Acute myocardial infarction (AMI) rat model was adopted to test the effect of AngioT in vivo. AngioT was directly injected into the ischaemic region of the left ventricle immediately after the ligation of left anterior descending artery. The densities of vessels in AngioT treated hearts were on average 3-4 folds higher compared with non-treated hearts after two and seven days post infarction. Using TUNEL method, approximately 3-fold lower numbers of apoptotic cardiomyocytes were detected in AngioT treated AMI hearts compared with controls. The infarcted volume estimated by Masson's Trichrome staining was significantly decreased in AngioT treated hearts (27.44%+/-7.34% vs. 39.53%+/-5.97%, p<0.05) compared with control hearts 14 days post infarction. Echocardiography demonstrated that left ventricular ejection fraction and fraction shortening in AngioT treated hearts were significantly improved by 10.4% and 22.3% compared to those in the control hearts 2-day post infarction (p<0.05). These improvements were maintained for 2-week post infarction. / Based on the analysis of rat angiogenesis specific superarray, VEGFb, VEGFc and FGF7, were found to be highly expressed in AngioT treated AMI hearts compared to the controls. The expression levels of survival related genes Bcl2 and Akt1 were increased to 3.3 and 2.8 folds respectively in AngioT treated AMI hearts compared with the controls (both p<0.05). Based on the signal transduction pathway finder superarray, Jak-Stat pathway activators, Interleukin 4 receptor and Interferon regulatory factor 1 (IL4R and IRF1), were found to be highly expressed in the AngioT treated AMI heart. / In conclusion, bioactive angiogenic factors (AngioT) were isolated from Geum. japonicum Thunb. Var. Chinense F. Bolle (GJ). Intra-cardiomuscular injection of AngioT had beneficial effects on the acute myocardial infarction. The underlying mechanisms might be related to the activation of Jak-Stat Pathway and over expressions of angiogenic factors and survival associated genes. The therapeutic properties of AngioT appear entirely novel and may provide a new dimension for therapeutic angiogenesis for the treatment of acute ischaemic heart disease. / In the present study, an angiogenic tannins fraction (AngioT) was isolated from Geum. japonicum Thunb. Var. Chinense F. Bolle using bio-assay guided strategy. AngioT increased the proliferation of human umbilical vein endothelial cell (HUVEC) in culture within 24-hour, 48-hour and 72-hour treatment in a dose-dependent pattern. The EC50 of AngioT on HUVEC was less than 25mug/ml. Conditional media from AngioT treated HUVEC stimulated the proliferation of HUVEC significantly greater than conditional media from non-treated HUVEC. Using 2-dimensional electrophoresis and MALDI-TOF, VEGFa was identified in the AngioT treated conditional medium. / Ischaemic heart disease remains the leading cause of morbidity and mortality in most countries. Severe ischaemia of myocardium induces myocardial infarction and results in an irreversible loss of myocardium. Restoration of coronary blood flow by rapid angiogenesis may offer a direct and effective therapeutic way to intractable ischaemic heart diseases. / Key words. ischaemic heart disease; myocardial infarction; therapeutic angiogenesis; Geum japonicum; apoptosis; tannins; VEGF; Jak-Stat pathway / Gu, Xuemei. / "Sep 2007." / Adviser: Peter Tong Chun Yip. / Source: Dissertation Abstracts International, Volume: 69-08, Section: B, page: 4615. / Thesis (Ph.D.)--Chinese University of Hong Kong, 2007. / Includes bibliographical references (p. 159-177). / Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Electronic reproduction. [Ann Arbor, MI] : ProQuest Information and Learning, [200-] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Abstract in English and Chinese. / School code: 1307.

L’étude des interactions entre système fourrager, santé et croissance des lapins contribuera à proposer de nouvelles pratiques pour des systèmes d’élevage cunicole alternatifs, tels que ceux en agriculture biologique (AB) incluant une gestion intégrée de la santé. Mais cette stratégie est limitée par le manque de références sur la cuniculture AB, i) sur l'alimentation au pâturage (ingestion de fourrages verts, qualités nutritionnelles pour les lapins, etc) ; et ii) sur les risques sanitaires notamment le parasitisme, identifié comme un frein important au développement de la cuniculture AB. L’emploi de plantes riches en tannins condensés (TCs) comme le sainfoin, permettrait de diminuer l’utilisation d’antiparasitaires comme cela a été montré chez les petits ruminants, tant pour la gestion des nématodes que celle des coccidies. Mais le potentiel d’activité des TCs chez le lapin, tout comme le niveau d’ingestion et les performances de croissance, restent à explorer, et pourra servir la cuniculture AB et conventionnelle. Ce travail de thèse a pour objectif i) d’étudier l’intérêt du sainfoin comme ressource pour l’alimentation du lapin, et pour ses propriétés antiparasitaires, ii) définir le niveau d’ingestion au pâturage des lapins et les conséquences sur la production et iii) d’évaluer, le risque parasitaire au pâturage pour des lapins en production. Nous avons montré qu’un aliment enrichi en sainfoin distribué à partir du sevrage, avec une teneur en tannins de 1,8% d’équivalent d’acide tannique, n’a pas réduit l’installation de L3s de Trichostrongylus colubriformis, ni la fertilité des vers adultes, mais a réduit le potentiel d’éclosion des oeufs (-27 points), contribuant à réduire l’infestation de l’environnement. Un aliment enrichi en sainfoin contenant 1,2% d’équivalent d’acide tannique, distribué aux mères et aux lapins en croissance a eu un effet coccidiostatique : l’excrétion oocystale fécale de lapins nourris avec un aliment enrichi en sainfoin a été réduite de 60% par rapport à ceux ayant reçus l’aliment témoin. Si la réduction de l’excrétion oocystale de l’espèce Eimeria magna n’a pas pu être démontrée, en revanche, la réduction d’oocystes toutes espèces confondues dans l’environnement pourrait contribuer à diminuer le risque de coccidiose en élevage. Comparé à la luzerne, le sainfoin, plus riche en lignines, a une forte concentration en énergie (11,2 MJ/kg) et en protéines digestibles (110 g/kg). Au pâturage, lorsque la quantité d’herbe offerte dépasse 85 g MS/kg0,75, il semble que l’ingestion d’herbe soit régulée d’une part lorsque la teneur en énergie digestible de l’herbe dépasse 9 MJ/kg (régulation chémostatique), ou d’autre part si la teneur en lignocellulose (ADF) dépasse 350 g d’ADF/kg (régulation physique : encombrement digestif). Mais la quantité d'herbe disponible dépasse rarement 85 g MS/kg0,75. C’est-à-dire que dans la majorité des cas, une surface pâturable de 0,4 m² (minimum réglementaire) n’a pas permis de combler la capacité d’ingestion et les besoins énergétiques de lapin en croissance. De plus, si l’offre d’herbe est limitante, le lapin ne peut pas exprimer une préférence alimentaire vers des plantes plus jeunes et riches en protéines. La limitation de l’ingestion de protéines a aussi pour conséquence de réduire les potentialités de croissance des lapins et d’allonger la période d’engraissement. Au cours de trois saisons successives de pâturage (université de Perpignan), la pression parasitaire (nématodes et coccidies) a augmenté, avec des infestations par Trichostrongylus sp. et de Graphidium strigosum. Si le délai d'attente entre deux pâturages n’a pas eu d’effet visible sur l’infestation par des nématodes, cela influerait le niveau d’infection par les coccidies. Nos travaux établissent l’intérêt de l’incorporation du sainfoin dans l’alimentation du lapin et ouvrent des perspectives pour établir des pratiques innovantes et bénéfiques à la production cunicole biologique et conventionnelle. / The study of the interactions between grazing systems, health and growth in organic rabbit farming (ORF) will contribute to propose new practices including integrated management of health. However, this strategy is limited by the lack of references on ORF and in particular on rabbit intake of green fodder when grazing, nutritional qualities of forages for rabbits, the supply of grass in relation with animal’s density (stocking rate), the related parasitic challenge, etc. The acquisition of such references is a key issue for the development of ORF. The use of plants rich in condensed tannins (CTs), such as sainfoin, could reduce the use of antiparasitic agents, as shown in small ruminants, both for the management of nematodes and for coccidia. However, the potential for CTs containing resources in rabbits, as well as the intake levels and the effects on growth performances, remain to be explored in both OF and conventional rabbit breeding. The aim of this thesis was (i) to investigate the interest of sainfoin as a resource for rabbit feeding and its antiparasitic properties, (ii) to define the intake level in grazing rabbit and the consequences on production and (iii) to evaluate the parasitic risk associated with pasture for rabbits’ production. We have shown that a sainfoin-enriched diet distributed from weaning, with a feed containing 1.8% of tannic acid equivalent, did not reduce neither the establishment of Trichostrongylus colubriformis L3s, nor the fertility of adult worms. In contrast, the development of nematode’s egg to infective larvae was compromised and may reduce the risk of environmental contamination. Besides, a sainfoin-enriched diet containing 1.2% tannic acid equivalent, distributed to does and growing rabbits, had a coccidiostatic effect. With an overall fecal oocyst excretion in rabbits fed with sainfoin reduced by 60% compared to control diet. Although, the pathogenic species Eimeria magna was not concerned by these decrease, such a general reduction in oocyst excretion in the environment could help to reduce on long term the risk of coccidiosis in the rearing units. Sainfoin could constitute a real alternative to dehydrated alfalfa, since it has high digestible energy (DE, 11.12 MJ DE/kg), and protein (110 g/kg), and provide a high supply of lignins. At grazing, when the herbage allowance exceeded 85 g DM/kg0,75, it appeared that the herbage intake was limited by an increase in the DE content when it exceeds 9 MJ / kg, or else by a lignocellulose content higher than 350 g of ADF/kg. However, herbage allowance, rarely exceeds 85 g DM/kg0,75. In other words, in the majority of cases access to a 0.4 m² grazing area did not allow sufficient supply to reach the intake capacity and energy requirements of the rabbits. In addition, rabbits with limited supply cannot express their dietary preference to young and high protein plants. Limiting the intake of proteins also reduced the growth potential of rabbits and extended the fattening period. During the first three grazing seasons (University of Perpignan), the parasitic pressure (nematodes and coccidia) increased, particularly with cases of Trichostrongylus sp. and Graphidium strigosum. While rotation rhythms had no visible effects on nematode infestation, it had an influence on coccidian infection. This work enabled us to establish the advantage of sainfoin enriched diet for the rabbit. It gives prospects to further research projects regarding the establishment of beneficial and innovative practices for organic and conventional rabbit breeding.

Les tannins sont des polymères de polyphénols présents en quantité significative dans le vin rouge, et responsables de l’astringence et de l’amertume. L’astringence est une sensation de sècheresse et de rugosité en bouche résultant d’une forte interaction entre les tannins et les protéines de la salive impliquées dans la lubrification de la cavité buccale. L’amertume, quant à elle, est un goût stricto sensu résultant de l’interaction spécifique des tannins avec les récepteurs du goût situés dans les papilles linguales. Des études récentes ont montré que les tannins sont susceptibles d’interagir avec les lipides. Or les lipides sont présents lors de la dégustation d’un vin comme composants des membranes buccales ou des aliments gras. Cependant, le rôle des lipides dans les perceptions sensorielles d’un vin n’est pas bien connu d’un point de vue œnologique. L’objectif de cette thèse était d’étudier au niveau moléculaire les interactions tannin-lipide, pour mieux comprendre leur rôle dans les propriétés gustatives du vin. Le présent travail décrit l’effet de deux entités représentatives des tannins du vin : un monomère, la catéchine, et un dimère, la procyanidine B1, sur deux modèles lipidiques. Le premier modèle est un modèle membranaire représenté par des vésicules multilamellaires composées de POPC/Cholestérol (70/30), qui mime la composition lipidique des membranes buccales. Le deuxième modèle est une émulsion huile dans l’eau (H/E) stabilisée par le DMPC, qui mime les gouttelettes lipidiques présentes dans les aliments gras. L’organisation et la dynamique des lipides composant ces deux modèles ont été étudiées par la spectroscopie RMN (1H, 2H, 13C) en présence et en absence des deux entités de tannins. Leur localisation dans les membranes lipidiques a également été explorée, de même que leur affinité pour les lipides avec la détermination des constantes d’association tannin-lipide. Les résultats ont mis en évidence un effet fluidifiant des tannins à la fois sur le modèle de membranes buccales et sur le modèle de gouttelettes lipidiques. On a démontré que cet effet de désordre est lié à la nature chimique des tannins, ainsi qu’à leur position dans la membrane. De plus, les résultats sur l’affinité tannin-lipide sont en faveur d’une compétition avec les protéines salivaires. En outre, les résultats de biophysique se sont avérés conformes avec ceux d’une analyse sensorielle menée en parallèle qui a révélé que les aliments gras sont susceptibles de diminuer l’astringence du vin. Ces travaux montrent l’impact des composés phénoliques sur l’ordre membranaire et soulignent pour la première fois un rôle potentiel des lipides sur le goût du vin. D’une part, les interactions tannin-lipide, en perturbant l’environnement lipidique des récepteurs du goût enchâssés dans les membranes buccales, pourraient affecter la fonctionnalité du récepteur et son interaction avec les tannins, et donc l’amertume. D’autre part, une éventuelle compétition entre les interactions tannin-lipide et tannin-protéine de la salive pourrait diminuer l’astringence durant la dégustation d’un vin. Dans le domaine de l’œnologie, cette thèse vient étayer le ressenti des dégustateurs à savoir la modification du goût du vin due aux aliments et ouvre de nouvelles perspectives dans le cadre de l’association mets-vins. / Tannins are polyphenol polymers present in significant amounts in red wine responsible for astringency and bitterness. The former is a tactile perception involving dryness and roughness in the mouth due to the interaction between tannins and saliva proteins and the latter is a primary taste due to the interaction between tannins and taste receptors in taste buds. Tannins are now known to also interact with lipids. Although not present in wine, lipids are yet present during tasting in the oral membranes of tasters and in fatty foods when wine is consumed during a meal. However, although the influence of lipids is well known to wine tasters through food pairing, there is no scientific evidence to support this hedonic feeling. The aim of the thesis is to study tannin-lipid interactions at molecular level in order to better understand their implication in wine gustative properties. The present work describes the effect of the main representative grape tannin subunits, the catechin monomer and the B1 dimer, both on a model of oral membranes and food fat globules. They are represented by a dispersion of POPC/cholesterol multilamellar vesicles and a olive oil in water emulsion stabilized by DMPC as emulsifier, respectively. The organization and dynamics of the lipids composing these two models were investigated by solid-state NMR spectroscopy (1H, 2H, and 13C) in the absence and the presence of the two tannin subunits. The affinity of tannins for lipids was also explored by the determination of the thermodynamic association constant. The results pointed out a fluidizing effect of tannins both on the membrane model, as previously shown on a simpler membrane model, and on the emulsion lipid droplets. The disorder caused by tannins was shown to be related to their location in the lipid structure depending on the tannin chemical nature. Moreover, the strength of the interaction between tannins and membrane lipids was revealed to be in the same order of magnitude of that between tannins and saliva proteins. In addition, the biophysical results were in accordance with those of a sensory analysis led in parallel that revealed that fatty foods are prone to decrease wine astringency. These pioneering works shows the impact of phenolic compound on membrane order and highlight for the first time the potential role of the tannin-lipid interactions on wine taste. On the one hand, by disrupting the lipid environment of taste receptors embedded in oral membranes, tannin-lipid interactions could affect the receptor functionnality and therefore the interaction with tannin molecules, so bitterness. On the other hand, the existence of a possible competition between lipids and saliva proteins for interacting with tannins during tasting could reduce astringency.

Les paramètres thermodynamiques et les cinétiques d’interactions entre des polyphénols (des procyanidines et des anthocyanes) et des fractions pectiques ont été déterminés par des méthodes physico-chimiques en solution et sur support solide. Les expériences ont été réalisées avec les anthocyanes majoritaires de cassis ou des procyanidines de type B (extraites de pomme) avec différents degrés de polymérisation et des fractions pectiques issues de pectines de pomme présentant divers degrés de méthylation et différentes chaines latérales d’oses neutres.Les interactions entre les anthocyanes et les pectines sont influencées par les substituants osidiques et le nombre de groupements hydoxyles du noyau B des anthocyanes ainsi que par la composition des fractions pectiques. Les constantes d’affinité entre les pectines et les procyanidines en solution sont les plus élevées quand le degré de polymérisation des procyanidines et le degré de méthylation des pectines sont les plus élevés. De plus, le niveau de ramification des pectines limite leur association avec les procyanidines. Ces interactions impliquent à la fois des liaisons hydrogènes et des interactions hydrophobes. Après les modifications chimiques et l’immobilisation des procyanidines sur une surface, les unités de résonance obtenues par résonance plasmonique de surface entre la (-)-épicatéchine ou le dimère DP2 et les pectines de pomme sont similaires à celles obtenues avec la protéine riche en proline IB5, mais plus faible qu’avec la sérum albumine bovine. / Thermodynamical parameters and kinetics of interactions between polyphenols (procyanidins and anthocyanins) and pectic fractions were defined by physico-chemical methods in solution and with a solid support. Experiments confronted major anthocyanins presents in blackcurrant or B-type procyanidins from apple with various degrees of polymerization to pectic fractions from apple pectins presenting different degree of methylation and different neutral sugar side chains.Interactions between anthocyanins and pectins are influenced by the glycosyl substituent and the number of hydroxyl groups in the B-ring of anthocyanins, and by the composition of pectins. The affinity constant of procyanidins – pectins interaction in solution are the highest when both the procyanidins degree of polymerization and the pectins degree of methylation are the highest. Moreover, the ramification state of pectins limits their association with procyanidins. Those interactions are due to hydrogen bonds and hydrophobic interactions.After chemical modifications of procyanidins and immobilization on a solid support, the resonance units obtained by surface plasmon resonance between (-)-epicatechin or dimer B2 and apple pectins are similar to those obtained with the prolin-rich protein IB5 and lower than with bovine serum albumin.

Les tannins sont des extraits chimiques présentant une alternative naturelle aux produits chimiques de type phénol ou résorcinol issus du pétrole. Ils sont déjà utilisés industriellement pour la synthèse d’adhésifs utilisés pour la fabrication de produits à base de bois tels que les panneaux de particules et les contreplaqués, mais généralement en utilisant toujours du formaldéhyde comme durcisseur pour améliorer leur pouvoir collant. Les premiers types de produits développés dans cette thèse sont des résines adhésives pour fabriquer des panneaux de particules, synthétisées à partir de tannins de Pin Radiata et d’autres aldéhydes moins nocifs obtenus en faisant réagir de l’huile de tournesol avec de l’ozone puis avec agent réducteur. L’étude s’est portée sur les conditions de l’ozonolyse, dont les paramètres étaient nombreux, et sur les analyses des huiles obtenues, puis sur la synthèse de résines à partir de solutions de tannins. Certaines de ces résines ont été utilisées pour fabriquer des panneaux de particules qui ont été testés en cohésion interne sèche. Puis le sujet s’est orienté vers le développement de mousses rigides à base de tannins et d’alcool furfurylique dans le cadre d’un projet industriel multi-matériaux. Plusieurs formulations ont été testées pour atteindre les objectifs d’un cahier des charges, dont la finalité était d’obtenir en premier lieu des panneaux de mousse homogènes de basse densité avec de bonnes propriétés mécaniques. L’innovation apportée a été de produire ces mousses dans une presse à panneaux. Enfin, une nouvelle façon de fabriquer des polymères polyuréthanes à base de tannins a été testée en utilisant des produits chimiques moins nocifs en remplacement des isocyanates habituellement utilisés dans l’industrie. La synthèse s’est fait d’abord par une étape de carbonatation des groupes hydroxyles des tannins, puis par une réaction avec l’hexamethylènediamine. Les analyses spectrométriques montres que les produits obtenus possèdent bel et bien des liaisons uréthanes. L’étude débouché sur l’exploration de l’utilisation de tannins réagis avec de l’ammoniac pour le remplacement de l’hexaméthylènediamine afin d’obtenir de nouveaux polyuréthanes « non isocynates » avec encore plus de produits biosourcés et plus respectueux de l’environnement / Tannins are chemical extracts which represent a natural alternative to chemicals as phenol or resorcinol which are from oil resources. They are already used industrially for the synthesis of adhesives used for the manufacture of wood-based products such as particleboards and plywood, but generally always used with formaldehyde as a hardener to improve their mechanical strengths. The first products developed in this thesis are adhesive resins for the manufacture of particleboards, synthesized from Radiata Pine tannins and less hazardous aldehydes obtained by reacting sunflower oil with ozone then with a reducing agent. The study has focus first on the conditions of the ozonolysis, whose parameters are numerous, and the analysis of the oils obtained, then on the synthesis of resins with solutions of tannins. Some of these resins have been used to manufacture particleboards that have been tested in dry internal bond strength. Then the subject has been oriented toward the development of rigid tannin/furanic-based foams as part of an industrial multi-material project. Several formulations have been tested to achieve the objectives of a specification, whose first purpose is to obtain foam panels homogeneous, low in density and with good mechanical properties. The innovation has been to make these foams in a particleboard press. Finally, a new route to manufacture tannin-based polyurethane polymers has been tested by using less hazardous chemicals in replacement of isocyanates, conventionally used in the industry. The synthesis is made by a first carbonatation step of tannins hydroxyl groups and then by a reaction with hexamethylenediamine. Spectrometric analyzes have shown that the products obtained do have urethane linkages. The study led to the exploration of the use of tannins reacted with ammonia to replace hexamethylenediamine to obtain new "non isocynates" polyurethanes with more bio-based and environment-friendly chemicals

Les marcs de raisin issus de la viniculture constituent un gisement de ressource naturelle abondant, sous valorisé et riche en polyphénols. Une méthode d?extraction de ces tannins condensés a été développée et optimisée en vue d'une application en adhésif pour les produits dérivés du bois. Les tannins ont été caractérisés par Résonance Magnétique Nucléaire, spectroscopie Infra Rouge (FTIR), spectrométrie de masse (MALDI TOF) et Analyse Thermomécanique (TMA). Des panneaux de particules ont été fabriqués à l'aide des colles élaborées en laboratoire et testés suivant les normes françaises en vigueur, plusieurs dépassent largement les valeurs seuils / The extraction of condensed tannins from grape pomace was examined using water medium in the presence of different bases as catalyst (NaOH, Na2CO3, NaHCO3 and Na2SO3). Two different extraction processes and the influence of 4 parameters (i.e., temperature, reaction time, chemical reagents and concentration of the chemical reagents) on the tannin extracts yields and properties were studied. The tannin fractions were characterized by Fourier transform-infrared spectroscopy, thermogravimetric analysis, carbon nuclear magnetic resonance spectroscopy and matrix assisted laser desorption ionization time of flight mass spectrometry ( MALDI-TOF ). It was demonstrated that it is possible to extract reactive tannins from grape pomace in basic medium. The tannin extracts obtained by precipitation in acidic conditions display a high phenolic content (Stiasny number higher 95%) but low water solubility and low reactivity toward formaldehyde because of the formation of catechinic acid phlobatannins during the acidification step. The tannins extracts obtained by lyophilization of the liquid, despite of their lower phenolic contents, displayed promising properties for adhesive applications. The optimum temperature of the extraction process was 100 °C, reaction time was 120 min; the best concentration of reagent was 10% (w/w). It was also shown that addition of sulphite ions during the extraction step improved the process :y the introduction of a sulfonic group through sulfitation increased both tannin solubility and reactivity as a result from the opening of the heterocyclic ring during extraction The structure of grape pomace sulfited tannin extracts did not present noticeable discrepancy exception of the ratio of the opening pyran ring which was different as a function of the catalyst used. The opening of pyran ring during the tannins extraction seem to be more important by using Na2CO3 and NaHCO3 than when using NaOH. The FT-IR bands assigned to aromatic ring vibration and carbonyl groups were stronger and no bands attributed to sulfited groups were tested detected in acidified tannins compared to lyophilized tannins. The TG2 results showed that the weight lost of these tannin extracts mainly composed of two steps. The first step starts from room temperature to 200 °C. It is attributed to the mass lost of water and some easy-degraded small low molecules. The secondly steps which is the mainly degradation step of tannin extract samples from 200 to 400 °C. The results from 13C-NMR showed that condensed tannin extracts from grape pomace were consistent with dominant procyanidin units with a minor amount of prodelphinidin units that are linked together by a C4-C8 bond. Relatively low carbonhydrate and high catechinc acid content was observed in acidified tannins compared to lyophilized tannins. It was shown by MADI-TOF experiments that grape pomace tannin extracts are mainly composed of flavoinoid oligomers up to 6 repeating units in lyophilized tannins and 10 repeating units in acidified tannins respectively, with dominant procyanidin units. A small proportion of substitution with glucose and gallic acid was detected in procyanidin units of polyflavonoid oligomers. The degree of polymerization of acidified tannins is higher than lyophilized tannins. Two different formulations (nonfortified tannin adhesive and fortified with addition of 20% of polymeric 4, 4'-diphenyl methane diisocyanate (p-MDI)) were used to press one layer wood particle board. It was shown that the nature of the extraction reagent (NaOH, Na2CO3 or NaHCO3) greatly impacts the properties of the resins and the mechanical properties of the panel internal bonding strengths. The particleboards bonded by the tannins extracted using Na2CO3 as catalyst give the best performance and were good enough to pass relevant international standard specifications for interior grade panels. Formaldehyde emission of these panels was below the European Norm requirements (<= 6.5 mg/ 100g panel)

Doctor of Philosophy / Department of Food, Nutrition, Dietetics, and Health / Brian L. Lindshield / Iron, vitamin A, and protein inadequacies are common in food-aid receiving countries, and maximizing nutrient intake and bioavailability are essential treatments. Fortified-blended foods (FBFs), are food-aid micronutrient-fortified legume-grain porridges distributed worldwide. FBFs have not consistently, effectively treated undernutrition, and it has been suggested that formulation and processing changes could improve their nutritional quality. Sorghum is a well suited FBF commodity, but high concentrations of ‘antinutritional’ tannin and phytic acid have limited its adoption. Iron bioavailability adaptation may be possible after long-term antinutritional factor consumption, but adaptive mechanisms are not well understood. In rats, salivary proline-rich proteins (PRPs) have been found to chelate tannins to improve iron bioavailability, this could be true for people as well. Several research design methods were employed to summarize FBF quality outcomes and the effect of tannin and phytic acid consumption on iron bioavailability. Extruded sorghum and corn FBFs were developed; protein quality, iron, and vitamin A outcomes were compared with a non-extruded corn-soy blend (CSB+) in rats. A narrative literature review and meta-analysis were conducted to determine tannin’s antinutritional effects on iron bioavailability, and the potential for adaptation through salivary PRPs. Two clinical trials examined the effect of long-term tannin or phytic acid consumption on iron bioavailability, salivary protein production, and correlations between PRPs and iron bioavailability. There were no differences between iron (hepatic iron 207-300 µmol/g *100), vitamin A (hepatic retinol 423-585.5 ng/mg), or protein quality (caloric efficiency: 101.3-113.3 g/kcal*100) between extruded FBFs regardless of commodity in rats. Compared to extruded FBFs, CSB+ caloric efficiency (49.0 ± 2.2 g/kcal*100) and growth (96.3 ± 3.4g vs. 208.6-236.6) were significantly reduced. A literature review suggested that there were differences in acute meal and long-term iron bioavailability with tannin consumption; tannic acid inhibited iron availability, while food-tannins did not. Meta-analysis suggested that tannin-PRP binding could protect iron bioavailability, that long-term tannin consumption did not significantly affect hepatic iron or non-heme iron absorption respectively in rats (d = -0.64-1.84; -2.7-0.13), and that PRP expression in rats during tannin consumption was correlated with improved iron bioavailability. There were no reductions in iron bioavailability or status based on long-term tannin (ps > 0.126) or phytic acid (ps > 0.08) consumption clinically, but basic PRP and cystatin subtypes were significantly correlated with improved iron bioavailability during tannin (ps < 0.03) and phytic acid (ps < 0.02) consumption. In vitro, it phytic acid-PRP binding did not occur, but phytic acid did specifically bind with cystatin SN, a non-enzymatic salivary protein. In conclusion, FBF formulation changes may improve protein quality, and provide needed macronutrients to food-aid receiving areas. Despite this, this research did not suggest that antinutritional factors affected iron bioavailability. In support of this finding, literature, and clinical studies presented here suggest that salivary proteins, including PRPs and cystatin, may serve as adaptive protective mechanisms against phytic acid and tannin consumption, and that further research may be warranted before further recommendations for their removal from food-aid are made.

Master of Science / Department of Animal Sciences and Industry / KC Olson / Our objective was to determine the effects of supplemental corn steep liquor (CSL) on voluntary selection of sericea lespedeza (SL) by beef cows grazing native tallgrass range. Dietary botanical composition of cows (n = 145; initial BW = 579 ± 91 kg) was evaluated during a 150-d grazing period (5/1 to 10/1). Native pastures (n = 9; 50 ± 17 ha) infested by SL (average SL biomass = 37% of total forage biomass) were assigned randomly to 1 of 2 treatments: grazing by unsupplemented cows or grazing by cows supplemented with CSL (1.79 kg DM • cow-1 • d-1; 45% DM, 34% CP). Cows were assigned randomly to treatment and pasture (stocking rate = 0.5 ha/AUM). Concentration and protein-binding capacity of condensed tannins (CT) in SL were measured monthly. Fecal samples were collected from each cow on 6/1, 7/1, 8/1, 9/1, and 10/1. Herbivory of SL was estimated along line transects in October. Plant fragments in fecal samples were quantified via a microhistological technique; fragment prevalence in fecal material was assumed to equal botanical composition of the diet. Concentration and protein-binding capacity of CT in SL were greatest (P < 0.01) on 8/1 and 9/1, respectively. The proportion of individual SL plants showing evidence of herbivory tended to be greater (P = 0.09) on pastures grazed by supplemented cows compared to pastures grazed by unsupplemented cows (94 vs. 80% of SL stems, respectively). Prevalence of SL in beef cow diets was influenced (P < 0.01) by CSL supplementation and by month. Prevalence of SL in beef cow diets was not different (P ≥ 0.35) between treatments when concentration and protein-binding capacity of CT were relatively low (6/1, 7/1, and 10/1). In contrast, supplemented cows selected more (P < 0.01) SL than unsupplemented cows when concentration and protein-binding capacity of CT were greatest (8/1 and 9/1). We interpreted these data to suggest that voluntary selection of SL by beef cows was inversely related to concentration of CT; moreover, supplemental CSL stimulated voluntary selection of SL during periods of high CT concentration. Supplemental CSL did not influence selection of other plant species that were monitored.

Diversas plantas com atuação no trato gastrointestinal - ricas em taninos e antraquinonas - ainda deixam muitas dúvidas quanto a sua caracterização estrutural, o seu mecanismo de ação e toxicidade. O presente trabalho objetivou o estudo químico e biológico de duas plantas utilizadas para doenças relacionadas ao sistema gastrointestinal: barbatimão (Stryphnodendron rotundifolium), rico em procianidinas e taninos (utilizada para combater gastrite); e cáscara sagrada (Rhamnus purshiana), rica em antraquinonas (utilizada para constipação). Inicialmente foi realizado um estudo químico com as plantas envolvendo isolamento e caracterização de componentes do extrato por RMN (1H, 13C, DEPT e NOESY), espectrometria de massas (MS), infravermelho e dicroísmo circular vibracional (VCD). Essas substâncias foram utilizadas para a padronização dos extratos utilizados em estudo biológico. O estudo biológico envolveu testes in vitro para avaliar a toxicidade de diferentes componentes do extrato de cáscara sagrada e teste in vivo com barbatimão, visando, através do tratamento de animais e remoção do estômago para estudo em microscopia confocal, entender o mecanismo de proteção gástrica. Com o desenvolvimento do trabalho foi possível desenvolver novas metodologias para a identificação de antraquinonas utilizando espectrometria de massas e provar a veracidade de regras empíricas utilizadas para a determinação da configuração absoluta de moléculas dessa classe. Os testes in vitro com antraquinonas isoladas e padrões comerciais indicam a possibilidade de aprimoramento dos perfis químicos utilizados para a produção de medicamentos a base de extratos de cáscara sagrada, visando à redução de toxicidade. Quanto ao barbatimão, o estudo químico possibilitou a aplicação da técnica de VCD para a diferenciação de procianidinas diasteroisomericas. Os estudos in vivo confirmaram a eficácia do uso de taninos para combater gastrite através da formação de um revestimento protetor. / Several plants with action in the gastrointestinal tract - rich in tannins and anthraquinones - cause many doubts concerning their chemical characterization, mechanism of action and toxicity. The present work aimed to develop a chemical and biological study of two plants used for gastrointestinal system related diseases: barbatimão (Stryphnodendron rotundifolium), rich in procyanidins and tannins (used to treat gastritis); and cascara sagrada (Rhamnus purshiana), rich in anthraquinones (used for constipation). Initially, a chemical study with plants was performed comprising isolation and characterization of components of the extract through NMR (1H, 13C, DEPT and NOESY), mass spectrometry (MS), and infrared and vibrational circular dichroism (VCD). These substances were used for extract standardization used in biological studies. The in vitro tests were performed in order to evaluate the toxicity of different components of the cascara extract. The in vivo tests with barbatimão aimed to better understand the mechanism of gastric protection, by treating animals with the extract, removing their stomachs, and analyzing them with fluorescence microscopy. Through this work we were able to develop new methods for identifying anthraquinones by mass spectrometry and to prove empirical NMR rules used to determine the absolute configuration of molecules of this class. The tests with isolated anthraquinones indicated the possibility of improving chemical profile to further reduce cascara sagrada toxicity. The barbatimão chemical study allowed the application of VCD technique for the differentiation of diasteroisomeric procyanidins, and confirmed the effectiveness of using tannins for gastritis treatment through the formation of a protective coating.

Objetivou-se com o presente estudo avaliar o efeito dos aditivos naturais, óleo essencial de orégano e extrato taninífero de acácia negra, na alimentação de cordeiros confinados em terminação sobre o desempenho produtivo e qualidade da carne. Foram utilizados 36 ovinos machos, não castrados, provenientes de cruzamento Dorper × Santa Inês × Texel, com 90 dias de idade, e peso vivo inicial médio de 27,22 &amp;plusmn; 2,78 kg. O experimento teve duração total de 53 dias. O delineamento experimental utilizado foi em blocos inteiramente casualizados, utilizando como critério de bloqueamento o peso ao desmame, onde os animais foram distribuídos em três tratamentos, com doze repetições. Todos os tratamentos foram compostos por uma dieta padrão, sendo distintos apenas em relação aos aditivos utilizados: 1) dieta controle; 2) dieta controle com óleo essencial de orégano (1 ml/kg de concentrado) e 3) dieta controle com extrato de taninífero de acácia negra (5% MS da dieta). A dieta foi formulada com proporção volumoso: concentrado de 30:70. Os cordeiros foram destinados ao abate com aproximadamente 143 dias de idade, e peso vivo médio final médio de 42,81 &amp;plusmn; 3,97 kg. O consumo e digestibilidade dos nutrientes e o desempenho produtivo (ganho de peso total, ganho médio diário e peso e rendimento de carcaça) dos cordeiros alimentados com óleo essencial de orégano foi semelhante ao dos alimentados com a dieta controle. No entanto, os cordeiros alimentados com extrato taninífero de acácia apresentaram menor consumo de MS (CMS), e redução de 19%, 20% e 50% da digestibilidade aparente total da MS, MO e FDN, respectivamente, em relação aos cordeiros alimentados com a dieta controle. Consequentemente, os cordeiros alimentados com extrato taninífero de acácia apresentaram menor ganho de peso e rendimento de carcaça quente e fria do que os alimentos com a dieta controle. O óleo essencial de orégano deixou a carne mais luminosa, no entanto, apresentou uma tendência de deixar a carne mais dura. O óleo essencial alterou a composição centesimal da carne e a concentração de TBARS, e aumentou o teor do ácido graxo DHA em 13% em relação ao tratamento controle. O extrato de acácia negra também aumentou a luminosidade da carne, porém, reduziu em 4% o teor de PB da carne e a concentração de TBARS em 28% em relação ao tratamento controle. O extrato taninífero tendeu a aumentar o teor de ácido vacênico e de ômega 3. Adicionalmente, o extrato taninífero aumentou em 27% o total de CLA, quando comparado ao controle e óleo essencial de orégano. Pode-se concluir com o presente estudo que a inclusão de óleo essencial de orégano na dieta de cordeiros em terminação não altera o desempenho produtivo durante o confinamento e aumenta a luminosidade da carne, apensar de deixar a mesma mais dura; enquanto que a utilização de extrato taninífero de acácia reduz o desempenho produtivo, porém melhora a qualidade da fração lipídica e antioxidante da carne. / TThe present study aimed to evaluate the effect of natural additives, oregano essential oil and acacia taniniferous extract, in the diet of feedlot lambs on productive performance and meat quality. Thirty-six male lambs (not-castrated) from crossbreeding Dorper × Santa Inês × Texel, weaned, averring 90 d of age and 27 &amp;plusmn; 2.8 kg body weight were used through 53 d of feedlot period in a randomized block design, were weigh at weaning was the block criteria. The lambs were distributed into three treatments with 12 repetitions. All treatments were based on a regular diet (control), where natural plants extracts were designated to following treatments: 1) control (without additive), 2) control diet + oregano essential oil (1 ml/kg of concentrate), 3) control diet + acacia taniniferous extract (5% DM). Diets were formulated as 30:70 forage:concentrate ratio. Lambs were designated to slaughter averring 143 days of age and 42,81 &amp;plusmn; 3,97 kg body weight. The nutrient intake and digestibility and productive performance (total body weight gain, average daily body weight gain and production and carcass yield) of lambs fed oregano essential oil were similar to those fed control diet. However, lambs fed acacia taniniferous extract had lower DM intake (DMI), and reduced by 19%, 20% and 50% the total apparent digestibility of DM, OM and NDF, respectively, compared to lambs fed control diet. Meat of lambs fed oregano essential oil was more luminous, however tended to be stronger. Oregano essential oil changed the centesimal composition of meat and TBARS, but increased concentration of DHA fatty acid by 13% compared to control treatment. Acacia taniniferous extract also increased the luminosity of meat, but reduced by 4% meat CP content and TBARS concentration by 28%, in relation to control treatment. Acacia taniniferous extract tended to increase vaccenic acid and omega 3 content. Additionally, taniniferous extract increased by 27% the total CLA, compared to control and oregano essential oil. We can conclude that the inclusion of oregano essential oil in the diet of feedlot lambs does not change productive efficiency and increase the luminosity of meat, although results in stronger meat; whereas the inclusion of acacia taniniferous extract reduce productive performance, but increase the quality of lipid fraction and antioxidant of meat.