Cette étude a pour objectif d'apporter des éléments de réponse à la compréhension de la nature et de l'origine des effets biologiques des nanoparticules (NP) de boehmite en réalisant des tests de contacts in vitro sur des macrophages de culture (RAW 264.7), les macrophages étant la cible cellulaire privilégiée au niveau du tractus respiratoire. Différents domaines de la réponse cellulaire en relation avec les propriétés physico-chimiques des particules ont été étudiés. Dans une première partie l’activité biologique a été mesurée sur des NP de boehmite puis après broyage ou dispersion permettant d’observer en particulier l’influence de la taille des particules dans la réponse cellulaire.Les résultats soulignent que les particules industrielles de boehmite sont principalement caractérisées par une activité modérée au niveau de la réponse inflammatoire, mais sans effet cytotoxique ou sans stress oxydant significatif. Des différences en fonction de la taille des particules ont été observées sur les paramètres inflammatoire et cytotoxique.Toutefois, les mesures des biomolécules libérées dans les surnageants de culture cellulaire peuvent être biaisées par l’adsorption de ces biomolécules sur les NP en présence. Ainsi, ce mécanisme d'adsorption doit être pleinement compris pour éviter une interprétation erronée des données obtenues. Dans un second temps l’objectif a donc été d’élaborer une méthodologie afin d’évaluer les interactions organo-minérales en terme d’affinité et de quantité de biomolécules adsorbées sur les nanoparticules à l’équilibre thermodynamique et de déterminer une loi de correction. Dans ce contexte nous nous sommes intéressés en particulier au TNF-α. / This study aimed at a better understanding of the biological effects of boehmite nanoparticles by carrying out in vitro tests using a macrophage-like cell line (RAW 264.7), as macrophages represent the first target cells in the respiratory tract. Different cellular responses: inflammation (evaluated by the release of TNF-α), cell death (assessed by the release of LDH), and oxidative stress (production of ROS) were studied in relation to physico-chemical properties of the particles. In the first part of this work, the biological activity was measured using boehmite nanoparticles obtained either from industrial environment or by hydrothermal synthesis, and after grinding or dispersion that enabled us to observe the influence of particle size on the cellular response.The results point out that industrial boehmite particles are mainly characterized by a moderate pro-inflammatory activity, no cytotoxic effect and no significant oxidative stress. Differences depending on the size of the particles were observed on the inflammatory and cytotoxic parameters.However, the assessment of biomolecules released into cell culture supernatants may be biased by the adsorption of biomolecules on the nanoparticles. Thus, the adsorption mechanism must be fully understood to avoid misinterpretation of the results. Therefore, the second aims of this work were to develop a methodology to assess the organo-mineral interactions in terms of affinity and quantity of biomolecules adsorbed on the nanoparticles at the thermodynamic equilibrium and to determine a correction law. We focused more specifically our study on TNF-α.

Afin de dépolluer et de réhabiliter les sols contaminés par des hydrocarbures aromatiques polycycliques (HAP), différentes techniques ont été développées durant les dernières décennies, mais leur impact sur les fonctions du sol est rarement pris en compte. L'objectif de ce travail était d'évaluer l'impact des traitements par oxydation chimique sur différents paramètres physico-chimiques et biologiques de terres contaminées aux HAP et de proposer des solutions pour restaurer le fonctionnement biologique des terres oxydées. Les différentes expérimentations de laboratoire et de terrain ont montré que l'efficacité des réactions d'oxydation chimique (réaction de Fenton et persulfate activé) sur la teneur en HAP dépendait de la disponibilité de la pollution et des propriétés physico-chimiques des terres. Par ailleurs, des processus d'acidification ont été observés sur des terres non carbonatées, altérant fortement et durablement les paramètres physico-chimiques et biologiques. Cette altération était plus forte après utilisation du persulfate activé, engendrant ainsi des pertes en éléments traces métalliques dans les eaux de percolations. L'apport de matériaux susceptibles d'augmenter le pH a permis la restauration des activités biologiques et la capacité des terres à supporter la végétation lorsque les terres ont fortement été acidifiées. L'association de traitements d'oxydation chimique et de refonctionnalisation pourrait présenter un moyen de réhabilitation efficace. Un indice, dont l'objectif était de représenter l'état du sol en fonction du poids accordé à différents paramètres, a été proposé et a permis d'évaluer les variations induites par les traitements d'oxydation et les procédés de refonctionnalisation / In order to clean up polycyclic aromatic hydrocarbon (PAH) contaminated soils, various techniques have been developed during the last decades, but their impact on soil functions was rarely considered. The objective of this study was to assess the impact of chemical oxidation treatments on different physico-chemical and biological parameters of PAH contaminated soil and to propose solutions to improve the biological functioning of oxidized soil. Various laboratory and field experiments have shown that the efficiency of chemical oxidation reactions (Fenton reaction and activated persulfate) on PAH concentrations depended on pollutant availability and soil physico-chemical properties. Besides, intense acidification processes have been observed on uncarbonated soil with a strong impact on soil physico-chemical and biological parameters. Moreover, this alteration was higher with activated persulfate treatment, leading to the loss of trace metals in water percolation. In the strongly acidified soil, the input of materials, suitable for pH increase, resulted in the restoration of biological activities and soil capacity to support vegetation. The association of chemical oxidation treatment and restoration techniques could present a good mean for soil rehabilitation. Finally, an index, which aimed to represent soil conditions based on the weight given to different parameters, was proposed and was used to evaluate changes induced by oxidation treatments and restoration techniques

Afin de contribuer à la préservation du patrimoine laitier libanais, l'objectif principal de cette étude a été de caractériser le fromage Darfiyeh, une variété traditionnelle fabriquée à partir de lait de chèvre cru et affinée dans une outre en peau de chèvre. Parallèlement à cet objectif, et pour mieux comprendre l'affinage du Darfiyeh, le transfert de composés d’arôme à travers la peau de chèvre a été étudié au moyen d’un dispositif expérimental original. Trois lots indépendants de production de Darfiyeh ont été analysés après 20, 40 et 60 jours d'affinage. Les résultats des analyses physico-chimiques ont montré que le Darfiyeh est un fromage à pâte mi-dure, caractérisé par une protéolyse et une lipolyse modérées. Le 3-méthyl butanal, le 1-phényl éthanol et le 1-octanol contribuent à la fraction aromatique particulière du Darfiyeh. Les analyses microbiologiques ont mis en évidence (Log cfu.g-1 fromage) des lactobacilles mésophiles (7,1-10,4), des bactéries lactiques (BL) thermophiles cocciformes (6,6-8,4) et des lactobacilles thermophiles (5,5-7,2). Afin d'explorer l'écosystème naturel du Darfiyeh, une combinaison d'approches classiques et moléculaires a été appliquée. L'identification classique a révélé des espèces appartenant aux genres Streptococcus, Enterococcus, Lactococcus et Lactobacillus. Les profils de Temporal Temperature Gel Electrophoresis (TTGE) ont révélé des bandes communes avec l'identification classique. Des analyses menées par PCR spécifiques de l'espèce bactérienne ont confirmé la présence of S. thermophilus, E. faecium, E. durans, Lc. lactis subsp. lactis et Lc. lactis subsp. cremoris. La PCR en temps réel a permis la quantification de S. thermophilus et d' E. faecium, avec un seuil de détection respectif de 104 ufc.g-1 de Darfiyeh et une gamme de 107-109 ufc.g-1 de Darfiyeh, respectivement. Pour le transfert d'arômes à travers la peau de chèvre, l'absorption et la perméabilité de trois molécules aromatiques modèles (2-butanone, 2,3-butanedione, 2-butanol) dans un système solution aqueuse/peau de chèvre a été suivi, après 20, 40 et 60 jours d'exposition. Jusqu'à 40 j d'exposition, le transfert des molécules aromatiques était régi par leurs propriétés physico-chimiques et par le temps d'exposition. Après 60 j, le système solution aqueuse/peau s’est traduit par une tendance à la migration des molécules modèles depuis la peau vers la solution simulée de Darfiyeh. Autrement dit, le transfert de ces molécules modèles dans le système simulé paraît s’établir de façon cyclique. Ce travail est une première contribution scientifique et technique au secteur de la valorisation durable de la tradition fromagère libanaise / In order to contribute for the preservation of the Lebanese dairy heritage, the aim of this study was to characterize Darfiyeh cheese, a traditional variety made from raw goats' milk and ripened in goat's skin. Parallel to this objective, and for a better understanding of Darfiyeh ripening, aroma transfer towards goat skin was conducted through an experimental device. Three independent batches of Darfiyeh production were analyzed after 20, 40 and 60 days of ripening. Physico-chemical results showed that Darfiyeh is a semi-hard goat's milk cheese which undergoes a moderate proteolysis and lipolysis. 3-methyl butanal, 1-phenyl ethanol and 1-octanol contribute to the distinctive aromatic fraction of Darfiyeh. As for microbiological results, bacterial counts (Log cfu.g-1 cheese) for mesophilic lactobacilli (7.1-10.4), thermophilic coccal-shaped lactic acid bacteria (LAB) (6.6-8.4) and thermophilic lactobacilli (5.5-7.2) were found. In order to explore Darfiyeh natural ecosystem, a combination of classical and molecular approaches was applied. Classical identification revealed members of the genera Streptococcus, Enterococcus, Lactococcus and Lactobacillus. Temporal Temperature Gel Electrophoresis (TTGE) profiles revealed common bands between the classical identification. Species-specific polymerase chain reactions (PCR) confirmed the presence of S. thermophilus, E. faecium, E. durans, Lc. lactis subsp. lactis and Lc. lactis subsp. cremoris. Real time PCR enabled quantification of S. thermophilus and E. faecium, with a detection threshold of 104 ufc.g-1 of Darfiyeh and a range of 107-109 ufc.g-1 of Darfiyeh, respectively. As for aroma transfer towards goat skin, absorption and permeability of three model aroma molecules (2-butanone, 2,3-butanedione, 2-butanol) in aqueous solution/goat skin system were followed, after 20, 40 and 60 days of exposure. Until 40 days, the transfer of aroma molecules was dependent on their physico-chemical properties and exposure time. After 60 days, the molecules migrate from the skin to the aqueous solution. Therefore, in such a simulated system, aromatic molecules seem to move under a cyclic way. This work is a first scientific and technical contribution to the preservation of traditional Lebanese cheesemaking

La thérapie génique est une forme de médecine moléculaire qui a des potentialités majeures dans le traitement d'un grand nombre de maladies héréditaires ou de cancers. Les acides nucléiques doivent pénétrer à l'intérieur des cellules et interagir avec la machinerie génétique présente. Une des plus grandes limitations dans l'application de ce type de traitement est le développement d'un vecteur sûr et efficace pour transporter ces molécules dans les cellules. Les vecteurs sont classifiés traditionnellement en deux grandes catégories : viral et non-viral. Les vecteurs de type viral sont les plus efficaces et utilisés dans la majorité des tests cliniques, cependant ils peuvent provoquer une forte réponse immunitaire et ils ont des coûts importants de production. Les vecteurs de type non viral peuvent être fabriqués facilement en grande quantité, ne sont pas immunogéniques et possèdent l'avantage de pouvoir être modulés en fonction des applications. Les lipides et les polymères cationiques sont les plus étudiés à cause de leurs propriétés de complexation avec les acides nucléiques. Cependant ces complexes chargés positivement peuvent être toxiques à cause de leur interaction non spécifique avec les membranes cellulaires, ce qui limite leur utilisation in vivo. Cela a conduit à la recherche de nouveaux vecteurs, neutres ou anioniques, pour diminuer les problèmes de cytotoxicité. Les nucléolipides (NL) sont des molécules hybrides bioinspirées amphiphiles formées d’une partie hydrophile nucléotidique et d’une partie hydrophobe lipidique. Ces molécules sont donc capables de s'autoassembler et de former des structures supramoléculaires avec des propriétés physico-chimiques principalement liées à la nature chimique de la tête polaire nucléotidique. L'interaction avec des acides nucléiques est alors possible grâce à des interactions de type Watson-Crick, cependant elles ne sont pas suffisamment élevées pour former un complexe stable pour être utilisées dans les conditions biologiques. La synthèse d’une nouvelle famille de molécules hybrides de type amino-nucléo-lipides (ANL) a été développée dans l'objectif d'améliorer les interactions et donc la complexation grâce à la présence d'un acide aminé sur la partie polaire de l’amphiphile. La synthèse chimique des ANLs avec différents acides aminés (glycine, phénylalanine) en position 5' du sucre et différentes lipides (dimiristoyl, dioleoyl) estérifié au phosphate présent en position 3' a été réalisée. Les études physico-chimiques réalisées par diffusion dynamique de la lumière (DLS), cryo-microscopie électronique (cryo-TEM) et diffusion des rayons X aux petits angles (SAXS) ont permis d'étudier la morphologie et l'organisation structurale des objets supramoléculaires formés par ces molécules. Ces études ont en particulier permis de mettre en évidence la relation entre la structure chimique et les propriétés physico-chimiques. Les NLs forment des vésicules unilamellaires et les ANLs, grâce à la présence de l'acide aminé sur la tête polaire, forment des vésicules de type multilamellaire. L'étude des interactions, réalisée par calorimétrie à titration isotherme (ITC), entre les vésicules et un acide nucléique modèle (poly A) montre qu'il existe une interaction entre les deux partenaires, mais que cette interaction n'est pas suffisamment forte pour envisager des applications biologiques. La stratégie d'utiliser un cation divalent, comme le calcium, pour ponter les charges négatives des nucléolipides et les charges négatives des acides nucléiques a été envisagé et les complexes ont été étudiés par SAXS. Enfin, des transitions de morphologies observées en fonction de la concentration ont été étudiées par DLS et cryo-TEM. Ces transitions de morphologies ont pu être corrélées à des effets de force ionique sur les propriétés de la tête polaire. / Gene therapy is a molecular medicine and a very powerful tool for the treatment of several diseases such as inherited disorder and cancer. Nucleic acids must penetrate into cells in order to interact with their genetic material. Currently the main limitation to the application of this treatment towards clinics is the lack of robust, safe and efficient gene delivery vectors. The two major classes of vectors are those based on recombinant viruses and those based on non-viral systems. Viral vectors are the most efficient and used in several clinical trials, however they can elicit a strong immune reaction and they possess high cost of production. Non-viral vectors are less immunogenic and can be easily produced on a large scale. A large variety of both cationic lipids and polymers have been developed due to their ability to interact spontaneously with negatively charged nucleic acids to form complexes. However these positively charged complexes can present some toxicity due their non-specific interaction with cell membranes and seric proteins. This is the main limitation for their clinical use. For this purpose, new vectors, neutral or anionic, have to be developed in order to diminish the cytotoxicity and increase the circulation time. Nucleotide-lipids (NLs) are bio-inspired amphiphilic hybrid molecules composed of a hydrophilic nucleotidic moiety and a hydrophobic lipophilic moiety. These molecules are able to self-assembly to form supramolecular structures which possess particular physico-chemical properties due to the chemistry of their polar head. These molecules can interact with a nucleic acid by Watson-Crick base pair interactions, however they are not sufficiently strong to form a stable complex that can be used for a biological application. A new chemical family of hybrid amphiphile, amino acid-nucleotide-lipids (ANLs), has been developed in order to increase the interactions and the stability of the complex thank to the presence of the amino acid on the polar head. Herein, we have synthesized novel amino acid-nucleotide-lipids, presenting phenylalanine (or glycine) and thymidine residues and saturated (dimiristoyl) or unsaturated (dioleoyl) diacyl glycerol lipid. The morphology and the structural organization of the supramolecular objects formed by these molecules was studied by dynamic light scattering (DLS), cryo-electron microscopy (cryo-TEM) and small angle X-ray scattering (SAXS). These studies allowed investigating the relation between the chemical structure and the physic-chemical properties. The amino acids, inserted at the 5′ position of the nucleotide-lipids, stabilize multilamellar systems, whereas unilamellar vesicles are formed preferentially in the case of nucleotide-lipids. Both NLs and ANLs exhibit weak interactions with complementary polyA RNA as revealed by isothermal titration calorimetry (ITC) investigations, however they are not sufficiently strong to form a stable complex that can be used for a biological application. The use of multivalent cations, such as Ca2+, which bridge the phosphate groups on the lipid polar heads with those of the backbone of nucleic acids, to form ternary complexes, has been investigated by SAXS. Finally, a structural study, by DLS and cryo-TEM of NLs aggregates in aqueous solutions as a function of ionic strength and surfactant concentration, has been conducted in order to investigate the different morphologies of the systems.

Les liquides ioniques sont des produits chimiques avancés promis selon de nombreuses personnes à un brillant avenir dans un certain nombre d’applications stratégiques qui pourraient servir un avenir plus vert dans la chimie et les développements technologiques liés à l'énergie. Cependant, le manque de données thermodynamiques appropriées ainsi que les discours réducteurs portant sur une haute stabilité thermique et l’absence de tout danger intrinsèque comme tendances absolues de ces produits chimiques ont conduit à entreprendre ce travail de thèse avec comme objectif de développer une méthodologie dédiée et pertinente visant à caractériser les profils de dangers physico-chimiques de ces molécules chimiques. En effet, étant donné le nombre potentiellement élevé de liquides ioniques, une meilleure stratégie consistant à explorer les propriétés réelles de ces matériaux, sans négliger les risques physico-chimiques potentiels, y compris les risques d’incendie, constitue une contribution de taille à leur utilisation verte. A cet effet, des outils expérimentaux ont été mis en œuvre et utilisés, les résultats obtenus confirment jusqu’ici que le potentiel de dangers physico-chimiques doit être évalué au cas par cas.Finalement, les perspectives concernant les travaux futurs en ce qui concerne une meilleure connaissance et une appréciation des questionnements liés aux dangers physico-chimiques sont donnés dans le but de servir une conception et une utilisation durable des liquides ioniques. / Ionic liquids are advanced chemicals promised according to many people to a brilliant future in a number of strategic applications that might serve greener future in chemistry and energy related technological developments. However, lack of appropriate thermodynamic data as well as some what misleading discourse advocating on high thermal stability and absence ofany intrinsic hazard as absolute trends of these chemicals have led to initiate this thesis workwith the objective to develop a dedicated and pertinent methodology aiming at characterizing the comprehensive physico-chemical hazards profile of such chemical.Indeed, given the huge potential number of ionic liquids, a better strategy consists in exploring real world properties of these materials, not neglecting potential physico-chemical hazards,including the fire hazards, as to contribute to their green use. For that purpose, experimental tools have been implemented and used; obtained results so far confirm that the physico-chemical hazards potential must be assessed on a case by case approach. Finally, perspectives regarding future work with regard to better knowledge and assessmentof ionic liquids physico-chemical related issues are given with the aim of serving sustainable design and use of ionic liquids.

La libération des composés d'arôme détermine la qualité aromatique des produits alimentaires, et contribue ainsi aux choix et préférences des consommateurs. Dans ce contexte, la compréhension et la modélisation de la cinétique de libération est un défi scientifique et un enjeu de santé afin de pouvoir formuler des produits en intégrant des critères nutritionnels et sensoriels. Ce travail de thèse a permis d'étudier et de modéliser les mécanismes en bouche responsables de la dynamique de libération des stimuli olfactifs lors de consommation d'un aliment liquide ou solide chez l'homme.<br />• Dans un premier temps, un modèle mécanistique décrivant la libération des composés d'arôme au cours de la consommation d'un aliment liquide ou semi-liquide a été développé. Ces produits ont un temps de résidence en bouche très court et ne nécessitent pas de manipulation intra-orale complexe. Le modèle a été construit sur la base de bilans de matière prenant en compte des mécanismes de transfert entre certains sous-compartiments du système, ainsi que les conditions spécifiques aux différentes étapes de la consommation. Une comparaison du modèle avec des données de libération in vivo lors de la consommation de fluides newtnoniens aromatisés avec du diacétyle et de l'hexanoate d'éthyle a été effectuée. Cette étude nous a permis de comprendre le rôle du résidu post-pharyngé et de la viscosité sur la libération des composés d'arôme : • l'épaisseur du bol tapissant les muqueuses a pu être estimée à environ 15µm; • contrairement à l'hypothèse initiale, il a été mis en évidence que les propriétés pertinentes à prendre en considération pour la libération des composés d'arôme à partir d'un fluide newtonien sont celles d'un mélange de produit hautement dilué par la salive. • Dans un second temps, le modèle a été adapté à des produits nécessitant une mastication. Pour en rendre compte, de nouveaux mécanismes ont été intégrés: phénomènes de transfert de matière et de dissolution du produit dans la salive, génération d'une surface d'échange produit/fraction liquide du bol et l'ouverture vélopharyngienne lors de la mastication du produit. Le modèle a ensuite été confronté avec les données de libération du propanoate d'éthyle in vivo lors de la consommation de matrices fromagères modèles. Le modèle a pu être ajusté de façon satisfaisante à l'ensemble des données expérimentales et les deux paramètres inconnus de notre modèle (la vitesse d'incorporation moyenne de salive dans le bol au cours de la consommation et la fréquence d'ouverture du vélopharynx) ont pu être estimés. Cette étude nous a permis de comprendre le rôle de la mastication sur la libération des composés d'arôme lors de la consommation d'aliments solides. De plus, l'étude de la libération de la 2-nonanone a permis de mettre en évidence un phénomène d'adsorption sur les muqueuses pour cette molécule. • Enfin, il ressort de la comparaison des deux modèles que les paramèters clés gouvernant la libération des composés d'arôme ne sont pas les mêmes selon la catégorie de produit (liquide ou solide) considérée: • lors de la consommation d'aliments liquides ou semi-liquides, le coefficient de transfert de matière dans le bol, la fréquence respiratoire de l'individu et l'épaisseur du résidu post-pharyngé sont les trois facteurs clés gouvernant la libération des composés d'arôme; • en revanche, lors de la consommation de produits solides mastiqués, ce sont la vitesse d'incorporation moyenne de salive dans le bol, la fréquence d'ouverture du vélopharynx et la durée de mastication qui sont les trois paramètres ayant un effet majeur sur les cinétiques de libération. La démarche de modélisation nous a permis de mieux comprendre les parts relatives du produit, de l'individu, et de l'interaction produit-individu sur la libération des composés d'arôme au cours de la consommation d'un aliment. / Delivery of aroma compounds to olfactory receptors determines the aromatic quality of food products and contributes to consumer choices and preferences. Therefore, understanding and modelling the release kinetic is a scientific challenge and a health issue in order to formulate products of both high nutritional and sensory quality. This thesis studied in-mouth mechanisms responsible of the dynamics of olfactory stimuli release during food consumption. • First, a mechanistic model describing the aroma compounds release during consumption of a liquid or semi-liquid food has been developed. These products have a very short in-mouth residence time and do not require complex intra-oral manipulation. The model takes into account mass balances, transfer mechanisms occurring between some sub-compartments of the system, and the specific conditions at the different stages of consumption. A comparison of the model predictions with in vivo release data during the consumption of Newtnonien fluids flavored with diacetyl and ethyl hexanoate was performed. This study highlighted the role of post-pharyngeal residue and viscosity on the aroma compounds release: • the thickness of bolus covering the mucous membranes has been estimated at about 15μm; • it was found that the relevant properties to be considered for the release of aroma compounds from a Newtonian fluid are those of a mixture highly diluted by saliva. • Second, the model previously developed was adapted for products requiring chewing. It takes into account the phenomena of mass transfer and dissolution of the product in the saliva during chewing. The generation of a product/liquid contact surface as well as the velopharyngial opening that occurs during the mastication of the product were also integrated into the model. The model was then confronted with in vivo release data for ethyl propanoate during consumption of four cheese matrices. All simulations have been satisfactorily fitted to experimental data and the two unknown parameters of our model (the average rate of saliva incorporation into the bolus and the frequency of velopharyngial opening) could be estimated. This study has enabled us to understand the role of mastication on the release of aroma compounds during consumption of solid food: • the opening of velopharynx during intra-oral manipulation of the product produces a continuous supply of aroma compounds in the nose; • the residence time of solid product in the mouth are much longer than for the consumption of liquid and semi-liquid foods, allowing the secretion of significant volumes of saliva. In addition, the study of the release of 2-nonanone highlighted an adsorption phenomenon on the mucous membranes for this molecule. • Finally, sensitivity analysis of the two release models indicates that: • when eating a liquid or semi-liquid food, the mass transfer coefficient in the bolus, the breath rate and the thickness of post-pharyngeal residue are the three key factors governing the release of aroma compounds; • however, when eating a solid food product, it is the average rate of saliva incorporation into the bolus during consumption, the frequency and duration of velopharyngeal opening, and the mastication time which are the three parameters that have major effects on the kinetics of release. The modeling approach allowed us to better understand the relative effects of the product, the individual, and individual-product interaction on the release of aroma compounds during food consumption. The results of this work indicated that the most important parameters depend on the category of product (liquid or solid) under consideration.

Surface water is globally becoming more and more a scarce resource, and in Addis Ababa the capital of Ethiopia, river water quality has been degraded due to anthropological forcing for many years. Therefore, the study objective was to investigate causes and impact of surface water pollution in Kebena and Great Akaki rivers. The technical aspect of the study focused on analysing the parameters E. coli, phosphate, nitrate and total ammonia nitrogen in 34 different sampling sites in the western part of the Great Akaki catchment. The other aspect was to evaluate authorities’ and companies’ perspective on the water quality, usage and future plans to mitigate further pollution of rivers. Another perspective was to interview households and farmers regarding their view on usage, water quality and health risks. The main finding was a high surface water contamination in both Kebena and Akaki river, throughout the city, mostly from domestic, municipality and industrial wastewater and solid waste. E. coli concentrations exceeded thresholds given by WHO. Concentrations of phosphate and total ammonia nitrogen strongly indicated eutrophication. Nitrate values were lower than expected with no perceived healthrisk. The interview study with authorities, households and farmers indicated irrigation as the main usage. Little to moderate health risks perceived by farm users and high health risks perceived by authorities for farmers were found. Therefore, addressing a stronger collaboration between authorities and the local community is important. In addition, the implementation of mitigation strategies should be strengthened and the stakeholders need to be accountable for their actions. A continued monitoring of pollutants as well as a multi-sectoral approach to solidwaste and wastewater management will help improve the river water quality. / Ytvatten blir globalt allt mer en knapp resurs och i Addis Abeba, huvudstaden i Etiopien, har flodernas vattenkvalitet under många år försämrats på grund av antropogen påverkan. Denna studie syftar till att undersöka orsaker och påverkan på ytvattenföroreningar i floderna Kebena och Akaki. Den tekniska aspekten av studien inkluderar vattenanalyser av parametrarna E.coli, fosfat, nitrat och totalt ammonium kväve som utfördes på 34 olika provtagningsplatser i västra delen av Great Akakis avrinningsområde. Den andra aspekten var att utvärdera myndigheters och företags perspektiv på vattenkvalité, flodvattnets användningsområden och framtida planer för förbättring av föroreningsgraden i floderna. Ett annat perspektiv var att intervjua hushåll och lantbrukare angående deras bild av ytvattenanvändning, om vattenkvaliteten och hälsorisker. Studiens huvudsakliga upptäckt är en genomgående hög föroreningsgrad i stadens flodvatten. Föroreningen består till största del av avlopp och avfall från hushåll, kommuner och industrier. Koncentrationerna av E.coli överskred WHO:s gränsvärden. Halterna av fosfat och totalt ammoniumkväve indikerade övergödning. Nitratvärdena visade lägre halter än förväntat och därmed ingen påvisad hälsorisk. Intervjustudien med myndigheter, hushåll och lantbrukare påvisade att ytvattnet mest används för bevattning av åkermark. Lantbrukarna uppfattade en liten till medelhög hälsorisk med denna användning, medan myndigheter ansåg att lantbrukarna utsattes för en hög risk. Därför är ett starkare samarbete mellan myndigheter och samhället viktigt. Dessutom behöver implementationen av förbättringsåtgärder förbättras och alla aktörer måste göras ansvariga för sina handlingar. En fortsatt övervakning av föroreningar och ett multi-disciplinärt arbetssätt vid avfall- och avloppshantering kommer att vara till hjälp vid förbättring av vattenkvaliteten i floden.

Para avaliar a influência do processo de secagem da glândula mamária sobre as características físico-químicas e celulares da secreção láctea foram avaliadas 216 amostras provenientes de 27 glândulas mamárias de 15 cabras da raça Saanen consideradas sadias, sendo que essas mamas foram separadas de acordo com o resultado do exame microbiológico da secreção láctea, caracterizando dois grupos: com e sem isolamento bacteriano, composto por 17 e 10 mamas, respectivamente. A partir do dia da interrupção da ordenha e em mais sete momentos (1, 3, 5, 7, 10, 15 e 30 dias após o início da secagem) foi feita a inspeção e a palpação das glândulas mamárias e posterior avaliação do aspecto da secreção láctea. Imediatamente após o exame clínico foram colhidas amostras de leite para a realização do exame microbiológico e das determinações das características físico-químicas e celulares (pH, eletrocondutividade, cloreto, lactose, proteínas, gordura, sólidos totais, índice cloreto/lactose, California Mastitis Test (CMT) e contagem de células somáticas). As glândulas mamárias apresentaram sua maior distensão no terceiro dia após a interrupção da ordenha, mesmo período em que foi possível observar o aparecimento de edema na base dos tetos e em toda a glândula, desaparecendo após o quinto dia, quando teve início a involução do úbere e a freqüência de mamas distendidas começou a declinar, sendo que ao final do processo 37,04% das mamas observadas eram classificadas como não distendidas. A secreção láctea inicial com aspecto de leite foi substituída a princípio por uma de coloração creme clara e mais tardiamente por uma de aspecto aquoso, sendo que ao final do processo as duas últimas eram mais observadas que a com aspecto de leite. Apenas o pH, a eletrocondutividade, os teores de cloretos, lactose, proteínas e índice cloreto/lactose sofreram uma significativa influência do processo de secagem, sendo que os valores do pH, eletrocondutividade, cloretos, proteínas e índice cloreto/lactose aumentaram e os valores de lactose declinaram. O CMT também sofreu uma influência do processo de secagem, mais especificamente as respostas \"traços\" e \"duas cruzes\". Estas, de maneira geral, aumentaram no decorrer da secagem, sendo que no último momento de colheita ocorreu uma queda na freqüência da resposta \"traços\" e a freqüência da resposta \"duas cruzes\" teve uma pequena diminuição no meio do processo. Ainda foi observado que, apesar de existirem diferenças estatísticas significativas entre valores das características físico-químicas e celulares das amostras de secreção láctea com e sem isolamento bacteriano, as diferenças são isoladas e limitadas a algumas características estudadas, impossibilitando afirmar que o processo de secagem dos dois grupos difere de maneira significativa. / To evaluate the influence of the mammary gland drying off on the physiochemical and cellular characteristics in the milk secretion they were appraised 216 coming samples of 27 mammary glands of 15 Saanen goats considered healthy, those mammas were separate in agreement with the result of the microbiological test of the milky secretion characterizing two groups: one with 17 mammas without bacterial isolation and other with 10 mammas that presented positive bacterial isolation. Starting from the day of the interruption of the milking and in more seven moments (1, 3, 5, 7, 10, 15 and 30 days after the beginning of the drying off) it was made the inspection and palpation of the mammary glands with subsequent evaluation of the aspect of the milky secretion. Immediately after the exam they were picked samples of milk for the accomplishment of the microbiological test and of the determinations of the physiochemical and cellular characteristics (pH, electrical conductance, chloride, lactose, proteins, fat, total solids, chloride/lactose ratio, California Mastitis Test (CMT) and somatic cell count). The mammary glands presented its largest distention in the third day after the milking interruption, same period in that was possible to observe the edema emergence in the teats base and in the whole gland, that disappeared after the fifth day, when involution of the udder begins and the frequency of distended mammas began to refuse, and at the end of the process 37,04% of the observed mammas they were classified as no distended. The initial milky secretion with aspect of milk was substituted at first for one of coloration cream clear and, so late, for one of aqueous aspect, and in the end of the process those two last they were more observed that the secretion with aspect of milk. Just the pH, electrical conductance, chloride, lactose, proteins and chloride/lactose ratio suffered a significant influence of the drying off, and the values of the pH, electrical conductance, chloride, proteins and chloride/lactose ratio increased and the lactose values refused. CMT also suffered an influence of the drying off, more specifically the answers \"lines\" and \"two crosses.\" These, in a general way, increased in elapsing of the drying off, and in the last moment studied it happened a fall in the frequency of the answer \"lines\" and the frequency of the answer \"two crosses\" had a small decrease in the middle of the process. It was still observed that in spite of significant statistical differences exist among values of the physiochemical and cellular characteristics of the samples of milky secretion with isolation and without bacterial isolation, the differences are punctual and limited to some studied characteristics, disabling to affirm that the process of drying of the two groups differs in a significant way.

As propriedades no estado sólido de um insumo farmacêutico ativo (IFA) estão diretamente relacionadas com a eficácia terapêutica de um medicamento. Entre todas as propriedades físicas e químicas, que são dependentes doarranjo cristalinodo IFA, destaca-se a solubilidade e a estabilidade, que são as características mais importantes de um fármaco, na medida em que podem alterar a biodisponibilidade. Dentro dessa perspectiva, a engenharia de cristaistem cada vez mais se consolidado como uma estratégia para aperfeiçoar as propriedades no estado sólido de um IFA. Neste trabalho, oito novas formas sólidas cristalinas, entre sais e cocristais, dos fármacos com ação anti-tuberculose etionamida (ETH), etambutol (ETB) e isoniazida (INH), foram preparadas e tiveram suas estruturas cristalinas determinadas por difração de raios X por monocristal. Além disso, todas as modificações cristalinas obtidas foram caracterizadas por difração de raios X por policristais, espectroscopia vibracional no infravermelho e Raman, calorimetria exploratória diferencial, termogravimetria e microscopia ótica térmica. Por fim, estudos de solubilidade em equilíbrio e de estabilidade em atmosfera úmida também foram realizados. A nova modificação cristalina da ETH, o sal nitrato de ETH, foi planejada visando melhorar a solubilidade aquosa deste IFA (&#8764;0,4 mg mL-1). Este é o primeiro relato de uma estrutura de ETH que cristalizou com quatro independentes pares iônicos (ETH+/NO3-) na unidade assimétrica. A análise do empacotamento cristalino e das interações intermoleculares permitiu estabelecer os atributos estruturais que explicam a menor estabilidade térmica e a elevada solubilidade (cerca de 300 vezes maior) deste sal, em comparação com a forma neutra da ETH. A preparação dos novos sais de ETB foi feito considerando o cárater básico da molécula, o que permitiu a obtenção de três novos sais a partir da reação do ETB com os ácidos oxalico, maleico e tereftálico. O desenho dessas formas sólidas teve como principal objetivo minimizar os efeitos da elevada higroscopicidade do ETB, que é tão prejudicial para formulações farmacêuticas. Este objetivo foi alcançado, uma vez que o sal oxalato de ETB, reportado neste trabalho, mostrou ser não-higroscópico. Por fim, quatro novos cocristais da INH, sendo dois cocristais polimórficos, foram preparados por meio de reações estequiométricas deste IFA com ácidos carboxílicos aromáticos derivados do ácido benzóico. O desenho dessas novas modificações cristalinas teve como intuito principal explorar a diversidade de síntons supramoleculares formados entre a INH e moléculas contendo o grupo COOH. A análise dos empacotamentos cristalinos das formas I e II do cocristal INH-PABA, mostrou que este é um típico caso de polimorfismo orientacional. Portanto, os resultados desta dissertação trazem importantes contribuições científicas para a diversidade de formas sólidas, apresentando revelância para o setor farmacêutico no que se refere ao desenvolvimento de novas formulações contendo fármacos anti-tuberculose. / Solid state properties of an active pharmaceutical ingredients (API) are directly related to drug performance. Among all physical and chemical properties dependent on the crystal arrangement of a drug, stability and solubility are the main ones that alter its bioavailability. In this sense, crystal engineering is a strategy to improve solid state properties of drugs. In this work, eigth new multicomponent solid forms (salts and cocrystals) of the tuberculostatic drugs ethionamide (ETH), ethambutol (ETB) and isoniazid (INH) were prepared and their crystal structures were elucidated by single crystal X-ray diffraction. In addition, the crystal modifications were also characterized by powder X-ray diffraction, infrared and Raman vibrational spectroscopy, differential scanning calorimetry, thermogravimetry and Hot-stage microscopy. Finally, equilibrium solubility studies and stability experiments in humid atmosphere were also performed. The new supramolecular modification of ETH, ethionamide nitrate salt, was designed in order to improve the aqueous solubility of the API (&#8764;0.4 mg ml-1). This salt is the first ETH structure that has been crystallized with four independent ionic pairs (ETH+/NO3-) in the asymmetric unit. The analysis of crystal packing and intermolecular interactions allowed to establish the structural attributes that explain the lower thermal stability and high solubility (about 300 times higher) of this salt compared to the neutral ETH form. Due its high hygroscopicity, ETB is known for catalyzing the degradation of isoniazid and rifampicinin pharmaceutical formulations. In order to avoid or even minimize these mutual drug-drug interactions, in this work we have focused on the development of less hygroscopic multicomponent solid forms of ETB. The preparation of the new ETB crystal modifications was carried out considering the pKa values of the molecule, which allowed the design of three new salts (namely oxalate, maleate and terephthalate). These salts were obtained from the reaction of ETB with the carboxylic acids: oxalic, maleic and terephthalic. The hygroscopic nature of these salts was evaluated and all of them were found to be hygroscopic, with exception of ETB oxalate. Finally, four novel INH cocrystals, being two polymorphic forms, with aromatic carboxylic acids derivatives were rationally prepared. The design of these crystal forms aimed to explore the diversity of supramolecular synthons formed between INH and molecules containing COOH. The crystal packing analysis of the INH-PABA cocrystals (polymorphs I and II), showed that this is a typical case of orientational polymorphism. Therefore, the scientific contributions of this work show the diversity of the solid forms and define candidates to new anti-tuberculosis API\'s solid formulations.

O arroz (Oryza sativa L.) é um dos cereais mais produzidos e consumidos no mundo, caracterizando como principal alimento para mais da metade da população mundial. Sua importância é destacada principalmente em países em desenvolvimento, tais como o Brasil, desempenhando papel estratégico em níveis econômico e social. Atualmente, é crescente o número de pesquisas que integram desde o melhoramento vegetal e o manejo da adubação até o aproveitamento dos nutrientes pelo organismo humano. O melhoramento de plantas pode ajudar a melhorar a dieta humana, com o desenvolvimento de plantas com maiores teores de vitaminas e micronutrientes. A introdução de produtos agrícolas biofortificados como o arroz, complementará as intervenções em nutrição existentes. Sendo assim, tornase imprescindível conhecer como as condições de cultivo influenciam na qualidade do grão e na importância para obtenção de um alimento com maior valor biológico e nutricional e com menores concentrações de antinutricionais. O objetivo deste trabalho foi analisar as características físico-químicas e nutricionais do arroz (Oryza sativa L.) integral, promovidas por diferentes adubações (experimento I) e aos grãos biofortificados (experimento II). Foram realizadas análises de composição centesimal, composição mineral, diálise in vitro de minerais, vitaminas do complexo B, conteúdo de compostos fenólicos e fitatos em grão, além das análises de parâmetros de qualidade em arroz como conteúdo de amilose aparente, propriedade de pasta e análise instrumental de cor. Os resultados obtidos no estudo assemelham-se aos citados nas literaturas quanto á composição centesimal e mineral, enfatizando a influência da cocção na composição nutricional do grão e em seus compostos como fenólicos e fitatos. Para o dialisado de cálcio e zinco no arroz biofortificado, a variação nos valores foi em média de 1,14% a 3,26% para cálcio e 0,80% a 3,43% para zinco. No experimento II, as perdas de vitaminas para o arroz integral biofortificado após o cozimento foi em torno de 33,4%, 52,17% e 76% para vitamina B1, vitamina B2 e vitamina B6, respectivamente. Por fim, conclui-se que a produção de variedades melhoradas seja através de adubações ou da escolha de melhores cultivares, que apresentam maior teor de minerais e vitaminas, foram eficientes para a melhora da disponibilidade nutricional dos grãos. / Rice (Oryza sativa L.) is one of most cereals produced and consumed in the world, characterized as the primary food for more than half the world population. Its importance is highlighted especially in developing countries such as Brazil, playing a strategic role in economic and social levels. There is increasing the number of studies since that integrate plant breeding and fertilizer management to use the nutrients the human body. Plant breeding can help improve the human diet, with the development of plants with higher levels of vitamins and micronutrients. The introduction of biofortified crops like rice, will complement existing nutrition interventions. Thus, it becomes essential to know as the cultivation conditions influence the quality of grain and the importance to obtain a food with higher nutritional and biological value and with lower concentrations of antinutritional. Threfore, the aim of this study was to analyze the pshycochemical and nutritional properties of rice (Oryza sativa L.) in full, caused by different fertilization (experiment I) and subjected to biofortification (experiment II). Were performed to analyze chemical composition, mineral, dialysis in vitro of minerals, vitamins B, content of phenolic compounds and phytates in grain, in addition to the analysis of quality parameters in rice as apparent amylase content, folder property and instrumental analysis of color. The results obtained in this study are similar to those cited in other literature as to the mineral and chemical composition, emphasizing the influence of cooking on the nutritional composition of grain and as phenolic compounds and phytates. For the dialysate calcium and zinc in rice biofortified crops, the variation in the values averaged 1.14% to 3.26% for calcium and 0.80% to 3.43% for zinc. Losses of vitamins to the rice after cooking was around 33.4%, 52.17% and 76% for vitamin B1, vitamin B2 and vitamin B6, respectively. Finally, we conclude that the production of improved varieties is through fertilization or the best choice of cultivars, that have a higher content of minerals and vitamins, were effective to improve the availability of nutrients of the grains.