Carotenoids

Nutritional (ascorbic acid, dehydroascorbic acid and carotenes); antinutritional and toxic components (oxalic acid, nitrate and erucic acid) were determined in sixteen popular species of wild edible plants which are collected for human consumption in southeast Spain. Ascorbic + dehydroascorbic acids contents were very high in several species, especially in Chenopodium album L. (155 mg/100 g). Carotenoid content ranged from 4.2 mg/100 g (Stellaria media Villars) to 15.4 mg/100 g (Amaranthus viridis L.). A range of values was found for oxalic acid from absence to 1100 mg/100 g of plant material. Nitrate contents ranged from 47 mg/100 g (Salicornia europaea L.) to 597 mg/100 g (Amaranthus viridis L.). Low amounts of erucic acid were found in the Cruciferae family (Sisymbrium irio L. 1.73%; Cardaria draba L. 1.23%) and Plantago major L. 3.45%.

Background & aims: Circulating carotenoid levels decrease progressively in patients receiving long-term enteral tube feeding with carotenoid-free formulas. Low dietary intake and low blood levels of carotenoids are associated with a higher risk of morbidity and mortality from chronic diseases. The aim of this study was to examine the effects of a low dose carotenoid mixture (3-mg/1500 kcal) for 3 months on serum carotenoid levels and oxidative stress in patients receiving long-term enteral nutrition as the sole source of nutrition. Methods: This randomized, double blind, controlled study compared patients receiving enteral nutrition with carotenoids (N ¼ 26) and without carotenoids (control group; N ¼ 25). Results: Patients on long-term enteral nutrition had low baseline serum carotenoid levels. Three months of enteral feeding enriched with carotenoids significantly (Po0.01) increased serum carotenoid levels compared with the control group. Oxidative stress as measured by NF-kB levels was decreased at 3 months compared with the control group (Po0.05). No significant changes in MDA levels were observed during the study period in either group. Conclusions: This study demonstrated that enteral nutrition containing small amounts of carotenoids (3-mg/1500 kcal) in patients requiring long-term enteral feeding normalizes serum carotenoid levels to the lower end of the range found in ARTICLE IN PRESS (E.P. Meijer).

Natural compounds are known to modify NO content in tissues; however, the biological activity of polyphenol-rich food often does not correspond to the effects of individual polyphenols on NO synthase activity. The aim of this study was to see how natural compounds luteolin, indole-3-carbinol, and lycopene modify NO production in rat tissues and change the expression of the iNOS gene and protein. Indole-3carbinol produced multiple effects on the NO level; it signifi cantly decreased NO concentration in blood, lungs, and skeletal muscles and increased it in the liver. Indole-3-carbinol enhanced lipopolyssaccharide (LPS)-induced NO production in all rat organs. It decreased iNOS gene expression in the brain cortex of animals that did not receive LPS and up-regulated it in the LPS-treated animals. Lycopene increased the iNOS gene transcription rate in the brain cortex of LPS-treated animals. Luteolin did not modify NO production in any organ of LPS-untreated rats, nor did it affect gene expression in the liver. In the brain it slightly decreased iNOS gene expression. Luteolin decreased NO production in the blood of LPS-treated animals and the number of iNOS-positive cells in these animals. Our results suggest that changes in tissue NO levels caused by natural compounds cannot be predicted from their effect on NOS expression or activity obtained in model systems. This stresses the importance of direct measurements of NO and NOS expression in animal tissues. Rostoka E, et al. EFFECTS OF NATURAL COMPOUNDS ON NO AND iNOS Arh Hig Rada Toksikol 2010;61:275-285

Under chronic stress, carotenoid-based colouration has often been shown to fade. However, the ecological and physiological mechanisms that govern colouration still remain largely unknown. Colour changes may be directly induced by the stressor (for example through reduced carotenoid intake) or due to the activation of the physiological stress response (PSR, e.g. due to increased blood corticosterone concentrations). Here, we tested whether blood corticosterone concentration affected carotenoid-based colouration, and whether a trade-off between colouration and PSR existed. Using the common lizard (Lacerta vivipara), we correlatively and experimentally showed that elevated blood corticosterone levels are associated with increased redness of the lizard's belly. In this study, the effects of corticosterone did not depend on carotenoid ingestion, indicating the absence of a trade-off between colouration and PSR for carotenoids. While carotenoid ingestion increased blood carotenoid concentration, colouration was not modified. This suggests that carotenoid-based colouration of common lizards is not severely limited by dietary carotenoid intake. Together with earlier studies, these findings suggest that the common lizard's carotenoid-based colouration may be a composite trait, consisting of fixed (e.g. genetic) and environmentally elements, the latter reflecting the lizard's PSR. Citation: Fitze PS, Cote J, San-Jose LM, Meylan S, Isaksson C, et al. (2009) Carotenoid-Based Colours Reflect the Stress Response in the Common Lizard. PLoS ONE 4(4): e5111.

In order to assess peripheral levels and activities of a broad spectrum of non-enzymatic and enzymatic antioxidants in elderly subjects with mild cognitive impairment (MCI) and Alzheimer's disease (AD), plasma levels of water-soluble (Vitamin C and uric acid) and of lipophilic (Vitamin A, Vitamin E and carotenoids including lutein, zeaxanthin, ␤-cryptoxanthin, lycopene, ␣and ␤-carotene) antioxidant micronutrients as well as activities of plasma and red blood cell (RBC) superoxide dismutase (SOD) and of plasma glutathione peroxidase (GPx) were measured in 25 patients with MCI, 63 AD patients and 53 controls. Peripheral levels and activities of antioxidants were similarly lower in MCI and AD patients as compared to controls. As MCI may represent a prodromal stage of AD, and oxidative damage appears to occur as one of the earliest pathophysiological events in AD, an increased intake of antioxidants in patients with MCI could be helpful in lowering the risk of conversion to dementia.

Carotenoid-dependent plumage displays are widely assumed to be honest indicators of individual health or quality, which are used as cues during mate choice and/or agonistic signalling. Despite the fact that red, yellow and orange pigmentation of bills is common, and also variable between individuals, comparatively little is known about bill colouration as a condition-dependent trait. Furthermore, many studies of avian colouration are confounded by the lack of objective colour quantification and the use of overly simplistic univariate techniques for analysis of the relationship between the condition-dependent trait and individual quality variables. In this study, we correlated male blackbird bill colour (a likely carotenoid-dependent sexually selected trait) with body/condition variables that reflect male quality. We measured bill colour using photometric techniques, thus ensuring objectivity. The data were analysed using the multivariate statistical techniques of canonical ordination. Analyses based on reflectance spectra of male blackbird bill samples and colour components (i.e. hue, chroma and brightness) derived from the reflectance spectra were very similar. Analysing the entire reflectance spectra of blackbird bill samples with Redundancy Analysis (RDA) allowed examination of individual wavelengths and their specific associations with the body/condition variables. However, hue, chroma and brightness values also provided useful information to explain colour variation, and the two approaches may be complimentary. We did not find any significant associations between male blackbird bill colour and percent incidence of ectoparasites or cloaca size. However, both the colour component and full spectral analyses showed that culmen length explained a significant amount of variation in male blackbird bill colour. Culmen length was positively associated with greater reflectance from the bill samples at longer wavelengths and a higher hue value (i.e. more orange-pigmented bills). Larger males may have larger territories or be better at defending territories during male-male interactions, ensuring access to carotenoid food sources. Future studies should elucidate the relationship between bill colour and behavioural measures such as aggressiveness, territory size, song rate and nest attendance.

Ocular and systemic measurement and imaging of the macular carotenoids lutein and zeaxanthin have been employed extensively as potential biomarkers of AMD risk. In this study, we systematically compare dual wavelength retinal autofluorescence imaging (AFI) of macular pigment with skin resonance Raman spectroscopy (RRS) and serum carotenoid levels in a clinic-based population. Eighty-eight patients were recruited from retina and general ophthalmology practices from a tertiary referral center and excluded only if they did not have all three modalities tested, had a diagnosis of macular telangiectasia (MacTel) or Stargardt disease, or had poor AFI image quality. Skin, macular, and serum carotenoid levels were measured by RRS, AFI, and HPLC, respectively. Skin RRS measurements and serum zeaxanthin concentrations correlated most strongly with AFI macular pigment volume under the curve (MPVUC) measurements up to 9° eccentricity relative to MPVUC or rotationally averaged macular pigment op...

The aim of this study was to assess lycopene intake in daily food rations of women (n = 725) from the Lower Silesia region. The data on the lycopene intake were collected using 24-hour recall and diet history questionnaire. The intake of lycopene was not too low. The average lycopene contents in the daily food rations of 725 women from Lower Silesia region was 7.5 mg. The highest lycopene intake was found in the food daily rations of women with BMI > 30. In examined group of women low knowledge on the importance of the role of lycopene in health was noticed. Educational interventions are required to inform women and to increase lycopene intake among them.

Crocus sativus is a triploid sterile plant characterized by its long red stigmas, which produce and store significant quantities of the apocarotenoids crocetin and crocin, formed from the oxidative cleavage of zeaxanthin. Here, we investigate the accumulation and the molecular mechanisms that regulate the synthesis of these apocarotenoids during stigma development in C. sativus. We cloned the cDNAs for phytoene synthase, lycopene-b-cyclase, and bring hydroxylase from C. sativus. With the transition of yellow undeveloped to red fully developed stigmas, an accumulation of zeaxanthin was observed, accompanying the expression of CsPSY, phytoene desaturase, and CsLYCb, and the massive accumulation of CsBCH and CsZCD transcripts. We analyzed the expression of these two transcripts in relation to zeaxanthin and apocarotenoid accumulation in other Crocus species. We observed that only the relative levels of zeaxanthin in the stigma of each cultivar were correlated with the level of CsBCH transcripts. By contrast, the expression levels of CsZCD were not mirrored by changes in the apocarotenoid content, suggesting that the reaction catalyzed by the CsBCH enzyme could be the limiting step in the formation of saffron apocarotenoids in the stigma tissue. Phylogenetic analysis of the CsBCH intron sequences allowed us to determine the relationships among 19 Crocus species and to identify the closely related diploids of C. sativus. In addition, we examined the levels of the carotenoid and apocarotenoid biosynthetic genes in the triploid C. sativus and its closer relatives to determine whether the quantities of these specific mRNAs were additive or not in C. sativus. Transcript levels in saffron were clearly higher and nonadditive, suggesting that, in the triploid gene, regulatory interactions that produce novel effects on carotenoid biosynthesis genes are involved.

Isolation of carrot fruit (Daucus carrota L., cultivar ''Chanteney") essential oil by supercritical carbon dioxide was investigated from the pretreatment of herbaceous matrix and extraction conditions to the chemical composition of obtained extract and its antimicrobial activity. The qualitative and quantitative analyses of the supercritical extract, as well as of the essential oil obtained by hydrodistillation, were done by GC/FID and GC/MS methods. Antimicrobial properties of both samples were investigated against ten species of microorganisms. Experimental results showed that the particle size had no influence on the extraction process. The highest yield was obtained at 40°C and 10 MPa. The main component of the supercritical extract, as well as of the essential oil was carotol. The supercritical extract was characterized by the presence of heavier molecular weight compounds, while some lighter compounds, e.g. pinenes, were not detected. The supercritical extract and the essential oil were the most effective against Gram-positive bacteria.

A relatively simple but sensitive and reliable method ofquantitating the oxygen-radical absorbing capacity (ORAC) of antioxidants in serum using a few #l is described. In this assay system, ~-phycoerythrin (~-PE) is used as an indicator protein, 2,2'-azobis(2-amidinopropane) dihydrochloride (AAPH) as a peroxyl radical generator, and 6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid (Trolox, a water-soluble vitamin E analogue) as a control standard. Results are expressed as ORAC units, where 10RAC unit equals the net protection produced by l ~M Trolox. The uniqueness of this assay is that total antioxidant capacity of a sample is estimated by taking the oxidation reaction to completion. At this point all of the nonprotein antioxidants (which include a-tocopherol, vitamin C, E-carotene, uric acid, and bilirubin) and most of the albumin in the sample are oxidized by the peroxyl radical. Results are quantified by measuring the protection produced by antioxidants. This solves many problems associated with kinetics or lag-time measurements. A linear correlation of ORAC value with concentration of serum, Trolox, vitamin C, uric acid, and bovine albumin is demonstrated. The coefficient of variation within a run is found to be about 2% and from run to run about 5%. Trolox, a-tocopherol, vitamin C,/3-carotene, uric acid, and bilirubin completely protect B-PE from oxidation, while bovine albumin protects/3-PE only partially. On a molar basis, the relative peroxyl radical absorbance capacity of Trolox, a-tocopherol acid succinate, uric acid, bilirubin, and vitamin C is l : 1 : 0.92 : 0.84 : 0.52. Bovine albumin per unit weight has a lower peroxyi absorbing capacity than these antioxidants. However, the serum protein fraction, containing some lipid-soluble antioxidants, represents the major contributor to the ORAC value found in whole serum. The minimum amount of vitamin C and uric acid which could still be detectable when added to a serum supernatant fraction is 1.5/~g and 0.59 #g, respectively, which account for about 1% of the total ORAC value of the serum supernatant fraction.

Fusarium circinatum is the causal agent of pitch canker disease affecting Pinus spp. and Pseudotsuga menziesii worldwide. Under strict quarantine measures, alternative approaches for disease control are necessary. Phosphite (Phi) salts are known for their fungicidal activity and as plant resistance elicitors; however, its potential is yet to be acknowledged in the Pinus-F. circinatum model. The main aim of this study was to assess whether the application of a Phi-based commercial formulation would delay the progression of the pitch canker on Pinus radiata plants, and on the in vitro fungal growth. In vitro assays were performed using different Phi concentrations (1% and 4%) and a non-treated control (0%), and repeated in vivo using inoculated and non-inoculated plants. Plant physiological parameters and hormonal content were evaluated. Phi was effective at inhibiting in vitro mycelial growth in a dose dependent manner. Regardless of fungal inoculation, Phi application induced positi...

Intracellular antioxidants include low molecular weight scavengers of oxidizing species, and enzymes which degrade superoxide and hydroperoxides. Such antioxidants systems prevent the uncontrolled formation of free radicals and activated oxygen species, or inhibit their ...

Thione analogues of three naturally occurring carotenones (canthaxanthin, echinenone, and rhodoxanthin) were synthesized just over ten years ago, and it was reported that substitution of the oxygen atom by sulphur brings about a large red shift and some broadening in the optical absorption spectrum of the compound. Since the three carotenothiones are scarce, determination of their molar absorption coefficients presents a challenge. A method for relating the molar absorption coefficient of a carotenothione (Car-S) to that of its ketone analogue (Car-O) has been developed, which has revealed that the peak molar absorption coefficient of a Car-S is only about 60% of the corresponding value for Car-O. Using methylene blue as the sensitizer and acetonitrile as the solvent, we have also investigated the quenching (under photostationary conditions) of the 1270 nm phosphorescence emission of singlet oxygen by each of the six carotenoids. The data conform to the Stern-Volmer relation, and show that substitution of a carbonyl oxygen atom by sulphur does not lead to an appreciable change in the value of the quenching constant, which is close to 1.5 · 10 10 M À1 s À1 for all six quenchers.

Inositol-(1,4,5)-trisphosphate (InsP3) is a second messenger in plants that increases in response to many stimuli. The metabolic consequences of this signalling pathway are not known. We reduced the basal level of InsP3 in tomato (Solanum lycopersicum cv. Micro-Tom) by expressing the human type I inositol polyphosphate 5-phosphatase (InsP 5-ptase) gene. Transgenic lines producing InsP 5-ptase protein had between 15% and 30% of the basal InsP3 level of control plants. This increased hydrolysis of InsP3 caused dramatic increases in drought tolerance, vegetative biomass and lycopene and hexose concentrations in the fruits. Transcript profiling of root, leaf and fruit tissues identified a small group of genes, including a cell-wall invertase inhibitor gene, that were differentially regulated in all tissues of the InsP 5-ptase expressing plants. Significant differences were found in the amounts of carbohydrates and organic phosphate in these plants. Plants with increased hydrolysis of InsP3 in the cytosol also showed increased net CO2-fixation and sucrose export into sink tissue and storage of hexoses in the source leaves. The increase in biomass was dependent on the supply of inorganic phosphate in the nutrient medium. Uptake and storage of phosphate was increased in the transgene expressing lines. This suggests that in tomato, increased flux through the inositol phosphate pathway uncoupled phosphate sensing from phosphate metabolism. Altering the second messenger, InsP3, revealed multiple coordinated changes in development and metabolism in tomato that have potential for crop improvement.

The aim of this study was to compare the photoprotective effect of carotenoids in phylogentically related bacteria, which synthesize structurally different pigments. Two organisms were isolated from the same environment. Their 16S rDNA sequences and phenotypic characteristics identified them as members of the family Micrococcaceae. Reverse phase HPLC and absorption spectroscopy revealed that one of them, designated RMB40, synthesized 3 carotenoids with 9 conjugated double bonds, whilst the other, designated RMB42, synthesized a single and more hydrophobic pigment carrying 11 conjugated double bonds. Survival curves were obtained during sunlight exposure for both organisms and for carotenoid deficient mutants derived from them. Increased sunlight sensitivity was found in the carotenoidless mutant derived from RMB42. In contrast, pigment depletion had no appreciable effect on the sunlight response of RMB40. It is concluded that the structure of bacterial carotenoid probably exert an important influence on the effectiveness of these compounds to provide photoprotection in vivo.

Background Individuals consuming diets dense in fruits and vegetables consume an array of phytonutrients as well as recognized nutritional components, including vitamins, minerals, and fiber. There is a growing body of evidence that phytonutrients may play positive roles in health.

Red hues are a challenge in studies on the evolution of bird coloration, as multiple pigments such as carotenoids, pheomelanin, psittacofulvins, porphyrins, turacin, haemoglobin and even exogenous iron-oxides, may confer red colors. Determining the pigment type is paramount and here we investigate the differences in spectrum reflectance for six pigments resulting in red colorations in feathers of different species, with a focus on discriminating among melanins and carotenoids. Pigment chemical identification was obtained from the literature or using High Performance Liquid Chromatography (HPLC) in our laboratory. We have also derived discriminant formulas for identification of the major known types of pigments based on parameters of the reflectance curves obtained with a portable spectrometer. Our results indicate that the reflectance patterns of coloration perceived as red patches widely differ. The distinction between carotenoid-and melanin-based reflectance curves is relatively straightforward: sigmoid versus straight slope. The spectral reflectance curves of feathers containing red psittacofulvins are sigmoid, whereas iron oxide and porphyrin-containing feathers recall pheomelanins in rendering a straight slope. In the case of turacin-based coloration, the spectral shape is unique. For the pigments with enough number of species sampled (i.e., carotenoids, melanins and psittacofulvins) the differences in reflectance shape are important enough to allow separation of carotenoid and melanin derived colorations based on reflectance curves alone.

A comparative study between the antioxidant properties of peel (flavedo and albedo) and juice of some commercially grown citrus fruit (Rutaceae), grapefruit (Citrus paradisi), lemon (Citrus limon), lime (Citrus  aurantiifolia) and sweet orange (Citrus sinensis) was performed. Different in vitro assays were applied to the volatile and polar fractions of peels and to crude and polar fraction of juices: 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging capacity, reducing power and inhibition of lipid peroxidation using b-carotene-linoleate model system in liposomes and thiobarbituric acid reactive substances (TBARS) assay in brain homogenates. Reducing sugars and phenolics were the main antioxidant compounds found in all the extracts. Peels polar fractions revealed the highest contents in phenolics, flavonoids, ascorbic acid, carotenoids and reducing sugars, which certainly contribute to the highest antioxidant potential found in these fractions. Peels volatile fractions were clearly separated using discriminant analysis, which is in agreement with their lowest antioxidant potential.

Xanthophylls are oxygenated carotenoids abun-dant in the human food supply. Lutein, zeaxanthin, and cryptoxanthin are major xanthophyll carotenoids in human plasma. The consumption of these xanthophylls is directly associated with reduction in the risk of ...

Background: Dietary lipids are one of the most effective stimulators of carotenoid absorption, but very limited data exist on the impact of endogenous food sources of lipids to enhance carotenoid absorption. The co-consumption of whole egg with carotenoid-rich foods may increase overall carotenoid absorption via lipid-rich egg yolk. Objective: We designed this study to assess the effects of egg consumption on carotenoid absorption from a carotenoid-rich, raw mixed-vegetable salad. Design: Healthy young men (n = 16) consumed the same salad (all served with 3 g canola oil) with no egg (control), 75 g scrambled whole eggs (1.5 eggs) [low egg (LE)], and 150 g scrambled whole eggs (3 eggs) [high egg (HE)] (a randomized crossover design). Control, LE, and HE meals contained 23 mg, 23.4 mg (0.4 mg from eggs), and 23.8 mg (0.8 mg from eggs) total carotenoids and 3 g, 10.5 g (7.5 g from eggs), and 18 g (15 g from eggs) total lipids, respectively. Blood was collected hourly for 10 h, and the triacylglycerol-rich lipoprotein (TRL) fraction was isolated. Total and individual carotenoid contents, including lutein, zeaxanthin, a-carotene, b-carotene, and lycopene in TRL were analyzed, and composite areas under the curve (AUCs) were calculated. Results: The total mean (6SE) carotenoid AUC 0-10h in TRL was higher for the HE meal than for LE and control meals [125.7 6 19.4 a compared with 44.8 6 9.2 b compared with 14.9 6 5.2 b nmol/ L $ 10 h, respectively (values with superscripts without a common letter differ); P , 0.0001]. The TRL AUC 0-10h of lutein and zeaxanthin increased 4-5-fold (P , 0.001), and the TRL AUC 0-10h of carotenoid not present in eggs, including a-carotene, b-carotene, and lycopene, increased 3-8-fold (P , 0.01) for the HE meal compared with the control meal. Conclusion: These findings support that co-consuming cooked whole eggs is an effective way to enhance carotenoid absorption from other carotenoid-rich foods such as a raw mixed-vegetable salad. This trial was registered at clinicaltrials.gov as NCT01951313.

This work aimed to compare methods for the formation of complexes of bixin and curcumin with β-cyclodextrin (β-CD) and to evaluate the stability of the complexes formed by these methods and their food applications. The stoichiometric relationship between curcumin and β-CD was 1:2 and that between bixin and β-CD was 1:1. Curcumin-β-CD and bixin-β-CD complexes formed by kneading, coprecipitation, and simple mixing were evaluated by differential scanning calorimetry (DSC), thermogravimetry analysis (TGA), or nuclear magnetic resonance (NMR-H). For both curcumin and bixin, the best method of complexation was coprecipitation. Complexation of colorants with β-CD promoted an intensification of color and increased water solubility; however, stabilization in the presence of light occurred only for bixin. Application of curcumin-β-CD in cheese and yogurt and bixin-β-CD in the curd did not alter the initial characteristics of the products, which were sensorialy well accepted. Therefore, the complexation of these natural colorants with β-CD favors their use in low-fat foods, broadening the field of industrial application.

The current study was undertaken to provide solutions to optimize the unsaponifiable antioxidants content of bread. We report a complete description of changes in wheat carotenoids and vitamin E content from grain to bread and highlight the most important processing steps affecting their level in wheat bread. Major carotenoids losses occurred during kneading. A close correlation (r 2 ) 0.97; P ) 0.05) was found between carotenoid pigment losses and lipoxygenase (LOX) activity, both parameters depending on wheat genotype. The use of wheat species exhibiting high carotenoid contents and low LOX activity was shown to preserve significant carotenoid level in the bread. No relation was found between vitamin E losses during doughmaking and LOX activity. In addition, moderate kneading resulted in higher vitamin E retention in comparison with carotenoids (12% and 66% losses, respectively). It is concluded that carotenoids are more susceptible to oxidation by endogenous lipoxygenase than vitamin E during breadmaking. This study showed that bread nutritional quality, in terms of antioxidant content, could be improved by selecting suitable cereal genotypes, if this potential is preserved by milling and baking processes.

The antioxidant capacity of the avian intestinal mucosa is potentially important in protecting the gut wall from the harmful actions of reactive oxygen species originating from the diet, mucosal metabolism and the inflammatory response to enteric microbes. To assess this capacity, we determined the total lipid-soluble and water-soluble antioxidant activities of mucosal extracts, using tissue from different parts of the intestinal tract of the chicken. The lipid-soluble antioxidants, vitamin E and carotenoids, were also measured in the same samples. Total lipid-soluble antioxidant activity was highest in mucosa from the duodenum followed by the jejunum, with much lower activities in the ileum, ceca and colon. Total water-soluble antioxidant activity of the mucosa was at least an order of magnitude greater than the lipid-soluble activity under the assay conditions and did not differ significantly among the different parts of the intestinal tract. High concentrations of vitamin E were present in the mucosa of the duodenum and jejunum, with a trend to lower levels in the ileum and ceca, and significantly less in the colon. Similarly, the mucosa of the duodenum and jejunum contained the highest concentrations of carotenoids, with much lower levels in the ileum and colon. The different isoforms of vitamin E were absorbed from the digesta by the mucosa without any major selectivity. However, the liver was greatly enriched with a-tocopherol over the other isoforms, indicating a high degree of discrimination by this tissue.

T h e use of natural pigments such as lobster carotenoids in fish feed formulations offers advantages over the use of the synthetic alternatives. Microencapsulation of the pigments, with or without the addition of antioxidants to the formulation, may be of benefit in terms of stabilizing pigment colour. In the present study, lobster carotenoids were extracted from lobster shell into petroleum ether and microencapsulated by phase separation and salt coacervation within poly(viny1 alcohol) and poly(viny1 alocohol)/poly( D,L-lactic acid) membranes. Spherical microcapsules, with smooth, thin and resilient membranes were obtained with mean diameters ranging from 50 to 150pm, depending on the membrane material, and source of pigment. The microcapsules were pink-orange in colour, and colour stability was followed spectrophotometrically. Enhanced stability was observed in both membrane materials, in comparison to the non-encapsulated control. Rates of discoloration were determined under a variety of storage conditions, including the absence of light, reduced temperatures and under nitrogen atmosphere. T h e best stability of lobster carotenoids was observed under a nitrogen atmosphere within PVA/PLA membranes, representing an 11-fold enhancement of pigment stability in comparison to the controls. Under ambient conditions, the enhancement in pigment stability was approximately 6-fold. The optimum concentration of PVA during microencapsulation was 3-4%, and the microencapsulated pigments appeared most stable under acidic conditions. T h e rate of discoloration appeared independent of pigment concentration.

Hypotheses of parasite-mediated sexual selection (PMSS) propose that elaborate male ornaments have evolved due to female preferences. Females would benefit from mating with more ornamented males if males' ornamentation signals their health status and ability to provide parasite resistance genes for the offspring. Carotenoid-based plumage coloration of birds has been hypothesised to honestly reflect an individual's health status due to trade-off in allocation of carotenoids between maintenance and signalling functions. The prediction of this hypothesis, namely that individuals with brighter plumage are able to mount stronger immune responses against novel antigens and reveal generally better health state, was tested in captive male greenfinches (Carduelis chloris). Greenfinches with brighter yellow breast feathers showed stronger humoral immune response against novel antigen (SRBC) while no relationship between plumage coloration and an estimate of cellmediated immune responsiveness (PHA response) was detected. Elaborately ornamented individuals had better general health state as indicated by the negative correlations between plumage brightness and heterophil haemoconcentration. Consistent with the concept of PMSS, these results suggest that carotenoid-based plumage coloration in greenfinches honestly signals immunocompetence and health status.

To simultaneously detect Sudan dyes and carotenoids in red pepper and tomato products, high-performance liquid chromatography (HPLC) methods with photodiode-array detection are developed and validated. The methods include the use of end-capped and nonend-capped adsorbents with a gradient elution system starting with water containing methanol. Water content of 9% in the starting mobile phase is found to be necessary to ensure sufficient separation of Sudan dyes and to avoid overlapping or interference with the carotenoids of considerable content. The data of the validation reveal the accuracy and precision of the developed methods. A limit of Sudan dyes detection of 1-5 µg/g in red pepper or tomato sauce could be approached. The methods provide excellent separation of the carotenoids from the unsaponified extracts of red pepper and the tomato products

A central composite design was used to optimize the enzyme-assisted extraction of lycopene from the peel fraction of tomato processing waste. Tomato skins were pretreated by a food-grade enzyme preparation with pectinolytic and cellulolytic activities and then subjected to hexane extraction. The factors investigated included extraction temperature (10-50 • C), pretreatment time (0.5-6.5 h), extraction time (0.5-4.5 h), enzyme solution-to-solid ratio (10-50 dm 3 /kg) and enzyme load (0-0.2 kg/kg). Overall, an 8to 18-fold increase in lycopene recovery was observed compared to the untreated plant material. From a response surface analysis of the data, a second-degree polynomial equation was developed which provided the following optimal extraction conditions: T = 30 • C, extraction time = 3.18 h and enzyme load = 0.16 kg/kg. The obtained results strongly support the idea of using cell-wall degrading enzymes as an effective means for recovering lycopene from tomato waste.

ANTRACT: A carotenoid database for individual and multicomponent foods has been compiled that contains values for the five most common carotenoids (a-carotene, 3carotene,-cryptoxanthin, lycopene, lutein) in 2,458 fruits, vegetables, and multicomponent foods containing fruits and vegetables. The database was used to estimate intakes of specific carotenoids for 19-to 50-year-old women (n = 1,102), using food consumption data obtained from dietary recalls in the US Department of Agriculture Continuing Survey of Food Intake by Individuals, 1986. The major contributors of acarotene were carrots consumed as a single food or as an ingredient in multicomponent foods. Carrots, cantaloupe, and broccoli were the main sources of-carotene. Orange juices and blends, oranges, and tangerines were important contributors of-cryptoxanthin. Tomatoes and tomato products consumed as single foods or as ingredients in multicomponent foods provided most of the dietary lycopene. Contributors of lutein + zeaxanthin included collard, mustard, or turnip greens; spinach; and broccoli. The per capita consumption of total carotenoids (the sum of the five specific carotenoids) among these women was approximately 6 mg/day.

Background: Red-flesh fruit is absent from common sweet orange varieties, but is more preferred by consumers due to its visual attraction and nutritional properties. Our previous researches on a spontaneous red-flesh mutant revealed that the trait is caused by lycopene accumulation and is regulated by both transcriptional and post-transcriptional mechanisms. However, the knowledge on post-transcriptional regulation of lycopene accumulation in fruits is rather limited so far.

Adaptive laboratory evolution is an important tool for the engineering of strains for industrially relevant phenotypes. Traditionally, adaptive laboratory evolution has been implemented to improve robustness of industrial strains under diverse operational conditions; however due to the required coupling between growth and survival, its application for increased production of secondary metabolites generally results in decreased production due to the metabolic burden imposed by, or toxicity of, the produced compound.

Many vertebrates use carotenoid-based signals in social or sexual interactions. Honest signalling via carotenoids implies some limitation of carotenoid-based colour expression among phenotypes in the wild, and at least five limiting proximate mechanisms have been hypothesized. Limitation may arise by carotenoid-availability, genetic constraints, body condition, parasites, or detrimental effects of carotenoids. An understanding of the relative importance of the five mechanisms is relevant in the context of natural and sexual selection acting on signal evolution. In an experimental field study with carotenoid supplementation, simultaneous cross-fostering, manipulation of brood size and ectoparasite load, we investigated the relative importance of these mechanisms for the variation in carotenoid-based coloration of nestling great tits (Parus major). Carotenoid-based plumage coloration was significantly related to genetic origin of nestlings, and was enhanced both in carotenoidsupplemented nestlings, and nestlings raised in reduced broods. We found a tendency for ectoparasite-induced limitation of colour expression and no evidence for detrimental effects of carotenoids on growth pattern, mortality and recruitment of nestlings to the local breeding population. Thus, three of the five proposed mechanisms can generate individual variation in the expression of carotenoid-based plumage coloration in the wild and thus could maintain honesty in a trait potentially used for signalling of individual quality. J . E V O L . B I O L . 1 6 ( 2 0 0 3 ) 9 1 -1 0 0 ª 2 0 0 3 B L A C K W E L L P U B L I S H I N G L T D Variation in carotenoid-based colours 99 J . E V O L . B I O L . 1 6 ( 2 0 0 3 ) 9 1 -1 0 0 ª 2 0 0 3 B L

Phycomyces has been in the laboratories for about 140 years, sometimes following trends and fashions, but often anticipating them. Researchers have been attracted by the sensitive and precise responses of Phycomyces to light and other stimuli, coupled with easy manipulations and good adaptation to laboratory life. It is a simple prototype of the many organisms that use light as a source of information but not as a significant source of energy. Growth, development, genetics, and carotene production have been other subjects of pioneering research. Phycomyces was the second organism, after us, known to require a vitamin. It was one of the first organisms in the research on spontaneous mutants and the second, after Drosophila, in which mutations were induced artificially. It was used to coin the concept and the name of heterokaryosis. Phycomyces heterokaryons offer unique experimental possibilities, for instance in the study of gene function in vivo and the causes of cell death. An overall impression of parsimony and combinatorial gene usage arises from the genetic analysis of the complex functions of this fungus. The main subjects of recent attention have been the various reactions to light, gravitropism, and some aspects of metabolism, particularly the production of carotene. Interest in Phycomyces is slacking because of the repeated failures at transforming it stably with exogenous DNA.

To obtain insight into the genome-wide transcriptional response of heterologous carotenoid production in Saccharomyces cerevisiae, the transcriptome of two different S. cerevisiae strains overexpressing carotenogenic genes from the yeast Xanthophyllomyces dendrorhous grown in carbon-limited chemostat cultures was analysed. The strains exhibited different absolute carotenoid levels as well as different intermediate profiles. These discrepancies were further sustained by the difference of the transcriptional response exhibited by the two strains. Transcriptome analysis of the strain producing high carotenoid levels resulted in specific induction of genes involved in pleiotropic drug resistance (PDR). These genes encode ABC-type and major facilitator transporters which are reported to be involved in secretion of toxic compounds out of cells. β-Carotene was found to be secreted when sunflower oil was added to the medium of S. cerevisiae cells producing high levels of carotenoids, which was not observed when added to X. dendrorhous cells. Deletion of pdr10, one of the induced ABC transporters, decreased the transformation efficiency of a plasmid containing carotenogenic genes. The few transformants that were obtained had decreased growth rates and lower carotenoid production levels compared to a pdr5 deletion and a reference strain transformed with the same genes. Our results suggest that production of high amounts of carotenoids in S. cerevisiae leads to membrane stress, in which Pdr10 might play an important role, and a cellular response to secrete carotenoids out of the cell.

One promising area of current research in chemoprevention is the possibility that micronutrients, including vitamin A analogues, may decrease cancer incidence. The term "vitamin A" refers either to retinol (preformed vitamin A) and its synthetic analogues, or to certain carotenoids (provitamin A), which are converted to retinol in the body, as needed. Retinol and the other retinoids are integrally involved in cell growth and differentiation, which may affect carcinogenesis. Such a role has been supported by a large number of in vitro and animal experiments. Data from studies among humans are sparse, in part because most dietary studies have been conducted in populations in which the vast majority of vitamin A intake is actually beta-carotene and other carotenoids, found in carrots and other vegetables and fruits. Although the carotenoids do not have the hormone-like properties of retinol, they do have a potent antioxidant effect and could thus reduce cancer risk by preventing tissue damage due to oxidation. This possibility is supported by data from a large number of observational epidemiologic studies, most of which have consistently found an inverse relation between consumption of carotene-rich vegetables and cancer risk. However, the only direct way to determine whether carotenoids have a beneficial effect is through large, carefully conducted randomized trials. Several such studies are currently underway and should provide sound evidence on which future medical policy and practice can be based.

Isolation of carrot fruit (Daucus carrota L., cultivar ''Chanteney") essential oil by supercritical carbon dioxide was investigated from the pretreatment of herbaceous matrix and extraction conditions to the chemical composition of obtained extract and its antimicrobial activity. The qualitative and quantitative analyses of the supercritical extract, as well as of the essential oil obtained by hydrodistillation, were done by GC/FID and GC/MS methods. Antimicrobial properties of both samples were investigated against ten species of microorganisms. Experimental results showed that the particle size had no influence on the extraction process. The highest yield was obtained at 40°C and 10 MPa. The main component of the supercritical extract, as well as of the essential oil was carotol. The supercritical extract was characterized by the presence of heavier molecular weight compounds, while some lighter compounds, e.g. pinenes, were not detected. The supercritical extract and the essential oil were the most effective against Gram-positive bacteria.