O uso indiscriminado dos plásticos tem gerado graves problemas ambientais de acúmulo de material na natureza, poluição das águas, além de ser proveniente do petróleo, um recurso não renovável. Isso tem causado preocupação e, portanto, os cientistas e indústrias vêm buscando alternativas ao uso desses materiais, como por exemplo o emprego de materiais naturais e/ou biodegradáveis e o aproveitamento de subprodutos desperdiçados. O amido é um polímero natural que tem aspectos que o tornam promissor, como sua biodegradabilidade, baixo custo e disponibilidade. Para prolongar a vida útil dos alimentos, investigou-se o amido como matéria prima para a produção de filmes biodegradáveis e revestimentos comestíveis ativos. Além de atuar como barreira a gases e como proteção mecânica, as embalagens também podem carrear substâncias ativas como antioxidantes, antimicrobianos, entre outros. Foi estudado o efeito da incorporação de (0,3; 0,5 e 1,0) g de α-tocoferol/100 g de dispersão filmogênica nas propriedades morfológicas, mecânicas, ópticas, de superfície, de barreira ao vapor d\'água e à luz UV/Visível e antioxidantes dos filmes de amido de semente de jaca. A adição de α-tocoferol aumentou a resistência à tração e o módulo de elasticidade nos filmes com até 0,5% de α-tocoferol. Também aumentou a barreira ao vapor d\'água, possivelmente devido à natureza hidrofóbica do α-tocoferol e ao efeito de tortuosidade causado por ele na matriz polimérica, embora a solubilidade em água não tenha diferido significativamente entre os filmes. O α-tocoferol proporcionou aumento na barreira contra a luz UV, especialmente nos comprimentos de onda entre (200 e 310) nm. A opacidade e o parâmetro de cor b* aumentaram significativamente, o que significa que a incorporação de α-tocoferol tornou os filmes mais opacos e amarelados. A hidrofobicidade da superfície aumentou significativamente (p < 0,05) com adição de α-tocoferol, principalmente no filme com 0,5% de α-tocoferol. A capacidade antioxidante aumentou significativamente com adição de até 0,5% de α-tocoferol. O filme controle (sem α-tocoferol) não apresentou capacidade antioxidante. Concluiu-se que o material de amido de semente de jaca incorporado de α-tocoferol tem potencial para ser usado como revestimento ativo para alimentos com alto teor de gordura. Entre as formulações produzidas, a de 0,5% de α-tocoferol foi a melhor, porque apresentou atividade antioxidante tão boa quanto o filme 1,0% de α-tocoferol e propriedades mais interessantes, como a microestrutural, mecânica, óptica e de superfície. / Indiscriminate use of plastics has been generating environmental problems, like accumulation of materials, water pollution, as well as depleting a non-renewable resource (crude oil). This has been causing concern, therefore scientists and industries are seeking alternatives for these materials, for example, the employment of natural and/or biodegradable materials and the reuse of wasted byproducts. Starch is a natural polymer with many promising aspects, like its biodegradability, low cost and availability. Aiming at prolonging shelf-life of food products, starch has been investigated as a feedstock for development of edible active films and coatings. Besides acting as a barrier against gases and as mechanical protection, packages also may carry active substances (antioxidants, antimicrobials, etc.). The effects of the incorporation of (0.3; 0.5 and 1.0) g α-tocopherol/100 g filmogenic dispersion on the morphological, mechanical, optical, surface, water vapor permeability, barrier to UV/Visible light and antioxidant properties of jackfruit seed starch films were studied. Addition of α-tocopherol increased the tensile strength and the elastic modulus up to 0.5%. It also increased the water vapor barrier, possibly due to α-tocopherol\'s hydrophobic nature and because it caused a tortuosity effect in the polymer matrix, although the film solubility was not significantly affected. α-tocopherol improved the UV light barrier, especially in the wavelengths between (200 and 310) nm. The opacity and the color parameter b* increased significantly, which means that films with α-tocopherol became more opaque and yellowish. Surface hydrophobicity increased with the incorporation of α-tocopherol, especially for the 0.5% α-tocopherol film. The antioxidant activity increased significantly up to 0.5% α-tocopherol. The film without α-tocopherol presented no antioxidant capacity. Therefore, jackfruit seed starch films incorporated with α-tocopherol have potential to be used as edible active coatings for high fat content foods. Between the formulations produced, the one with 0.5% of α-tocopherol was the best, because it presented an antioxidant activity similar to the film with 1.0% of α-tocopherol and superior characteristics, like microstructural, mechanical, optical and surface properties.

Conselho Nacional de Desenvolvimento CientÃfico e TecnolÃgico / A microalga cianofÃcea Spirulina platensis tem sido comercializada e estudada devido ao seu alto valor nutricional, possuindo vitaminas, minerais, proteÃnas, Ãcidos graxos poli-insaturados, carotenÃides, ficobilinas e polissacarÃdeos com propriedades terapÃuticas. O objetivo do presente trabalho foi avaliar os teores de -caroteno e -tocoferol presentes na microalga S. platensis cultivada em laboratÃrio com diferentes fotoperÃodos. A microalga S. platensis foi cultivada em seis aquÃrios de 20 L sob quatro condiÃÃes diferentes de iluminaÃÃo. Um cultivo foi submetido a um fotoperÃodo de 8 horas de luz (L) e 16 de escuro (E), o segundo a 12 L e 12 E, o terceiro a 16 L e 8 E e o Ãltimo exposto a 24 h de iluminaÃÃo constante. Os aquÃrios foram dispostos lado a lado em duas incubadoras de madeira, com temperatura ambiente de mantida em 28  2ÂC com auxÃlio de um âcoolerâ (220 V) e iluminÃncia na superfÃcie da Ãgua em torno de 78,1 μ E cm-2 s-1 fornecida por duas lÃmpadas fluorescentes de 40 W. Os cultivos foram monitorados por espectrofotometria a 680 nm e pela contagem dos filamentos (tricomas) microalgais utilizando um microscÃpio Ãptico comum. A anÃlise simultÃnea dos carotenos e tocoferÃis foi realizada por cromatografia lÃquida de alta eficiÃncia em uma coluna Waters Spherisorb-Hichrom S5 ODS-2 (4,6 x 250 mm) com uma fase mÃvel de MeOH: THF (90:10, v/v), fluxo de 1,5 mL min-1 e detecÃÃo a 450 nm e 292 nm, respectivamente. A extraÃÃo de -caroteno foi realizada suspendendo-se 0,1 g de Spirulina liofilizada em 10 mL de metanol:Ãgua Milli-Q (90:10, v/v). Para a saponificaÃÃo dos lipÃdios, a mistura foi homogeneizada com 7% de hidrÃxido de potÃssio, levada ao banhomaria a 70ÂC por 30 min e submetida à partiÃÃo em n-hexano que foi evaporado. O resÃduo foi suspenso em 1 mL de metanol e 100 μL foram injetados manualmente, em duplicata, no sistema cromatogrÃfico. O -caroteno foi detectado na biomassa obtida de todos os cultivos e o -tocoferol apenas nos cultivos com fotoperÃodos de 8 L e 16 E e de 12 L e 12 E, enquanto o -tocoferol nÃo foi detectado. O teor de -caroteno variou de acordo com a intensidade luminosa e quando as culturas foram expostas a 24 h de iluminaÃÃo constante, atingiram mais rapidamente a produtividade mÃxima e apresentaram o maior teor de -caroteno / The microalga cyanophyceae Spirulina platensis has been commercialized and studied due to its high nutritional value. It has shown to be a rich source of a great variety of vitamins, minerals, proteins, polyunsaturated fatty acids, carotenoids, phycobilins, and polysaccharides that exhibit potential therapeutic properties. The aim of the present work was to evaluate the content of -carotene and -tocopherol in S. platensis cultivated in laboratory under different conditions of photoperiods. S. platensis was cultivated within six 20-liter reservoirs under four different illumination conditions. The first culture was grown under 8- hour light (L) and 16-hour dark (D) photoperiod; the second, 12 L and 12 D photoperiod; the third, 16 L and 8 D photoperiod, and the forth cultivation condition was constant illumination. The reservoirs were placed side by side in two wood chambers, where room temperature was kept at 28  2ÂC with a cooler (220 V). The water surface illumination of 78.1 μE cm-2 s-1 was supplied by two 40-Watt fluorescent lamps. The cultures were monitored by spectrometry at 680 nm and by counting microalga filaments (trichomes) under an optical microscope. The simultaneous analysis of carotenoids and tocopherols was performed by high performance liquid chromatography in a Waters Spherisorb-Hichrom S5 ODS-2 (4.6 x 250 mm) column using MeOH:THF (90:10, v/v) as mobile phase, delivered at 1.5 mL min-1. The detector was set at 450 nm and 292 nm for carotenes and tocopherols, respectively. For the extraction of both compounds, three portions of 0.1 g lyophilized Spirulina were suspended in 10 mL methanol:MiliQ water (90:10, v/v). The mixture was homogenized with a 7% potassium hydroxide aqueous solution, brought to a water bath at 70ÂC for 30 min, for saponification, followed of partitioning into n-hexane which was evaporated to dryness. The residues were suspended in 1 mL methanol, and 100 μL aliquots were manually injected in the chromatographic system. -Carotene was detected in the biomass obtained from all the cultures and a-tocopherol was detected only in 8 L and 16 D and 12 L and 12 D photoperiods cultures. -Tocopherol, on the other hand, was not detected. The content of -carotene varied with illumination intensity and in the cultivation performed under constant illumination both maximum productivity and highest -carotene content were reached rapidly

O objetivo deste trabalho foi estudar o efeito da suplementação com vitamina E na alimentação de bovinos Nelore, sobre as características químicas e físico-químicas da carne desses animais. Esses estudos compreenderam: i) coleta de dados objetivos da cor de vários cortes cárneos para realização de um banco de dados; ii) estudo das características físico-químicas de cortes do contra filé (traseiro) e do peixinho (dianteiro) frescos e maturados por até 21 dias; iii) e verificação de possíveis alterações das características físico-químicas, inclusive da cor, de pedaços de contra filé e de carne (patinho) moída mantidas a –18ºC, durante 6 meses. Foram utilizados 24 novilhos Nelore, com 30 meses e peso médio de 279 kg, confinados, com dieta altamente energética, por 98 dias. 12 animais foram submetidos ao tratamento com 1000 mg de acetato de alfa-tocoferol, misturados a 100 g de farelo de milho, enquanto a outra metade recebeu apenas 100 g do farelo de milho. Os animais foram abatidos quando a espessura média da gordura atingiu média de 6 mm, medida por ultra-som, entre a 12ª e 13ª costelas. Coletaram-se então diversos cortes de carnes para a determinação das análises já descritas. No contexto atual das pesquisas, pode-se considerar que a suplementação com Vitamina E em bovinos Nelore não apresentou efeito positivo sobre as características químicas e físico-químicas da carne maturada por até 21 dias e congelada por até 6 meses de congelamento. / In general, the modern consumer uses to evaluate the meat quality through its attributes, such as, muscle color, thickness, tenderness and juiciness, among others. Withstanding the efforts done by Animal Science researchers for the last decades, meat quality can be lost during refrigerate or even frozen conservation due to the occurrence of fatty insaturated acid oxidation. An outlet to avoid this situation might be the use of vitamin E, as a diet supplement for animal feed. The objective of this work was to study the effects of diet supplementation with vitamin E on the physical and chemical characteristics of Nellore bovine meat. This study involved: i) objective data acquisition of color parameters of different muscle sections for data base storing; ii) study of the physical and chemical characteristics of the Longissimus dorsi and Quadriceps femuris muscles fresh and aged for 21 days; iii) and, verification of the possible modifications on the physical and chemical properties, or even color, of the Longissimus dorsi and Quadriceps femuris muscles grounded and kept at –18 ºC for 6 months. There were used 24 Nellore steers, with 30 months and 279 of mean live weight, confined and fed with highly energetic diets (73, 79 and 85 % of concentrate) for 98 days. Daily, half of the animals received 100 mg of acetate of alpha-tocopherol along with 100 g of corn meal, while the other animals were fed only with 100 g of corn meal. The animals were slaugtered when the backfat thickness, within the 12th and 13th ribs, measured by ultrasound, reached 6 mm. Boning and muscle sampling were done 24 hs after slaughter. During this period there were determined the color and pH of various fresh commercial cuts. There were collected 4 Longissimus dorsi and Supraspinatus steaks of every right half carcass, which were vacuum packed and aged for up to 21 days at 0 ºC. Another 7 steaks Quadriceps femuris were collected from the left half carcasses, packed and kept frozen for up to 6 months. There were also frozen in the same conditions, grounded samples of Quadriceps femuris collected from the left half carcass as well. The meat characteristics followed during aging and freezing were: the dry matter content, pH, water loss by exudation and cooking according to classical methods. The color of the samples was determined by a HunterLab colorimeter, using the CIELab color scale. The tenderness was determined using a WB probe with a texture analyzer TA.XT2, at a rate of 500 mm/min. The samples concentration of vitamin E and cholesterol were determined by High Performance Liquid Chromatography (HPLC). The results were statistically analyzed using the SAS software. In general, it was observed that in spite of the vitamin E concentration in the supplemented samples had been greater than from the control ones, it was not confirmed the protecting effect of this vitamin related to color or to texture of meat. The results indicated that at the acetate alpha-tocopherol concentration of the present study, vitamin E supplementation of Nellore bovine did not have influence on the physical and chemical characteristics of meat aged for 21 days and frozen for 6 months.

Orientador: Roberta Ceriani / Dissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Engenharia Química / Made available in DSpace on 2018-08-25T00:34:06Z (GMT). No. of bitstreams: 1 Damaceno_DanieladaSilva_M.pdf: 3241692 bytes, checksum: 461c874bf1a0705ce436bee9c205a2a1 (MD5) Previous issue date: 2014 / Resumo: Óleos e gorduras são uma fonte nutricional relevante de energia, de ácidos graxos essenciais, de vitaminas e de antioxidantes naturais, além da demanda recente como matéria-prima para produção de biodiesel. Em sua grande maioria, passam por algum tipo de processamento antes de ser destinado ao consumo humano. Além disso, derivados de óleos vegetais como ácidos graxos, ésteres graxos, triacilgliceróis, diacilgliceróis e monoacilgliceróis são importantes produtos oleoquímicos, de aplicação nas indústrias de alimentos e química. Neste contexto, o conhecimento e a descrição do comportamento de propriedades termofísicas de compostos graxos em função das diferentes variáveis de processamento podem ser de grande utilidade prática. No entanto, dados experimentais dessas propriedades na literatura aberta são bastante escassos. No caso da temperatura de ebulição de acilgliceróis parciais (mono- e diacilgliceróis), o banco de dados experimentais é formado por apenas seis valores para seis diferentes monoacilgliceróis a 0,13 kPa. Já para os tocoferóis, a literatura aberta apresenta um gráfico monolog da pressão de vapor (0,004 kPa a 0,5 kPa) em função da temperatura (453 K a 533 K). Nenhum dado pode ser encontrado para os diferentes tocoferóis (?-, ?-, ?- e ?-) Assim, nesse trabalho foram determinadas as temperaturas de ebulição de quatro acilgliceróis parciais, monocaprilina, monocaprina, dicaprilina e dicaprina a baixas pressões (1,0 kPa a 13,2 kPa) e dos tocoferóis ?-, ?-, ?- e ?- a 1,1 kPa, utilizando a técnica de calorimetria diferencial exploratória, que é bastante adequada para compostos susceptíveis à degradação térmica e oxidativa, e de custo bastante elevado (amostras na ordem de miligramas). Para os acilgliceróis parciais, parâmetros das equações de Clapeyron, Antoine e DIPPR (Design Institute for Physical Properties) foram ajustados. No caso da equação DIPPR, foram incluídos na regressão valores estimados pelo método de contribuição de grupos de Marrero e Gani para temperatura normal de ebulição e ponto crítico. A entalpia de vaporização foi estimada utilizando a relação de Clausius-Clayperon com a inclusão de um termo de correção para desvios a pressões mais elevadas. Já para a classe dos tocoferóis, uma equação DIPPR foi ajustada para a pressão de vapor e derivada para a entalpia de vaporização / Abstract: Oils and fats are an important source of energy, essential fatty acids, vitamins and nutritional source of natural antioxidants, plus the recent demand for feedstock for biodiesel production. Overwhelmingly, undergo some type of processing before being used for human consumption. Moreover, derived from vegetable oils such as fatty acids, fatty esters, trigacylglycerols, diacylglycerols and monoacylglycerols are important oleochemicals, application in the food and chemical industries. In this context, knowledge and description of the behavior of thermophysical properties of fatty due to different processing variables compounds may be of great practical utility. However, experimental data for these properties in the open literature are scarce. In the case of the boiling temperature of partial acylglcerols (mono- and diacylglycerols), the database of experimental data consists of only six values for six different monoacylglycerols at 0.13 kPa. As for the tocopherols, an open literature monolog shows a graph of vapor pressure (0.004 kPa up to 0.5 kPa) versus temperature (453 K up to 533 K). No data could be found for the different tocopherols (?-, ?-, ?- and ?-). Thus, in this work we determine the boiling temperatures four partial acylglycerols, monocaprin, monocaprylin, dicaprin and dicaprylin at low pressures (1.0 kPa up to 13.2 kPa) and ?-, ?-, ?- and ?- tocopherols 1.1 kPa, using the technique of differential scanning calorimetry, which is very suitable for compounds that are susceptible to thermal and oxidative degradation, and quite expensive (samples in the order of milligrams). For partial acylglycerols, parameters of Clapeyron, Antoine and DIPPR (Design Institute for Physical Properties) equations were adjusted. In the case of DIPPR equation, were included in the regression values estimated by the group contribution method of Marrero and Gani to the normal boiling point and critical point. The enthalpy of vaporization was estimated using the Clausius-Clapeyron relation with the inclusion of a correction term for deviations at higher pressures. As for the class of tocopherols a DIPPR equation was adjusted for vapor pressure and derived for the enthalpy vaporization / Mestrado / Desenvolvimento de Processos Químicos / Mestra em Engenharia Química

Leaves are responsible for the majority of the fixed carbon in most plant species. Along leaf development, the photosynthetic capacity increases until the organ reaches maturity. Consequently, at the onset of senescence the leaves have the highest photosynthetic activity, then, as the chloroplasts are dismantled and the photosynthetic machinery is degraded, leaves gradually lose the rate of carbon assimilation. Although the capacity to fix carbon declines as senescence progresses, nutrient remobilization from macromolecule degradation nourishes the developing sink organs. In this regard, delaying leaf senescence stands out as a promising strategy to increase plant yield as extends the window of time with maximum carbon fixation rate. Another approach that is receiving much attention is the manipulation of chlorophyll degradation once it potentially regulates photosynthetic capacity and affects the nutritional quality of harvestable organs. As chlorophyll is degraded, the released phytol is recycled and can be either stored (i.e. as fatty acid phytyl esters), used for chlorophyll synthesis or be incorporated in tocopherol biosynthesis. Tocopherols have high nutraceutical value due to their antioxidant properties. However, the majority of the studies regarding senescence and chlorophyll degradation were carried out in the model plant Arabidopsis thaliana or grasses, creating a knowledge gap about these processes in fleshy fruit-bearing plants of human diet interest. In this regard, the tomato, Solanum lycopersicum, is an excellent model not only for the genetic and genomic resources, but also for its agronomic and nutritional importance. Thus, this project aims to extend what is known about the effects of chlorophyll degradation and senescence manipulation over the metabolism and yield of tomato plants, as well as fruit nutritional quality. In order to evaluate the consequences of alteration in chlorophyll degradation, first the enzymes chlorophyllase and pheophytinase, both capable of dephytylating the chlorophyll molecule, were identified and characterised. An extensive phylogenetic, evolutive and transcriptional analysis allowed the identification of two groups of chlorophyllases, one putatively involved in the response to different stimuli, while the other may act in chlorophyll homeostasis. As for pheophytinase, only one group was identified, being related to physiologically programmed processes that trigger chlorophyll degradation (i.e. leaf senescence and fruit ripening). Given this scenario, pheophytinase was chosen to be constitutively knocked-down in order to evaluate the effects over the metabolism of leaves and fruits. As consequence of this manipulation, transgenic plants were impaired in the leaf senescence-associated chlorophyll breakdown, but, although with an initial delay, fruit ripening-associated degreening was not compromised. Several photosynthetic and biochemical parameters were signs of photoinhibition, possibly due to a deficiency in chlorophyll recycling in leaves. This led to an increase in sugar exportation towards fruits, ultimately increasing soluble sugar content of ripe fruits. However, as a consequence, carotenoid levels were reduced, what, at least partially, it was compensated by an increase in tocopherol content. The results indicated that pheophytinase plays a role beyond senescence-associated chlorophyll degradation and its manipulation led to the development of fruit with increased soluble sugars and tocopherols at the cost of lowering carotenoid levels. Thus, these evidences support the manipulation of chlorophyll breakdown as a strategy for improvement of fleshy fruit plants. In order to address the effects of senescence over yield and fruit quality, the transcription factor ORESARA1, which has been widely characterised in A. thaliana and is considered a key regulator of senescence initiation, was targeted. After a comprehensive phylogenetic analysis and the characterization of the regulatory mechanisms, one putative ortholog was selected to be silenced. As consequence of this manipulation, leaves displayed increased chlorophyll content. Moreover, as senescence was delayed, the extent of photosynthetic activity of the leaves was also expanded. As the number of fruits was increased in the knockdown lines, this reflected in an increment in the harvest index. Ripe fruits accumulated more soluble sugars and tocopherols. Collectively, the results support the manipulation of leaf senescence as a strategy for tomato yield improvement. Altogether, data obtained enhance the knowledge about the impacts of chlorophyll degradation and leaf senescence over the metabolism of fleshy-fruit plants, providing strategies to increase yield and nutritional quality of fruits / As folhas, para a maioria das espécies vegetais, são o principal órgão responsável pela fixação de carbono. Durante o desenvolvimento foliar, o potencial fotossintético aumenta até a folha atingir a sua maturidade. Consequentemente, no momento que o programa de senescência se inicia, a folha apresenta a maior taxa de fotossíntese, a qual passa então a declinar conforme o cloroplasto se desorganiza e a maquinaria fotossintética é degradada. Apesar da redução na fixação de carbono, o catabolismo de macromoléculas possibilita a remobilização de nutrientes para os órgãos dreno em desenvolvimento. Neste contexto, atrasar a senescência destaca-se como uma promissora estratégia para aumento da produtividade, uma vez que estende o período de máxima fixação de carbono das folhas. Outra estratégia que tem recebido atenção por, potencialmente, regular a capacidade fotossintética e afetar a qualidade nutricional dos órgãos coletáveis é a manipulação da degradação da clorofila. Durante o catabolismo deste pigmento, o fitol liberado é reciclado podendo ser armazenado (i.e. na forma de ésteres de fitil com ácidos graxos), ser utilizado na síntese de novas moléculas de clorofila ou ser incorporado na rota biossintética de tocoferóis. Estes últimos compostos, por seu potencial antioxidante, possuem alto valor nutracêutico. No entanto, a maior parte dos estudos sobre senescência e degradação de clorofila foi realizada na planta modelo Arabidopsis thaliana ou em gramíneas, tornando escassas as informações relativas a plantas com frutos carnosos de interesse para a dieta humana. Nesse âmbito, o tomateiro, Solanum lycopersicum, é um excelente modelo de estudo não apenas pela disponibilidade de recursos genético e genômicos, mas também pela importância agronômica e nutricional desta espécie. Assim, este trabalho pretende expandir o conhecimento acerca dos efeitos da manipulação da degradação de clorofila e da senescência sobre o metabolismo e produtividade do tomateiro, bem como sobre a qualidade nutricional dos frutos. De modo a se avaliar as consequências de alterações na degradação de clorofila, iniciou-se por identificar e caracterizar em tomateiro as enzimas clorofilase e feofitinase, as quais catalisam a defitilação da molécula de clorofila. Uma vasta análise filogenética, evolutiva e transcricional permitiu a identificação de dois grupos de clorofilases, um dos quais estaria envolvido na plasticidade de respostas a estímulos e o outro na homeostase dos níveis de clorofila. Já para feofitinase, somente um grupo foi identificado, o qual está relacionado a processos fisiologicamente programados que levam à degradação de clorofila (i.e. senescência foliar e amadurecimento de frutos). Dado o panorama obtido, a feofitinase foi escolhida para ser constitutivamente silenciada de modo a se avaliar as consequências para o metabolismo de folhas e frutos. Como consequência do silenciamento, as linhagens transgênicas mostraram-se incapazes de degradar clorofila durante a senescência, mas, embora com um atraso nas etapas iniciais, a degradação ao longo do amadurecimento de frutos não foi comprometida. Diversos parâmetros fotossintéticos e bioquímicos foram indicativos de fotoinibição, possivelmente em virtude de uma deficiência na reciclagem da clorofila em folhas. Isto acarretou em um aumento na exportação de açúcares para frutos, incrementando a concentração de açúcares solúveis nos frutos maduros, que, em contrapartida, resultou na queda nos teores de carotenoides. A queda nestes compostos antioxidantes foi, ao menos parcialmente, compensada por um aumento nos níveis de tocoferóis. Os resultados indicaram que a feofitinase possui um papel além da degradação de clorofila associada à senescência, e que sua manipulação leva ao desenvolvimento de frutos com maior teor de açúcares solúveis e de tocoferóis ao custo da redução no de carotenoides. Desta forma, estas evidências suportam a manipulação da clorofila como estratégia para o melhoramento de frutos carnosos. Para investigar o efeito da senescência sobre a produtividade e qualidade de frutos foi escolhido o fator de transcrição ORESARA1, o qual está amplamente caracterizado em A. thaliana e é considerado um regulador chave no desencadeamento deste processo. A partir de uma extensa análise filogenética e da caracterização de sua regulação, um putativo ortólogo foi selecionado como alvo para silenciamento. Como consequência desta manipulação, folhas apresentaram os níveis de clorofila incrementados. Além disto, taxas fotossintéticas maiores que as do genótipo controle foram mantidas por maior tempo indicando que a iniciação da senescência foi retardada. Assim, estas plantas produziram um maior número de frutos, consequentemente, aumentando o índice de colheita dessas linhagens. Os frutos maduros apresentaram maiores teores de açúcares solúveis e de tocoferóis. Os resultados demostraram que o retardo do início da senescência é uma estratégia efetiva para aumento da produtividade de tomateiro. Coletivamente, os resultados obtidos aprofundam o conhecimento acerta dos impactos da degradação de clorofila e senescência sobre o metabolismo de plantas com frutos carnoso, além de prover estratégias para se incrementar a produtividade e a qualidade nutricional de frutos

The Master’s thesis is focused on the analysis of biologically active substances in apricot kernel oil. Apricot oil has recently begun to be widely used in the cosmetics industry, particularly as a part of massage oils or as part of moisturizing creams. The aim of this thesis is to characterize and assess the quality of apricot oil, determination of the content of selected active ingredients, through a series of analytical methods, and subsequent use of this oil in cosmetics and food industry. The thesis is based on the comparison of the quality of pure apricot oil and the oil after the fermentation process. It has been found that apricot kernel oil actually contains large amounts of unsaturated fatty acids. The excellent antioxidant activity of apricot kernels was proven too, especially those after fermentation. In addition to antioxidant activity, the samples contain a relatively large amount of tocopherol after the fermentation. Tocopherol is important for the uptake of free radicals in lipid bilayers, thus slowing down aging, protect the membranes and also prevent from possible cardiovascular and oncological diseases. Furthermore, in the Master’s thesis there were confirmed not only the hydration effects of creams with apricot oil, but also their ability of inhibic transepidermal loss of water from the epidermis. Creams were tested on five probands, aged 23–24, with different skin types. After the application of creams to their skin, these properties greatly improved, the skin has been regenerated and hydrated for 24 hours after the treatment.

Einleitung Adipositas ist ein zunehmendes Problem bei Menschen und Haustieren, z.B. in Pferden. Ponyrassen sind dabei besonders prädisponiert, wobei die Gründe bisher nicht abschließend geklärt werden konnten. Humane Adipositas geht mit einer fettigen Infiltration der Leber einher, die sogenannte Non-Alcoholic Fatty Liver Disease, welche zu einer hepatozellulären Entzündung führt. Es ist bisher nicht bekannt, ob Adipositas in Equiden auch zu hepatischen Veränderungen führt. Menschliche Fettleibigkeit ist zusätzlich mit systemischer Entzündung und gesteigertem oxidativen Stress verbunden. Das führte zu intensiven Untersuchungen von anti-inflammatorischen und antioxidativen Faktoren (z.B. Vitamin A - Retinol und Vitamin E - α-Tocopherol) in der humanen Adipositas Forschung. Viele Studien konnten ein Absinken von Vitamin A und Vitamin E in fettleibigen Menschen feststellen. Ziele Die vorliegende Studie sollte den Einfluss von zunehmendem Körpergewicht (KG) in Ponys und Pferden auf mehrere Parameter untersuchen: (1) Serum Leberenzymaktivitäten und Serum Gallensäuren (GS), (2) Leberfettgehalt, (3) hepatische messenger Ribonukleinsäure (mRNA) Level von Entzündungsmarkern und Markern des Lipidmetabolismus und (4) Serum Konzentrationen von Retinol und α-Tocopherol. Zusätzlich sollten Ponys und Pferde im Verlauf dieser Studie verglichen werden, um eventuelle Gründe für die Rasseprädisposition der Ponys für metabolische Störungen zu identifizieren. Material und Methoden Zehn Shetland Ponys und 9 Warmblut Pferde, die initial nicht adipös waren, wurden über 2 Jahre mit 200% des Erhaltungsbedarfes für umsetzbare Energie gefüttert. Die Entwicklung des KG, des Body Condition Scores (BCS) und des Cresty Neck Scores (CNS) wurde wöchentlich erfasst. Während der Fütterungsphase wurde zu 6 Zeitpunkten (ZP) Blut für die Bestimmung von Serum Leberenzymaktivitäten (Alkaline Phosphatase (ALP), Aspartat Aminotransferase (AST), Glutamat Dehydrogenase (GLDH), Gamma-Glutamyl Transferase (GGT)) und Serum GS entnommen und zu 7 ZP wurde Blut für die Analyse von Serum Retinol und α-Tocopherol gewonnen. An 3 ZP wurde durch Laparotomie Lebergewebe in Vollnarkose entnommen. Die Leberbiopsien wurden histologisch auf ihren Fettgehalt untersucht und mittels quantitativer Echtzeit Polymerase-Kettenreaktion (RT-qPCR) wurden die mRNA Level von Entzündungsmarkern (Nuclear Factor-κB (NF-κB), Interleukin-1β (IL-1β), IL-6, Tumor Nekrose Faktor α (TNFα), Differenzierungsgruppe 68 (CD68), Chemerin) und Lipid Metabolismus Markern (Lipoprotein Lipase (LPL), Fettsäuren Bindungsprotein 1 (FABP1) bestimmt. Die Daten wurden mittels statistischem Software Programm ausgewertet (STATISTICA, version 12, StatSoft GmbH, Hamburg, Deutschland). Nach Prüfung auf Normalverteilung der Daten, wurden geeignete statistische Tests angewendet mit einem statistischen Signifikanzniveau bei P < 0,05. Die Tierschutzkommission des Bezirks Leipzig genehmigte das Projekt in Übereinstimmung mit deutschen Rechtsvorschriften (Nr. TVV 32/15). Ergebnisse Ponys und Pferde zeigten einen signifikanten Anstieg von KG (Mittelwert ± SD; Ponys: 29,9 ± 19,4%; Pferde: 17 ± 6,74%), BCS (Median (25./75. Perzentil); Ponys: 157% (115/349); Pferde: 142% (128/192)) und CNS (Median (25./75. Perzentil); Ponys: 165% (123/500); Pferde: 200% (160/225)) induziert durch die hyperkalorische Fütterung über 2 Jahre. Das ansteigende KG hat keine Steatosis in der Mehrheit der Equiden ausgelöst. Die mRNA Level von IL-6, TNFα, CD68 und IL-1β in der Leber wurden nicht beeinflusst. Die Leber mRNA Level von Chemerin sind signifikant angestiegen in Ponys (x-facher Anstieg: 1,89) und Pferden (x-facher Anstieg: 2,04). Signifikante Unterschiede zwischen den Rassen hinsichtlich der Serum GLDH Aktivitäten, Serum GS Konzentrationen und der hepatischen mRNA LPL Level konnten festgestellt werden. Die Serum α-Tocopherol Konzentrationen stiegen in Ponys und Pferden signifikant an und korrelierten positiv mit der Vitamin E Aufnahme. Die Serum Retinol Konzentrationen fluktuierten während der Studie, ohne mit der Aufnahme zu korrelieren. Schlussfolgerungen Frühe Fettleibigkeit in Equiden führt nicht zwangsläufig zu einer Steatose mit hepatozellulärer Entzündung. Gemäß der Hypothese zeigten Ponys und Pferde allerdings unterschiedliche hepatische Reaktionsmuster nach KG Zunahme. Das könnte die höhere Empfänglichkeit von Ponys für metabolische Erkrankungen erklären. Chemerin konnte als interessanter Marker für die equine Adipositas Forschung identifiziert werden. Serum Konzentrationen von Retinol und α-Tocopherol wurden durch die KG Zunahme nicht beeinflusst.

BACKGROUND: α-tocopherol (AT) and γ-tocotrienol (GT3) are vitamin E isoforms considered to have potential chemopreventive properties. AT has been widely studied in vitro and in clinical trials with mixed results. The latest clinical study (SELECT trial) tested AT in prostate cancer patients, determined that AT provided no benefit, and could promote cancer. Conversely, GT3 has shown antineoplastic properties in several in vitro studies, with no clinical studies published to date. GT3 causes apoptosis via upregulation of the JNK pathway; however, inhibition results in a partial block of cell death. We compared side by side the mechanistic differences in these cells in response to AT and GT3. METHODS: The effects of GT3 and AT were studied on androgen sensitive LNCaP and androgen independent PC-3 prostate cancer cells. Their cytotoxic effects were analyzed via MTT and confirmed by metabolic assays measuring ATP. Cellular pathways were studied by immunoblot. Quantitative analysis and the determination of relationships between cell signaling events were analyzed for both agents tested. Non-cancerous prostate RWPE-1 cells were also included as a control. RESULTS: The RAF/RAS/ERK pathway was significantly activated by GT3 in LNCaP and PC-3 cells but not by AT. This activation is essential for the apoptotic affect by GT3 as demonstrated the complete inhibition of apoptosis by MEK1 inhibitor U0126. Phospho-c-JUN was upregulated by GT3 but not AT. No changes were observed on AKT for either agent, and no release of cytochrome c into the cytoplasm was detected. Caspases 9 and 3 were efficiently activated by GT3 on both cell lines irrespective of androgen sensitivity, but not in cells dosed with AT. Cell viability of non-cancerous RWPE-1 cells was affected neither by GT3 nor AT. CONCLUSIONS: c-JUN is a recognized master regulator of apoptosis as shown previously in prostate cancer. However, the mechanism of action of GT3 in these cells also include a significant activation of ERK which is essential for the apoptotic effect of GT3. The activation of both, ERK and c-JUN, is required for apoptosis and may suggest a relevant step in ensuring circumvention of mechanisms of resistance related to the constitutive activation of MEK1.

In cystic fibrosis (CF), pancreatic insufficiency and a diminished bile acid pool cause malabsorption of important nutrients and dietary components leading to deficiency, poor nutritional status, and oxidative stress. Of particular significance is the malabsorption of fat-soluble nutrients and antioxidants, which are important for normal immune and neurologic function. Patients with CF often are deficient in these compounds despite supplementation with the current standard of care therapy. The objective was to compare the pharmacokinetic profile of this water-soluble vitamin E formulation (Aqua-E) with an oil-based softgel formulation in a malabsorbing patient population. Patients with CF who had documented malabsorption were recruited for participation in this pharmacokinetic study. Patients who met inclusion and exclusion criteria discontinued vitamin E supplementation, except for that in a multivitamin, for 7 to 21 days before the day of dosing. Patients were randomized to a single dose of 20 ml of Aqua-E or three oil-based softgels, which contained equivalent amounts of tocopherols. Blood was drawn from patients at time 0, 2, 4, 8, 24, 48, and 168 hr and analyzed for tocopherols. Eight patients were enrolled in the study and randomized to Aqua-E or softgels. The primary outcome, the absorption of γ-tocopherol in Aqua-E (AUC=115 μg/ml*hr), was significantly greater than that of oil-based softgels (AUC=25.3 μg/ml*hr; P=0.013). Total-tocopherols (α+γ +δ) in Aqua-E (AUC=294 μg/ml*hr) showed a strong trend toward increased absorption compared with that of oil-based softgels (AUC=117 μg/ml*hr; P=0.09). In conclusion, this novel, water-soluble formulation showed a marked and statistically significant increase in absorption of γ-tocopherol in malabsorbing patients with CF compared with an oil-based formulation.

With use of deuterium-labeled isotopes of RRR-and all-rac-α-tocopheryl acetate, the transport of vitamin E in pregnancy was evaluated to determine whether the placenta discriminates between these compounds. Fifteen pregnant subjects were recruited 5 d before delivery to receive 15, 30, 75, 150, or 300 mg vitamin E/d in capsules containing d3-RRR-α-tocopheryl acetate and d6-all-rac-α-tocopheryl acetate (1:1, by wt). Maternal blood was obtained before dosing, at hospital admission, and at parturition. Cord blood samples were obtained at parturition. Deuterium-labeled and unlabeled tocopherol contents were determined by gas chromatography-mass spectrometry in plasma and lipoproteins (chylomicrons, VLDL, LDL, and HDL). Maternal plasma and lipoproteins obtained at delivery had higher concentrations of d3-RRR-α- tocopherol than d6-all-rac-α-tocopherol regardless of the vitamin E dose administered (P < 0.05). Cord plasma at delivery also had higher concentrations of d3-RRR-α-tocopherol than d6-all-rac-α-tocopherol in plasma irrespective of the dose administered (P < 0.05). In lipoproteins isolated from cord blood, tocopherol concentrations were greatest in the HDL fraction (P < 0.05), whereas in maternal blood they were greatest in the LDL fraction (P < 0.05). We conclude that the placental-fetal unit, the fetal liver, or both further discriminate between RRR- and all-rac-α-tocopherol.