Metabolism of carotenoids in sea-urchin Pseudocentrotus depressus

• 1.1. Feeding experiments with β,β-carotene, canthaxanthin and astaxanthin on the sea urchin Pseudocentrotus depressus were investigated.
• 2.2. In the case of β,β-carotene group, β-carotene was accumulated, β-isocryptoxanthin appeared and β-echinenone increased 6.8 times as much as the control group. On the other hand, in canthaxanthin and astaxanthin groups, canthaxanthin and astaxanthin increased significantly, respectively. The metabolic products of these carotenoids could not be found.
• 3.3. It was concluded that β,β-carotene was bioconverted to β-echinenone via β-isocryptoxanthin in P. depressus and could not be oxidatively metabolized beyond β-echinenone.

     

Comparative biochemical studies of carotenoids in sea-urchins—II. The more primitive sea-urchins belonging to the orders Cidaroida,Echinothurioida, Diadematoida and Arbacioida

• 1.1. The carotenoids of seven species of more primitive sea-urchins, [orders Cidaroida (I), Echinothurioida (II), Diadematoida (III), and Arbacioida (IV)] were investigated from the comparative biochemical point of view.
• 2.2. β,β-carotene and β-echinenone have been isolated as major carotenoids in (I) and (III, IV), respectively. In (II), β,β-carotene, β-echinenone, canthaxanthin and (3S,3′S)-astaxanthin were foundto be predominant carotenoids.
• 3.3. The carotenoid patterns of (I) which is the most primitive sea-urchin from the phylogenetic point of view, and of (II) which is direct developers with non-feeding larvae, were quite different from those of the other sea-urchins showing typical development with feeding larvae.

     

Characterization of a blue astaxanthin protein from the carapace of the crayfish Astacus leptodactylus

• 1.1. A blue carotenoprotein (λ_max = 634 nm) containing astaxanthin as prosthetic group, was extracted and purified from the carapace of the crayfish Astacus leptodactylus.
• 2.2. The blue carotenoprotein contained (3S,3′S)-astaxanthin, (3R,3′S, meso)-astaxanthin and (3R,3′R)-astaxanthin in relative ratio 38:41:21.
• 3.3. The blue carotenoprotein had an approximate mol. wt of 440,000 (gel filtration) and 437,000 (gradient gel electrophoresis).
• 4.4. Sodium dodecyl sulphate polyacrylamide gel electrophoresis indicated the presence of two polypeptides of 19,600 and 18,600 daltons, with different mobility in polyacrylamide gel electrophoresis in the presence of 6 M urea.
• 5.5. At low ionic strength and in the presence of denaturing agents such as SDS, urea, extreme pH and heat, the blue complex showed a greater stability than most of the carotenoproteins studied to date.

     

Carotenoids in fish IV. Salmonidae and Thymallidae from Polish waters

The author investigated the presence of various carotenoids in the Salmonidae and Thymallidae family by means of columnar and thin-layer chromatography. The investigations revealed the presence of the following carotenoids:

Abstract

- in the muscles of Salmo salar: astaxanthin (pure and ester), canthaxanthin, lutein and zeaxanthin.

- in the eggs of Salmo trutta m. trutta: β-carotene, iso- and zeaxanthin, lutein, taraxanthin and astaxanthin.

- in the eggs of Salmo trutta m. fario: β-carotene, canthaxanthin, 4-keto-4-hydroxy-β-carotene, astaxanthin (pure and ester), lutein, taraxanthin and astacene.

- in the eggs of Salmo gairdneri: β-carotene, γ-carotene (?), canthacanthin, isozeaxanthin, lutein and astaxanthin, and in the sperm Salmo gairdneri: β-carotene, γ-carotene (?), 4-keto-4-hydroxy-β-carotene, canthaxanthin, lutein and astaxanthin.

- in the eggs of Salvelinus fontinalis: ester astaxanthin, canthaxanthin, isozeaxanthin, lutein and astacene.

- in the eggs of Hucho hucho: β-carotene, tunaxanthin, lutein, taraxanthin and astaxanthin.

- in the eggs of Coregonus albula: β-carotene, 4-keto-4-hydroxy-β-carotene, ester astaxanthin, zeaxanthin, taraxanthin and astacene.

- in Coregonus lavaretus: a) in eggs: β-carotene, ester astaxanthin, canthaxanthin, iso- and zeaxanthin, lutein, taraxanthin and astacene b) in the sperm: canthaxanthin, 4-hydroxy-4-keto-β-carotene, isozeaxanthin and astaxanthin, and other organs: 4-hydroxy-α-carotene, canthaxanthin, tunaxanthin, monoepoxy lutein, lutein, iso- and zeaxanthin and astaxanthin.

- in the eggs of Coregonus peled: β-carotene, 4-keto-4-hydroxy-β-carotene, lutein, zeaxanthin, taraxanthin and astacene.

- in the eggs of Thymallus thymallus: β-carotene, tunaxanthin, lutein and astaxanthin.

     

Bioconversion pathway of astaxanthin into retinol2 in mature rainbow trout (Salmo Gairdneri Rich.)

• 1.1. Carotenoid and retinoid forms were analysed by HPLC in various tissues of mature rainbow trout fed for 11 months (7–9°C) with astaxanthin (50 and 100 mg/kg diet) or canthaxanthin (100 mg/kg diet).
• 2.2. Decreasing concentrations of canthaxanthin, echinenone and β-carotene, but no retinol₁, were found in the liver and skin of canthaxanthin-fed fish.
• 3.3. Higher retinol₂ concentrations were found in ovaries and testes of astaxanthin-fed fish compared to canthaxanthin and control groups.
• 4.4. A new metabolic pathway for direct conversion of astaxanthin into retinol₂ in gonads is proposed.

     

Characteristics of carotenoid features in muscle and ovary from anadromous and river resident types of Alaskan dolly varden charr (Salvelinus malma malma)

• 1.1. The carotenoids in the muscles and eggs from two types of natural Alaskan dolly varden charr, anadromous and river resident types, were examined.
• 2.2. In the muscle from the anadromous charr, astaxanthin was the major component comprising more than 70% of the total, followed by idoxanthin and 4-keto-zeaxanthin.
• 3.3. The egg carotenoid features in the river resident charr were more complicated compared with those in the anadromous fish. A considerable proportion of unidentified carotenoids was found in the eggs from the river resident charr.
• 4.4. Idoxanthin was the main component along with considerable propotions of β-carotene tetrol and astaxanthin in the eggs from the anadromous charr, whereas zeaxanthin and lutein were detected besides idoxanthin and β-carotene tetrol in the eggs from the river resident fish.
• 5.5. The prey was considered to be responsible for the difference in the carotenoid features of the eggs from the anadromous and river resident charr.

     

Comparative studies of carotenoids in the fauna of the Gullmar Fjord (Bohuslan,Sweden) III. Echinodermata

The author investigated the presence of various carotenoids in the Echinodermata from Gullmar Fjord (Bohuslan, Sweden) by means of columnar and thin-layer chromatography. The investigations revealed the presence of the following:

- inHenricia sanguinolenta:β-carotene, echinenone, canthaxanthin, guraxanthin, lutein-5, 6-epoxide and astaxanthin.

- inAmphiura filiformis: canthaxanthin, cryptoxanthin, lutein, lutein-5, 6-epoxide, isozeaxanthin, zeaxanthin, astaxanthin and 4-hydroxy-4-keto-β-carotene.

- inAmphipholis squamata:β-carotene, cryptoxanthin, lutein, lutein-5, 6-epoxide, astaxanthin, astaxanthin ester, asterin-acid and rubixanthin derivative.

- inOphiopholis aculeata: canthaxanthin, cryptoxanthin, isozeaxanthin, astaxanthin, astaxanthin ester, asterinacid, 4-hydroxy-4-keto-β-carotene, hydroxy rubixanthin and gazaniaxanthin-like substances.

- inOphiothrix fragilis: canthaxanthin, lutein-5, 6-epoxide, isozeaxanthin, astaxanthin, astaxanthin ester, 4-hydroxy-4-keto-β-carotene, and hydroxy rubixanthin.

- inAntedon petatus:canthaxanthin, guaraxanthin, isozeaxan-thin, zeaxanthin, astaxanthin, astaxanthin ester and 4-keto-4-ethoxy-β-carotene.

- inEchinocardium cordatum:β-carotene,γ-carotene, canthaxanthin, lutein, isozeaxanthin, zeaxanthin, astaxanthin and astaxanthin ester.

- inSpatangus purpureus: isozeaxanthin, astaxanthin, astaxanthin ester and 4-hydroxy-4-keto-β-carotene.

     

Comparative biochemical studies of carotenoids in fish—XXVII. Carotenoidsin the eggs of three species of cyprinidae

• 1.1. The carotenoids in the eggs of three species of freshwater fish, carp Cyprinus carpio, silver crucian carp Carassius gibelio langsdorfi and oily gudgeon Sarocheilichthys variegatus belonging to Cyprinidae were investigated from the comparative biochemical point of view.
• 2.2. Lutein A (3), (3R,3′R)-zeaxanthin (4), diatoxanthin (5) and alloxanthin (6) were found to be dominant and their corresponding 4-keto and/or 4′-keto derivatives, i.e. fritschiellaxanthin (10), (3S,3′R)-4-ketozeaxanthin (11), (3S,3′S)-astaxanthin (12), (3S,3′S)-7,8-didehydroastaxanthin (13) and (3S,3′S)-7,8,7′,8′-tetradehydroastaxanthin (14) were also found in all species examined.

     

Carotenoids in fish. XXVI. Pungitius pungitius (L.) and Gasterosteus aculeatus L. (Gasterosteidae)

The author investigated the presence of various carotenoids in the different parts of the body of Pungitius pungitius (L.) and Gasterosteus aculeatus L. by means of columnar and thin-layer chromatography. The investigations revealed the presence of the following carotenoids:

in Pungitius pungitius. α-carotene, β-carotene, β-cryptoxanthin, mutatochrome, zeaxanthin and astaxanthin;

in Gasterosteus aculeatus: β-carotene, β-cryptoxanthin, β-carotene epoxide, neothxanthin, canthaxanthin, mutatochrome, lutein, phoenicoxanthin, zeaxanthin, taraxanthin, tunaxanthin, astaxanthin, astaxanthin ester and α-doradexanthin. The total carotenoid content ranged from 2.229 to 138.504 µg/g wet weight.

     

Distribution of canthaxanthin in immature rainbow trout Oncorhynchus mykiss serum

• 1.1. The present study was undertaken in order to define the distribution of canthaxanthin between the lipoprotein fractions in serum of immature rainbow trout fed a diet supplemented with synthetic canthaxanthin (80 mg/kg).
• 2.2. Lipoproteins were separated by density-gradient ultracentrifugation.
• 3.3. Canthaxanthin was found in all lipoprotein fractions, in different amounts according to the density of the lipoprotein fraction: VLDL, 13.9%; LDL, 15.2% or LDL, 29.1% since the density of the first fraction was 1.006 g/ml; HDL, 60.4% and VHDL, 10.5%.

     

Precence of β-Carotene and Xantophylls in two species of the genus Niphargus (Crustacea: Amphipoda)

The presence of carotenoids in the Niphargus tatrensis Wrzesniowski and Niphargus aquilex schellenbergi Karaban has been investigated. In extracts separated by means of column and thin-layer chromatography, the following carotenoids were identified:

1. in Niphargus tatrensis: β-carotene, astaxanthin ester, rubixanthin, celaxanthin and astaxanthin.
2. in Niphargus aquilex schellenbergi: cantha-xanthin, astaxanthin ester, isozeaxanthin (only from Jeker), 4-keto-4′-hydroxy-β-carotene (only from Terzietebronbos), rubixanthin, celaxanthin and astaxanthin.

It was not possible to identifity the sexth fractions.     

Comparative study on the fatty acid and lipid composition of four marine fish larvae

• 1.1. Fatty acid and lipid class composition were determined in larvae of four marine species: Atlantic halibut (Hippoglossus hippoglossus L.), plaice (Pleuronectes platessa), cod (Gadus morhua) and turbot (Scophthalmus maximus) at hatching and prior to first feeding.
• 2.2. Total fatty acid content decreased in the four species with up to 50% reduction in one of the halibut groups. Docosahexanaoic acid (22:6 n-3) was especially utilized.
• 3.3. Low lipid utilization was found in turbot in relation to the other three species.
• 4.4. Water environmental temperature may explain some of the differences in the fatty acid utilization and the source of metabolic energy between cold water species (halibut, cod, and plaice) and temperate species (turbot), in the period from hatching to prior to first feeding.
• 5.5. Relative amounts of neutral lipids and phospholipids were similar in plaice, cod and halibut, approximately 25% and 75% of total lipids, respectively, and were approximately constant during the yolk-sac stage. Neutral lipids were dominant for turbot at hatching, accounting for 53–55% of the total lipids, while phospholipids predominated prior to first feeding, being 56–59%.
• 6.6. Phosphatidylcholine was catabolized in halibut, plaice and cod but not in turbot, while phosphatidylethanolamine tended to be synthesized in all four species.

     

Evidence of an egg-laying factor in the prostatic secretions of Helix aspersa müller

• 1.1. A high percentage (53%) of isolated snails injected with prostate gland homogenates lay eggs.
• 2.2. These egg masses consist of a few eggs which contain many nonviable oocytes.
• 3.3. Preliminary experiments suggest that an egg-laying factor may be present in prostatic secretions.
• 4.4. Snails bred in isolation from hatching, whether injected or not, occasionally lay viable eggs.
• 5.5. This observation shows that self-fertilization or parthenogenesis is, in fact, possible in Helix aspersa Müller.

     

Absorption and blood clearance of labelled carotenoids ([14C]astaxanthin, [3H]Canthaxanthin and [3H]zeaxanthin) in mature female rainbow trout (Oncorhynchus mykiss)

• 1.1. Using one force-fed meal, eight mature female rainbow trout received [¹⁴C]astaxanthin ([¹⁴C]Ax) with [³H]canthaxanthin ([³H]Cx; N = 3) or with [³H]zeaxanthin ([³H]Zx; N = 5).
• 2.2. Approximately 200 μl of blood were collected via caudal puncture every 24 hr for 4 days. After 96 hr, the fish were killed and pyloric caeca (P.C.) from the duodenal intestine (D.I.) section, ileal intestine (I.I.), and posterior intestine (P.I.) were dissected out.
• 3.3. In the blood, Ax levels were higher than Cx followed by Zx levels.
• 4.4. This corresponds to their respective absorption by the trout as was confirmed by their relative concentrations in P.C., I.I. and P.I.
• 5.5. However, blood clearance was similar for all three compounds. [¹⁴C]Phoenicoxanthin ([¹⁴C]Px) was detected as a reduced metabolite of [¹⁴C]Ax in all gut sections.

     

Phosphorylation in crayfish (Procambarus clarkii) eggs during hatching

• 1.1.|The high-energy phosphorylation metabolism in crayfish, Procambarus clarkii eggs during brooding and juvenile crayfish after hatching was studied by in vivo³¹P nuclear magnetic resonance (³¹P NMR) spectroscopy.
• 2.2.|Inorganic phosphoric acid (Pi) and adenosine-5′-triphosphate ATP(γ-,α-,β-) were detected in the dark brownish red eggs after oviposition.
• 3.3.|In orange unhatched eggs, only sugar phosphate (SP), Pi and resolved phosphometabolite from ATP were observed.
• 4.4.|Peaks of SP, Pi, arginine phosphate (Arg-P), and ATP (γ,α,β) appeared in larvae of crayfish after hatching (nauplius, zoea and juvenile crayfish).
• 5.5.|The high-energy phosphorylation metabolism changed to an anaerobic condition along with a decrease in the concentration of dissolved oxygen in fresh water.

     

Specific dynamic action (SDA) in the supralittoral isopod,Ligia pallasii: Identification of components of apparent SDA and effects of dietary amino acid quality and content on SDA

• 1.1. Specific Dynamic Action (SDA) effects of diet were investigated in the supralittoral isopod, Ligia pallasii, using defined chemical diets.
• 2.2. “Apparent SDA”, or the total rise in metabolic rate following a meal, was resolved in animals eating a nutritionally complete chemical diet into three components: 8% mechanical costs of moving food through the gut, 40% “excitement costs” due to investigator disturbance and presence of food, and 52% SDA.
• 3.3. Excitement costs in animals exposed to food but which chose not to eat showed non-significant variation between diets containing different levels of chemical nutrients, but were significantly less on a diet containing only cellulose and agar.
• 4.4. SDA increased with increasing concentration of amino acids in the diet.
• 5.5. Substitution of whole-protein casein for free amino acids in the diet had no significant SDA effect, while substitution of free amino acids in the ratio found in casein more than doubled the SDA effect.
• 6.6. Deletion of alanine from the diet caused no significant effect on SDA, while deletion of phenylalanine caused a highly significant elevation in SDA.

     

Effects of low temperatures on NAD-sorbitol dehydrogenase activity and morphogenesis in non-diapause eggs of the silkworm,Bombyx mori

• 1.1. The activity of NAD-sorbitol dehydrogenase (NAD-SDH; EC 1.1.1.14) and levels of sorbitol were examined in non-diapause eggs of the silkworm, Bombyx mori, exposed to temperatures of 20-0.5°C from 1 day after oviposition. The morphology of embryos in the cold-acclimated eggs and the hatching of eggs after transfer to 25°C were monitored.
• 2.2. Temperatures between 15 and 0.5°C retarded the development of NAD-SDH activity at a specific embryonic stage that was comparable to diapause, and sorbitol accumulated in the eggs.
• 3.3. With the appearance of NAD-SDH activity, sorbitol was converted into glycogen, just as it is in diapause eggs. The results indicate that NAD-SDH participates in the utilization of sorbitol rather than in its formation in non-diapause eggs.
• 4.4. Distinct effects of low temperatures on the morphological development of the embryos are also discussed.

     

γ-Glutamyltranspeptidase in echinoderm eggs and larvae: fertilization-dependent and developmentally-induced changes in specific activity

• 1.1. γ-Glutamyltranspeptidase is present in echinoderm eggs and larvae: in homogenates the level of activity is comparable to that of rat cerebral cortex.
• 2.2. In eggs of Lytechinus pictus, fertilization induces an early rapid and sustained (5 min–6 hr) 37% increase in the activity of γ-glutamyltranspeptidase in homogenate fractions.
• 3.3. Relative to these homogenate levels, the specific activity of γ-glutamyltranspeptidase are ≈60% lower in 40,000 g supernatant fractions and 2.7-fold higher in 40,000 g particulate fractions in both unfertilized and 15 min post-fertilized Lytechinus pictus eggs.
• 4.4. The subcellular distribution of γ-glutamyltranspeptidase is the same in both unfertilized and 15-min post-fertilized Lytechinus pictus eggs: 78% in 40,000 g particulate fractions, 22% in 40,000 g soluble fractions.
• 5.5. In both unfertilized and 15 min post-fertilized eggs of Lytechinus pictus the enzyme responds to heat (50 vs 37°C) by activation in a similar manner: 1.72- and 1.68-fold homogenates; 2.6- and 3.0-fold in supernatants; 1.97- and 1.90-fold in particulate fractions.
• 6.6. In homogenates of Pisaster ochraceous larvae, γ-glutamyltranspeptidase activity increases steadily during the course of larval development: relative to the low activity at day 5, activities exhibit an increase of 1.2-, 2.0-, 3.1- and 5.4-fold at days 10, 16, 22 and 28, respectively.

     

Photo-activation of respiration in the presence of CO in sperm of several marine invertebrates

• 1.1. The light irradiation at examined wavelengths between 360 and 630 nm enhanced the respiratory rate in the presence of CO in sperm, as well as eggs, of the sea urchins, Anthocidaris crassispina and Hemicentrotus pulcherrimus, the starfish, Asterina pectinifera, and the echiuroid, Urechis unicinctus.
• 2.2. The maximum peaks of stimulating effects of light irradiation on CO-insensitive respiration were found at wavelengths of 430, 530 and 570 nm in sperm of these species.
• 3.3. The respiration in the presence of CO was insensitive to light irradiation in sperm and eggs of the oyster, Crassostrea gigas and the tunicate, Ciona intestinaris.

     

The interaction of cytosolic epoxide hydrolase with chiral epoxides

• 1.1. The kinetic parameters of the cytosolic epoxide hydrolase were examined with two sets of spectrophotometric substrates. The (2S,3S)- and (2R,3R)-enantiomers of 4-nitrophenyl trans-2,3-epoxy-3-phenylpropyl carbonate had a K_mof 33 and 68 μm and a V_max of 16 and 27 μmol/min/mg, respectively. With the (2S,3S)- and (2R,3R)- enantiomers of 4-nitrophenyl trans-2,3-epoxy-3-(4-nitrophenyl)propyl carbonate, cytosolic epoxide hydrolase had a K_M of 8.0 and 15 μM and a V_max of 7.8 and 5.0 μmol/min/mg, respectively.
• 2.2. Glycidyl 4-nitrobenzoate had the lowest I₅₀ of the compounds tested in the glycidyl 4-nitrobenzoate series (I₅₀= 140 μM). The I₅₀ of the (2R)-enantiomer was 3.7-fold higher. The inhibitor with the lowest i₅₀ in the glycidol series, and the lowest I₅₀ of any compound tested, was (2S,3S)-3-(4-nitrophenyl)glycidol (I₅₀ = 13.0μM). It also showed the greatest difference in I₅₀ between the enantiomers (330-fold).
• 3.3. All enantiomers of glycidyl 4-nitrobenzoates and _trans-3-phenylglycidols gave differential inhibition of cytosolic epoxide hydrolase. However, neither the (S,S)-/(S)- or (R,R)-/(R)-enantiomer always had the lower I₅₀.
• 4.4. Addition of one or more methyl groups to either enantiomer of glycidyl 4-nitrobenzoate resulted in increased I₅₀. However, addition of a methyl group to C₂ of either enantiomer of 3-phenylglycidol resulted in a decreased I₅₀. Finally, when the hydroxyl group of trans-3-(4-nitrophenyl)glycidol was esterified the I₅₀ of the (2S,3S)- but not the (2R,3R)-enantiomer increased.