La perte de l’expression de la sous-unité β4 des canaux sodiques dépendants du voltage NaV dans les tumeurs mammaires est associée à un grade cancéreux élevé et au développement des métastases. L’extinction de son expression dans les cellules MDA-MB-231 augmente de plus de deux fois leur invasivité. Au cours de cette thèse, nous avons montré que la sous-expression de β4 favorise la transition mésenchymato-amoeboïde et augmente l’invasion cancéreuse indépendante de NaV. Cette transition se caractérise par l’acquisition d’une morphologie plus arrondie, par la présence de blebs à la surface cellulaire et par une augmentation de l’activité RhoA-GTPase. Cette transition est inhibée par la surexpression du domaine intracellulaire C-terminal de la sousunité β4. L’expression de β4 peut être augmentée par un apport en acide docosahexaènoïque (22:6n-3), qui augmente l’activité du promoteur de son gène SCN4B. Le DHA augmente également l’expression de β4 en modulant l’expression des récepteurs nucléaires PPAR, sensibles aux lipides. / The loss of voltage gated sodium channel NaVβ4 subunit expression in breast cancer biopsies is associated with high grade tumors and metastatic development. The inhibition of β4 expression in MDA-MB-231 breast cancer cells enhanced their invasiveness by two fold. During this thesis, we have shown that β4 underexpression promotes mesenchymal-amoeboid transition and increases NaV-independent invasion. This transition is characterized by rounded morphology, the presence of blebs at the cell surface and an increased RhoAGTPase activity. This transition is inhibited by β4 C-terminal intracellular domain overexpression. Expression of β4 can be enhanced by a DHA supplementation that increases the encoding SCN4B promoter activity. DHA also increases β4 expression through the modulation of PPARs lipid-sensitive nuclear receptors expression.

Les récepteurs nucléaires PPAR sont régulés par les acides gras polyinsaturés n-3 (AGPI n-3), dont l’acide docosahexaènoïque (DHA, 22:6n-3) qui inhibe la croissance des lignées de cellules cancéreuses mammaires. Des siRNA et des ligands pharmacologiques des PPAR (α, β et γ) ont permis de montrer que PPARβ régule cet effet inhibiteur du DHA. Dans un modèle murin de tumeur mammaire, la supplémentation en AGPI n-3 diminue l’expression de PPARβ et les tumeurs qui régressent le plus, sous chimiothérapie, sont celles qui expriment le moins PPARβ. L’invasivité des cellules cancéreuses mammaires et l’activité du canal sodique NaV1.5, qui promeut l’invasivité, sont inhibées par le DHA. Des antagonistes de PPARβ, comme l’inhibition de son expression, diminuent l’invasivité, le courant NaV1.5 et l’activité de son partenaire, l’échangeur Na+/H+ de type 1 (NHE-1). Les activités NaV1.5 et NHE-1 sont aussi diminuées dans les cellules qui n'expriment plus PPARβ. Cette étude montrant un lien mécanistique entre DHA, PPARβ, NaV1.5 et NHE-1, indique que l’inhibition de PPARβ par des AGPI n-3 à longue chaîne pourrait compléter les traitements actuels du cancer du sein. / Peroxisome proliferator-activated receptors (PPAR) are nuclear receptors for n-3 polyunsaturated fatty acids (n-3 PUFA), such as docosahexaenoic acid (DHA, 22:6n-3) which inhibits breast cancer cell growth. Using siRNA and pharmacological ligands of PPAR (α, β or γ) in breast cancer cell lines, we showed that PPARβ regulated DHA-induced inhibition of cell growth. In a rat mammary tumor model supplemented with n-3 PUFA, PPARβ expression diminished and tumors that regressed most, under docetaxel treatment, displayed the lowest PPARβ expression. Breast cancer cells invasiveness and activity of NaV1.5 voltage-gated sodium channel, known to promote invasiveness, were both inhibited by DHA. Antagonizing PPARβ, or inhibiting its expression, reduced invasiveness and NaV1.5 current as well as its accompanying Na+/H+ exchanger type 1 (NHE-1) activity. NaV1.5 and NHE-1 activities were also reduced in cells knocked-down for PPARβ. This study shows a mechanistic link between DHA, PPARβ, NaV1.5 current and NHE-1 pro-invasive activities, advocating for the inhibition of PPARβ with long chain n-3 PUFA to complement current breast cancer therapies.

Existe um grande potencial para a expansão da produção de microalgas, tanto pela variedade de usos quanto pelos produtos que delas podem ser derivados. Dentre esses usos, as características nutricionais e funcionais conferem às algas alto valor biológico para uso em nutrição humana e animal. No entanto, são poucos os estudos que avaliam seu potencial na nutrição de peixes, mais especificadamente de tilápia, um dos principais produtos da aquicultura mundial. O objetivo deste trabalho foi avaliar o potencial de uso de duas espécies de microalgas e o nível ótimo de inclusão da Schizochytrium em dietas para a tilápia. Para tanto, foram realizados ensaios para avaliação do coeficiente de digestibilidade aparente (CDA) de uma microalga rica em proteína, Chlorella sorokiniana, e outra rica em energia, a Schizochytrium sp., ao longo de ensaios de desempenho e digestibilidade de rações contendo níveis crescentes (0%; 1%; 2,5%; 5%; 7,5% e 10%) de Schizochytrium sp. a fim de determinar o nível ótimo de inclusão nas dietas. As microalgas apresentaram altos coeficientes de digestibilidade para a tilápia, com CDA para a proteína de 90,51 e 97,20 % e para energia de 84,22 e 82,55 %, para a Chlorella e Schizochytrium, respectivamente. Os níveis crescentes de inclusão da Schizochytrium alteraram a digestibilidade das rações de forma decrescente para a proteína, passando de um CDA de 88,07 para 86,01, e energia, de um CDA de 74,19 para 67,35. No ensaio de desempenho foi registrado aumento no consumo de ração e piora na conversão alimentar aparente, sem que outros parâmetros de desempenho zootécnico fossem alterados, à medida que aumentava os níveis de inclusão da Schizochytrium na dieta. Não foram registradas alterações nos conteúdos de macro nutrientes na carcaça dos peixes, no entanto, os níveis crescentes de inclusão alteraram o perfil de ácidos graxos aumentando a quantidade de ácidos graxos n-3 no filé, principalmente do ácido docosaexaenoico (DHA), e redução na relação n-6/n-3. Dessa forma, é possível a utilizar Schizochytrium sp. como aditivo nas dietas para alteração do perfil de ácidos graxos da tilápia, enriquecendo os filés com ácidos graxos n-3, com pouco prejuízo nos parâmetros de desempenho. / There is a great potential for expand the microalgae production as by the variety of it uses as for the products and processes which may derive from them. Amongst such uses, nutritional and functional characteristics provide them a high biological value for use in human and animal nutrition. Nevertheless, few studies have evaluated their potential for fish nutrition more specifically tilapia, which is one of the main aquaculture products in the world. The aim of this research was to evaluate the potential use of two microalgae and the optimal level of inclusion of Schizochytrium in diets for tilápia. Therefore, tests were conducted to evaluate the apparent digestibility coefficient (ADC) of microalgae with high protein content Chlorella sorokiniana and another with high energy, Schizochytrium sp. and also performance assay and feed digestibility containing inclusion levels (0%, 1%, 2.5%, 5%, 7.5% and 10%) of Schizochytrium in order to determine the optimum addition level. Both species were very digestible for tilapia with ADCs for protein of 90.51 and 97.20 and for energy 84.22 and 82.55 to Chlorella and Schizochytrium respectively. The increasing levels of inclusion of Schizochytrium changed the digestibility of feed for the protein, through a ADC from 88.07 to 86.01 and for energy from 74.19 to 67.35 ADC. In the growth test as the inclusion of Schizochytrium increased there was an increase in feed intake and feed conversion ratio but any other growth parameters were changed. Regarding the carcass composition, there were no changes in macronutrients composition however, the increasing levels of Schizochytrium altered the fatty acid profile by increasing the amount of omega 3 mainly docosahexaenoic acid (DHA) and reduced the n-6/n-3 ratio. Thus, it is possible to use the Schizochytrium to modify the fatty acid profile of tilápia by incorporating omega 3, with little loss in performance parameters.

Le syndrome métabolique (MS), une association des plus dangereux facteurs de risque pour les maladies cardiovasculaires et le diabète de type 2, est devenu l'un des principaux défis cliniques et de santé publique dans le monde. Un nombre croissant d’évidences s’est accumulé au cours de la dernière décennie, démontrant l'efficacité de certains composés bioactifs alimentaires pour le traitement et la prévention du MS. Néanmoins, la plupart des études d'intervention administre les composés bioactifs sous forme de composés purs, sans considérer que l'interaction entre les bioactifs ajoutés et l'ensemble de la matrice alimentaire peut impacter la bioaccessibilité, la biodisponibilité et, par conséquent, l’efficacité de ces molécules bioactives.L'objectif principal de cette thèse, intégrée dans le projet européen PATHWAY-27, était de formuler des aliments enrichis en composés bioactifs potentiellement efficaces contre le MS, et d'étudier l'effet de la matrice alimentaire sur la bioaccessibilité et la biodisponibilité de ces bioactifs. Cette étude a mis l'accent sur l'utilisation des anthocyanes, de l’acide docosahexaénoïque et, dans une moindre mesure, des bêta-glucanes comme ingrédients bioactifs pour enrichir des produits laitiers et des produits à base d'œuf. Une combinaison des modèles de digestion in vitro et in vivo (chez le porc) a été utilisée.La composition et la structure des matrices alimentaires ont impacté la libération et la solubilisation de substances bioactives au cours de la digestion (bioaccessibilité), tel que démontré in vitro et in vivo. La structure de / Metabolic Syndrome (MS), a constellation of the most dangerous cardiovascular disease and type 2 diabetes mellitus risk factors, has become one of the major clinical and public health challenges worldwide. During the last decade, many bioactives have been proposed as effective for the treatment and prevention of MS. However, most intervention studies administer bioactives as pure compounds, without consider that bioactive-food matrix interactions could deeply impact on the bioaccessibility, bioavailability and hence on the effectiveness of bioactives.The main objective of this thesis, included within the European project PATHWAY-27, was to formulate potential effective bioactive-enriched foods against MS and to investigate the effect of the food matrix on the bioaccessibility and bioavailability of dietary bioactives.In particular, this study focused in the use of dairy and egg-based products as matrices and in the addition of anthocyanins, docosahexaenoic acid and, to a lesser extent, beta glucan as bioactives. A combination of in vitro and in vivo (pig) digestion models was used.Both the structure and the composition of the food matrices impacted the release and the solubilisation of bioactives during digestion (bioaccessibility), as demonstrated in vitro and in vivo. In addition, the structure of the food matrix modulated the final amount of DHA into the systemic circulation of pigs (bioavailability). This study proves that understanding how dietary bioactives interact with the matrix in which they are included in is the basis for the production of effective bioact

HT biomass sonicated at ES inputs < 56KJ/gTS decreased floc size by 41% and enhanced its metabolic activity by 50-250% compared to control. ES inputs >118 KJ/gTS caused HT biomass solubilization and irreversible loss of its metabolic activity and reflocculation ability. During continuous activated sludge processing (ASP) of real primary effluent the observed yield (Yobs) decreased by 20% compared to control ASP at SF (stress factor) of 1 (biomass exchanged without USPT). At SF of 0.5, 1 and 1.5 (biomass exchanged with USPT) the Yobs further decreased by 33, 25 and 44% respectively as compared to control. This indicated that combining biomass anoxic exposure with USPT enhanced sludge reduction by increasing microbial maintenance metabolism likely in combination with microbial flora shift in the ASP depending on SF.

Master of Science / Department of Biological and Agricultural Engineering / Wenqiao Yuan / Many of the current industrial fermentation processes cannot use pentose as the carbon source. However, complete substrate utilization of sugars in lignocellulose is one of the prerequisites to render economic development of biofuels or bioproducts from biomass. In this study we proposed a new process for DHA production from renewable carbon sources by first using anaerobic bacteria, Clostridium tyrobutyricum to convert pentose into organic acids with butyric acid as the main product, and then using the organic acids to feed microalgae, Crypthecodinium cohnii to produce DHA. The effect of glucose and xylose on the yield of butyric acid produced by C. tyrobutyricum was investigated, separately. Cell growth of C. tyrobutyricum increased with increasing initial glucose or xylose concentration, but was not affected significantly when the concentration was above 55g/l for glucose or 35g/l for xylose. Butyric acid yield increased as the initial sugar concentration increased in both xylose and glucose, but the conversion rate from xylose or glucose to butyric acid decreased as the sugar concentration increased. The xylose to glucose ratio in their mixture did not significantly affect cell growth or butyric acid yield. The effect of arabinose on the yield of butyric acid produced by C. tyrobutyricum was also studied. As for butyric acid production, compared with glucose or xylose, the arabinose was in a low efficiency, with butyric acid output of 2.25g/l in 10g/l arabinose and a long lag period of about 3-4 d. However, a low concentration of arabinose could be used as a nutritional supplement to improve the fermentability of a mixture of xylose and glucose. The conversion rate of sugar to butyric acid increased as the supplement arabinose increased. In order to obtain low cost xylose, corncobs were hydrolyzed and this xylose-rich product was used to culture C. tyrobutyricum. The results showed that at end of the 9 d fermentation, the concentration of butyric acid from corncob hydrolysate reached 10.56 g/l, and the mimic medium reached 11.3 g/l. This suggests that corncob hydrolysate can be used as a carbon source for butyric acid production by C. tyrobutyricum, although some inhibitory effects were found on cell growth with corncob hydrolysate. The effect of butyric acid, lactic acid and acetic acid on the yield of DHA produced by C. cohnii was also investigated, separately. The DHA yield was highly related to both biomass and DHA content in the cell, whereas lower growth rate could bring higher DHA content. The best concentration for DHA yield seemed to be 1.2g/l in three single organic acid media. In two organic acids mixture media, acetic acid tended to be beneficial for biomass accumulation, regardless whether butyric acid or lactic acid was mixed with acetic acid, the OD could reach 1.3 or above. When butyric acid was mixed with lactic acid, the highest DHA yield was achieved, due to increased DHA content from mutual influence between butyric acid and lactic acid.

Combined intakes of specific dietary fiber and fat subtypes protect against colon cancer in animal models. We evaluated associations between self-reported individual and combinations of fiber (insoluble, soluble, and pectins, specifically) and fat (omega-6, omega-3, and docosahexaenoic acid (DHA) and eicosapentaenoic acid (EPA), specifically) and colorectal cancer (CRC) risk in the Women's Health Initiative prospective cohort (n = 134,017). During a mean 11.7 years (1993-2010), 1952 incident CRC cases were identified. Cox regression models computed multivariate adjusted hazard ratios to estimate the association between dietary factors and CRC risk. Assessing fiber and fat individually, there was a modest trend for lower CRC risk with increasing intakes of total and insoluble fiber (p-trend 0.09 and 0.08). An interaction (p = 0.01) was observed between soluble fiber and DHA + EPA, with protective effects of DHA + EPA with lower intakes of soluble fiber and an attenuation at higher intakes, however this association was no longer significant after correction for multiple testing. These results suggest a modest protective effect of higher fiber intake on CRC risk, but not in combination with dietary fat subtypes. Given the robust results in preclinical models and mixed results in observational studies, controlled dietary interventions with standardized intakes are needed to better understand the interaction of specific fat and fiber subtypes on colon biology and ultimately CRC susceptibility in humans.

The objective of this research was to systematically study the effect of processing conditions on crystallization behavior and destabilization mechanisms of oil-in-water (o/w) emulsions. The effects of oil content (20 and 40 wt %); crystallization temperature (Tc = 10, 5, 0, -5, -10 °C); homogenization conditions, such as high shear (HS), very low pressure homogenization (VLPH), and high pressure homogenization (HPH); and cooling rate (0.2 and 30 °C/min) on both thermal behavior and destabilization mechanisms were analyzed. Docosahexaenoic acid (DHA) was added to VLPH emulsions and its effect on the physicochemical and oxidative stabilities and flavor was studied. Emulsions with 20% oil were less stable than those with 40% oil with a fast-cooling rate; however, stability increased when the emulsions were cooled slowly. Stability was also affected by oil and droplet size; the smaller the droplet the more stable the system. Smaller droplets (i.e., VLPH, HPH) had an effect on crystallization by delaying the onset of the crystal formation, which was promoted in emulsions with larger droplets (i.e., HS); 20% o/w emulsion crystallization was delayed more than 40%; and in emulsions crystallized using a slow-cooling rate, the crystal formation was less inhibited (i.e., crystals formed at a higher onset temperature [Ton], but at lower Tc) than when using a fast-cooling rate. The formation of lipid crystals either helped stabilize (small droplets) the emulsion and melted in a less fractionated manner or destabilized (big droplets) the emulsion. In addition, fast-cooling rates have greater fractionation than slow-cooling rates. Due to the greater stability of VLPH emulsions after thawing from being at -10 °C for 3 h, DHA was added to evaluate its effect on flavor (besides the effect on stability) of the emulsion. A descriptive panel was used to evaluate four attributes: oxidized, rancid, fishy, and buttery. The panelists were given samples after 72 h, because contrary to the TBA analysis which showed no significant differences between samples with and without DHA, the fishy smell was evident. The sensory evaluation results showed that there was a significant (p < 0.05) difference in fishiness between the VLPH emulsions with and without DHA, and that the odor was repulsive. No significance was seen for rancid and buttery flavors, and only a marginal significance was seen for oxidized.

HT biomass sonicated at ES inputs < 56KJ/gTS decreased floc size by 41% and enhanced its metabolic activity by 50-250% compared to control. ES inputs >118 KJ/gTS caused HT biomass solubilization and irreversible loss of its metabolic activity and reflocculation ability. During continuous activated sludge processing (ASP) of real primary effluent the observed yield (Yobs) decreased by 20% compared to control ASP at SF (stress factor) of 1 (biomass exchanged without USPT). At SF of 0.5, 1 and 1.5 (biomass exchanged with USPT) the Yobs further decreased by 33, 25 and 44% respectively as compared to control. This indicated that combining biomass anoxic exposure with USPT enhanced sludge reduction by increasing microbial maintenance metabolism likely in combination with microbial flora shift in the ASP depending on SF.

The causative agent of malignant tertian malaria, Plasmodium falciparum undergoes an arrested growth phenotype of its erythrocytic stage when under drug-stress. Recent artemisinin treatment failures seem to be indicative of such induction followed by recrudescence rather than actual therapeutic failure. Likewise, P. vivax hypnozoites are the prototypic dormants and the latent infections for which they are responsible prove most difficult to treat. Dihydroartemisinin, an artemisinin-derivative, can be used to exploit this mechanism by inducing a dormant state in ring-stage P. falciparum parasites and in turn, their recovery may be used as a screening period for compounds that inhibit or foster growth. Specifically, parasites stably transfected with luciferase were used to quantitatively observe growth (or lack thereof) response of parasites to the phytohormone gibberellic acid and the herbicide, fluridone. Using their behavior as comparative controls, the Medicines for Malaria Venture (MMV) Malaria Box was screened for similar activity. The most active compound, 1,2,3,4-tetrahydroacridin-9-ol a quinoline-derivate caused cells to wake even earlier than expected. Since quinine and other such drugs have historically been most effective in treating malaria, it seems appropriate that such a finding was made. Following this the MMV Box was screened again against uninduced 3D7 parasites to determine if any were capable of causing a dormant response under the hypothesis that such a reaction is a defensive adaptation of P. falciparum. Four compounds were found to be active of which two appear to be inducing dormancy in the second cycle rather than the first akin to DHA. These quiescent periods also appear to be shorter indicating that the latter is more efficient. It is possible that given the length of interaction with artemisinin, P. falciparum is more adept to respond to its derivatives likewise the mechanism of action may be different enough to change the nature of the response.