The present study was initiated to determine the effect sorbitol has upon the enzymatic functions of thiamin; specifically, what effect sorbitol bas upon the alpha-ketoglutarate and pyruvate dehydrogenases activities of rat liver mitochondria. In addition, the effect of varying the dietary sorbitol concentration on the development of an athiaminosis was studied, particularly the effect on growth rate.

No / Two force fields, the GROMOS53A5/53A6 (united atom) and the AMBER95 (all atom) parameter sets, coupled with partial atomic charges derived from quantum mechanical calculations were evaluated for their ability to reproduce the known crystalline forms of the polyols mannitol and sorbitol. The force fields were evaluated using molecular dynamics simulations at 10 K (which is akin to potential energy minimization) with the simulation cell lengths and angles free to evolve. Both force fields performed relatively poorly, not being able to simultaneously reproduce all of the crystal structures within a 5% deviation level. The parameter sets were then systematically optimized using sensitivity analysis, and a revised AMBER95 set was found to reproduce the crystal structures with less than 5% deviation from experiment. The stability of the various crystalline forms for each of the parameter sets (original and revised) was then assessed in extended MD simulations at 298 K and 1 bar covering 1 ns simulation time. The AMBER95 parameter sets (original and revised) were found to be effective in reproducing the crystal structures in these more stringent tests. Remarkably, the performance of the original AMBER95 parameter set was found to be slightly better than that of the revised set in these simulations at 298 K. The results of this study suggest that, whenever feasible, one should include molecular simulations at elevated temperatures when optimizing parameters.

Bacteria of the genus Gluconobacter have very active, membrane-bound, NAD(P)-independent, polyol dehydrogenases which stoichiometrically produce the singlestep oxidation product of polyols provided in the growth medium. These bacteria have a high respiratory quotient which is believed to result from oxidations by these dehydrogenases. These organisms grow and survive at pH values as low as 2.5 leading to speculation that their membrane-bound dehydrogenase activity provides the rapid electron flow necessary to purge cells of toxic levels of hydrogen ions. These dehydrogenases are also believed to be used for energy metabolism, and there is no clear understanding of their function in the cell metabolism. Oxidation of sorbitol in Gluconobacter oxydans ATTC 621 was studied to determine if the oxidations by the membrane bound sorbitol dehydrogenase (mSDH) were required for growth, and whether they functioned to protect the cells in low pH environments. G. oxydans required a high concentration of sorbitol in the medium, and a reduction in the concentration to 0.1% decreased the rate and extent of growth. Using mutants with decreased levels of mSDH, we found that growth rates decreased as a result of this mutation, indicating that mSDH activity was needed for growth. No changes in the specific activity of mSDH in strain ATCC 621 occurred when the cells were grown at pH 7.0, 6.0, and 4.5. However, cytochrome levels were doubled in cells grown at pH 4.5 compared to pH 6.0 and 7.0. The increased cytochrome levels did not increase the oxygen uptake of the pH 4.5 grown cells on sorbitol. Cells grown at all pH values respired more rapidly when tested at pH 4.5, and respiration decreased as the pH increased. The higher activity at lower pH values may result from increased efficiency of mSDH, which has an in vitro pH optimum of 5.2. Magnesium and calcium increased the respiration of pH 6.0 grown cells but not pH 4.5 grown cells. Less cell mass per mg of sorbitol oxidized was obtained when cells were grown at pH 4.5 compared to pH 6.0 and 7.0. However, no differences were detected in the specific activity of any of the sorbitol oxidizing enzymes. The activity of mSDH in G. oxydans is necessary for the growth of this bacterium. The mSDH specific activity is not regulated by the growth pH, but increased levels of cytochromes and decreased cell yields indicate a change in the cell's oxidative system resulting from lowered growth pH values. / Ph. D.

La prééclampsie (PE) est une pathologie obstétricale complexe associée à un défaut de placentation. Selon la littérature, le placenta anormalement développé libèrerait des facteurs qui induiraient une dysfonction endothéliale maternelle. Nous avons émis comme hypothèse qu’une accumulation de sorbitol, un sucre hyperosmotique produit par la voie des polyols, pourrait induire la libération par le placenta de sFlt-1, un facteur anti-angiogénique antagoniste du VEGF en PE. Nous avons comparé les niveaux d’expression d’ARNm et de protéines de AKR1B1 et SORD, les deux enzymes impliquées dans la voie des polyols, dans les placentas de mères ayant eu une grossesse normotensive (groupe témoin) ou prééclamptique par RT-PCR quantitatif et par immunobuvardage respectivement. Nous les avons ensuite localisées par immunohistochimie. Nos résultats suggèrent que la voie des polyols serait altérée au niveau de la membrane amniochorionique des placentas issus de grossesses prééclamptiques, ce qui pourrait favoriser une accumulation de sorbitol à l’interface fœto-maternelle. / Preeclampsia (PE) is a complex obstetrical pathology associated to a defective placentation. The abnormally developed placenta is believed to release factors causing a maternal endothelial dysfunction. We hypothesized that an accumulation of sorbitol, a hyperosmotic sugar produced through the polyol pathway, could induce the release by the placenta of sFlt-1, an antagonist of the angiogenic factor VEGF in PE. We compared mRNA and protein expression levels of AKR1B1 and SORD, the two enzymes of the polyol pathway, in placentas from normotensive (control) and PE pregnancies by quantitative RT-PCR and immunoblotting respectively. Then, we localized the two enzymes by immunohistochemistry. Our results suggest that polyol pathway is altered in amniochorionic membranes from PE pregnancies, and that this phenomenon would promote sorbitol accumulation at the foeto-maternal interface.

O interesse na aplicação de embalagens ativas para conservação de alimentos tem aumentado, bem como uma maior demanda por substâncias antimicrobianas naturais, devido à maior consciência dos consumidores quanto aos potenciais riscos à saúde ocasionados pelo consumo de compostos sintéticos. Entre os aditivos naturais, a própolis, por suas propriedades antibacteriana, antioxidante e antifúngica, tem despertado o interesse dos pesquisadores. Portanto, o objetivo deste trabalho foi investigar o efeito da adição do extrato etanólico de própolis (EEP) em filmes à base de gelatina plastificados com citrato de acetiltributila (CA) ou sorbitol (S), nas propriedades funcionais (propriedades mecânicas, solubilidade, permeabilidade ao vapor de água, parâmetros de cor e opacidade) e a atividade antimicrobiana dos filmes contra Staphylococcus aureus. Para a produção do EEP, 30g de resina de própolis (tipo 12) foram misturadas com 100mL de álcool etílico 80%, e a solução foi mantida sob agitação mecânica (500rpm, 30minutos) a 50ºC, e filtrada após 24horas sob refrigeração. Os filmes foram produzidos por casting com 2g de gelatina/100g de solução filmogênica, 30g de CA ou S/100g de gelatina, 35g de lecitina/100g de plastificante e o EEP nas concentrações de 0, 5, 40 ou 200g/100g de gelatina, e avaliados quanto às suas propriedades mecânicas (tração e perfuração), solubilidade em água (Sol), cor e opacidade, permeabilidade ao vapor de água (PVA), espectroscopia de infravermelho com transformada de Fourier (FTIR) e microscopia eletrônica de varredura, após acondicionamento (5 dias, UR = 58%, T = 25ºC). A atividade antimicrobiana dos filmes contra Staphylococcus aureus foi analisada pelo método da difusão em ágar. A incorporação de EEP causou variação na tensão (T) dos filmes com CA (58,7 a 66,4MPa), mas afetou mais intensamente os filmes com S (31,7 a 50,4MPa). Quanto à elongação (%), os filmes (CA e S) também apresentaram variações. Na perfuração, verificou-se que a adição de EEP não afetou significativamente a força máxima para ambos os plastificantes. Para a solubilidade em água, o filme com CA apresentou aumento significativo (Sol = 18,6%) para 200% de EEP, contudo, os filmes com S praticamente não variaram. Quanto à cor, para ambos os plastificantes estudados (CA e S) o aumento da concentração de EEP promoveu alterações significativas de L*, a*, b*, ΔE* e opacidade. As análises de PVA dos filmes (CA e S) mostraram que o aumento na concentração de EEP aumentou a propriedade de barreira dos filmes. Segundo a análise de FTIR, os filmes aditivados não apresentaram mudanças estruturais em relação ao controle, o que sugere que o EEP encontra-se disperso na matriz. As micrografias mostraram que a adição de EEP causou alterações, principalmente, nos filmes plastificados com S. A atividade contra S. aureus foi observada para os filmes (CA e S) com 40 e 200% de EEP, com aumento significativo do diâmetro de inibição em função do aumento da concentração de EEP. Os filmes mantiveram sua atividade antimicrobiana durante 177 dias de armazenamento. Estes resultados demonstraram a capacidade antimicrobiana dos filmes de gelatina aditivados com própolis, bem como sua estabilidade em função do tempo. / The interest in the use of active packaging for food conservation has increased, along with a higher demand for natural antimicrobial substances, due to higher consumer awareness of the potential health risks caused by synthetic compounds consumption. Among natural additives, propolis, has awaken the interest of the researchers for its antibacterial, antioxidant and antifungal properties. Thus, the aim of this work was to investigate the effect of the addition of an ethanolic propolis extract (EPE) to gelatin-based films plasticized with acetyltributyl citrate (AC) or sorbitol (S), on the functional properties (mechanical properties, solubility, water vapor permeability, color parameters and opacity) and the antimicrobial activity of the films against Staphylococcus aureus. For the EPE production, 30g for the propolis resin (type 12) were mixed with 100mL ethyl alcohol (80%), and the solution was maintained under mechanical stirring (500rpm, 30minutes) at 50°C, and filtered after 24hours under refrigeration. The films were produced by casting, using 2g of gelatin/100g of film forming solution, 30g of AC or S/100 g of gelatin, 35g of lecithin/100g of plasticizer and EPE in the concentrations of 0, 5, 40 or 200g/100g of gelatin, and evaluated for their mechanical properties (traction and puncture tests), water solubility (Sol), color and opacity, water vapor permeability (WVP), Fourier transform infrared spectroscopy (FTIR) and scanning electron microscopy, after storage (5 days, relative humidity = 58%, temperature = 25°C). The antimicrobial activity of the films against Staphylococcus aureus was analyzed by agar diffusion method. The EPE addition caused variation of the tensile strength (T) of the films with AC (58.7 a 66.4MPa), but affected more intensely the films with S (31.7 a 50.4MPa). The films (AC and S) also presented variations for the elongation (%). For the puncture force, the addition did not affect significantly the maximum force for any of the plasticizers. For the water solubility, the films with AC presented significant increase (Sol = 18,6%) for 200% of EPE, however the films with S did not vary. For both plasticizers studied (AC and S), the increase of EPE concentration promoted significant changes of the color parameters, L*, a*, b*, ΔE* and opacity. The WVP analysis of the films (AC and S) showed that the increase of the EPE concentration increased the barrier property of the films. According to FTIR analysis, the EPE addition in the films did not present structural changes compared to the control, suggesting that the EPE was disperse in the protein matrix. The micrographs showed that the EPE addition caused changes mainly in films plasticized with S. The activity against S. aureus was observed for the films containing 40 and 200% of EPE, with significant increase of inhibition diameter with increase of EPE concentration. The films kept their antimicrobial activities during 177 days of storage. These results demonstrated the antimicrobial capacity of gelatin-based films added of propolis, and their stability in time.

Apesar dos conhecimentos já existentes sobre o xilitol, seu efeito no desenvolvimento da cárie dentária ainda requer maiores esclarecimentos. Assim, este estudo in situ randomizado, duplo-cego e cruzado teve como objetivo determinar o efeito de xilitol em pastilhas na composição do biofilme dental e nos processos de desmineralização e remineralização do esmalte, comparado a um controle com pastilhas contendo sorbitol. Em duas fases de 14 dias, 11 voluntários utilizaram dispositivos palatinos com seis blocos de esmalte humano (três hígidos para avaliação da desmineralização + três previamente desmineralizados para avaliação da remineralização) com dureza de superfície conhecida. Sacarose 20% foi gotejada oito vezes ao dia apenas sobre os blocos hígidos. Cinco minutos após cada gotejamento, os dispositivos eram recolocados na boca e os voluntários chupavam pastilha com xilitol 88,3% ou sorbitol 84,5%. A utilização das pastilhas iniciou-se uma semana antes de cada fase experimental. Foi usado dentifrício sem flúor. Ao final de cada fase, o biofilme formado sobre os blocos foi coletado e dividido para análises microbiológica e bioquímica, e os fragmentos de esmalte foram analisados quanto a variação de dureza de superfície antes e após o experimento. A porcentagem de perda de dureza de superfície dos blocos de esmalte do grupo do xilitol apresentou uma tendência a ser menor do que aquela observada no grupo do sorbitol, embora a diferença não tenha sido significante (p= 0,066; ANOVA). Também não houve diferença estatisticamente significante entre os grupos em relação à porcentagem de ganho de dureza de superfície e à maioria dos parâmetros bioquímicos do biofilme. As médias das concentrações de fósforo inorgânico e de polissacarídeo intracelular no biofilme no experimento de desmineralização foram significantemente menores (p< 0,001 e p= 0,007) no grupo do sorbitol. As porcentagens de estreptococos do grupo mutans (SM) em relação ao total de estreptococos (S) (p= 0,037) no experimento de desmineralização, assim como as contagens de SM (p= 0,035) e lactobacilos (p= 0,048), e as porcentagens de SM em relação ao total de microrganismos (p= 0,035) e em relação a S (p= 0,017) no experimento de remineralização foram significantemente menores no biofilme do grupo do xilitol. Os demais parâmetros microbiológicos não foram influenciados de maneira significante pelos tratamentos. O uso de pastilhas com xilitol por curtos períodos de tempo não apresentou vantagem nem em diminuir a desmineralização nem em favorecer a remineralização do esmalte em comparação ao uso de pastilhas com sorbitol, mas alterou a ecologia bacteriana do biofilme, reduzindo as contagens e as porcentagens de importantes grupos de microrganismos cariogênicos. / In spite of the existing knowledge about xylitol, its effect on the development of dental caries is still a subject that requires further clarifications. Thus, this randomized, double-blind, crossover in situ study aimed at assessing the effect of xylitol mints in the dental biofilm composition and in the processes of enamel demineralization and remineralization, in comparison to a control group using sorbitol mints. During the two phases of 14 days, 11 volunteers wore palatal appliances with six human enamel blocks (three intact ones for evaluation of demineralization and three demineralized ones for the evaluation of remineralization) of known superficial hardness. A solution of 20% sucrose was dripped only on the intact enamel blocks eight times per day. Five minutes after each dripping, the appliances were placed back inside the mouth and the volunteers took either an 88.3% xylitol mint or an 84.5% sorbitol mint. Both types of mints started being used by the volunteers one week before each experimental phase. Non-fluoridated toothpaste was used. At the end of each phase, the biofilm formed over the blocks was collected and divided for microbiological and biochemical analysis and the enamel fragments were evaluated in relation to surface hardness variation before and after the experiment. The percentage of surface hardness loss of the enamel blocks in the xylitol group tended to be lower than in those of the sorbitol group although this difference was not significant (p= 0.066; ANOVA). Moreover, there were no statistically significant differences between the study groups regarding the percentage of surface hardness gain or regarding the biochemical parameters of the biofilm. The average concentrations of inorganic phosphorus and of intracellular polysaccharides in the biofilm for the demineralization experiment were significantly lower (p< 0.001 and p= 0.007) in the sorbitol group. The following results were significantly lower in the xylitol group biofilm: percentages of mutans streptococci (SM) in relation to the total number of streptococci (S) (p= 0.037) in the demineralization experiment; SM (p= 0.035) and lactobacilli (p= 0.048) count; and the percentages of SM in relation to the total number of microorganisms (p= 0.035) and in relation to S (p= 0.017) in the remineralization experiment. Other microbiological parameters were not significantly influenced by the treatments. Taking xylitol mints during short periods of time did not show advantages regarding the decrease of enamel demineralization or the promotion of enamel remineralization when compared to taking sorbitol mints. However, it did expressively alter the bacterial ecology of the biofilm, reducing the percentage and count of important groups of cariogenic microorganisms.

Made available in DSpace on 2014-06-11T19:31:03Z (GMT). No. of bitstreams: 0 Previous issue date: 2009-07-03Bitstream added on 2014-06-13T20:21:44Z : No. of bitstreams: 1 ernandes_fmpg_dr_sjrp.pdf: 1963066 bytes, checksum: 42dcae2ecfe9dfb358788c6490a13653 (MD5) / Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) / O principal produto da fermentação de açúcares por Zymomonas mobilis é o etanol quando glicose e frutose são utilizadas como fontes de carbono. Entretanto, quando sacarose é empregada na fermentação, o rendimento do etanol diminui devido à formação de subprodutos como levana, sorbitol, acetaldeído, ácido acético, pequenas quantidades de alguns álcoois superiores e fenol. A utilização de produtos agroindustriais, como o caldo e melaço de cana-deaçúcar, é uma alternativa para reduzir o custo final dos produtos de fermentação devido à disponibilidade de aquisição e composição química desses substratos. Este trabalho teve como objetivo utilizar substratos alternativos e otimizar as condições de fermentação para a produção de etanol, levana e sorbitol por Zymomonas mobilis CCT 4494. Também foi considerado o efeito da variação do substrato e de sais minerais adicionados nos meios de produção (sintético, caldo e melaços de cana-de-açúcar). Para a obtenção dos produtos de fermentação, foi aplicada a metodologia de superfície de resposta, seguindo um planejamento fatorial do tipo 27-2, de acordo com o modelo proposto por Box e Hunter, onde as variáveis independentes estudadas foram: pH inicial do meio de cultivo, temperatura de incubação, concentração do substrato e efeito da adição de KCl, K2SO4, MgSO4, CaCl2. Durante a realização das fermentações foi observado que a bactéria Zymomonas mobilis CCT 4494 se adaptou nos meios de fermentação contendo altas concentrações de sacarose e suportou a variação do pH e da temperatura de fermentação. O aumento da concentração da fonte de carbono favoreceu a formação dos produtos levana e etanol, entretanto, não houve produção de sorbitol. O meio sintético proporcionou maior rendimento de levana e etanol, enquanto que, os meios alternativos caldo e melaços de cana-de-açúcar... / The main product from fermentation of sugars by Zymomonas mobilis is ethanol when glucose and fructose are used as carbon sources. However, when sucrose is used in the fermentation medium, ethanol yield decreases due to the formation of by-products such as levan, sorbitol, acetaldehyde, acetic acid, small amounts of some superior alcohols and phenol. The use of agro industrial by products, such as sugarcane juice and molasses, is an alternative to reduce the final cost of fermentation products due to the constant availability and to the chemical composition of these by substrates. This study had the aim of using alternative substrates and of optimizing fermentation conditions for the production of ethanol, levan and sorbitol by Zymomonas mobilis CCT 4494. The effect of variation of substrate and mineral salts added to the production media (synthetic, sugarcane juice and molasses) was also considered. To obtain the fermentation products, response surface methodology was employed, following a 27-2 factorial planning, according to the model proposed by Box and Hunter, where the independent variables studied were: initial medium pH, incubation temperature, substrate concentration and effect of the addition of KCl, K2SO4, MgSO4, CaCl2. During the fermentations, it was noted that the bacteria Zymomonas mobilis CCT 4494 well adapted in the media containing high concentrations of sucrose and tolerated pH and temperature variations. The increase of carbon source concentration favored the formation of levan and ethanol, however, there was no sorbitol production. The synthetic medium offered higher levan and ethanol yield, whereas alternative media sugarcane juice and molasses, favored cellular growth. Among the independent variables analyzed with the best medium (synthetic) for biosynthesis of the biopolymer and ethanol, the ones that significantly (p<0.05) affected were KCl, K2SO4, CaCl2... (Complete abstract click electronic access below)

Apesar dos conhecimentos já existentes sobre o xilitol, seu efeito no desenvolvimento da cárie dentária ainda requer maiores esclarecimentos. Assim, este estudo in situ randomizado, duplo-cego e cruzado teve como objetivo determinar o efeito de xilitol em pastilhas na composição do biofilme dental e nos processos de desmineralização e remineralização do esmalte, comparado a um controle com pastilhas contendo sorbitol. Em duas fases de 14 dias, 11 voluntários utilizaram dispositivos palatinos com seis blocos de esmalte humano (três hígidos para avaliação da desmineralização + três previamente desmineralizados para avaliação da remineralização) com dureza de superfície conhecida. Sacarose 20% foi gotejada oito vezes ao dia apenas sobre os blocos hígidos. Cinco minutos após cada gotejamento, os dispositivos eram recolocados na boca e os voluntários chupavam pastilha com xilitol 88,3% ou sorbitol 84,5%. A utilização das pastilhas iniciou-se uma semana antes de cada fase experimental. Foi usado dentifrício sem flúor. Ao final de cada fase, o biofilme formado sobre os blocos foi coletado e dividido para análises microbiológica e bioquímica, e os fragmentos de esmalte foram analisados quanto a variação de dureza de superfície antes e após o experimento. A porcentagem de perda de dureza de superfície dos blocos de esmalte do grupo do xilitol apresentou uma tendência a ser menor do que aquela observada no grupo do sorbitol, embora a diferença não tenha sido significante (p= 0,066; ANOVA). Também não houve diferença estatisticamente significante entre os grupos em relação à porcentagem de ganho de dureza de superfície e à maioria dos parâmetros bioquímicos do biofilme. As médias das concentrações de fósforo inorgânico e de polissacarídeo intracelular no biofilme no experimento de desmineralização foram significantemente menores (p< 0,001 e p= 0,007) no grupo do sorbitol. As porcentagens de estreptococos do grupo mutans (SM) em relação ao total de estreptococos (S) (p= 0,037) no experimento de desmineralização, assim como as contagens de SM (p= 0,035) e lactobacilos (p= 0,048), e as porcentagens de SM em relação ao total de microrganismos (p= 0,035) e em relação a S (p= 0,017) no experimento de remineralização foram significantemente menores no biofilme do grupo do xilitol. Os demais parâmetros microbiológicos não foram influenciados de maneira significante pelos tratamentos. O uso de pastilhas com xilitol por curtos períodos de tempo não apresentou vantagem nem em diminuir a desmineralização nem em favorecer a remineralização do esmalte em comparação ao uso de pastilhas com sorbitol, mas alterou a ecologia bacteriana do biofilme, reduzindo as contagens e as porcentagens de importantes grupos de microrganismos cariogênicos. / In spite of the existing knowledge about xylitol, its effect on the development of dental caries is still a subject that requires further clarifications. Thus, this randomized, double-blind, crossover in situ study aimed at assessing the effect of xylitol mints in the dental biofilm composition and in the processes of enamel demineralization and remineralization, in comparison to a control group using sorbitol mints. During the two phases of 14 days, 11 volunteers wore palatal appliances with six human enamel blocks (three intact ones for evaluation of demineralization and three demineralized ones for the evaluation of remineralization) of known superficial hardness. A solution of 20% sucrose was dripped only on the intact enamel blocks eight times per day. Five minutes after each dripping, the appliances were placed back inside the mouth and the volunteers took either an 88.3% xylitol mint or an 84.5% sorbitol mint. Both types of mints started being used by the volunteers one week before each experimental phase. Non-fluoridated toothpaste was used. At the end of each phase, the biofilm formed over the blocks was collected and divided for microbiological and biochemical analysis and the enamel fragments were evaluated in relation to surface hardness variation before and after the experiment. The percentage of surface hardness loss of the enamel blocks in the xylitol group tended to be lower than in those of the sorbitol group although this difference was not significant (p= 0.066; ANOVA). Moreover, there were no statistically significant differences between the study groups regarding the percentage of surface hardness gain or regarding the biochemical parameters of the biofilm. The average concentrations of inorganic phosphorus and of intracellular polysaccharides in the biofilm for the demineralization experiment were significantly lower (p< 0.001 and p= 0.007) in the sorbitol group. The following results were significantly lower in the xylitol group biofilm: percentages of mutans streptococci (SM) in relation to the total number of streptococci (S) (p= 0.037) in the demineralization experiment; SM (p= 0.035) and lactobacilli (p= 0.048) count; and the percentages of SM in relation to the total number of microorganisms (p= 0.035) and in relation to S (p= 0.017) in the remineralization experiment. Other microbiological parameters were not significantly influenced by the treatments. Taking xylitol mints during short periods of time did not show advantages regarding the decrease of enamel demineralization or the promotion of enamel remineralization when compared to taking sorbitol mints. However, it did expressively alter the bacterial ecology of the biofilm, reducing the percentage and count of important groups of cariogenic microorganisms.

Sorbitol, the primary photosynthate and translocated carbohydrate in apple (Malus x domestica Borkh.), is converted to fructose by SORBITOL DEHYDROGENASE (SDH; EC 1.1.1.14) which is active in apple fruit throughout fruit development. Apple fruit set and early development is very sensitive to carbohydrate availability, but details on carbohydrate metabolism during this phase are limited. The first objective of this work was to determine if SORBITOL DEHYDROGENASE, the primary enzyme responsible for metabolism of the major phloem-transported carbohydrate sorbitol, is present and active during apple fruit set and early development. The second objective of this work was to determine if SDH genes are differentially expressed and how their patterns of expression may relate to SDH activity in apple seed and cortex during early fruit development. Nine different genes encoding SDH were determined from analysis of a cDNA library and genomic-clones. Northern, Western and ELISA analyses showed that SDH transcripts and SDH protein were present in the fruit during the first 5 weeks after bloom and comprised 7 to 8 % of the total extractable protein. Whole fruit SDH activity was highest at 2 to 3 weeks after bloom in each of three cultivars, Lodi, Redchief Delicious and Fuji. Seed SDH activity was found to be much higher than cortex SDH activity per mg and g FW, and seed SDH activity contributed significantly to whole fruit SDH activity during the first five weeks of development after bloom. Five of the nine SDH genes present in apple genome were expressed in apple fruit (SDH1, SDH2, SDH3, SDH6, SDH9). Expression of SDH6 and SDH9 was seed-specific and expression of SDH2 was cortex-specific. Using 2D SDS-PAGE and Western analyses, SDH isomers with pI values 4.2, 4.8, 5.5 and 6.3 were found in seeds, and SDH isomers with pI values 5.5, 6.3, 7.3 and 8.3 were found in cortex. The present work is the first to show that SDH is differentially expressed and highly active in seed and cortex during early development. Thus, SDH during apple fruit set and early development may play a primary role in defining fruit sink activity.

O interesse na aplicação de embalagens ativas para conservação de alimentos tem aumentado, bem como uma maior demanda por substâncias antimicrobianas naturais, devido à maior consciência dos consumidores quanto aos potenciais riscos à saúde ocasionados pelo consumo de compostos sintéticos. Entre os aditivos naturais, a própolis, por suas propriedades antibacteriana, antioxidante e antifúngica, tem despertado o interesse dos pesquisadores. Portanto, o objetivo deste trabalho foi investigar o efeito da adição do extrato etanólico de própolis (EEP) em filmes à base de gelatina plastificados com citrato de acetiltributila (CA) ou sorbitol (S), nas propriedades funcionais (propriedades mecânicas, solubilidade, permeabilidade ao vapor de água, parâmetros de cor e opacidade) e a atividade antimicrobiana dos filmes contra Staphylococcus aureus. Para a produção do EEP, 30g de resina de própolis (tipo 12) foram misturadas com 100mL de álcool etílico 80%, e a solução foi mantida sob agitação mecânica (500rpm, 30minutos) a 50ºC, e filtrada após 24horas sob refrigeração. Os filmes foram produzidos por casting com 2g de gelatina/100g de solução filmogênica, 30g de CA ou S/100g de gelatina, 35g de lecitina/100g de plastificante e o EEP nas concentrações de 0, 5, 40 ou 200g/100g de gelatina, e avaliados quanto às suas propriedades mecânicas (tração e perfuração), solubilidade em água (Sol), cor e opacidade, permeabilidade ao vapor de água (PVA), espectroscopia de infravermelho com transformada de Fourier (FTIR) e microscopia eletrônica de varredura, após acondicionamento (5 dias, UR = 58%, T = 25ºC). A atividade antimicrobiana dos filmes contra Staphylococcus aureus foi analisada pelo método da difusão em ágar. A incorporação de EEP causou variação na tensão (T) dos filmes com CA (58,7 a 66,4MPa), mas afetou mais intensamente os filmes com S (31,7 a 50,4MPa). Quanto à elongação (%), os filmes (CA e S) também apresentaram variações. Na perfuração, verificou-se que a adição de EEP não afetou significativamente a força máxima para ambos os plastificantes. Para a solubilidade em água, o filme com CA apresentou aumento significativo (Sol = 18,6%) para 200% de EEP, contudo, os filmes com S praticamente não variaram. Quanto à cor, para ambos os plastificantes estudados (CA e S) o aumento da concentração de EEP promoveu alterações significativas de L*, a*, b*, &Delta;E* e opacidade. As análises de PVA dos filmes (CA e S) mostraram que o aumento na concentração de EEP aumentou a propriedade de barreira dos filmes. Segundo a análise de FTIR, os filmes aditivados não apresentaram mudanças estruturais em relação ao controle, o que sugere que o EEP encontra-se disperso na matriz. As micrografias mostraram que a adição de EEP causou alterações, principalmente, nos filmes plastificados com S. A atividade contra S. aureus foi observada para os filmes (CA e S) com 40 e 200% de EEP, com aumento significativo do diâmetro de inibição em função do aumento da concentração de EEP. Os filmes mantiveram sua atividade antimicrobiana durante 177 dias de armazenamento. Estes resultados demonstraram a capacidade antimicrobiana dos filmes de gelatina aditivados com própolis, bem como sua estabilidade em função do tempo. / The interest in the use of active packaging for food conservation has increased, along with a higher demand for natural antimicrobial substances, due to higher consumer awareness of the potential health risks caused by synthetic compounds consumption. Among natural additives, propolis, has awaken the interest of the researchers for its antibacterial, antioxidant and antifungal properties. Thus, the aim of this work was to investigate the effect of the addition of an ethanolic propolis extract (EPE) to gelatin-based films plasticized with acetyltributyl citrate (AC) or sorbitol (S), on the functional properties (mechanical properties, solubility, water vapor permeability, color parameters and opacity) and the antimicrobial activity of the films against Staphylococcus aureus. For the EPE production, 30g for the propolis resin (type 12) were mixed with 100mL ethyl alcohol (80%), and the solution was maintained under mechanical stirring (500rpm, 30minutes) at 50°C, and filtered after 24hours under refrigeration. The films were produced by casting, using 2g of gelatin/100g of film forming solution, 30g of AC or S/100 g of gelatin, 35g of lecithin/100g of plasticizer and EPE in the concentrations of 0, 5, 40 or 200g/100g of gelatin, and evaluated for their mechanical properties (traction and puncture tests), water solubility (Sol), color and opacity, water vapor permeability (WVP), Fourier transform infrared spectroscopy (FTIR) and scanning electron microscopy, after storage (5 days, relative humidity = 58%, temperature = 25°C). The antimicrobial activity of the films against Staphylococcus aureus was analyzed by agar diffusion method. The EPE addition caused variation of the tensile strength (T) of the films with AC (58.7 a 66.4MPa), but affected more intensely the films with S (31.7 a 50.4MPa). The films (AC and S) also presented variations for the elongation (%). For the puncture force, the addition did not affect significantly the maximum force for any of the plasticizers. For the water solubility, the films with AC presented significant increase (Sol = 18,6%) for 200% of EPE, however the films with S did not vary. For both plasticizers studied (AC and S), the increase of EPE concentration promoted significant changes of the color parameters, L*, a*, b*, &Delta;E* and opacity. The WVP analysis of the films (AC and S) showed that the increase of the EPE concentration increased the barrier property of the films. According to FTIR analysis, the EPE addition in the films did not present structural changes compared to the control, suggesting that the EPE was disperse in the protein matrix. The micrographs showed that the EPE addition caused changes mainly in films plasticized with S. The activity against S. aureus was observed for the films containing 40 and 200% of EPE, with significant increase of inhibition diameter with increase of EPE concentration. The films kept their antimicrobial activities during 177 days of storage. These results demonstrated the antimicrobial capacity of gelatin-based films added of propolis, and their stability in time.