An extensive body of work has documented the antioxidant role of xanthophylls (lutein and zeaxanthin) in human health and specifically how they provide photoprotection in human vision. More recently, evidence is emerging for the transcriptional regulation of antioxidant response by lutein/lutein cleavage products, similar to the role of β-carotene cleavage products in the modulation of retinoic acid receptors. Supplementation with xanthophylls also provides additional benefits for the prevention of age-related macular degeneration (AMD) and attenuation of Alzheimer's disease symptoms. Mammalian β-carotene oxygenase 2 (BCO2) asymmetrically cleaves xanthophylls as well as β-carotene in vitro. We recently demonstrated that mouse BCO2 (mBCO2) is a functionally palmitoylated enzyme and that it loses palmitoylation when cells are treated with β-carotene. The mouse enzyme is the easiest model to study mammalian BCO2 because it has only one isoform, unlike human BCO2 with several major isoforms with various properties. Here, we used the same acyl-RAC methodology and confocal microscopy to elucidate palmitoylation and localization status of mBCO2 in the presence of xanthophylls. We created large unilamellar vesicle-based nanocarriers for the successful delivery of xanthophylls into cells. We demonstrate here that, upon treatment with low micromolar concentration of lutein (0.15 µM), mBCO2 is depalmitoylated and shows partial nuclear localization (38.00 ± 0.04%), while treatment with zeaxanthin (0.45 µM) and violaxanthin (0.6 µM) induces depalmitoylation and protein translocation from mitochondria to a lesser degree (20.00 ± 0.01% and 35.00 ± 0.02%, respectively). Such a difference in the behavior of mBCO2 toward various xanthophylls and its translocation into the nucleus in the presence of various xanthophylls suggests a possible mechanism for transport of lutein/lutein cleavage products to the nucleus to affect transcriptional regulation.

A simple and accurate reversed phase high-performance liquid chromatography coupled with diode array detector (HPLC-DAD) method for simultaneously determining and quantifying the antioxidants carotenes, xanthophylls, and retinol in human plasma is presented in this paper. Compounds were extracted with hexane, a C30 column, and a mobile phase of methanol, methyl tert-butyl ether, and water were used for the separation of the compounds. A total of 8 carotenoids, 3 Z-β-carotene isomers, and 1 fat-soluble vitamin (retinol) were resolved within 72 min at a flow rate of 0.6 mL/min. Detection was achieved at 450 nm for carotenoids and 330 nm for retinol. To evaluate the effectiveness of themethod, it has been applied to an intervention study conducted on eight volunteers. Results. Limits of detection were between 0.1 μg/mL for lycopene and astaxanthin and 1.3 μg/mL for 15-Z-β-carotene. Recoveries were ranged between 89% and 113% for α-carotene and astaxanthin, respectively. Accuracy was between 90.7% and 112.2% and precision was between 1% and 15% RSD. In human plasma samples compounds studied were identified besides three lycopene isomers, demonstrated to be suitable for application in dietary intervention studies. Conclusions. Due to its accuracy, precision, selectivity, and reproducibility, this method is suitable to dietary habits and/or antioxidants status studies.

Lipase inhibition is one of the directions to control obesity. In vitro assays have confirmed the inhibitory effect of selected xanthophylls, including astaxanthin, fucoxanthinol, fucoxanthin, and neoxanthin. Similarly, an in-silico study also demonstrated the successful inhibition of pancreatic lipase by astaxanthin. Unfortunately, the efficacy of these protocols in the emulsion state typical of lipid digestion remains untested. To address this issue, the current study employed the pH-stat test, which mimics lipid digestion in the gastrointestinal tract, to evaluate native and prepared sea buckthorn and rapeseed oils with varying xanthophyll contents from 0 to 1400 mg/kg oil. Furthermore, a molecular docking of zeaxanthin and violaxanthin (commonly found in plant-based foods), astaxanthin (widely distributed in foods of marine origin) and orlistat (approved as a drug) was performed. The in-silico studies revealed comparable inhibitory potential of all tested xanthophylls (variation from - 8.0 to - 9.3 kcal/mol), surpassing that of orlistat (- 6.5 kcal/mol). Nonetheless, when tested in an emulsified state, the results of pH-stat digestion failed to establish the inhibitory effect of xanthophylls in the digested oils. In fact, lipolysis of native xanthophyll-rich sea buckthorn oil was approximately 22% higher than that of the xanthophyll-low preparation. The key insight derived from this study is that the amphiphilic properties of xanthophylls during the digestion of xanthophyll-rich lipids/meals facilitate emulsion formation, which leads to enhanced fat lipolysis.

Nitzschia laevis is a candidate microorganism for bioactive compounds (fucoxanthin and eicosapentaenoic acid (EPA)) production. In this study, the impacts of glucose-induced trophic transition on biomass, photosynthesis, pigments, and lipid profiles were examined. The specific growth rate was increased under glucose addition, achieved at 0.47 day-1 (0.26 ± 0.01 day-1 for the group without glucose in medium). However, the photosynthetic parameters and pigments including chlorophylls, fucoxanthin, and diatoxanthin were reduced. The net yield of EPA doubled under glucose addition, reaching 20.36 ± 1.22 mg/L in 4 days. In addition, the alteration in detailed lipid molecular species was demonstrated with a focus on EPA-enriched lipids. The effects of 2-deoxyglucose (2DG) indicated that glucose phosphorylation was involved in glucose-induced regulation. These findings provide novel data for guiding the application of this diatom strain in the functional food industries.

Clinical trials report benefits of the xanthophylls lutein and zeaxanthin for skin health. Here a keratinocyte culture was used to investigate the effects of in vitro xanthophyll treatment on gene expression and biochemical pathways.

Fruits and vegetables are pivotal for a healthy diet due partly to their content in bioactive compounds. It is for this reason that we conducted a parallel study to unravel the possible effect on cardiometabolic parameters of the ingestion of anthocyanins, xanthophylls, or both groups of bioactives together in postmenopausal women. Seventy-two postmenopausal women were randomized into an 8-month parallel study: a group consuming 60 mg/day anthocyanins (Group A), a group consuming 6 mg lutein and 2 mg zeaxanthin per day (Group X), and a third group consuming a combination of anthocyanins and xanthophylls in the same amounts (Group A+X). Non-targeted metabolomic analysis was done in plasma samples at baseline and after the 8-month intervention by HPLC-QTOF-MS. Inflammatory, antioxidant, and cardiometabolic parameters were measured at the beginning of the study and after 4 and 8-months intervention. Compared with baseline values, none of the 8-month treatments significantly (p < 0.05) changed systolic or diastolic blood pressure (BP), plasma C-reactive protein, interleukin 6, vascular cell adhesion molecule-1, intercellular adhesion molecule-1, monocyte chemoattractant protein-1 or matrix metalloproteinases 2 and 9. Only plasma glucose levels were significantly decreased by treatment A+X after 8 months, and the plasma metabolomic profile was clearly affected by all three dietary supplementations after 8 months. In parallel, there was an increase, also for the three groups, in the plasma ferric reducing antioxidant power value that did not show any synergistic effect between the two groups of bioactives. Postmenopausal women could benefit from an increase in anthocyanins and xanthophylls intake, through the consumption of fruits and vegetables rich in these two types of compounds. Accordingly, plasma glucose and, above all, the reducing power in plasma, could be improved.

Biofortified maize varieties form an essential part of a nutritious diet; available evidence suggests that different processing methods may affect the final food products. The study aimed to evaluate the effects of processing (roasting) and harvesting time on the bioactive components (lutein, zeaxanthin, β-cryptoxanthin, phytate, tannin and vitamin C) and colour properties (L*, a*, b*), of biofortified orange maize. The orange maize hybrids used for the study were obtained from the International Institute of Tropical Agriculture (IITA) diverse lines with high provitamin A (PVA) content. The results showed that harvesting time and roasting methods significantly (P ≤ 0.001) affected the colour properties. The positive values of ∆b* 30.7, 36.0 and 38.1 at 20 days after pollination (DAP), 27DAP and 34DAP, respectively showed that the intensity of orange colour increased with delay in harvesting time. In unprocessed freshly harvested orange hybrid maize; lutein, zeaxanthin, β-cryptoxanthin, tannin and vitamin C increased with an increase in harvesting time. For roasted hybrid, the mean concentrations of all the bioactive components increased with increases in harvesting time except for tannin and vitamin C that showed a decrease at 20DAP and 27DAP. The results revealed that processing and time of harvest affect the levels of non-provitamin A carotenoids, tannins, phytic acid, Vitamin C and the colour properties of biofortified maize genotypes.

Age-related macular degeneration (AMD) is nowadays considered among the retinal diseases whose clinical management lacks established treatment approaches, mainly for its atrophic (dry) form. In this respect, the use of dietary patterns enriched in omega-3 and antioxidant xanthophylls has emerged as a promising approach to counteract dry AMD progression although the prophylactic potential of omega-3 of fish origin has been discussed. Whether enriched availability of omega-3 and xanthophylls may increase the effectiveness of diet supplementation in preventing dry AMD remains to be fully established. The present study aims at comparing the efficacy of an existing orally administered formulation based on lutein and fish oil, as a source of omega-3, with a novel formulation providing the combination of lutein and astaxanthin with Calanus oil (COil), which contains omega-3 together with their precursors policosanols. Using a mouse model of dry AMD based on subretinal injection of polyethylene glycol (PEG)-400, we assessed the comparative efficacy of both formulations on PEG-induced major hallmarks including oxidative stress, inflammation, glial reactivity and outer retinal thickness. Dietary supplementation with both mixtures has been found to exert a significant antioxidant and anti-inflammatory activity as reflected by the overall amelioration of the PEG-induced pathological hallmarks. Noteworthy, the formulation based on COil appeared to be more protective than the one based on fish oil, presumably because of the higher bioavailability of omega-3 in COil. These results support the use of dietary supplements combining omega-3 and xanthophylls in the prevention and treatment of AMD and suggest that the source of omega-3 might contribute to treatment efficacy.

Reduction of nitrogen application in crop production is desirable for ecological and health-related reasons. Interestingly, nitrogen deficiency can lead to enhanced concentrations of polyphenols in plants. The reason for this is still under discussion. The plants' response to low nitrogen concentration can interact with other factors, for example radiation intensity. We cultivated red and green leaf lettuce hydroponically in a Mediterranean greenhouse, supplying three different levels of nitrogen (12 mM, 3 mM, 0.75 mM), either in full or reduced (-50%) radiation intensity. In both red and green lettuce, we found clear effects of the nitrogen treatments on growth characteristics, phenolic and photosynthetic compounds, nitrogen, nitrate and carbon concentration of the plants. Interestingly, the concentrations of all main flavonoid glycosides, caffeic acid derivatives, and sucrose increased with decreasing nitrogen concentration, whereas those of chlorophylls, β-carotene, neoxanthin, lactucaxanthin, all trans- and cis-violaxanthin decreased. The constitutive concentrations of polyphenols were lower in the green cultivar, but their relative increase was more pronounced than in the red cultivar. The constitutive concentrations of chlorophylls, β-carotene, neoxanthin, all trans- and cis-violaxanthin were similar in red and green lettuce and with decreasing nitrogen concentration they declined to a similar extent in both cultivars. We only detected little influence of the radiation treatments, e.g. on anthocyanin concentration, and hardly any interaction between radiation and nitrogen concentration. Our results imply a greater physiological plasticity of green compared to the red lettuce regarding its phenolic compounds. They support the photoprotection theory regarding anthocyanins as well as the theory that the deamination activity of phenylalanine ammonia-lyase drives phenylpropanoid synthesis.

The xanthophyll cycle is a regulatory mechanism operating in the photosynthetic apparatus of plants. It consists of the conversion of the xanthophyll pigment violaxanthin to zeaxanthin, and vice versa, in response to light intensity. According to the current understanding, one of the modes of regulatory activity of the cycle is associated with the influence on a molecular organization of pigment-protein complexes. In the present work, we analyzed the effect of violaxanthin and zeaxanthin on the molecular organization of the LHCII complex, in the environment of membranes formed with chloroplast lipids. Nanoscale imaging based on atomic force microscopy (AFM) showed that the presence of exogenous xanthophylls promotes the formation of the protein supramolecular structures. Nanoscale infrared (IR) absorption analysis based on AFM-IR nanospectroscopy suggests that zeaxanthin promotes the formation of LHCII supramolecular structures by forming inter-molecular β-structures. Meanwhile, the molecules of violaxanthin act as "molecular spacers" preventing self-aggregation of the protein, potentially leading to uncontrolled dissipation of excitation energy in the complex. This latter mechanism was demonstrated with the application of fluorescence lifetime imaging microscopy. The intensity-averaged chlorophyll a fluorescence lifetime determined in the LHCII samples without exogenous xanthophylls at the level of 0.72 ns was longer in the samples containing exogenous violaxanthin (2.14 ns), but shorter under the presence of zeaxanthin (0.49 ns) thus suggesting a role of this xanthophyll in promotion of the formation of structures characterized by effective excitation quenching. This mechanism can be considered as a representation of the overall photoprotective activity of the xanthophyll cycle.

Xanthophylls are oxygenated carotenoids playing an essential role as structural components of the photosynthetic apparatus. Xanthophylls contribute to the assembly and stability of light-harvesting complex, to light absorbance and to photoprotection. The first step in xanthophyll biosynthesis from α- and β-carotene is the hydroxylation of ε- and β-rings, performed by both non-heme iron oxygenases (CHY1, CHY2) and P450 cytochromes (LUT1/CYP97C1, LUT5/CYP97A3). The Arabidopsis triple chy1chy2lut5 mutant is almost completely depleted in β-xanthophylls.

Petunia (Petunia hybrida) is an important ornamental plant with a wide range of corolla colors. Although pale-yellow-flowered cultivars, with a low amount of carotenoids in their corollas, are now available, no deep-yellow-flowered cultivars exist. To find why petunia cannot accumulate enough carotenoids to have deep-yellow flowers, we compared carotenoid profiles and expression of carotenoid metabolic genes between pale-yellow-flowered petunia and deep-yellow-flowered calibrachoa (Calibrachoa hybrida), a close relative. The carotenoid contents and the ratios of esterified xanthophylls to total xanthophylls in petunia corollas were significantly lower than those in calibrachoa, despite similar carotenoid components. A lower esterification rate of trans-xanthophylls than of cis-xanthophylls in petunia suggests that petunia xanthophyll esterase (XES) has low substrate specificity for trans-xanthophylls, which are more abundant than cis-xanthophylls in petunia corolla. The expression of genes encoding key enzymes of carotenoid biosynthesis was lower and that of a carotenoid catabolic gene was higher in petunia. XES expression was significantly lower in petunia. The results suggest that low biosynthetic activity, high cleavage activity, and low esterification activity cause low carotenoid accumulation in petunia corollas.

Xanthophylls are oxygenated carotenoids and fulfill critical roles in plant growth and development. In plants, two different types of carotene hydroxylases, non-heme di-iron and heme-containing cytochrome P450, were reported to be involved in the biosynthesis of xanthophyll. Citrus fruits accumulate a high amount of xanthophylls, especially β,β-xanthophylls. To date, however, the roles of carotene hydroxylases in regulating xanthophyll content and composition have not been elucidated.

Xanthophylls, oxygenated derivatives of carotenes, play critical roles in photosynthetic apparatus of cyanobacteria, algae, and higher plants. Although the xanthophylls biosynthetic pathway of algae is largely unknown, it is of particular interest because they have a very complicated evolutionary history. Carotenoid hydroxylase (CHY) is an important protein that plays essential roles in xanthophylls biosynthesis. With the availability of 18 sequenced algal genomes, we performed a comprehensive comparative analysis of chy genes and explored their distribution, structure, evolution, origins, and expression.

The majority of carotenoids in petals are xanthophylls and most of these xanthophylls are esterified with fatty acids. Although petunia (Petunia x hybrida) is an important ornamental plant, it cannot accumulate enough carotenoids to have deep-yellow flowers. Our previous study suggested that low esterification activity causes low carotenoid accumulation in petunia corollas. Here, we introduced xanthophyll esterase (XES) from the petals of Ipomoea obscura, tomato (Solanum lycopersicum), and marigold (Tagetes erecta) into a pale-yellow-flowered cultivar of petunia to see whether these affect carotenoid accumulation in petunia corollas. Carotenoid contents and the proportions of esterified xanthophylls were elevated in the corollas of XES-overexpressing (XES-OX) transformants. Expression analysis showed that the transcript levels of endogenous carotenoid biosynthetic genes, which included geranylgeranyl diphosphate synthase 2, ζ-carotene desaturase, and lycopene β-ring cyclase in corolla tubes were upregulated in XES-OX plants. In addition, we discovered a difference in the composition of esterified xanthophylls among XES-OX plants, which may be caused by differences in the substrate specificity of their respective XESs. We conclude that esterification is an important process for carotenoid accumulation and XES is a useful tool for the quantitative and qualitative control of carotenoid accumulation in petals.

Carotenoids are natural products regulated by the food sector, currently used as feed dyes and as antioxidants in dietary supplements and composing functional foods for human consumption. Of the nearly one thousand carotenoids described to date, only retinoids, derived from beta carotene, have the status of a drug and are regulated by the pharmaceutical sector. In this review, we address a novel field: the transformation of xanthophylls, particularly the highly marketed astaxanthin and the practically unknown bacterioruberin, in therapeutic agents by altering their pharmacokinetics, biodistribution, and pharmacodynamics through their formulation as nanomedicines. The antioxidant activity of xanthophylls is mediated by routes different from those of the classical oral anti-inflammatory drugs such as corticosteroids and non-steroidal anti-inflammatory drugs (NSAIDs): remarkably, xanthophylls lack therapeutic activity but also lack toxicity. Formulated as nanomedicines, xanthophylls gain therapeutic activity by mechanisms other than increased bioavailability. Loaded into ad hoc tailored nanoparticles to protect their structure throughout storage and during gastrointestinal transit or skin penetration, xanthophylls can be targeted and delivered to selected inflamed cell groups, achieving a massive intracellular concentration after endocytosis of small doses of formulation. Most first reports showing the activities of oral and topical anti-inflammatory xanthophyll-based nanomedicines against chronic diseases such as inflammatory bowel disease, psoriasis, atopic dermatitis, and dry eye disease emerged between 2020 and 2023. Here we discuss in detail their preclinical performance, mostly targeted vesicular and polymeric nanoparticles, on cellular models and in vivo. The results, although preliminary, are auspicious enough to speculate upon their potential use for oral or topical administration in the treatment of chronic inflammatory diseases.

Diets high in fruits and vegetables are generally believed protective against several chronic diseases. One suggested mechanism is a reduction in oxidative stress. The carotenoids, nutrients found in colored fruits and vegetables, possess antioxidant properties in vitro, but their role in humans is less well documented. The aim of this cross-sectional study was to explore the relationships between the most abundant plasma carotenoids (alpha-carotene, beta-carotene, lycopene, lutein, zeaxanthin and beta-cryptoxanthin), as well as grouped carotenoids (total xanthophylls, carotenes and carotenoids), and urinary excretion of the F(2)-isoprostanes (F(2)-IsoPs), stable and specific biomarkers of oxidative damage to lipids. Two F(2)-IsoP measures were utilized: total F(2)-IsoPs and 8-iso-PGF(2alpha). The study population (N = 52) was drawn from a study among patients curatively treated for early-stage head and neck cancer. Unadjusted linear regression analyses revealed significant inverse associations between plasma lutein, total xanthophylls and both F(2)-IsoP measures at baseline. After control for potential confounders, all individual and grouped xanthophylls remained inversely associated with the F(2)-IsoP measures, but none of these associations achieved significance. The carotenes were not inversely associated with total F(2)-IsoPs or 8-iso-PGF(2a) concentrations. The finding of consistent inverse associations between individual and grouped xanthophylls, but not individual and grouped carotenes, and F(2)-IsoPs is intriguing and warrants further investigation.

Red alga species belonging to the Porphyra and Pyropia genera (commonly known as Nori), which are widely consumed and commercialized due to their high nutritional value. These species have a carotenoid profile dominated by xanthophylls, mostly lutein and zeaxanthin, which have relevant benefits for human health. The effects of different abiotic factors on xanthophyll synthesis in these species have been scarcely studied, despite their health benefits. The objectives of this study were (i) to identify the abiotic factors that enhance the synthesis of xanthophylls in Porphyra/Pyropia species by conducting a systematic review and meta-analysis of the xanthophyll content found in the literature, and (ii) to recommend a culture method that would allow a significant accumulation of these compounds in the biomass of these species. The results show that salinity significantly affected the content of total carotenoids and led to higher values under hypersaline conditions (70,247.91 µg/g dm at 55 psu). For lutein and zeaxanthin, the wavelength treatment caused significant differences between the basal and maximum content (4.16-23.47 µg/g dm). Additionally, in Pyropia spp., the total carotenoids were considerably higher than in Porphyra spp.; however, the lutein and zeaxanthin contents were lower. We discuss the specific conditions for each treatment and the relation to the ecological distribution of these species.

Absorbed energy in excess of that used by photosynthesis induces photoinhibition, which is common in water deficit conditions, resulting in reductions in stomatal conductance. In grapevines, controlled water deficit is a common field practice, but little is known about the impact of a given water shortage on the energy transduction processes at the leaf level in relation to contrasting stomatal sensitivities to drought. Here, we assessed the effect of a nearly similar water deficit condition on four grapevine varieties: Cabernet Sauvignon (CS) and Sauvignon Blanc (SB), which are stomatal sensitive, and Chardonnay (CH) and Carménère (CM), which are less stomatal sensitive, grown in 20 L pots outdoors. Plants were maintained to nearly 94% of field capacity (WW) and 83% field capacity (WD). We have assessed plant water status, photosynthesis (AN), photorespiration, AN vs. PAR, ACi curves, photochemical (qP) and non-photochemical (qN) fluorescence quenching vs. PAR, the photoprotective effectiveness of NPQ (qPd) and light interception by leaves. Photorespiration is important under WD, but to a different extent between varieties. This is related to stomatal sensitivity, maintaining a safe proportion of PSII reaction centres in an open state. Additionally, the capacity for carboxylation is affected by WD, but to a greater extent in more sensitive varieties. As for qN, in WD it saturates at 750 μmol PAR m-2s-1, irrespective of the variety, which coincides with PAR, from which qN photoprotective effectiveness declines, and qP is reduced to risky thresholds. Additionally, that same PAR intensity is intercepted by WD leaves from highly stomatal-sensitive varieties, likely due to a modification of the leaf angle in those plants. Pigments associated with qN, as well as chlorophylls, do not seem to be a relevant physiological target for acclimation.

Simultaneous genome editing of the two homeologous LCYe and ZEP genes of Nicotiana benthamiana results in plants in which all xanthophylls are replaced by zeaxanthin. Plant carotenoids act both as photoreceptors and photoprotectants in photosynthesis and as precursors of apocarotenoids, which include signaling molecules such as abscisic acid (ABA). As dietary components, the xanthophylls lutein and zeaxanthin have photoprotective functions in the human macula. We developed transient and stable combinatorial genome editing methods, followed by direct LC-MS screening for zeaxanthin accumulation, for the simultaneous genome editing of the two homeologous Lycopene Epsilon Cyclase (LCYe) and the two Zeaxanthin Epoxidase (ZEP) genes present in the allopolyploid Nicotiana benthamiana genome. Editing of the four genes resulted in plants in which all leaf xanthophylls were substituted by zeaxanthin, but with different ABA levels and growth habits, depending on the severity of the ZEP1 mutation. In high-zeaxanthin lines, the abundance of the major photosystem II antenna LHCII was reduced with respect to wild-type plants and the LHCII trimeric state became unstable upon thylakoid solubilization. Consistent with the depletion in LHCII, edited plants underwent a compensatory increase in PSII/PSI ratios and a loss of the large-size PSII supercomplexes, while the level of PSI-LHCI supercomplex was unaffected. Reduced activity of the photoprotective mechanism NPQ was shown in high-zeaxanthin plants, while PSII photoinhibition was similar for all genotypes upon exposure to excess light, consistent with the antioxidant and photoprotective role of zeaxanthin in vivo.