O pedestrianismo vem se tornando cada vez mais expressivo na atualidade, principalmente por assegurar condições de bem-estar físico, psicológico e social ao praticante. Partindo-se do pressuposto que deficiências nutricionais prejudicam o desempenho durante a corrida, corredores de longa duração vêm usufruindo da suplementação esportiva com o intuito de otimizar o rendimento. Todavia, faltam evidências científicas quanto aos recursos ergogênicos proporcionados pelos diversos suplementos alimentares comercializados. Sendo assim, o objetivo desta pesquisa foi comparar o desempenho de corredores de longa duração em um protocolo de exercício físico em esteira composto por três situações diferentes de suplementação: placebo, carboidrato e carboidrato associado à proteína hidrolisada. A amostra foi composta por 14 atletas do sexo masculino e o estudo apresentou design cross over com caráter duplo-cego. Inicialmente, foi realizada avaliação cardiorrespiratória e nutricional, sendo que esta última foi composta por antropometria (peso e estatura) e composição corporal (dobras cutâneas). Os indivíduos foram submetidos a um exercício físico em esteira constituído por 60 minutos com velocidade e inclinação correspondente ao Limiar Anaeróbio e posteriormente perduraram a atividade até a exaustão com intensidade 10% acima desta variável. Foi analisado o consumo alimentar por meio drae feinrgeenstetã ao energética diária e da quantidade de macronutrientes ingeridos nas 24 e 2 horas antecedentes aos exercícios. Mensurou-se a glicemia, o lactato sanguíneo, a percepção subjetiva de esforço e a frequência cardíaca durante os três protocolos de suplementação. A glicemia inicial e 5 minutos após a exaustão foi menor para o grupo placebo quando comparada aos outros dois grupos. Houve um aumento na glicemia e no lactato nos três grupos ao comparar momentos antes e durante o exercício (60 minutos) com momentos após o término da atividade (exaustão e 5 minutos após este estágio). Observou-se aumento na percepção subjetiva de esforço e na frequência cardíaca nos três grupos concomitantemente ao aumento da duração e/ou intensidade do exercício. Ao analisar o consumo alimentar e o tempo de exaustão dos atletas não foram constatadas diferenças estatísticas entre os três suplementos utilizados. Diante disto, analisando agudamente, não há justificativa para adicionar proteína ao suplemento de carboidrato durante a realização de exercícios de endurance com características semelhantes ao de nosso estudo. / The pedestrianism is becoming increasingly significant nowadays, mainly for ensuring conditions of physical, psychological and social well-being practitioner. Starting from the assumption that nutritional deficiencies impair performance during the race, long runners come enjoying the sports supplementation in order to optimize performance. However, they lack scientific evidence about the ergogenic resources provided by the various food supplements marketed. Thus, the aim of this study was to compare the performance of long runners in a physical exercise protocol on a treadmill consists of three different situations supplementation: placebo, carbohydrate and carbohydrate associated with hydrolyzed protein. The sample consisted of 14 male athletes and this study was double-blind with crossover design. Initially, individual performed cardiopulmonary and nutritional evaluation and the latter consisted of anthropometry (weight and height) and body composition (skinfold). The subjects underwent a physical exercise on a treadmill consists of 60 minutes with speed and inclination corresponding to Anaerobic Threshold and then endured the activity to exhaustion intensity with respect to 10% higher this variable. Food consumption through daily energy intake and the amount of macronutrients ingested in 24 and 02 hours before the exercise was analyzed. blood glucose, blood lactate, the perceived exertion and heart rat eI td wurainsg m theea stuhrreede supplementary protocols. The initial blood glucose and 5 minutes after the exhaustion was lower in the placebo group compared to the other two groups. There was an increase in blood glucose and lactate in the three groups when comparing time before and during exercise (60 minutes) with moments after the end of the activity (exhaustion and 5 minutes after this stage). There was an increase in perceived exertion and heart rate in the three groups concomitantly with increasing duration and/or intensity of exercise. By analyzing the food consumption and time to exhaustion of athletes were not found statistical differences between the three supplements used. Given this, there would be no acute justification for adding protein to carbohydrate supplement when performing endurance exercise with characteristics similar to our study.

A sacarose é o dissacarídeo mais abundante na natureza, encontrado na forma pura. É formada pela ligação da hidroxila do C1 da α-D-glicose com a hidroxila do C2 da β-D-frutose. Sua hidrólise em meio ácido produz 1 mol de glicose e 1 mol de frutose. O mecanismo envolve a presença de um intermediário, que corresponde à etapa lenta da reação de hidrólise. A sacarose foi hidrolisada em meio neutro, utilizando-se uma mistura de ésteres derivados do óleo de mamona, produzida pelo Laboratório de Química Analítica e Tecnologia de Polímeros (GQATP). O mecanismo da hidrólise em meio neutro, se assemelhou com o mecanismo da hidrólise ácida, apresentando também um complexo aquoso, que determinou a velocidade da reação. Uma solução de sacarose 0,1 mol L-1 foi hidrolisada pela mistura de ésteres dando como produtos finais 0,019 mol L-1 de glicose e 0,017 mol L-1 de frutose. Verificou-se que a produção dos monossacarídeos foi possível depois do oitavo dia de reação. Soluções de sacarose com diferentes concentrações de cloreto de sódio (NaCl) também foram hidrolisadas, uma vez que este estudo foi importante para se observar a influência da força iônica na velocidade da reação de decomposição. Uma solução de sacarose 10 -3 mol L-1, que apresenta força iônica igual a 5,0 x 10 -4 mol L-1, ao reagir com a mistura de ésteres, apresentou uma quantidade igual a 9,99 x 10 -4 mol L-1 de açúcares redutores formados, ou seja, 99% de sacarose foi hidrolisada. Mesmo com uma maior velocidade de reação, ainda houve a formação de um complexo aquoso, pois a reação não foi instantânea. Com valores de forças iônicas maiores, houve uma menor produção de glicose e frutose. Já a celulose é um polissacarídeo composto de moléculas de D-glicose, unidas por ligações glicosídicas β-1,4, sendo o principal material estrutural das plantas. O mecanismo da hidrólise da celulose em meio ácido assemelha-se à hidrólise ácida da sacarose, em que, antes de se chegar aos produtos finais da reação, ocorre a formação de um complexo intermediário. A celulose também foi hidrolisada em meio neutro utilizando-se a mistura de ésteres, e foi possível observar a presença de um complexo aquoso, antes de se chegar aos produtos finais de sua decomposição, ou seja, celobiose e/ou glicose. Partindo-se de uma solução 0,2 g L-1 de celulose, chegou-se a formação de 0,14 g L-1 de açúcares redutores. Isso corresponde a 70% de celulose hidrolisada. Com relação ao estudo cinético de decomposição da celulose, comprova-se que se trata de uma reação de segunda ordem, e que o tempo de meia-vida da celulose na mistura de ésteres é de 15,29 dias. Como não se sabe a proporção da mistura de ésteres que reage com a celulose, tanto o tempo de meia-vida quanto a lei de velocidade, foram calculadas somente com relação à celulose. / Sucrose is the most abundant disaccharide in nature, found in pure form. It is formed by binding the C1 hydroxyl of the α-D-glucose with the hydroxyl of the C2 of β-D-fructose. Hydrolysis in acid produces 1 mol of glucose and 1 mol of fructose. The mechanism involves the presence of an intermediary, which is the slow step of the hydrolysis reaction. Sucrose was hydrolyzed in neutral, using a mixture of esters derived from castor oil, produced by the Laboratory of Analytical Chemistry and Technology of Polymers (GQATP). The mechanism of hydrolysis in neutral medium, resembled the mechanism of acid hydrolysis, and also provides an aqueous complex, which determined the rate of reaction. A solution of sucrose 0,1 mol L-1 was hydrolyzed by the mixture of esters giving as final products 0,019 mol L-1 glucose and 0,017 mol L-1 of fructose. It was found that the production of monosaccharides was possible after the eighth day of reaction. Sucrose solutions with different concentrations of sodium chloride (NaCl) were also hydrolysed, since this study was important to observe the influence of ionic strength on reaction rate of decomposition. A sucrose solution 10-3 mol L-1, which represents the ionic strength equal to 5,0 x 10-4 mol L-1 and reacted with a mixture of esters, showed a count of 9,99 x 10-4 mol L-1 of reducing sugars formed, ie, 99% sucrose was hydrolyzed. Even with a higher speed of reaction, there was still the formation of an aqueous complex, because the reaction was not instantaneous. With values higher ionic strengths, there was a lower production of glucose and fructose. Since cellulose is a polysaccharide composed of molecules of D-glucose, linked by glycosidic β-1,4, and the main structural material of plants. The mechanism of hydrolysis of cellulose in an acid similar to the acid hydrolysis of sucrose, in which, before reaching the final products of the reaction causes the formation of an intermediate complex. Cellulose was also hydrolyzed in neutral solution using a mixture of esters, and we observed the presence of an aqueous complex, before we get to the final products of decomposition, ie, cellobiose and/or glucose. Starting from a solution of 0,2 g L-1 of cellulose, it was the formation of 0,14 g L-1 of sugars. This corresponds to 70% of hydrolyzed cellulose. Regarding the kinetics of decomposition of cellulose, we find that it is a second-order reaction, and that the half-life of the mixture of cellulose esters is 15,29 days. As we do not know the proportion of mixed esters which reacts with the cellulose, both the half-life as the rate law, were calculated only with respect to cellulose.

Energy usage is continually on the rise and significant efforts are being extended to provide more renewable energy. One area of exploration is the development of fuel cells, which includes biofuel cells that can extract energy from carbohydrates obtained from biomass. Recently, viologen catalysts have been shown to enhance reaction rates of energy extraction and improve carbohydrate conversion efficiencies. However, characterizing the effects of process parameters such as pH, reactant concentrations, and carbohydrate exposure time to buffer solutions with a rigorous model is lacking. This thesis characterizes the homogeneous reaction between carbohydrates and a methyl viologen catalyst to provide insights on ways to enhance the reaction rates to produce more energy. Specifically, the rate of formation of reduced methyl viologen (MV+) in the presence of carbohydrates was measured based on changes in the MV2+ concentration, carbohydrate concentration, pH, and carbohydrate exposure time. A rigorous mechanistic model of the reaction rate was developed and showed a first-order dependence on OH- concentration, a zero-order dependence when the MV2+ concentration was >> 0.5 mM, and a 3-fold increase in the reaction rate when glucose was pre-incubated in a pH 12 buffer solution for 100 minutes. The pre-incubation effect had a strong dependence on pH. The mechanistic model agreed well with experimental data. This thesis also addresses the decomposition of viologen catalysts. MV2+ decomposition experiments showed a trend seen previously in literature that the rate of decomposition increases with an increase in MV2+ concentration, OH- concentration, and temperature. The data and mechanistic model suggest second order dependence of both MV2+ and OH- concentrations under conditions in this thesis (MV2+ concentrations of 100-300 mM and OH- concentrations of 0.001 M and 0.01 M). An activation energy was found from MV2+ decomposition to be 145 kJ/mol. MMV+ decomposition was shown to decompose anywhere from 6.2 – 16.1 times slower than MV2+. Therefore, MMV+ decomposes slower and is more stable than MV2+. It was also found that MV2+ is more stable than IPV-Cl and IPV-Br. An analysis was performed to find the recommended operating range for MV2+/glucose biofuel cells under different conditions while ensuring that at least a viable amount of energy could be produced.

The Mitochondrial Pyruvate Carrier (MPC) occupies a central metabolic node by transporting cytosolic pyruvate into the mitochondrial matrix, thereby linking glycolysis with mitochondrial metabolism. Two reported human MPC1 mutations cause developmental abnormalities, neurological problems, metabolic deficits, and for one patient, early death. We aimed to understand biochemical mechanisms by which the human patient c.C289T and c.T236A MPC1 alleles disrupt MPC function. MPC1 c.C289T encodes two protein variants, a mis-spliced, truncation mutant (A58G) and full-length point mutant (R97W). MPC1 c.T236A encodes a full-length point mutant (L79H). Using human patient fibroblasts and complementation of CRISPR-deleted, MPC1 null mouse C2C12 cells, we investigated how MPC1 mutations cause MPC deficiency. Truncated MPC1 A58G protein was intrinsically unstable and failed to form MPC complexes. The MPC1 R97W protein was less stable but when overexpressed formed complexes with MPC2 that retained pyruvate transport activity. Conversely, MPC1 L79H protein formed stable complexes with MPC2, but these complexes failed to transport pyruvate. These findings inform MPC structure-function relationships and delineate three distinct biochemical pathologies resulting from human patient MPC1 mutations and inform fundamental MPC structure-function relationships. These results also demonstrate an efficient molecular genetic system using the mouse C2C12 cell line to mechanistically investigate human inborn errors in pyruvate metabolism.

Carbon flow into developing ovaries has been reported to be important in seed retention and seed size. Invertase, which cleaves sucrose into glucose and fructose has been shown to be important in rapidly expanding tissue, such as early root growth or during tiller expansion. The manipulation of invertase activity with over-the-top applications of agrochemicals may prevent the detrimental effects of abiotic stress by altering the source/sink relationship. These experiments examined economically important tissues in rice production during critical developmental stages under abiotic stress. Field and greenhouse studies were conducted under normal growing conditions using local management practices. Plants were treated with exogenous chemicals that affect the activity of invertase during the early-grain fill stage on field grown plants. Other plants were exposed to elevated nighttime temperature of 30°C for 4 d using a free-air, infrared heating device in the greenhouse. Rice was also treated at mid- to late-grain fill stage of the main crop to identify the impact of the exogenous chemicals on developing ratoon tiller buds. The activity of soluble acid invertase (SAI), concentrations of glucose, fructose, sucrose, and starch were determined in penultimate leaves, panicles, and main-crop stem segments during ratoon tiller bud expansion, using the enzyme-coupled stoichiometric production of NADH measured spectrophotometrically at 340nm. The results suggest SAI, carbohydrates, and agronomic characters are influenced by exogenous chemicals at the applied rates. The thidiazuron treatment caused an unidentified stress event. The stress was confirmed by increased hexose concentration and the proportion of hexose concentration to sucrose concentration. This stress reduced the main-crop grain yield, but not the ratoon yield or total grain yield. An interaction between the ammonium molybdate treatment and high nighttime temperature was seen in the panicle. The ranked difference was reduced by the high nighttime temperature from the ambient nighttime temperature for the SAI activity, TSC content, starch content, and TNC content. The tested chemicals and rates are not appropriate for commercial rice production because the effect of the exogenous chemicals do not appear to consistently aid rice plants to counteract the detrimental effects of abiotic stress.

The Lattrell-Dax method of nitrite-mediated substitution of carbohydrate triflates is an efficient method to generate structures of inverse configuration. In this study, the effects of the neighboring group on the Lattrell-Dax inversion were explored. A new carbohydrate/anion host-guest system was discovered and the ambident reactivity of the nitrite anion was found to cause a complicated behavior of the reaction. It has been demonstrated that a neighboring equatorial ester group plays a highly important role in this carbohydrate epimerization reaction, restricting the nitrite N-attack, thus resulting in O-attack only and inducing the formation of inversion compounds in good yields. Based on this effect, efficient synthetic routes to a range of carbohydrate structures, notably β-D-mannosides and β-D-talosides, were designed by use of double parallel and double serial inversion. A supramolecularly activated, triggered cascade reaction was also developed. This cascade reaction is triggered by a deprotonation process that is activated by anions. It was found that the anions can activate this reaction following their hydrogen bonding tendencies to the hydroxyl group in aprotic solvents. / QC 20100709

The present thesis honors contemporary molecular dynamics simulation methodologies which provide powerful means to predict data, interpret observations and widen our understanding of the dynamics, structures and interactions of carbohydrate systems. With this as starting point my thesis work embarked on several cutting edge problems summarized as follows. In my first work the thermal response in crystal cellulose Iβ was studied with special emphasis on the temperature dependence of the crystal unit cell parameters and the organization of the hydrogen bonding network. The favorable comparison with available experimental data, like the phase transition temperature, the X-ray diffraction crystal structures of cellulose Iβ at room and high temperatures, and temperature dependent IR spectra supported our conclusions on the good performance of the GLYCAM06 force field for the description of cellulose crystals, and that a cautious parameterization of the non-bonded interaction terms in a force field is critical for the correct prediction of the thermal response in cellulose crystals. The adsorption properties of xyloglucans on the cellulose Iβ surface were investigated in my second paper. In our simulations, the interaction energies between xyloglucan and cellulose in water were found to be considerably lower than those in vacuo. The van der Waals interactions played a prevailing role over the electrostatic interactions in the adsorption. Though the variation in one side chain did not have much influence on the interaction energy and the binding affinity, it did affect the structural properties of the adsorbed xyloglucans. The interaction of the tetradecasaccharide XXXGXXXG in complex with the hybrid aspen xyloglucan endo-transglycosylase PttXET16-34 was studied in the third paper. The effect of the charge state of the “nucleophile helper” residue Asp87 on the PttXET16-34 active site structure was emphasized. The results indicate that the catalysis is optimal when the catalytic nucleophile is deprotonated, while the “helper” residue and general acid/base residue are both protonated. In my forth paper, the working mechanism for a redox-responsive bistable [2]rotaxane based on an α-cyclodextrin ring was investigated. The umbrella sampling technique was employed to calculate the free energy profiles for the shuttling motion of the α-cyclodextrin ring between two recognition sites on the dumbbell of the rotaxane. The calculated free energy profiles verified the binding preferences observed experimentally. The driving force for the shuttling movement of the α-cyclodextrin ring was revealed by the analysis of the free energy components. / QC 20110513

Inhibiting the formation of ice is an essential process commercially, industrially, and medically. Compounds that work to stop the formation of ice have historically possessed drawbacks such as toxicity or prohibitively high active concentrations. One class of molecules, ice recrystallization inhibitors, work to reduce the damage caused by the combination of small ice crystals into larger ones. Recent advances made by the Ben lab have identified small molecule carbohydrate analogues that are highly active in the field of ice recrystallization and have potential in the cryopreservation of living tissue. A similar class of molecules, kinetic hydrate inhibitors, work to prevent the formation of another type of ice – gas hydrate. Gas hydrates are formed by the encapsulation of a molecule of a hydrocarbon inside a growing ice crystal. These compounds become problematic in high pressure and low temperature areas where methane is present - such as an oil pipeline. A recent study has highlighted the effects of antifreeze glycoprotein, a biological ice recrystallization inhibitor, in the inhibition of methane clathrates. Connecting these two fields through the synthesis and testing of small molecule ice recrystallization inhibitors in the inhibition of methane hydrates is unprecedented and may lead to a novel class of compounds.

Bacterial biofilms are surface associated colonies that are of considerable concern and interest to industry, medicine and research. They are resistant to antibiotics, their host’s defences and are able to survive under harsh conditions. Biofilm formation in many bacterial strains are dependent on the production of a polysaccharide intercellular adhesion (PIA), a beta-(1-->6)-N-acetylglucosamine polymer. Vaccines derived from biologically isolated PIA have shown efficacy against clinically isolated strains of E. coli and pathogenic strains of S. aureus in animal models. Accordingly, chemically synthesized neoglycoconjugates based on PIA glycosides will be developed to serve as lead compounds for the development of new antibiotics as well as vaccines against biofilm dependent infections. Described in this thesis is a comprehensive study of the synthesis of PIA oligosaccharides and their deacetylated equivalents, the strategy for installing a stable linker on the free reducing oligosaccharide terminus and finally the conjugation to a model carrier protein for the development of potential neoglycoprotein vaccines.

The synthesis of glycoconjugates is of interest in biological and medicinal research. There are numerous approaches to the synthesis of glycosides involving protecting group free methods. This thesis outlines what has been achieved in the field and two novel approaches for O-glycosidation. The first approach involves the use of a toluenesulfonohydrazide glycoside with a purification handle designed for simple glycoside purifications. The butyl 3-O-octyl-D-glucopryanoside was successfully synthesized but did not have the desired property of yielding simple-to-purify glycosides as products. The second approach uses a thiouronium glycosyl donor; a variety of glycosidations using this donor were investigated. The glucosyl thiouronium salt donor was shown to undergo glycosidation effectively with simple alcohols.