Diss. (sammanfattning) Stockholm : Karol. inst., 2004. / Härtill 4 uppsatser.

The research summarized in this thesis focuses on directed evolution and enzyme mechanism studies of two aldolases: 2-deoxyribose-5-phosphate aldolase (DERA) and fructose-6-phosphate aldolase (FSA). Aldolases are nature’s own catalysts for one of the most fundamental reactions in organic chemistry: the formation of new carbon-carbon bonds. In biological systems, aldol formation and cleavage reactions play central roles in sugar metabolism. In organic synthesis, aldolases attract great attention as environmentally friendly alternative for the synthesis of polyhydroxylated compounds in stereocontrolled manner. However, naturally occurring aldolases can hardly be used directly in organic synthesis mainly due to their narrow substrate scopes, especially phosphate dependency on substrate level. Semi-rational directed evolution was used in order to investigate the possibility of expanding the substrate scope of both DERA and FSA and to understand more about the relationship between protein structure and catalytic properties. The first two projects focus on the directed evolution of DERA and studies of the enzyme mechanism. The directed evolution project aims to alter the acceptor substrate preference from phosphorylated aldehydes to aryl-substituted aldehydes. Effort has been made to develop screening methods and screen for variants with desired properties.  In the study of enzyme mechanism where enzyme steady state kinetic studies were combined with molecular dynamic simulations, we investigated the role of Ser238 and Ser239 in the phosphate binding site and the possible connection between enzyme dynamics and catalytic properties. The other two projects focus on the directed evolution of FSA and the development of a new screening assay facilitating screening for FSA variants with improved activity in catalyzing aldol reaction between phenylacetaldehyde and hydroxyacetone. The new assay is based on a coupled enzyme system using an engineered alcohol dehydrogenase, FucO DA1472, as reporting enzyme. The assay has been successfully used to identify a hit with 9-fold improvement in catalytic efficiency and to determine the steady state kinetic parameters of wild-type FSA as well as the mutants. The results from directed evolution illustrated the high degree malleability of FSA active site. This opens up possibilities to generate FSA variants which could utilize both aryl-substituted donor and acceptor substrates.

Cancer est un terme qui rassemble plusieurs ensembles hétérogène de maladies et il est caractérisé par la perte de contrôle physiologique et la transformation maligne des cellules saines. Il est essentiel de comprendre le cancer de la biologie cellulaire afin d'identifier de nouveaux biomarqueurs pour le diagnostic précoce et la conception de nouvelles stratégies thérapeutiques. Reprogrammation métabolique est une caractéristique émergente de cancer, ce qui signifie que les cellules cancéreuses passent leur métabolisme de base pour répondre aux exigences accrues de la croissance et la division cellulaire. Par conséquent, explorer reprogrammant métabolique que les cellules cancéreuses subissent est une stratégie clé pour identifier de nouvelles cibles pour le traitement du cancer. Dans cette thèse, de nouvelles possibilités pour le traitement du cancer ont été explorés en analysant la reprogrammation métabolique de la tumeur. À cet égard, nous avons étudié et proposé voie des pentoses phosphates (PPP) enzymes cibles thérapeutiques putatifs contre les cancers du sein et du côlon. En outre, nous avons exploré le métabolisme de la glutamine dans les cellules du cancer du sein et les adaptations du réseau métaboliques qu'ils subissent dans le but de contourner la privation de glutamine et la déficience mitochondriale générale. Ainsi, le ciblage PPP est l'intérêt des chercheurs d'utiliser à la fois oxydantes et non oxydantes phases de cette voie métabolique comme une cible de médicament thérapeutique. Pour tester cela, nous inhibés bœuf PPP enzymes 6PGD dans les cellules cancéreuses du sein et G6PD dans les cellules du côlon.Nous avons effectué la caractérisation de la reprogrammation métabolique induite par l'inhibition de l'enzyme de bœuf PPP par l'ARN interferase (ARNi) silençage médiation, afin d'explorer le potentiel de cette enzyme comme une cible de médicament thérapeutique dans deux lignées de cellules de cancer du sein. Nous avons demontré que l'inhibition 6PGD a entraîné une diminution taux de prolifération, arrêt du cycle cellulaire et induction de l'apoptose médiée par l'activation de p53, en diminuant les capacités de formation mammosphere et le métabolisme altéré de carbone central par modulation de Warburg phenomenan et en améliorant le métabolisme de la glutamine. D'autre part, nous avons montré l'effet de l'inhibition de la G6PD sur la prolifération des cellules du cancer du côlon et du PPP est régulée par la disponibilité de la glutamine dans les cellules cancéreuses du côlon.De plus, nous avons caractérisé les adaptations métaboliques que les cellules cancéreuses du sein subissent la privation de glutamine ou lorsque les mitochondries sont fait défection. Nous avons effectué une analyse des flux métaboliques utilisant métabolomique et Fluxomique et nous avons utilisé la biologie des systèmes afin d'estimer une vision globale des modifications de flux dans différentes conditions de culture. Nous avons observé une augmentation du cycle de pyruvate avec privation glutamine, ce qui indique que le ciblage des enzymes de cette voie telle que l'enzyme malique pourrait être une approche prometteuse combinée à l'inhibition de l'enzyme de glutaminase. D'autre part, nous avons observé que mimant une hypoxie par des cellules de cancer du sein de traitement redirigée oligomycine pour augmenter la carboxylation réductrice. Considérant que l'hypoxie est une condition commune dans l'environnement de la tumeur, le ciblage mécanisme de carboxylation réductrice pourrait être une nouvelle stratégie de lutte contre le cancer. Collectivement, les résultats présentés dans cette thèse démontre l'importance du métabolisme de la prolifération des cellules cancéreuses et la survie. Ce travail met également en évidence l'importance de la biologie des systèmes se rapproche de comprendre les mécanismes moléculaires sous-jacents des maladies multifactorielles complexes afin de souligner de nouvelles cibles thérapeutiques potentielles. / Moreover, we characterized the metabolic adaptations that breast cancer cells undergo in the deprivation of glutamine or when mitochondria are defected. We conducted metabolic flux analysis using metabolomics and fluxomics approaches and we employed Systems Biology approaches in order to estimate a global view of flux alterations in different culture conditions. We observed an increased pyruvate cycle with glutamine deprivation, thus indicating that targeting the enzymes of this pathway such as malic enzyme could be a promising approach combined with inhibition of glutaminase enzyme. On the other hand, we observed that mimicking hypoxia by oligomycin treatment redirected breast cancer cells to increase reductive carboxylation. Considering that hypoxia is a common condition in the tumor environment, targeting reductive carboxylation mechanism could be a novel strategy to fight against cancer. Collectively, all the results provided in this thesis demosntrate the importance of metabolism in cancer cell proliferation and survival. This work also highlights the importance of Systems Biology approaches to comprehend the molecular mechanisms underlying complex multifactorial diseases in order to point out new potential therapeutic targets.

L’élimination des produits agrochimiques indésirables des eaux de drainage agricole dans les bassins versants peut être réalisée par des zones humides ouvragées. Les oxy-hydroxydes de fer sont parmi les composés les plus réactifs chimiquement dans les sols tempérés et pourraient être un support idéal en mélange avec un support traditionnel pour la végétation dans ces systèmes. Le présent mémoire expose un travail de recherche visant à tester l’efficacité d’un procédé novateur d’élimination du glyphosate par un filtre minéral planté qui utilise les propriétés d’un support de grande surface spécifique composé d’oxy-hydroxyde ferrique et de végétaux adaptés aux zones humides et tolérants au glyphosate. Ce travail présente trois axes principaux qui sont : • la caractérisation physico-chimique de deux supports synthétiques choisis pour leur potentiel à retenir le glyphosate et de choisir le plus approprié pour notre étude • une étude mécaniste approfondie de l’adsorption du glyphosate sur le support retenu a été faite, suivie d’une analyse comparée de l’adsorption de glyphosate avec celle du phosphate sur ce même support. • la culture de différentes associations à partir de deux espèces végétales de milieu humide (Lythrum salicaria et Phragmites australis) sur un mélange de (sable, support adsorbant) arrosées par irrigation en présence ou en absence de glyphosate et de phosphate afin de vérifier l’efficacité du dispositif sur la dégradation de glyphosate. Les résultats obtenus de la caractérisation nous ont dirigés vers le minéral synthétique GEH®104, (GEH Wasserchemie GmbH & Co. KG, Heinrich-Hasemeier-Straße 33, D-49076 Osnabrück) qui est presque une akaganeite pure avec une surface spécifique de 350 m2.gr-1. Les cinétiques d’adsorption ont suggéré que le temps d’équilibre était plus long (15 j) que le temps déclaré dans la littérature pour des phases similaires. Cela est attribué à la nature mésoporeuse du support utilisé. L’akaganeite adsorbe le glyphosate fortement suivant deux modes : le premier est presque irréversible suivi par un mode hyperbolique, suivant l’équation de Langmuir double, alors que l’adsorption du phosphate suit plutôt l’équation de Bradley. La grande capacité d’adsorption de glyphosate et de phosphate par l’akaganeite en fait un candidat approprié pour une implication dans des zones humides ouvragées devant traiter des eaux contenant ces polluants. Les expériences de culture ont montré que Phragmites australis et la Lythrum Salicaria L. peuvent supporter de fortes concentrations de glyphosate par irrigation et qu’ils bénéficient de l’association mutuelle. Il semblerait que les plantes et leurs micro-organismes associés assurent l’essentiel de la disparition du glyphosate en solution car une fois bien installées, le glyphosate et ses métabolites ne sont pratiquement plus détectables en solution. Le système complet empêche le glyphosate de percoler lorsque celui-ci est présent avec ou sans phosphate dans les eaux d’irrigation / The elimination of undesirable agrochemicals of agricultural drainage water in watersheds can be achieved through engineered wetlands. Iron oxy-hydroxides are among the most chemically reactive compounds in temperate soil and they could be an ideal support in admixture with a traditional holder for vegetation in these systems. This work aimed to test the effectiveness of an innovative method of removing glyphosate mineral planted by a filter that uses the properties of a high surface area carrier composed of ferric oxyhydroxide and plants adapted to wetlands and tolerant to glyphosate. This work has three main axis which are: • The physicochemical characterization of two synthetic materials chosen for their potential to retain glyphosate and choose the most suitable for our study. • A profound mechanism study of the adsorption of glyphosate on the chosen support has been made, followed by a comparative analysis of the glyphosate with the adsorption of phosphate on the same support. • culture of different associations based on two plant species of wetland (Lythrum salicaria and Phragmites australis) on a mixture of (sand, adsorbent support) watered by irrigation in the presence or absence of glyphosate and phosphate to verify the effectiveness of the device on the degradation of glyphosate. The results of the characterization directed us to the synthetic mineral GEH®104 (GEH Wasserchemie GmbH & Co. KG, Heinrich-Hasemeier Straße 33, D-49076 Osnabrück) which is almost a pure akaganeite with a surface area of 350 m2.gr-1. The adsorption kinetics suggested that the equilibrium time was longer than (15 days), which is longer than the time reported in the literature for similar phases. This is attributed to the mesoporous nature of the support used. Akaganeite strongly adsorbs glyphosate in two modes: the first is almost irreversible followed by a hyperbolic mode, according to the Dual Langmuir’s equation while the phosphate adsorption follows the Bradley’s equation. The large capacity of adsorption of glyphosate and phosphate by actually akaganeite makes it a suitable candidate for involvement in wetlands engineered to treat water containing these pollutants. The experimental cultural showed that reed and purple loosestrife can support high concentrations of glyphosate by irrigation and they benefit from membership organization. It appears that plants and their associated microorganisms are responsible for most of the disappearance of glyphosate in solution because once properly installed, glyphosate and its metabolites are practically no longer detectable in solution. The complete system prevents glyphosate from percolating when it is present with or without phosphate in the irrigation water

128f. / Submitted by Ana Hilda Fonseca (anahilda@ufba.br) on 2013-04-01T15:08:24Z No. of bitstreams: 1 Tese Raildo.pdf: 62587482 bytes, checksum: 8ffed170e1190fc01d8858d45dc24efd (MD5) / Approved for entry into archive by Ana Hilda Fonseca(anahilda@ufba.br) on 2013-04-24T15:41:17Z (GMT) No. of bitstreams: 1 Tese Raildo.pdf: 62587482 bytes, checksum: 8ffed170e1190fc01d8858d45dc24efd (MD5) / Made available in DSpace on 2013-04-24T15:41:18Z (GMT). No. of bitstreams: 1 Tese Raildo.pdf: 62587482 bytes, checksum: 8ffed170e1190fc01d8858d45dc24efd (MD5) Previous issue date: 2010 / CAPES / Nesse trabalho foram desenvolvidos métodos analíticos para a determinação de metais e especiação de arsênio em amostras de fertilizantes fosfatados e fosfato de rocha. Para isso, foi realizado um estudo de diâmetros e distribuição de partículas nas amostras e verificou-se que o preparo da suspensão promoveu diminuição do diâmetro médio das partículas. Foi desenvolvido um método para a determinação de Cd, Cr, Cu, Mn, Co, Ni, Fe e Zn empregando amostragem de suspensão e determinação por espectrometria de emissão ótica com plasma indutivamente acoplado (ICP OES). As condições de preparo da suspensão foram otimizadas utilizando metodologia univariada e as melhores condições foram: 200 mg de amostra, 20 mL de HCl 0,5 mol L-1 e tempo de sonicação de 25 minutos. A precisão do método foi avaliada pelo desvio padrão relativo, RSD (%) e os valores foram sempre inferiores a 9,2 %. No teste de adição e recuperação na amostra SSP1, os valores de recuperação para os analitos foram sempre superiores a 90% e inferiores a 109%. A exatidão do método foi avaliada pela comparação do método de amostragem de suspensão com o método oficial de determinação de metais e metalóides em fertilizantes da Association Official Analytical Chemists (AOAC) e aplicando-se análise de variância (ANOVA) fator único, não foram observadas diferenças significativas entre as concentrações dos analitos determinadas pelos dois métodos. Foi aplicada análise multivariada como proposta metodológica para comparação de métodos analíticos e com base na avaliação da análise de componentes principais (PCA) e da análise de agrupamentos hierárquicos (HCA), verificou-se que a análise multivariada é uma ferramenta que pode ser utilizada com sucesso para comparação de diferentes métodos analíticos. Os resultados da ANOVA fator único correlacionaram muito bem com os resultados obtidos por PCA e HCA. Outro método foi desenvolvido para a determinação de As em fertilizantes fosfatados e fosfato de rocha também empregando amostragem de suspensão e geração de hidretos acoplada à espectrometria de absorção atômica (HG-AAS). As amostras foram submetidas à extração em meio ácido clorídrico 6 mol L-1 e submetidas a banho de ultra-som por 30 minutos. A determinação de As total foi obtida mediante etapa de redução da espécie As (V) a As (III) utilizando KI e ácido ascórbico e a determinação de As (III) pela adição de ácido cítrico/citrato de sódio, que mascara a espécie As (V). Desta forma, a concentração de As(V) é encontrada pela diferença entre as concentrações de As total e As (III) na amostra. Nas condições experimentais estabelecidas, este método permite a determinação de As total com limite de detecção e de quantificação iguais a 0,01 e 0,03 mg kg-1, respectivamente. A precisão expressa RSD(%), foi sempre menor que 8%. A exatidão do método foi avaliada pela determinação de As total na amostra após digestão ácida em bloco de aquecimento e sistema de refluxo a determinação em dois materiais de referência certificado de sedimento marinho, MESS-2 e MESS-3. / Salvador

Made available in DSpace on 2014-06-11T19:31:36Z (GMT). No. of bitstreams: 0 Previous issue date: 2004-04-22Bitstream added on 2014-06-13T21:02:54Z : No. of bitstreams: 1 mendonca_cctn_dr_botfca.pdf: 609115 bytes, checksum: 61293a5052971a6e02dc4478d68dbc4f (MD5) / Universidade Estadual Paulista (UNESP) / O processo de solubilização, dissolução de minerais do solo e suas interações com os processos de adsorção e dessorção de fosfato são de alta importância devido às possibilidades de otimização no processo de adubação do solo para o plantio. Alguns autores sugerem que a dissolução de ferro nos solos ocorre por redução microbiana e complexação, interferindo diretamente no processo de adsorção e dessorção do fosfato. O objetivo deste trabalho foi investigar a relação existente entre liberação de ferro, a neoformação de óxidos de ferro e adsorção e dessorção de fosfato em um Latossolo Vermelho distrófico de região Tropical Úmida. A relação foi estudada em solos sob diferentes tipos de ocupação: cana de açúcar, café, milho, reflorestamento de eucalipto e floresta natural. Amostras de solo foram coletadas nos horizontes A e B, tratadas com solução de sacarose a 3% e mantidas em repouso. O processo redox ocorreu em todas as amostras tratadas o que foi comprovado pela análise de pH onde se obteve uma acidificação do meio e posterior aumento do pH, indicando o estabelecimento de um equilíbrio químico do processo, mantendo assim o pH estável. Análises quantitativas de ferro, zinco, cobre, manganês e fósforo foram realizadas no solo natural, no solo sob tratamento e na fase líquida das amostras. A estes solos foram adicionadas duas concentrações de fosfato: 200 mg.dm3 e 400 mg.dm3 e calculadas as adsorções. A comparação dos resultados destas análises mostrou que a fixação de fósforo foi maior nos solos submetidos ao tratamento. Isto pode ser explicado pela decomposição microbiana dos compostos orgânicos, na interface com a atmosfera oxidante, que provocou a precipitação dos óxidos e hidróxidos dos metais onde os íons fosfato foram fixados pelos óxidos e hidróxidos reprecipitados. / The solubilization process, minerals dissolution and its interactions with the phosphate adsorption/desorption processes are of high importance due to the possibilities of improvement in the process of the soil manuring in the plant crops.Some authors suggest that the iron dissolution in the soils occurs by microbial reduction and complexation, interfering directly in the phosphate adsorption /desorption process.The purpose of this work was to investigate the existent connection between the iron liberation, the neoformation of iron oxides and the phosphate adsorption/ desorption in a Latossolo Vermelho Distróficoof humid tropical region.This connection was studied in soils by different crops: sugar cane, coffee, corn, reforestation with eucalyptus and natural forest. Soil's samples were collected in A and B horizon, treated with sucrose 3% solution and maintained in repose.Redox process occurred in all the treated samples, what was proved by the pH analysis, in where were obtained some acidification of the environment and subsequent pH increase, indicating the establishment of a chemical balance of the process, maintaining like this the pH stable. Quantitative analyses of iron, zinc, copper, manganese and match were accomplished in the natural soil, in the soil under treatment and in the liquid phase of the samples.Two phosphate concentrations: 200 mg.dm3 and 400 mg.dm3 were added to these soils and the adsorptions were calculated. The comparison of these analyses results shows that the phosphorous fixation was higher in the soils submitted to the treatment. This can be explained by the microbial decomposition of the organic compounds, in the interface with the oxidant atmosphere, which nettled the precipitation of the oxides and hydroxides of the metals in where the phosphate ions were fixated by oxides and hydroxides reprecipitated.

Made available in DSpace on 2014-07-29T15:12:49Z (GMT). No. of bitstreams: 1 dissertacao fernando damasceno.pdf: 2320631 bytes, checksum: 27fcd42bbb835c6752f9bc475e38b816 (MD5) Previous issue date: 2009-05-26 / Aluminum hydroxides are efficient phosphate adsorbents. In this work it was investigated the adsorptions of phosphate in three different forms of aluminum hydroxide (amorphous, bayerite and pseudo-boehmite). The adsorption isotherms present the amount of phosphate adsorbed as a functions of the equilibrium concentration of orthophosphate ions aqueous solutions, in pH = 5. The products of adsorption were characterized by infrared spectroscopy, 27Al e 31P NMR spectroscopy, XRD analysis, and by zeta potential measurements of the aqueous dispersions. The adsorption isotherms of phosphate on aluminum hydroxide showed that when the hydroxides are treated with diluted phosphate solutions (< 2,0 mmolL-1) the three hydroxides adsorb similar amount of phosphate. When the hydroxides are treated with concentrated phosphate solutions (> 2,0 mmolL-1) bayerite and amorphous aluminum hydroxide incorporate much more phosphate than pseudo-boehmite. The IR and the NMR 27Al and 31P analysis of the adsorption products showed that when the adsorption is carried out in low concentrate phosphate solutions, the adsorbed species are surface aluminum phosphate complexes, while in samples treated with concentrated phosphate solution the products of adsorption are solid phases of aluminum phosphate. These results suggest that the incorporation of phosphate by the three aluminum hydroxides studied occur in two steps. In the first step the phosphate are adsorbed yielding surface complexes; in the second step the incorporation of phosphate occur via the process of aluminum dissolution and precipitation of an aluminum phosphate phase. The XRD analysis showed that the precipitate formed during the dissolution-precipitation process is amorphous and become crystalline after heated at 750 °C for 30 min. The slope of the zeta potential measurements of the aqueous suspensions of the adsorption products suggests that the concentration of the phosphate solution at which the transition of phosphate incorporation by adsorption changes to phosphate incorporation by dissolution-precipitation is similar for the three hydroxides (2 mmolL-1). The results showed that the incorporation of phosphate by precipitation-dissolution occur on the three hydroxides but with lower extension on pseudo-bohemite. This behavior might by related to the lower dissolution of pseudo-bohemite because it is more condensed than amorphous aluminum hydroxide and bayerite / Hidróxidos de alumínio são conhecidos como bons adsorventes de fosfato. Nesse trabalho investigou-se a eficiência na incorporação de fosfato de três diferentes tipos de hidróxido de alumínio (amorfo, bayerita e pseudo-boehmita). O estudo foi realizado obtendo-se as isotermas de adsorção de fosfato nos três hidróxidos. Determinou-se o potencial zeta dos produtos das adsorções e caracterizou-se os produtos por espectroscopia na região do infravermelho, espectroscopia de RMN (31P e 27Al) e difratometria de raios X. As curvas de adsorção mostram que em concentrações baixas de fosfato (< 2,0 mmolL-1) os três hidróxidos de alumínio incorporam quantidades semelhantes de fosfato, enquanto que em concentrações altas de fosfato (> 2,0 mmolL-1) o hidróxido de alumínio amorfo e a bayerita incorporam quantidades de fosfato superior à quantidade incorporada pela pseudo-boehmita. A caracterização dos produtos da adsorção por IV e RMN revelou que quando se emprega soluções diluídas de fosfato, os produtos de adsorção são complexos superficiais de ortofosfato com alumínio, o que é indicativo de que a incorporação de fosfato se dá por adsorção. Nas amostras tratadas com soluções de fosfato concentradas foi observada a presença de uma fase de fosfato de alumínio, o que sugere que nestas condições, a incorporação de fosfato ocorre principalmente pelo processo de dissolução do hidróxido de alumínio seguida de precipitação de uma fase de fosfato de alumínio. As análises por DRX dos produtos da adsorção em altas concentrações de fosfato revelaram que a fase formada durante o processo de dissolução-precipitação é amorfa e transforma-se em AlPO4 cristalino quando submetida a aquecimento a temperatura de 750 °C por 30 min. As medidas de potencial zeta dos produtos de adsorção indicam que nos três hidróxidos estudados o processo de incorporação de fosfato se dá pelos mesmos processos: inicialmente, por adsorção de fosfato e em seguida por dissolução do hidróxido e precipitação de fosfato de alumínio. Os resultados mostram que a incorporação de fosfato por dissolução-precipitação ocorre nos três hidróxidos, mas com maior extensão no hidróxido de alumínio amorfo e na bayerita e com menor extensão na pesudo-boehmita. Esse comportamento pode estar relacionado com o fato da pseudo-boehmita ser o hidróxido de alumínio mais condensado e consequentemente menos susceptível ao processo de dissolução

Foram avaliados os efeitos do fósforo proveniente das fontes Fosfato Bicálcico (FBC) e Fosfato Monoamônico (FMA) sobre a digestibilidade ruminal in situ do feno de coast-cross (Cynodon dactylum) e a solubilidade das mesmas fontes de P, através de estudo in vitro. Doze carneiros machos, castrados, mestiços da raça Sulfock, dotados de cânulas ruminais, foram empregados por um período experimental dividido em duas etapas: a primeira avaliou a solubilidade in situ do fósforo proveniente das duas fontes em três diluentes distintos, sendo líquido ruminal obtido de animais recebendo feno e concentrados, líquido ruminal obtido de animais recebendo apenas feno e solução de ácido cítrico a 2%. Para quantificação da solubilidade do FMA e FBC no líquido ruminal foi formado um pool distinto dos líquidos de cada grupo quanto aos alimentos oferecidos. Em seguida as mostras foram dissolvidas nos diluentes e determinou-se a concentração de fósforo total no meio, a partir da leitura da absorbância em espectrofotômetro. A segunda etapa foi desenvolvida para avaliar o efeito da disponibilidade do fósforo das duas fontes sobre a digestibilidade ruminal in situ do mesmo feno de coast-cross. O delineamento experimental utilizado foi o inteiramente casualizado composto por dois tratamentos: Fosfato Bicálcico e Fosfato Monoamônico. A digestibilidade da matéria seca e da fibra em detergente neutro foi calculada pelo emprego da técnica dos sacos de náilon incubados no rúmen durante 6, 12, 24, 48, 72 e 96 horas. O fosfato monoamônico apresentou maior disponibilidade no líquido ruminal em relação ao bicálcico (P<0,05). No entanto, essa diferença na concentração de P disponível no líquido ruminal não promoveu aumento significativo na digestibilidade in situ da FDN ou da MS do feno. / The effects of phosphorus from Dicalcium Phosphate (DCP) or Monoammonium Phosphate (MAP) sources were evaluated on in situ ruminal digestibility of coast-cross hay (Cynodon dactylum) and also their solubility using in vitro study. Twelve Suffolk crossbred wethers were used, fitted with ruminal cannulas. The experimental period was divided into two stages: the first one evaluated the in situ solubility of the two sources of phosphorus in three different extractors: ruminal fluid from animals receiving roughage plus concentrate-based diet; ruminal fluid from animals receiving only roughage-based diet; and 2% citric acid solution. To quantify the solubility of DCP and MAP in rumen fluid, a pool of fluid from each group, relating to the feed offered, was formed. The samples were dissolved in solvents and total phosphorus concentration was determined, by reading the absorbance in a spectrophotometer. The second stage evaluated the effect of phosphorus availability from the two sources used on in situ ruminal digestibility of the coast-cross hay. A completely randomized design was used, composed by two treatments: Dicalcium Phosphate and Monoammonium Phosphate. Digestibility of dry matter and neutral detergent fiber was calculated, applying in situ nylon bag technique, with incubation time of 0, 6, 12, 24, 48, 72 or 96 hours. Monoammonium phosphate showed higher availability in ruminal fluid than dicalcium phosphate (P<0,05); however, this difference in the concentration of phosphorus available in ruminal fluid did not cause significant increase on in situ digestibility of NDF or DM of hay.

L'un des défis des recherches actuelles réside dans l'élaboration d'outils pharmacologiques capables de juguler la croissance tumorale en ciblant spécifiquement son angiogenèse. L'angiogenèse est un processus physiologique complexe qui fait intervenir de nombreux récepteurs, parmi lesquels se trouve le récepteur du mannose 6-phosphate/insulin like growth factor II (M6-P/IGFII). Des travaux antérieurs effectués au laboratoire ont montré que des molécules analogues du M6P sont des effecteurs d'angiogenèse mais qu'au cours de leur administration, ces composés sont facilement éliminés car trop hydrophiles. Afin d'obtenir des analogues du M6P plus lipophiles nous avons remplacé le méthyle en position anomère par d'autres chaines plus longues. Nous présentons, au cours de cette thèse, la synthèse d'analogues du mannose 6-phosphate et l'évaluation de leur activité angiogénique par la méthode « CAM». / One of the challenges of the current research is to develop pharmacological tools able to curb tumor growth by specifically targeting the angiogenesis. Angiogenesis is a complex physiological process that involves a large number of receptors, among which the mannose 6-phosphate/insulin-like growth factor II (M6-P/IGF-II) receptor. Previous work conducted in the laboratory has shown that analogues of M6P are effectors of angiogenesis but during administration, these compounds are easily removed due to their hydrophilicity. To increase the lipophilicity of the M6P analogues, we replaced the methyl in position anomeric by other longer chains.We present in this manuscript, the synthesis of mannose 6-phosphate analogues and the evaluation of their angiogenesis activity by the “CAM assay”.

Detta examensarbete har utförts på Ovako i Hällefors i syfte att undersöka möjligheterna till att fosfatera stålkutsar vid Ovako Sweden AB. Uppdraget har förmedlats av Ovako Sweden AB i Hällefors i syfte att ta fram ett koncept som kan användas för att fosfatera stålkutsar. Ovakos lösning på detta problem, i nuläget, är att skicka kutsarna till en firma i Tyskland där fosfateringen sker. Det som Ovako ser som nackdel med denna lösning är att när fosfateringsprocessen sker hos en extern leverantör uppkommer svårigheter med att säkerställa kvalitén på produkten. Detta innebär i sin tur att om Ovakos skulle få en reklamation från en kund, blir det svårt att avgöra om ytdefekten uppstod under fosfateringen hos leverantören i Tyskland, hos kunden eller hos Ovako. Målet med projektet är att genom teoretiska studier och eventuella praktiska försök ta fram ett lämpligt förslag på hur befintlig fosfateringslinje behöver byggas om eller modifieras för att möjliggöra fosfatering av stålkutsar enligt kundens önskemål. Det har formulerats fyra olika koncept som bygger på intervjuer, observationer och nulägesanalys vid sektionerna Drapro och Preco. För att få fram det lämpligaste konceptet har Pugh matris används som hjälpmedel. Det har även gjorts en kostnadskalkyl för koncept förslaget för det försatta arbetet. Det togs fram fyra olika koncept A, B, C och D. A och B gick inte författaren vidare med och med hjälp av Pugh-matrisen gjordes en jämförelse mellan koncept C och D. Slutresultatet av denna rapport är att koncept C har valts som den lämpligaste lösningen, för att den minskar risken för ytdefekter på kutsen samt att den uppfyller företagets krav. / This thesis has been carried out at Ovako in Hällefors in order to review the possibilities for phosphating steel pellets at Ovako AB, Sweden. The assignment has mediated by Ovako AB Sweden in Hällefors in order to develop a concept that can be used for phosphating steel pellets. Ovako's solution to this problem, is to send the pellets to a firm in Germany where phosphates take place. What Ovako sees as a disadvantage with this solution is that when the phosphate process handled by an external supplier, difficulties can arise in ensuring the quality of the product. This in turn means that if Ovako would receive a complaint from the customer, in which that it becomes difficult to determine if the surface defect occurred during phosphating by the supplier in Germany, if it’s the customer who is responsible for the surface defect or maybe it occur at Ovako during the production of the pellets. The goal of the project is that through theoretical studies and practical attempts to develop an appropriate proposal on how existing phosphating line needs to be modified to allow phosphating of steel pellets according to customer requirements. In order to find a proper solution has four different concepts formulated based on interviews, observations and current situation analysis on both sections Drapro and Preco. Pugh matrix has been used to obtain the most appropriate concept in this thesis. It has also made a cost estimate for the concept proposal for the added work. Four concepts were developed; A, B, C and D. The author didn’t proceed with concept A, B, and by using the Pugh matrix, a comparison was made between C and D. The final outcome of this report is that concept C has been selected as the most suitable solution, because it reduces the risk of surface defects on the pellet, and that it fulfills the company's requirements.