One of the largest problems in current medicine is the shortage of organs for transplant due to technological limitations in the storage of organs for any length of time. A possible solution to this problem would involve cryopreservation. However, current cryopreservatives such as sucrose or DMSO have concerning cytotoxic issues that limit their possible applications. A major cause of cryoinjury is the uncontrolled recrystallization of inter and intra-cellular ice crystals that occurs during the thawing process leading to mechanical damage and dehydration. The Ben lab has thus been interested in the design of compounds that are capable of inhibiting this process but do not possess other undesirable properties found in the native compounds. These synthetic analogues have been shown to increase cellular viability post-thaw. A series of mixed α/β glycopeptides are prepared and analyzed for antifreeze properties. The results of this study imply that it is not the gross conformation of the glycopeptide that is responsible for activity, but rather that intramolecular relationships may be responsible for disrupting the reorganization of ice. A technique was devised for the incorporation of triazoles into the analogues to investigate the importance of the linker and to greatly simplify the synthesis of a library of glycoconjugates. It was found that the IRI activity of glycopeptides is very sensitive to the distance between carbohydrate and peptide backbone. The electron density at the anomeric oxygen is an important parameter with respect to intramolecular networks. A series of substituted galactosides is presented that modify the electronics of the anomeric oxygen. The results demonstrate that decreasing electron density at this position appears to improve IRI activity in a predictable manner. To better understand the remarkable IRI activity of a key analogue, it was systematically truncated. This study led to the serendipitous discovery of a series of very highly IRI active analogues that do not contain a peptide backbone. These compounds represent the first non-glycopeptides that can show very significant IRI activity even at very low concentrations. The final portion of the thesis reports the efforts towards the preparation of a carbasugar analogue of AFGP-8.

Cryopreservation of human red blood cells (RBCs) extends the storage time from 42 days (hypothermic storage limit) to a maximum of 10 years. While this reduces the possibility of RBC shortages in emergency situations, this preservation method is currently limited to individuals with rare blood phenotypes, patients who require autologous blood transfusions, and military applications. Furthermore, cryopreservation is associated with a high degree of cellular damage, which can subsequently reduce the viability of cells post-thaw. The cellular damage incurred upon cryopreservation is primarily attributed to the process of ice recrystallization. To reduce the degree of cellular damage, cryoprotective agents (CPAs) are used. Currently, 10 % dimethyl sulphoxide (DMSO) and 40 % glycerol are used for the cryopreservation of hematopoietic stem cells (HSCs) and human RBCs respectively. Unfortunately, these CPAs do not provide protection against ice recrystallization. The biological antifreezes (BAs) consisting of antifreeze proteins (AFPs) and antifreeze glycoproteins (AFGPs) were identified as the first inhibitors of ice recrystallization. Consequently, the Ben laboratory is interested in synthesizing small molecule carbohydrate-based inhibitors of ice recrystallization that can be used as an alternative to glycerol or DMSO for the cryopreservation of various cell types. Therefore, this thesis focuses on elucidating important structural features of carbohydrate-based derivatives that are responsible for IRI activity. The first part of this study examines the importance of the anomeric oxygen atom of aryl glycosides for IRI activity. Our laboratory previously demonstrated that the O-linked aryl glycosides are effective inhibitors of ice recrystallization. However, the influence of stereoelectronic effects at the C1 position of aryl glycosides on IRI activity has not been investigated. As a result, N- and S-linked aryl glycosides were synthesized in this study and their IRI activities were compared to that of the O-linked aryl glycosides. These results suggest that a stronger exo-anomeric effect exhibited by the C1 nitrogen derivatives reduces the IRI activity of aryl glycosides. The second part of this study focuses on the synthesis of AFGP disaccharide analogs. While the β-(1,3) glycosidic linkage found in native AFGP-8 was previously assessed for its influence on IRI activity, an extensive structure-function analysis of AFGP disaccharide analogs has not yet been performed. As a result, an AFGP disaccharide analog was designed whereby a para-methoxyphenyl (PMP) substituent was incorporated. This was done to assess whether the PMP substituent could enhance the lack of IRI activity exhibited previously with AFGP disaccharide analogs. Although the synthesis of this disaccharide target was not completed, a number of advantageous developments have been made regarding the glycosylation of N-acetyl-D-glycosamine derivatives. In addition, the PMP-GlcNAc intermediate encountered in disaccharide synthesis was assessed for its IRI activity, confirming that the acetamido (NHAc) function is not required for IRI activity.

The development of effective methods to cryopreserve precious cell types has had tremendous impact on regenerative and transfusion medicine. Hematopoietic stem cell (HSC) transplants from cryopreserved umbilical cord blood (UCB) have been used for regenerative medicine therapies to treat conditions including hematological cancers and immodeficiencies. Red blood cell (RBC) cryopreservation in blood banks extends RBC storage time from 42 days (for hypothermic storage) to 10 years and can overcome shortages in blood supplies from the high demand of RBC transfusions. Currently, the most commonly utilized cryoprotectants are 10% dimethyl sulfoxide (DMSO) for UCB and 40% glycerol for RBCs. DMSO is significantly toxic both to cells and patients upon its infusion. Glycerol must be removed to <1% post-thaw using complicated, time consuming and expensive deglycerolization procedures prior to transfusion to prevent intravascular hemolysis. Thus, there is an urgent need for improvements in cryopreservation processes to reduce/eliminate the use of DMSO and glycerol. Ice recrystallization during cryopreservation is a significant contributor to cellular injury and reduced cell viability. Compounds capable of inhibiting this process are thus highly desirable as novel cryoprotectants to mitigate this damage. The first compounds discovered that were ice recrystallization inhibitors were the biological antifreezes (BAs), consisting of antifreeze proteins and glycoproteins (AFPs and AFGPs). As such, BAs have been explored as potential cryoprotectants, however this has been met with limited success. The thermal hysteresis (TH)activity and ice binding capabilities associated with these compounds can facilitate cellular damage, especially at the temperatures associated with cryopreservation. Consequently, compounds that possess “custom-tailored” antifreeze activity, meaning they exhibit the potent ice recrystallization inhibition (IRI) activity without the ability to bind to ice or exhibit TH activity,are highly desirable for potential use in cryopreservation. This thesis focuses on the rational design of potent ice recrystallization inhibitors and on elucidating important key structural motifs that are essential for potent IRI activity. While particular emphasis in on the development of small molecule IRIs, exploration into structural features that influence the IRI of natural and synthetic BAs and BA analogues is also described as these are some of the most potent inhibitors known to date. Furthermore, this thesis also investigates the use of small molecule IRIs for the cryopreservation of various different cell types to ascertain their potential as novel cryoprotectants to improve upon current cryopreservation protocols, in particular those used for the long-term storage of blood and blood products. Through structure-function studies the influence of (glyco)peptide length, glycosylation and solution structure for the IRI activity of synthetic AFGPs and their analogues is described. This thesis also explores the relationship between IRI, TH and cryopreservation ability of natural AFGPs, AFPs and mutants of AFPs. While these results further demonstrated that BAs are ineffective as cryoprotectants, it revealed the potential influence of ice crystal shape and growth progression on cell survival during cryopreservation. One of the most significant results of this thesis is the discovery of alkyl- and phenolicglycosides as the first small molecule ice recrystallization inhibitors. Prior to this discovery, all reported small molecules exhibited only a weak to moderate ability to inhibit ice recrystallization. To understand how these novel small molecules inhibit this process, structure-function studies were conducted on highly IRI active molecules. These results indicated that key structural features, including the configuration of carbons bearing hydroxyl groups and the configuration of the anomeric center bearing the aglycone, are crucial for potent activity. Furthermore, studies on the phenolic-glycosides determined that the presence of specific substituents and their position on the aryl ring could result in potent activity. Moreover, these studies underscored the sensitivity of IRI activity to structural modifications as simply altering a single atom or functional group on this substituent could be detrimental for activity. Finally, various IRI active small molecules were explored for their cryopreservation potential with different cell types including a human liver cell line (HepG2), HSCs obtained from human UCB, and RBCs obtained from human peripheral blood. A number of phenolic-glycosides were found to be effective cryo-additives for RBC freezing with significantly reduced glycerol concentrations (less than 15%). This is highly significant as it could drastically decrease the deglycerolization processing times that are required when RBCs are cryopreserved with 40% glycerol. Furthermore, it demonstrates the potential for IRI active small molecules as novel cryoprotectants that can improve upon current cryopreservation protocols that are limited in terms of the commonly used cryoprotectants, DMSO and glycerol.

Orientador: Telma Teixeira Franco / Tese (doutorado) - Universidade Estadual de Campinas, Faculdade de Engenharia Quimica / Made available in DSpace on 2018-08-07T15:01:29Z (GMT). No. of bitstreams: 1 Tsukamoto_Junko_D.pdf: 1866337 bytes, checksum: 5cac22304ad7fddc116a61d4e57a941f (MD5) Previous issue date: 2006 / Resumo: Reações enzimáticas em meio orgânico têm despertado grande interesse industrial devido às vantagens de obtenção de produtos puros pela maior especificidade das enzimas, pela redução de gastos energéticos e pela possibilidade de desenvolvimento de condições menos agressivas de processo. O uso de enzimas em síntese é uma alternativa viável à síntese orgânica e pode ser compreendida como processo relacionado ao novo conceito de ¿biotecnologia industrial¿ (white biotechnology), devido às preocupações ambientais e da sustentabilidade dos processos químicos. Processos biotecnológicos permitem a exploração racional de materiais renováveis (açúcares) para obtenção de produtos industriais, diminuindo a degradação ambiental. Uma rota enzimática é aqui proposta para obtenção de acrilatos de açúcares, monômero para confecção de polímeros biodegradáveis. O uso de enzimas em síntese orgânica oferece vantagens devido a especificidades na formação de produto, onde os custos de separação, purificação e tratamento de resíduos são minimizados. As lipases são responsáveis por 20% das biotransformações realizadas atualmente. Ésteres derivados de carboidratos e de ácidos graxos vêm sendo produzidos para a indústria alimentícia e farmacêutica, entretanto, mas não se encontram ainda ésteres tais como acrilatos de carboidratos no mercado. Esse projeto visou um estudo da produção de acrilatos de carboidratos pela reação direta via enzimática pela lipase comercial CalB. A esterificação de três monossacarídeos (D-glicose, D-frutose e D-xilose) e dois dissacarídeos (sacarose e maltose) com ácido acrílico pela CalB no solvente terc-butanol foi estudada. O efeito dos três seguintes parâmetros foi avaliado: temperatura, razão molar entre os reagentes (açúcar: ácido acrílico) e quantidade de enzima sobre a conversão a ésteres foi avaliada. A conversão da reação foi acompanhada por cromatografia líquida (CLAE-IR) e por MALDI TOF MS / Abstract: Industrial interest in enzymatic reactions in organic media is due to the possibility of obtain pure compounds with the use of highly specific enzyme, the decrease in energy costs and the possibility of using mild process conditions. The development of new enzymatic processes for chemical synthesis as a viable alternative to the organic synthesis is related to the new concept of "industrial biotechnology" (white biotechnology), with higher concerns with the environment and the sustainability of chemical processes. Biotechnological processes that make a more rational use of renewed resources such as sugars to produce chemicals decrease, in most cases, the environmental impact. In this work an enzymatic route is proposed for the production of sugar acrylates, a biodegradable monomer. The use of enzymes in organic media has the advantage of high specificity to the desired product, lower separation and purification costs, and minimization of residue formation. Currently, lipases are responsible for 20% of all biotransformations. Esters derived from fatty acid and carbohydrates are produced for the food and pharmaceutical industries. However, acrylate esters of carbohydrate are still not found in the market. This aim of this work was to study the production of acrylates of carbohydrates by the direct enzymatic reaction of sugars with acrylic acid on the commercial lipase CalB. The esterification of three monosaccharide (D-glucose, D-fructose and xylose) and two disaccharides (sucrose and maltose) with acrylic acid on CalB in tert-butanol was studied. The effects of temperature, molar ratio of the reactants (sugar to acrylic acid) and amount of enzyme on the conversion to esters were studied. The esterification reaction was monitored by HPLC and MALDI TOF MS / Doutorado / Desenvolvimento de Processos Químicos / Doutor em Engenharia Química

Orientadores: Glaucia Maria Pastore, Luciana Gomes Malta / Dissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Engenharia de Alimentos / Made available in DSpace on 2018-08-27T06:04:46Z (GMT). No. of bitstreams: 1 Pavan_Veronica_M.pdf: 1806354 bytes, checksum: 1b5d278303881581f3b4c456b216d4a5 (MD5) Previous issue date: 2015 / Resumo: Compostos com atividade antioxidante provenientes de frutas que estão envolvidos na prevenção de várias doenças degenerativas foram investigados em diversos estudos. Para melhorar o conhecimento do processo de absorção desses compostos com atividade antioxidante (AA), este estudo avaliou a AA de extratos de Artocarpus heterophillus (jaca) e Annona marcgravii (araticum) brutos, digerido gástrico, digerido intestinal e fermentação colônica, in vitro. Os compostos fenólicos foram avaliados a partir da quantificação de fenóis totais (FT), flavonoides totais (FLT), seguida da AA, medida por métodos como Trolox equivalent antioxidant capacity (TEAC) e Oxygen radical absorbance capacity (ORAC). Um importante fator que tem chamado atenção desta classe de compostos são os fenólicos que estão ligados a diferentes moléculas, como carboidratos, aminoácidos, dentre outros, que pelo seu peso molecular, tipo de ligação podem ser absorvidos ou mesmo metabolizados em diferentes regiões no sistema gastrintestinal. No extrato de araticum foi observada uma redução em FT após a digestão gástrica e subsequente aumento após a digestão intestinal, seguida de redução após a fermentação. Redução e aumento em FLT foram observados no extrato de jaca. A AA apresentou diferentes resultados para os métodos utilizados. Observou-se um aumento em TEAC para o extrato de jaca após o processo de digestão intestinal, diferente do extrato de araticum que apresentou aumento após a digestão gástrica e redução após a fermentação. Na avaliação pelo método ORAC observou-se um aumento do potencial no extrato de jaca após sofrer o processo de fermentação colônica. O extrato de araticum apresentou diferente comportamento quando comparado ao extrato de jaca, mostrando queda sucessiva na AA quando digerido e também após ser fermentado. Os carboidratos apresentaram um perfil interessante, no qual desvia do padrão estabelecido pela literatura mostrando uma redução após a digestão gástrica / Abstract: Compounds with antioxidant activity from fruits that are involved in the prevention of several degenerative diseases were investigated in several studies. To improve the knowledge of the process of absorption of these compounds with antioxidant activity (AA), this study evaluated the crude extracts of the Artocarpus heterophillus (jackfruit) and Annona marcgravii (araticum), gastric and intestinal digested and colonic fermentation in vitro. Phenolic compounds were evaluated from the quantification of total phenols (FT), total flavonoids (FLT), followed by AA, measured by method such as Trolox equivalent antioxidant capacity (TEAC) and Oxygen radical absorbance capacity (ORAC). An important factor which has been highlighted to this class of phenolic compounds are the phenolics which are attached to various molecules such as carbohydrates, amino acids, among others, that at molecular weight, type of linkage can be absorbed or metabolized in different regions of the gastrointestinal system. In the araticum extract was observed a decrease in FT after gastric digestion and afterwards an increase after intestinal digestion, followed by reduction after the fermentation. Reduction and increase in FLT were observed in jackfruit extract. The AA showed different results for the method used. There was an increase in TEAC for jackfruit extract after the process of intestinal digestion, unlike araticum extract which increased after gastric digestion and reduction after the fermentation. In assessing the ORAC method there was an increased in the potential of the jackfruit extract after suffering the colonic fermentation process. The araticum extract showed different behavior compared to jackfruit extract, showing successive reduction in AA when it was digested and also after being fermented. Carbohydrates showed an interesting profile, which deviates from the standard established in the literature showing a reduction after gastric digestion / Mestrado / Ciência de Alimentos / Mestra em Ciência de Alimentos

O cromo é o sétimo metal mais abundante na crosta terrestre e ocorre naturalmente nos diversos compartimentos ambientais, porém o intenso uso deste metal em atividades industriais tem aumentado a liberação deste contaminante, ocasionando alterações na estrutura e funcionamento dos ecossistemas terrestres e aquáticos. Considerando as diferentes formas de exposição ao metal pelos organismos, procurou-se avaliar os efeitos do cromo nas populações fitoplanctônica e zooplanctônica e nas interações tróficas entre ambas, com base em aspectos estruturais e fisiológicos. Para tanto, foram realizados ensaios ecotoxicológicos com duas espécies de algas (Pseudokirchneriella subcapitata e Chlorella vulgaris), avaliando efeitos na densidade celular, concentração de pigmentos e composição bioquímica, além de ensaios com organismos zooplanctônicos (Cladocera Ceriodaphnia dubia e Ceriodaphnia silvestrii) expostos a diferentes vias de contaminação, como água contaminada, alimento contaminado e a combinação de ambas as situações (água e alimento contaminados), avaliando efeitos na sobrevivência, fecundidade, taxa de alimentação e composição bioquímica. Os resultados obtidos demonstraram que o cromo, nas condições testadas, afetou o crescimento, a concentração de pigmentos e a composição bioquímica das algas P. subcapitata e C. vulgaris. Nos ensaios realizados com C. dubia e C. silvestrii, os diferentes tratamentos com cromo não alteraram a sobrevivência destas espécies, porém, nos tratamentos com meio aquoso contaminado com cromo e exposição combinada (meio aquoso e alimento contaminados com cromo) houve redução na fecundidade e diminuição da taxa alimentar destes organismos. Redução no crescimento populacional e aumento de proteínas, carboidratos e lipídeos totais de C. dubia e C. silvestrii também foram relacionados à presença de cromo. Conclui-se, portanto, que o cromo, nas condições testadas, foi tóxico para as espécies fito e zooplanctônicas, podendo causar alterações nas cadeias tróficas dos ecossistemas aquáticos. / Chromium is the seventh most abundant metal in the earth\'s crust and occurs naturally in various environmental compartments, but the heavy use of this metal in industrial activities has increased the release of this contaminant, causing changes in the structure and functioning of terrestrial and aquatic ecosystems. Considering the different forms of exposure to metal by organisms, sought to evaluate the effects of chromium on phytoplankton and zooplankton populations and trophic interactions between them, based on structural and physiological aspects. For this, ecotoxicological tests were conducted with two species of algae (Pseudokirchneriella subcapitata and Chlorella vulgaris), assessing effects on cellular density, pigment concentration and biochemical composition, and tests with zooplankton (Cladocera Ceriodaphnia dubia and Ceriodaphnia silvestrii exposed to different ways of contamination, such as contaminated water, contaminated food and a combination of both situations (contaminated water and food), evaluating effects on survival, fertility, feed rate and biochemical composition. The results showed that chromium under the conditions tested, affected the growth, pigment concentration and biochemical composition of the algae P. subcapitata and C. vulgaris. For tests with C. dubia and C. silvestrii, the different treatments with chromium did not affect survival of these species, however, in treated aqueous medium contaminated with chromium and combined exposure (aqueous and food contaminated with chromium) there was a reduction in fecundity and decreased feed rate of these organisms. Reduction in population growth and increased protein, carbohydrate and lipid of C. dubia and C. silvestrii were also related to the presence of chromium. Therefore, it is concluded that chromium, in the conditions tested, was toxic to phytoplankton and zooplankton species, may cause changes in trophic chains of aquatic ecosystems.

O ambiente epifítico é considerado um dos habitats mais desafiadores para as plantas, pois a disponibilidade hídrica e nutricional pode ser bastante escassa. Além disso, as plantas que colonizam este ambiente estão mais expostas aos fatores ambientais, dentre eles a luminosidade se mostra bastante importante, uma vez que esta atua tanto como fonte de energia na fotossíntese quanto como sinal ambiental em repostas fotomorgênicas. Sabe-se, por exemplo, que a luz exerce forte influência sobre a morfogênese radicular de plantas em geral, porém impacta ainda mais o desenvolvimento de orquídeas epífitas, dada a frequente exposição de suas raízes aéreas à incidência luminosa. Tendo em vista que a auxina ácido indolil-3-acético (AIA), o ácido abscísico (ABA) e o etileno são moduladores cruciais no controle da arquitetura radicular na maioria das plantas, sendo também mediadores chave em várias respostas fotomorfogênicas, este estudo propôs-se a investigar o possível envolvimento destes hormônios durante diferentes respostas morfo-fisiológicas desencadeadas pela exposição à luz do sistema radicular de plantas de C. fimbriatum. A ausência de incidência luminosa sobre às raízes resultou em maiores taxas de crescimento e volume radicular, porém, com menor acúmulo de biomassa em relação às raízes expostas à luz. O incremento na biomassa em raízes expostas à luz esteve correlacionado ao espessamento da parede celular na região cortical, o qual ocorreu em resposta especificamente à luz azul. Em termos gerais, a exposição das raízes à luz induziu o aumento nos níveis de AIA e ABA, enquanto que os teores de ACC foram superiores em raízes protegidas da incidência luminosa. Estes resultados sugerem que a luz pode modular o desenvolvimento radicular de C. fimbriatum através de um fino controle hormonal que depende de ajustes coordenados dos níveis de AIA ABA e ACC. Também foi investigado o potencial envolvimento das auxinas e do ABA durante a remobilização de carboidratos entre pseudobulbos e folhas de plantas que tiveram seus sistemas radiculares expostos à (ou protegidos da) luz. Os resultados revelaram que a manutenção das raízes sob condições de escuro levou ao aumento dos teores de AIA e de todas as fontes de carbono estudadas (especialmente de glicose e frutose) nos pseudobulbos, enquanto que as raízes cobertas apresentaram apenas um leve aumento no conteúdo de AIA. O tratamento concomitante das raízes com a condição de escuro e a aplicação de um inibidor do transporte polar de auxina causou uma diminuição abrupta nos teores de AIA em todos os órgãos analisados e a elevação do conteúdo de ABA no sistema radicular. De maneira interessante, essa última condição experimental induziu um conspícuo acúmulo de carboidratos nos pseudobulbos, principalmente de sacarose. Assim, os dados deste trabalho reforçam a importante participação do AIA e ABA como possíveis mediadores da sinalização desencadeada pela luz incidente no sistema radicular de C. fimbriatum, cujas respostas induzidas regulam não somente a morfogênese de tecidos radiculares, mas também influenciam na regulação da partição de carbono no sistema caulinar por meio de um provável mecanismo de sinalização à longa distância / The epiphytic environment is considered one of the most challenging for plants, due to frequent scarcity of water and nutrients. Furthermore, the plants that colonized this biotope are usually more exposed to the environmental cues. Light is considered one of the most important signals controlling plant development because it can act as both an energy source for photosynthesis and an environmental signal for photomorphogenic responses. Besides, light can influence the root morphology of most plants, with even stronger impacts expected in aerial roots of epiphytic orchids due their frequent exposition to direct light. Since indole-3-acetic acid (IAA), abscisic acid (ABA) and ethylene are crucial hormonal signals modulating the root architecture in most plant species, and key mediators during numerous photomorphogenic responses, this study investigated the potential involvement of these hormones in different morpho-physiological responses regulated by either the darkness treatment or the light exposure of Catasetum fimbriatum root system. The absence of light incidence on the roots resulted in higher root volume and growth rate, but lower dry mass accumulation than the light-exposed ones. The higher accumulation of biomass in the light-exposed roots was closed correlated with a more intense cell wall thickening in the root cortex, which appeared to be specifically induced by the blue light. In general, root exposure to light induced increasing levels of ABA and AIA, while the ACC content was higher in roots protected from light. This suggests that light might modulate C. fimbriatum root development through a fine-tuned hormonal mediation, which depends on coordinated adjustments of IAA, ABA and ACC levels. This study also investigated the potential involvement of auxin and ABA during the (re)mobilization of carbohydrates in pseudobulbs and leaves of plants that had their root systems either exposed to (or protected from) light. The results revealed that covering the roots increased in pseudobulbs the levels of AIA and all carbon sources studied (specially glucose and fructose), while the covered roots showed slightly higher levels of AIA. The concomitant treatment with NPA and root covering caused a sharp decrease of AIA levels in all organs and an ABA increase in the root system. Interestingly, this last condition induced a conspicuous carbohydrate accumulation in pseudobulbs, with sucrose as the predominant form. The data obtained in this study reinforce the remarkable participation of IAA and ABA as possible mediators of the signaling cascades triggered by the light incidence on C. fimbriatum root system, which was capable of inducing photomorphogenic responses not only in root tissues, but was also able to influence the carbon portioning in the shoot system by a potential long-distance signaling mechanism

Plantas típicas de regiões alagáveis tendem a apresentar diversas estratégias morfológicas e fisiológicas a fim de otimizar as chances de sobrevivência nestes ambientes. Um dos principais mecanismos, porém menos estudados, é a plasticidade encontrada no metabolismo de carboidratos que está diretamente ligado a alta demanda energética inerente a estes locais. Senna reticulata (Leguminosae) é uma espécie pioneira típica de planícies sujeitas ao regime anual de inundação da Amazônia Central, considerada uma das mais eficientes colonizadoras destes ambientes, além de extremamente tolerante ao alagamento. Neste trabalho, pela primeira vez foram caracterizados os padrões de reserva e de mobilização de açúcares desta espécie no período que corresponde ao seu estabelecimento no início da sua primeira fase terrestre. Adicionalmente o efeito da elevada concentração de gás carbônico foi verificada cultivando-se as plantas em câmaras de topo aberto (OTCs) em concentrações de CO2 ambiente (~380 &#181;mol) e elevado (~760 &#181;mol). Foram realizadas coletas destrutivas para análises de conteúdo de carboidratos não estruturais ao longo de 90 dias e ao longo de 24 horas. Foi verificado que S. reticulata possui altos níveis de amido foliar indicando que essa reserva pode estar diretamente relacionada a estratégia de constante produções de novas folhas que esta espécie apresenta. As razões sacarose:monossacarídeos se apresentaram altas em todos os órgãos analisados especialmente na raiz indicando que esta espécie pode apresentar estratégias metabólicas relacionadas também a estação seca pois além da sacarose poder ser utilizada como principal substrato na formação de raízes adventícias em condições de alagamento, também pode ser útil na regulação osmótica da raiz em locais com solos que apresentam baixa disponibilidade hídrica. A elevada concentração de gás carbônico promoveu um aumento significativo das reservas de amido na folha e no caule além de diminuir a taxa de degradação de amido transitório durante o período noturno. As concentrações de açúcares solúveis (glicose, frutose e sacarose) apresentaram tendência de diminuição sob condições de atmosfera enriquecida provavelmente devido à maior utilização dos mesmos para o crescimento da raiz. Este conjunto de respostas indica que essa espécie responde de forma positiva a elevada concentração de gás carbônico utilizando-se de possíveis mecanismos que diminuem a sinalização mediada por açúcares como, por exemplo, a habilidade de aumentar a capacidade de seus drenos já existentes, promovendo a mobilização do excesso de carboidratos para reservas e crescimento. / Plants of floodplains tend to show many morphological and physiological strategies to get higher survival rates in this places. One of them but with received less attention is the plasticity showed by carbohydrate metabolism that is directly linked to high energy demand at this sites. Senna reticulata (Leguminosae) is a pioneer tree which is common in floodplains sites submitted to an annual pulse of flooding at Central Amazonian and one of the most efficient colonizers at this sites besides extremely flooding tolerant. In this work by the first time was made the characterization about reserves and mobilizations patterns of sugars on the time range that corresponding to the establishment at the initial first terrestrial phase. Additionally, the effect of high CO2 was checked by growing plants in Open-Top-Chambers (OTC´s) with ambient (~380 &#181;mol) and elevated (~760 &#181;mol) CO2 concentrations. It was made systematic harvests along periods of 24 hours and 90 days to determinate the carbohydrate non-structural content. It was founded that S. reticulata had higher levels of starch in leaves showing evidences that this reserve is directly linked with constantly production of new leaves strategy. The ratio sucrose:monosaccharides was high in all organs but specially in roots showing evidences that this specie has metabolic strategies also related with drought periods because besides sucrose could be used as the aim substrate to adventitious root formation at flooding conditions and can be useful too on the osmotic regulation of roots in sites with low moisture content soil. The high levels of carbon dioxide promote significative increase of starch in leaves and stem besides decrease the degradation rate of transitory starch in leaves in the dark period. The levels of soluble sugars (glucose, frutose and sucrose) showed tendency of decrease under high CO2 conditions probably due to higher use in root growth. This set of answers evidence that this tree has positively response to high CO2 useful by possible mechanisms that decrease the signaling through sugars like for example the big abiliity of increase capacity of preexistents sinks to promoting the surplus carbohydrates mobilization to reserves and growth.

<p>Moderating glycemic response to foods is important for the potential to control or prevent hyperglycemia-related diseases, such as diabetes and cardiovascular disease. The importance of slowly digestible carbohydrates (SDC) lies in their health effects: moderated blood glucose response, and a potential for increased satiety and reduced intake, and weight management. The research presented is on structural properties of novel, mostly soluble, a-glucans (glucose-containing oligomers and polymers with different linkage types and combinations) that are required for slow yet full digestion, and how they behave in food systems. Up to this point, little has been known regarding what structural properties of glucose-containing carbohydrates result in slow digestion, although starch structure has been well investigated and it is known that raw starch has a slowly digestible property. In addition to the structure-function aspect of the thesis work, this research contributes information about how a-glucan SDCs can be incorporated into food products that undergo heat treatment in the presence of moisture. The a-glucans maintain their SDC property while raw starch is gelatinized and becomes rapidly digestible. The rate of hydrolysis of a large number of novel a-glucans was studied using a simulated upper gastrointestinal in vitro digestion utilizing porcine pancreatic α-amylase and α-glucosidases from the rat intestine, and a subset was then evaluated in a crossover design clinical trial with blood glucose monitoring. Linkage and molecular weight analysis using gas chromatography of partially methylated alditol acetates and multi-angle light scattering and refractive index (MALS-RI) detection at time points during in vitro digestion were used to elucidate the relative rate of digestion of different linkage types in new and known a-glucan carbohydrates. Rheological, turbidity, and SLS and DLS analyses were used to examine ingredient interaction between novel, enzymatically-modified α-glucans with slow digesting properties found most promising for inclusion in food products. A model nutritional beverage system was utilized containing proteins and salts. It was found that solvent and ion concentration of solutions were important for dictating aggregation formation with highly branched alternans and oligosaccharides in solution alone, or in the presence of soluble protein aggregates. Further, salts in solution proved to influence rheological and turbidity measures of all four α-glucans examined in the model system, indicating they may affect aggregation and structural conformation of such large carbohydrates. However, only tapioca maltodextrins showed in vitro rate of digestion affected by aggregation.</p><p></p> <p>These results show the chemical and molecular properties of modified carbohydrates that contribute to slow digestion rate, which is informative to develop improved or new SDCs, as well as how these novel SDCs interact with other ingredients within a model beverage system, informing applications for the food industry.</p>

This master thesis deals with the investigation of selected chemical and sensory characteristics of ciders, which were made from four selected apple varieties. The theoretical part describes the chemical composition of apples, cider, the technology of production and production in the world. The next section describes the analytical methods used to determine chemical characteristics not only in ciders but also in other alcoholic beverages. The experimental part focuses on the determination of chemical characteristics such as the content of organic acids, carbohydrates, total phenolic content, alcohol antioxidant activity and elemental composition. The purpose of the work was to find out how different varieties differ in chemical composition. The produced ciders were subjected to sensory analysis to evaluate the selected cider sensory parameters, which were in the natural state and with the addition of carbon dioxide. The results show that ciders of different varieties differ in chemical composition. Malic acid was the most cider's abundant of the organic acids in the range of 2759 – 4411 mg·l-1. Another acid represented was lactic acid in the range of 166,2 – 288,1 mg·l-1. The glucose content of each cider was determined between 845,3 – 1159,8 mg·l-1and fructose 32,6 – 406,7 mg·l-1. Potassium 944,29 – 1073,98 mg·l-1 was the most represented of all elements. The total phenolic content was determined to be in the range of 164,2 – 214,1 mg of GAE·l-1and the antioxidant activity of 0,192 – 0,318 mg·l-1. These differences in chemical composition were also reflected in sensory evaluation. According to the overall assessment, the best cider was made from the Granny Smith variety, which contained the most organic acids and carbohydrates. It has been found that by adding carbon dioxide, sensory parameters such as colour, taste and smell are altered.