Orientador: Jaime Aparecido Cury / Tese (doutorado) - Universidade Estadual de Campinas, Faculdade de Odontologia de Piracicaba / Made available in DSpace on 2018-08-24T16:20:44Z (GMT). No. of bitstreams: 1 Botelho_JulianaNunes_D.pdf: 1920977 bytes, checksum: 99c53e9c14bfc15c6b8859cce1fe5dec (MD5) Previous issue date: 2014 / Resumo: Sacarose é o carboidrato mais cariogênico da dieta e o amido é considerado não cariogênico para esmalte e moderadamente cariogênico para dentina. Por outro lado, a combinação de amido e sacarose (amido+sacarose) tem sido considerada mais cariogênica que apenas sacarose, mas esse ainda é um assunto em debate. Além do mais, o efeito do dentifrício fluoretado na cariogenicidade dessa combinação é desconhecido. Assim, com o objetivo de estudar esse assunto três experimentos foram conduzidos: (i) o primeiro avaliou efeito de amido+sacarose na desmineralização de esmalte e dentina, usando um modelo de biofilme de S. mutans modificado pela adição de saliva para simular a ação da amilase, (ii) o segundo avaliou in situ o efeito do dentifrício contendo 1.100 µg F/g (DF) na progressão da desmineralização da dentina radicular, e o terceiro (iii) avaliou in situ o efeito do fluoreto no potencial cariogênico de amido+sacarose na desmineralização de esmalte e dentina. In vitro, biofilmes de S. mutans foram formados sobre blocos de esmalte e dentina radicular, por 5 e 4 dias respectivamente, em meio de cultura contendo saliva e expostos a um dos seguintes tratamentos: amido a 1%, sacarose a 10% ou de sua combinação (8x/dia). Os biofilmes foram analisados quanto às suas composições bioquímicas e microbiológicas, e a desmineralização dos blocos foi avaliada. Biofilmes expostos à combinação foram mais acidogênicos (p<0,0001) e provocaram maior desmineralização (p<0,0001) no esmalte e dentina que o efeito dos carboidratos isolados. In situ, o efeito do DF foi testado em um estudo piloto, cruzado no qual sacarose a 10% foi aplicada extraoralmente 8x/dia em 2 fases de 14 dias. Após 10 e 14 dias em cada fase, a desmineralização da dentina foi avaliada. O efeito do dentifrício foi significativo (p<0,0001), mas o efeito do tempo não (p>0,05). Esses resultados sugerem que o DF com 1.100 µg F/g é capaz de diminuir a cárie dentinária mesmo sob alto desafio cariogênico de acúmulo de biofilme e exposição à sacarose. In situ, o efeito dos tratamentos (água, amido a 2%, sacarose a 10% e amido+sacarose) e o efeito do dentifrício (não fluoretado e fluoretado) foram testados em um estudo cruzado, cego, boca-dividida em 4 fases de 14 dias. Os voluntários usaram dois dos tratamentos 8x/dia e um dos dentifrícios 3x/dia. O efeito dos fatores (dentifrício e tratamentos) foram significativos (p<0,05) para esmalte e dentina, mas a interação não (p>0,05). Os resultados sugerem que, independente do desafio cariogênico provocado pelos diferentes carboidratos da dieta testados, o dentifrício fluoretado é efetivo na redução da desmineralização de esmalte e dentina. Em conclusão, os resultados sugerem que amido deve aumentar o potencial cariogênico da sacarose mas que fluoreto de dentifrício é capaz de reduzir a desmineralização tanto do esmalte quanto da dentina provocada pela combinação desses carboidratos / Abstract: Sucrose is the most cariogenic dietary carbohydrate while starch is considered non-cariogenic for enamel and slightly cariogenic for dentine. The combination starch and sucrose (starch+sucrose) has been considered more cariogenic than sucrose alone but this subject remains debatable. Also, the effect of fluoride toothpaste on the cariogenicity of this combination is unknown. The aims of this study were to evaluate: (i) the effect of starch+sucrose on enamel and dentine demineralization using an S. mutans biofilm model modified by adding human saliva to simulate amylase action; (ii) the in situ effect of fluoride toothpaste (FT) containing 1100 µg F/g on dentine demineralization progression; and (iii) the in situ effect of fluoride on the cariogenic potential of starch+sucrose on enamel and dentine demineralization. In vitro, S. mutans biofilms were grown on enamel and root dentine slabs for 5 and 4 days, respectively, in a saliva-containing medium and exposed to the following treatment: 1% starch; 10% sucrose; or starch+sucrose (8x/day). Biofilms were then analyzed for their biochemical and microbiological compositions, and dental demineralization was evaluated. Biofilms exposed to starch+sucrose were more acidogenic (p < 0.0001) and caused higher demineralization (p < 0.0001) on either enamel or dentine than those exposed to each carbohydrate alone. The in situ effect of FT on dentine demineralization was tested in a pilot crossover study, in which 10% sucrose was applied extra-orally to the slabs 8x/day in 2 phases of 14 days each. At days 10 and 14 of each phase, dentine demineralization was evaluated. The effect of toothpaste was significant (p<0.0001), but the effect of time was not (p>0.05). The results suggest that FT at 1100 µg F/g can reduce dentine demineralization even under high cariogenic challenges - biofilm accumulation and sugar exposure. The in situ effect of the treatments (water, 2% starch, 10% sucrose and starch+sucrose) and that of the toothpastes (non-FT and FT) were tested in a crossover, single-blind and split-mouth study conducted in 4 phases of 14 days each. The volunteers used two of the treatments 8 times/day and one of the toothpastes 3 times/day. The effect of the factors (toothpaste and treatments) was significant (p<0.05) for enamel and dentine, but not (p>0.05) for the interaction. The findings suggest that, regardless of the cariogenic challenge provoked by the different sources of the dietary sugars tested, fluoride toothpaste is effective in reducing enamel and dentine demineralization. In conclusion, the results suggest that starch may enhance the cariogenic potential of sucrose and fluoride from toothpaste reduces enamel and dentine demineralization caused by the combination of these carbohydrates / Doutorado / Cariologia / Doutora em Odontologia

Orientador: Marcos Pereira Marinho Aidar / Dissertação (mestrado) - Universidade Estadual de Campinas, Instituto de Biologia / Made available in DSpace on 2018-08-26T03:32:16Z (GMT). No. of bitstreams: 1 Pikart_FilipeChristian_M.pdf: 1633217 bytes, checksum: 40896cc3191e7fb0cbac5cb3699d7b31 (MD5) Previous issue date: 2014 / Resumo: As espécies epífitas estão submetidas a períodos de escassez hídrica devido ao regime intermitente de chuvas e à falta de acesso à água do solo. Entretanto, algumas espécies apresentam estratégias para a manutenção do seu conteúdo hídrico, tais como a expressão do Metabolismo Ácido das Crassuláceas (CAM). Guzmania monostachia é uma bromélia epífita que apresenta regulação positiva do CAM quando submetida a um período de deficiência hídrica. Em trabalho anterior, após sete dias de restrição hídrica, a espécie apresentou um CAM do tipo idling, caracterizado pelo fechamento estomático durante os períodos claro e escuro associado ao acúmulo noturno de ácidos orgânicos. Também foi observado um gradiente funcional ao longo das folhas, onde o ápice apresentou maior número de estômatos, de pigmentos relacionados à fotossíntese e da atividade das enzimas PEPC e MDH, sugerindo maior atividade fotossintética e capacidade para expressão do CAM nessa porção comparativamente às regiões mediana e a basal. Recentemente vem sendo utilizado um modelo de folhas destacadas mantidas em polietileno glicol (PEG) para a indução do déficit hídrico nos estudos com essa bromélia. A partir disso surgiu a necessidade de se obter informações sobre o efeito da aplicação do PEG em plantas inteiras. Dessa forma, o objetivo deste trabalho visou avaliar o metabolismo fotossintético e caracterizar o perfil espacial de carboidratos não estruturais, levando em consideração a divisão funcional existente na folha dessa espécie, ao longo de um período de restrição hídrica imposta pela aplicação de uma solução de PEG 30%. Foram realizadas medidas de assimilação líquida de carbono, transpiração, condutância estomática, coeficiente de extinção não fotossintético e eficiência potencial/operacional do PSII. Amostras foram coletas para a quantificação do teor relativo de água, clorofilas, carotenoides, malato e carboidratos não estruturais. Com a indução do déficit hídrico ocorreu uma redução do conteúdo hídrico e consequentemente houve a regulação positiva do CAM em todas as porções, com destaque para a região mediana no quarto dia de tratamento. Foi possível observar tanto redução quanto acréscimo na expressão do CAM por meio da variação no acúmulo noturno de malato ao longo dos dias. Na situação controle, o padrão diurno de assimilação líquida de CO2 foi semelhante à de uma espécie C3. Porém, associado a esse padrão, ocorreu uma variação noite/dia no conteúdo de malato, caracterizando-a como um CAM cycling. Ao final do período de déficit hídrico, a assimilação de CO2 atmosférico foi restrita a dois breves momentos no dia, não alcançando completamente o CAM idling. Nesse ponto, a reciclagem do CO2 pela formação do malato apresentou importância na redução da perda de carbono. Por meio de um fluorômetro de imagem foi possível, pela primeira vez para essa espécie, observar regiões de maior eficiência do PSII que podem estar relacionadas à disponibilidade de CO2 proveniente da descarboxilação do malato. Quanto à avaliação dos carboidratos não estruturais, o amido apresentou comportamento inverso ao do malato, possivelmente sendo o carboidrato utilizado na formação de fosfoenolpiruvato. Foi observada uma maior quantidade de amido no conjunto ápice/mediana sugerindo uma fonte enquanto que, um maior conteúdo de açúcares solúveis na base sugere um dreno de utilização / Abstract: Epiphytic species are subjected to periods of water shortage resulting from seasonal rainfall and the lack access to ground water. However, some species are capable of inducing Crassulacean Acid Metabolism (CAM) in order to maintain the water content. Guzmania monostachia is an epiphytic bromeliad capable of CAM up-regulating when exposed to water deficit. In a previous study, this species exhibited CAM idling when submitted to drought during seven days. This type of CAM is characterized by closure stomata during day/night and a nocturnal acidification. Additionally, leaf functional gradient was observed: the apex portion present higher stomata density, amount of photosynthetic pigment, and PEPC/MDH activities when compared to the middle and basal portions. Recently, a model with detached leaves maintained in a polyethylene glycol (PEG) solution to induce water deficit has been used in several studies in this bromeliad. Therefore, further information was needed to understand the effect of use PEG in whole plants. Based on those results, this study aimed to evaluate the photosynthetic metabolism and the non-structural carbohydrate spatial distribution along the leaf in this species under drought condition induced by PEG 30%. Measurements of gas exchange, transpiration, stomatal conductance, non-photochemical quenching and potential/operational efficiency of PSII were performed and samples were collected to measure relative water content, chlorophylls, carotenoids, malate and non-structural carbohydrate amounts. In the fourth day of water deficit, the reduction of relative water content resulted in up-regulation of CAM mainly in the middle portion. It was possible to observe up and down-regulation of CAM by variation in nocturnal accumulation of malate along the days. In the control, the pattern of net carbon assimilation was similar to C3 species. However, this pattern was accompanied by variation in dawn/dusk malate, characterizing it as a CAM cycling. At the end of the drought period, the atmospheric CO2 assimilation was restricted to two short periods of the day, without expression of CAM idling. Nevertheless, CO2 recycling for malate formation showed great relevance to decrease carbon loss. For the first time in this species, it was possible to show spots of high efficiency of PSII (using an imaging fluorometer) suggesting a CO2 availability from malate decarboxylation. Among the non-structural carbohydrates analyzed, only starch presented an inverse diel pattern in relation to malate, indicating that this carbohydrate was possibly used to generate phosphoenolpyruvate. The results suggested that apical and middle portions of the leaf would act as a source due to the high starch levels, while the basal portion would be an utilization sink because of elevated sugar amounts / Mestrado / Biologia Vegetal / Mestre em Biologia Vegetal

Etant donné le rôle des sucres dans de nombreux processus physiopatholologiques importants, le développement de nouveaux outils chimiques pour la glycobiologie est primordial. Une nouvelle approche, consistant à fonctionnaliser sélectivement la liaison C-H anomérique des sucres par insertion d’un métallocarbène, a été développée, offrant ainsi un accès aux α- et β-cétopyranosides. Celle-ci est basée sur l’utilisation d’un bromoacétate ancré en position 2 du sucre, qui joue un rôle clé au sein d’une séquence réactionnelle glycosylation stéréosélective/diazotransfert/fonctionnalisation. En effet, ce bromoacétate permet de contrôler la stéréosélectivité de la réaction de glycosylation par assistance anchimérique ; puis, il permet l’installation du précurseur de carbène afin de favoriser la quaternarisation de la position anomérique. La validation de l’approche sur des méthyl-glycosides modèles a ensuite permis son application à des disaccharides. Ces études ont par ailleurs mis en évidence la tolérance de la transformation vis-à-vis de plusieurs groupements protecteurs communs en chimie des sucres. Enfin, une étude mécanistique, effectuée sur des substrats deutérés, puis complétée par des calculs DFT, a mis en évidence que l’étape de fonctionnalisation impliquait un mécanisme concerté ou dissociatif selon l’orientation équatoriale ou axiale de la liaison C-H anomérique respectivement. / Due to the major roles of carbohydrates in numerous physiopathological processes, it is essential to develop new chemical tools for glycobiology. A new approach, consisting in the selective functionalization of the anomeric C-H bond of carbohydrates by insertion of a metal-carbene, has been developed, thus offering a new access toward α- and β-ketopyranosides. This is based on the use of a key bromoacetate grafted at position 2 of the sugar, which is the corner stone of a stereoselective glycosylation/diazotransfer/functionalization sequence. Indeed, this group firstly controls the stereoselctivity of the glycosylation step by anchimeric assistance; then, it allows the convenient installation of the carbene precursor to promote quaternarization of the anomeric position. The validation of the concept on model methyl-glycosides has allowed its application on disaccharides. In addition, the sequence has shown good tolerance toward many protecting groups commonly used in carbohydrate chemistry. On the basis of mechanistic studies involving deuterium labeled substrates and DFT calculations, we have shown that this functionalization step follows a concerted or stepwise process depending on the equatorial or axial orientation of the anomeric C-H bond, respectively.

L’arboriculture en milieu méditerranéen nécessite un apport d’eau via l’irrigation important, notamment pendant la période estivale. Pour améliorer l’efficience d’utilisation de l’eau, il convient de mieux connaître les besoins en eau de l’arbre et les zones d’exploration et d’exploitation des racines. La croissance des racines varie dans le temps et dans l’espace en lien avec des facteurs endogènes, en particulier la disponibilité en assimilats carbonés, et des facteurs exogènes comme les propriétés du sol. Ces facteurs sont modulés par les pratiques culturales, et en particulier l’irrigation, le compactage du sol et l’éclaircissage, qui affectent la croissance racinaire, et d’une manière générale le fonctionnement global de l’arbre. La dynamique de croissance des racines est ponctuée par deux périodes de croissance intense. La première période de croissance intense se situe tôt dans la saison, en avril-mai, pendant la phase de durcissement du noyau des fruits. En début de saison la demande en carbone à l’échelle de l’arbre est importante (forte croissance des feuilles, fruits, rameaux, racines) et nécessite la mobilisation intense des réserves carbonées présentent sous forme d’amidon. La charge en fruits affecte la croissance des racines et des fruits, et la restriction hydrique affecte surtout la croissance de la partie aérienne. La deuxième période de croissance racinaire intense intervient après la récolte en juillet-août, quand les feuilles et les rameaux ont finis leur croissance. La compétition est moins forte et l’amidon s’accumule, surtout chez les arbres sous restriction hydrique. L’accumulation d’amidon résulte d’une plus grande sensibilité de la croissance au déficit hydrique que la photosynthèse. Pendant les périodes de croissances racinaires intenses, le diamètre apical et la longueur de leur zone apicale non ramifiée des racines sont augmentés, ainsi que les teneurs en sucres solubles dans les pointes racinaires. Les paramètres architecturaux racinaires et les teneurs en sucres solubles dans les pointes racinaires sont de bons indicateurs de la dynamique de croissance racinaire. La répartition des racines au verger est très variable et dépend des propriétés du sol. Les racines des arbres bien alimentés en eau ont colonisé surtout les volumes de sol sous le rang des arbres (proches des goutteurs) jusqu’à 1 m de profondeur. Au contraire les racines des arbres sous-alimentés en eau ont colonisé surtout les 50 cm en profondeur sous le rang et vers l’inter-rang. Les racines se sont réparties dans les zones les moins contraignantes pour leur croissance, qui sont plus restreintes quand l’irrigation est restrictive. En conditions non contraignantes, de par une faible densité de sol ou une forte teneur en eau, la vitesse de croissance, et notamment des grosses racines, est forte. Par contre, les fines racines ont une vitesse de pénétration des sols contraignants plus rapide. La contrainte mécanique entraîne aussi une baisse de la hiérarchisation des systèmes racinaires, les racines latérales seront davantage ramifiées / Fruit tree production in Mediterranean climate needs water supply via irrigation, particularly during summer. To improve water use efficiency, it is necessary to better understand the water need of tree and to localize soil volumes colonized by roots. Root growth varies in time and space following endogenous factors, like carbohydrate availability, and exogenous factors like soil properties. These factors are affected by cultural practices and particularly by irrigation, soil compaction and thinning. Root growth dynamics is marked by two intensive root growth periods. The first period of intensive root growth occurs early in the season, in April-May, during the phase of stone hardening of fruit. Early in the season, carbohydrates demand at tree scale is high (leaves, fruits, shoots, roots are growing) and leads to an intense mobilization of starch reserve. Crop load affects root and fruit growth, and the deficit irrigation affects principally the aerial growth. The second period of intensive root growth occurs after the fruit harvest in July-August, when leaves and shoot stopped their growth. Competition for carbohydrates is reduced and starch contents rise, particularly in trees submitted to deficit irrigation. Starch accumulation traduces a higher sensibility of growth than photosynthesis under deficit irrigation. During intensive root growth periods, apical diameter and the length of apical unbranched zone are increased, and water soluble carbohydrates contents in root tips too. Root architectural parameters and water soluble carbohydrates contents are good indicators of root growth dynamics. Root distribution in orchard is variable and depends of soil properties. Roots of wellirrigated trees colonized especially soil volumes in the row (under drippers) until 1 m in depth. On the opposite, root of trees submitted to deficit irrigation colonized the first 50 cm in depth, both in the row and the inter-row. Root colonized soil volumes where soil mechanical resistance was the lowest, and these soil volumes are more reduced in deficit irrigation. Low soil mechanical resistance, with low bulk density and high water content, allows a higher root elongation and particularly for thick roots. On the opposite, fine roots have a higher elongation in strength soil than thick roots. The soil mechanical resistance leads to reduce the root system hierarchy; lateral roots were more branched

Carbohydrates are ubiquitous components in nature involved in a range of tasks. They cover every cell and contribute both structural stability as well as identity. Lipopolysaccharides are the outermost exposed part of the bacterial cell wall and the primary target for host-pathogen recognition. Understanding the structure and biosynthesis of these polysaccharides is crucial to combat disease and develop new medicine. Structural determinations can be carried out using NMR spectroscopy, a powerful tool giving information on an atomistic scale. This thesis is focused on method development to study polysaccharide structures as well as application on bacterial lipopolysaccharides. The focus has been to incorporate a bioinformatics approach prior to analysis by NMR spectroscopy, and then computer assisted methods to aid in the subsequent analysis of the spectra. The third chapter deals with the recent developments of ECODAB, a tool that can help predict structural fragments in Escherichia coli O-antigens. It was migrated to a relational database and the aforementioned predictions can now be made automatically by ECODAB. The fourth chapter gives insight into the program CASPER, a computer program that helps with structure determination of oligo- and polysaccharides. An approach to determine substituent positions in polysaccharides was investigated. The underlying database was also expanded and the improved capabilities were demonstrated by determining O-antigenic structures that could not previously be solved. The fifth chapter is an application to O‑antigen structures of E. coli strains. This is done by a combination of NMR spectroscopy and bioinformatics to predict components as well as linkages prior to spectra analysis. In the first case, a full structure elucidation was performed on E. coli serogroup O63, and in the second case a demonstration of the bioinformatics approach is done to E. coli serogroup O93. In the sixth chapter, a new version of the CarbBuilder software is presented. This includes a more robust building algorithm that helps build sterically crowded polysaccharide structures, as well as a general expansion of possible components. / <p>At the time of the doctoral defense, the following papers were unpublished and had a status as follows: Paper 3: Manuscript. Paper 5: Manuscript.</p>

Abstract Natural seasonal variation and the effects of cold treatment and nitrogen fertilization on protein expression with special emphasis on dehydrin proteins, were studied using different aged Scots pine (Pinus sylvestris L.). Several different dehydrins were found and their expression depended on the tissue type, tree age or specific treatment. Their concentrations fluctuated seasonally and in response to nitrogen fertilization, but no effects of low temperature on the dehydrins of seedlings were observed. A 60-kDa dehydrin was associated with cold acclimation in the bud and bark tissues of mature trees and in the needles of seedlings. In the needles of mature trees, this dehydrin was associated with springtime desiccation, which was detected as a significant decrease in the osmotic potential of needles. The quantity and quality of soluble proteins altered seasonally in Scots pine tissues, but low temperature treatment alone did not have any effect on the proteins. Soluble protein concentration increased during autumn and decreased in spring in buds and bark, but not in the needles of mature trees. In needles of seedlings, however, protein concentrations altered seasonally. Several proteins, of varying molecular weights, were more abundant in winter in all the tissues studied and some increased in concentration in the nitrogen-fertilized seedlings. The role of these proteins as a storage reserve in Scots pine is discussed. The osmotic potential of needles showed seasonal fluctuation, being high in the summer and low during the winter. Low temperature treatment decreased the osmotic and water potential of needles and increased the concentrations of soluble sugars in seedlings. Based on carbohydrate analyses, the metabolism of seedlings acclimated to low temperature in less than ten days. Nitrogen fertilization increased the content of total nitrogen and the soluble protein concentrations in the needles of seedlings and the growth both in the mature trees and seedlings. Although the frost resistance showed no response to nitrogen-fertilization, the soluble proteins and dehydrins were affected in a manner that suggested an earlier growth resumption of spring in the fertilized trees.

À cause du renforcement des réglementations et de l'évolution de l'opinion publique, les tensioactifs issus de la biomasse sont de plus en plus étudiés. À l'heure actuelle, la diversité structurelle de ces composés reste faible et est bien loin de concurrencer les produits issus des ressources fossiles. Pour enrichir les structures disponibles, nous nous sommes intéressés aux composés furaniques, comme le 2,5-diformylfurane (DFF), synthétisés à partir des sucres. Dans cette optique, nous avons tout d'abord généralisé et optimisé une méthode de synthèse originale du DFF. Ce procédé permet de s'affranchir de l'usage des métaux de transition ou des liquides ioniques. Le système utilisé pour convertir des substrats complexes, comme les amidons, repose sur l'action combinée d'acide borique, de bromure de sodium et d'acide formique dans le diméthylsulfoxyde (DMSO). L'acide borique joue un rôle majeur dans ce système puisqu'il est impliqué dans les étapes de déconstruction de la biomasse, d'isomérisation des unités glucose en fructose et dans la formation du HMF. Le bromure de sodium dans le DMSO permet l'oxydation sélective en DFF alors que l'acide formique accélère cette étape. Dans ces conditions, nous avons été capables de produire du DFF à une échelle de plusieurs dizaines de grammes avec des rendements de l'ordre de 20 %. Après l'optimisation de la synthèse, nous avons également étudié le mécanisme d'oxydation impliqué dans cette transformation. La seconde partie de l'étude a été la transformation du DFF en composés tensioactifs. Au terme d'une synthèse de cinq étapes, nous avons obtenu des produits zwitterioniques originaux avec des rendements d'environ 50 %. Ces composés de type gemini issus de la transformation des sucres montrent de bonnes propriétés comme des concentrations micellaires critiques très faibles comparables aux autres tensioactifs gemini déjà étudiés. / Due to stricter reglementations and the development of public opinion, surfactants synthesized from biomass are becoming increasingly interesting. However, the structural diversity of these compounds is still too low to compete with the products synthesized from fossil resources. In order to increase the diversity of available structures, we focused on common furanic building blocks, such as 2,5-diformylfuran (DFF), which are synthesized form carbohydrates. Firstly, we optimized a new and innovative synthesis of DFF. This method allowed us to use neither transition metal catalysts nor ionic liquids. The system we used to convert complex substrates, such as starch, relies on boric acid, sodium bromide and formic acid in dimethylsulfoxide (DMSO). Boric acid plays a key role in this system. It is involved in the biomass deconstruction, glucose to fructose isomerization and HMF formation. Sodium bromide in DMSO promotes the oxidation to DFF and formic acid increases the rate of this step. By using this system, we were able to synthesized DFF from glucose at multigram scale with 20% yield. After optimization of the synthesis, we investigated the oxidation mechanism. The second part of this work was focused on the conversion of DFF into new gemini surfactants. At the end of a five steps transformation, zwitterionic products were obtained with 50% yield. These gemini type compounds shows interesting properties such as very low critical micellar concentration which are similar to these measured for already reported gemini surfactants.

There exist a wide variety of metabolic responses to different types of carbohydrates and their influence on metabolism during endurance training. Recent studies revealed that the physiological responses to food are far more complex than was previously appreciated. The rapid release of insulin and the decline in blood sugar levels during the first stages of endurance training are linked to the Glycemic Index of foods. Researchers cannot still make use of the old distinction between starchy and sugary food or simple and complex carbohydrates. These distinctions are based on the chemical analysis of the food, which does not totally reflect the effects of these foods on the body. The Glycemic Index is a more reliable guideline to apply in nutritional management for endurance athletes. The major object of the study was to indicate the importance of utilizing the Glycemic Index (GI) as part of the nutritional preparation for endurance events. The study investigated the advantages of ingesting a Low Glycemic Index meal prior to exercise and compared it with the ingestion of a High Glycemic Index meal. A pretest-posttest design was used. Twelve healthy, male and female cyclists participated in the study. Subjects were selected according to their level of training. The total test period consisted of 14 days, which included two different dietary interventions of 7 days each. Diet -and training analysis were done on the subjects prior to the commencement of the study. Each subject completed three exercise trials. The first exercise trial consisted of a V02max test until exhaustion. Two submaximal trials (65 - 70 % of V02max) followed and were preceded by two dietary interventions. The dietary interventions (7 days each) had the same amount of CHO, fat and protein but differed in the Glycemic Index of the pre-exercise meals. The first pre-exercise meal was a High Glycemic Index (HGI) meal. The second pre-exercise meal was a Low Glycemic Index (LGI) meal. The results of the study indicated the advantages of ingesting a Low Glycemic Index meal prior to endurance exercise. The drop in blood glucose levels significantly differed (p<0.05) with an average of 0.68 mmol/L between the two tests after 10 minutes of cycling. It took 20 minutes for the blood sugar level of the first testto reach the same level of the blood sugar level of the second test. After the ingestion of the High Glycemic Index meal in Test 1, the blood lactate levels were significantly higher (p<0.05) during the first 15 minutes. The total distance covered by the subjects was 22.86 km after the first dietary intervention (High glycemic Index food) and 27.43 km after the second dietary intervention (Low glycemic Index food) although it is not statistically significant due to the small sample size. The difference in the distance covered of the two tests is 4.57 km in a period of 50 minutes. Subjects indicated that they experienced more physical strain (higher RPE values) in Test 1 (High Glycemic Index food) than in Test 2 (Low Glycemic Index food) (p<0.05). The study results support the fact that Low glycemic index food may confer an advantage when eaten prior to prolonged strenuous exercise by providing a slow¬releasing source of glucose to the blood without causing extensive hypoglycemia. Proper preparation and the correct choice of the pre-exercise meal can exclude the occurrence of sudden drops in the blood sugar levels. The Glycemic Index can also be successfully applied during and after events to improve performance. / Dissertation (MA (MHS))--University of Pretoria, 2007. / Arts, Languages and Human Movement Studies Education / unrestricted

This dissertation describes the purification and partial characterization of CBP from the epidermis of the monarch butterfly larvae (Danaus plexippus). A yellow protein-carotenoid complex was extracted from the yellow pigmented epidermal tissue from monarch butterfly larvae by homogenization. Additional steps in the purification process included differential precipitation with ammonium sulfate, cation and anion chromatography, and lastly size exclusion chromatography. Polyacrylamide gel electrophoresis demonstrates that a single protein was isolated (M-LBP) having a ~60 kDa molecular weight, the value has subsequently been confirmed by HR-tandem MS. Lutein is the sole carotenoid bound by M-LBP with a stoichiometry of the binding of 2: 1. Immunohistochemistry results show that M-LBP has no cross-reactivity to antibodies for silk worm CBP (Bombix mori) but does have cross-reactivity with antibodies for horn worm epidermal CBP (Agrius convolvuli). Binding affinities were determined using surface plasmon resonance for the carotenoids lutein (KD = 18.6 ± 0.7), R,R-zeaxanthin (KD = 990 ± 60), R,S-zeaxanthin (KD = 60 ± 2). Tryptophyphan fluorescence lifetimes were determined for the apoprotein and compared to those of the native M-LBP. Tryptophan fluorescence lifetimes were found to be 3.9 ns and 3.0 ns, respectively for these two forms of the protein, indicating that upon dissociation of the carotenoid from the protein the tryptophan fluorophore adopts a position where it has less interaction with the polar surface environment.

Os artrópodes constituem as formas mais abundantes de vida animal no planeta. O sistema digestivo dos Arthropoda é uma das mais importantes superfícies de contato com o ambiente. Pouco se conhece a respeito do sistema digestivo dos aracnídeos. Identificamos as principais carboidrases digestivas em três Arachnida modelo: Tityus serrulatus, Amblyomma cajennense e Nephilengys cruentata. As &#945;-fucosidases são glicosídeo hidrolases que catalisam a hidrólise de ligações glicosídicas entre uma fucose ligada a outro carboidrato ou outro tipo de molécula. Estas enzimas são fundamentais no processamento de glicoproteínas e glicolipídeos. Observamos altas atividades de &#945;-fucosidases em todas as espécies estudadas e estas enzimas foram isoladas e caracterizadas. / Arthropods are the most abundant animal life form on the planet. The digestive system of Arthropoda is one of the most important areas of contact with the environment. The study of Arachnida digestive enzymes has been sporadic and remains a largely unexplored area. We identified the most important digestive carbohydrases in three Arachnida models: Tityus serrulatus, Amblyomma cajennense and Nephilengys cruentata. The &#945;-fucosidases are glycoside hydrolases that catalyze the hydrolysis of glycosidic links between a fucose linked to other carbohydrate or another type of molecule. These enzymes are essential in the processing of glycoprotein and glycolipids. We showed that all models tested presented high activities of digestive &#945;-fucosidases and these enzymes were isolated and characterized.