Algal carotenoids 52; secondary carotenoids of algae 3; carotenoids in a natural bloom of Euglena sanguinea

Quantitative carotenoid analysis of a natural bloom of Euglena sanguinea Ehrenberg revealed the presence of β,β-carotene (1% of total carotenoids), monoesters of adonirubin (3%), diesters of (3S, 3′R)-adonixanthin (13%), diesters of (3S, 3′S)-astaxanthin (75%), 19-monoester of (3R, 3′R, 6R)-loroxanthin (1%), (3R, 3′R)-diatoxanthin (6%), diadinoxanthin (1%) and neoxanthin (traces). The carotenoid content amounted to 0.7% of the dry wt. Methods employed included TLC, HPLC, VIS, MS, CD and H NMR (400 and 500 MHz). The high content of ketocarotenoids is characteristic of secondary carotenoids produced under stressed growth conditions. Previously secondary carotenoids were associated with green algae (Chlorophyceae), but have now been encountered in Euglenophyceae.     

Halocynthiaxanthin and fucoxanthinol isolated from Halocynthia roretzi induce apoptosis in human leukemia, breast and colon cancer cells

The sea squirt Halocynthia roretzi metabolizes fucoxanthin, and subsequently accumulates its derived carotenoids with characteristic structures. In the present study, we isolated halocynthiaxanthin and fucoxanthinol as carotenoids having antiproliferative activity from H. roretzi. Halocynthiaxanthin and fucoxanthinol inhibited the growth of HL-60 human leukemia cells in a dose- and time-dependent manner. Viability of HL-60 treated with 12.5 microM halocynthiaxanthin and fucoxanthinol was decreased by 12.1% and 5.7% of control after 48 h incubation, respectively. Furthermore, halocynthiaxanthin and fucoxanthinol induced apoptosis in HL-60 cells, MCF-7 human breast cancer cells, and Caco-2 human colon cancer cells. When HL-60 cells were incubated with 12.5 microM halocynthiaxanthin and fucoxanthinol for 48 h, relative DNA fragmentations were enhanced to 5- and 7-fold compared to that in control cells, respectively. The activities of apoptosis induction by halocynthiaxanthin and fucoxanthinol were higher than that of fucoxanthin which we have previously reported as a carotenoid possessing the ability to induce apoptosis. Fucoxanthinol exhibited the highest apoptosis-inducing activity among the three carotenoids. Furthermore, the expression levels of apoptosis-suppressing protein Bcl-2 were decreased in HL-60 cells treated with halocynthiaxanthin and fucoxanthinol. These results suggest that halocynthiaxanthin and fucoxanthinol exhibited potential antiproliferative effects via apoptosis induction in several cancer cell lines.     

Carotenoids of Heterosigma akashiwo: A chemosystematic contribution

The presence of C₃₇-norcarotenoids (peridinin and probably pyrrhoxanthin, together 87% of total carotenoids) and the carotenoid pattern in general, including dinoxanthin, diatoxanthin and β,β-carotene, but no fucoxanthin, strongly suggest that H. akashiwo is a dinoflagellate and not a chrysophyte.     

Fucoxanthin metabolites in egg yolks of laying hens

Feeding experiments were conducted with White leghorn laying hens fed a carotenoid depleted control diet (containing some zeaxanthin and lutein) or this diet supplemented with 15% seaweed meal of established carotenoid composition. Egg yolk colour was estimated by use of a Roche Yolk Colour Fan and by detailed quantitative and qualitative carotenoid analysis of individual eggs of three laying hens during 4 weeks. Identification of the carotenoids included HPLC, VIS, MS, ¹H NMR data and partial synthesis. The results confirmed that fucoxanthin, the major carotenoid in seaweed meal, is not transferred to the yolk. However, fucoxanthin gave rise to the metabolites fucoxanthinol, fucoxanthinol 3'-sulphate and paracentrone, that are ascribed to enzymatic modifications occurring in the hens. The difuranoid auroxanthin encountered in the egg yolk was ascribed to violaxanthin and/or its furanoid derivatives present in the seaweed meal. Colour of individual yolks varied considerably. The pigmentation level is discussed.     

Changes in pigment content of the diatom Navicula pelliculosa (Bréb.) Hilse in silicon-starvation synchrony

Summary Exponentially grown cells of the freshwater diatom Navicula pelliculosa (Bréb) Hilse, contained chlorophyll a, chlorophyll c, fucoxanthin, diadinoxanthin, diatoxanthin, neofucoxanthin, -carotene, and an unknown pigment, the absorption spectrum of which is reported. Changes in amounts of chlorophyll a, fucoxanthin and diadinoxanthin were determined during the course of silicon-starvation synchrony carried out in the light or dark. Changes in the rate of chlorophyll a and fucoxanthin syntheses were similar. Synthesis ceased after 5–7 hr of silicon starvation, but recommenced in cultures kept in the light, once silicon was re-introduced. In cultures kept in the dark no significant synthesis was observed after re-introduction of silicon. Diadinoxanthin synthesis continued in the light at all times, although at a lower rate during the silicon-starvation period. In the dark, synthesis of this pigment ceased when cell division stopped, and the amount per unit volume of culture decreased. These results are discussed in relation both to the effect of silicon on the metabolism of the diatom and to the possible function of the carotenoids.Dedicated to Prof. C. B. van Niel on the occasion of his 70th birthday.     

The Composition of Carotenoids in Tissues of the Ascidian <Emphasis Type="Italic">Botryllus schlosseri</Emphasis> (Pallas, 1766) from the Black Sea

The composition of carotenoids was investigated in tissues of the colonial sea squirt Botryllus schlosseri (Pallas, 1766), which inhabits the Crimean coast near the city of Sevastopol. The total carotenoid amount of B. schlosseri was found to be 8.7 ± 4.6 mg/100 g of wet weight. Eleven carotenoids, that is, ß-carotene, 7,8-didehydroastaxanthin, 7,8,7',8',-tetradehydroastaxanthin, pectenolone, 4-ketoalloxanthin, diatoxanthin, diadinoxanthin, fucoxanthin, astaxanthin, alloxanthin, and halocynthiaxanthin, were identified in B. schlosseri with the use of the chromatographic and mass-spectrophotometric methods (UV/Vis, ESITOF/MS, and partially ¹H-NMR). Among these, halocynthiaxanthin (20.8% of the total content of carotenoids), alloxanthin (15.2%), and astaxanthin (12.1%) were found to be the major carotenoids. A comparative analysis of the composition of carotenoids in tissues of B. schlosseri was carried out for different regions of the World Ocean.     

Assessment of grazing by the freshwater copepod Diaptomus minutus using carotenoid pigments: a caution

In order to assess feeding selectivity in freshwater zooplankton, we conducted feeding trials using Diaptomus minutus isolated from two Wisconsin lakes. Copepods were fed an algal assemblage comprised of an equal biomass of a centric diatom, a cryptomonad and a coccal green alga. The total amounts of photopigments were tracked using high-performance liquid chromatography. The removals of carotenoids and a-type phorbins (chlorophyll a and phaeopigments) from feeding suspensions were compared with their presence in the guts of animals, in fecal pellets, and in the final suspension. Diaptomus minutus generally removed either the diatoms primarily or all three algal cells equally. These removals were not reflected in gut extracts of the animals, however, where alloxanthin (marker of cryptomonads) was always present, but where fucoxanthin and diadinoxanthin (markers of diatoms) were never observed. Pigment disappearance was variable for total a-type phorbins, but frequently >90% for carotenoids, particularly for fucoxanthin and diadinoxanthin. Phaeophytin a was the major a-type phorbin detected after gut passage. Our results indicate that evaluations of zooplankton grazing which assume that algal carotenoids remain detectable throughout gut passage must be made with substantial caution and that differences in pigment processing are likely to occur among zooplankton species. Furthermore, grazing experiments designed to evaluate decreases of specific pigments in feeding suspension, rather than their appearance in animals' guts, may prove a more valuable approach to understanding the feeding selectivity of copepods.      

Neoxanthin and fucoxanthinol in Fucus vesiculosus

From a large-scale preparation of the seaweed Focus vesiculosus neoxanthin (Fig. 1A) and fucoxanthinol (Fig. 1B) have been isolated. Mass specta, infrared spectra and chemical reactions showed them to be identical with authentic neoxanthin from Euglena or the p-chloroanil oxidation product of fucoxanthol (Fig. 1C), which was obtained from fucoxanthin (Fig. 1D) after reduction with LiAlH₄. Traces of a pigment having a spectrum similar to that of neochrome (= furanoid neoxanthin) but with different chromatographic properties were also isolated. No pigments of the iso-series were detected of the iso-series were detected in the Fucus extracts. After treatment of fucoxanthin with NaBH₄ seven defined pigments were isolated: neochrome acetate, neochrome, fucoxanthinol, fucoxanthol, and the iso-derivatives isofucoxanthin (Fig. 2A), isofucoxanthinol (Fig. 2B) and isofucoxanthol (Fig. 2C). With sodium ethanolate fucoxanthinol and isofucoxanthinol along with a lot of blue-green degradation products were obtained from fucoxanthin.     

The harmful alga Aureococcus anophagefferens utilizes 19'-butanoyloxyfucoxanthin as well as xanthophyll cycle carotenoids in acclimating to higher light intensities

Aureococcus anophagefferens is a picoplanktonic microalga that is very well adapted to growth at low nutrient and low light levels, causing devastating blooms ("brown tides") in estuarine waters. To study the factors involved in long-term acclimation to different light intensities, cells were acclimated for a number of generations to growth under low light (20μmolphotonsm(-2)s(-1)), medium light (60 or 90μmolphotonsm(-2)s(-1)) and high light (200μmolphotonsm(-2)s(-1)), and were analyzed for their contents of xanthophyll cycle carotenoids (the D pool), fucoxanthin and its derivatives (the F pool), Chls c(2) and c(3), and fucoxanthin Chl a/c polypeptides (FCPs). Higher growth light intensities resulted in increased steady state levels of both diadinoxanthin and diatoxanthin. However, it also resulted in the conversion of a significant fraction of fucoxanthin to 19'-butanoyloxyfucoxanthin without a change in the total F pool. The increase in 19'-butanoyloxyfucoxanthin was paralleled by a decrease in the effective antenna size, determined from the slope of the change in F(0) as a function of increasing light intensity. Transfer of acclimated cultures to a higher light intensity showed that the conversion of fucoxanthin to its derivative was a relatively slow process (time-frame of hours). We suggest the replacement of fucoxanthin with the bulkier 19'-butanoyloxyfucoxanthin results in a decrease in the light-harvesting efficiency of the FCP antenna and is part of the long-term acclimative response to growth at higher light intensities.     

IMPROVED DETECTION AND CHARACTERIZATION OF FUCOXANTHIN‐TYPE CAROTENOIDS: NOVEL PIGMENTS IN EMILIANIA HUXLEYI (PRYMNESIOPHYCEAE)1

A new atmospheric pressure chemical ionization mass spectrometry (APCI‐LC/MS) method improved detection and aided characterization of fucoxanthin related carotenoids, revealing the coccolithophorid Emiliania huxleyi (Lohm.) Hay et Mohler (strain MBA 92, Plymouth) to contain a wider range of acyloxyfucoxanthins than reported previously. The diversity is confirmed as arising from differences in the length of the alkanoic acid substituent esterified at position C‐19′. Acyloxyfucoxanthins with substituents of between four and eight carbons at the C‐19′ position have been detected in a culture of Emiliania huxleyi, where previously only 19′‐butanoyloxyfucoxanthin and 19′‐hexanoyloxyfucoxanthin have been reported in the literature. Novel fucoxanthinol derivatives were also found. The detection of these novel carotenoids in Emiliania huxleyi permits detailed studies of the impact of environmental factors on individual components of the complex pool of fucoxanthin‐type carotenoids in this organism.     

ON PIGMENTS,GROWTH, AND PHOTOSYNTHESIS OF PHAEODACTYLUM TRICORNUTUM12

A maximum growth rate with doubling time of 18 hr at 18 C could be maintained. Continuous cultures at about half maximum growth rate provided cells for study of pigments and photosynthesis. The light intensity curve of photosynthesis had no unusual features and showed light-saturated rates of 30-35 μl O₂/mrn³-hr at 18 C. Pigment analysis showed chlorophylls a and c (a/c ratio = 4), fucoxanthin, β-carotene, and diadinoxanthin. Growth under red light (±660 mμ) altered pigments only by decrease in chlorophyll c to about one-half the content obtained under clear tungsten lamps. The large and anomalous spectral shift in fucoxanthin following organic solvent extraction runs confirmed, but efforts to isolate a native fucoxanthin were unsuccessful. Spectral analysis of acetone extracts and sonicated cell preparations allowed estimate of fractional absorption by each component pigment. The analysis shows that chlorophyll a and fucoxanthin are the principal light absorbing pigments and that absorption by other carotenoids is very small.     

On the absolute configuration of 19′-hexanoyloxyfucoxanthin

The esterifying C₆-acid in 19′-hexanoyloxyfucoxanthin has been identified as n-hexanoic acid by GLC of the methyl ester. Ozonolysis of 19′-n-hexanoyloxyfucoxanthin 3-benzoate provided the n-hexanoyloxy derivative of the allenic ketone produced from fucoxanthin 3-benzoate. NMR and CD correlation of the ozonolysis products and NMR of the native carotenoids provided the basis for assignment of the same absolute configuration of the 19′-n-hexanoyloxy derivative (3S, 5R, 6S, 3′S, 5′R, 6′S) as for fucoxanthin. Biosynthetic implications are considered. CD data for 19′-n-hexanoyloxyfucoxanthin, fucoxanthin and some derivatives thereof are reported. Previously unreported minor carotenoids in Coccolithus huxleyi were diadinoxanthin and 3′-desacetyl 19′-n-hexanoyloxy-fucoxanthin.     

Carotenoids in the sea urchin Paracentrotus lividus: occurrence of 9'-cis-echinenone as the dominant carotenoid in gonad colour determination

Regular sampling of wild Paracentrotus lividus was carried out over a 12-month period to examine seasonal effects on the pigment profile and content of the gonads, especially in comparison to gonad colour. The major pigments detected in the gut wall were breakdown products of fucoxanthin, namely fucoxanthinol and amarouciaxanthin A. Lower levels of other dietary carotenoids (lutein and β-carotene) together with some carotenoids not found in the diet, namely isozeaxanthin and echinenone ( 20% total carotenoid) were also detected in the gut wall. The presence of echinenone in the gut wall demonstrates that this organ acts as a major site of carotenoid metabolism. Echinenone is the dominant carotenoid in the gonads, accounting for approx. 50–60% of the total pigment. Both all-trans and 9′-cis forms of echinenone were detected in both the gut wall and in the gonad, with levels of the 9′-cis form typically 10-fold greater than the all-trans form in the gonad. The detection of large levels of 9′-cis-echinenone in wild sea urchins is unexpected due to the absence of 9- or 9′-cis forms of carotenoids in the natural, algal, diet. Whilst echinenone clearly contributes towards gonad pigmentation, levels of this carotenoid, cannot be directly linked to a qualitative assessment of gonad colour in terms of market acceptability. Indeed, unacceptable gonad colouration can be seen with both very low and high levels of echinenone and total carotenoid. The presence of 9′-cis-echinenone as the major carotenoid contributing to the pigmentation/colour of the gonad is an important observation in terms of developing artificial diets for urchin cultivation.     

Microphytobenthic species composition,pigment concentration,and primary production in sublittoral sediments of the Trondheimsfjord (Norway)1

In spring 2005, monthly sampling was carried out at a sublittoral site near Tautra Island. Microphytobenthic identification, abundance (ABU), and biomass (BIOM), were performed by microscopic analyses. Bacillariophyceae accounted for 67% of the total ABU, and phytoflagellates constituted 30%. The diatom floristic list consisted of 38 genera and 94 species. Intact light‐harvesting pigments chl a, chl c, and fucoxanthin and their derivatives were identified and quantified by HPLC. Photoprotective carotenoids were also observed (only as diadinoxanthin; no diatoxanthin was detected). Average fucoxanthin content was 4.57 ± 0.45 μg fucoxanthin · g sediment dry mass^?1, while the mean chl a concentration was 2.48 ± 0.15 μg · g^?1 dry mass. Both the high fucoxanthin:chl a ratio (considering nondegraded forms) and low amounts of photoprotective carotenoids indicated that the benthic microalgal community was adapted to low light. Microphytobenthic primary production was estimated in situ (MPPs, from 0.15 to 1.28 mg C · m^?2 · h^?1) and in the laboratory (MPPp, from 6.79 to 34.70 mg C · m^?2 · h^?1 under light saturation) as ¹⁴C assimilation; in April it was additionally estimated from O₂‐microelectrode studies (MPP_O2) along with the community respiration. MPP_O2 and the community respiration equaled 22.9 ± 7.0 and 7.4 ± 1.8 mg C · m^?2 · h^?1, respectively. A doubling of BIOM from April to June in parallel with a decreasing photosynthetic activity per unit chl a led us to suggest that the microphytobenthic community was sustained by heterotrophic metabolism during this period.     

GENETIC AND PHYSIOLOGICAL VARIATION IN PIGMENT COMPOSITION OF EMILIANIA HUXLEYI (PRYMNESIOPHYCEAE) AND THE POTENTIAL USE OF ITS PIGMENT RATIOS AS A QUANTITATIVE PHYSIOLOGICAL MARKER

Genetic variation of pigment composition was studied in 16 different strains of Emiliania huxleyi (Lohm.) Hay et Mohler in batch culture. Distinct strain-dependent differences were found in the ratios of fucoxanthin, 19'-hexanoyloxyfucoxanthin, and 19'-butanoyloxyfucoxanthin, hampering the use of these individual pigments as a taxonomic marker at the species level. The molar ratio of total carotenoids to chl a , however, was constant for all strains tested. In addition, the pigment composition of one axenic strain (L) of E. huxleyi at different growth rates in light-, nitrate-, and phosphate-limited continuous cultures was analyzed quantitatively. The pigments fucoxanthin and 19'-hexanoyloxyfucoxanthin correlated closely under all conditions. From steady-state rate calculations, it is hypothesized that 19'-hexanoyloxyfucoxanthin is synthesized from fucoxanthin, with light as a modulating factor. The net rate of synthesis of diatoxanthin depended both on the concentration of diadinoxanthin (its partner in the xanthophyll cycle) and on light, illustrating its photoprotective function in the xanthophyll cycle. In axenic strain L, the ratio of total fucoxanthins to chl a correlated strongly with photon flux density and can potentially be used to assess the physiological status with respect to irradiance in field populations. In multispecific bloom situations, the ratio of diadinoxanthin plus diatoxanthin to total fucoxanthins could be used as an alternative indicator for the light-dependent physiological state of E. huxleyi , provided that no other chromophytes are present. Application of these correlations to mesocosm data from the literature has so far provided no evidence that E. huxleyi blooms form only at inhibiting light levels, as previously suggested.     

Pigments and fatty acids of marine raphidophytes: A chemotaxonomic re-evaluation

The patterns of occurrence of photosynthetic pigments and fatty acids among seven available species (11 strains) of marine raphidophytes were determined and used as chemotaxonomic markers. All currently recognized genera of marine raphidophytes were included for analysis: that is, Chattonella, Fibrocapsa, Heterosigma, Olisthodiscus and Haramonas. The characteristic pigment composition was shown to be chlorophyll a, chlorophylls c₁ and/or c₂, fucoxanthin as the major carot-enoid, β,β-carotene and any or all of zeaxanthin, violaxanthin and an auroxanthin-like pigment as the minor carotenoids. The carotenoid composition of all marine raphidophyte genera investigated was virtually the same, except in Fibrocapsa and Haramonas, which differed due to the occurrence of fucoxanthinol and 19′-butanoyloxyfucoxanthin, respectively. These fucoxanthin derivatives, in addition to fucoxanthin, have potential chemotaxonomic use for differentiating the two species. In all 11 strains, 15 fatty acids (saturated, mono-unsaturated and polyunsaturated) were determined. Significant taxonomic distinctions between genera were reflected by their fatty acid profiles. A rapid key for the differentiation of genera, in addition to morphological features, may be the absence of the 18:4 fatty acid in Olisthodiscus; presence of 18:5 in Heterosigma; the presence of fucoxanthinol in Fibrocapsa and presence of 19′-butanoyloxyfucoxanthin in Haramonas.     

CHLOROPLAST PIGMENT PATTERNS IN DINOFLAGELLATES1

The chlorophylls and carotenoids of 22 species of dinoflagellates were analysed by thin layer chromatography, using 2-dimensional sucrose plates, and 1-dimensional polyethylene plates for chlorophylls c₁ and c₂. Peridinin was the major carotenoid in 19 of the species, while fucoxanthin was the major carotenoid in 3. In the peridinin-containing species, 5 carotenoid fractions, constituting more than 95% of the total carotenoids, were always present. These were peridinin (± neo-peridinin), averaging 64% of the total carotenoid, diadinoxanthin, dinoxanthin, β-carotene and a polar, unidentified pink xanthophyll. Six other carotenoid fractions occurred in minor or trace quantities among the species, but were not identified. Two of these had, a wide distribution; the other 4 were restricted to one or 2 species. The chlorophyll content of the dinoflagellate cultures ranged from 1–141 μg chlorophyll a + c/10⁶ cells, a pattern which was broadly correlated with cell size. In the peridinin-containing species the ratio of chlorophyll a to c on a molar basis was approximately 2 (range 1.60–4.39); in the fucoxanthin-containing species this ratio was approximately 4 (range 2.65–5.73). Both chlorophylls c₁ and c₂ occurred in the fucoxanthin-containing dinoflagellates, and only chlorophyll c₂ (one exception) occurred in the peridinin-containing dinoflagellates. These patterns of chlorophyll c and major carotenoid correspond to patterns previously observed in the Pyrrhophyta and the Chrysophyta, suggesting different phylogenetic origins for the “dinoflagellate” chloroplasts.     

Chlorophyll-Protein Complexes from Euglena gracilis and Mutants Deficient in Chlorophyll b: I. Pigment Composition

The use of n-octyl-beta-d-glucopyranoside along with sodium dodecyl sulfate improves the retention of chlorophyll (Chl) by chlorophyll-protein complexes (CPs) prepared from thylakoids of Euglena gracilis Klebs var bacillaris Cori and yields several additional complexes. Thylakoids from wild-type (WT) cells, solubilized in these detergents and subjected to polyacrylamide gel electrophoresis at 0 degrees C, yield the following CPs, in order of relative molecular weight, containing the pigments shown in parentheses with their respective molar ratios where determined: CP Ia (Chl a, diadinoxanthin and beta-carotene; 100:12:5); CP I (Chl a and beta-carotene; 100:6-12); CPx (Chl and carotenoids); LHCP(2) (light-harvesting CP oligomer) (Chl a, Chl b, diadinoxanthin and neoxanthin; 12:4:3:1); CPy (Chl a, diadinoxanthin and beta-carotene; 100:14:8); CPa (Chl a and beta-carotene; 100:18-25) and LHCP (monomer) (Chl a, Chl b, diadinoxanthin and neoxanthin; 12:6:4:1). The LHCP complexes retain up to 40% of the total Chl and 80% of the Chl b in the thylakoids. CP Ia contains only a trace of Chl b (Chl a/b [mol/mol] = 62). The lower amount of Chl b in Euglena (about 10% of Chl a + b) compared to higher plants (about 30% of Chl a + b) is probably a consequence of the lower Chl b (relative to Chl a) in the LHCPs of Euglena rather than of fewer LHCPs being present. G(1)BU, Gr(1)BSL, and O(4)BSL, mutants of bacillaris low in Chl b (1-2% of Chl a + b), lack the CP Ia, LHCP, and LHCP(2) found in wildtype (WT); G(1) and O(4) also lack CPy. The mutants contain reduced amounts of Chl a (two-thirds of WT in Gr(1) and one-third in G(1) and O(4)) and neoxanthin (20-40% of WT) but retain levels of beta-carotene and diadinoxanthin close to those in cells of WT. The CPs remaining in the mutants have pigment compositions very similar to their counterparts from WT.     

Fucoxanthin regulates adipocytokine mRNA expression in white adipose tissue of diabetic/obese KK-A mice

Fucoxanthin, a marine carotenoid found in edible brown seaweeds, attenuates white adipose tissue (WAT) weight gain and hyperglycemia in diabetic/obese KK-A^y mice, although it does not affect these parameters in lean C57BL/6J mice. In perigonadal and mesenteric WATs of KK-A^y mice fed fucoxanthin, mRNA expression levels of monocyte chemoattractant protein-1 (MCP-1) and tumor necrosis factor-α (TNF-α), which are considered to induce insulin resistance, were markedly reduced compared to control mice. In contrast to KK-A^y mice, fucoxanthin did not alter MCP-1 and TNF-α mRNA expression levels in the WAT of lean C57BL/6J mice. Interleukin-6 (IL-6) and plasminogen activator inhibitor-1 mRNA expression levels in WAT were also decreased by fucoxanthin in KK-A^y mice. In differentiating 3T3-F442A adipocytes, fucoxanthinol, which is a fucoxanthin metabolite found in WAT, attenuated TNF-α-induced MCP-1 and IL-6 mRNA overexpression and protein secretion into the culture medium. In addition, fucoxanthinol decreased TNF-α, inducible nitric oxide synthase (iNOS), and cyclooxygenase-2 (COX-2) mRNA expression in RAW264.7 macrophage-like cells stimulated by palmitic acid. These findings indicate that fucoxanthin regulates mRNA expression of inflammatory adipocytokines involved in insulin resistance, iNOS, and COX-2 in WAT and has specific effects on diabetic/obese KK-A^y mice, but not on lean C57BL/6J mice.     

Carotenoids in petals of perennial Medicago species

The carotenoid content in the petals of fourteen Medicago species was examined, together with eight species studied previously, caratenoids in all the known perennials of the genus are reported. The species can be arranged in relationship groups on the basis of their interfertility. No major carotenoid was species- or groupspecific; a few minor pigments, however, were group- or species-specific. The amount of carotenoids ranged from 7 μg/g dry matter in violet-flowered M. sativa to 2120 μg/g in brownish-yellow M. platycarpos. Xanthophylls constituted 76–99% of the total, with lutein as the major component. β-Carotene, lutein and flavoxanthin were ubiquitous in petals. In M sativa leaves β-carotene, lutein, violaxanthin and neoxanthin constituted 88% of the total. The xanthophylls were esterified in petals but not in leaves.