On étudie les potentialités d'action de la consommation de jus de fruits et de vins riches en polyphénols sur modèle pathologique de stress oxydant, l'athérosclérose précoce, d'origine nutritionnelle, ainsi que leur effet protecteur spécifique vis à vis de cette pathologie. Dans une première partie expérimentale, nous avons analysé les polyphénols de trois variétés de jus de framboises (Cardinal, Glen Ample et Tulameen) et deux vins (Kaki et Merlot) par HPLC. Nous avons identifié et quantifié les composants phénoliques de la famille des anthocyanes et des ellagitanins, les deux constituants majeurs des polyphénols de framboise. Leurs compositions diffèrent significativement, en particulier entre Glen Ample et Tulameen. Concernant les vins, les flavan-3-ols (monomères et oligomères) ont été analysés. Les concentrations de chaque composant étaient fondamentalement différents entre les deux vins, sauf pour les dimères. Ceci nous a conduit à aborder l'étude in vivo sur hamster syrien doré recevant un régime riche en lipides et cholestérol, et carencé en antioxydants alimentaires. L'ingestion de Glen Ample (équivalent à 250 mL de jus de framboise par jour pour une homme de 70 kg) inhibe les facteurs pro-oxydants et améliore le statut antioxydant alors que le jus Tulameen a un effet protecteur contre la dyslipidémie. Ces effets peuvent être liés aux ellagitanins et anthocyanes respectivement. Nous avons montré que la consommation de vin à dose nutritionnelle (équivalent à 2 verres par repas pour un homme de 70 kg) améliorait la réactivité vasculaire, la dyslipidémie et le statut antioxydant. Parmi ces effets bénéfiques, aucune différence n'apparaît entre le vin de kaki et le Merlot, ce qui suggère que les dimères de procyanidines sont impliqués dans les effets biologiques des polyphénols. / The purpose of this study is to explore the potential of fruit juices and wines consumption rich in polyphenol on pathological models of oxidative stress, diet-induced early atherosclerosis, and to research their specific protective effect against the pathology.In the first part of this work we analyzed the polyphenols in three varieties of raspberry juices (Cardinal, Glen Ample and Tulameen) and two wines (Persimmon and Merlot) by HPLC. We identified and quantified phenolic components of anthocyanins and ellagitannins family, the two major constituents in raspberries polyphenols. Their compositions differed significantly especially between Glen Ample and Tulameen. For wines, flavan-3-ols (monomers and oligomers) were analyzed. Concentrations of each component were fundamentally different in two wines except for procyanidin dimers.These results led us to address in vivo studies on Syrian Golden hamsters fed a high-fat, high-cholesterol diet deprived in dietary antioxidants. Consumption of Glen Ample (equivalent to 250 mL of raspberry juice per day for a 70 kg human) inhibits pro-oxidant factors and improves antioxidant status whereas Tulameen shows a protective effect against dyslipidemia which may be due to its ellagitannins and anthocyanins respectively.Nutritional dose of wines (equivalent to 2 glasses of wine per meal for a 70 kg human) improves vascular reactivity, dyslipidemia and antioxidant status. No difference appeared between persimmon and merlot wine antioxidant properties suggesting that procyanidin dimers are implicated in the biological effects of polyphenols.

Various studies suggest tannins act as antioxidants, anticarcinogens, cardio-protectants, anti-inflammatory agents, and antimicrobials. However, more investigation is needed to examine the bioavailability of tannins. Tannins bind to salivary peptides by hydrophilic and hydrophobic mechanisms. Electrospray Ionization Mass Spectrometry (ESI-MS) has been used to assess both hydrophilic and hydrophobic components of protein complexes. ESI-MS could potentially be an effective tool for screening the bioavailability of tannins. Weaker binding tannins are predicted to be more highly absorbed by the body, and should therefore exhibit greater bioavailability. Rannulu and Cole have used ESI-MS to measure binding affinities of procyanidin tannin stereoisomers for salivary peptides in aqueous solution. The condensed tannins procyanidin B1, B2, B3, and B4 demonstrated significantly different binding affinities (binding strengths) for the Histatin 5 salivary peptide. The procyanidin-Histatin 5 binding mechanisms in the ESI-MS experiments by Rannulu and Cole were investigated using the FRED docking program combined with molecular dynamics optimization in the AMBER software suite. The simulations suggest residual liquid-phase binding interactions in procyanidin-Histatin 5 complexes are maintained in the gas phase under conditions resembling those in ESI-MS experiments, though the gas-phase interaction energies were enhanced. Increased hydrogen bonding and decreased π-π stacking interactions were also detected in gas versus liquid-phase procyanidin-Histatin 5 complexes. In addition, simulation results suggest multiple conformations of procyanidins bind Histatin 5 at several sites and procyanidin binding does not fix the Histatin 5 peptide backbone. The simulations agree with previous studies which indicate aromatic Histatin 5 residues are responsible for procyanidin-Histatin 5 binding and tannins can bind salivary peptides in multiple conformations. The effects of Hofmeister salts on complexation of an amphiphilic guest adamantane carboxylic acid to the hydrophobic surface of a deep-cavity cavitand have been investigated by Gibb et al. Adamantane-cavitand binding was found to be largely enthalpically driven, though adamantane binding in the presence of the salting-in anions perchlorate and thiocyanate was entropically driven. Gibb et al. also found that perchlorate-cavitand binding was enthalpically favorable, though entropically unfavorable. Potential-of-mean-force (PMF) calculations for perchlorate-cavitand and thiocyanate-cavitand complexation were performed using umbrella sampling with a modified version of the sander module from the Amber 9 software suite to further investigate the thermodynamic properties of Hofmeister-anion binding to the hydrophobic cavitand. The enthalpy for salting-in anion-cavitand complexation was calculated from the potential energy difference between the bound and unbound state (the potential energy of binding) along with the entropy. The binding entropy and enthalpy were also calculated using a finite difference approximation to the entropy. The enthalpy for perchlorate-cavitand complexation calculated from the binding energy and the finite difference approximation to the entropy was favorable with an unfavorable entropy. The binding enthalpy and entropy for thiocyanate-cavitand complexation calculated from the binding energy and finite difference approximation to the entropy were unfavorable and favorable, respectively, perhaps due to a classical hydrophobic effect. The orientation of the ligand, the number of water molecules displaced from the ligand and cavitand upon complexation, and the number of nearest-neighbor atom contacts between the ligand and the cavitand were also calculated. Additionally, the energetics of various interactions involved in salting-in anion-cavitand complexation including the anion-cavitand, anion-water, cavitand-water, and water-water interactions were assessed, though the data were inconclusive.

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Dans un contexte de réduction des intrants chimiques, l’utilisation des phytosanitaires naturels stimulant l’immunité végétale ouvre la porte vers une nouvelle approche de protection des plantes. Ces composés éliciteurs regroupés sous le terme de «Stimulateurs des Défenses naturelles des Plantes» (SDP) activent le système défensif de la plante la rendant plus résistante aux bio-agresseurs. Les SDP, de nature diverse, se présentent sous forme de composés uniques ou en mélange dans les extraits végétaux. Au cours de ma thèse, nous avons démontré l’activité SDP des extraits de marc de raisin. Les extraits issus de sous-produits de la vigne, marc de raisin rouge, marc de raisin blanc et pépins de raisin induisent diverses réactions de défense au sein de plantes modèles. Nous avons focalisé notre étude sur l’extrait de marc de raisin rouge (EMR) stimulant l’immunité chez le tabac. Infiltré sur feuilles, l’EMR induit la réponse de type HR caractérisée par l’apparition de lésions chlorotiques et accumulation de composés autofluorescents dans les tissus infiltrés. Ces réactions de défense locales ont été observées également chez l’arabette et la tomate. L’EMR déclenche les réponses LAR et SAR avec l’accumulation des transcrits des gènes de défense dans les feuilles de tabac et ce quelque soit son mode d’application (infiltration ou pulvérisation). Le mode d’action de l’EMR a été abordé sur cultures cellulaires de tabac BY-2. L’EMR induit une forte alcalinisation du milieu extracellulaire avec une mobilisation du calcium (Ca2+), l’expression des gènes de défense et la mort cellulaire. Une étude pharmacologique de la mort cellulaire suggère la mise en place de mort cellulaire programmée (PCD) dans les cellules de tabac. La caractérisation de la voie de signalisation activée par l’EMR a été étudiée avec le mutant NahG de tabac incapable d’accumuler l’acide salicylique (SA). Les réponses de défense (HR, LAR et SAR) sont faiblement induites par l’EMR chez le mutant nahG. L’EMR provoque une réponse de type HR fortement réduite avec une faible accumulation des composés autofluorescents et une diminution drastique de l’accumulation des transcrits des gènes PR suggérant l’intervention du SA dans l’induction des réactions de défense. Le degré de protection induit par l’EMR a été déterminé sur le pathosystème tabac/Phytophthora parasitica. Pulvérisé sur feuilles, l’EMR réduit de 45% les zones infectées par l’oomycète. Ce degré de protection semble être le résultat de l’activité antimicrobienne de l’EMR combinée à l’activité SDP. Aucune protection n’a été observée chez le mutant nahG confirmant l’implication de SA dans la résistance induite par l’EMR. Le fractionnement de l’EMR a permis de simplifier la formule active des extraits de raisin et d’identifier un mélange de molécules potentiellement capables d’induire l’activité SDP. Les composés actifs sont de nature polyphénolique et contiennent de la procyanidine B2 capable à elle seule d’induire la réponse de type HR et l’expression de l’antimicrobien PR1. Cependant, il semble que cette molécule agisse en association avec d’autres composés polyphénoliques pour stimuler le système défensif de la plante. / In order to reduce chemical inputs, the use of natural phytosanitary products stimulating plant immunity are emerging approaches in phytoprotection. These elicitor compounds known as "Plant Defense Inducers" (PDI) activate the plant defense system and improve their resistance to pests attack. PDI are single molecule or mixture of compounds extracted from plant. In my thesis, we demonstrated the PDI activity of different grape marc extracts. The winery byproducts, red grape marc extract, white grape marc extract and grape seed extract all induced various defense reactions in several plant models. We focused our study on the red grape marc extract (GME) which stimulates the immunity system in tobacco plants. When infiltrated into tobacco leaves, GME induced HR-like response characterized by the appearance of chlorotic lesions and accumulation of autofluorescent compounds in infiltrated tissues. Similar local defense reactions have been observed in Arabidopsis thaliana and tomato. GME also triggered LAR and SAR responses and induced defense gene transcript accumulation in tobacco leaves after infiltration or spraying. The GME mode of action was studied using the suspension-cultured cells of tobacco BY-2. GME induced rapid alkalinization of extracellular medium with calcium mobilization, expression of defense genes and cell death. A pharmacological approach of this defensive phenomenon suggests the establishment of programmed cell death (PCD) in tobacco cells. The characterization of the signaling pathway activated by GME was studied using tobacco nahG mutant unable to accumulate salicylic acid (SA). Defense responses (HR, LAR and SAR) induced by GME were impaired in the nahG mutant. GME drastically reduced HR-like response symptoms and PR transcript accumulation. These data suggest the implication of SA in the GME-induced plant defense reactions. The GME-induced protection was evaluated in the model pathosystem of compatible interaction between Nicotiana tabacum and Phytophthora parasitica var. nicotianae (Ppn). GME could reduce by 45% the infected areas induced by the oomycete on tobacco leaves. This level of protection was the result of the combined antimicrobial and PDI actions of GME. GME had no protecting effect against Ppn on NahG leaves evidencing the involvement of SA in the GME-induced resistance. GME fractionation led to identification of a bioactive molecule mixture capable of inducing the PDI activity. The active compounds are polyphenolics and involve procyanidin B2 which is by itself able to induce the HR-like response and PR1 transcript accumulation. This compound should act in combination with other polyphenolic molecules to stimulate the full plant defense reactions.

Les polyphénols sont des métabolites secondaires d’origine végétale. Ces substances naturelles connues pour leurs pouvoirs antioxydants et anti-radicalaires, contribuent à la protection de la santé humaine notamment contre les maladies cardiovasculaires et neurodégénératives, mais également contre certains cancers et diabètes. Dans certains cas, ces effets biologiques bénéfiques pour la santé pourraient également être liés à une interaction spécifique polyphénol-protéine peu étudiée à ce jour par manque d’outils moléculaires adaptés. Les travaux effectués au cours de cette thèse ont consisté à concevoir, à synthétiser et à évaluer des sondes moléculaires polyvalentes porteuses de polyphénols comme substrats d’affinité pour l’analyse des interactions polyphénol-protéine. Dans ce contexte, de nombreuses sondes arborant différents types de polyphénols ont été synthétisées. Ces différentes sondes pourront être utilisées en protéomique chimique du type "Affinity-Based Protein Profiling" (ABPP) pour identifier au sein d’un mélange complexe de protéines, une protéine ayant une affinité spécifique pour un polyphénol donné. Ces mêmes sondes permettront également d’étudier de manière qualitative les interactions d’un polyphénol avec une protéine donnée en temps réel par la technique de résonance plasmonique de surface (SPR). / Polyphenols are plant secondary metabolites. These natural substances, known for their antioxidant and anti-free radical properties, generally contribute to the protection of human health not only against cardiovascular and neurodegenerative diseases, but also against certain cancers and diabetes. In some cases, these beneficial biological effects could also be related to specific polyphenol-protein interactions. However, studying this type of interactions has suffered from the lack of adequate molecular tools. The work carried out during this thesis has included designing, synthesizing and evaluating modulable polyfunctional molecular probes carrying polyphenols as affinity substrates to analyze polyphenol-protein interactions. In this context, various probes harboring different kinds of polyphenols were synthesized. These probes could be used in chemical proteomics following an “Affinity-Based Protein Profiling” approach (ABPP) to identify a protein within complex protein mixtures, which has a specific affinity with a given polyphenol. These probes will also allow studying the interactions of a polyphenol with a given protein in real time in a qualitative way by surface plasmon resonance (SPR).

Les mousses rigides constituent une classe très importante de matériaux de par leur large champ d’applications et de leur poids économique considérable. Le développement de mousses à base de tannins a permis d’obtenir des matériaux issus à 90% de produits naturels. Dans ce travail, un type de tannins très réactif dit « procyanidine », a été employé. Les formulations et la technologie de fabrication de mousses rigides ont été développées à partir de tannins extraits d’écorce de pin et d’épicéa. Dans un premier temps, la composition et la réactivité des tannins ont été étudiées. Ensuite, l’étude du processus de moussage des résines à base de ces tannins a permis l’élaboration d’une large gamme de mousses rigides tannin-furanique. Leur excellente résistance au feu, leur résistance mécanique et leur haute performance thermique en font des matériaux d’isolation de haute qualité. Ces mousses peuvent également être employées comme absorbeurs acoustiques. De plus, de nouveaux solides cellulaires à base de tannins et de protéine ont également été développés, proposant ainsi un nouveau type de matériaux issu de produits naturels / Rigid foams contitute a very important class of materials considering their wide application range and their considerable economic impact. The development of tannin-based foams provided materials made from 90% natural products.In this work, a type of tannins highly reactive, namely « procyanidin », was used. First, tannins composition and reactivity were studied. Then, the foaming process investigation of tannin-based resins leads to a wide pannel of tannin-furanic rigid foams. Their excellent fire resistance, mechanical resistance and high thermal performance make them high quality insulation materials. These foams are also suitable for other applications such acoustic absorption. Thus, new cellular solids from tannin and protein were also developped, offering a new type of materials derived from natural products

Consumption of cocoa and dark chocolate products has been associated with positive health outcomes including reduced onset of cardiovascular disease, inflammation, diabetes, obesity, and platelet disorders. Cocoa polyphenols, putatively responsible for these beneficial activities, are highly impacted by cocoa variety, agronomic effects and processing history. However, the difference in polyphenol concentration and composition between cocoa products originating from different hybrid clones (selected for high yield) or from different fermentation conditions is not fully understood. Detailed polyphenol characterization including determination of total polyphenol and total procyanidin concentrations, and qualitative and quantitative analysis of (mean) degree of polymerization was conducted. Significant differences in total polyphenol and procyanidin concentrations were observed between five genetic clones grown by the USDA-ARS Cocoa Germplasm Repository located in Mayagüez, Puerto Rico. To facilitate cocoa fermentation research in laboratories distant from cocoa harvesting sites, a laboratory-scale cocoa fermentation model system was developed in this study. This model system used dried, unfermented, cocoa beans and simulated pulp medium as the starting material. The model system supported growth of the essential succession of cocoa fermenting microorganisms and generated similar chemical changes to those observed in on-farm cocoa fermentation. Using this model system, the impact of inoculation with proprietary yeast strains Saccharomyces cerevisiae Lev F and Saccharomyces cerevisiae Lev B on cocoa polyphenol concentration and composition was evaluated. Inoculation with both yeast strains resulted in increased fermentation rate and Lev B inoculation resulted in higher total polyphenol and procyandin contents at the end of fermentation. Overall, the present work addressed the influence of cocoa variety selection and fermentation process conditions on the composition and concentration of polyphenols. These findings will contribute to continued efforts to develop cocoa products with optimized bioactivity and maximum disease preventative effects. / PHD / Annual worldwide cocoa production exceeds four million tons and the cocoa market impacts not only cocoa growing regions, but also countries producing cocoa products. Consumption of cocoa and dark chocolate products has been associated with positive health outcomes including reduced onset of cardiovascular disease, inflammation, diabetes, obesity, and platelet disorders. Cocoa polyphenols, compounds increasingly associated with health benefits, are highly impacted by the variety and processing history of cocoa. However, the extent to which cocoa genetic variety and processing conditions impact polyphenol concentration and composition is not fully understood. Polyphenol characterization was conducted for five genetic varieties sourced from the USDA cocoa germplasm repository located in Mayagüez, Puerto Rico and it was found that genetic variance could contribute to differences in polyphenol concentration. The main focus in this thesis was to develop laboratory cocoa fermentation model system to facilitate cocoa research in laboratories distant from the cocoa growing region. This model system used dried, unfermented cocoa beans and simulated pulp medium as the starting material. The model system was effective in that it supported growth of the essential succession of coca fermenting microorganisms and generated similar chemical changes to those observed in on-farm cocoa fermentation. This model system could be further applied to test the impact of fermentation conditions on cocoa bean quality. The potential for using yeast starter cultures to improve control and consistency of cocoa fermentation was evaluated using the model fermentation system. Yeast inoculation improved fermentation rate, and polyphenol concentration in fermented beans was higher when cocoa fermentation was inoculated with the yeast strain Saccharomyces cerevisiae Lev B. Overall, the present work addressed the influence of cocoa variety selection and fermentation process conditions on the composition and concentration of polyphenols. These findings will contribute to continued efforts to develop cocoa products optimized for maximum health benefits.