The seed of Santalum spicatum is rich in a fixed oil (59% by weight), which is characterised by a high percentage of acetylenic, ethylenic ximenynic acid (35% of total fatty acids). A number of important aspects of the seed fixed oil, its composition in developing seeds, its triacylglycerols molecular species in the oil, the nutrition and toxicity of the oil feeding, and the possible bioactivity of ximenynic acid in mice were investigated.The identification of cis and trans isomers of ximenynic acid in the seed oil, and the metabolite of ximenynic acid in mouse liver lipid fractions were achieved using 2-amino-2-methyl-1-propanol to form 2-substituted 4,4-dimethyloxazoline derivatives, which were analysed by gas chromatography with mass spectrometric detection.Changes in proximate and fatty acid composition were investigated in developing seed collected weekly from about seven days after flowering to maturity. It was determined that moisture and carbohydrate contents decreased significantly during the development sequence, while fixed oil content increased from 0.3% to 50% (by weight) with seed development. A corresponding increase in the proportions of both oleic and ximenynic acids occurred suggesting a precursor/product relationship. Mature seed collected from different locations in Western Australia showed minor differences in characteristics and lipid composition, which may have been influenced by geographical origin and harvesting year of samples.The lipid components from the seed oil were separated using thin-layer chromatography and the individual triglyceride bands were characterised by high performance liquid chromatography and gas chromatography using flame ionisation and mass spectrometric detection after removal from the plate. The triximenynin (trisantalbin) band showed no other contaminating fatty acids and was obtained in a relatively pure state.A ++ / nutrition and toxicity study was performed by feeding a semi-synthetic diet containing sandalwood seed oil to a level of 15% of total energy content to a group of mice for one month and another group for two months. The most significant effect of sandalwood seed oil ingestion when compared with a standard lab diet (5% fat, by weight) and a canola oil-enriched diet (15% fat, by weight) was an apparent reduction in body weight gain, which may be the effect of ximenynic acid as a growth retardant. Serum aspartate aminotransferase levels were determined in the mice as an indicator of hepatotoxicity. These levels were higher in mice fed the sandalwood seed oil diet than those fed the standard lab diet, suggesting that ximenynic acid may affect liver-specific enzyme activity. Analysis of the total lipid fatty acids of various tissues and organs of mice showed only a low incorporation of ximenynic acid into the general tissues (0.3-3% by weight), and its absence in the brain.This study suggests a few health benefits from consumption of large quantities of sandalwood seed oil in the diet. These include a low lipid content in blood, heart, muscle, increase in the 16:1/16:0 and 18:1/18:0 ratios, production of increased levels of 18:1 (n-9) and docosahexaenoic acid, and decreased levels of arachidonic acid in certain tissues. There were no specific pathological, morphological or mortality changes observed in the mice.Sandalwood seed may be both a food and a medicine.

O presente estudo visou avaliar os efeitos da administração do resíduo aquoso final (RAF) da Ipomoea carnea e da suainsonina, sobre o sistema imune de ratos jovens em idade pré-pubere (21 dias) e adultos (70 dias). Para isso, o RAF foi administrado nas doses de 1,0; 3,0 e 7,0 g/kg e a suainsonina na dose de 5,0 mg/kg, por gavage, durante 14 dias para avaliação histopatológica de diferentes órgãos, peso relativo de baço e timo, celularidade de medula óssea, fenotipagem de linfócitos T e B presentes no sangue, timo e baço, e atividade proliferativa de linfócitos T de animais tratados apenas com suainsonina. Além disto, foi realizada toxicocinética da suainsonina de animais jovens e adultos. Dentre os resultados obtidos com o tratamento com RAF, foi verificado que todos os animais apresentaram lesões, tais como: congestão hepática e esplênica e vacuolização renal. Em ratos adultos observou-se ainda diminuição do ganho de peso e do consumo de ração, além de involução tímica e diminuição da celularidade da medula óssea. Já nos jovens, as principais alterações observadas foram quanto à alteração fenotípica das populações de linfócitos T CD8+ no timo, sangue e baço e de linfócitos B no sangue e baço. Entretanto, estas alterações não podem ser atribuídas exclusivamente ao RAF de I. carnea, pois os animais do grupo pear-feeding (grupo sob restrição alimentar para mimetizar a diminuição da ingesta de alimentos daqueles ratos tratados com a maior dose do RAF) apresentaram resultados semelhantes àqueles tratados com a planta. Em relação aos resultados obtidos em ratos jovens tratados com suainsonina, verificou-se menor porcentagem de linfócitos B no baço e no sangue e maior expansão clonal de linfócitos T CD4+ de ratos imunizados in vivo e desafiados ex vivo com anatoxina tetânica. Por fim, o estudo toxicocinético revelou que este alcalóide possui biodisponibilidade de apenas 9 horas tanto nos animais jovens quanto nos adultos. Assim, é possível concluir que a suainsonina, principal princípio ativo da I. carnea, tem atividade imunomodulatória, mas esta atividade difere do efeito imunomodulador causado pelo RAF provavelmente devido a presença de outros alcalóides também encontrados na planta / This study aimed to evaluate the effects of administration of the final aqueous residue (RAF) of Ipomoea carnea and swainsonine on the immune system of young (21 days) and adult rats (70 days). For this, the RAF was administered at doses of 1.0, 3.0 and 7.0 g/kg and swainsonine at dose of 5.0 mg/kg by gavage for 14 days for histopathological evaluation of different organs, relative weight of spleen and thymus, bone marrow cellularity, phenotyping of T and B lymphocytes in the blood, thymus and spleen, and proliferative activity of T lymphocytes from animals treated with swainsonine. Moreover, we performed toxicokinetics of swainsonine in young and adult rats. Among the results obtained with treatment with the RAF, it was observed that all animals had lesions such as hepatic and splenic congestion and kidney vacuolization. In adult rats there was also reduction in weight gain and feed intake, thymic involution and decreased bone marrow cellularity. Already in the young rats, the main changes observed were in the populations of T CD8+ lymphocytes in the thymus, spleen and blood and B lymphocytes in the blood and spleen. However, these changes cannot be attributed solely to the RAF of I. carnea, because the animals of the pear-feeding group (group under food restriction to mimic the decrease in food intake of those rats treated with the higher dose of the RAF) had similar results to those treated with the plant. Regarding the results obtained in young rats treated with swainsonine, there was a lower percentage of B lymphocytes in the spleen and blood and increased clonal expansion of T CD4+ lymphocytes from rats immunized in vivo and challenged ex vivo with tetanus toxoid. Finally, the toxicokinetic study has revealed that bioavailability of this alkaloid is only 9 hours in the both animals young and adult. Thus, we conclude that swainsonine, the main active principle of I. carnea, has immunomodulatory activity, but this activity differs from the immunomodulatory effect caused by the RAF probably due to the presence of other alkaloids also found in the plant

O presente estudo visou avaliar os efeitos da administração do resíduo aquoso final (RAF) da Ipomoea carnea e da suainsonina, sobre o sistema imune de ratos jovens em idade pré-pubere (21 dias) e adultos (70 dias). Para isso, o RAF foi administrado nas doses de 1,0; 3,0 e 7,0 g/kg e a suainsonina na dose de 5,0 mg/kg, por gavage, durante 14 dias para avaliação histopatológica de diferentes órgãos, peso relativo de baço e timo, celularidade de medula óssea, fenotipagem de linfócitos T e B presentes no sangue, timo e baço, e atividade proliferativa de linfócitos T de animais tratados apenas com suainsonina. Além disto, foi realizada toxicocinética da suainsonina de animais jovens e adultos. Dentre os resultados obtidos com o tratamento com RAF, foi verificado que todos os animais apresentaram lesões, tais como: congestão hepática e esplênica e vacuolização renal. Em ratos adultos observou-se ainda diminuição do ganho de peso e do consumo de ração, além de involução tímica e diminuição da celularidade da medula óssea. Já nos jovens, as principais alterações observadas foram quanto à alteração fenotípica das populações de linfócitos T CD8+ no timo, sangue e baço e de linfócitos B no sangue e baço. Entretanto, estas alterações não podem ser atribuídas exclusivamente ao RAF de I. carnea, pois os animais do grupo pear-feeding (grupo sob restrição alimentar para mimetizar a diminuição da ingesta de alimentos daqueles ratos tratados com a maior dose do RAF) apresentaram resultados semelhantes àqueles tratados com a planta. Em relação aos resultados obtidos em ratos jovens tratados com suainsonina, verificou-se menor porcentagem de linfócitos B no baço e no sangue e maior expansão clonal de linfócitos T CD4+ de ratos imunizados in vivo e desafiados ex vivo com anatoxina tetânica. Por fim, o estudo toxicocinético revelou que este alcalóide possui biodisponibilidade de apenas 9 horas tanto nos animais jovens quanto nos adultos. Assim, é possível concluir que a suainsonina, principal princípio ativo da I. carnea, tem atividade imunomodulatória, mas esta atividade difere do efeito imunomodulador causado pelo RAF provavelmente devido a presença de outros alcalóides também encontrados na planta / This study aimed to evaluate the effects of administration of the final aqueous residue (RAF) of Ipomoea carnea and swainsonine on the immune system of young (21 days) and adult rats (70 days). For this, the RAF was administered at doses of 1.0, 3.0 and 7.0 g/kg and swainsonine at dose of 5.0 mg/kg by gavage for 14 days for histopathological evaluation of different organs, relative weight of spleen and thymus, bone marrow cellularity, phenotyping of T and B lymphocytes in the blood, thymus and spleen, and proliferative activity of T lymphocytes from animals treated with swainsonine. Moreover, we performed toxicokinetics of swainsonine in young and adult rats. Among the results obtained with treatment with the RAF, it was observed that all animals had lesions such as hepatic and splenic congestion and kidney vacuolization. In adult rats there was also reduction in weight gain and feed intake, thymic involution and decreased bone marrow cellularity. Already in the young rats, the main changes observed were in the populations of T CD8+ lymphocytes in the thymus, spleen and blood and B lymphocytes in the blood and spleen. However, these changes cannot be attributed solely to the RAF of I. carnea, because the animals of the pear-feeding group (group under food restriction to mimic the decrease in food intake of those rats treated with the higher dose of the RAF) had similar results to those treated with the plant. Regarding the results obtained in young rats treated with swainsonine, there was a lower percentage of B lymphocytes in the spleen and blood and increased clonal expansion of T CD4+ lymphocytes from rats immunized in vivo and challenged ex vivo with tetanus toxoid. Finally, the toxicokinetic study has revealed that bioavailability of this alkaloid is only 9 hours in the both animals young and adult. Thus, we conclude that swainsonine, the main active principle of I. carnea, has immunomodulatory activity, but this activity differs from the immunomodulatory effect caused by the RAF probably due to the presence of other alkaloids also found in the plant

De nombreuses substances chimiques sont utilisées par l’industrie cosmétique pour entrer dans la composition de formules. En dehors de la nécessité d’évaluer leur efficacité, l’industrie cosmétique se doit surtout d’évaluer la sécurité de leurs substances pour l’humain. L'évaluation toxicologique des substances chimiques est réalisée dans le but de révéler un effet toxique potentiel de la substance testée. Parmi les effets potentiels que l’on souhaite détecter, la toxicité du développement (tératogénicité), c’est-à-dire la capacité d’une substance à provoquer l’apparition d’anomalies lors du développement embryonnaire, est fondamentale. En accord avec les législations internationales qui interdisent à l’industrie cosmétique d’avoir recours à des tests sur animaux de laboratoire pour l’évaluation de leurs substances, l’évaluation toxicologique de ces substances se base sur les résultats de tests in silico, in vitro et de tests faits sur des modèles alternatifs aux animaux de laboratoire. Pour le moment cependant, peu de méthodes alternatives existent et ont été validées pour la toxicologie du développement. Le développement de nouvelles méthodes alternatives est donc requis. D'autre part, en plus de l’évaluation de la sécurité des substances chez l’humain, l’évaluation de la toxicité pour l’environnement est nécessaire. L’usage de la plupart des produits cosmétiques et d’hygiène corporelle conduit, après lavage et rinçage, à un rejet à l’égout et donc dans les cours d’eau. Il en résulte que les environnements aquatiques (eaux de surface et milieux marins côtiers) sont parfois exposés aux substances chimiques incluses dans les formules cosmétiques. Ainsi, l’évaluation toxicologique environnementale des cosmétiques et de leurs ingrédients nécessite de connaître leur toxicité sur des organismes représentatifs de chaînes alimentaires aquatiques. Dans ce contexte, le modèle embryon de poisson présente un double avantage pour l’industrie cosmétique. Ce modèle, jugé par les législations internationales comme étant éthiquement acceptable pour les évaluations toxicologiques réalisées par l’industrie cosmétique, est représentatif des organismes aquatiques. Il est donc pertinent pour évaluer la toxicité environnementale des substances chimiques. D'autre part, ce modèle apparaît prometteur pour évaluer l’effet tératogène de substances chimiques chez l’humain. Pour ces raisons, un test d’analyse de la tératogénicité des substances chimiques est actuellement développé. Ce test se base sur l’analyse d’embryons de medaka (Oryzias Latipes) à 9 jours post fertilisation, après exposition des embryons par balnéation à des substances à concentrations déterminées. L’analyse de paramètres fonctionnels et morphologiques conduit au calcul d’un indice tératogène, qui permet de tirer une conclusion quant à l’effet tératogène de la substance testée. Cet indice est calculé à partir des mesures du taux de mortalité et du taux de malformations chez les embryons. L’objectif de ce projet est d’automatiser le test d’analyse de la tératogénicité, par classification automatique des embryons faite à partir d’image et de vidéo. La première méthode développée concerne la détection des battements cardiaques à partir de séquences vidéos, dans le but de calculer le taux de mortalité. Nous nous sommes ensuite concentrés sur deux types de malformations courantes qui sont les malformations axiales, et l'absence de vessie natatoire, en utilisant une méthode d'apprentissage automatique. Cette analyse doit être complétée par l'analyse d'autres malformations et conduire à un calcul du taux de malformations et de l’indice tératogène pour la substance testée / Numerous chemicals are used as ingredients by the cosmetics industry and are included in cosmetics formula. Aside from the assessment of their efficacy, the cosmetics industry especially needs to assess the safety of their chemicals for human. Toxicological screening of chemicals is performed with the aim of revealing the potential toxic effect of the tested chemical. Among the potential effects we want to detect, the developmental toxicity of the chemical (teratogenicity), meaning its capability of provoking abnormalities during the embryonic development, is crucial. With respect to the international regulations that forbid the use of animal testing for the safety assessment of cosmetics, the toxicological assessment of chemicals must base on an ensemble of in silico assays, in vitro assays and alternative models based assays. For now, a few alternative methods have been validated in the field of developmental toxicology. The development of new alternative methods is thus required. In addition to the safety assessment, the environmental toxicity assessment is also required. The use of most of cosmetics and personal care products leads to their rejection in waterways after washing and rince. This results in the exposition of some aquatic environments (surface waters and coastal marine environments) to chemicals included in cosmetics and personal care products. Thus, the environmental assessment of cosmetics and of their ingredients requires the knowledge of their toxicity on organisms that are representative of aquatic food chains. In this context, the fish embryo model, which is ethically acceptable according to international regulations, presents a dual advantage for the cosmetics industry. Firstly, as a model representative of aquatic organisms, it is accurate for the environmental assessment of chemicals. Secondly, this model is promising for the assessment of the teratogenic effect of chemicals on human. For this reason, a teratogenicity assessment test is developed. This test is based on the analysis of medaka fish embryos (Oryzias Latipes) at 9 days post fertilization, after balneation in a predetermined concentration of the chemical under study. The analysis of functional and morphological parameters allows to calculate a teratogenicity index, that depends on both rates of dead and malformed embryos. This index allows to to draw a conclusion concerning the teratogenic effect of the chemical.The objective of this project is to automate the teratogenicity test, by automated image and video classification. A first method is developed that aims to automatically detect embryo heart beats from acquired video sequences. This method will allow to calculate the proportion of dead embryos. We then focus on the detection of two common malformations: axial malformations and absence of a swim bladder, based on a machine learning classification. This analysis must be completed by the detection of other malformations so that we can measure the rate of malformed embryos and thus, calculate the teratogenicity index of the tested chemical