Masaki Kaneniwa

Mycosporine-Like Amino Acids Extracted from Scallop ( Patinopecten yessoensis ) Ovaries: UV Protection and Growth Stimulation Activities on Human Cells

Scallops (Patinopecten yessoensis) are extensively cultured and landed in Japan. During the processing of scallops, large amounts of internal organs and shells are discharged as industrial wastes. To reduce the burden on the environment, effective utilization and disposal methods of the wastes are required. Therefore, we have screened for useful materials in scallop internal organs, and found ultraviolet (UV) absorbing compounds from scallop ovaries. Based on UV absorption, electrospray ionization-mass spectrometry (ESI-MS), ESI-MS/MS, and nuclear magnetic resonance (NMR) spectra, three UV absorbing compounds were identified as mycosporine-like amino acids (MAAs): shinorine, porphyra-334 (P-334), and mycosporine-glycine. To investigate whether MAAs can act as a UV protector for human cells, we examined the protective effects of the three MAAs on human fibroblast cells from UV irradiation. All of the three examined MAAs protected the cells from UV-induced cell death. In particular, mycosporine-glycine had the strongest effect. Further, we found a promotion effect of MAAs on the proliferation of human skin fibroblast cells. From these results, it was found that the three MAAs isolated from scallop ovaries have a protective effect on human cells against UV light. MAAs have potential applications in cosmetics and toiletries as a UV protectors and activators of cell proliferation.

Fatty acids in crinoidea and ophiuroidea: Occurrence of all-cis-6,9,12,15,18,21-tetracosahexaenoic acid

The fatty acid compositions of lipids from two species of Crinoidea and two species of Ophiuroidea have been investigated with open-tubular gas chromatography. About 5–10% of tetracosahexaenoic acid was found in total fatty acids from all the samples, and the structure was determined as all-cis-6,9,12,15,18,21-tetracosahexaenoic acid [24∶6(n−3)] by13C-NMR of the methyl esters and mass spectrometric analyses of the methyl esters, the pyrrolidides and the ozonolysis products. The 24∶6(n−3) was concentrated in the polar lipids rather than neutral lipids. The n−3 hexaenoic structure suggested chain elongation of 22∶6(n−3) as the source.The 5-olefinic acids (5−18∶1, 5−20∶1, 5,11- and 5,13−20∶2) were low in Crinoidea (0.2–1.3%) but were present in higher levels (2.5–5.2%) in Ophiuroidea. Polyunsaturated acids found other than 24∶6(n−3) were 20∶4(n−6), 20∶5(n−3) and 22∶6(n−3) as major components and 16∶3(n−3), 18∶2(n−6), 18∶3(n−6), 18∶3(n−3), 18∶4(n−3), 20∶2(n−9), 20∶2(n−6), 20∶3(n−6), 20∶3(n−3), 21∶5(n−3) and 22∶5(n−3) as minor components in all the samples.

Fatty acids in echinoidea: Unusualcis-5-olefinic acids as distinctive lipid components in sea urchins

Open tubular gas liquid chromatographic (GLC) analyses of fatty acids from total lipids of 12 species of Echinoidea collected at several locations along the Pacific coast of Japan showed the same unusualcis-5-olefinic acids in all species, i.e.,cis-5-octadecenoic acid (5–18∶1),cis-5-eicosenoic acid (5–20∶1), all-cis-5,11- and 5,13-eicosadienoic acids (5,11- and 5,13–20∶2), allcis-5,11,14-eicosatrienoic acid (5,11,14–20∶3) and all-cis-5,11,14,17-eicosatetraenoic acid (5,11,14,17–20∶4). The structural analysis of partially purified 5,11,14,17–20∶4 was undertaken by reductive ozonolysis with GLC and gas chromatographic-mass spectrometric analyses of the products.13C-Nuclear magnetic resonance analyses of the totals and fractions of fatty acid methyl esters from the sea urchin lipids did not show any occurrence of fatty acids having an isolated olefinic bond in the 2, 3 or 4 positions. The 5-olefinic acids were concentrated on the polar lipids rather than neutral lipids. The branched and odd chain fatty acid contents of mud-feeding sea urchins were found to be relatively greater proportions of total fatty acids than in algae feeders.

Effect of tetracosahexaenoic acid on the content and release of histamine, and eicosanoid production in MC/9 mouse mast cell

Abstract6,9,12,15,18,21-Tetracosahexaenoic acid (24∶6n−3) was isolated from a brittle star, Ophiura sarsi Lütken, at>95% purity to evaluate its physiological functions. The effects of 24∶6n−3 on the production of leukotriene (LT)-related compounds such as LTB4, LTC4 and 5-hydroxyeicosatetraenoic acid, and the accumulation and release of histamine in an MC/9 mouse mast cell line were studied. We found that 24∶6n−3 could inhibit the antigen-stimulated production of LT-related compounds as well as other n−3 polyunsaturated fatty acids (PUFA) such as eicosapentaenoic acid (20∶5n−3) and docosahexaenoic acid (22∶6n−3), which are major n−3 PUFA in fish oils; 24∶6n−3 was also shown to reduce the histamine content in MC/9 cells at 25 μM (27% reduction from the control), and the effect was diminished with increase of the fatty acid concentration (up to 100 μM). These two n−3 PUFA, 20∶5n−3 and 22∶6n−3, also reduced the histamine content (16 and 20% reduction at 25 μM, respectively), whereas arachidonic acid (20∶4n−6) increased it (18% increase at 25 μM). Spontaneous- and antigen-induced release of histamine was not influenced with these PUFA (at 25 μM). Ionophore-stimulated release of histamine was suppressed by the PUFA (13,9,15, and 11% reduction with 20∶4n−6, 20∶5n−3, 22∶6n−3, and 24∶6n−3, respectively). The patterns of the effects of 24∶6n−3 on the synthesis of eicosanoids and histamine content were more similar to those of 22∶6n−3 than 20∶5n−3. From these results, 24∶6n−3 can be expected to have anti-inflammatory activity and antiallergic activities similar to those of 22∶6n−3.

Assimilation, Accumulation, and Metabolism of Dinophysistoxins (DTXs) and Pectenotoxins (PTXs) in the Several Tissues of Japanese Scallop Patinopecten yessoensis

Japanese scallops, Patinopecten yessoensis, were fed with the toxic dinoflagellate Dinophysis fortii to elucidate the relative magnitude of assimilation, accumulation, and metabolism of diarrhetic shellfish toxins (DSTs) and pectenotoxins (PTXs). Three individual scallops were separately exposed to cultured D. fortii for four days. The average cell number of D. fortii assimilated by each individual scallop was 7.7 × 105. Dinophysistoxin-1 (DTX1), pectenotoxin-2 (PTX2) and their metabolites were analyzed by liquid chromatography tandem mass spectrometry (LC/MS/MS) and the toxin content in individual tissues (digestive gland, adductor muscle, gill, gonad, mantle, and the others), feces and the seawater medium were quantified. Toxins were almost exclusively accumulated in the digestive gland with only low levels being detected in the gills, mantles, gonads, and adductor muscles. DTX1 and PTX2 were the dominant toxins in the D. fortii cells fed to the scallops, whereas the dominant toxins detected in the digestive gland of scallops were PTX6 and esterified acyl-O-DTX1 (DTX3). In other tissues PTX2 was the dominant toxin observed. The ratio of accumulated to assimilated toxins was 21%–39% and 7%–23% for PTXs and DTXs respectively. Approximately 54%–75% of PTX2 and 52%–70% of DTX1 assimilated by the scallops was directly excreted into the seawater mainly without metabolic transformation.

Autoxidation rates of 5-olefinic monoenoic and denoic fatty acids from sea urchin lipids and meadoefoam oils

Autoxidation rates of the 5-olefinic monoenoic and dienoic fatty acids from sea urchin lipids and meadow-foam oils were compared with those of normal monoenoic and dienoic fatty acids by gas-liquid Chromatographic determination of the unoxidized fatty acid methyl esters remaining through the autoxidation period. The fatty acids are classified into five groups shown below according to the oxidation rate of their methyl esters: I 5-olefinic monoenoic acids (c5-18:l, c5-20:l and c5-22:1), II normal monoenoic acids (c9-18:l, cll-18:l, c9-20:1, cl3-20:l and cl3-22:l), III 5-olefinic dienoic acids (c5,cll-20:2, c5,cl3-20:2 and c5,cl3-22:2), IV 7-olefinic dienoic acids (c7,cl3-22:2 and c7,cl5-22:2) and V normal dienoic acids (c6,c9-18:2, c9,cl2-18:2 and cll,cl4-20:2). The oxidation rates of these groups increased during autoxidation in order from I to V. These results show that the 5-olefinic monoenoic and dienoic acids are more stable to autoxidation than the normal monoenoic and dienoic acids, respectively. The higher stabilities of the 5-olefinic monoenoic and dienoic acids in organisms are shown from these results.

Enzymatic Hydrolysis of Lipids in Muscle of Fish and Shellfish During Cold Storage

In previous research (Olley and Lovern, 1960; Olley et al., 1962; Wu et al., 1974; Toyomizu et al., 1977; Hanaoka and Toyomizu, 1979; Ohshima and Koizumi, 1983; Ohshima et al., 1983a, Ohshima et al., 1983b, Ohshima et al., 1984a; Hwang and Regenstein, 1993; Aubourg et al., 1998; Ingemansson et al., 1995; Ben-Gigirey et al., 1999), enzymatic hydrolysis of lipids in fish muscle during cold storage has been reported in some lean and fatty fish such as cod, skipjack, carp, sardine, and rainbow trout. It is known that free fatty acids (FFA) accumulate in muscle lipids due to enzymatic hydrolysis of lipids, and the increase of FFA’s reduce the quality of the fish muscle (Dyer, 1951; Dyer and Fraser, 1959; Ohshima et al., 1984b). The functional fatty acid components such as EPA (20:5n-3) and DHA (22:6n-3) are also released from polar lipids or triacylglycerols to the free fatty acid fraction by hydrolysis, and it is presumed that the lipid hydrolysis affects their trophic value. The enzymatic hydrolysis was mainly caused by phospholipase in muscle, and (1997) partially purified phospholipase A1 from skipjack muscle. It is necessary to elucidate properties of enzymes for the prevention of enzymatic hydrolysis occurring in fish muscle. In this study, muscle of nine species of fish and shellfish (yellowtail, rainbow trout, bigeye tuna, chum salmon, skipjack, butterfly bream, Pacific cod, kuruma prawn, and scallop) were stored at −10°C for 30 days, and their lipid and fatty acid compositions analyzed before and after the storage for determination of the properties of enzymes concerned with lipid hydrolysis in muscles.

Relationships between quality parameters and content of glycerol galactoside and porphyra‐334 in dried laver nori Porphyra yezoensis

Glycerol galactoside (GG; floridoside + isofloridoside) and porphyra-334 (P-334) are contained in nori (Susabinori Porphyra yezoensis and Asakusanori Porphyra tenera). Glycerol galactoside has been found to have bifidogenic growth stimulator activity and P-334 is known to have ultraviolet-absorbing activity in the UVA region of sunlight. These substances have, respectively, potential for application to pre-biotic foods and in cosmetics as a sunscren.. In the present study, to investigate the relationships between GG and P-334 contents and the quality of nori, we measured the GG and P-334 contents with other components (total protein, chlorophyll-a, β-carotene and phycobillins) that are related to the quality of nori samples produced from different production areas and with different qualities. We found that the GG content was closely negatively correlated with the contents of other components, whereas P-334 was positively correlated with the other components. From these results, it is suggested that low-quality nori is a potential source of GG, and as a source for P-334, scraps of nori produced during nori processing should be suitable.

Prebiotic effect of glycerol galactoside isolated from color-faded nori in rats

Glycerol galactoside (GG; floridoside: 2-O-glycerol-α-d-galactopyranoside, and isofloridoside: 1-O-glycerol-α-d-galactopyranoside) is known to be a component of edible red seaweed nori. Recently, we have found that low-quality nori (color-faded nori), which has a low protein content, contains a large quantity of GG. From further studies, we have also found that GG has prebiotic characteristics in vitro. In this study, we evaluated the in vivo prebiotic activity of GG in rats. Dietary GG selectively increased the cecal Bifidobacterium count in rats. Other indices of prebiotics, such as pH of cecal content, organic acid concentrations, and fecal weight, also supported the existence of prebiotic activity of GG. The present data will also contribute to the development of a new method of utilizing color-faded nori as a health-promoting foodstuff.

Differences in compositions of free amino acids, nucleotide-related compounds, and fatty acids between sexes of the coconut crab Birgus latro in Okinawa, Southwest Japan

Despite the dietary utilization of the coconut crab Birgus latro throughout history, little is known about its nutritional composition and quality. This study assessed the differences between male and female coconut crabs caught in Okinawa, southwest Japan, in meat yield and compositions of free amino acids (FAA), nucleotide-related compounds (NRC), and fatty acids in muscle and hepatopancreas. The ratios of edible muscle and hepatopancreas to whole body weight were similar in males and females. Although concentrations of histidine and tryptophan in muscle and AMP in hepatopancreas differed in males and females, these differences were small, as were taste activities and equivalent umami concentrations, suggesting that the tastes of these tissues in males and females would be nearly identical. Fatty acid composition in muscle differed lightly between the sexes, with amounts of the 20:5n-3 and 22:6n-3 being significantly greater in males. However, the ratio of n-3 to n-6 polyunsaturated fatty acids and atherogenic and thrombogenic indices suggested that all lipids from both males and females are healthy for humans. There were no noticeable differences between the sexes in meat yield; FAA, NRC and fatty acid compositions; or nutritional qualities.