La fonctionnalisation C(sp3)-H catalysée par des métaux de transitions, ouvre de nombreuses perspectives en synthèse organique, permettant des voies d'accès plus économes en atomes, et en étapes à des molécules à forte valeur ajoutée. Dans cette optique, une méthode efficace permettant l'arylation des liaisons C(sp3)-H en position α d'un groupement attracteur, plus communément appelée α -arylation a récemment fait l'objet d'une attention toute particulière de la part de la communauté scientifique. Le travail détaillé dans ce manuscrit décrit les dernières avancées de cette méthodologie, ainsi qu'une variante «β-arylation » développée au laboratoire qui constitue une évolution significative dans le domaine de l'arylation regiosélective des liaisons C(sp3)-H non activées. Dans le cadre de ce projet de thèse nous nous sommes efforcés de développer cette nouvelle réaction que nous avons pu optimiser pour l'étendre à une famille plus étendue de substrats de type amino-esters. Dans la continuité de ce travail nous avons réalisé la première réaction d'arylation migratoire sélective d'amino-esters pouvant aller jusqu'à la position η d'une chaîne alkyle linéaire. Enfin, dans le but d'accéder à de nouvelles molécules à plus haute valeur ajoutée, nous avons pu appliquer notre méthodologie aux acetals de cétènes silylés permettant de dépasser certaines limitations du système existant. Dans ce cas précis, des conditions plus douces (sans base forte) ont permis l'arylation de substrats dits sensibles et par extension la synthèse de lactones fonctionnalisées / The transition metal catalysed functionalization of C(sp3)-H bonds unlocks numerous perspectives within organic synthesis in terms of atom economical access routes to otherwise difficult to synthesise molecules. One efficient method to exact such transformations involves the exploitation of an activated C-H bond situated adjacent to an activating electron withdrawing group, allowing facile insertion of a transition metal catalyst species and subsequent functionalization with a new species (normally an aryl group). This strategy is generally termed ‘α-functionalization’. The work detailed within this manuscript describes a diversion from the classic, and well documented α-functionalization reaction, in which rearrangement steps within the catalytic cycle give rise to β- and more remote substrate functionalization. The first new methodology to be described involves a fundamental extension to the in-house developed β-arylation reaction, in which, through careful substrate and ligand choice, this methodology could be applied to achieve the functionalization of simple ester enolates in remote γ- to η - positions. The developed strategy allowed the synthesis of a small range of interesting homophenylalanine analogues, and higher homologues. The second methodology to be described involves a necessary modified protocol for the β-arylation reaction, in which silyl ketene acetals were exploited as mild metal-enolate surrogates, allowing the coupling of base-sensitive substrates. The previously described reaction scope has been extended in terms of both the electrophile and nucleophile coupling partners through the development of mild reaction conditions, which subsequently allowed application of several products towards the synthesis of lactones

Dans le domaine de la santé, l’analyse de l'haleine expirée offre un outil simple et non invasif pour le diagnostic précoce des maladies. Les capteurs de gaz à base de SnO2 modifies semblent être des dispositifs prometteurs pour détecter les gaz polaires tels que l'ammoniac. Dans cette étude, la fonctionnalisation de la surface de SnO2 a été réalisée afin d'obtenir un capteur de gaz sensible et sélectif à l'ammoniac, qui fonctionne à température ambiante. La première étape de la fonctionnalisation est la fixation covalente d’un film de 3-aminopropyltrethoxysalane (APTES) sur SnO2 en phase vapeur ou liquide. Les caractérisations effectuées par Spectroscopie Infrarouge et Spectrométrie photoélectronique X montrent qu’une quantité plus importante d'APTES a été greffée en phase liquide hydratée. La deuxième étape consiste à fonctionnaliser le SnO2-APTES avec des molécules contenant du chlorure d'acyle avec différents groupes tel que des groupes alkyle, acide et ester. Les capteurs modifiés par des acides et des esters sont sensibles à l’ammoniac entre 0,2 et 10 ppm à température ambiante. Cependant, le SnO2 APTES modifié par l’ester s'est révélé être plus sélectif que le capteur modifié par l'acide pour l’ethanol et le mondxyde de carbone. Ces résultats impliquent que la réponse est générée par les groupes fonctionnels acide et ester, NH3 modifie le moment dipolaire de la couche moléculaire greffée, ce qui entraine une modification de la conductance de SnO2. Le fonctionnement à température ambiante est l'un des avantages de ces capteurs, tout comme leur sélectivité à l'ammoniac en regard d'autres gaz tels que l'éthanol, le monoxyde de carbone et l'acétone. / One of the major challenges in the modern era is how we can detect the disease when we are still feeling healthy via noninvasive methods. Exhaled breath analysis is offering a simple and noninvasive tool for early diagnosis of diseases. Molecularly modified SnO2 sensors seem to be promising devices for sensing polar gases such as ammonia. SnO2 surface functionalization was performed in order to obtain sensitive and selective ammonia gas sensor that operates at room temperature. The first step of functionalization is the covalently attachment of 3-aminopropyltriethoxysilane (APTES) film on SnO2 in vapor or liquid phases. The characterization performed by the Infrared Spectroscopy and X-ray Photoelectron Spectroscopy, show that more APTES were grafted by hydrous liquid phase silanization. The second step was the functionalization of APTES modified SnO2 with molecules having acyl chloride of end functional groups molecules such as alkyl, acid and ester groups. Pure SnO2 and APTES modified SnO2 sensors did not show any significant sensitivity to ammonia (0.2-100 ppm) at 25 °C. On the contrary, acid and ester modified sensors are sensitive to ammonia between 0.2 and 10 ppm at room temperature. However, ester modified SnO2 was more selective than acid modified sensor regarding the ethanol and carbon monoxide gases. Ammonia variates the attached molecular layer’s dipole moment which leads to change in SnO2 conductance. Working at ambient temperature is also one of the advantages of these sensors in addition to the selectivity to ammonia regarding other gases such as ethanol, carbon monoxide and acetone.

Ce travail porte sur la caractérisation physico-chimique de l'huile de Jatropha Curcas et sa capacité à remplacer l'huile minérale dans les transformateurs de puissance. Ce produit présente plusieurs avantages sur les autres huiles végétales comme l'huile de palme ou l'huile de colza, qui recommandent sa production et son utilisation. En effet, la plante de Jatropha Curcas peut être cultivée sur des sols pauvres à faibles précipitations, évitant ainsi d'utiliser des sols plus fertiles pour sa culture permettant ainsi aux petits exploitants de réserver leurs terres aux cultures de base. Cette plante peut pousser facilement dans des zones où les niveaux de précipitations annuelles sont nettement inférieures à celles requises par d'autres espèces telles que le colza, le tournesol, le soja, le maïs, le palmier à huile et d'autres. Elle peut être cultivée sur tous les types de sol en Indonésie, même sur des terres arides, dans de nombreuses régions de l'Indonésie orientale, inexploitées en raison des difficultés à planter d'autres cultures. En outre, l'huile de Jatropha Curcas est un produit non alimentaire. En faisant subir à l’huile de Jatropha Curcas brute une estérification à base alcaline avec de l'hydroxyde de potassium (KOH), on obtient de l’huile de méthylester de Jatropha Curcas (JMEO) dont la viscosité et l’acidité sont acceptables pour les équipements à haute tension en particulier pour les transformateurs de puissance. Les propriétés physico-chimiques et électriques de JMEO ont été mesurées ainsi que celles de l'huile minérale (MO) pour la comparaison. Pour les propriétés physico-chimiques, il s’agit de la densité relative, la teneur en eau, la viscosité, l'acidité, l'indice d'iode, la corrosivité, le point d'éclair, le point d'écoulement, la couleur, l'examen visuel, et la teneur en ester méthylique. Quant aux propriétés électriques, elles concernent la rigidité diélectrique sous différentes formes de tension (alternative, continu et choc de foudre), les phénomènes de pré-claquage et de claquage sous choc de foudre, les décharges glissantes sur les surfaces de carton comprimé, immergé dans JMEO et MO. Les résultats obtenus montrent que les tensions de claquage moyennes en continu et en choc de foudre des huiles JMEO et MO sont très proches ; la tension de claquage moyenne de JMEO est même plus élevée que celle de l'huile minérale (de type naphténique). La mesure des tensions de claquage des mélanges d'huiles «80% JMEO + 20% MO» et «50% JMEO et 50% MO» montrent que la tension de claquage du mélange «80% JMEO + 20% MO» est toujours supérieure à celle de l'huile minérale sous tensions alternative et continue. Cela indique que le mélange d'huile minérale et de JMEO avec un rapport de 20:80 ne dégrade pas ses performances. Le mélange d'huiles peut se produire lors du remplacement de l'huile minérale par JMEO dans les transformateurs installés et en exploitation. L'analyse des caractéristiques des streamers (la forme, le temps d'arrêt, le courant associé et la charge électrique) se développant dans les huiles JMEO et MO sous tension impulsionnelle de foudre, montre une grande similitude. Aussi, la longueur finale (Lf) et la densité des branches des décharges surfaciques se propageant sur le carton comprimé immergé dans l'huile de Jatropha Curcas de méthylester (JMEO) et de l'huile minérale (MO), sous tensions de choc de foudre positif et négatif (1,2/50 μs), pour deux configurations d'électrodes divergentes (électrode pointe haute tension perpendiculaire et tangente au carton, respectivement), sont fortement influencées par l'épaisseur du carton comprimé. Pour une épaisseur donnée, Lf augmente avec la tension et décroît lorsque l'épaisseur augmente. Lf est plus long lorsque la pointe est positive que lorsque la pointe est négative. Pour une tension et une épaisseur du carton comprimé donnée, les valeurs de Lf dans l’huile minérale et l’huile JMEO sont très proches. [...] / This work is aimed at the investigation of the physicochemical characterization of Jatropha Curcas seeds oil and its capacity to be an alternative option to replace mineral oil in power transformers. This product presents several advantages that recommend both its production and usage over those of other vegetable oils as crude palm oil and rapeseeds oil. Indeed, it may be grown on marginal or degraded soils avoiding thus the need to utilize those more fertile soils currently being used by smallholders to grow their staple crops; and it will readily grow in areas where annual rainfall levels are significantly lower than those required by other species such as palm oil, rape-seeds oil, sunflower oil, soybeans oil, corn oil and others. For instance, these plants can grow on all soil types in Indonesia, even on barren soil. The barren soil types can be found in many parts of eastern Indonesia that remain untapped because of the difficulty planted with other crops. Moreover, jatropha curcas oil is nonfood crops. Jatropha Curcas oil was processed by alkali base catalyzed esterification process using potassium hydroxide (KOH) to produce Jatropha Curcas methyl ester oil (JMEO) has a viscosity and a acidity that are acceptable for high voltage equipment especially in power transformer. The physicochemical and electrical properties of JMEO were measured as well as those of mineral oil (MO) for comparison. The physicochemical properties cover relative density, water content, viscosity, acidity, iodine number, corrosivity, flash point, pour point, color, visual examination, and methyl ester content. Meanwhile the electrical properties cover dielectric strength under AC, DC and lightning impulse voltages, pre-breakdown / streamers under lightning impulse voltage, creeping discharge over pressboard immersed in JMEO and MO. The obtained results show that the average DC and lightning impulse breakdown voltages of JMEO and MO are too close, even the average AC breakdown voltage of JMEO are higher than that of mineral oil (napthenic type). The measurement of breakdown voltages of two oil mixtures namely “80% JMEO + 20% MO” and “50% JMEO and 50% MO” shows that the breakdown voltage of the first mixture (i.e., “80%JMEO+20%MO”) is always higher than that of mineral oil under both AC and DC voltages. This indicates that mixing 20:80 mineral oil to JMEO ratio does not degrade its performance. The mixing of oils can occur when replacing mineral oil by JMEO in installed transformers. The analysis of the streamers characteristics (namely; shape, stopping length, associated current and electrical charge) developing in JMEO and MO under lightning impulse voltages, shows that these are too close (similar). It is also shown that the stopping (final) length Lf and the density of branches of creeping discharges propagating over pressboard immersed in Jatropha Curcas methyl ester oil (JMEO) and mineral oil (MO), under positive and negative lightning impulse voltages (1.2/50 μs), using two divergent electrode configurations (electrode point perpendicular and tangential to pressboard), are significantly influenced by the thickness of pressboard. For a given thickness, Lf increases with the voltage and decreases when the thickness increases. Lf is longer when the point is positive than with a negative point. For a given voltage and thickness of pressboard, the values of Lf in mineral oil and JMEO are very close. It appears from this work that JMEO could constitute a potential substitute for mineral oil for electrical insulation and especially in high voltage power transformers.

La spectrométrie de masse est une technique analytique permettant de mesurer le ratio masse sur charge d’un ion. Cette technique, très répandue en chimie analytique permet d’élucider la composition moléculaire de mélanges complexes à partir de systèmes homogénéisés. De ce fait, toute l’information sur la distribution spatiale des molécules est perdue. L’imagerie par spectrométrie de masse (IMS) a été inventée afin de résoudre ce problème et permettre d’élucider la distribution spatiale de molécules cibles sur des sections tissulaires minces provenant de tissus biologiques tels que de mammifères ou de plantes. L’un des grands avantages de l’IMS est sa complémentarité à l’histopathologie, technique permettant de révéler la structure ainsi que la localisation de certaines biomolécules à partir de sections tissulaires minces. Cependant, cette dernière se limite principalement aux protéines et aux molécules pouvant avoir une interaction spécifique avec un anticorps. L’IMS permet la détection d’une vaste gamme de biomolécules allant des petits métabolites aux polymères de haut poids moléculaire. Parmi les biomolécules détectables par IMS, les lipides attirent de plus en plus l’attention des analystes. En effet, ils occupent différentes fonctions clés au sein des systèmes biologiques, autant structurales que métaboliques, comme constituants des parois cellulaires, acteurs de la signalisation cellulaires ainsi que dans le stockage d’énergie. Leur intérêt est d’autant plus important qu’aucune technique histologique classique ne permet actuellement de détecter de façon spécifique les différentes classes de lipides. De façon générale, l’IMS de lipides est effectuée en utilisant la désorption-ionisation laser assistée par matrice (MALDI). Ce procédé permet de révéler l’emplacement de ii différentes classes de molécules en exploitant l’affinité que ces dernières ont pour une matrice particulière. Au-delà du choix de la matrice, d’autres paramètres tels que le mode de déposition de la matrice, le choix des solvants ainsi que le type de lavage utilisé vont également affecter le type de molécules détectés lors d’une analyse MALDI. Malgré les très bonnes performances du MALDI pour l’analyse de lipides, ce mode d’analyse se limite souvent aux lipides polaires facilement ionisables. Les lipides neutres comme le cholestérol (CHO) et les triacylglycérols (TAGs) sont impliqués à différents niveaux de fonctions biologiques fondamentales. Ainsi, nous avons développé trois stratégies permettant l’analyse de ces lipides neutres et de faibles abondances comme les gangliosides, directement à partir de sections tissulaires minces par MALDI ainsi que par désorption-ionisation laser classique (LDI). L’implication du cholestérol en tant que molécule structurale et précurseur de la synthèse de diverses hormones et vitamines, en fait une cible de choix pour l’analyse par IMS. Historiquement, l’analyse du cholestérol par MALDI permettait de le détecter sous sa forme déshydratée. De ce fait, il était impossible de le distinguer des autres métabolites tels que ses esters qui produisaient le même fragment. Afin de permettre l’IMS du cholestérol intact nous avons développé une nouvelle technique de préparation d’échantillons reposant sur le dépôt d’une couche nanométrique d’argent (16±2 nm) sur une section tissulaire mince par pulvérisation. Cette technique permet d’ioniser spécifiquement le cholestérol intact ainsi que divers acides gras sous forme d’adduits d’argent, et ce, avec une haute résolution spatiale (5 µm). iii Au-delà du cholestérol et des acides gras, une autre classe de lipides neutres très abondants, les triglycérides, reste difficilement analysable par MALDI IMS. En effet, les TAGs constituent la principale classe de lipides impliqués dans le stockage énergétique au niveau cellulaire. Ce rôle comme source d’énergie fait des TAGs un acteur incontournable de plusieurs maladies métaboliques telles que la stéatose hépatique, l’athérosclérose ainsi que la maladie d’Alzheimer. La difficulté d’analyser les TAGs par IMS provient de leur fragilité en milieu acide ainsi que de leur faible tendance à former des adduits sodium nécessaire à leurs analyses. En considérant ces limites, nous avons développé une méthodologie de préparation d’échantillon en deux étapes permettant l’analyse hautement spécifique des TAGs par LDI IMS. Dans un premier temps, les sections tissulaires minces sont initialement exposées à une solution aqueuse contenant un tampon carbonate à base de sodium (pH 10.3, 85 mM) ainsi que d’acétate de sodium (250 mM) afin de facilité la formation d’adduit sodium et de limiter la fragmentation des TAGs en source. Par la suite, une couche nanométrique d’or (28±3 nm) est déposée sur la section afin de permettre l’analyse des TAGs par IMS à haute résolution spatiale (> 10 µm). Lorsque ces derniers ont une abondance réduite dans les sections tissulaires, cette méthode permet aussi l’analyse d’esters de cholestérol (CE). La maladie de Hunter est une maladie génétique caractérisée par l’accumulation de glycosaminoglycanes (GAGs) ainsi que de l’accumulation secondaire de gangliosides. Ce phénomène est dû à l’absence de l’enzyme iduronate-2-sulfatase (IdS) qui permet la dégradation des GAGs. L’accumulation des GAGs et des gangliosides a pour conséquence l’apparition de problèmes fonctionnels et neurologiques majeurs entrainant la mort. Il existe une thérapie de remplacement enzymatique où une forme recombinante de l’enzyme IdS est injectée aux patients. Cette thérapie permet de rétablir le métabolisme normal des GAGs et iv gangliosides dans tous les organes sauf le cerveau où la barrière hémato-encéphalique empêche l’IdS recombinante d’atteindre les zones affectées. L’étude de la composition moléculaire des dépôts de GAGs et gangliosides au niveau cérébral constitue un défi important afin de comprendre la progression des troubles neurologiques engendrés par cette accumulation. À cette fin, nous avons développé une méthode MALDI spécifique à l’analyse des gangliosides à partir de sections tissulaires minces de cerveau de souris simulant la maladie Hunter (IdS-KO). Cette méthode d’analyse par MALDI IMS permet une révélation immuno-histochimique (IHC) des dépôts suivant l’analyse IMS. Nous avons pu visualiser cinq types de gangliosides dont quatre spécifiques au dépôt présent dans les cerveaux révélés par IHC sur la même section tissulaire. Cette étude nous a permis de distinguer pour la première fois des GM3 et GM2 selon la composition de leur chaine latérale et non de leur chaine polysaccharidique révélée par l’analyse IHC. / Mass spectrometry (MS) is an analytical technique that measures the mass-to-charge ratio of ions. This technique is widely used in analytical chemistry to solve the molecular composition of complex homogenized samples. The use of homogenized samples means that all the information with respect to the initial distribution of analytes is lost. Imaging mass spectrometry (IMS) is an MS technique which is able to provide the spatial localization of a given analyte on a surface, such as thin tissue sections from various animal sources. One of the greatest advantages of IMS is its complementarity with histopathology which normally reveals the general structure of thin tissue sections as well as the localization of certain biomolecules such as proteins or of any molecules capable of specific interactions with an antibody. On the other hand, IMS is capable of imaging a wide variety of biomolecules ranging from small metabolites to the high molecular weight proteins and polymers. Among these, lipids are of particular interest for their key involvements in many biological processes. Their interest is even greater when considering that lipid imaging by classical histology is unable to differentiate between all lipid species. IMS of lipids is typically performed using matrix assisted laser desorption/ionization (MALDI). MALDI IMS can differentiate various classes of lipids and their localization within a thin tissue section by taking advantage of the specific affinity that different classes of lipids have for different matrices. The matrix deposition process, along with the choice of solvents, are key parameters that need to be considered in MALDI IMS. While MALDI offers great coverage of the phospholipidome, it fails miserably for neutral lipid analysis. Indeed, cholesterol and triacylglycerols (TAGs) are two classes of neutral lipids with very important vi biological roles which are extremely difficult to image by MALDI. We have developed three new strategies that enables the detection of neutral lipids, some of which are expressed in low abundance such as gangliosides, directly from thin tissue section using either MALDI or laser desorption/ionization (LDI). Cholesterol is a precursor of many key biomolecules such as vitamins and hormones. It’s also a major component of the cellular membrane. MALDI IMS allows in some cases imaging of the dehydrated form of cholesterol. Unfortunately, detecting cholesterol as such makes it impossible to distinguish some of its metabolites which ionize in a similar fashion and dissociate to produce the same ions. To address this issue, we have developed a new sample preparation method involving the deposition of a nanometer scale silver layer (16±2 nm) over a thin tissue section. This enables the detection by LDI MS of intact cholesterol and some fatty acid species as silver adducts with up to 5 µm in spatial resolution. Beyond cholesterol and fatty acids, TAGs is another class of highly abundant neutral lipids still poorly detected by MALDI IMS. As TAGs are the main molecules involved in energy storage of cells, they have been implicated in many metabolic diseases such as non- alcoholic fatty liver disease, atherosclerosis and even Alzheimer’s disease. The reason why TAGs are poorly detected by MALDI comes from two key factors. First, TAGs are unstable in acidic environments, typical of MALDI matrices. Second, competition effects for the ionizing proton provided by the MALDI matrix prevent TAGs from easily ionizing through this main ionization process. To overcome these limitations, we have developed a new two-step sample preparation method for TAG LDI IMS. We initially deposited a solution of carbonate buffer (pH 10.3, 85 mM) and sodium acetate (250 mM) on the tissue section to increase the amount vii of available sodium for enhanced TAG ionization. The second step consisted of sputtering a nanometer scale UV absorbing gold layer (28±3 nm) that allows for the detection of TAGs by LDI IMS with spatial resolution as low as 10 µm. When TAGs are present in low amounts in the tissue section, this method also enables the detection of cholesterol esters. Hunter’s disease is a genetic disease characterized by the abnormal accumulation of glucoaminoglycans (GAGs) and the secondary accumulation of gangliosides due to the lack of iduronate-2-sulfatase (IdS) enzyme which controls their degradation. The accumulation of both GAGs and gangliosides form deposits which induces various functional issues to different organs as well as neurologic disorders. To minimize these effects, an enzyme replacement therapy has been developed. Unfortunately, it shows efficacy in all organs except the brain due to the inability of the recombinant enzyme to cross the blood-brain barrier. To further our knowledge of the progression of the disease, using a mouse model of Hunter’s disease we have developed a MALDI based method to specifically image gangliosides in brain deposits with a spatial resolution of 5 µm. This method also permits subsequent ganglioside staining by immunohistochemtry of the tissue section. With this method, we have identified four types of ganglioside which are specific to the Hunter’s disease pathology. We were also able to detect two types of deposits, one which is enriched in short chain gangliosides and the other in long chain gangliosides.

It has long been taken for granted that no serious drama work was done in Swedish schools before the 1950s. However, at the Tyringe Helpension – a progressive education girls’ boarding school that existed between the years 1909-1936 – drama was used as a method in many school subjects, as well as for social training. Ester Boman, the founder and principal of Tyringe, even talked about theatre as an experimental laboratory of the humanities. This study explores how that drama work evolved and why it has been forgotten. The study uses traditional history research methods with an emphasison hermeneutics, and with some addition from recent critical text analysis. The educational drama at the Tyringe Helpension is contextualized from five aspects: 1) The life and work of Ester Boman. 2) The private Swedish girls’ school system. 3) The international and the Swedish progressive education movement – Ester Boman was strongly influenced by Sofi Almquist and Ellen Key. She was also a member of the New Education Fellowship. 4) The teaching methods of the Tyringe Helpension as a whole. 5) Previous and contemporary use of drama in education and the theory of the dramatic instinct. The study shows that these five contexts were all important for the evolution of educational drama at the Tyringe Helpension and contributedin making the drama work there exceptionally rich and varied. These contexts are also crucial for the explanation of why this work was so quickly forgotten. However, it also had some importance that the Swedish drama pioneers of the 1950s were not particularly interested in what had been done before

The conception of electrospinning can trace its roots back more than 400 years, when it was observed that rubbed amber can deform a droplet of water on a smooth surface, and is based upon simple concepts of charge separation and surface tension. Since that time, considerable effort has been directed at both the cause and utility of this phenomenon. The specific aim of this dissertation project was to develop an automated electrostatic processing apparatus that was capable of controlling the three-dimensional architecture of an electrospun scaffold to further improve its utility in tissue engineering. The efficacy of using this technique has been well documented and can be adapted to produce tissue engineering scaffolds for a variety of tissues and organs. This apparatus incorporates precise mandrel motion. The system is capable of 0 - 5000 revolution per minute rotation, 0 - 25 inch per second translation and ± 40° rotation about the electrospinning jet axis for repeatable scaffold production. Fiber alignment and scaffold density are precisely controlled by rotating a mandrel along one axis, translation along that same axis, and rotation around the second axis perpendicular to the electrospun fiber stream. The control is accomplished with a PC based "supervisory" control program written partially in the LabVIEW® programming language and partially in SI Programmer supplied by Applied Motion Products. Scaffold thickness and fiber diameters are determined by the syringe metering pump flow rate, material being electrospun and solution concentrations. Through extensive laboratory analysis (mechanical testing and both optical and electron microscopy), parameters such as fiber orientation, diameter and mechanics can be predictive from specific polymer setups. Our laboratory has demonstrated the ability to electrospin natural and synthetic polymers and this apparatus will be utilized to tailor scaffolds to meet specific tissue engineering needs by creating a truly biomimicking scaffold / extracellular matrix.

Denna uppsats undersöker maktförhållandena i böckerna En piga bland pigor av Ester Blenda Nordström och Ett pennskaft som piga av Bonn i Taninge. Som metod och analysverktyg används intersektionellt perspektiv med utgångspunkt från de tre kategorierna kön, klass och funktionalitet. Vidare diskuteras maktförhållandena utifrån socialt- och genuskritiskt perspektiv för att spegla kvinnors och mäns villkor i det tidiga 1900-talets Sverige. Sammantaget visar litteraturanalysen att Ester Blenda Nordström besitter en maktposition även om hon utger sig för att vara en piga bland pigor. Den historiska kontexten som böckerna är skrivna i kännetecknas av modernitet och nationalskapande, och Ester Blenda Nordström blir rollmodell för den nya kvinnan som tar sig in på manliga domäner och kämpar för kvinnans rätt och jämlikhet i samhället.

A matéria-prima na produção de biodiesel corresponde a mais que 70% do seu custo e o estudo de viabilidade tecnológica e econômica das diferentes matérias-primas se reveste de enorme importância. A extração do óleo de soja com solvente etanol resulta em duas miscelas, uma rica em óleo (miscela rica) e outra rica em etanol (miscela pobre). A miscela pobre pode ser reutilizada no processo de extração e a miscela rica pode ser utilizada diretamente sem a necessidade de dessolventização e de etapas de refino. A miscela rica em óleo foi esterificada por dois processos diferentes: químico e enzimático, com diferentes concentrações em razão molar (óleo:etanol), diferentes temperaturas e catalisadores básico (NaOH) ou enzimático (Novozym®435), buscando o maior rendimento em ésteres etilícos. O objetivo desse estudo foi avaliar o rendimento de ésteres etílicos aplicando enzimas imobilizadas Novozym®435 e um catalisador básico (NaOH) e analisar a viabilidade energética da produção de biodiesel a partir da transesterificação da miscela rica (óleo:etanol) em óleo de soja, sem necessidade de refino do óleo. Foi adotado o planejamento experimental e análise de superfícies de resposta para a seleção das melhores condições de processo, tendo como variáveis respostas o rendimento e a qualidade do biodiesel. Foram realizados ensaios de esterificação via enzimática e química. A reutilização da enzima foi estudada através da lavagem com diferentes solventes (etanol 96%, isopropanol e terc-butanol) e reações de transesterificação na presença do co-solvente terc-butanol. A produção de ésteres em miscelas permitiu a comparação dos custos entre o processo de catálise enzimática e catálise química com base na análise dos fluxos de materiais e energia. A miscela rica foi obtida após três banhos com miscela pobre e um último banho com etanol 99% (v/v), apresentando eficiência de 83% e um teor de óleo residual no farelo de 4,2%. Em sua composição, a miscela rica apresentou 90% em óleo de soja e até 7% de etanol. A transesterificação de miscela rica com catalisador NaOH foi otimizada e apresentou rendimento de ésteres etílicos (RE) 97,2% nas condições experimentais de: razão molar 1:12, concentração de catalisador 0,67% e temperatura de 30ºC. Na transesterificação enzimática, o rendimento máximo foi de 85% nas condições reacionais de razão molar 1:4,5, concentração de catalisador 9,5% e temperatura de 40ºC. A Novozym®435 não foi recuperada com sucessivas lavagens dos solventes. Entretanto, o terc-butanol como co-solvente aumentou o rendimento de ésteres para 94%. A análise dos fluxos de energia demonstrou que o a obtenção da matéria-prima (laminação e extração) foi a etapa que mais demandou energia. A produção de miscela rica em escala semi-piloto demandou mais energia que a de óleo refinado, porém, a etapa de transesterificação a partir de miscela rica, utilizando o catalisador químico, demandou menos energia comparada ao processo com catalisador enzimático e o convencional com metanol e etanol. A esterificação de miscela rica é energeticamente viável, entretanto, com um scale up adequado, a etapa de extração com etanol deve ser ajustada para viabilizar energeticamente a cadeia de produção de biodiesel por esta via alternativa. / The feedstock for biodiesel production represents more than 70% of the cost and technological and economic feasibility studies of different oil sources are of enormous importance. The extraction of soybean oil with ethanol solvent results in two miscella, one rich in oil (rich miscellae) and another rich in ethanol (poor miscellae). The poor miscellae can be reused in the extraction process and the rich miscellae can be used directly without dessolventizing and refining stages. The oil rich miscellae was esterified by two different processes: chemical and enzymatic, with different concentrations in the molar ratio (oil: ethanol), different temperatures and either basic catalyst (NaOH) or an enzyme (Novozym®435), searching for the highest production of ethyl esters. The study goal was to prove the feasibility of producing biodiesel from the transesterification of rich miscellae (oilethanol) in soybean oil, without oil refining and evaluating the performance of ethyl esters by applying immobilized enzymes Novozym®435 and a basic catalyst (NaOH). We adopted the experimental design and the surface response methodology for the best selection of process conditions, with the response variables the yield and the quality of biodiesel. Chemical and enzymatic esterification trials were conducted. The reuse of enzyme was studied by washing with different solvents (96% ethanol, isopropanol and tert-butanol) and the transesterification reaction in the presence of the co-solvent tert-butanol. The production of esters by enzymes in the miscellae allowed a comparison of the costs between the enzymatic and chemical catalysis process based on the energy flow analysis. The rich miscellae was obtained after three baths employing the poor miscellae and a last fourth bath with ethanol 99% (v/v), presenting efficiency of 83% and a residual meal oil content of 4.2%. In its composition, the rich miscellae showed 90% of soybean oil and up to 7% ethanol. The transesterification of the rich miscellae with NaOH catalyst was optimized and had a ethyl esters yield (RE) of 97.2% under the experimental conditions of: 1:12 molar ratio, catalyst concentration 0.67% and temperature 30° C. For the enzymatic transesterification, the maximum yield was 85% for the reaction conditions: molar ratio 1:4.5, catalyst concentration 9.5% and temperature 40° C. Novozym®435 was not recovered with successive washes of the solvents. However, the tertbutanol as a co-solvent increased the yield of esters to 94%. The energy flow analysis showed that obtaining the raw material (flaking and extraction) was the most energy demanding. The rich miscellae from the semi-pilot plant demanded more energy than the refined oil, however, the transesterification of the rich miscellae using chemical catalyst, required less energy compared to the enzymatic catalysis and the conventional process methanol and ethanol. The esterification of rich miscellae is feasible energetically, however, the extraction step with ethanol should be adjusted to enable energetically the chain of biodiesel production.

Este trabalho teve como objetivo sintetizar o palmitato de isopropila, éster emoliente, empregando como biocatalisador lipases microbianas imobilizadas em partículas de poli(estireno-co-divinilbenzeno) (STY-DVB-M) obtidas por meio da técnica de polimerização em suspensão e magnetizadas por co-precipitação de íons de Fe +2 e Fe +3 em meio básico. Inicialmente, a influência da concentração do agente de suspensão e da agitação na distribuição granulométrica do polímero sintetizado foi avaliada por planejamento experimental 2 2, com triplicata no ponto central. As polimerizações que resultaram nas maiores quantidades de partículas com tamanhos apropriados para utilização como suporte para imobilização (entre 80 e 24 mesh), foram obtidas empregando as seguintes condições experimentais: 1% de agente de suspensão (PVA) e 400 rpm de agitação. O suporte obtido foi utilizado para imobilizar a lipase de Candida rugosa (LCR) e lipase de Penicillium camemberti (LG) via adsorção física e os biocatalisadores resultantes aplicados em reações de esterificação do ácido palmítico com isopropanol em meio heptano. Para cada biocatalisador foi adotado um planejamento experimental estrela rotacional 22, com triplicata no ponto central para avaliar a influência da concentração de biocatalisador (% m/v) e da razão molar (ácido:álcool) no rendimento de esterificação. Nas condições otimizadas em 12 h de reação foram obtidos 75,60 e 88,53% de rendimento de esterificação, respectivamente, para a LCR e LG imobilizada em STY-DVB-M. A quantidade de água formada durante o bioprocesso não foi considerada fator relevante para interferir no progresso da síntese. O biocatalisador obtido pela LG imobilizada em STY-DVB-M foi empregado em biorreator de tanque agitado (280 mL), na condição ótima, em um experimento com ampliação de escala, atingindo 85,68% de rendimento em 12 horas de reação. No entanto, foi observado cisalhamento do suporte em função da agitação mecânica, optando-se pela realização do bioprocesso em biorreator de leito fixo (dimensões: 11 mm de diâmetro interno x 16,6 mm de comprimento) com recirculação do meio reacional (1,5 mL.min-1). A operação do sistema nesta configuração foi impossibilitada pela evaporação e/ou percolação do solvente. Assim, foram realizadas reações em reator de leito fixo em meio isento de solvente nas seguintes razões molares: 1:3, 1:4 e 1:6 (ácido:álcool), cujos melhores resultados foram obtidos na razão molar de 1:4, apresentando 56% de rendimento, 215,88 g.L-1 do palmitato de isopropila e produtividade de 26,87 g.L-1.h-1, decorridas 8h de reação com vazão de reciclo de 1,5 mLmin-1. O emprego de peneira molecular no reservatório do substrato proporcionou um acréscimo de aproximadamente 7% na formação do éster. Essa mesma condição experimental (1:4 em meio isento de solvente) foi testada em reator de tanque agitado, atingindo produtividade de 33,40 g.L-1.h-1 em 8h de reação. No entanto, em função da desvantagem do cisalhamento do suporte constatada neste tipo de biorreator, concluiu-se que o biorreator de leito fixo foi a configuração mais adequada, para a condução do bioprocesso estudado, apresentando viabilidade do processo enzimático empregando lipase imobilizada em suporte híbrido magnetizado para a síntese do palmitato de isopropila. / The aim of this study was to synthesize isopropyl palmitate, an emollient ester, using as biocatalyst microbial lipase immobilized on magnetic copolymer. The support was prepared by suspension polymerization technique using styrene and divinylbenzene monomers in the presence of magnetite particles synthesized by co-precipitation of Fe+2 and Fe+3. The influence of the suspending agent concentration and the mechanical agitation on the granulometric distribution of the synthesized polymer was assessed by a 22 factorial design with triplicate at central point. The statistical analysis indicated that the optimal experimental conditions to obtain the largest amount of particles with suitable size (between 80 and 24 mesh) to be used as immobilizing support were attained at 1% suspending agent (PVA) at 400 rpm stirring. The resulting support was used to immobilize Candida rugosa lipase (LCR) and Penicillium camemberti lipase (LG) by physical adsorption and applied in the esterification reaction of palmitic acid with isopropyl alcohol in presence of solvent (heptane). For each biocatalyst a central composite 22 experimental design with triplicate at central point was performed to determine the influence of biocatalyst concentration (% m/v) and molar ratio (acid: alcohol) on the reaction yield. Under the optimal conditions higher performance was achieved by LG immobilized on STY-DVB-M (88.53%) than LCR immobilized on STYDVB-M (75.60%) with shaking at 12 hours reaction. For both reactions the amount of water formed as byproduct was not found to be a relevant factor that interferes in the synthesis progress. The most activity immobilized biocatalyst (LG STY-DVB-M) was used in agitated tank bioreactor (280 mL), under optimized conditions, in an experiment with scale-up, reaching 85.68% yield in 12 hours. However, it has been observed that the support was low mechanical shear and therefore an attempted was made to run the reaction in a packed bed bioreactor (dimensions: 11 mm internal diameter x 16,6 mm length) with recirculation of the reaction medium (1,5 mL.min-1). The system could not be operated under this configuration because of solvent evaporation and/or percolation. Thereby, the reactions were carried out in packed bed reactor using solvent-free medium under the following molar ratios: 1:3, 1:4 and 1:6 (acid: alcohol). The best performance (56% yield, 215.88 g L-1 of isopropyl palmitate and 26.87 gL-1.h-1 of productivity) was attained running the reactor with substrate at molar ratio of acid to alcohol of 1:4 for 8 hours with recycle rate of 1,5 mL.min-1. The use of a molecular sieve in the substrate reservoir resulted in an increase of approximately 7% of ester formation. By comparison, esterification run under the same experimental condition (1:4 in solvent-free medium) was carried out in a stirred tank reactor and slight higher productivity (33.40 g.L-1.h-1) was achieved. However, due the shear limitation of the support, it was concluded that the most suitable configuration for this bioprocess is the packed bed bioreactor, thereby showing that the enzymatic process is feasible using lipase immobilized on a magnetized hybrid support for isopropyl palmitate synthesis.

O uso de drogas por mulheres em idade reprodutiva é considerado um grave problema de saúde pública mundial. O mecônio é a primeira excreção do recém-nascido e tem sido utilizado como uma matriz alternativa em análises toxicológicas. A extração em fase sólida (SPE) é um método amplamente utilizado para a purificação e concentração dos analitos em amostras biológicas no campo da análise forense. A maioria dos produtos de SPE convencionais requerem volumes relativamente grandes de solventes levando a um custo acrescido por amostra e um aumento no tempo de processamento da amostra. As ponteiras de extração com fase sólida modificada (DPX) foram utilizadas como uma alternativa aos cartuchos SPE tradicionais. A técnica combina eficiência e rapidez no procedimento de extração, com redução significativa no consumo de solvente e na quantidade de amostra. O objetivo desse estudo foi desenvolver e validar um método para a determinação de nicotina, cotinina, cocaína, benzoilecgonina, cocaetileno e éster metil anidroecgonina em amostras de mecônio usando DPX e cromatografia em fase gasosa acoplada à espectrometria de massa (GC/MS). Os resultados da validação indicaram uma extração eficiente, exata e precisa, com recuperação entre 50-98%, exatidão entre 92-106%, precisão intra-ensaio 4-12% e precisão inter-ensaio 6 a 12%. As curvas de calibração foram lineares com valores de R2 superiores a 0,99; os limites de detecção (LOD) variaram entre 2,5-15 ng/g e os limites de quantificação (LOQ) entre 10-20 ng/g. O método DPX-GC/MS mostrou ser eficaz para análise traços de drogas presentes em amostras de mecônio. Após desenvolvimento e validação, o método foi aplicado em 50 amostras de mecônio coletadas no berçário da Maternidade do Complexo Aeroporto (MATER) na cidade de Ribeirão Preto - São Paulo, Brasil. / Drug abuse by pregnant women is considered a serious public health problem worldwide. Meconium is the first excretion in newborns and has been used as an alternative matrix to evaluate in utero drug exposure. Solid phase extraction (SPE) is widely employed to prepare and cleanup samples in the field of forensic analysis. Most SPE products require large volumes of solvent, which culminates in longer sample processing times; increased cost per sample and higher limits of detection. Disposable Pipette Extraction tips (DPX) have been used as an alternative to traditional SPE cartridges. They combine efficient and rapid extraction with reduced solvent consumption. The purpose of this study was to develop and validate a method to determine nicotine, cotinine, cocaine, benzoylecgonine, cocaethylene and methyl ester anhydroecgonine in meconium using DPX and GC/MS. Validation results indicated extraction efficiency, ranged between 50-98%, accuracy 92-106%, intra-assay precision 4-12% and inter-assay precision 6 to 12%. Linear calibration curves resulted in R2 values greater than 0.99; limits of detection ranged from 2.5 - 15 ng/g and the limit of quantitation from 10 - 20 ng/g. The DPX-GC/MS method provided to selectively analyze trace concentrations of drugs in meconium samples. Finally, the developed and validated method was applied to 50 meconium samples collected at the nursery of Maternidade do Complexo Aeroporto (MATER) in the city of Ribeirão Preto - São Paulo, Brazil.