foi utilizada a bactéria B.abortus 1119.3. A suspensão de bactérias foi preparada no momento da inoculação, na diluição de 106 bactérias/ml. Os zigotos obtidos foram separados em controle e infectados. A infecção dos zigotos foi feita com 30&micro;l da suspensão de bactérias, e após 24h e 96h foram analisados quanto à morfologia e taxa de clivagem. Os procedimentos de lavagem seqüencial e tratamento com tripsina foram realizados após 24h. Para detecção da bactéria após os procedimentos de lavagem, as amostras foram inoculadas em cultura de bactérias e submetidas ao teste de reação em cadeia pela polimerase (PCR). Uma amostra da última gota de lavagem de cada grupo também foi testada. Para analise estatística dos resultados foi utilizado o teste do &chi;2. No grupo controle não foram verificadas alterações morfológicas, já no infectado foi possível verificar a presença de blastômeros irregulares, falha de divisão, citoplasma com granulação e aspecto degenerativo. As taxas de clivagem obtidas foram de 77,4% (controle) e 59,2% (infectados) (&chi;2 de 0,001674; p<0,05) após 24h e após 96h de infecção 14,5% (controle) e 7% (infectados) (&chi;2 de 0,141616; p<0,05). Não foi possível isolar B.abortus em cultura após os procedimentos de lavagem para ambos os grupos. As amostras do grupo controle apresentaram-se negativas na PCR. Já no grupo infectado foram obtidos resultados positivos e negativos, para os embriões e para a alíquota da última gota de lavagem dos grupos tratados com a lavagem seqüencial. Foi verificada apenas em uma amostra a presença de embriões positivos para a B.abortus com amostra da última gota de lavagem negativa. Para outras duas amostras os embriões apresentaram-se livres de B.abortus, mas foram detectadas na alíquota da última gota de lavagem. Apenas em uma amostra foram obtidos resultados negativos tanto para os embriões, quanto para o meio de lavagem. Os resultados da PCR para os grupos infectados tratados com tripsina foram positivos para praticamente todas as amostras, contendo embriões ou apenas da alíquota da última gota de lavagem, com exceção dos embriões de uma amostra que apresentaram-se negativos. As novas tecnologias desenvolvidas em reprodução animal promovem manipulação invasiva do embrião. Portanto, tratamentos preconizados para a remoção de patógenos podem não ser eficazes. O risco a ser considerado é da presença de patógenos associados à zona pelúcida ou nas proximidades, que possam ser introduzidos, ou ter sua entrada facilitada no embrião. Considerando os dados apresentados, torna-se clara a importância do desenvolvimento de um modelo animal para estudos de interações embriões- patógenos, a fim de se evitar a transmissão e disseminação de doenças. / The aim of this study was evaluate in vitro mouse zygotes sensitivity to Brucella abortus, such as the embryo washing procedure and trypsin treatment, recommended by the International Embryo Transfer Society, efficacy in its removal and/or inactivation, with the purpose of established an accessible model for embryo- pathogen interaction studies. The female mice (Balb C) aging 6 to 8 weeks were superovulated and matted with fertile males of the same strain, then the zygote retrieval was performed. The bacterial suspension was prepared in the moment of inoculation and the dilution was 106 brucellas/ml. The zygotes were divided in control and infected groups (30&micro;l of the bacterial suspension); after 24h and 96h their morphology and viability were analised. The zygotes of each group were washed sequentially or treated with trypsin after 24h exposition. To verify the presence of B. abortus pos-washing the zygotes and a sample of the last wash drop of each group were tested on bacterial culture system and polimerase chain reaction (PCR). The statistical analyses was performed with the &chi;2 test. Morphological changes were not observed at the control group, but the infected one presented irregular blastomeres, clivage defective, granular cytoplasm with degenerative like morphology. The clivage rates were 77,4% (control) and 59,2% (infected) (&chi;2 de 0,001674; p<0,05) after 24h and after 96h 14,5% (control) e 7% (infected) (&chi;2 de 0,141616; p<0,05). Our results showed that the bacterial culture presented negative growing for all groups tested. The control group presented only negative results on PCR analyses. The infected group presented positive and negative results on PCR, for embryos or last wash drop sample, submitted to the sequential washing procedure. Positive embryos were found in just one sample with the last wash drop negative. To two other samples embryos presented negative PCR results but positive at the last wash drop. Negative results for embryos and last wash drop was found in just one sample. The PCR results for the groups treated with trypsin were almost all positives, for embryos or last wash drop samples, except one embryo sample that presented negative. The recent developed reproductive technologies promote excessive manipulation of the embryo. Therefore, preexistent procedures for microorganisms\' removal could not be efficient. The preeminent risk to be considered here is the potential or probability of the presence of pathogens associated with or in proximity to the zona pelúcida, which could be introduced, or facilitated its entrance on embryo. According to the data presented become clear the importance of develop a model for studies of embryo- pathogen interactions, with the aim of avoid disease transmission.

Foi desenvolvido um modelo experimental de Acidente Vascular Encéfalo Isquêmico (AVE) em suínos utilizando seis animais jovens, oriundos de granjas da região da Grande São Paulo, com 20kg de peso vivo em média, sendo todas fêmeas e hígidas. Estes foram submetidos a sedação com Quetamina (5mg/kg), Midazolan (0,5mg/kg) e Fentanil (0,005mg/kg) por via intramuscular e posteriormente intubados via oro - traqueal. A manutenção anestésica foi realizada utilizando isoflurano (vaporizador calibrado a 1,5%). Puncionou-se uma veia auricular para fluido e medicação de emergência se necessário. Os animais em plano anestésico foram submetidos a uma cateterização da artéria femoral que com auxilio de um fluoroscópio (Philips). Guiouse um cateter até a artéria carótida interna esquerda, local onde foi identificada a rede admirável epidural rostral. Mediante injeção de contraste (Telebrix), constatou-se que a cateterização apresentava-se seletiva na artéria carótida interna em posição ventral à rede admirável. Com esta confirmação fez-se a infusão de 1,0g de micro esferas de vidros que possuíam aproximadamente 0,4mm de diâmetro. Este procedimento de seletivação da artéria carótida interna foi repetido no antímero direito, realizandose a infusão de mesma quantidade de micro esferas, perfazendo um total de 2g de micro esferas de vidros por animal. Após 4 dias (96 horas) estes animais foram submetidos ao exame de ressonância nuclear magnética no Instituto de Física da USP. Para este procedimento os animais eram mantidos anestesiados com propofol (3mg/kg) por infusão continua durante todo o exame. A análise das imagens obtidas pela ressonância nuclear magnética indicou que este procedimento causou pontos de isquemia cerebral principalmente no antímero esquerda do cerebelo dos animais. Este fato confirmou as observações realizadas pela sintomatologia clínica apresentada pelos animais nos dias que antecederam o exame de ressonância nuclear magnética. Com estas lesões podemos afirmar que a metodologia desenvolvida para reprodução de AVE em suínos foi estabelecida. O modelo permite realizar estudos terapêuticos para esta doença que acomete um significativa parcela da população economicamente ativa da sociedade brasileira. / An experimental model of ischemic stroke (AVE) was investigated in healthy young female swine, at 20kg weight, of farms of São Paulo. The animals were sedated using an association of ketamine (5m/kg), midazolam (0.5mg/kg) and Fentanyl (0.005 mg/kg) IM; the maintenance was provided by tracheal entubation and isofluorane (1.5%). Fluid therapy was promoted through a catheter displaced in an auricular vein. Followed the catheterization of the femoral artery with the aid of a fluoroscope (Phillips®), the catheter was directed to the left internal carotid artery being possible the identification of the rete mirabile epidurale rostrale with the application of a radiopaque contratst. After the identification of the rete mirabile epidurale rostrale it was promoted the infusion of 1.0g of glass microsphere (0.4mm diameter); the same procedure was promoted on the right antimere through the right internal carotid artery, therefore, 2.0g of glass microsphere was injected in each animal. The animals were submitted to resonance scanning exam at day 4 (96 hours), after the surgical event, at the Physics Institute - USP. The resonance exam was carried out with the animals under intravenous anesthesia using continuous infusion of propofol (3mg/kg). The scanned images were analyzed and revealed focal cerebral ischemia at some regions mainly in the left antimere of the cerebellum, which can explain the clinical symptoms. We would infer the applied technique is suitable to develop ischemic stroke in swine and the animal modeling allows the investigation of therapeutics for this illness which compromises a significant economically.