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  • About
  • The Global ETD Search service is a free service for researchers to find electronic theses and dissertations. This service is provided by the Networked Digital Library of Theses and Dissertations.
    Our metadata is collected from universities around the world. If you manage a university/consortium/country archive and want to be added, details can be found on the NDLTD website.
111

Glycosyl disulfides: importance, synthesis and application to chemical and biological systems

Ribeiro Morais, Goreti, Falconer, Robert A. 16 November 2020 (has links)
Yes / The disulfide bond plays an important role in the formation and stabilisation of higher order structures of peptides and proteins, while in recent years interest in this functional group has been extended to carbohydrate chemistry. Rarely found in nature, glycosyl disulfides have attracted significant attention as glycomimetics, with wide biological applications including lectin binding, as key components of dynamic libraries to study carbohydrate structures, the study of metabolic and enzymatic studies, and even as potential drug molecules. This interest has been accompanied and fuelled by the continuous development of new methods to construct the disulfide bond at the anomeric centre. Glycosyl disulfides have also been exploited as versatile intermediates in carbohydrate synthesis, particularly as glycosyl donors. This review focuses on the importance of the disulfide bond in glycobiology and in chemistry, evaluating the different methods available to synthesise glycosyl disulfides. Furthermore, we review the role of glycosyl disulfides as intermediates and/or glycosyl donors for the synthesis of neoglycoproteins and oligosaccharides, before finally considering examples of how this important class of carbohydrates have made an impact in biological and therapeutic contexts. / The authors thank the Institute of Cancer Therapeutics (University of Bradford) Doctoral Training Centre for financial support.
112

Analysis of the role of relative nucleotide concentrations on the regulation of carbohydrate in higher plants

Boussiengui-Boussiengui, Gino 12 1900 (has links)
Thesis (PhD (Genetics))--Stellenbosch University, 2010. / ENGLISH ABSTRACT: The current understanding of the regulation of carbohydrate accumulation is still under investigation despite the tremendous work done in this subject. Purine and pyrimidine nucleotides have been implicated in many biochemical processes in plants. Amongst others, they are building blocks for nucleic acid synthesis, an energy source, precursors for the synthesis of primary products such as sucrose, polysaccharides, phospholipids, as well as secondary products. With the aim of placing adenine and uridine nucleotides in the context of sucrose and starch metabolism and carbon partitioning in higher plant, we have investigated the transcripts, enzymes and metabolites in carbohydrate metabolism and both de novo and salvage of purine and pyrimidine nucleotides in both sugarcane and tobacco tissues. For that purpose, adenylate kinase (ADK) and UMP synthase were chosen for silencing and over expression as they are rate limiting steps of de novo adenine and uridine nucleotides biosynthesis, respectively. Sugarcane with repressed ADK activity showed significant increase in both the starch and adenylate pools. Increase in starch content was highly correlated with reduced ADK activity. As a result of decreased ADK activity, the salvage pathway was up regulated via the increased activity of both adenosine kinase (AK) and adenine phosphoribosyl transferase (APRTase) which positively correlated with increase in adenine nucleotide contents. In addition hexose phosphates and ADP glucose, the committed substrate for starch biosynthesis positively correlated with changes in starch content. A high ratio of ATP/ADP was observed in all transgenic lines compared with the untransformed wild type and suggested to favour starch synthesis. Over expression of cytosolic ADK in tobacco demonstrated an expression of the enzyme where 2/3 of the total activity was in the direction of ADP production. As a result of over expression of ADK, starch content increased in all transgenic plants and positively correlated with changes in the activity of ADK. Despite changes in adenine nucleotide content, the salvage pathway was not activated and no significant changes in both AK and APRTase acivities were found between the transgenic and the untransformed plants. Sucrose synthase (SuSy) activity in breakdown direction positively correlated with changes in starch content suggesting a contribution in the starch accumulation in tobacco plants. In addition the ratio of ATP/ADP was low in all transgenic lines compared with the untransformed wild type. This was in line with the higher content in ADP compare to ATP in all transgenic lines and was supported by the over expression of ADK, and predominantly in the direction of ADP production. Repressed UMP synthase in transgenic sugarcane resulted in increases in sucrose, starch and uridinylate. UDP-glucose, hexose phosphates and uridinylate content positively correlated with changes in sucrose content. Transgenic lines had increased sucrose phosphate synthase (SPS) activity and low activity in SuSy, which suggests alteration of carbon flux toward sucrose. As a result of decreased UMP synthase activity, an up regulation of the salvage pathway was observed and predominantly via increased activity of uridine kinase (UK) which positively correlated with changes in the uridinylate pool. In addition to repressed UMP synthase activity, starch content and adenine nucleotides increased in transgenic lines. Tobacco plants transformed with a cytosolic UMP synthase demonstrated an over expression of the enzyme in all transgenic lines. As a result of over expression of UMP synthase, key metabolites were up regulated, amongst them sucrose. Increase in sucrose content positively correlated with both hexoses and hexose phosphates but not the uridinylate pool. SPS activity positively correlated with increase in sucrose content, and accounted for most of the sucrose synthesized in transgenic lines. Despite the increase in the adenylate pool, no significant changes were observed in starch content. The depletion level of UDP-glucose in all transgenic lines was a mere reflection of the higher activity of UDP glucose pyrophosphorylase (UGPase) in the formation of glucose-1-phosphate. In addition, no salvage pathway was up regulated in transgenic lines. / AFRIKAANSE OPSOMMING: Die huidige beskikbare inligting in verband met die reguleering van koolhidraat akkumulasie word steeds ondersoek ten spyte van die groot hoeveelheid navorsing wat reeds in hierdie verband gedoen is. Purien en pirimidien nukleotide speel ‘n rol in baie biochemiese prosesse in plante. Onder andere is hulle boublokke vir nukleïensuur sintese, ‘n energie bron, voorlopers vir die sintese van primêre produkte soos byvoorbeeld sukrose, polisakkariede, fosfolipiede, asook sekondêre produkte. Met die vooruitsig om adenine- en uridiennukleotide in verband te plaas met sukrose en stysel metabolisme en koolstof afskorting in plante, ondersoek ons hier die transkripte, ensieme en metaboliete in koolhidraat metabolisme in beide de novo en berging van purien en pirimidien nukleotide in suikerriet asook tabak weefsel. Vir hierdie doel is adenilaatkinase (ADK) en UMP-sintase gekies vir uitskakeling en ooruitdrukking, juis omdat hulle tempo vermindering stappe van de novo adenine- en uridiennukleotide biosintese is. Suikerriet met onderdrukte ADK aktiwiteit wys betekenisvolle vermeerdering in beide die stysel en adenilaat poele. Verhoging in styselinhoud was hoogs gekorreleerd met verminderde ADK aktiwiteit. As gevolg van ‘n vermindering in ADK aktiwiteit, is die bergingspad opwaards gereguleer via die vermeerdering van beide adenosienkinase (AK) en adenien-fosforibosieltransferase (APRTase) aktiwiteit wat positief korreleer met die vermeerdering in adeniennukleotied-inhoud. Addisioneel word hexosefosfate en ADP-glukose, die toegewysde substraat vir stysel biosintese, positief gekorreleer met veranderinge in styselinhoud. ‘n Hoë verhouding van ATP/ADP was geobserveer in alle transgeniese lyne in vergelyking met die nie-getransformeerde wilde tipe en blyk stysel sintese te begunstig. Ooruitdrukking van sitologiese ADK in tabak demonstreer die uitdrukking van die ensiem waar 2/3 van die totale aktiwiteit in die rigting van ADP produksie was. As ‘n resultaat van ooruitdrukking van ADK, word stysel inhoud vermeerder in alle transgeniese plante en positief gekorreleer met die verandering in die aktiwiteit van ADK. Ten spyte van veranderinge in adeniennukleotide inhoud was die bergingspad nie geaktiveer nie en geen betekenisvolle veranderinge in beide AK en APRTase aktiwiteit was gevind tussen die transgeniese en nie-transgeniese plante nie. Sukrose sintese (SuSy) aktiwiteit tydens afbreking korreleer positief met die veranderinge in stysel inhoud en dui moontlik op ‘n bydrae in die stysel akkumulasie in tabak plante. Verder was die verhouding van ATP/ADP laag in alle transgeniese lyne in vergelyking met die nie-getransformeerde wilde tipe. Hierdie bevinding word ondersteun deur die hoër inhoud in ADP in vergelyking met ATP in alle transgeniese lyne en word verder ondersteun deur die ooruitdrukking van ADK, hoofsaaklik in die rigting van ADP produksie. Onderdrukte UMP-sintase in transgeniese suikerriet lei tot verhogings in sukrose, stysel en uridienilaat. UDP-glukose, hexose-fosfate en uridienilaat inhoud korreleer positief met die verandering in sukrose inhoud. Transgeniese lyne het verhoogde sukrose-fosfaatsintase (SPS) aktiwiteit en lae SuSy aktiwiteit wat dui op ‘n verandering in koolstof vloei in die rigting van sukrose. As gevolg van die afname in UMP-sintese aktiwiteit, word ‘n verhoogde reguleering van die bergingspad gesien, en dít hoofsaaklik via verhoogde aktiwiteit in uridienkinase (UK) wat positief korreleer met veranderinge in die uridienilaat poel. Addisioneel tot die onderdrukking van UMP-sintase was stysel inhoud en adenine- nucleotides in transgeniese lyne verhoog. Tabak plante wat getransformeer is met sitologiese UMP-sintase demonstreer verhoogde uitdrukking van die ensiem in al die transgeniese lyne. As ‘n resultaat van ooruitdrukking van UMP-sintase is sleutel metaboliete, onderandere sucrose, oorgereguleer. ‘n Verhoging in sukrose inhoud korreleer positief met beide hexose en hexose-fosfate maar nie met die uridienilaat poel nie. SPS aktiwiteit korreleer positief met die verhoging in sukrose inhoud en verklaar die meeste van die sukrose vervaardig in transgeniese lyne. Ten spyte van die verhoging in die adenilaat poel word geen noemenswaardige veranderinge gesien in die stysel inhoud nie. Die uitputtingsvlak van die UDP-glukose in alle transgeniese lyne was slegs ‘n aanduiding van die hoër aktiwiteit van UDP-glukose pirofosforilase (UGPase) in die formasie van glukose-1-fosfaat. Verder was geen bergingspad opgereguleer in die transgeniese lyne nie. / The South African Sugarcane Research Institute and the Gabonese Government who provided the financial support for this work
113

Characterization of carbohydrate based vaccines / Caractérisation de vaccin à base glucides

Tontini, Marta 26 October 2012 (has links)
CARACTERISATION DE VACCINS A BASE DE GLUCIDESVariables influençant l'immunogénicité et propriétés physico-chimiques des vaccins glycoconjuguésDe nombreux aspects peuvent influer sur l'immunogénicité des vaccins conjugués et les principales variables étudiées jusqu'ici sont la taille du fragment saccharide et la nature des glycosides: taux de protéine dans le conjugué purifié, la stratégie de conjugaison, nature de l’espaceur et la protéine porteuse.La taille de la partie saccharidique et le ratio de cette partie / protéine a été étudiée dans différents travaux de Seppälä et Mäkelä,.Dans l'une des premières études sur l'effet de la taille sur l'immunogénicité de la protéine conjuguées à des dextrans, il a été montré que des dextrans de faible poids moléculaire conjugué à l’albumine sérique de poluet induit des réponses anti-dextran fortes chez la souris. L’augmentation de la taille du dextran, a abouti à une réduction de l’immunogénicité. Peeters et al. a montré qu’un tétramère synthétique de Hib unité capsulaire polysaccharide, conjuguée à un support protéique, induit chez des souris adultes, des niveaux d'anticorps primates non humains comparables à un conjugué Hib commercial. Ces niveaux sont plus élevées que ceux induits par un trimère répété, ce qui indique que pour le Hib un minimum de huit sucres est nécessaire pour une bonne réponse immunologique. Laferrière et al. a trouvé peu d'influence de la longueur de la chaîne glucidique sur l'immunogénicité des vaccins antipneumococciques conjugués chez la souris. Pozsgay et al. a étudié chez la souris, l'immunogénicité de l’oligosaccharides du LPS de Shigella dysenteriae de type 1 conjugué à l'albumine sérique humaine (HSA). Les auteurs ont constaté que les octa-, dodéca-, et des fragments de hexadécasaccharides induit des niveaux élevés d'anticorps IgG après trois injections. Ces niveaux sont supérieurs à ceux obtenus avec un conjugué tétrasaccharidique. L'influence du ratio glucides / protéine est différente pour les trois conjugués. Le conjugué octasaccharide-HSA avec la plus forte densité provoque une bonne réponse immunitaire, tandis que dans le cas des conjugués dodéca- et hexadécasaccharides, la densité médiane est optimal. Ces études suggèrent que la longueur de la chaîne d'oligosaccharides et le chargement de l’haptène peuvent être liés entre eux pour déterminer l'immunogénicité des vaccins glycoconjugués.L'espaceur est une molécule linéaire courte qui est généralement liée à la chaîne polysaccharidique et la protéine ou de fragments. Il ya des évidences dans la littérature qui suggèrent que les espaceurs rigides, contraints comme le maléimide cyclohexyle, provoquent une importante quantité d'anticorps indésirables, avec le risque de conduire à une réponse immunitaire éloignée de l'épitope ciblé sur la haptène. L'utilisation d'un alkyle souple type maleimido a été rapporté comme un moyen de surmonter l'immunogénicité observée précédemment. Un certain nombre de transporteurs protéiques ont été utilisés jusqu'ici dans l'évaluation préclinique et clinique de vaccins conjugués. Des protéines telles que les anatoxines diphtériques et tétaniques, qui dérivent des toxines respectives, après la décontamination chimique avec le formaldéhyde, ont été initialement choisies comme transporteur en raison de inocuité (tétanos et la vaccination contre la diphtérie). CRM 197, un mutant non toxique de la toxine 61 de la diphtérie a été largement utilisé comme support pour Hib. Un complexe protéique de la membrane externe de méningocoque du sérogroupe B a été utilisé par Merck comme support pour leur vaccin conjugué Hib. GSK dans leur vaccin antipneumococcique, conjugué multivalent, introduit la protéine D Hib liée à la plupart des polysaccharides inclus dans le vaccin. L'équipe de John Robbins fait un large usage de la forme recombinante non toxique de l’exo-toxine de Pseudomonas aeruginosa comme support contre Staphylococcus aureus de type 5 et 8 ainsi que pour Salmonella. / CHARACTERIZATION OF CARBOHYDRATE BASED VACCINES Variables influencing the immunogenicity and physicochemical properties of glycoconjugate vaccinesMany aspects can influence the immunogenicity of conjugate vaccines and the main variables investigated so far are the size of the saccharide moiety, the saccharide:protein ratio in the purified conjugate, the conjugation strategy, the nature of the spacer and the protein carrier. The size of the saccharide moiety and saccharide/protein ratio were investigated in different works such as Seppälä and Mäkelä in one of the first studies on the effect of size and chemistry on the immunogenicity of dextrans-protein conjugates found that dextrans of low molecular weight conjugated to chicken serum albumin, induced strong anti-dextran responses in mice, while increasing the dextrans' size resulted in reduced immunogenicity.47 Peeters et al. showed that a synthetic tetramer of Hib capsular polysaccharide repeating unit, conjugated to a protein carrier, induced in adult mice and non-human primates antibody levels comparable to a commercial Hib conjugate and higher than those induced by a trimer, indicating that for Hib a minimum of eight sugars is needed for a proper immunological response.48 Laferriere et al. found little influence of the carbohydrate chain length on the immunogenicity of pneumococcal conjugate vaccines in mice.49 Pozsgay et al. studied the immunogenicity in mice of synthetic Shigella dysenteriae type 1 LPS oligosaccharides conjugated to human serum albumin (HSA). The authors found that octa-, dodeca-, and hexadecasaccharide fragments induced high levels of lipopolysaccharide binding IgG antibodies in mice after three injections and were superior to a tetrasaccharide conjugate. The influence of the carbohydrate/protein ratio was different for the three conjugates. The octasaccharide-HSA conjugate with the highest density evoked a good immune response, while in the case of dodeca- and hexadecasaccharide conjugates, the median density was optimal.50 These studies suggest that oligosaccharide chain length and hapten loading might be interconnected in determining the immunogenicity of glycoconjugate vaccines. The spacer is a short linear molecule that is generally linked to the polysaccharide chain or to the protein or to both moieties, depending on the chemistry, used to facilitate the coupling between the protein and sugar. There are evidences in the literature which suggest that rigid, constrained spacers like cyclohexyl maleimide, elicit a significant amount of undesirable antibodies, with the risk of driving the immune response away from the targeted epitope on the hapten.51 52 The use of a flexible alkyl type maleimido spacer has been reported as a way to overcome the previous observed immunogenicity of cyclic maleimide linkers.53 A number of protein carriers have been used so far in preclinical and clinical evaluation of conjugate vaccines. 54 55 56 57 58 59 60Proteins such as diphtheria and tetanus toxoids, which derive from the respective toxins after chemical detoxification with formaldehyde, were initially selected as carrier because of the safety track record accumulated with tetanus and diphtheria vaccination. CRM197, a non-toxic mutant of diphtheria toxin61 which instead does not need chemical detoxification, has been extensively used as carrier for licensed Hib, pneumococcal, meningococcal conjugate vaccines and for other vaccines being developed. An outer membrane protein complex of serogroup B meningococcus has been used by Merck as carrier for their Hib conjugate vaccine.62 GSK in their multivalent pneumococcal conjugate vaccine introduced the use of the Hib-related protein D as carrier for most of the polysaccharides included into the vaccine.63 64 The team of John Robbins made extensive use of the recombinant non toxic form of Pseudomonas aeruginosa exo-toxin as carrier for Staphylococcus aureus type 5 and 8 as well as for Salmonella
114

Enzimas do metabolismo de frutanos em Vernonia herbacea (VELL.) Rusby. / Fructan metabolizing enzymes in Vernonia herbacea (VELL.) Rusby.

Asega, Amanda Francine 14 April 2003 (has links)
A ocorrência de frutanos em espécies de Asteraceae foi amplamente documentada para a flora da região de cerrado da Reserva Biológica e Estação Experimental de Moji Guaçu. Dentre estas espécies destaca-se Vernonia herbacea, uma planta perene que apresenta órgãos subterrâneos de reserva, denominados rizóforos, que acumulam altos teores de frutanos do tipo inulina. Seu crescimento sazonal é caracterizado pela brotação das gemas existentes nos rizóforos, na primavera, seguida de floração e crescimento vegetativo intenso no verão e dormência no inverno. O conteúdo de frutanos diminui durante a brotação e floração, pois este carboidrato parece ser utilizado para a regeneração dos ramos aéreos que ocorre nesta fase. Estudos preliminares mostraram que a FEH, responsável pela mobilização dos frutanos, apresenta atividade elevada apenas durante a brotação; a despolimerização dos frutanos nos rizóforos de V. herbacea nesta fase foi evidenciada pelo aumento de açúcar redutor, especialmente frutose. O presente estudo teve como objetivo principal a análise da atividade das enzimas envolvidas no metabolismo de frutanos e análise do conteúdo e da composição de frutanos em rizóforos de V. herbacea induzidas à brotação. A brotação foi induzida pela remoção dos ramos aéreos e as atividades das enzimas FEH , SST, FFT e INV foram determinadas a cada quatro dias durante um mês após a poda. Um aumento na atividade da FEH foi observado entre os dias 13 e 20 após a poda, o que coincidiu com o início da brotação dos novos ramos que ocorreu no 13º dia. Este resultado sugere que a despolimerização de frutanos e a brotação são processos concomitantes em V. herbacea, que ocorrem naturalmente durante o ciclo fenológico, mas que também podem ser induzidos em outras fases do ciclo fenológico pela remoção dos ramos aéreos. A FFT parece atuar junto à FEH na diminuição do tamanho das cadeias de frutanos durante a rebrota, enquanto a SST é inibida devido, possivelmente, à interrupção do fornecimento de sacarose aos rizóforos pelos órgãos aéreos. Para a caracterização e purificação parcial da FEH de rizóforos de V. herbacea foram utilizados rizóforos de plantas induzidas à brotação através da remoção dos ramos aéreos. O extrato bruto apresentou, para atividade de FEH, pH ótimo de 4,5, temperatura ótima em 30 ºC e curva sigmoidal de concentração de substrato, sugerindo tratar-se de uma enzima alostérica. Esta enzima também apresentou maior especificidade por ligações do tipo b-2,1 do que sobre ligações b-2,6, e maior afinidade por frutanos de cadeias curtas quando comparadas com frutanos de cadeias longas. Utilizando precipitação com sulfato de amônia, cromatografia de afinidade e cromatografias de troca aniônica e catiônica, quatro frações com atividade de FEH foram purificadas. Destas quatro frações, duas foram submetidas à cromatografia de exclusão molecular, sendo que os pesos moleculares estimados para uma delas foi de 21 kDa e para outra, esta medida situou-se entre 155 e 39 kDa, devido à ampla faixa de exclusão da coluna utilizada. Os pesos moleculares das outras duas frações foram estimados por SDS-PAGE, sendo que as bandas visualizadas corresponderam a 81,3 e 57,5 kDa para uma e 57,5 kDa para a outra fração. / The occurrence of fructans has been reported in native species of a cerrado area of Reserva Biológica e Estação Experimental de Moji Guaçu. Vernonia herbacea, one of these species, presents underground organs named rhizophores which accumulate fructans of the inulin type as reserve carbohydrate. The seasonal growth pattern exhibited by plants of V. herbacea includes sprouting of buds from the rhizophores in spring, followed by a period of flower development and vegetative growth in summer, and dormancy in winter. The fructan content decreases during sprouting and flowering due to mobilization of this carbohydrate during the regeneration of the new shoots. Preliminary studies showed that FEH, the enzyme responsible for the mobilization of fructan, shows high activity only during sprouting. Fructan mobilization was detected by the increase in the amount of reducing sugar released, namely fructose. The aim of this work was to analyze the activities of the enzymes of fructan metabolism, FEH, SST, FFT and invertase and the fructan contents in rhizophores of plants induced to sprouting by defoliation. The enzyme activities were measured every 4 th day for a month after defoliation. Sprouting of new shoots started around the 13 th day, while an increase in FEH activity was detected between 13 and 20 days after defoliation. The results suggest that fructan depolimerization and sprouting are concomitant processes in V. herbacea that occur naturally during the phenological cycle; however, these processes can also be induced by defoliation during other stages of the phenological cycle. FFT seems to act together with FEH by catalyzing the decrease in fructan chain size during shoot regrowth, while SST was inhibited, possibly, due to interruption of sucrose supply to rhizophores from the aerial organs. The characterization and partial purification of FEH were done using rhizophores from plants which were induced to sprouting by defoliation. The optimal pH and temperature for FEH activity were pH4,5 and 30ºC, respectively. The substrate concentration curve exhibited a sigmoid shape, suggesting that FEH of V. herbacea is an alosteric enzyme. Additionally, this enzyme shows more specificity to b-2,1 than to b-2,6 linked fructan and higher affinity for short chain when compared to long chain fructans. Four fractions presenting FEH activity were partially purified by a combination of ammonium sulfate precipitation, affinity chromatography, and anion and cation exchange chromatographies. Two of these were submitted to size exclusion chromatography and the apparent molecular size for one of them was 21 kDa and for the other it was estimated to be between 39 and 155 kDa, due to the wide exclusion limit of the column used. The molecular size of the next two fractions were estimated by SDS- PAGE and the visualized bands corresponded to 81.3 and 57.5 kDa for the first fraction and to 57.5 kDa for the second one.
115

Efficient carbohydrate synthesis by controlled inversion strategies

Dong, Hai January 2006 (has links)
<p>The Lattrell-Dax method of nitrite-mediated substitution of carbohydrate triflates is an efficient method to generate structures of inverse configuration. In this study it has been demonstrated that a neighboring equatorial ester group plays a highly important role in this carbohydrate epimerization reaction, 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 the ester activation effect for neighboring groups, a double parallel as well as a double serial inversion strategy was developed.</p>
116

Effects of high-carbohydrate and low-fat versus high-protein and low-carbohydrate diets on high-intensity aerobic exercise /

Toma, Kumika. January 2009 (has links)
Thesis (Ph.D.)--Ohio University, August, 2009. / Release of full electronic text on OhioLINK has been delayed until September 1, 2012. Includes bibliographical references (leaves 157-195)
117

Effects of high-carbohydrate and low-fat versus high-protein and low-carbohydrate diets on high-intensity aerobic exercise

Toma, Kumika. January 2009 (has links)
Thesis (Ph.D.)--Ohio University, August, 2009. / Title from PDF t.p. Release of full electronic text on OhioLINK has been delayed until September 1, 2012. Includes bibliographical references (leaves 157-195)
118

The effects of sports drinks containing caffeine and carbohydrate on soccer-specific skill performance during match-induced fatigue

Jacobson, Marc A. 26 April 2011 (has links)
A ninety minute competitive soccer match consists of many intermittent sprints resulting in fatigue, and consequently, a reduction in skill performance. The combination of caffeine and carbohydrate (CHO) has been shown to have ergogenic effects which help maintain skill measures during fatiguing states, however, there has been little research investigating this combination on soccer performance. Therefore, the aim of this study was to examine the effects of three sports drinks, including a placebo (PLA), a 6% CHO drink, and CHO + caffeine (CCAF; 5 mg/Kg body mass (BM)) on soccer-specific skills, throughout a fatigue-inducing soccer match. Twelve male soccer players completed three ninety minute intersquad matches played outdoors on a grass field in a randomized crossover design. Players consumed 5 ml/kg BM 45 minutes prior to kickoff and 3 ml/kg BM every 15 minutes during match play. Soccer passing skill was measured using the Loughborough Soccer Passing Test (LSPT), shot speed, and 20m sprint performance were measured pre-match, immediately at halftime and immediately post-match. Countermovement jump (CMJ) was measured pre-match and post-match. Heart rate (HR) was measured continuously. Blood lactate, rating of perceived exertion (RPE), and perceived fatigue were assessed every fifteen minutes throughout the match. Urine was collected pre-match for analysis of urine specific gravity (USG). BM was measured pre-match and post-match. LSPT total performance time was significantly better in the CCAF trial compared to the PLA trial at halftime (55.3 ± 10.3 s vs 66.5 ±8.7 s, p = .027). There were also significant improvements in penalty time (CCAF 8.2 ± 7.6 s vs. PLA 16.6 ± 7.8 s, p = .042) and movement time (CCAF 8.2 ± 7.6 s vs. PLA 16.6 ± 7.8 s, p = .028) during the CCAF trial in comparison to the PLA at halftime. HR and blood lactate was elevated throughout the PLA trial in comparison to the CHO trial. There were no other significant findings. Most players (50% - 83%) started all three matches in a dehydrated state (USG > 1.020). The CHO trial had significantly lower sweat rates (0.83 ± 0.25 L/hr) than both the PLA trial (1.06 ± 0.26 L/hr, p = .038) and the CCAF trial (1.11 ± 0.19 L/hr, p = .009). The addition of caffeine to a CHO sports drink significantly improved passing performance (quicker completion time and fewer penalties accumulated) over a PLA. All three sports drinks appeared to be equally as effective in preventing deterioration of soccer skill performance during a game situation. This suggests that the total volume of fluid consumed is of greater importance than the type of fluid. Caffeine appeared to have limited ergogenic effects on skill performance without any negative consequences. / Graduate
119

The influence of hypotonic carbohydrate electrolyte solutions on muscle metabolism and exercise capacity with regards to intermittent high intensity shuttle running

Foskett, Andrew January 2003 (has links)
Performance and exercise capacity in prolonged continuous exercise has been shown to be enhanced by the ingestion of dilute (-6%) carbohydrate-electrolyte (CHO-E) solutions during exercise. There is a wealth of literature on the effects of CHO-E solutions on prolonged, continuous exercise however this is not the case for highintensity intermittent exercise. One reason for this is the difficulty of replicating the demands of multiple-sprint sports within the laboratory. The development of the Loughborough Intermittent Shuttle Running Test (LIST), a protocol that simulates the physiological demand of multiple-sprint sports, has allowed for further study in this area. Sports drinks contain a variety of different types of carbohydrate, including synthetic polymers of maltodextrins. The rationale for these is two-fold; firstly there is some evidence to suggest that these glucose polymers of low osmolality may empty faster than isoenergetic glucose solutions especially at dilute (-6%) concentrations. Secondly CHO-E solutions formulated from maltodextrins with low dextrose equivalents are less acidogenic and have lower cariogenic potential than simple glucose solutions, thus making them preferable for tooth health. Therefore the purpose of this thesis was to examine the effects of a 6.4% hypotonic synthetic polymer maltodextrin CHO-E solution on muscle metabolism and physiological performance during prolonged high-intensity intermittent running (LIST). This thesis contains four experimental studies. From the results of study 1 it was concluded that the ingestion of a 6.4% hypotonic CHO-E solution provided no ergogenic benefit to running capacity during the LIST despite blood and hormonal data suggesting that the solution was effective at delivering glucose. As isotonic CHO-E solutions have shown to have ergogenic properties during LIST running the purpose of study 2 was to compare isoenergetic hypotonic and isotonic CHO-E solutions during LIST running. The results from this study suggested that performance, as measured by exercise capacity and sprint speed, was not affected by either treatment. In study 3 the protocol was modified to extend the exercise duration so that a greater demand was placed on the subjects' glycogen reserves. The results from this study further confirmed that exercise capacity in LIST running was not influenced by CHO provision during exercise however the habitual CHO intake of the subjects may have masked any ergogenic properties of the CHO-E solution and led to increased performance in the control trials. In the final study the subjects' pre-trial exercise and dietary CHO intake was manipulated to increase endogenous glycogen concentrations. The results showed that in these subjects the ingestion of a 6.4% hypotonic CHO-E solution increased exercise capacity during the LIST by 21%. Muscle biopsy analyses revealed a lower net muscle glycogen utilisation in the CHO trial post-90 min (p=0.07). There is evidence in the literature to suggest that there is an ergogenic effect of CHO ingestion during the LIST. However the results presented in this thesis suggest that when endogenous glycogen stores are moderate the ingestion of a CHO-E solution may suppress lipid oxidation without a concomitant increase in CHO oxidation and thus LIST capacity is not enhanced. In contrast, when endogenous glycogen concentrations are elevated through dietary manipulation there is an ergogenic benefit from the ingestion of hypotonic CHO-E solutions during LIST exercise.
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Enzimas do metabolismo de frutanos em Vernonia herbacea (VELL.) Rusby. / Fructan metabolizing enzymes in Vernonia herbacea (VELL.) Rusby.

Amanda Francine Asega 14 April 2003 (has links)
A ocorrência de frutanos em espécies de Asteraceae foi amplamente documentada para a flora da região de cerrado da Reserva Biológica e Estação Experimental de Moji Guaçu. Dentre estas espécies destaca-se Vernonia herbacea, uma planta perene que apresenta órgãos subterrâneos de reserva, denominados rizóforos, que acumulam altos teores de frutanos do tipo inulina. Seu crescimento sazonal é caracterizado pela brotação das gemas existentes nos rizóforos, na primavera, seguida de floração e crescimento vegetativo intenso no verão e dormência no inverno. O conteúdo de frutanos diminui durante a brotação e floração, pois este carboidrato parece ser utilizado para a regeneração dos ramos aéreos que ocorre nesta fase. Estudos preliminares mostraram que a FEH, responsável pela mobilização dos frutanos, apresenta atividade elevada apenas durante a brotação; a despolimerização dos frutanos nos rizóforos de V. herbacea nesta fase foi evidenciada pelo aumento de açúcar redutor, especialmente frutose. O presente estudo teve como objetivo principal a análise da atividade das enzimas envolvidas no metabolismo de frutanos e análise do conteúdo e da composição de frutanos em rizóforos de V. herbacea induzidas à brotação. A brotação foi induzida pela remoção dos ramos aéreos e as atividades das enzimas FEH , SST, FFT e INV foram determinadas a cada quatro dias durante um mês após a poda. Um aumento na atividade da FEH foi observado entre os dias 13 e 20 após a poda, o que coincidiu com o início da brotação dos novos ramos que ocorreu no 13º dia. Este resultado sugere que a despolimerização de frutanos e a brotação são processos concomitantes em V. herbacea, que ocorrem naturalmente durante o ciclo fenológico, mas que também podem ser induzidos em outras fases do ciclo fenológico pela remoção dos ramos aéreos. A FFT parece atuar junto à FEH na diminuição do tamanho das cadeias de frutanos durante a rebrota, enquanto a SST é inibida devido, possivelmente, à interrupção do fornecimento de sacarose aos rizóforos pelos órgãos aéreos. Para a caracterização e purificação parcial da FEH de rizóforos de V. herbacea foram utilizados rizóforos de plantas induzidas à brotação através da remoção dos ramos aéreos. O extrato bruto apresentou, para atividade de FEH, pH ótimo de 4,5, temperatura ótima em 30 ºC e curva sigmoidal de concentração de substrato, sugerindo tratar-se de uma enzima alostérica. Esta enzima também apresentou maior especificidade por ligações do tipo b-2,1 do que sobre ligações b-2,6, e maior afinidade por frutanos de cadeias curtas quando comparadas com frutanos de cadeias longas. Utilizando precipitação com sulfato de amônia, cromatografia de afinidade e cromatografias de troca aniônica e catiônica, quatro frações com atividade de FEH foram purificadas. Destas quatro frações, duas foram submetidas à cromatografia de exclusão molecular, sendo que os pesos moleculares estimados para uma delas foi de 21 kDa e para outra, esta medida situou-se entre 155 e 39 kDa, devido à ampla faixa de exclusão da coluna utilizada. Os pesos moleculares das outras duas frações foram estimados por SDS-PAGE, sendo que as bandas visualizadas corresponderam a 81,3 e 57,5 kDa para uma e 57,5 kDa para a outra fração. / The occurrence of fructans has been reported in native species of a cerrado area of Reserva Biológica e Estação Experimental de Moji Guaçu. Vernonia herbacea, one of these species, presents underground organs named rhizophores which accumulate fructans of the inulin type as reserve carbohydrate. The seasonal growth pattern exhibited by plants of V. herbacea includes sprouting of buds from the rhizophores in spring, followed by a period of flower development and vegetative growth in summer, and dormancy in winter. The fructan content decreases during sprouting and flowering due to mobilization of this carbohydrate during the regeneration of the new shoots. Preliminary studies showed that FEH, the enzyme responsible for the mobilization of fructan, shows high activity only during sprouting. Fructan mobilization was detected by the increase in the amount of reducing sugar released, namely fructose. The aim of this work was to analyze the activities of the enzymes of fructan metabolism, FEH, SST, FFT and invertase and the fructan contents in rhizophores of plants induced to sprouting by defoliation. The enzyme activities were measured every 4 th day for a month after defoliation. Sprouting of new shoots started around the 13 th day, while an increase in FEH activity was detected between 13 and 20 days after defoliation. The results suggest that fructan depolimerization and sprouting are concomitant processes in V. herbacea that occur naturally during the phenological cycle; however, these processes can also be induced by defoliation during other stages of the phenological cycle. FFT seems to act together with FEH by catalyzing the decrease in fructan chain size during shoot regrowth, while SST was inhibited, possibly, due to interruption of sucrose supply to rhizophores from the aerial organs. The characterization and partial purification of FEH were done using rhizophores from plants which were induced to sprouting by defoliation. The optimal pH and temperature for FEH activity were pH4,5 and 30ºC, respectively. The substrate concentration curve exhibited a sigmoid shape, suggesting that FEH of V. herbacea is an alosteric enzyme. Additionally, this enzyme shows more specificity to b-2,1 than to b-2,6 linked fructan and higher affinity for short chain when compared to long chain fructans. Four fractions presenting FEH activity were partially purified by a combination of ammonium sulfate precipitation, affinity chromatography, and anion and cation exchange chromatographies. Two of these were submitted to size exclusion chromatography and the apparent molecular size for one of them was 21 kDa and for the other it was estimated to be between 39 and 155 kDa, due to the wide exclusion limit of the column used. The molecular size of the next two fractions were estimated by SDS- PAGE and the visualized bands corresponded to 81.3 and 57.5 kDa for the first fraction and to 57.5 kDa for the second one.

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