La maladie d’Alzheimer est la maladie neurodégénérative la plus fréquente chez l’Homme et constitue un enjeu économique et de santé publique majeur. Les cerveaux de patients atteints présentent une atrophie corticale associée à deux types de lésions : les dégénérescences neurofibrillaires, constituées de protéines Tau agrégées, et les plaques amyloïdes, composées majoritairement de peptides amyloïde beta Aß agrégés. Les peptides Aß et leur agrégation seraient à l’origine de la pathogenèse. Afin d’éclaircir les mécanismes moléculaires à la base de la toxicité d’Aß, nous avons construit un modèle de toxicité d’Aß dans la levure Saccharomyces cerevisiae. Ce modèle a permis d’établir que la toxicité d’Aß dans la levure est intimement liée à sa sécrétion et au trafic vésiculaire. Ce modèle nous a également permis de réaliser une étude structure-toxicité du peptide et de mettre en évidence des éléments en cis importants pour la toxicité d’Aß. Une nouvelle voie d’agrégation des peptides toxiques en structure riche en feuillet ß anti-parallèle a pu ainsi être mise en évidence. Le modèle de toxicité d’Aß et l’existence de variants très toxiques d’Aß dans la levure nous a permis de réaliser des cribles génétiques afin de rechercher les éléments modulant la toxicité d’Aß in vivo. Le trafic vésiculaire, en particulier l’endocytose via le remodelage du cytosquelette d’actine, un complexe responsable de la formation de vésicules intraluminales appelé ESCRT, forment autant de pistes à étudier pour améliorer notre compréhension de la toxicité d’Aß. / Alzheimer’s disease is the most common neurodegenerative disease. This pathology is caused by aggregation of Aß peptides. The exact mechanism of neuronal cell dysfunction in Alzheimer’s disease is poorly understood and numerous models have been used to decipher the mechanisms leading to cellular death. In order to clarify the molecular mechanisms underlying the toxicity of Aß, we generated a new model to study Aß toxicity in yeast Saccharomyces cerevisiae. In our model, Aß toxicity is closely related to its secretion and its intracellular traffic. Indeed, when Aß is targeted to the secretory pathway, it is able to produce toxic species. Interestingly, we demonstrated also that even if Aß is addressed to the secretory pathway, it is still able to form cytoplasmic aggregates. Moreover, with this model, we generated new highly toxic mutants of Aß by random mutagenesis. In order to correlate structural conformation ‘signature’ to Aß toxicity, we performed a structure-toxicity study of these new variants. In vitro, we demonstrated that a new anti-parallel aggregation pathway is associated with highly toxic mutants of Aß. Then, using our Aß yeast model and also these harmful variants, we performed genetic screens in order to identify candidate genes able to modulate Aß toxicity in vivo. Given these different screens, we found that vesicular trafficking, endocytosis via actin cytoskeleton remodeling, and ESCRT-III (Endosomal Sorting Complex Required for Tansport) open new avenues to improve our understanding of Aß toxicity.

Introdução: A quimiorradioterapia (QRT) concomitante baseada em cisplatina é uma opção de tratamento empregada para os pacientes com carcinoma epidermoide de cabeça e pescoço (CECCP) localmente avançado e com bom performance status, seja em caráter adjuvante ou definitivo. O ganho de sobrevida com esta modalidade de tratamento é acompanhado de aumento das toxicidades agudas em comparação com a radioterapia isolada. A ocorrência de toxicidades tardias é menos reportada na literatura e incluem xerostomia, disfagia, hipotireoidismo, ototoxicidade, fístula/necrose cutânea, dentre outras. Tais sequelas tardias podem comprometer a qualidade de vida do sobrevivente ao CECCP. Objetivos: Verificar a prevalência de toxicidades tardias em sobreviventes ao CECCP tratados com QRT baseada em cisplatina. Métodos: Estudo transversal, uni-institucional, que incluiu de forma sequencial pacientes acima de 18 anos, tratados para CECCP (sítios primários: nasofaringe, orofaringe, cavidade oral, hipofaringe e laringe) e que haviam recebido QRT adjuvante ou definitiva, baseada em cisplatina. Estes pacientes estavam em seguimento há pelo menos 2 anos, sem evidência de doença. Os pacientes realizaram audiometria, endoscopia digestiva alta (EDA), nasofibrolaringoscopia da deglutição (NFL), exames laboratoriais (toxicidade tireoidiana e renal). Os pacientes incluídos também foram examinados clinicamente e as toxicidades apresentadas foram graduadas de acordo com a escala de toxicidades tardias do RTOG/EORTC. Os sobreviventes foram ainda avaliados quanto à percepção das toxicidades através de um inventário de sintomas e responderam questionários de qualidade de vida. Resultados: De janeiro de 2014 a fevereiro de 2017, 120 pacientes assinaram o TCLE. A idade mediana dos pacientes é 59 anos (21-78), com predomínio do sexo masculino (73%) e da cor branca (58%). Antecedente de tabagismo foi referido por 80% da amostra e de etilismo por 63%. Referente ao sítio primário, a maioria dos pacientes apresenta tumor em orofaringe (42%), seguido por laringe (23%) e cavidade oral (19%). O tempo de seguimento mediano é 42 meses (24-125). Há predomínio de pacientes com doença localmente avançada, tumores T3/T4 em 75% da amostra e N+ em 72%. A dose mediana de cisplatina recebida durante a concomitância foi 300 mg/m² (100-300) e de radioterapia foi 70 Gy (60-70,4). A QRT foi oferecida em caráter adjuvante em 49% da amostra. As toxicidades mais relatadas pelos pacientes foram: xerostomia (83%), alteração na voz (74%), saliva pegajosa (73%) e disfagia (73%). Ao se graduar as toxicidades conforme escala do RTOG/EORTC, verificou-se que a maioria das toxicidades apresentadas eram de graus leves, 1 ou 2. EDA encontrou estenose faríngea em 10% dos pacientes e NFL identificou fibrose em 37% dos sobreviventes. Dos pacientes submetidos a audiometria, 42% apresentaram perda auditiva de possível causa ototóxica. Cerca de 14% dos sobreviventes apresentam clearance de creatinina estimado < 60 mL/min/1,73m². Conclusões: Toxicidades tardias foram frequentemente reportadas pelos sobreviventes ao CECCP após QRT, porém, na maioria das vezes, de intensidade leve (graus 1 ou 2). Após a QRT, um seguimento cuidadoso é essencial para diagnóstico precoce e reabilitação a essas toxicidades, a fim de preservar a funcionalidade e qualidade de vida dos pacientes / Background: Cisplatin based CRT is the standard therapy for patients with locally advanced HNSCC with good performance status either as adjuvant or as definitive treatment. The survival gain with this treatment modality is accompanied by an increase in acute toxicities in comparison with isolated radiotherapy. The occurrence of LT is less reported in the literature and includes xerostomia, dysphagia, hypothyroidism, ototoxicity, cutaneous fistula / necrosis, among others. Such late sequelae may compromise the survivor\'s quality of life. Endpoints: To verify the prevalence of late toxicities in HNSCC survivors treated with cisplatin based CRT. Methods: A cross-sectional study that sequentially included patients over 18 years of age who were previously treated for HNSCC (primary sites: nasopharynx, oropharynx, oral cavity, hypopharynx and larynx) and who had received either adjuvant or definitive cisplatin based CRT. These patients were in follow-up for at least 2 years, with no evidence of disease. The patients underwent audiometry, upper GI endoscopy, nasopharyngolaryngoscopy (NPL), laboratory tests (thyroid and kidney toxicity). The included patients were also clinically assessed for mucous membrane, skin, subcutaneous tissue, salivary gland, larynx and esophagus LT according to the RTOG/EORTC Late Radiation Morbidity Scoring Schema. All patients answered a questionnaire about their perception of LT through a symptoms inventory and also answered QoL questionnaires. Results: From January 2014 to February 2017, 120 patients signed the informed consent form. The mean age of the patients is 59 years (21-78), predominantly male (73%) and white (58%). Previous smoking habits were reported by 80% of the sample and alcohol consumption by 63%. Most common primary sites were oropharynx (42%), followed by larynx (23%) and oral cavity (19%). The median follow-up time is 42 months (24-125). There was a predominance of locally advanced disease, T3 / T4 tumors in 75% of the sample and N + in 72%. The median cisplatin dose during concomitance was 300 mg/m² (100-300) and the median radiotherapy delivered dose was 70Gy (60-70.4). CRT was delivered as an adjuvant treatment in 49% of the sample. The most frequently selfreported LT were xerostomia (83%), voice disorders (74%), sticky saliva (73%) and dysphagia (73%). Assessing the toxicities according to the RTOG / EORTC scale most of them were mild, grade 1-2. Upper GI endoscopy diagnosed stenosis in 10% of the patients and NPL identified fibrosis in 37% of the survivors. Audiometry identified ototoxic hearing loss in 42% of the sample. About 14% of the survivors present chronic kidney disease (an estimated creatinine clearance < 60 mL/min/1.73m²). Conclusion: High rates of self-reported LT were detected although most of them seem to be mild. After CRT, a close follow-up of HNSCC patients is essential for early diagnosis, treatment of these late sequelae and rehabilitation, in order to preserve QoL and functionality and to avoid lifethreatening conditions and social reclusion

Espécies reativas de oxigênio (EROs) são produzidas normalmente durante o metabolismo de organismos aeróbios. Fungos degradadores de lignina geram estas espécies também no processo de degradação da lignina. As espécies de fungos bioluminescentes Gerronema viridilucens e Mycena lucentipes foram utilizadas para se estabelecer possível dependência entre intensidade de bioluminescência, viabilidade celular e atividades das enzimas de defesa antioxidante e ligninolíticas nos micélios e corpos de frutificação. Verificou-se o efeito de espécies causadoras de estresse químico (metais e fenóis) na emissão de luz, viabilidade celular, defesas antioxidantes e enzimas de respiração celular no fungo bioluminescente G. viridilucens, com o objetivo de conectar a inibição da bioluminescência com danos oxidativos ao organismo. Constatou-se que diferentes espécies de fungos bioluminescentes podem apresentar características diferentes quanto à emissão de luz e proteção antioxidante. Diferenças na intensidade e variação temporal da emissão de luz de diferentes espécies foram observadas nos corpos de frutificação e também no micélio. Isto foi revelado pela irregularidade do perfil de luz reprodutível para a espécie M. lucentipes, ao contrário do observado para G. viridilucens. A viabilidade celular de ambas as espécies varia com o tempo, sendo que no caso de G. viridilucens seu perfil é similar ao da bioluminescência. Os ensaios enzimáticos indicam maior atividade no micélio dos fungos do que nos corpos de frutificação, provavelmente pela função específica reprodutora do corpo de frutificação, enquanto a atividade metabólica do fungo está concentrada no micélio. As enzimas ligninolíticas também apresentam atividade baixa nas culturas estudadas, provavelmente por serem enzimas de degradação extracelulares. O fato de ambas viabilidade celular e bioluminescência do micélio serem reduzidas na presença dos metais (cobre e cádmio) e fenóis (fenol e 2,4,6-triclorofenol) testados atesta a interrelação entre atividade luminogênica e injúria oxidativa aos fungos. Os metais parecem afetar mais negativamente as defesas antioxidantes dos fungos do que os fenóis, os quais possivelmente são eliminados pela atividade da glutationa S-transferase (GST), sem também afetar as demais defesas antioxidantes. No conjunto, estes resultados possibilitam estabelecer uma relação metabólica entre abatimento da bioluminescência e as defesas antioxidantes do organismo. No caso dos metais, os sistemas de defesa antioxidante envolvendo a glutationa são bastante importantes, tanto para eliminar peróxidos produzidos na presença de cobre, como na quelação de cádmio pela glutationa. Sob condições normais, a bioluminescência, defesas antioxidantes e respiração celular do organismo estariam funcionando e o NAD(P)H seria mobilizado por todos estes sistemas. Quando o fungo é submetido ao estresse químico, o fluxo de NAD(P)H seria desviado da bioluminescência para sustentar prioritariamente as defesas antioxidantes e a respiração celular, essenciais para a proteção, manutenção e reprodução do organismo / Reactive Oxygen Species (ROS) are normally produced during the metabolism of aerobic organisms. Ligninolitic fungi also produce these oxidizing species also during the lignin degradation process. The bioluminescent species Gerronema viridilucens and Mycena lucentipes were studied aiming to establish a correlation between the temporal profiles of bioluminescence, cellular viability and antioxidant defense enzymes and ligninolitic enzymes in mycelium and fruiting bodies. Chemical toxicants such as metals and phenols were here found to affect light emission, cellular viability, antioxidant defenses and cellular respiration enzymes when administered to G. viridilucens, thereby attesting a metabolic connection between bioluminescence inhibition and fungal oxidative damage. Different species of fungi exhibit different characteristics linked to light emission and antioxidant defenses. Differences in light emission displayed by different species do not resume to fruiting bodies, but the light profile and intensity can also vary in the mycelia. This may explain the irreproducibility of the light profile from M. lucentipes, differently to that observed with G. viridilucens. The cellular viability of both species varies with time, G. viridilucens profile being similar to the time course of bioluminescence. The enzymatic data pointed to higher activities in mycelium than in fruiting bodies, probably due to a main reproductive function of the latter, whereas the metabolic activities are prevalent in the mycelium. Ligninolytic enzymes exhibit low activities in the extracts of fungus samples, probably because they are extracellular degradation enzymes. The inhibition effect of phenols and metals (copper and cadmium) on mycelium viability reinforces the notion that cellular oxidative damage hampers bioluminescence emission. Redox active (copper) and heavy metals (cadmium) were found to display higher impact on antioxidant defenses than phenols (phenol and 2,4,6-trichlorophenol), which are expected to be promptly metabolized and excluded by principally glutathione S-transferase (GST). Notably, a metabolic correlation between bioluminescence inhibition and antioxidant defenses was unveiled by the present work. Regarding the metals, glutathione was found to be a crucial antioxidant, both to eliminate peroxides when in the presence of copper, and to act as a cadmium chelating agent. Under normal conditions, the bioluminescence system, the antioxidant defenses and the cellular respiration sets cooperate through the common demand of reducing power of NAD(P)H. Under chemical stress, the NAD(P)H flux would be deviated from bioluminescence, to principally sustain the antioxidant defenses and cellular respiration, essential for the protection, maintenance and reproduction of the organism.

Este estudo foi realizado na safra 2002/2003 quando foi avaliado os níveis de endosulfan e seus principais produtos de transformação numa área agrícola localizada na cabeceira de uma microbacia do município de Aguaí-SP. Os objetivos específicos foram: (i) avaliar a movimentação do endosulfan e seus principais transformação no solo sob a cultura do algodão (áreas submetidas à aplicação), (ii) avaliar sua transferência de compartimento (transporte para o solo sob milho e braquiária, adjacentes às áreas aplicadas, e para os corpos aquosos - nascente, lago, córrego jusante e poço), (iii) avaliar os resíduos de endosulfan e seus produtos de transformação nos peixes do açude e (iv) predizer, através de modelagem matemática, a concentração de endosulfan passível de atingir o corpo d\'água e a largura mínima da faixa de contenção (mata ciliar) para impedir a chegada dos produtos no lago. Para tanto, foram realizadas amostragens de solo, sedimento, água dos corpos hídricos e da enxurrada e de peixes, durante sete meses, nas quais foram realizadas análises de &#945;- e &#946;-endosulfan, endosulfan sulfato e endosulfan diol. As análises foram efetuadas por cromatografia gasosa com detecção por espectrometria de massa. Para prever a concentração do produto passível de atingir o açude, tanto associada ao sedimento como na forma livre, foi realizada uma modelagem matemática com base nas propriedades físico-químicas do produto e nas condições edafoclimáticas e de manejo da área estudada. Pôde-se concluir que: (i) o endosulfan e seu produto de transformação endosulfan sulfato não apresentam potencial de lixiviação no solo nas condições estudadas; (ii) a persistência do endosulfan no campo é menor em relação aos estudos de laboratório, sendo que o isômero alfa é o que apresenta a mais rápida degradação e o principal produto de transformação do endosulfan no solo é o endosulfan sulfato; (iii) a principal via de transporte do endosulfan e do endosulfan sulfato no ambiente é o escoamento superficial; (iv) o endosulfan e seus produtos de transformação são transportados associados as micropartículas de solo (\"resíduo-ligado\"); (v) quando a aplicação segue as recomendações estabelecidas pelo fabricante, o produto somente apresenta potencial para atingir áreas adjacentes se houver caminhamento da enxurrada para estas áreas; (vii) o endosulfan e seus produtos de degradação, em sua fase de \"resíduo-ligado\", não apresentam efeito agudo e bioacumulação para as espécies de lambari (Astyanax sp.) e tilápia (Oreochroms sp.) da área nas concentrações observadas neste estudo e (viii) pela modelagem matemática, dentro das condições estudadas, 40 metros de faixa de contenção seria suficiente para impedir a chegada do endosulfan e do endosulfan sulfato nos corpos d\'água, desde que não houvesse fluxo preferencial de enxurrada. / The study was carried out during 2002/2003, and its objective was to evaluate the levels of endosulfan and its main transformation products in a fanning area located at the head of a small valley close to the town of Águaí, SP, Brazil. The specific objectives were: (i) to evaluate the dynamics of endosulfan and its main transformation products in the soil beneath a crop of cotton (areas treated with endosulfan), (ii) to evaluate the transfer of endosulfan among various locations [transport from the applied areas into the adjacent areas cultivated with maize (Zea mays) and brachiaria (Brachiaria spp.), and into various bodies of water - a spring, a lake (pond), a small flowing stream and a well], (iii) to evaluate the presence of the residues of endosulfan and its transformation products on fish in the lake and (iv) to predict, using mathematical modeling, the endosulfan concentration able to reach the lake and the minimum width buffer zone (containment strip or riparian vegetation) needed to avoid the arrival of the pesticide in the lake. In order to achieve this, representative samples of the following types were collected - soil, sediment, fish, water from the spring, the pond, the stream and the well, as well as from the runoff. Samples were collected during a period of seven months, and were analyzed for &#945;- and &#946;-endosulfan, endosulfan sulfate and endosulfan diol. The analyses were performed by gas chromatography, with mass spectrometric detection. Mathematical approach was employed to predict the concentration of endosulfan that could reach the lake, and in which form it would be present (free or sorbed to soil microparticles). Input data for the model were the physical-chemical properties of endosulfan, the local soil and climatic conditions, and the management of the study area. It was possible to conclude that: (i) endosulfan and its transformation products endosulfan sulfate do not present potential leaching in the soil profile; (ii) the persistence of endosulfan in the field is shorter than has been found in laboratory studies, &#945;-endosulfan is degrades more rapidly than &#946;-endosulfan and endosulfan sulfate is the main transformation product; (iii) runoff is the principal transport mechanism for endosulfan and endosulfan sulfate in the environment; (iv) the transformation products are transported by attachment to microparticles of soil (\"bound residues\"); (v) when the application of endosulfan follows the recommendations of the manufacturer, the compound is only likely to reach adjacent areas if there is a runoff into these areas; (vii) the products, in the \"bound residue\" form, do not produce acute effects and bioaccumulation in the species of fish lambari (Astyanax sp.) and tilapia (Oreochroms sp.) from the area, and (viii) the modeling has established that, within the conditions of the study, a 40 m wide containment strip is sufficient to prevent the appearance of endosulfan and endosulfan sulfate in bodies of water, considering that there is no prefential flow of runoff water.

A planta Casearia sylvestris, popularmente conhecida como guaçatonga, é comumente utilizada para o tratamento de diversas afecções como queimaduras, ferimentos, herpes labial e genital, distúrbios do trato gastrointestinal, como gastrites, úlceras, gengivites, aftas, halitose; além disto, atribui-se à planta ação cicatrizante, tônica, depurativa, anti-reumática, antiinflamatória, antiofídica, antidiarréica, entre outras. Atualmente, é catalogada como planta medicinal de interesse ao Sistema Único de Saúde no Brasil. Entretanto, até o momento, não haviam estudos relativos à avaliação de sua toxicidade. Assim, o objetivo desta pesquisa foi o de estudar, em ratos, os possíveis efeitos tóxicos da C. sylvestris. Para tal, foram realizados os testes de citotoxicidade com o objetivo de analisar o potencial tóxico das diferentes formas de extração da planta, e estudos pré-clínicos de toxicidade oral aguda, e em doses repetidas, por 28 e 90 dias, realizados de acordo com protocolos preconizados pelas agências reguladoras nacionais (Agência Nacional de Vigilância Sanitária - ANVISA) e internacionais (Organization for Economic Cooperation and Development - OECD) de avaliação de risco para substâncias químicas. No presente estudo, o extrato fluido (EF) de C. sylvestris foi administrado, a ratos, por via oral, através de gavagem, em dose única de 2000mg/kg; ou em doses repetidas, durante 28 ou 90 dias, nas doses 60, 120 e 240mg/kg,. Foram avaliados sinais sistêmicos de toxicidade, ganho de peso e consumo de ração, hemograma completo, avaliação bioquímica (AST, ALT, GGT, ALP, uréia, creatinina, ácido úrico, colesterol, triglicerídes, glicose, albumina, proteínas totais e cloreto), análise macroscópica e histopatológica de órgãos e avaliação do peso relativo dos órgãos linfóides baço e timo e celularidades do baço e medula óssea para averiguar os possíveis efeitos imunotóxicos. Além disso, foi obtido o perfil cromatográfico (fingerprint) utilizando a técnica analítica CLAE/UV. Os resultados mostraram que o EF de C. sylvestris não causou efeitos tóxicos quando administrado em dose única 2000mg/kg no estudo de toxicidade aguda, assim como também não mostrou efeitos sistêmicos de toxicidade ou efeitos imunotóxicos nos estudos em doses repetidas por 28 e 90 dias. Estes dados aqui encontrados permitem sugerir que o EF da C.sylvestris é seguro quando utilizado mesmo prolongadamente. No entanto, outros estudos, como por exemplo, de genotoxicidade e de carcinogenicidade, devem ser realizados para complementar esta etapa de avaliação pré-clínica. / Casearia sylvestris Sw commonly known in Brazil as guaçatonga, is currently used to treat many diseases such as burns, wounds, labial and genital herpes, gastrointestinal disorders- gastritis, ulcers, gingivitis, aphthae and halitoses. C.sylvestris has been also used as cicatrizant in skin diseases, blood purifier and general detoxification, anti-inflammatory, antiofidic, diarrhea, among others. In our country it is considered as medicinal plant and for this reason C.sylvestris is catalogued at The Unified Health System (Sistema Único de Saúde - SUS) as a plant of interest for Brazilian population .However, no toxicological studies concerned the safety of extract fluid of this plant have been reported. Thus, the aim of this study was to evaluate the possible toxic effects of fluid extract (EF) of C. sylvestris. For this, it was performed citotoxicity tests in order to analyze the toxic potential of different extracts of the plant. It was also performed preclinical studies, in rats, according the guidelines proposed by National Health Surveillance Agency (Agência Nacional de Vigilância Sanitária - ANVISA) and OECD (Organization for Economic Cooperation and Development) being performed the following tests: acute oral toxicity and repeated dose 28 and 90-day. The EF of C. sylvestris was administered by gavage at a single dose of 2000 mg/kg, or during 28 and 90 days with doses 60, 120 e 240mg/kg. It was evaluated systemic signs of toxicity, body weights and food consumption, hematology, serum biochemistry (AST, ALT, GGT, ALP, urea: creatinine, uric acid cholesterol, triglycerides, glucose, albumin, total proteins, and chloride) macroscopic and histopathological analysis of different organs and evaluation of relative weight of lymphoid organs, spleen and thymus and the cellularity of spleen and bone marrow to investigated the possible immunotoxic effects. We also obtained the fingerprint chromatogram of C. sylvestris, using CLAE/UV. The results showed that there were no toxic effects by the EF when administered as a single dose at 2000mg/kg in acute toxicity study, as well as showed no systemic effects of toxicity or immunotoxic effects in studies with repeated doses of 28 and 90 days. This data suggest that the EF of the plant is safety even used for long period. However, other studies, eg, genotoxicity and carcinogenicity studies should be performed to complement the preclinical evaluation.

Defeitos ósseos metafisários grandes criados após curetagem de tumores ósseos são normalmente tratados com implantação de cimento sólido de polimetilmetacrilato (PMMA). Com os novos cimentos porosos, reabsorvíveis ou não, espera-se melhores resultados clínicos com diminuição da incidência de soltura asséptica relacionada à necrose térmica do osso, à discrepância das características mecânicas e à ausência de osteointegração. A mistura de componentes efervescentes, como o bicarbonato de sódio e o ácido cítrico, produz cimentos porosos cuja biocompatibilidade e segurança nunca foram testadas, limitando sua utilização clínica. O objetivo deste estudo foi avaliar a eliminação de substâncias tóxicas e o efeito anticoagulante do citrato de sódio, um subproduto da reação. Em experimentos in vitro o cimento sólido de PMMA foi comparado aos cimentos porosos de PMMA e poliuretana de mamona nas duas formas comercialmente disponíveis (Poliquil® e Bioosteo®). Todos os elementos produzidos durante a preparação e a incubação dos cimentos foram analisados por Cromatografia Líquida de Alta Eficiência acoplada a espectrometria de massa e por proliferação de culturas das linhagens celulares NIH/3T3 e MRC-5 coradas com resazurina. Em experimentos in vivo defeitos ósseos criados nos fêmures de coelhos não preenchidos foram comparados com outros preenchidos com os cimentos porosos à base de PMMA e Poliquil®. Foram realizados ressonância magnética, análise dos parâmetros de coagulação antes, imediatamente, 1, 4 e 7 dias após a cirurgia e análise histológica do membro operado, dos rins e do fígado dos coelhos. Ambas as marcas de poliuretana de mamona (Bioósteo® e Poliquil®) liberaram 4,4\'diaminodifenilmetano e a marca Poliquil® liberou também 4,4\'-difenilmetano diisocianato. Ambos os compostos são considerados tóxicos à inalação ou ingestão. A proliferação celular das culturas das linhagens MRC-5 e NIH/3T3 sofreu redução menor que 20% quando em contato com os cimentos à base de poliuretana de mamona e menor que 35% quando em contato com o cimento poroso à base de PMMA, em comparação com o grupo de controle. A diluição do meio de cultura diminuiu este efeito. Acreditamos que a acidez do meio, devido à reação incompleta dos elementos efervescentes, pode ser a causa deste efeito. A análise histológica do fígado e dos rins mostrou algumas pequenas alterações na mesma proporção que no grupo controle. Inferimos que os cimentos porosos não causam toxicidade aguda sistêmica. Em todos os grupos, a análise histológica da área operada mostrou pequenos hematomas e uma ligeira reação de corpo estranho. Não foi encontrada reação inflamatória importante em nenhum dos grupos. Estes dados corroboram a hipótese de que os cimentos porosos são biocompatíveis. O leve efeito citotóxico observado in vitro não pôde ser detectado in vivo, provavelmente pelas condições locais de diluição. A produção de citrato parece não interferir com a coagulação. Os parâmetros de coagulação não sofreram alterações em qualquer momento após a cirurgia e a avaliação por ressonância magnética, assim como a análise histológica, descartou a formação de hematomas consideráveis. Concluímos que o cimento poroso à base de PMMA testado é seguro para o uso clínico, mas que os cimentos porosos à base de poliuretana de mamona eliminam substâncias tóxicas cuja repercussão clínica merece investigação específica. / Large metaphyseal bone defects created after benign bone tumor curettage are usually treated by solid bone cement (PMMA) implantation. Porous and absorbable cements are expected to improve clinical results by lowering incidence of aseptic loosening, heat promoted bone necrosis, bone/cement mechanical discrepancies and absence of bone integration. Effervescent components like sodium bicarbonate and citric acid can be mixed to PMMA producing porous cement that fits intraoperative requirements. Furthermore, castor oil polyurethane is a well-known absorbable cement that can be mixed to effervescent components to produce porous and absorbable bone cement. In vivo and in vitro toxicity of both products have never been tested to support its clinical use. The spurious production of toxic elements and the effect of citrate, an anticoagulant byproduct of the effervescent components reaction, were analyzed. In vitro experiments: The three experimental groups consisted of PMMA or one of the commercially available castor oil polyurethane cements (Poliquil® or Bioosteo®) mixed to the effervescent components specimens. These groups were compared to classic PMMA solid cement specimens. All the elements produced during preparation and incubation of the cements were analyzed by High Efficiency Liquid Chromatography coupled to Mass Spectrometry and by Resazurine microplate assay of NIH/3T3 and MRC-5 fibroblasts strains culture. In vivo experiments: Femoral defects were created in six rabbits per group and filled with PMMA or Poliquil® porous cements or left empty. Magnetic resonance of the limb, histology of the limb, kidneys and liver, and coagulation parameters of blood samples collected immediately after and 1, 4 and 7 days after surgery were analyzed. Castor oil polyurethane of both brands (Bioósteo® and Poliquil®) released 4,4\'-diaminodiphenylmethane. The Poliquil® brand released 4,4\'-diphenylmethane diisocyanate too. Both compounds are considered toxic. The MRC-5 and NIH3T3 cell strains proliferation was decreased in less than 80% in contact with polyurethane cements and less than 65% with PMMA, compared to the control group. Dilution of the medium diminished this effect. We believe that the acidity of the medium due to incomplete reaction of the effervescent components may be the cause of this finding. Liver and kidneys histology showed some slight changes in the same proportion as the control group. We infer that the cements do not cause acute systemic toxicity. In all groups, histologic analysis of the operated area showed small hematomas and a slight foreign body reaction. Significant inflammatory reaction could not be found in any of the study groups. Citrate formation from the effervescent components reaction seems to not interfere with coagulation. Inflammatory reaction around porous cements is similar to that of classic solid PMMA cement. The cytotoxic effect observed in vitro, could not be detected in vivo. Coagulation parameters did not changed at any time after surgery. Magnetic resonance imaging evaluation confirmed that there was no formation of larger hematomas than in specimens of the control group. In conclusion, the tested porous PMMA cement is safe and biocompatible for clinical use. The tested polyurethane porous cements eliminates toxic substances and deserves specific studies to verify clinical safety.

Ecossistemas marinhos e estuarinos são o destino final de muitos contaminantes, e a ecotoxicologia ainda enfrenta a falta de organismos modelo para esses ambientes. Para desenvolver e apresentar um organismo teste adequado para ser utilizado em estudos de laboratório e campo, métodos para o cultivo, endpoints e biomarcadores devem ser avaliados. O anfípode marinho Parhyale hawaiensis é um modelo para estudos de biologia evolutiva e embrionária, é mundialmente distribuído e fácil de manipular em laboratório e consequentemente um modelo interessante para estudos ecotoxicológicos. O objetivo desse trabalho é padronizar condições de cultivo, teste de toxicidade, e desenvolver biomarcadores no anfípode P. hawaiensis e aplicá-los para avaliar os efeitos de nanopartículas metálicas na exposição via alimentação. As condições de cultivo foram estabelecidas em água salina reconstituída (salinidade 30 ± 2), temperatura de 24 ± 2 oC, fotoperíodo de 12-12h luz/escuro, coral triturado como substrato, alimentação diária com ração de peixe, troca de água parcial e aeração constante. As condições de teste compreendem a utilização de microplaca de 96-poços, organismos <7 dias, um organismo por poço com 200 &#181;L de solução por 96 h. Zinco, cobre, prata, cádmio e amônia foram selecionados como substância teste e a sensibilidade de P. hawaiensis foi similar e dentro da faixa de outros anfípodes marinhos. As condições para expressão gênica em P. hawaiensis foram desenvolvidas para o gene de referência 18S, dois genes de metalotioneínas (MT) e dois genes de glutationa S-transferase (GST). Entretanto, os genes selecionados não foram diferencialmente expressos nas condições testadas. Sendo assim, a análise da expressão gênica global, RNA-seq, foi realizada com organismos adultos alimentados por 7 dias com alimento contaminado com nanopartículas de prata (Ag-NPs), AgCl e controle. A análise da expressão gênica global (RNA-seq) foi conduzida com sucesso e destacou diferenças na resposta entre machos e fêmeas, bem como nos tratamentos com AgCl e Ag-NP. A análise comparativa revelou genes comuns entre as duas formas de Ag estudadas, e mostrou respostas diferentes no tratamento com Ag-NP, portanto, efeitos adversos diferentes são esperados após a exposição à alimentação, e os genes diferencialmente expressos podem ser usados como potenciais biomarcadores. Os resultados demonstraram que P. hawaiensis é um bom modelo para testes de toxicidade com um protocolo miniaturizado e o RNA-seq foi aplicado com sucesso para investigar as diferenças entre a exposição via alimentação com AgCl e Ag-NP e pode revelar potenciais biomarcadores. Além disso, este estudo desenvolveu protocolos e fornece informações moleculares que podem ser usadas em estudos futuros com P. hawaiensis. / Marine and estuarine ecosystems are the final destinations of many contaminants and ecotoxicology still faces the lack of model organisms for these environments. To develop and present a suitable test organism to be used in laboratory and field studies, methods for husbandry, ecotoxicity endpoints and biomarkers should be evaluated. The marine amphipod Parhyale hawaiensis is already a model for development and evolutionary studies, it is worldwide spread and easy to handle in the laboratory and so on an interesting model for ecotoxicology. The aim of this work is to standardize husbandry conditions, toxicity testing, and to develop molecular biomarkers in P. hawaiensis and apply them to evaluate the effect of metal nanoparticles in dietary exposure. The conditions for culturing were established in reconstituted seawater (30 ± 2 salinity), 24 ± 2 oC temperature, photoperiod 12-12 h light/dark, crushed coral as substrate, daily feeding with fish food, partial water exchange and constant aeration. Testing conditions comprised 96-well microplate, <7 days age organisms, one organism in each well containing 200 &#181;L of exposure media for 96 h. Zinc, copper, silver, cadmium and ammonia were selected as toxicants and sensitivity of P. hawaiensis were within the range of other marine amphipods. Gene expression conditions for P. hawaiensis were develop for the reference gene 18S, two metallothioneins (MT) and two glutathione-S-transferase (GST). However, selected target genes, analyzed by RT-qPCR, were not successful to detect differences in our treatment conditions. Thus, global gene expression analysis, RNA-seq, was conducted with adult organisms fed for 7 days to a contaminated food with silver nanoparticles (Ag-NPs), AgCl and control. Results highlighted differences between males and female\'s toxicity responses as well as AgCl and Ag-NP treatments. Comparison analysis revealed common genes expressed in both Ag forms, but also showed differences in Ag-NP treatment with both up and down regulated genes. Analysis are underway to better understand toxicity responses pathways. These results anticipate different responses to both Ag forms investigated, therefore different adverse effects are expected after feeding exposure to AgCl and Ag-NP and significative genes can be used as potential biomarkers. Our results demonstrated that P. hawaiensis is a good model for ecotoxicity tests with a miniaturized protocol and RNA-seq were successful applied to investigate the differences between AgCl and Ag-NP feeding exposure and could reveal promising biomarkers. Moreover, this study developed protocols and provide molecular information that can be used in future studies with P. hawaiensis.

by Wong Wing-yee, Felice. / Thesis (M.Phil.)--Chinese University of Hong Kong, 1998. / Includes bibliographical references (leaves 144-166). / Abstract also in Chinese. / Acknowledgements --- p.i / List of Abbreviations --- p.ii / Abstract --- p.iv / Abstract (Chinese Version) --- p.vi / Table of Contents --- p.viii / List of Tables --- p.xii / List of Figures --- p.xiv / List of Appendices --- p.xvi / Chapter Chapter I --- Literature Review / Chapter 1. --- Introduction --- p.1 / Chapter 2. --- Background of Cytochrome P450 --- p.3 / Chapter 2.1 --- Discovery --- p.3 / Chapter 2.2 --- Tissue Distribution --- p.3 / Chapter 2.3 --- Structure and Functions --- p.7 / Chapter 2.4 --- Nomenclature of the P450 Superfamily --- p.10 / Chapter 3. --- Cytochrome P450 2E1 (CYP2E1) --- p.11 / Chapter 3.1 --- Discovery --- p.11 / Chapter 3.2 --- Tissue Distribution --- p.12 / Chapter 3.3 --- Substrates and Inducers --- p.13 / Chapter 3.4 --- Toxicological Role of CYP2E1 --- p.15 / Chapter 4. --- CYP2E1-knockout Mouse Model --- p.17 / Chapter Chapter II --- Carbon Tetrachloride (CC14) Study / Chapter 1. --- Introduction --- p.19 / Chapter 1.1 --- General Properties and Usage of CC14 --- p.19 / Chapter 1.2 --- Toxicological Aspects of CC14 --- p.19 / Chapter 1.3 --- Mechanism of CCl4-induced Hepatotoxicity --- p.20 / Chapter 1.4 --- Role of CYP2E1 in CCl4-induced Hepatotoxicity --- p.23 / Chapter 1.5 --- Objectives of the Study --- p.27 / Chapter 2. --- Materials and Methods --- p.29 / Chapter 2.1 --- Chemicals and Materials --- p.29 / Chapter 2.2 --- Animals --- p.29 / Chapter 2.3 --- Acute CC14 Treatment --- p.29 / Chapter 2.4 --- Preparation of Microsomal Fractions --- p.30 / Chapter 2.5 --- Determination of Microsomal Protein Concentration --- p.31 / Chapter 2.6 --- Determination of Serum Aminotransferase Activities --- p.31 / Chapter 2.7 --- Liver Histology --- p.32 / Chapter 2.8 --- Hepatic Microsomal CYP2E1 Activity -p-nitrophenol Assay --- p.34 / Chapter 2.9 --- SDS-PAGE and Western Blot Analysis --- p.35 / Chapter 2.10 --- Detection of Lipid Peroxidation in vitro and in vivo --- p.35 / Chapter 2.10.1 --- In vitro Lipid Peroxidation - 2-Thiobarbituric Acid (TBA) assay --- p.35 / Chapter 2.10.2 --- In vivo Lipid Peroxidation - Microsomal Conjugated Dienes Detection --- p.36 / Chapter 2.11 --- Hepatic Lipid Fatty Acid Composition Analysis --- p.39 / Chapter 2.11.1 --- Lipid Extraction --- p.39 / Chapter 2.11.2 --- Thin Layer Chromatography --- p.39 / Chapter 2.11.3 --- Methylation --- p.40 / Chapter 2.11.4 --- Gas Chromatography --- p.40 / Chapter 2.12 --- Statistical Analysis --- p.41 / Chapter 3. --- Results --- p.42 / Chapter 3.1 --- "Mortality, Liver Weight and Liver Color" --- p.42 / Chapter 3.2 --- Hepatotoxicity --- p.42 / Chapter 3.2.1 --- Serum ALT and AST activities --- p.42 / Chapter 3.2.2 --- Liver Histology --- p.45 / Chapter 3.3 --- CYP2E1-catalysed PNP Activities and CYP2E1 Protein Levels --- p.49 / Chapter 3.3.1 --- CYP2El-catalyzed PNP Activities --- p.49 / Chapter 3.3.2 --- CYP2E1 Protein Levels --- p.52 / Chapter 3.4 --- Lipid Peroxidation --- p.52 / Chapter 3.4.1 --- In vitro Lipid Peroxidation --- p.52 / Chapter 3.4.2 --- In vivo Lipid Peroxidation --- p.54 / Chapter 3.5 --- Hepatic Lipid Fatty Acid Composition --- p.56 / Chapter 3.5.1 --- Fatty Acid Composition in Hepatic Phospholipid --- p.56 / Chapter 3.5.2 --- Fatty Acid Composition in Hepatic Microsomal Phospholipid --- p.59 / Chapter 3.5.3 --- Fatty Acid Composition in Hepatic Triglyceride --- p.61 / Chapter 4. --- Discussion --- p.63 / Chapter 4.1 --- CYP2E1 is Required in CCl4-mediated Hepatotoxicity --- p.63 / Chapter 4.2 --- CYP2E1 is Degraded following CC14 Exposure --- p.65 / Chapter 4.3 --- CYP2E1 is Required in CCl4-induced Lipid Peroxidation --- p.67 / Chapter 4.4 --- CYP2E1 is Required in CCl4-induced Hepatic Phospholipid Depletion --- p.70 / Chapter 4.5 --- CYP2E1 is Required in CCl4-induced Hepatic Triglyceride Accumulation --- p.72 / Chapter 5. --- Conclusion --- p.76 / Chapter Chapter III --- Chronic Ethanol Consumption Study / Chapter 1. --- Introduction --- p.77 / Chapter 1.1 --- Multiple Metabolic Pathways for Ethanol Metabolism --- p.77 / Chapter 1.2 --- Metabolism of Ethanol by the Microsomal Ethanol Oxidizing System --- p.79 / Chapter 1.3 --- Role of CYP2E1 in Ethanol Metabolism --- p.82 / Chapter 1.4 --- Role of CYP2E1 in Alcoholic Liver Disease and Associated Oxidative Stress --- p.84 / Chapter 1.5 --- Objectives of the Study --- p.89 / Chapter 2. --- Materials and Methods --- p.90 / Chapter 2.1 --- Chemicals and Materials --- p.90 / Chapter 2.2 --- Animals --- p.90 / Chapter 2.3 --- Chronic Ethanol Treatment --- p.90 / Chapter 2.3.1 --- Ethanol Diet Composition --- p.90 / Chapter 2.3.2 --- Ethanol Feeding --- p.90 / Chapter 2.4 --- Monitoring of Blood Ethanol Levels --- p.96 / Chapter 2.5 --- Preparation of Microsomal Fractions --- p.96 / Chapter 2.6 --- Determination of Microsomal Protein Concentration --- p.97 / Chapter 2.7 --- Determination of Serum Aminotransferase Activities --- p.98 / Chapter 2.8 --- Liver Histology --- p.98 / Chapter 2.9 --- SDS-PAGE and Western Blot Analysis --- p.99 / Chapter 2.10 --- Hepatic Fatty Acid Composition Analysis --- p.100 / Chapter 2.10.1 --- Lipid Extraction --- p.100 / Chapter 2.10.2 --- Thin Layer Chromatography --- p.101 / Chapter 2.10.3 --- Methylation --- p.101 / Chapter 2.10.4 --- Gas Chromatography --- p.102 / Chapter 2.11 --- Statistical Analysis --- p.103 / Chapter 3. --- Results --- p.104 / Chapter 3.1 --- Average Food Consumption --- p.104 / Chapter 3.2 --- Average Ethanol Consumption for Ethanol Liquid Diet Feeding Group --- p.104 / Chapter 3.3 --- Body Weight Gain --- p.104 / Chapter 3.4 --- Blood Ethanol Levels --- p.108 / Chapter 3.5 --- "Mortality, Liver Weight and Liver Color" --- p.108 / Chapter 3.6 --- Serum ALT and AST Activities --- p.110 / Chapter 3.7 --- Liver Histology --- p.114 / Chapter 3.8 --- Western Blot Analysis --- p.119 / Chapter 3.9 --- Hepatic Lipid Fatty Acid Composition --- p.119 / Chapter 3.9.1 --- Fatty Acid Composition in Hepatic Phospholipid --- p.119 / Chapter 3.9.2 --- Fatty Acid Composition in Hepatic Triglyceride --- p.123 / Chapter 4. --- Discussion --- p.126 / Chapter 4.1 --- Nutrients Displacement after Chronic Ethanol Consumption --- p.126 / Chapter 4.2 --- Varied Blood Ethanol Levels after Chronic Ethanol Consumption --- p.127 / Chapter 4.3 --- Increase in CYP2E1 Levels after Chronic Feeding of Ethanolin WT mice --- p.127 / Chapter 4.4 --- Lack of Evidence Indicating the Development of Ethanol- Induced Liver Injury --- p.129 / Chapter 4.4.1 --- No Elevations in Serum ALT and AST Activities --- p.129 / Chapter 4.4.2 --- Normal Liver Histology --- p.130 / Chapter 4.4.3 --- Lack of Triglyceride Accumulation --- p.131 / Chapter 4.4.4 --- Elevations in Hepatic PL --- p.132 / Chapter 4.5 --- Possible Reasons for the Absence of Liver Damage after Chronic Ethanol Consumption in our Mouse Model --- p.134 / Chapter 5. --- Conclusion --- p.137 / Chapter Chapter IV --- Concluding Remarks / Chapter 1. --- A Comparison between Acute CC14 Study and Chronic Ethanol Consumption Study --- p.139 / Chapter 1.1 --- Regulation of CYP2E1 Expression --- p.139 / Chapter 1.2 --- Free Radical Production Involved in CC14- and Chronic Ethanol Consumption-Mediated Liver Injury --- p.140 / Chapter 1.3 --- An Overall Comparison between CC14 study and Chronic Ethanol Consumption Study --- p.140 / Chapter 2. --- Future Studies --- p.142 / Chapter 2.1 --- Acute CC14 Study --- p.142 / Chapter 2.1.1 --- Calcium Homeostasis Studies --- p.142 / Chapter 2.1.2 --- Spin Trapping Studies --- p.142 / Chapter 2.2 --- Chronic Ethanol Study --- p.142 / Chapter 2.2.1 --- "Generation of a Heterozygous ""Ethanol-Sensitive"" Mouse Strain (SV/129/ter x C57BL/6)" --- p.143 / Chapter 3. --- Concluding Remarks --- p.143 / References --- p.144 / Appendix --- p.167

Avasarala Sreedevi. / "December 2004." / Thesis (Ph.D.)--Chinese University of Hong Kong, 2004. / Includes bibliographical references (p. 253-272) / Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Mode of access: World Wide Web. / Abstracts in English and Chinese.

Um estudo prévio de nosso grupo de pesquisa demonstrou que uma fração eriquecida em valepotriatos obtida a partir de partes aéreas e subterrâneas de V. glechomifolia submetida à extração com CO2 supercrítico (VAL) possui efeito antidepressivo e prevenção do comportamento de doente (sickness behavior) induzido por LPS. Além disso, alguns estudos revelaram propriedades antiinflamatórias de V.wallichii e de V.amurensis. Estes dados da literatura sugerem que os valepotriatos podem ser utilizados no desenvolvimento de novos farmácos. Entretanto, dados sobre toxicidade, segurança e atividade anti-inflamatória de valepotriatos ainda são escassos. Considerando isso, o objetivo deste estudo foi investigar a atividade anti-inflamatória periférica e a toxicidade oral aguda e de doses repetidas de VAL. A atividade anti-inflamatória foi avaliada por meio do teste de formalina em camundongos CF1 e o ensaio de migração de leucócitos em ratos Wistar. Além disso, os estudos de toxicidade seguiram as normativas 423 e 407 da Organização para a Cooperação e Desenvolvimento Econômico (OECD). Diferentes grupos de camundongos foram tratados com VAL (1, 10 e 30 mg/kg), diclofenaco 50 mg/kg (controle positivo) ou saline (controle negativo) 1 h antes da injeção de formalina. No ensaio de quimiotaxia, os leucócitos foram tratados com concentrações de 0,1-1,0 μg/mL de VAL, indometacina ou diclofenaco (1 μg/mL). No estudo de toxicidade aguda, três camundongos CF1 machos foram tratados com uma dose única de VAL (2000 mg/kg, v.o.) e observados durante 14 dias. Já para o estudo de toxicidade de doses repetidas, diferentes grupos de animais (n = 10) receberam doses únicas diárias de VAL (30, 150 e 300 mg/kg, v.o.) ou veículo durante 28 dias. No teste de formalina, VAL inibiu o comportamento do tipo nociceptivo na segunda fase do teste de forma dose-dependente. O efeito da dose mais elevada de VAL foi comparável com o diclofenaco na dose de 50 mg/kg (v.o.). VAL (0,1-1 μg/mL) também inibiu a migração de leucócitos induzida por LPS (65 μg/mL) de modo dependente da concentração. Este efeito foi comparável ao efeito de indometacina (0,1 - 1 μg/mL) e superior ao efeito do diclofenaco (1 μg/mL). No estudo de toxicidade aguda apenas uma morte foi detectada, o que classifica VAL como segura (categoria 5), de acordo com a OECD-normativa 423. O estudo toxicidade de doses repetidas demonstrou que VAL na dose de 300 mg/kg retardou o ganho de peso e reduziu o consumo de ração dos animais deste grupo na primeira semana de tratamento, provavelmente devido aos efeitos sedativos da mesma. As outras doses não alteraram o ganho de peso e ingesta de ração. Nenhuma das doses de VAL alterou qualquer parâmetro comportamental, urinário, bioquímico, hematológico, anatômico ou histológico. Em conclusão, estes resultados demonstram pela primeira vez que valepotriatos, uma classe especial de terpenos que ocorrem apenas no gênero Valeriana, apresentam atividade anti-inflamatória periférica e são seguros em doses pré-clínicas eficazes, por via oral. / A previous study by our research group demonstrated that an enriched fraction obtained from the aerial and subterranean parts of V. glechomifolia submitted to supercritical CO2 extraction (VAL) shows antidepressant-like effect and prevented LPS-induced sickness behavior. Also, some studies revealed anti-inflammatory properties of V.wallichii and V.amurensis. Altogether, these findings suggest that the valepotriates scaffold might be useful to develop new antidepressant and antiinflammatory drugs. However, data about the toxicity, safety and anti-inflammatory activity of valepotriates from V. gelchomifolia are still scarce. Considering this, the aim of this study was to investigate the peripheral anti-inflammatory activity and the oral acute and repeated toxicity of VAL. The anti-inflammatory activity was assessed by using the formalin test in CF1 mice and Wistar rat’s leukocytes migration assay. Besides, the toxicity studies followed the Organization for Economic Cooperation and Development (OECD) toxicity studies guidelines 423 and 407. Different groups of mice were treated with VAL (1, 10 and 30 mg/kg), diclofenac 50 mg/kg (positive control) or saline (negative control) 1 h before the formalin injection. In the chemotaxis assay, the leukocytes were treated with a range of 0.1-1.0 μg/mL of VAL, indomethacin or diclofenac (1 μg/mL). In the acute toxicity, three CF1 mice were treated with a single dose of VAL (2000 mg/kg, p.o.) and observed for 14 days. To perform the repeated toxicity study, separated group of animals (n=10) received single daily doses of VAL (30, 150 and 300 mg/kg, p.o.) or vehicle during 28 days. In the formalin test, VAL inhibited the nociceptive behavior in the late phase in a dose dependent manner at 30mg/kg dose. The effect of the VAL highest dose was comparable to diclofenac 50 mg /kg (p.o.). VAL (0.1 - 1 μg/mL) inhibited the leukocyte migration induced by LPS (65 μg/mL) in a concentration dependent manner. This antichemotatic effect was comparable to indomethacin (0.1 – 1μg/mL) and better than diclofenac (1 μg/mL) effect. In the acute toxicity study only one death was detected, which classify VAL as safe (category 5), according to OECD-guideline 423. The repeated dose toxicity study demonstrated that VAL 300 mg/kg delayed the weight gain and reduced the food consumption in the first week, probably due to sedative effects. The other doses had no effect on weight gain and food consumption. None of doses altered any behavioral, urinary, biochemical, hematological, anatomic or histological parameters. In conclusion, these results demonstrate for the first time that valepotriates, a special class of terpenes occurring only in Valeriana genus, present peripheral anti-inflammatory activity and are safe at effective pre-clinical doses, by oral route.