The Center for Disease Control and Prevention (CDC) reports indicate that 8% and 6.3% of Kentuckians consume enough fruits and vegetables, respectively. The Plate It Up! Kentucky Proud (PIUKP) project is a recipe-development project that aims to boost produce consumption by incorporating local fruits and vegetables. The purpose of this study was to implement promotional strategies using PIUKP recipes at farmers’ markets and determine their effects on consumers’ intent of purchasing and preparing the produce. The study was conducted at nine farmers’ markets across Kentucky (n=300) in collaboration with Cooperative Extension agents/assistants. The consumers’ impression of the sample was positively associated with their intent to purchase fruits and vegetables the same day (t = 0.36; p < 0.0001), in future (t=0.43; p < 0.0001), and prepare the respective recipes (t=0.51; p < 0.0001). Distribution of recipe cards was also positively correlated with consumers’ intent to prepare recipes (t=0.35; p < 0.0001). However, no significant association was found between the self-reported fruit and vegetable intake and their respective dermal carotenoid score. Findings from this study support the use of promotional strategies as a means to influence produce intake among farmers market consumers. Future studies can apply these strategies and explore the extent of effect they have on dietary intake.

<p>In photosynthesis, sunlight is converted into chemical energy that is stored mainly as carbohydrates and supplies basically all life on Earth with energy.</p><p>In order to efficiently absorb the light energy, plants have developed the outer light harvesting antenna, which is composed of ten different protein subunits (LHC) that bind chlorophyll a and b as well as different carotenoids. In addition to the light harvesting function, the antenna has the capacity to dissipate excess energy as heat (feedback de-excitation or qE), which is crucial to avoid oxidative damage under conditions of high excitation pressure. Another regulatory function in the antenna is the state transitions in which the distribution of the trimeric LHC II between photosystem I (PS I) and II is controlled. The same ten antenna proteins are conserved in all higher plants and based on evolutionary arguments this has led to the suggestion that each protein has a specific function.</p><p>I have investigated the functions of individual antenna proteins of PS II (Lhcb proteins) by antisense inhibition in the model plant Arabidopsis thaliana. Four antisense lines were obtained, in which the target proteins were reduced, in some cases beyond detection level, in other cases small amounts remained.</p><p>The results show that CP29 has a unique function as organising the antenna. CP26 can form trimers that substitute for Lhcb1 and Lhcb2 in the antenna structure, but the trimers that accumulate as a response to the lack of Lhcb1 and Lhcb2 cannot take over the LHC II function in state transitions. It has been argued that LHC II is essential for grana stacking, but antisense plants without Lhcb1 and Lhcb2 do form grana. Furthermore, LHC II is necessary to maintain growth rates in very low light.</p><p>Numerous biochemical evidences have suggested that CP29 and/or CP26 were crucial for feedback de-excitation. Analysis of two antisense lines each lacking one of these proteins clearly shows that there is no direct involvement of either CP29 or CP26 in this process. Investigation of the other antisense lines shows that no Lhcb protein is indispensable for qE. A model for feedback de-excitation is presented in which PsbS plays a major role.</p><p>The positions of the minor antenna proteins in the PS II supercomplex were established by comparisons of transmission electron micrographs of supercomplexes from the wild type and antisense plants.</p><p>A fitness experiment was conducted where the antisense plants were grown in the field and seed production was used to estimate the fitness of the different genotypes. Based on the results from this experiment it is concluded that each Lhcb protein is important, because all antisense lines show reduced fitness in the field.</p>

In photosynthesis, sunlight is converted into chemical energy that is stored mainly as carbohydrates and supplies basically all life on Earth with energy. In order to efficiently absorb the light energy, plants have developed the outer light harvesting antenna, which is composed of ten different protein subunits (LHC) that bind chlorophyll a and b as well as different carotenoids. In addition to the light harvesting function, the antenna has the capacity to dissipate excess energy as heat (feedback de-excitation or qE), which is crucial to avoid oxidative damage under conditions of high excitation pressure. Another regulatory function in the antenna is the state transitions in which the distribution of the trimeric LHC II between photosystem I (PS I) and II is controlled. The same ten antenna proteins are conserved in all higher plants and based on evolutionary arguments this has led to the suggestion that each protein has a specific function. I have investigated the functions of individual antenna proteins of PS II (Lhcb proteins) by antisense inhibition in the model plant Arabidopsis thaliana. Four antisense lines were obtained, in which the target proteins were reduced, in some cases beyond detection level, in other cases small amounts remained. The results show that CP29 has a unique function as organising the antenna. CP26 can form trimers that substitute for Lhcb1 and Lhcb2 in the antenna structure, but the trimers that accumulate as a response to the lack of Lhcb1 and Lhcb2 cannot take over the LHC II function in state transitions. It has been argued that LHC II is essential for grana stacking, but antisense plants without Lhcb1 and Lhcb2 do form grana. Furthermore, LHC II is necessary to maintain growth rates in very low light. Numerous biochemical evidences have suggested that CP29 and/or CP26 were crucial for feedback de-excitation. Analysis of two antisense lines each lacking one of these proteins clearly shows that there is no direct involvement of either CP29 or CP26 in this process. Investigation of the other antisense lines shows that no Lhcb protein is indispensable for qE. A model for feedback de-excitation is presented in which PsbS plays a major role. The positions of the minor antenna proteins in the PS II supercomplex were established by comparisons of transmission electron micrographs of supercomplexes from the wild type and antisense plants. A fitness experiment was conducted where the antisense plants were grown in the field and seed production was used to estimate the fitness of the different genotypes. Based on the results from this experiment it is concluded that each Lhcb protein is important, because all antisense lines show reduced fitness in the field.

O objetivo desta pesquisa foi estudar a formação de partículas de licopeno em pó de colágeno hidrolisado (CH) comercial usando CO2 supercrítico (CO2-SC). O estudo consistiu em três etapas: a medida de solubilidade do CH em CO2-SC por método estático para conhecer a afinidade do CH pelo CO2, a formação de partículas de misturas físicas compostas de diferentes proporções de CH/licopeno (4:1, 6:1, 8:1 e 10:1) através de processamento em uma autoclave com CO2-SC a 140 bar e 50 °C em agitação de 1.250 rpm por 45 min para definir a proporção de CH/licopeno, e o estudo do efeito de outras condições de CO2-SC (150 bar e 50 °C, 150 bar e 60 °C, 250 bar e 50 °C, 250 bar e 60 °C) na formação de partículas de licopeno em CH para identificar a melhor condição de CO2-SC. As partículas resultantes foram caracterizadas com relação à sua morfologia por microscopia eletrônica de varredura, distribuição de tamanho por difração a laser, comportamento térmico por calorimetria diferencial de varredura, estrutura química por espectroscopia de infravermelho de transformada de Fourier, liberação de carga, e estrutura física interna das partículas por microscopia de laser confocal para conhecer o mecanismo de formação das partículas pelo processamento com CO2-SC. Os resultados indicaram que o CH é pouco solúvel em CO2-SC, motivo pelo qual não há potencial para processos de micronização supercrítica relacionados à solubilidade do biomaterial carreador em CO2-SC. No estudo de proporções de CH/licopeno foi observado que o processo com CO2-SC a 140 bar e 50 °C permite a obtenção de partículas de licopeno em CH na forma de pó de coloração vermelha com intensidade dependente da quantidade de licopeno inicial, sem mudanças importantes na estrutura física porosa do CH, e um aumento de tamanho das partículas. Recomenda-se a proporção 10:1 de CH/licopeno, pois nela se evidenciou menor aglomeração de partículas e formação de fissuras na superfície das partículas que facilitam a incorporação do licopeno em CH seguindo um mecanismo de impregnação supercrítica propiciado pela sorção de CO2 e arraste simultâneo de licopeno ao interior da estrutura física das partículas de CH. Na exploração de outras condições de CO2-SC, na temperatura de 60 °C (150/250 bar) obteve-se uma massa de aparência viscoelástica sem utilidade para a formação de partículas. Entanto, na temperatura de 50 °C foi observado que as partículas de licopeno em CH formadas com CO2-SC a 150 bar, quando comparadas com as formadas com CO2-SC a 250 bar, evidenciaram formação de fissuras que resultaram na melhor dispersão e maior carga de licopeno na estrutura interna do CH. Além disso, as análises do comportamento térmico e o espectro de infravermelho destas partículas evidenciaram a formação de interações eletrostáticas entre o licopeno e o CH favorecidas pelo processamento com CO2-SC. Assim, conclui-se que o processamento com CO2-SC a 140/150 bar e 50 °C poderia ser utilizada para o design de partículas de licopeno impregnadas em CH, gerando um ingrediente com possível ampla atividade funcional pela atividade biológica do CH em tecidos conjuntivos e pela atividade antioxidante do licopeno. / The objective of this research was to study the formation of lycopene particles in commercial powder of hydrolyzed collagen (HC) using supercritical CO2 (SC-CO2). The study consisted of three steps: the solubility measurement of HC in SC-CO2 by static method to know the affinity of HC by CO2, the formation of particles of physical mixtures composed of different ratios of HC/lycopene (4:1, 6:1, 8:1 and 10:1) by autoclaving with SC-CO2 at 140 bar and 50 °C under stirring at 1,250 rpm for 45 min to define the HC/lycopene ratio, and the study of the effect of other conditions of SC-CO2 (150 bar and 50 °C, 150 bar and 60 °C, 250 bar and 50 °C, 250 bar and 60 °C) in the particle formation of lycopene in HC to identify the best SC-CO2 condition. The resulting particles were characterized with respect to their morphology by scanning electron microscopy, size distribution by laser diffraction, thermal behavior by differential scanning calorimetry, chemical structure by Fourier transform infrared spectroscopy, charge release, and physical structure of the particles by confocal laser microscopy to know the mechanism of particle formation by SC-CO2 processing. The results indicated that HC is poorly soluble in SC-CO2 and therefore has no potential for supercritical micronization processes related to the solubility of the biomaterial in SC-CO2. In the study of ratios of HC/lycopene it was observed that the process with SC-CO2 at 140 bar and 50 °C allowed to obtain lycopene particles in HC with powder appearance of red color with intensity depending on the amount of initial lycopene, without important changes in the porous physical structure of HC, and an increase in particle size. It is recommended a 10:1 ratio of HC/lycopene, since this ratio showed less particle agglomeration, and formation of fissures on the surface of the particles facilitating the incorporation of lycopene into HC following a mechanism of supercritical impregnation propitiated by CO2 sorption and simultaneous drag of lycopene to the interior of the physical structure of the HC particles. In the exploration of other conditions of SC-CO2, at 60 °C (150/250 bar) a mass of viscoelastic appearance was obtained without utility for the formation of particles. However, at 50 °C was observed that the lycopene particles in HC formed with SC-CO2 at 150 bar, when compared with the particles formed with SC-CO2 at 250 bar evidenced the formation of fissures, which had positive influence in the dispersion and charge of lycopene in the internal structure of HC. In addition, analyzes of the thermal behavior and the infrared spectrum of these particles evidenced the formation of electrostatic interactions between lycopene and HC favored by SC-CO2 processing. Thus, it can be concluded that SC-CO2 processing at 140/150 bar and 50 °C could be used for the design of lycopene particles impregnated in HC, producing an ingredient with possible broad functional activity due to the biological activity of HC in connective tissues and the antioxidant activity of lycopene.

Chez les mâles, la testostérone joue un rôle important dans l'expression des signaux sexuels attractifs pour la femelle. Cependant, de nombreuses études mettent en évidence l'effet négatif de la testostérone sur les défenses immunitaires ou la résistance aux parasites. Selon l'hypothèse du handicap immunitaire (ICHH, Folstad et Karter, 1992), ce compromis permettrait d'assurer l'honnêteté du signal sexuel ; les mauvais mâles ne pouvant pas exprimer de signaux sexuels de bonne qualité tout en gérant l'immunosuppression due à la testostérone. Ma thèse a pour objectif de définir le rôle de la testostérone dans l'honnêteté des signaux sexuels dans un contexte de communication multimodale. Elle permet également de comprendre les mécanismes physiologiques par lesquels la testostérone module les signaux sexuels acoustiques et visuels chez les mâles Hyla arborea, un amphibien connu pour utiliser ces deux types de signaux pour attirer les femelles. Nos résultats montrent que les signaux sexuels acoustiques et visuels chez cette espèce sont testostéronedépendants. En revanche, puisque la testostérone n'a pas d'effet immunosuppresseur dans notre étude, elle ne serait pas le médiateur direct de l'honnêteté des signaux chez cette espèce. D'un point de vue physiologique, l'effet de la testostérone sur le chant s'explique par l'amélioration des propriétés contractiles des muscles du tronc responsables du chant lié à une augmentation de l'efficacité mitochondriale. D'autre part, les analyses biochimiques montrent que la couleur du sac vocal varie linéairement avec la quantité de pigments caroténoïdes présents dans le plasma, qui pourrait être testostérone-dépendant. Enfin, cette thèse montre également que la testostérone agit sur le comportement de chasse permettant l'acquisition de ressources énergétiques et de caroténoïdes pour le chant et la couleur des mâles / In males, testosterone plays an important role on attractive sexual signal expression. However, numerous studies highlighted that testosterone also has negative effects on immunity defenses and parasite resistance capacity. According to the immunocompetence handicap hypothesis (ICHH, Folstad and Karter, 1 992), this trade-off assures signal honesty because only good males could express powerful signals while managing the immunosuppression of testosterone. My thesis aims to define the role of testosterone on the signal honesty in a multimodal communication context. Moreover it allows us to understand some physiological mechanisms involved in the modulation of acoustic and visual signals in males Hyla arborea, an amphibian known to use both acoustic and visual signals to attract females. Our results show that acoustic and visual signals are testosterone-dependent. In contrast, since testosterone has no immunosuppression effect in our study, it would not be the direct mediator of the signal honesty in this species. From a physiological point of view, the testosterone effects on calls could be explained through the positive effect of testosterone on trunk muscle mitochondrial efficacy and their contractile properties. Besides, biochemical analyses show that vocal sac coloration varies with the amount of carotenoid pigments present in the plasma that could be testosterone-dependent. Finally, this thesis also assumes that testosterone acts on the hunting behavior permitting the acquisition of energy resources and carotenoids for singing and coloration of males

Technique of pulsed field gel electrophoresis (PFGE) has found widespread use in the analysis of the genome of all life organisms. It is applied to the separation of the large DNA molecules above thousands base pairs up to millions of base pairs in size, where using conventional gel electrophoresis techniques are not possible (for instance large bacterial, yeast, fungal or mammalian chromosome). Presented work was realized as a comparative analysis of genome of several carotenogenic yeasts. The conditions of isolation and analysis of chromosomal yeast DNA were optimized. A lysis of yeast cells and deproteination of DNA within agarose chops was shown as the most appropriate method for DNA isolation. Cultivation to late exponential phase (50 hours) is the most suitable to obtaining intact DNA in sufficient amount and quality. Carotenogenic yeasts undergo the random mutagenesis using alkylation reagent ethyl methanesulfonate (EMS). Genome of pigment overproducing mutants was analyzed by pulsed field gel electrophoresis and amount of carotenoids by high pressure liquid chromatography (HPLC). However, overproduction of beta-carotene was analyzed in mutant strains Rhodotorula glutinis (10.6 g/l of biomass enriched 0,34 mg/g of beta-carotene) and Cystofilobasidium capitatum (8.5 g/l of biomass enriched 0,23 mg/g of beta-carotene). Selection of mutant strains overproducing carotenoid pigments was in presented experiment series successful in almost all analyzed strains except in the case of the strain Rhodotorula aurantiaca.

No description available.

No description available.

The effects of light, light "mimicking" chemicals, and protein synthesis inhibitors on the photo-induced carotenogenesis of Corynebacterium 7EIC were studied. Changes in the dosage of fluorescent light applied to dark grown cells showed a dose related carotenogenic response. Maintaining the same dosage but varying the wavelength of monochromatic light revealed that light with a wavelength of 280 to 450nm was responsible for photo-induction. It further showed a peak of photo-induction between the wavelengths of 370 and 430nm. The light "mimicking" chemicals antimycin A and p-Chloromercurybenzoate were shown to have no light "mimicking" effects. The transcriptional inhibitor of protein synthesis actinomycin D partially inhibited, and chloramphenicol a translational inhibitor, completely inhibited photo-induced carotenogenesis.

<p>Successful adoption of biofortified orange maize in developing countries requires careful consideration of factors across the chain from farm to fork. This includes consideration of post-harvest storage conditions optimal for the retention of both proviatamin A carotenoids and cooking quality critical to consumers. In these considerations, identification of economical storage methods is critical considering the limitations within specific countries that biofortified maize is being disseminated. To address these points, this dissertation research focused on evaluation of the utility of the Purdue Improved Crop Storage (PICS) bags as a post-harvest storage solution for biofortified maize. The specific focus of this research was to monitor retention of provitamin A and other carotenoids in two biofortified maize genotypes (OPVI and OPVII) as well as storage effect on flour functionality. Finally, a preliminary assessment of the impacts of storage on carotenoid bioaccessibility was completed to begin to translate findings to practice.</p><p>Maize grain from 2016 harvest was stored at ambient conditions for eight months in either PICS bags with or without an O₂ scavenger, (PICS-oxy) and (PICS-noxy), respectively and compared to storage in common polypropylene woven bags (control). After 4 months of storage carotenoid content was significantly higher (p<0.05) in PICS-oxy compared to PICS-noxy and woven bags demonstrating the importance of entrapped oxygen on maize carotenoid degradation. Furthermore, differences in carotenoid stability between maize genotypes were observed with OPVI having higher retention than OPVII. After 8 months, carotenoid retention remained dependent on storage bag and genotype with retention being greater in PICS-oxy and PICS-noxy compared to woven bags. However, final levels after 8 months were more similar between storage methods. Overall, oxygen content and genotype were found to be determining factors in the effectiveness of PICS to mitigate carotenoid degradation during post-harvest storage of maize.</p><p>While reducing the rate of carotenoid degradation during postharvest storage of biofortified maize is important, success of biofortified maize is also dependent on consumer adoption of these grains and their performance in traditional food preparation. Assessment of the rheological and functional properties of these two biofortified maize genotypes as a function of post-harvest storage was completed to assess the impact of post-harvest storage in PICS bags on flour functionality and rheological properties for the two biofortified orange maize genotypes and a control white maize genotype. Flour pasting profiles were assessed initially and at 4 and 8 months. After 8 month storage in woven and PICS bag, OPVI and OPVII produced porridges with similar viscosities to their initial viscosities regardless of postharvest storage type. White maize viscosities progressively decreased with storage and were significantly lower (p<0.05) in woven compared to PICS storage. Sequestration of oxygen (PICS-oxy) had modest but significant effects (p<0.05) on key pasting parameters including peak and final viscosities. These results suggest that oxygen sequestration has a critical effect on final flour functionality. DTT treatment partially restored flour pasting profiles suggesting disulfide linkages may modify pasting profiles of flour. There was also an increase in free ferrulic and <i>p</i>-coumaric acids during storage which may have contributed to observed decreases in porridge viscosities. Evidence of this was found through Raman spectroscopy with spectral intensity at both 478cm^-1 and 2911cm^-1 decreasing with storage suggesting the potential for structural changes induced by storage on starch polymer. While storage in PICS bags does not seem to adversely affect flour functionality it may provide some additional economic benefit resulting from requiring proportionally less flour to achieve similar final viscosities as flour from woven bag stored grains. </p><p>Finally, the effect of postharvest storage on bioaccessibility of carotenoids was explored using experimental wet cooked porridges made from ‘fresh’ and stored grains using an established three stage in-<i>vitro</i> digestion model. Relative carotenoid bioaccessibility (% micellarization) was generally higher in less viscous porridge made from grains stored in woven bags compared to porridge from initial or PICS bags stored grains suggesting that higher viscosity might partly explain lower relative bioaccessibility in porridge from grains stored in PICS bags. Absolute carotenoid bioaccessibility from experimental porridge was dependent on carotenoid species and storage system. Extrapolation of relative bioaccessibility (%) to absolute bioaccessibility (µg/g flour) suggests that fresh grains and their corresponding porridges would provide more absolute bioaccessible carotenoids compared to stored grains despite some improvement in relative accessibility. As such, storage losses remain the main factor impacting total available carotenoids and should continue to be an area of focus for future mitigation. With the potential to minimize post-harvest losses, improve carotenoid retention and provide a product with improved cooking performance, PICS bags do appear to offer a viable storage alternative to improve both food and nutrition security in developing countries.</p><p></p>