Researcher Profile and Settings

Affiliation

• Faculty of Fisheries Sciences　Marine Life Science　Marine Bioresources Chemistry

Job Title

• Professor

URL

https://fish-hokudai-bfmc.labby.jp/

J-Global ID

• 200901013802521530

Research Interests

• n-3 highly unsaturated fatty acids   antiinflammatory compounds   Astaxanthin   Fucoxanthin   EPA   DHA   カロテノイド   phospholipids   生活習慣病予防   Lipid Biochemistry   Food Chemistry   Molecular Nutrition   Fisheries Sciences   

Research Areas

• Life sciences / Aquaculture
• Life sciences / Molecular biology
• Life sciences / Food sciences
• Life sciences / Marine/Aquatic life sciences

Educational Organization

•  Bachelor's degree program　School of Fisheries Sciences
•  Master's degree program　Graduate School of Fisheries Sciences
•  Doctoral (PhD) degree program　Graduate School of Fisheries Sciences

Academic & Professional Experience

• 2017/04 - Today Hokkaido University Professor
• 2009 - 2017/03 北海道大学 水産科学研究科(研究院) 准教授

Association Memberships

• THE JAPANESE SOCIETY FOR MARINE BIOTECHNOLOGY   日本食品科学工学会   日本肥満学会   日本栄養食糧学会   脂質生化学研究会   脂質栄養学会   日本油化学会   日本農芸化学会   アメリカ油化学会   日本水産学会   

Research Activities

Published Papers

• Identification of a novel monocyclic carotenoid and prediction of its biosynthetic genes in Algoriphagus sp. oki45.

  Naoki Takatani, Takashi Maoka, Tomoo Sawabe, Fumiaki Beppu, Masashi Hosokawa

  Applied microbiology and biotechnology 108 (1) 102 - 102 2024/12 

  Bacteria belonging to the genus Algoriphagus have been isolated from various sources, such as Antarctic sea ice, seawater, and sediment, and some strains are known to produce orange to red pigments. However, the pigment composition and biosynthetic genes have not been fully elucidated. A new red-pigmented Algoriphagus sp. strain, oki45, was isolated from the surface of seaweed collected from Senaga-Jima Island, Okinawa, Japan. Genome comparison revealed oki45's average nucleotide identity of less than 95% to its closely related species, Algoriphagus confluentis NBRC 111222 T and Algoriphagus taiwanensis JCM 19755 T. Comprehensive chemical analyses of oki45's pigments, including 1H and 13C nuclear magnetic resonance and circular dichroism spectroscopy, revealed that the pigments were mixtures of monocyclic carotenoids, (3S)-flexixanthin ((3S)-3,1'-dihydroxy-3',4'-didehydro-1',2'-dihydro-β,ψ-caroten-4-one) and (2R,3S)-2-hydroxyflexixanthin ((2R,3S)-2,3,1'-trihydroxy-3',4'-didehydro-1',2'-dihydro-β,ψ-caroten-4-one); in particular, the latter compound was new and not previously reported. Both monocyclic carotenoids were also found in A. confluentis NBRC 111222 T and A. taiwanensis JCM 19755 T. Further genome comparisons of carotenoid biosynthetic genes revealed the presence of eight genes (crtE, crtB, crtI, cruF, crtD, crtYcd, crtW, and crtZ) for flexixanthin biosynthesis. In addition, a crtG homolog gene encoding 2,2'-β-hydroxylase was found in the genome of the strains oki45, A. confluentis NBRC 111222 T, and A. taiwanensis JCM 19755 T, suggesting that the gene is involved in 2-hydroxyflexixanthin synthesis via 2-hydroxylation of flexixanthin. These findings expand our knowledge of monocyclic carotenoid biosynthesis in Algoriphagus bacteria. KEY POINTS: • Algoriphagus sp. strain oki45 was isolated from seaweed collected in Okinawa, Japan. • A novel monocyclic carotenoid 2-hydroxyflexixanthin was identified from strain oki45. • Nine genes for 2-hydroxyflexixanthin biosynthesis were found in strain oki45 genome.
• Identification and tissue distribution of fucoxanthinol and amarouciaxanthin A fatty acid esters in fucoxanthin-fed mice

  Naoki Takatani, Kana Sakimura, Kodai Nagata, Fumiaki Beppu, Yumiko Yamano, Takashi Maoka, Masashi Hosokawa

  Food Chemistry 410 0308-8146 2023/06/01 

  Administered carotenoid fatty acid esters are thought to be hydrolyzed to their free forms and absorbed into the body, and information on the tissue distribution of carotenoid fatty acid esters has been limited. Fucoxanthin, a marine carotenoid, exhibits various health benefits, including anti-diabetic and anti-obesity effects. However, fucoxanthin metabolism in mammals remains unclear. Herein, we investigated the fatty acid esters of fucoxanthin metabolites, fucoxanthinol and amarouciaxanthin A, in the tissues of male C57BL/6J mice fed a fucoxanthin-containing diet for one week. Fucoxanthinol and amarouciaxanthin A-3-esters accumulated abundantly in the liver and epididymal white adipose tissue, respectively. These esters were less detectable in the serum and other tissues. Therefore, it is suggested that fucoxanthinol and amarouciaxanthin A are partially acylated in the liver and epididymal white adipose tissue after being transported through the body as their free forms. This study presents a novel carotenoid metabolic pathway in mammals.
• Dietary Combination of Fish Oil and Soy β-Conglycinin Inhibits Fat Accumulation and Reduces Blood Glucose Levels by Altering Gut Microbiome Composition in Diabetic/Obese KK-<i>A</i> ^<i>y</i> Mice

  Satoshi Wanezaki, Takumi Taniwaki, Junki Miyamoto, Masashi Hosokawa

  Journal of Oleo Science 72 (3) 303 - 312 1345-8957 2023
• Identification and tissue distribution of fucoxanthinol and amarouciaxanthin A fatty acid esters in fucoxanthin-fed mice.

  Naoki Takatani, Kana Sakimura, Kodai Nagata, Fumiaki Beppu, Yumiko Yamano, Takashi Maoka, Masashi Hosokawa

  Food chemistry 410 135318 - 135318 2022/12/28 

  Administered carotenoid fatty acid esters are thought to be hydrolyzed to their free forms and absorbed into the body, and information on the tissue distribution of carotenoid fatty acid esters has been limited. Fucoxanthin, a marine carotenoid, exhibits various health benefits, including anti-diabetic and anti-obesity effects. However, fucoxanthin metabolism in mammals remains unclear. Herein, we investigated the fatty acid esters of fucoxanthin metabolites, fucoxanthinol and amarouciaxanthin A, in the tissues of male C57BL/6J mice fed a fucoxanthin-containing diet for one week. Fucoxanthinol and amarouciaxanthin A-3-esters accumulated abundantly in the liver and epididymal white adipose tissue, respectively. These esters were less detectable in the serum and other tissues. Therefore, it is suggested that fucoxanthinol and amarouciaxanthin A are partially acylated in the liver and epididymal white adipose tissue after being transported through the body as their free forms. This study presents a novel carotenoid metabolic pathway in mammals.
• Monocaprin Enhances Bioavailability of Fucoxanthin in Diabetic/Obese KK-Ay Mice

  Kodai Nagata, Naoki Takatani, Fumiaki Beppu, Aya Abe, Etsuko Tominaga, Tomohisa Fukuhara, Makoto Ozeki, Masashi Hosokawa

  Marine Drugs 20 (7) 446 - 446 2022/07/07 [Refereed]
   

  Fucoxanthin is a marine carotenoid found in brown seaweeds and several microalgae. It has been reported that fucoxanthin has health benefits such as anti-obesity and anti-diabetic effects. To facilitate fucoxanthin applications in the food industry, it is important to improve its low bioavailability. We attempted the combined feeding of fucoxanthin-containing seaweed oil (SO) and monocaprin in a powder diet and analyzed the fucoxanthin metabolite contents in the liver, small intestine and serum of diabetic/obese KK-Ay mice. After 4 weeks of feeding with the experimental diets, the serum fucoxanthinol concentrations of the mice fed 0.2% SO and 0.5% monocaprin were higher than those of the 0.2% SO-fed mice. Furthermore, fucoxanthinol accumulation in the liver and small intestine tended to increase in a combination diet of 0.2% SO and 0.125–0.5% monocaprin compared with a diet of 0.2% SO alone, although amarouciaxanthin A accumulation was not different among the 0.2% SO-fed groups. These results suggest that a combination of monocaprin with fucoxanthin-containing SO is an effective treatment for improving the bioavailability of fucoxanthin.
• Preparation of Apoastaxanthinals and Evaluation of Their Anti-inflammatory Action against Lipopolysaccharide-Stimulated Macrophages and Adipocytes

  Naoki Takatani, Fumiaki Beppu, Yumiko Yamano, Takashi Maoka, Kazuo Miyashita, Masashi Hosokawa

  ACS OMEGA 2470-1343 2022/06 [Refereed]
   

  Apocarotenoids are carotenoid derivatives in which the polyene chain is cleaved via enzymatic or nonenzymatic action. They are found in animal tissues and carotenoid-containing foods. However, limited information on the biological functions of apocarotenoids is available. Here, we prepared apocarotenoids from astaxanthin via chemical oxidation and evaluated their anti-inflammatory action against macrophages and adipocytes. A series of astaxanthin-derived apoastaxanthinals, apo-11-, apo-15-, apo-14'-, apo-12'-, apo-10'-, and apo-8'astaxanthinals, were successfully characterized by chromatography and spectroscopic analysis. The apoastaxanthinals inhibited inflammatory cytokine production and mRNA expression against lipopolysaccharide-stimulated RAW 264.7 macrophages. Apoastaxanthinals suppressed interleukin-6 overexpression in an in vitro model with macrophages and adipocytes in the following cultures: (1) contact coculture of 3T3-L1 adipocytes and RAW264.7 macrophages and (2) 3T3-L1 adipocytes in a RAW264.7-derived conditioned media. These results indicate that the apoastaxanthinals have the potential for regulation of adipose tissue inflammation observed in obesity.
• Reduction of serum lipid levels in diabetic obese KK-A ^y mice using monoalkyldiacylglycerol-rich oil derived from Berryteuthis magister

  Masaki Kakimoto, Naoki Takatani, Masashi Hosokawa, Fumiaki Beppu

  Fisheries Science 0919-9268 2022 

  Monoalkyldiacylglycerol (MADG), an ether analog of triacylglycerols, is abundant in some marine animals such as deep-sea sharks and echinoderms. The present study aims to investigate the effects of an MADG-rich neutral lipid derived from Berryteuthis magister (MNB) on the lipid metabolism of diabetic/obese KK-Ay mice. Male mice (5 weeks old) were fed soybean oil (control group), MNB, or fish oil (FO) which contain the same amount of EPA and DHA in the form of triacylglycerols. MNB or FO supplementation resulted in a decrease in adipose tissue weights and blood glucose levels compared to the control mice. Interestingly, serum lipid levels in the MNB group were significantly lower than those of the control and FO groups. Although there was no difference in EPA and DHA levels in the tissues between the FO and MNB groups, a decrease in the number of lipoprotein particles was characteristically observed in the mice fed with MNB. Our results suggest that the MADG structure-dependent effect on lipoprotein synthesis and/or secretion partly accounted for the reduced serum lipids in KK-Ay mice by dietary MNB. This study advances the understanding of MADG as a nutraceutical and provides insights into the role of ether lipids in lipoprotein metabolism that can contribute to the development of prevention and treatment strategies for hyperlipidemia.
• n-3 Polyunsaturated fatty acid-enriched phosphatidylglycerol suppresses inflammation in RAW264.7 cells through Nrf2 activation via alteration of fatty acids in cellular phospholipids

  Liping Chen, Fumiaki Beppu, Naoki Takatani, Kazuo Miyashita, Masashi Hosokawa

  FISHERIES SCIENCE 87 (5) 727 - 737 0919-9268 2021/09 [Refereed]
   

  Phosphatidylglycerol (PG) is a naturally occurring phospholipid. The immunomodulatory activity of PG has been reported to be dependent on its fatty acid composition. In this study, the anti-inflammatory effect of n-3 polyunsaturated fatty acid-enriched PG (n-3 PUFA-PG) was investigated on macrophage-like RAW264.7 cells stimulated by lipopolysaccharide (LPS). n-3 PUFA-PG binding eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) significantly suppressed interleukin-6, interleukin-1 beta and nitric oxide production by RAW264.7 cells. Notably, the suppressive effect of n-3 PUFA-PG on inflammatory factor production was more potent compared to n-3 PUFA-phosphatidylcholine (PC) and soybean PG. n-3 PUFA-PG treatment markedly increased EPA and DHA in intracellular PC and phosphatidylethanolamine and decreased 18:1n-9 through downregulation of fatty acid synthase and stearoyl-CoA desaturase mRNA expression compared to n-3 PUFA-PC treatment. Furthermore, n-3 PUFA-PG enhanced nuclear factor erythroid E2-related factor (Nrf2) levels in the nucleus and increased heme oxygenase-1 mRNA expression. N-acetyl-L-cysteine, which is an inhibitor against Nrf-2 activation, alleviated the induction of HO-1 mRNA and anti-inflammatory effect of n-3 PUFA-PG. These data suggest that n-3 PUFA-PG exhibits anti-inflammatory effects on LPS-stimulated RAW264.7 cells through Nrf2 activation by altering cellular fatty acids.
• Seco-type β-Apocarotenoid Generated by β-Carotene Oxidation Exerts Anti-inflammatory Effects against Activated Macrophages.

  Naoki Takatani, Fumiaki Beppu, Yumiko Yamano, Takashi Maoka, Masashi Hosokawa

  Journal of oleo science 70 (4) 549 - 558 2021/04/02 [Refereed]
   

  β-Apocarotenoids are the cleavage products of β-carotene. They are found in plants, carotenoid-containing foods, and animal tissues. However, limited information is available regarding the health benefits of β-apocarotenoids. Here, we prepared seco-type β-apocarotenoids through the chemical oxidation of β-carotene and investigated their anti-inflammatory effects against activated macrophages. Oxidation of β-carotene with potassium permanganate produced seco-β-apo-8'-carotenal, in which one end-group formed an "open" β-ring and the other was cleaved at the C-7',8' position. In lipopolysaccharide-stimulated murine macrophage-like RAW264.7 cells, seco-β-apo-8'-carotenal inhibited the secretion and mRNA expression of inflammatory mediators such as nitric oxide, interleukin (IL)-6 and IL-1β, and monocyte chemoattractant protein-1. Furthermore, seco-β-apo-8'-carotenal suppressed phosphorylation of c-Jun N-terminal kinase and the inhibitor of nuclear factor (NF)-κB as well as the nuclear accumulation of NF-κB p65. Notably, since seco-β-apo-8'-carotenal exhibited remarkable anti-inflammatory activity compared with β-apo-8'-carotenal, its anti-inflammatory action could depend on the opened β-ring structure. These results suggest that seco-β-apo-8'-carotenal has high potential for the prevention of inflammation-related diseases.
• Effects of Fucoxanthin on the Inhibition of Dexamethasone-Induced Skeletal Muscle Loss in Mice

  Maki Yoshikawa, Masashi Hosokawa, Kazuo Miyashita, Hoyoku Nishino, Takeshi Hashimoto

  NUTRIENTS 13 (4) 2021/04 [Refereed]
   

  Fucoxanthin (Fx) has preventive effect against muscle atrophy and myotube loss in vitro, but it has not yet been examined in vivo. Therefore, we aimed to investigate the effect of Fx on dexamethasone (Dex)-induced muscle atrophy and fat mass in mice. ICR mice were fed with Fx diets from 2 weeks before Dex treatment to the end of the study. Muscle atrophy was induced in the mice by oral administration of Dex. Body weight was significantly lower by Dex treatment. Visceral fat mass in the Fx-treated group were significantly lower than those in the control group. The Dex-induced decrease in tibialis anterior muscle mass was ameliorated by Fx treatment. Fx treatment significantly attenuated muscle lipid peroxidation compared with the control and Dex-treated groups. The phosphorylation of AMPK was significantly higher in the Dex-treated group than in the control group. The expression of cytochrome c oxidase (COX) IV was significantly higher in the Fx-treated group than in the control group. These results suggest that Fx may be a beneficial material to prevent muscle atrophy in vivo, in addition to the effect of fat loss.
• Health-Promoting Functions of the Marine Carotenoid Fucoxanthin

  Masashi Hosokawa

  CAROTENOIDS: BIOSYNTHETIC AND BIOFUNCTIONAL APPROACHES 1261 273 - 284 0065-2598 2021 [Not refereed][Invited]
   

  Fucoxanthin (Fx) is a marine carotenoid found in brown seaweeds and several marine microalgae. Recent studies have reported that dietary Fx exhibits many health-promoting functions such as anti-obesity and anti-diabetic effects in animal experiments. A human clinical trial of Fx showed high potential on reduction of body weight and fat content. The anti-obesity effect of Fx is due to several mechanisms, which include the suppression of adipocyte differentiation, anti-inflammation, and uncouple protein 1 induction in white adipose tissue. Furthermore, Fx reduced blood glucose level and improved insulin resistance through the regulation of adipokine mRNA expressions. In this chapter, we reviewed health beneficial effects and safety of Fx and discussed their molecular mechanisms.
• Identification of Paracentrone in Fucoxanthin-Fed Mice and Anti-Inflammatory Effect against Lipopolysaccharide-Stimulated Macrophages and Adipocytes

  Naoki Takatani, Daisuke Taya, Ami Katsuki, Fumiaki Beppu, Yumiko Yamano, Akimori Wada, Kazuo Miyashita, Masashi Hosokawa

  MOLECULAR NUTRITION & FOOD RESEARCH 65 (2) e2000405  1613-4125 2021/01 [Refereed]
   

  Scope Fucoxanthin is converted to fucoxanthinol and amarouciaxanthin A in the mouse body. However, further metabolism such as cleavage products (i.e., apocarotenoids) remains unclear. The fucoxanthin-derived apocarotenoid in vivo is investigated and the anti-inflammatory effect of apocarotenoids with fucoxanthin partial structure such as allenic bond and epoxide residue against activated macrophages and adipocytes in vitro is evaluated.Methods and Results LC-MS analysis indicates the presence of paracentrone, a C-31-allenic-apocarotenoid, in white adipose tissue of diabetic/obese KK-A(y) and normal C57BL/6J mice fed 0.2% fucoxanthin diet for 1 week. In lipopolysaccharide-activated RAW264.7 macrophages, paracentrone as well as C-26- and C-28-allenic-apocarotenoids suppresses the overexpression of inflammatory factors. Further, apo-10 '-fucoxanthinal, a fucoxanthin-derived apocarotenoid which retained epoxide residue, exhibits a most potent anti-inflammatory activity through regulating mitogen-activated protein kinases and nuclear factor-kappa B inflammatory signal pathways. In contrast, beta-apo-8 '-carotenal without allenic bond and epoxide residue lacks suppressed inflammation. In 3T3-L1 adipocytes, paracentrone, and apo-10 '-fucoxanthinal downregulate the mRNA expression of proinflammatory mediators and chemokines induced by co-culture with RAW264.7 cells.Conclusion Dietary fucoxanthin accumulates as paracentrone as well as fucoxanthinol and amarouciaxanthin A in the mouse body. Allenic bond and epoxide residue of fucoxanthin-derived apocarotenoids have pivotal roles for anti-inflammatory action against activated macrophages and adipocytes.
• Black chokeberry (Aronia melanocarpa) juice residue and its ethanol extract decrease serum lipid levels in high‑fat diet‑fed C57BL/6J mice

  Nana Mikami, Yukie Hosotani, Tomoko Saso, Tomoki Ohta, Kazuo Miyashita, Masashi Hosokawa

  International Journal of Functional Nutrition 1 (2) 2634-7989 2020/10/30 [Refereed]
  
• Ecogenomics of the Marine Benthic Filamentous Cyanobacterium Adonisia

  Juline M. Walter, Felipe H. Coutinho, Luciana Leomil, Paulo I. Hargreaves, Mariana E. Campeao, Veronica V. Vieira, Beatriz S. Silva, Giovana O. Fistarol, Paulo S. Salomon, Tomoo Sawabe, Sayaka Mino, Masashi Hosokawa, Hideaki Miyashita, Fumito Maruyama, Marcel C. van Verk, Bas E. Dutilh, Cristiane C. Thompson, Fabiano L. Thompson

  MICROBIAL ECOLOGY 80 (2) 249 - 265 0095-3628 2020/08 [Refereed][Not invited]
   

  Turfs are among the major benthic components of reef systems worldwide. The nearly complete genome sequences, basic physiological characteristics, and phylogenomic reconstruction of two phycobiliprotein-rich filamentous cyanobacteria strains isolated from turf assemblages from the Abrolhos Bank (Brazil) are investigated. Both Adonisia turfae CCMR0081(T) (= CBAS 745(T)) and CCMR0082 contain approximately 8 Mbp in genome size and experiments identified that both strains exhibit chromatic acclimation. Whereas CCMR0081(T) exhibits chromatic acclimation type 3 (CA3) regulating both phycocyanin (PC) and phycoerythrin (PE), CCMR0082 strain exhibits chromatic acclimation type 2 (CA2), in correspondence with genes encoding specific photosensors and regulators for PC and PE. Furthermore, a high number and diversity of secondary metabolite synthesis gene clusters were identified in both genomes, and they were able to grow at high temperatures (28 degrees C, with scant growth at 30 degrees C). These characteristics provide insights into their widespread distribution in reef systems.
• Bioactive significance of fucoxanthin and its effective extraction

  Kazuo Miyashita, Fumiaki Beppu, Masashi Hosokawa, Xiaoyong Liu, Shuzhou Wang

  BIOCATALYSIS AND AGRICULTURAL BIOTECHNOLOGY 26 2020/07 [Refereed]
   

  Fucoxanthin (Fx) is a major carotenoid present in the chloroplasts of brown seaweeds and some kinds of microalgae. Because of the beneficial health effects of Fx, much attention has been focused on the considerable potential and promising applications of this compound in human health. Among the wide variety of biological activities of Fx, the interest has been mainly focused on its antiobesity effect, which is primarily based on the upregulation of thermogenesis through uncoupling protein 1 expression. Therefore, much effort has been made on the exploration for better Fx sources from different kinds of brown seaweeds and microalgae. The effective method to separate Fx has also attracted the interest of numerous researchers. Fx is conventionally extracted using organic solvents; however, solvents of petroleum origin are now strictly regulated worldwide. Thus, there is a growing demand for using greener, bio-based and renewable solvents for the extraction. From this viewpoint, edible oils may be promising green solvents. Edible oils can reduce the energy consumption in the extraction process and produce a barrier against Fx degradation. The edible oil extracts from brown seaweeds can be directly used as functional food materials containing Fx, without the need for purification.
• Preparation of Phosphatidyl-panthenol by phospholipase D-mediated transphosphatidylation and its anti-inflammatory activity on macrophage-like RAW264.7 cells

  Yukihiro Yamamoto, Kounosuke Suzuri, Tasuku Kunii, Hideyuki Kurihara, Kazuo Miyashita, Masashi Hosokawa

  BIOCATALYSIS AND AGRICULTURAL BIOTECHNOLOGY 26 101629 - 101629 1878-8181 2020/07 [Refereed][Not invited]
   

  Panthenol is known to be pro-vitamin B-5 and show several functions such as wound-healing, anti-oxidative, and anti-inflammatory activities. In this study, panthenol was phosphatidylated via phospholipase D (PLD)-mediated transphosphatidylation and its anti-inflammatory effects were investigated on macrophage-like RAW264.7 cells. By PLD-mediated transphosphatidylation from phosphatidylcholine (PC), hydroxy group of p1 position in panthenol molecule was alternatively formed phosphate ester bond even though panthenol has three of hydroxy groups in the molecule. The yield was 96 mol% under optimum reaction conditions: 50 umol of PC, 1.6 U of PLD, 500 umol of panthenol, 1.6 mL/1.6 mL of ethyl acetate/0.2 M acetate buffer (pH 5.6) ratio, 10 mM of CaCl2 in the buffer, 37 degrees C for 24 h. Phosphatidyl-panthenol (P-panthenol) suppressed mRNA expression level of pro-inflammatory mediators such as IL-6, IL-1 beta, TNF-alpha and COX-2 more than precursor compounds, PC and panthenol, in activated RAW264.7 cells. P-panthenol also suppressed protein secretion levels of IL-6 and IL-1 beta in the culture medium of RAW264.7 cells. These results demonstrate that P-panthenol, which is easily synthesized in high yield, is potential compound for novel anti-inflammatory phospholipid.
• Fucoxanthin inhibits hepatic oxidative stress, inflammation, and fibrosis in diet-induced nonalcoholic steatohepatitis model mice

  Naoki Takatani, Yuka Kono, Fumiaki Beppu, Yuko Okamatsu-Ogura, Yumiko Yamano, Kazuo Miyashita, Masashi Hosokawa

  BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS 528 (2) 305 - 310 0006-291X 2020/07 [Refereed][Not invited]
   

  Nonalcoholic steatohepatitis (NASH) is associated with hepatocyte injury, excessive oxidative stress, and chronic inflammation in fatty liver, and can progress to more severe liver diseases, such as cirrhosis and hepatocellular carcinoma. However, currently there are no effective therapies for NASH. Marine carotenoid, fucoxanthin (Fx), abundant in brown seaweeds, has variable biological properties, such as anticancer, anti-inflammatory, anti-oxidative and anti-obesity. However, the effect of Fx on the development of NASH has not been explored. We investigated the protective effects of Fx in diet-induced NASH model mice fed choline-deficient L-amino acid-defined high fat diet (CDAHFD). Fx administration significantly attenuated liver weight gain and hepatic fat accumulation, resulting in the alleviation of hepatic injury. Furthermore, the Fx-fed mice, not only exhibited reduced hepatic lipid oxidation, but also decreased mRNA expression levels of inflammation and infiltration-related genes compared to that of the CDAHFD-fed mice. Moreover, fucoxanthinol and amarouciaxanthin A, two Fx metabolites exerted anti-inflammatory effects in the liver via inhibiting the chemokine production in hepatocytes. In case of fibrosis, one of the features of advanced NASH, the expression of fibrogenic factors including activatedhepatic stellate cell marker was significantly decreased in the liver of Fx-fed mice. Thus, the present study elucidated that dietary Fx not only inhibited hepatic oxidative stress and inflammation but also prevented early phase of fibrosis in the diet-induced NASH model mice. (C) 2020 Elsevier Inc. All rights reserved.
• Nutraceutical characteristics of the brown seaweed carotenoid fucoxanthin

  Kazuo Miyashita, Fumiaki Beppu, Masashi Hosokawa, Xiaoyong Liu, Shuzhou Wang

  ARCHIVES OF BIOCHEMISTRY AND BIOPHYSICS 686 108364 - 108364 0003-9861 2020/06 [Refereed][Not invited]
   

  Fucoxanthin (Fx), a major carotenoid found in brown seaweed, is known to show a unique and wide variety of biological activities. Upon absorption, Fx is metabolized to fucoxanthinol and amarouciaxanthin, and these metabolites mainly accumulate in visceral white adipose tissue (WAT). As seen in other carotenoids, Fx can quench singlet oxygen and scavenge a wide range of free radicals. The antioxidant activity is related to the neuroprotective, photoprotective, and hepatoprotective effects of Fx. Fx is also reported to show anti-cancer activity through the regulation of several biomolecules and signaling pathways that are involved in either cell cycle arrest, apoptosis, or metastasis suppression. Among the biological activities of Fx, anti-obesity is the most well-studied and most promising effect. This effect is primarily based on the upregulation of thermogenesis by uncoupling protein 1 expression and the increase in the metabolic rate induced by mitochondrial activation. In addition, Fx shows anti-diabetic effects by improving insulin resistance and promoting glucose utilization in skeletal muscle.
• Fucoxanthinol attenuates oxidative stress-induced atrophy and loss in myotubes and reduces the triacylglycerol content in mature adipocytes

  Maki Yoshikawa, Masashi Hosokawa, Kazuo Miyashita, Takashi Fujita, Hoyoku Nishino, Takeshi Hashimoto

  MOLECULAR BIOLOGY REPORTS 47 (4) 2703 - 2711 0301-4851 2020/04 [Refereed][Not invited]
   

  The combination of sarcopenia and obesity (i.e., sarcopenic obesity) is more strongly associated with disability and metabolic/cardiovascular diseases than obesity or sarcopenia alone. Therefore, countermeasures that simultaneously suppress fat gain and muscle atrophy to prevent an increase in sarcopenic obesity are warranted. The aim of this study was to investigate the simultaneous effects of fucoxanthinol (FXOH) on fat loss in mature adipocytes and the inhibition of atrophy and loss in myotubes induced by oxidative stress. C2C12 myotubes were treated with FXOH for 24 h and further incubated with hydrogen peroxide (H2O2) for 24 h. The area of myosin heavy chain-positive myotubes and the ROS concentration were measured. Mature 3T3-L1 adipocytes were treated with FXOH for 72 h. The triacylglycerol (TG) content and glycerol and fatty acid (FA) release were biochemically measured. The myotube area was smaller in H2O2-treated cells than that in control cells. However, FXOH protected against the H2O2-induced decreases in myotube area. Further, the ROS concentration was significantly higher in the FXOH-treated cells compared with that in the control cells, although it was significantly lower than that in the H2O2-treated cells. On the other hand, in the mature adipocytes, the TG content was significantly decreased by FXOH treatment compared to that in the control. Moreover, FXOH treatment significantly increased glycerol and FA release compared with that of the control. These results suggest that FXOH inhibits H2O2-induced atrophy and loss in myotubes and activates lipolysis and decreases the TG content in mature adipocytes. Accordingly, FXOH has the potential to exert anti-sarcopenic obesity effects.
• Effective extraction of carotenoids from brown seaweeds and vegetable leaves with edible oils

  Kazuyoshi Teramukai, Shusuke Kakui, Fumiaki Beppu, Masashi Hosokawa, Kazuo Miyashita

  INNOVATIVE FOOD SCIENCE & EMERGING TECHNOLOGIES 60 1466-8564 2020/03 [Refereed]
   

  Factors affecting carotenoid extraction with edible oils were examined using edible brown seaweed, Sargassum horneri, as main sample. The results indicate that drying was essential to extract fucoxanthin (Fx) from S. horneri and physical (boiling) and chemical (acid/alkali) pretreatment of the wet sample increased the extraction rate of Fx. Additionally, more Fx was found from the dried S. horneri powder with a smaller particle size. The extraction rate of Fx is affected by the extraction temperature and time, showing that the effective extraction would be obtained at 50 degrees C within 12 hr extraction. Among the oils used, short-chain (C4 and C6) triacylglycerol (TAG) (SCT) and medium-chain (C8) TAG (MCT) could extract more Fx from S. horneri and more beta-carotene and lutein from spinach and olive leaves. The relatively lower viscosity of SCT and MCT would be the most likely reason for the higher extraction rates of both TAGs.
• Spirulina Lipids Alleviate Oxidative Stress and Inflammation in Mice Fed a High-Fat and High-Sucrose Diet

  Yuhong Yang, Lei Du, Masashi Hosokawa, Kazuo Miyashita

  MARINE DRUGS 18 (3) 2020/03 [Refereed][Not invited]
   

  High-fat and high-sucrose diet (HFHSD)-induced obesity leads to oxidative stress and chronic inflammatory status. However, little is known about the beneficial effects of total lipids extracted from Spirulina. Hence, in the present study, Spirulina lipids were extracted with chloroform/methanol (SLC) or ethanol (SLE) and then their effects on oxidative stress and inflammation in the mice fed a HFHSD were investigated. The results show that the major lipid classes and fatty acid profiles of SLC and SLE were almost similar, but the gamma-linolenic acid (GLA) and carotenoid contents in SLE was a little higher than that in SLC. Dietary 4% SLC or SLE for 12 weeks effectively decreased the hepatic lipid hydroperoxide levels as well as increased the activities and mRNA levels of antioxidant enzymes in the mice fed a HFHSD. In addition, supplementation with SLC and SLE also markedly decreased the levels of serum pro-inflammatory cytokines and the mRNA expression of pro-inflammatory cytokines in the liver and epididymal white adipose tissue of mice fed a HFHSD, and the effects of SLC and SLE were comparable. These findings confirm for the first time that dietary Spirulina lipids could alleviate HFHSD-induced oxidative stress and inflammation.
• Preparation of n-3 Polyunsaturated Phosphatidylglycerol from Salmon Roe Lipids by Phospholipase D and In Vitro Digestion

  Liping Chen, Fumiaki Beppu, Kazuo Miyashita, Masashi Hosokawa

  EUROPEAN JOURNAL OF LIPID SCIENCE AND TECHNOLOGY 122 (2) 1438-7697 2020/02 [Refereed]
   

  Phosphatidylglycerol (PG) is a highly functional phospholipid (PL), which has many physiological functions. However, naturally occurring PG binding n-3 polyunsaturated fatty acid (n-3 PUFA) is low in content, resulting in a scarcity of industrial bio-resources of n-3 PUFA enriched PG. The current study investigates the preparation of salmon roe PG (SRPG) from three types of salmon roe lipids and glycerol via phospholipase D (PLD)-mediated transphosphatidylation. The yields of SRPG obtained from salmon roe total lipid (SRTL) and salmon roe PL (SRPL) are higher than those obtained from purified salmon roe phosphatidylcholine (SRPC) in aqueous system. Following a 24 h reaction with 0.75 U PLD, SRTL, and SRPL yield up to 96.4 mol% and 96.7 mol% SRPG, respectively. In addition, more fatty acids are released from synthesized SRPG via hydrolysis by pancreatic enzymes than from SRPC and soybean PC in in vitro digestion model. Fatty acids at the sn-2 position of SRPG are completely liberated by 0.04 U of phospholipase A(2) (PLA(2)) during a 6 h reaction, whereas fatty acids of SRPC are partially unhydrolyzed even after a 24 h reaction. Our results suggest that SRPG converted from salmon lipids by PLD is a functional PL with high bioavailability of n-3 PUFAs. Practical Applications: Phosphatidylglycerol rich in n-3 PUFAs is prepared from salmon roe lipids (SRPG) catalyzed by PLD. The SRPG yields reach 96.4 mol% and 96.7 mol% of phosphatidylcholine contained in SRTL and SRPL, respectively, in aqueous reaction system. Fatty acids rich in n-3 PUFAs at sn-2 position of prepared SRPG are rapidly liberated by PLA(2) in an in vitro digestion model.
• Effect of Spirulina lipids on high-fat and high-sucrose diet induced obesity and hepatic lipid accumulation in C57BL/6J mice

  Yuhong Yang, Lei Du, Masashi Hosokawa, Kazuo Miyashita

  JOURNAL OF FUNCTIONAL FOODS 65 1756-4646 2020/02 [Refereed][Not invited]
   

  Obesity is characterized by abnormal or excessive fat accumulation that impairs health. In this study, the effects of Spirulina lipids (SL) and fish oil (FO) on obesity and hepatic lipid accumulation in high-fat and high-sucrose diet induced obese C57BL/6J mice were evaluated and compared. After 12 weeks of feeding, 4% gamma-linolenic acid-enriched SL supplementation effectively reduced the gain of body weight, total white adipose tissue (WAT) weight and hepatic lipid levels in obese mice. Although dietary 4% FO also tended to suppress the increases of body weight and WAT weight in mice, no statistical difference was observed. The suppression of hepatic lipid accumulation by SL should be through regulating sterol regulatory element binding protein-1c (SREBP-1c), SREBP-2-mediated triacylglycerol and cholesterol synthesis pathway as well as via enhancement of peroxisome proliferator-activated receptor alpha (PPAR alpha)-mediated beta-oxidation. These findings firstly confirmed that SL could alleviate diet-induced obesity and hepatic lipid accumulation.
• 海洋性カロテノイドの健康機能性に関する最近の動向

  高谷 直己, 細川 雅史

  日本食品安全協会会誌 15 78 - 85 2020 [Not refereed]
  
• Total Lipids Content, Lipid Class and Fatty Acid Composition of Ten Species of Microalgae

  Yuhong Yang, Lei Du, Masashi Hosokawa, Kazuo Miyashita

  JOURNAL OF OLEO SCIENCE 69 (10) 1181 - 1189 1345-8957 2020 [Refereed]
   

  Microalgae is a potential producer of functional lipids such as n-3 polyunsaturated fatty acids (PUFA) and fucoxanthin. In the present study, lipids from ten microalgae species were analyzed especially focusing on the fucoxanthin, lipid and fatty acid compositions. The study revealed a remarkable variation in total lipids content, fucoxanthin content, lipid class composition and n-3 PUFA content in individual species, although they belong to the same genus. Among microalgac examined, Pavlova lutheri contained the highest total lipids content (313.59 mg g(-1) dry weight) and considerable amount of fucoxanthin (3.13 mg g(-1) dry weight). It also had the highest level (28.01%) of total n-3 PUFA with high level of eicosapentaenoic acid (EPA) (17.76%) and docosahexaenoic acid (DHA) (7.61%). The highest fucoxanthin content (5.19 mg g(-1) dry weight) was observed in Chaetoceros gracilis. C gracilis also contained relatively high level of total lipids (228.87 mg g(-1). dry weight) and 10.67% EPA. The results also demonstrated that Nannochloropsis oculata contained the greatest amount of EPA (26.21%), while Isochrysis galbana had the highest level of DHA (8.76%). And both microalgae contained 1.71 and 4.44 mg g(-1) dry weight fucoxanthin, respectively. Microalgal lipids containing abundant fucoxanthin and n-3 PUFA such as EPA and DHA in the present study will be used as nutraceutical lipids with great commercial potential.
• The Effect of n-3 PUFA Binding Phosphatidylglycerol on Metabolic Syndrome-Related Parameters and n-3 PUFA Accretion in Diabetic/Obese KK-A(y) Mice

  Liping Chen, Naoki Takatani, Fumiaki Beppu, Kazuo Miyashita, Masashi Hosokawa

  NUTRIENTS 11 (12) 2019/12 [Refereed][Not invited]
   

  n-3 Polyunsaturated fatty acid binding phospholipids (n-3 PUFA-PLs) are known to be potent carriers of n-3 PUFAs and provide health benefits. We previously prepared n-3 PUFA binding phosphatidylglycerol (n-3 PUFA-PG) by phospholipase D-mediated transphosphatidylation. Because PG has excellent emulsifiability, n-3 PUFA-PG is expected to work as a functional molecule with properties of both PG and n-3 PUFAs. In the present study, the health benefits and tissue accretion of dietary n-3 PUFA-PG were examined in diabetic/obese KK-A(y) mice. After a feeding duration over 30 days, n-3 PUFA-PG significantly reduced the total and non-HDL cholesterols in the serum of diabetic/obese KK-A(y) mice. In the mice fed n-3 PUFA-PG, but not n-3 PUFA-TAG, hepatic lipid content was markedly alleviated depending on the neutral lipid reduction compared with the SoyPC-fed mice. Further, the n-3 PUFA-PG diet increased eicosapentaenoic acid and docosahexaenoic acid (DHA) and reduced arachidonic acid in the small intestine, liver, perirenal white adipose tissue, and brain, and the ratio of the n-6 PUFAs to n-3 PUFAs in those tissues became lower compared to the SoyPC-fed mice. Especially, the DHA level was more significantly elevated in the brains of n-3 PUFA-PG-fed mice compared to the SoyPC-fed mice, whereas n-3 PUFA-TAG did not significantly alter DHA in the brain. The present results indicate that n-3 PUFA-PG is a functional lipid for reducing serum and liver lipids and is able to supply n-3 PUFAs to KK-A(y) mice.
• Variation in Lipid Components from 15 Species of Tropical and Temperate Seaweeds

  Eko Susanto, A. Suhaeli Fahmi, Masashi Hosokawa, Kazuo Miyashita

  MARINE DRUGS 17 (11) 2019/11 [Refereed][Not invited]
   

  The present study describes the variation in lipid components from 15 species of seaweeds belonging to the Chlorophyta, Ochrophyta, and Rhodophyta phyla collected in tropical (Indonesia) and temperate (Japan) areas. Analyses were performed of multiple components, including chlorophylls, carotenoids, n-3 and n-6 polyunsaturated fatty acids (PUFAs), and alpha tocopherol (alpha-Toc). Chlorophyll (Chl) and carotenoid contents varied among phyla, but not with the sampling location. Chl a and b were the major chlorophylls in Chlorophyta. Chl a and Chl c were the main chlorophylls in Ochrophyta, while Chl a was the dominant chlorophylls in Rhodophyta. beta-Carotene and fucoxanthin were detected as major seaweed carotenoids. The former was present in all species in a variety of ranges, while the latter was mainly found in Ochrophyta and in small quantities in Rhodophyta, but not in Chlorophyta. The total lipids (TL) content and fatty acids composition were strongly affected by sampling location. The TL and n-3 PUFAs levels tended to be higher in temperate seaweeds compared with those in tropical seaweeds. The major n-3 PUFAs in different phyla, namely, eicosapentaenoic acid (EPA) and stearidonic acid (SDA) in Ochrophyta, alpha-linolenic acid (ALA) and SDA in Chlorophyta, and EPA in Rhodophyta, accumulated in temperate seaweeds. Chlorophylls, their derivatives, and carotenoids are known to have health benefits, such as antioxidant activities, while n-3 PUFAs are known to be essential nutrients that positively influence human nutrition and health. Therefore, seaweed lipids could be used as a source of ingredients with health benefits for functional foods and nutraceuticals.
• Carotenoid Profiling of a Red Seaweed Pyropia yezoensis: Insights into Biosynthetic Pathways in the Order Bangiales

  Jiro Koizumi, Naoki Takatani, Noritoki Kobayashi, Koji Mikami, Kazuo Miyashita, Yumiko Yamano, Akimori Wada, Takashi Maoka, Masashi Hosokawa

  MARINE DRUGS 16 (11) 1660-3397 2018/11 [Refereed][Not invited]
   

  Carotenoids are natural pigments that contribute to light harvesting and photo-protection in photosynthetic organisms. In this study, we analyzed the carotenoid profiles, including mono-hydroxy and epoxy-carotenoids, in the economically valuable red seaweed Pyropia yezoensis, to clarify the detailed biosynthetic and metabolic pathways in the order Bangiales. P. yezoensis contained lutein, zeaxanthin, -carotene, and -carotene, as major carotenoids in both the thallus and conchocelis stages. Monohydroxy intermediate carotenoids for the synthesis of lutein with an epsilon-ring from -carotene, -cryptoxanthin (,epsilon-caroten-3'-ol), and zeinoxanthin (,epsilon-caroten-3-ol) were identified. In addition, -cryptoxanthin, an intermediate in zeaxanthin synthesis from -carotene, was also detected. We also identified lutein-5,6-epoxide and antheraxanthin, which are metabolic products of epoxy conversion from lutein and zeaxanthin, respectively, by LC-MS and H-1-NMR. This is the first report of monohydroxy-carotenoids with an epsilon-ring and 5,6-epoxy-carotenoids in Bangiales. These results provide new insights into the biosynthetic and metabolic pathways of carotenoids in red seaweeds.
• Consumption of the edible sea urchin Mesocentrotus nudus attenuates body weight gain and hepatic lipid accumulation in mice

  Ryoko Yamamoto, Hisanori Minami, Hiromi Matsusaki, Mami Sakashita, Naoki Morita, Osamu Nishimiya, Naonobu Tsutsumi, Masashi Hosokawa, Yutaka Itabashi, Toshiro Matsui, Kazuhiro Urad

  JOURNAL OF FUNCTIONAL FOODS 47 40 - 47 1756-4646 2018/08 [Refereed]
   

  The beneficial health effects of edible sea urchin consumption in mice fed a normal (ND) or high-fat diet (HFD) were investigated in this study. Notably, sea urchin-administered (250, 500, 1000 mg/kg) mice exhibited lower body, liver, and visceral fat weights, lower plasma levels of aspartate aminotransferase and alanine aminotransferase, and lower hepatic triacylglycerol levels than those fed carboxymethyl cellulose (CMC). Despite the high levels of cholesterol found in sea urchins, intake of these organisms had no effect on plasma cholesterol levels among the mice tested. Additionally, sea urchin consumption resulted in enhanced levels of arachidonic acid and docosahexaenoic acid within mouse livers. Lastly, mice fed HFD with sea urchin (500 mg/kg) exhibited increased mRNA expression of uncoupling protein-1 within brown adipose tissue, compared with those fed HFD with CMC. In conclusion, consumption of sea urchin might provide a protective effect against the development of obesity and/or nonalcoholic fatty liver disease.
• Detection and identification of furan fatty acids from fish lipids by high-performance liquid chromatography coupled to electrospray ionization quadrupole time-of-flight mass spectrometry

  Hajime Uchida, Yutaka Itabashi, Ryuichi Watanabe, Ryoji Matsushima, Hiroshi Oikawa, Toshiyuki Suzuki, Masashi Hosokawa, Naonobu Tsutsumi, Kazuhiro Ura, Donato Romanazzi, Matthew R. Miller

  FOOD CHEMISTRY 252 84 - 91 0308-8146 2018/06 [Refereed][Not invited]
   

  Using high-performance liquid chromatography coupled to high-resolution electrospray ionization quadrupole time-of-flight mass spectrometry (HPLC/ESI-Q-TOF-MS), we have developed a new method for detection and identification of furan fatty acids (F-acids), which are widely distributed in living organisms and foods as minor lipid components and are known to have antioxidant and anti-inflammatory effects. For this purpose, total fatty acids prepared from the testis lipids of Japanese chum salmon (Oncorhynchus keta) were examined without any concentration or isolation of F-acids. In negative ESI mode, F-acids gave a prominent [M-H]-ion, by which individual F-acids could be detected and identified. High-resolution extracted ion chromatograms clearly showed the occurrence of five major F-acid homologs as already reported by GC/MS. The method was successfully applied to several fish samples and revealed the occurrence of F-acids for the first time in the two New Zealand fish, hoki (Macruronus novaezelandiae) and school shark (Galeorhinus galeus).
• Acrolein as a Major Volatile in the Early Stages of Fish Oil TAG Oxidation

  Ako Shibata, Mariko Uemura, Masashi Hosokawa, Kazuo Miyashita

  JOURNAL OF OLEO SCIENCE 67 (5) 515 - 524 1345-8957 2018/05 [Refereed][Not invited]
   

  Fish oil rich in docosahexaenoic acid (DHA) and eicosapentaenoic acid (EPA) is known to have an unpleasant smell, even at low oxidation levels. Therefore, it is highly important to know the major volatiles formed during the early stages of fish oil oxidation. Comparative study with solid-phase microextraction (SPME) and static headspace (SHS) methods showed that 2-propenal (acrolein) was formed as the major volatile from the beginning of fish oil triacylglycerol (TAG) oxidation. The effectiveness of SPME extraction on each volatile was different from each fiber. Among the three different SPME fibers used in the present study, carboxen/polydimethylsiloxane (CAR/PDMS) was determined to be a better fiber for measuring the volatiles, including acrolein. The present study also showed that the non-selective SHS method is useful for determining the characteristic volatile formation in the early stages of fish oil TAG oxidation.
• Comparison of oxidative stability of monogalactosyl diacylglycerol, digalactosyl diacylglycerol, and triacylglycerol containing polyunsaturated fatty acids.

  Hazahari, N.Y, B, Hosokawa, M, Miyashita, K

  Food Nutr. Sci. 9 221 - 234 2018 [Refereed][Not invited]
  
• Health impact of marine carotenoids.

  Miyashita, K, Hosokawa, M

  J. Food Bioact. 1:31-40 (2018) 1 34 - 40 2018 [Refereed][Not invited]
  
• 高度不飽和脂肪酸に対するスフィンゴイド塩基とα-トコフェロールの相乗的抗酸化効果

  宮下和夫, 上村麻梨子, 細川雅史

  オレオサイエンス 18 317 - 324 2018 [Refereed][Not invited]
  
• Effective Prevention of Oxidative Deterioration of Fish Oil: Focus on Flavor Deterioration

  Kazuo Miyashita, Mariko Uemura, Masashi Hosokawa

  ANNUAL REVIEW OF FOOD SCIENCE AND TECHNOLOGY, VOL 9 9 209 - 226 1941-1413 2018 [Refereed][Not invited]
   

  Docosahexaenoic acid (DHA) and eicosapentaenoic acid (EPA), both abundant in fish oil, are known to have significant biochemical and physiological effects primarily linked to the improvement of human health, especially cardiovascular and brain health. However, the incorporation of fish oil into foods and beverages is often challenging, as fish oil is very easily oxidized and can cause undesirable flavors. This review discusses this rapid formation of the fishy and metallic off-flavors, focusing especially on an early stage of fish oil oxidation. Although oxidative stability and quality of commercialized fish oil have improved over the past few years, there is a still a problem with its application: Flavor deterioration can be found even at very low oxidation levels. This review also notes the effective way to inhibit the formation of the volatile compounds responsible for the flavor deterioration.
• Spatial and seasonal variations in the biofunctional lipid substances (fucoxanthin and fucosterol) of the laboratory-grown edible Japanese seaweed (Sargassum horneri Turner) cultured in the open sea

  Masaru Terasaki, Chikara Kawagoe, Atsushi Ito, Hiroko Kumon, Bhaskar Narayan, Masashi Hosokawa, Kazuo Miyashita

  SAUDI JOURNAL OF BIOLOGICAL SCIENCES 24 (7) 1475 - 1482 1319-562X 2017/11 [Refereed][Not invited]
   

  This work studied the effect of spatial and seasonal differences on the accumulation of functional lipid components in Sargassum horneri (Turner), an edible Japanese seaweed popularly called Akamoku. S. horneri obtained from Samenoura bay area of Japan was laboratory cultured to evaluate the effect of temperature on the accumulation of total lipids (TL), fucoxanthin (Fx) and fucosterol (Fs) by the alga. The laboratory cultured 3 month old S. horneri were cultured in the open sea in two different geographical locations off Usujiri and Matsushima to evaluate the monthly variations, over a year, in their TL, Fx and Fs contents. S. horneri grown off the Usujiri area accumulated the maximum TL close to 193 mg g(-1) dry weight during the coldest part of the year. Fx and Fs contributed 5.6% and 16.2% of the TL in S. horneri harvested off Usujiri in February. Further, in spite of being the same species and parent stock, S. horneri grown off the Matsushima area accumulated less TL, Fx and Fs as compared to their Usujiri counterparts. Our study clearly indicates the role of temperature and light apart from nutritional profile and depth of waters where the seaweed was grown on the accumulation of functional lipid components in S. horneri. (C) 2016 The Authors. Production and hosting by Elsevier B.V. on behalf of King Saud University.
• Docosapentaenoic Acid (22:5n-3) Downregulates mRNA Expression of Pro-inflammatory Factors in LPS-activated Murine Macrophage Like RAW264.7 Cells

  Yanzhu Tian, Ami Katsuki, Donato Romanazzi, Matthew R. Miller, Serean L. Adams, Kazuo Miyashita, Masashi Hosokawa

  JOURNAL OF OLEO SCIENCE 66 (10) 1149 - 1156 1345-8957 2017/10 [Refereed][Not invited]
   

  Docosapentaenoic acid (22:5n-3, n-3 DPA) is a n-3 polyunsaturated fatty acid (PUFA) found in fish oil, and has been reported to have health benefits. This study investigated conversion of n-3 DPA, and examined the anti-inflammatory effects of n-3 DPA on activated macrophages. Murine macrophage-like RAW264.7 cells were incubated in culture media containing n-3 DPA for 72 h. The level of n-3 DPA in the fatty acid composition of the total lipid fraction increased in a dose-dependent manner. Furthermore, the levels of docosahexaenoic acid (DHA) and eicosapentaenoic acid (EPA) were higher in treated cells than in control cells. In RAW264.7 cells stimulated by lipopolysaccharide (LPS), n-3 DPA significantly down regulated mRNA expression of pro-inflammatory factors such as IL-6, IL-1 beta, iNOS and COX-2. Production of IL-6 was also reduced by n-3 DPA in a dose-dependent manner. We found that n-3 DPA treatment resulted in greater IL-6 mRNA down-regulation than that achieved with EPA treatment, and was similar to that of DHA treatment. Furthermore, expression levels of IL-6 and IL-1 beta mRNAs were measured in the presence of the delta-6 desaturase inhibitor SC26196 in the culture medium to inhibit the conversion of n-3 DPA to DHA. There was no significant difference in the down-regulation in the mRNA expression of pro inflammatory cytokines in RAW264.7 cells by n-3 DPA with or without presence of SC26196. These results demonstrate that n-3 DPA exhibits anti-inflammatory effects in activated RAW264.7 cells, which are independent of DHA conversion.
• Tetrahydrobiopterin activates brown adipose tissue and regulates systemic energy metabolism.

  Oguri Y, Fujita Y, Abudukadier A, Ohashi A, Goto T, Furuya F, Obara A, Fukushima T, Matsuo N, Kim M, Hosokawa M, Kawada T, Hasegawa H, Inagaki N

  JCI Insight 9 91981  2017/05 [Refereed][Not invited]
  
• Fatty Acid and Lipid Class Composition of the Microalga Phaeodactylum tricornutum

  Yu-Hong Yang, Lei Du, Masashi Hosokawa, Kazuo Miyashita, Yume Kokubun, Hisayoshi Arai, Hiroyuki Taroda

  JOURNAL OF OLEO SCIENCE 66 (4) 363 - 368 1345-8957 2017/04 [Refereed][Not invited]
   

  n-3 Polyunsaturated fatty acids (PUFAs) are essential for the prevention and/or risk reduction of some diseases, including cardiovascular diseases and cancer. Therefore, PUFAs-rich marine microalgae have received considerable research attention. The diatom Phaeodactylum tricornutum, a rich source of lipids and fucoxanthin, has commercial applications in the food and pharmaceutical industries. In this study, the lipid class composition and fatty acid distribution in P. tricornutum under high-density and scaleup cultivation were investigated by thin-layer chromatography and gas chromatography. The fucoxanthin content was quantified by reverse-phase high-performance liquid chromatography. Both total lipid and fucoxanthin contents were high, accounting for 321.89 and 4.47 mg/g dry weight, respectively. Neutral lipids were the major lipids, and triacylglycerol was the predominant neutral lipids (33.63% of total lipids). The glycolipids (GLs) and phospholipids (PLs) represented 20.95% and 31.39% of total lipids, respectively. Sulfoquinovosyl diacylglycerol (SQDG) was the largest GLs fraction (9.81% of total lipids). Phosphatidylglycerol (PG) was the major PLs and accounted for 13.53% of total lipids. The main fatty acids were hexadecanoic acid (C16:0), palmitoleic acid (C16:1), and eicosapentaenoic acid. In addition, docosahexaenoic acid only accumulated in the PLs fraction. These findings provide new information on the lipid types, fatty acid composition of each lipid class, and the fucoxanthin content in P tricornutum.
• Reduction of HbA1c levels by fucoxanthin-enriched akamoku oil possibly involves the thrifty allele of uncoupling protein 1 (UCP1): a randomised controlled trial in normal-weight and obese Japanese adults

  Nana Mikami, Masashi Hosokawa, Kazuo Miyashita, Hitoshi Sohma, Yoichi M. Ito, Yasuo Kokai

  JOURNAL OF NUTRITIONAL SCIENCE 6 e5  2048-6790 2017/02 [Refereed][Not invited]
   

  Lifestyle-related problems are becoming a major health threat in East Asian countries. Therefore, finding an efficacious nutraceutical for this population is important. One candidate is fucoxanthin (Fx), a carotenoid abundantly found in edible brown seaweed that has been associated with a number of valuable health-promoting benefits. Unfortunately, clinical studies of Fx are limited. In the present study, we aimed to evaluate the effects of Fx on obesity-related parameters in Japanese subjects harbouring an SNP associated with lifestyle-related problems. In all, sixty normal-weight and obese Japanese adults with BMI over 22 kg/m(2) were single-blinded and randomly assigned to three Fx-dose cohorts and administered Fx-enriched akamoku oil containing Fx at 0, 1 or 2 mg/d for 8 weeks (n 20 per group). Parameters relating to obesity and serum Fx metabolites were measured before and after intervention, but no significant differences were observed between and within the groups. Despite no changes in visceral fat areas and resting energy expenditures after intervention, we observed a significant decline in HbA1c levels in the 2 mg/d Fx group compared with that in the 0 mg/d group (P < 0.05), which was correlated with an increase in serum fucoxanthinol (Fx metabolite) levels. In addition, HbA1c levels declined more significantly in subjects with G/G alleles of the uncoupling protein 1 (UCP1) gene than in those with the A/A and A/G alleles (P < 0.05). We conclude that although Fx supplementation does not affect visceral fat areas, it may reduce HbA1c levels in those harbouring the thrifty allele of UCP1-3826A/G.
• Squalene modulates fatty acid metabolism: Enhanced EPA/DHA in obese/diabetic mice (KK-A(y)) model

  Sangeetha Ravi Kumar, Ippei Yamauchi, Bhaskar Narayan, Ami Katsuki, Masashi Hosokawa, Kazuo Miyashita

  EUROPEAN JOURNAL OF LIPID SCIENCE AND TECHNOLOGY 118 (12) 1935 - 1941 1438-7697 2016/12 [Refereed][Not invited]
   

  Biosynthesis of long-chain n-3 fatty acids from precursors is limited. In vivo effect of squalene (SQ) on the metabolic fate of n-3 fatty acid precursors in obese/diabetic KK-A(y) rodent model was evaluated in our work. Soybean oil, being rich in ALA (18:3 n-3; a known precursor of EPA/DHA), was chosen as the n-3 fatty acid precursor rich source. A high-fat diet (20%) containing 7% soybean oil (SO) was fed to obesity/diabetes-prone male KK-A(y) mice (control). In the case of diets fed to test groups, soybean oil was replaced with 5% SO and 2% SQ. Hepatic DHA levels increased (four fold) in SQ fed group over control (p<0.05). Gene and protein expressions of (5) and (6) desaturases, key enzymes involved in the fatty acid metabolism, further supported the results. Also, SQ exhibited a hypotriglyceridemic and hypoglycemic effect. The results clearly indicated the effect of SQ in modulating the n-3 fatty acid metabolism, including EPA/DHA synthesis in the presence of n-3 fatty acid precursor. This is the first report of enhancement of in vivo DHA/EPA by SQ and in turn, modulating the physiological fatty acid profile. Practical applications: Squalene (SQ) is an important marine biofunctional material that is found in some terrestrial sources as well. Squalene, being a cholesterol precursor, forms an interesting subject of research for its effect in vivo. SQ significantly enhanced proportions of EPA and/or DHA when their n-3 fatty acid precursors were available in the diet. The study further establishes the usefulness of SQ in functional food formulations. The work provides an important basis for further evaluation of the role of SQ in normal and disease conditions.KK-A(y) mice were fed high fat/sucrose diet to induce obesity/diabetes. Fat source in control diet was lard and soybean oil while experimental group diet contained 2% squalene+13% lard+5% soybean oil. Feeding squalene for 4weeks modulated fatty acid metabolism with increased docosahexaenoic acid (DHA) and decrease in triglycerides (TG), compared to control. The enhanced DHA in the fatty acid profile was supported by upregulated mRNA expression of (5)-desaturase enzyme and protein expression of (5) and (6) desaturases (FADS1 and FADS2). Additionally, squalene had a hypoglycemic effect in the mice.
• Bioconversion of Docosapentaenoic Acid in Human Cell Lines, Caco-2, HepG2, and THP-1

  Yanzhu Tian, Donato Romanazzi, Kazuo Miyashita, Masashi Hosokawa

  JOURNAL OF OLEO SCIENCE 65 (12) 1017 - 1022 1345-8957 2016/12 [Refereed][Not invited]
   

  Docosapentaenoic acids (DPAs) are long chain polyunsaturated fatty acids that exist as two major structural isomers: n-3 DPA and n-6 DPA. n-3 DPA is found in seal meat, salmon and abalone, and n-6 DPA is found in several marine microbial oil. We investigated the bioconversion of n-3 and n-6 DPAs in three different human cell lines, Caco-2, HepG2, and THP-1. n-3 DPA was converted to docosahexaenoic acid only in HepG2 cells. In contrast, retro-conversion to eicosapentaenoic acid (EPA) was observed in all three cell lines. n-6 DPA was also retro-converted to arachidonic acid (AA) in Caco-2 and HepG2 cells. EPA and AA were particularly elevated in Caco-2 cells, compared to HepG2 cells. Further, the retro-conversion of n-3 DPA led to a greater increase of EPA in the phospholipid fraction than in the neutral lipid fraction.
• Inhibitory Effect of Dihydrosphingosine with alpha-Tocopherol on Volatile Formation during the Autoxidation of Polyunsaturated Triacylglycerols

  Mariko Uemura, Ako Shibata, Masashi Hosokawa, Ai Iwashima-Suzuki, Makoto Shiota, Kazuo Miyashita

  JOURNAL OF OLEO SCIENCE 65 (9) 713 - 722 1345-8957 2016/09 [Refereed][Not invited]
   

  The effect of dihydrosphingosine (d18:0) on triacylglycerol (TAG) oxidation was examined with and without alpha-tocopherol. Three types of TAG from fish, linseed, and soybean oil were oxidized at 50 degrees C to determine the effect of dihydrosphingosine (d18:0) with or without alpha-tocopherol on triacylglycerol (TAG) oxidation. The analysis of oxygen consumption and total volatile formation demonstrated a small effect of d18:0 on TAG oxidation in the absence of alpha-tocopherol. On the other hand, the combination of d18:0 with alpha-tocopherol showed strong antioxidant activity and completely inhibited volatile formation within 1400 h for soybean oil TAG, 650 h for linseed oil TAG, and 380 h for fish oil TAG.
• Combined effect of astaxanthin and squalene on oxidative stress in vivo

  Sangeetha Ravi Kumar, Bhaskar Narayan, Yuki Sawada, Masashi Hosokawa, Kazuo Miyashita

  MOLECULAR AND CELLULAR BIOCHEMISTRY 417 (1-2) 57 - 65 0300-8177 2016/06 [Refereed][Not invited]
   

  Obesity and diabetes, risk factors for metabolic syndrome, are characterized by oxidative stress and inflammatory responses. Marine biofunctionals, astaxanthin (Ax) and squalene (SQ), were evaluated for their combined effect. Groups of male KK-A (y) mice were fed high fat/sucrose diet for 4 weeks, supplemented with either 0.1 %Ax, 2 %SQ or 0.1 %Ax + 2 %SQ. In comparison to control, Sod was elevated in only Ax + SQ. However, Gpx was highest in Ax + SQ, indicating the combined antioxidant effect of Ax and SQ. This was supported by elevated mRNA expression of Sod1 and Gpx1. Except adiponectin (elevated in Ax and Ax + SQ), expression of other inflammatory markers was not altered. Blood glucose levels were decreased in SQ and Ax + SQ while liver triglycerides decreased in SQ group. This is the first in vivo study demonstrating combined effects of Ax and SQ resulting in antioxidant effects and modulation of glucose/triglyceride levels. This study highlights the benefit of utilizing Ax and SQ together for management of obesity/diabetes.
• Does squalene alter the antioxidant potential of astaxanthin and fucoxanthinol? In vitro evidence in RAW 264.7 cells, a murine macrophage

  Sangeetha Ravi Kumar, Bhaskar Narayan, Yuki Kizawa, Masashi Hosokawa, Kazuo Miyashita

  JOURNAL OF FOOD SCIENCE AND TECHNOLOGY-MYSORE 53 (4) 2139 - 2143 0022-1155 2016/04 [Refereed][Not invited]
   

  Astaxanthin (Ax) and fucoxanthin/fucoxanthinol (FuOH) are marine xanthophylls exhibiting anti-oxidant effects. Squalene (SQ) is a triterpenoid and is a precursor of sterols. This study aimed to determine if SQ can improve the effect of Ax/FuOH on lipid peroxidation. RAW 264.7 cells were treated with different concentrations of Ax, FuOH and SQ and corresponding rate of cell survival was noted. In addition,combination groups - Ax + SQ and FuOH + SQ- were also run. Cells treated with Ax, FuOH, SQ, Ax + SQ and FuOH + SQ were stimulated with lipopolysaccharide and lipid hydroperoxides were estimated. Results showed that 5 mu M Ax, 2 mu M FuOH and 10 mu M SQ supported cell survival. In presence of SQ, cell viability improved for higher concentrations of FuOH (5, 10 mu M). Lipid hydroperoxides were supressed by Ax, FuOH, Ax + SQ and FUOH +SQ and were significantly lower in Ax + SQ, indicating the synergistic effect of Ax and SQ. To conclude, combination of Ax with SQ enhances its ability to supress lipid peroxidation while with FuOH, SQ attenuates the toxic effect at higher doses. Moreover, this is the first time that the combined effect of SQ and carotenoids has been studied and reported.
• Lipids, Fatty Acids, and Fucoxanthin Content from Temperate and Tropical Brown Seaweeds

  Eko Susanto, Akhmad Suhaeli Fahmi, Masayuki Abe, Masashi Hosokawa, Kazuo Miyashita

  2ND INTERNATIONAL SYMPOSIUM ON AQUATIC PRODUCTS PROCESSING AND HEALTH, ISAPPROSH 2015 7 66 - 75 2214-241X 2016 [Refereed][Not invited]
   

  Brown seaweeds lipid fraction contains several bioactive components such as Fx, polyphenol and n-3 PUFA. In this research, total lipids, Fx and FA compositions of brown seaweeds harvested from cold waters and warm waters were evaluated. The seven brown seaweeds studied were collected in different months from two different geographical areas, viz. cold waters (Japan) and warm waters (Indonesia). The result show that total lipid and Fucoxanthin in temperate brown seaweeds were higher than tropical brown seaweeds. The major PUFA from warm water seaweeds were 16: 0, 18: 1n-9, 20: 4n-6, and cold water continued dominantly 16: 0, 20: 4n-6, 20: 5n-3. Temperate brown seaweeds [S. horneri (Turner) J. Agardh] was rich in fucoxanthin and n-3 PUFA especially EPA. (C) 2016 The Authors. Published by Elsevier B.V.
• A protocol for human serum fucoxanthinol quantitation using LC-MS/MS system.

  Mikami, N, Hosokawa, M, Abe, M, Miyashita, K, Sohma, H, Kokai, Y

  J. Nutri. Med. Diet. Care. 2 019  2016 [Refereed][Not invited]
  
• The dietary effect of milk sphingomyelin on the lipid metabolism of obese/diabetic KK-A(y) mice and wild-type C57BL/6J mice

  Ippei Yamauchi, Mariko Uemura, Masashi Hosokawa, Ai Iwashima-Suzuki, Makoto Shiota, Kazuo Miyashita

  FOOD & FUNCTION 7 (9) 3854 - 3867 2042-6496 2016 [Refereed][Not invited]
   

  Purified milk sphingomyelin (SM) was obtained from lipid concentrated butter serum (LC-BS) by successive separations involving solvent fractionation, selective saponification, and silicic acid column chromatography. The SM obtained was given to obese/diabetic KK-A(y) mice and wild-type C57BL/6J mice. SM supplementation significantly increased fecal lipids paralleled with a decrease in non-HDL cholesterol levels in the serum and neutral lipids and in cholesterol levels in the livers of KK-A(y) mice. The reduction of liver lipid levels also resulted in a decrease in the total fatty acid content of the KK-A(y) mice livers, while n-3 fatty acids derived from the conversion of a-linolenic acid (18:3n-3) increased due to SM supplementation. In contrast to the KK-A(y) mice, little change in the serum and liver lipids was observed in wild-type C57BL/6J mice. The present study suggests that SM may be effective only in subjects with metabolic disorders.
• Dietary ALA from Spinach Enhances Liver n-3 Fatty Acid Content to Greater Extent than Linseed Oil in Mice Fed Equivalent Amounts of ALA

  Miho Kuroe, Hiroyuki Kamogawa, Masashi Hosokawa, Kazuo Miyashita

  LIPIDS 51 (1) 39 - 48 0024-4201 2016/01 [Refereed][Not invited]
   

  Although several works have reported absorption rate differences of n-3 polyunsaturated fatty acids (PUFA) bound to different lipid forms, such as ethyl ester, triacylglycerol (TAG), and phospholipids, no studies have investigated the effect of n-3 PUFA from glycolipids (GL). The present study compared the fatty acid contents of tissue and serum lipids from normal C57BL/6J mice fed two types of alpha-linolenic acid (ALA)-rich lipids, spinach lipid (SPL), and linseed oil (LO). ALA was primarily present as the GL form in SPL, while it existed as TAG in LO. Supplementation of both lipids increased ALA and its n-3 metabolites, eicosapentaenoic acid (EPA), docosapentaenoic acid (DPA), and docosahexaenoic acid, and decreased n-6 PUFA, linoleic acid and arachidonic acid, in the livers, small intestines, and sera of the treated mice compared with those of the control group. When the comparison between the SPL and LO diets containing the same amount of ALA was conducted, the EPA and DPA levels in the liver lipids from mice fed the SPL diet were significantly higher than those fed the LO diet. Additionally, the total contents of n-3 PUFA of lipids from the livers, small intestines, and sera of the SPL group were higher than those of the LO group.
• Formation of Acrolein in the Autoxidation of Triacylglycerols with Different Fatty Acid Compositions

  Ako Shibata, Mariko Uemura, Masashi Hosokawa, Kazuo Miyashita

  JOURNAL OF THE AMERICAN OIL CHEMISTS SOCIETY 92 (11-12) 1661 - 1670 0003-021X 2015/12 [Refereed][Not invited]
   

  Fish, echium, linseed, and soybean oil triacyglycerols (TAGs) were oxidized at 50 or 60 A degrees C to determine the effect of the polyunsaturated fatty acid composition on the volatile product formation. The analysis of the oxygen consumption and total volatile formation demonstrated that the soybean oil TAG had the highest oxidative stability followed by linseed, echium, and fish oil TAGs. Our results were in agreement with the expected average number of bis-allylic positions of each TAG. Higher quantities of acrolein (2-propenal) and propanal were detected using the static headspace gas chromatography method at the early stages of oxidation of echium and fish oil TAGs; however, a considerable amount of propanal and only a small amount of acrolein were found in the oxidized linseed oil TAG. The peak area ratios of acrolein to propanal were 0.115, 0.569, and 2.554 after the 8-h oxidation of linseed, echium, and fish oil TAG, respectively, suggesting the preferential formation of acrolein, especially during the fish oil TAG oxidation. The acrolein quickly increased during the first stage of oxidation, but thereafter it either did not change or slightly decreased during the fish oil oxidation. Because fish oil induces flavor deterioration from the very early stage of the oxidation, the acrolein formation observed in the present study may be important for fish oil deterioration.
• The Beneficial Health Effects of Fucoxanthin

  Kazuo Miyashita, Masashi Hosokawa

  Genomics, Proteomics and Metabolomics in Nutraceuticals and Functional Foods: Second Edition 122 - 134 2015/10/05 [Not refereed][Not invited]
   

  The beneficial effects of carotenoids on human health are thought to come from their antioxidant properties. However, there are other physiological activities that carotenoids have that may play a larger role in influencing human health. The beneficial effect of fucoxanthin supplementation on improving metabolic disorders such as hyperglycemia is due largely to its regulatory effect on gene and protein expression rather than the ability of fucoxanthin to scavenge active oxygen species and/or free radicals. Fucoxanthin inhibits cancer cell growth more effectively than other carotenoids by arresting the cell cycle and triggering apoptosis. The higher activity of fucoxanthin is related to the allenic bond of its chemical structure. This unique structure is responsible for the anti-obesity effect based on the specific molecular mechanism that has not been found in other carotenoids. Nutrigenomic studies reveal that fucoxanthin induces uncoupling protein 1 (UCP1) expression in abdominal white adipose tissue (WAT) mitochondria that leads to the oxidation of fatty acids and heat production in the WAT. The induction of UCP1 expression in abdominal WAT is unusual because UCP1 is usually only found in brown adipose tissue. Fucoxanthin supplementation also improves insulin resistance and decreases blood glucose levels through the regulation of adipokine secretions from WAT and the upregulation of glucose transporter 4 expression and its translocation to muscle cell membranes.
• Anticancer effects of fucoxanthin and fucoxanthinol on colorectal cancer cell lines and colorectal cancer tissues

  Kazuto Takahashi, Masashi Hosokawa, Hiroyuki Kasajima, Kazuteru Hatanaka, Kazuhiro Kudo, Norihiko Shimoyama, Kazuo Miyashita

  ONCOLOGY LETTERS 10 (3) 1463 - 1467 1792-1074 2015/09 [Refereed][Not invited]
   

  Colorectal cancer is one of the most malignant neoplasms worldwide. Fucoxanthin is a carotenoid present in the chloroplasts of brown seaweeds. In the present study, the anticancer effects of fucoxanthin and its metabolite, fucoxanthinol, on 6 colorectal cancer cell lines and 20 tissue samples from surgically resected Clinical colorectal cancer specimens were examined using a collagen-gel droplet embedded culture drug sensitivity test (CD-DST). The in vitro sensitivity to fiicoxanthin, fucoxanthinol and the anticancer drugs is expressed as T/C (%), where T is the absorbance of cells which stained by neutral red treated with carotenoids and C is the absorbance of non-staining cells. Fucoxanthin and fucoxanthinol decreased the T/C (%) of Caco-2, WiDr, HCI116, and DLD-1 cell lines at doses of 20 mu M. Fucoxanthinol also decreased the TIC (%) of SW620 cells, while the T/C (%) of Colo205 cells was not reduced by treatment with either carotenoid. Specifically, the TIC (%) of Caco-2 and Wi Dr cells, which were incubated in carotenoid-free medium for 6 days following treatment with 20 mu M fucaxanthinol for 24 h, was markedly decreased to 1.4 +/- 0.2 and 12.0 +/- 0.3%, respectively. Furthermore, fucoxanthin and fucoxanthinol decreased the TV (%) in colorectal cancer tissue samples. Notably, 20 mu M fucoxanthinol treatment resulted in a higher proportion of colorectal cancer samples with a T/C (%) of <50% (13/20, 65%) compared with samples treated with 20 mu M fucoxanthin (2/20, 10%). The median TIC (%) value of 35.1% for the 20 cancers specimens treated with 20 mu M fucoxanthinol was lower than the median T/C (%) values of 86.3% and 75.8% for those treated with fluorouracil and paclitaxel, respectively. These results suggested that fucoxanthin and fucoxanthinol may be of use as Chemotherapeutic agents in colorectal cancer.
• Fucoxanthinol, Metabolite of Fucoxanthin, Improves Obesity-Induced Inflammation in Adipocyte Cells

  Hayato Maeda, Shogo Kanno, Mei Kodate, Masashi Hosokawa, Kazuo Miyashita

  MARINE DRUGS 13 (8) 4799 - 4813 1660-3397 2015/08 [Refereed][Not invited]
   

  Fucoxanthin (Fx) is a marine carotenoid found in edible brown seaweeds. We previously reported that dietary Fx metabolite into fucoxanthinol (FxOH), attenuates the weight gain of white adipose tissue of diabetic/obese KK-A(y) mice. In this study, to evaluate anti-diabetic effects of Fx, we investigated improving the effect of insulin resistance on the diabetic model of KK-A(y) mice. Furthermore, preventing the effect of FxOH on low-grade chronic inflammation related to oxidative stress was evaluated on 3T3-L1 adipocyte cells and a RAW264.7 macrophage cell co-culture system. A diet containing 0.1% Fx was fed to diabetic model KK-A(y) mice for three weeks, then glucose tolerance was observed. Fx diet significantly improved glucose tolerance compared with the control diet group. In in vitro studies, FxOH showed suppressed tumor necrosis factor- (TNF-), and monocyte chemotactic protein-1 (MCP-1) mRNA expression and protein levels in a co-culture of adipocyte and macrophage cells. These findings suggest that Fx ameliorates glucose tolerance in the diabetic model mice. Furthermore, FxOH, a metabolite of Fx, suppresses low-grade chronic inflammation in adipocyte cells.
• Structure of a novel monocyclic carotenoid, 3 ''-hydroxy-2 '-isopentenylsaproxanthin ((3R,2 ' S)-2 -(3-hydroxy-3 -methylbuty1)-3 ', 4 '-didehydro-1 ', 2 '-dihydro-beta, Psi-carotene-3, 1 '-diol), from a flavobacterium Gillisia limnaea strain DSM 15749

  Naoki Takatani, Tomoo Sawabe, Takashi Maoka, Kazuo Miyashita, Masashi Hosokawa

  BIOCATALYSIS AND AGRICULTURAL BIOTECHNOLOGY 4 (2) 174 - 179 1878-8181 2015/04 [Refereed][Not invited]
   

  Gillisia lirtinaea strain DSM 15749 (=R-8282(T), =LMG21470,=CIP108418) is a Gram-negative, strictly aerobic, psychrophilic, and non-photosynthetic bacterium that belongs to the family Flavobacteriaceae, which was isolated from a microbial mat in Lake Fryxell, Antarctica (Van. Trapp en eI. al., 2004). The bacteriurn shows yellow colored colony, but the cat otenoid composition has never been determined yet. In this study, we elucidated the structure of carotenoids produced by the strain DSM 15749. G. lirtinaea strain DSM 15749 synthesized a novel monocyclic-type carotenoid, 3 ''-hydroxy-2'-isopentenylsaproxanthin ((3R, 2'S')-2 '-(3-hydroxy-3-methylbutyl)-3 '', 4'-didehydro-1', 2'-dihydro-beta, Psi-carotene-3, 1'-diol), as well as (3R, TR)-zeaxanthin. Furthermore, another monocyclic carotenoid, 2'-isopentenylsaproxanthin, was also detected in the strain DSM 15749. 3 ''-Hydroxy-2'-isopentenylsaproxanthin is "Chimera'like unique structures, in that one end group forms the same structure of zeaxanthin, and the other end group is the same as bacterioruberin which is produced by several extremophilic bacteria and archaea. This structure involved the hydroxy derivative of 2'-isopentenylsaproxanthin. The present study is the First report about the carotenoids produced by the genus Gillisto, of the family Flavobacteriaceae. (C) 2015 Elsevier Ltd. All rights reserved.
• 褐藻カロテノイド、フコキサンチン

  宮下和夫, 細川雅史

  FFIジャーナル FFIジャーナル編集委員会 220 (2) 101 - 109 0919-9772 2015 [Not refereed][Not invited]
  
• Identification of a novel carotenoid, 2 '-isopentenylsaproxanthin, by Jejuia pallidilutea strain 11shimoA1 and its increased production under alkaline condition

  N. Takatani, K. Nishida, T. Sawabe, T. Maoka, K. Miyashita, M. Hosokawa

  APPLIED MICROBIOLOGY AND BIOTECHNOLOGY 98 (15) 6633 - 6640 0175-7598 2014/08 [Refereed][Not invited]
   

  Carotenoids are a class of naturally occurring pigment, carrying out important biological functions in photosynthesis and involved in environmental responses including nutrition in organisms. Saproxanthin and myxol, which have monocyclic carotenoids with a gamma-carotene skeleton, have been reported to show a stronger antioxidant activity than those with beta-carotene and zeaxanthin. In this research, a yellow-orange bacterium of strain 11shimoA1 (JCM19538) was isolated from a seaweed collected at Nabeta Bay (Shizuoka, Japan). The 16S rRNA gene sequence of strain 11shimoA1 revealed more than 99.99 % similarity with those of Jejuia pallidilutea strains in the family Flavobacteriaceae. Strain 11shimoA1 synthesized two types of carotenoids. One of them was (3R, 3'R)-zeaxanthin with dicyclic structure and another was identified as (3R, 2'S)-2'-isopentenylsaproxanthin, a novel monocyclic carotenoid with pentenyl residue at C-2' position of saproxanthin, using FAB-MS, H-1 NMR, and CD analyses. Culturing strain 11shimoA1 in an alkaline medium at pH 9.2 resulted in a markedly increased in production of 2'-isopentenylsaproxanthin per dry cell weight, but a decreased in zeaxanthin production as compared to their respective production levels in medium with pH 7.0. These carotenoids are likely to play some roles in the adaptation of the bacterium to the environmental conditions.
• Source-sink manipulation of Camelina sativa L. related to grain yield under stressful environment of Hokkaido, Japan

  Kounosuke Fujita, Shohei Fujita, Takayuki Fujita, Satoshi Konishi, Johann Vollmann, Pravat Kumar Mohapatra, Masashi Hosokawa, Masato Kuwahara, Hirofumi Saneoka, Toshiaki Tadano

  SOIL SCIENCE AND PLANT NUTRITION 60 (2) 156 - 161 0038-0768 2014/04 [Refereed][Not invited]
   

  Five cultivars of camelina species (Camelina sativa L.) were grown in the field at two levels of nitrogen (N) under the climate conditions of Hokkaido Island, Japan. N nutrition as well as the genetic difference between cultivars influenced biomass and grain production. Three cultivars responded positively to N application, but not the other two. Increase in grain yield was associated more with grain number than grain size. The plants accommodated the extra grains by increasing pod number. Irrespective of the N level in the soil, a linolenic acid concentration of the grain was very high at 31% of the total fatty acids. In cultivar Calena, lower leaves on the main stem were excised to find the effect of reduced source area on yield. Defoliation of lower leaves did not affect grain yield, indicating that these leaves do not contribute to source capacity of the plant during grain filling. However, removal of all leaves decreased grain yield to 70% relative to the control plant. In contrast to defoliation, shading of the whole panicle consisting of the pod shell and rachis was more effective in reducing grain yield. Shading of the panicle prior to the grain filling period at the levels of 33 and 6% irradiance relative to control decreased grain yield to 39 and 26%, respectively. These results suggested that the pod shell and rachis are more important source organs for photosynthetic carbon supply to grains than the upper leaves, and they can sustain grain filling in exigency. The presence of attributes like extended source area for grain filling and high alpha-linolenic acid content of leaves is an indicator for successful adaptation of Camelina in the stress-prone environment of Hokkaido Island.
• The extracts of Japanese willow tree species are effective forapoptotic desperation or differentiation of acute myeloid leukemia cells

  Kounosuke Fujita, Yuji Nomura, Masahiko Sawajiri, Pravat K. Mohapatra, Hany A. El-Shemy, Nguyen T. Nguyen, Masashi Hosokawa, Kazuo Miyashita, Teruo Maeda, Hirofumi Saneoka, Shohei Fujita, Takayuki Fujita

  PHARMACOGNOSY MAGAZINE 10 (38) 125 - 131 0973-1296 2014/04 [Refereed][Not invited]
   

  Background: The antileukemic activity of hot water extract of plant parts of some Japanese willow tree species grown at different levels of nitrogen were examined. Materials and Methods: Water extracts of willow leaves were prepared for this studies in different level of nitrogen nutrition. Results: The extracts obtained from the leaves and stem exhibited anti-leukemic activities prominently. The crude hot water extracts of the young growing parts including apex, matured leaves and stem, killed the blasts of acute myeloid leukemia (AML) cells, (HL60 and NB4) after 48h incubation, however, such desperation was far less in the root extract. Similar to the plant parts, response of extracts obtained from different willow species was not identical; the proportion of dead cells relative to whole cells of the culture medium ranged from 21% to 93% among the species. Leaf extracts obtained from the responsive willow species decreased the live cell percentage and increased the dead cell percentage at higher level of nitrogen nutrition. The mode of desperation of leaf extract treated AML cells in such species appeared to be cell apoptosis as shown by binding with fluorescein isothiocyanate (FITC) -labeled Annexin V. Conclusion: Differentiation of alive AML cells continued unabated and apoptosis was poor when extract of an unresponsive species added to the culture medium.
• Regulation of Polyunsaturated Fatty Acid Biosynthesis by Seaweed Fucoxanthin and Its Metabolite in Cultured Hepatocytes

  Tsunehiro Aki, Masaya Yamamoto, Toshiaki Takahashi, Kohki Tomita, Rieko Toyoura, Kazuhiro Iwashita, Seiji Kawamoto, Masashi Hosokawa, Kazuo Miyashita, Kazuhisa Ono

  LIPIDS 49 (2) 133 - 141 0024-4201 2014/02 [Refereed][Not invited]
   

  The effects of a seaweed carotenoid, fucoxanthin, and its physiological metabolite, fucoxanthinol, on the biosynthesis of polyunsaturated fatty acids (PUFA) were investigated using cultured rat hepatoma BRL-3A. The metabolism of alpha-linolenic acid (18:3n-3) was suppressed by the addition of these carotenoids, resulting in a decrease in the content of eicosapentaenoic acid (20:5n-3), which suggested a down-regulation of metabolic enzymes such as fatty acid desaturase and elongase. An increase in the content of docosahexaenoic acid (22:6n-3), as observed in previous studies in vivo, might be a buffering action to maintain the membrane fluidity. The suppressive effect of fucoxanthinol on a dagger 6 fatty acid desaturase was not at the level of gene expression but due to specific modifications of the protein via a ubiquitin-proteasome system. A proteomic analysis revealed several factors such as phosphatidylethanolamine-binding protein that might be involved in the observed action of fucoxanthin. These findings will contribute to studies on the elucidation of the precise molecular mechanisms underlying the regulation of PUFA biosynthesis by fucoxanthin.
• Carotenoids as a Nutraceutical Therapy for Visceral Obesity

  Kazuo Miyashita, Masashi Hosokawa

  Nutrition in the Prevention and Treatment of Abdominal Obesity 329 - 340 2014 [Refereed][Not invited]
   

  Obesity is linked to several diseases with a rapidly growing prevalence in most of the world. A few molecular targets are key to the successful development of antiobesity therapeutics: one of the most promising targets for activation is uncoupling protein 1 (UCP 1). Fucoxanthin, a marine carotenoid found in edible brown seaweeds, shows antiobesity effect that are mainly based on increasing energy expenditure in abdominal white adipose tissue (WAT) through UCP 1 induction. The key fucoxanthin structure involved in the antiobesity effect is suggested to be carotenoid end of the polyene chromophore containing an allenic bond. Fucoxanthin also improves insulin resistance and decreases blood glucose level through regulating adipocytokine secretions from abdominal WAT. Another important molecular mechanism for improving glucose utilization by fucoxanthin is solute carrier family 2, facilitated glucose transporter member 4 (GLUT4) upregulation and its translocation to the cell membrane from the cytosol. Interestingly, fucoxanthin effects are seen only in subjects with obesity and metabolic disorders, and not in normal healthy individuals. Therefore, fucoxanthin is a promising nutraceutical drug for treating obesity and related metabolic disorders. © 2014 Elsevier Inc. All rights reserved.
• Studies on chemical composition of three species of Enteromorpha

  K. Ganesan, K. Suresh Kumar, P. V. Subba Rao, Y. Tsukui, N. Bhaskar, M. Hosokawa, K. Miyashita

  Biomedicine and Preventive Nutrition 4 (3) 365 - 369 2210-5239 2014 [Refereed][Not invited]
   

  Studies on proximate, mineral, fatty and amino acid composition of edible Enteromorpha species, viz. Enteromorphacompressa, Enteromorphalinza and Enteromorphatubulosa, collected from the Northwest coast of India, revealed for species-dependent nutrient composition variation. The present study showed a higher amount of sugar (51.05. ±. 1.22%), protein (19.09. ±. 0.91%) and lipid content (5.56. ±. 0.16%) E.tubulosa. Amongst the seventeen elements analyzed (Ca, K, Mg, Na, P, As, Cd, Co, Cu, Fe, Hg, Mn, Mo, Ni, Zn, Cr, Pb), all three species showed a higher amount of calcium content. E.compressa was obtained with rich amount of macro-elements (11.42. ±. 0.017. g/100. g dry wt), while maximum micro- and trace-elements (81.51. ±. 6.65. mg/100. g dry wt) were recorded in E.linza. Amino acid analysis revealed seven out of the 10 essential amino acids were present either in excess or at-par with the reference proteins. Moreover, all the three species studied possessed relatively high n-3 fatty acids additionally they had more unsaturated fatty acids than saturate ones, except in E.compressa. Therefore, based on the present investigation, it could be stated that the three species studied could be used as food supplements to improve the nutritive value in the omnivorous diet. © 2014 Elsevier Masson SAS.
• Draft genome sequences of marine flavobacterium Nonlabens strains NR17, NR24, NR27, NR32, NR33, and Ara13

  Masato Nakanishi, Pedro Meirelles, Ryohei Suzuki, Naoki Takatani, Sayaka Mino, Wataru Suda, Kenshiro Oshima, Masahira Hattori, Moriya Ohkuma, Masashi Hosokawa, Kazuo Miyashita, Fabiano L. Thompson, Ako Niwa, Toko Sawabe, Tomoo Sawabe

  Genome Announcements 2 (6) 2169-8287 2014 [Refereed][Not invited]
   

  Here, we present the draft genome sequences of six carotenoid producers affiliated with Nonlabens spp. isolated from marine environments in both the northern and southern parts of Japan. The genomic information will help to elucidate the function and evolution of carotenoid synthetic gene clusters not only in the genus Nonlabens but also in the family Flavobacteriaceae.
• Draft genome sequences of marine flavobacterium Algibacter lectus strains SS8 and NR4

  Naoki Takatani, Masato Nakanishi, Pedro Meirelles, Sayaka Mino, Wataru Suda, Kenshiro Oshima, Masahira Hattori, Moriya Ohkuma, Masashi Hosokawa, Kazuo Miyashita, Fabiano L. Thompson, Ako Niwa, Toko Sawabe, Tomoo Sawabe

  Genome Announcements 2 (6) 2169-8287 2014 [Refereed][Not invited]
   

  Here, we present the draft genome sequences of a zeaxanthin-producing flavobacterium, Algibacter lectus strains SS8 and NR4, isolated from coastal sediment and rock surfaces in Hakodate, Japan, respectively. This genomic information represents the first Algibacter genome sequences, which will help us to elucidate the biology and evolution of Flavobacteriaceae bacteria.
• Draft genome sequence of marine flavobacterium Jejuia pallidilutea strain 11shimoA1 and pigmentation mutants

  Naoki Takatani, Masato Nakanishi, Pedro Meirelles, Sayaka Mino, Wataru Suda, Kenshiro Oshima, Masahira Hattori, Moriya Ohkuma, Masashi Hosokawa, Kazuo Miyashita, Fabiano L. Thompson, Ako Niwa, Toko Sawabe, Tomoo Sawabe

  Genome Announcements 2 (6) 2169-8287 2014 [Refereed][Not invited]
   

  Here, we present the draft genome sequence of a novel carotenoid 2'-isopentenylsaproxanthin producer, Jejuia pallidilutea strain 11shimoA1, isolated from the surface of seaweed in Japan, and the ethyl methanesulfonate-induced pigmentation mutants. This genomic information will help to not only elucidate the 2'-isopentenylsaproxanthin biosynthetic pathway but also understand the evolution of flavobacteria.
• Fucoxanthin: A Marine Carotenoid Exerting Anti-Cancer Effects by Affecting Multiple Mechanisms

  Sangeetha Ravi Kumar, Masashi Hosokawa, Kazuo Miyashita

  MARINE DRUGS 11 (12) 5130 - 5147 1660-3397 2013/12 [Refereed][Not invited]
   

  Fucoxanthin is a marine carotenoid exhibiting several health benefits. The anti-cancer effect of fucoxanthin and its deacetylated metabolite, fucoxanthinol, is well documented. In view of its potent anti-carcinogenic activity, the need to understand the underlying mechanisms has gained prominence. Towards achieving this goal, several researchers have carried out studies in various cell lines and in vivo and have deciphered that fucoxanthin exerts its anti-proliferative and cancer preventing influence via different molecules and pathways including the Bcl-2 proteins, MAPK, NFB, Caspases, GADD45, and several other molecules that are involved in either cell cycle arrest, apoptosis, or metastasis. Thus, in addition to decreasing the frequency of occurrence and growth of tumours, fucoxanthin has a cytotoxic effect on cancer cells. Some studies show that this effect is selective, i.e., fucoxanthin has the capability to target cancer cells only, leaving normal physiological cells unaffected/less affected. Hence, fucoxanthin and its metabolites show great promise as chemotherapeutic agents in cancer.
• Chemical and nutritional characteristics of brown seaweed lipids: A review

  Kazuo Miyashita, Nana Mikami, Masashi Hosokawa

  JOURNAL OF FUNCTIONAL FOODS 5 (4) 1507 - 1517 1756-4646 2013/10 [Refereed][Not invited]
   

  Several brown seaweeds show high total lipids (TL) contents, ranging from 10 to 20 wt.% per dry weight. The main lipid class is glycoglycerolipids (GL), which are rich in 18:4n-3, 20:5n-3 and 20:4n-6. Brown seaweed TL also contains fucoxanthin as a key functional compound. Despite the high levels of 18:4n-3, 20:5n-3 and 20:4n-6, brown seaweed lipids are stable to oxidation. Their high oxidative stability is partly related to the presence of these polyunsaturated fatty acids in their GL forms. Brown seaweed lipids show anti-obesity and anti-diabetic effects, which are mainly due to the up-regulatory effect of fucoxanthin on energy expenditure in abdominal white adipose tissue and glucose utilization in muscle. (C) 2013 Published by Elsevier Ltd.
• Oxidative Stability of Lipids Rich in EPA and DHA Extracted from Fermented Scallop Ovary

  Naohiro Hamaoka, Junki Shimajiri, Masayuki Abe, Masashi Hosokawa, Kazuo Miyashita

  JOURNAL OF FOOD SCIENCE 78 (9) C1348 - C1353 0022-1147 2013/09 [Refereed][Not invited]
   

  A novel seafood paste was developed by the fermentation of scallop ovary using rice malt (koji) and yeast culture. Chemical analysis of the product showed the formation of high level of free amino acids and organic acids during the fermentation. The product color and flavor resembled to Japanese traditional soybean miso. The contents of total lipids (TLs) extracted from the fermented products were ranged from 9.18% to 11.59% or 11.38% to 13.57%/dry sample weight. Although the TL was rich in oxidatively unstable polyunsaturated fatty acids (PUFAs) such as eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA), little decrease was found in these PUFAs during the fermentation, showing the high oxidative stability of the TL from the fermented scallop ovary. Moreover, the oxidative stability of the TL extracted from the fermented products increased with increasing the fermentation time. This would be mainly due to the formation of lipid soluble antioxidants such as tocopherols, which might be derived from yeast used for fermentation.
• Synergistic Antioxidant Activity of Milk Sphingomyeline and Its Sphingoid Base with alpha-Tocopherol on Fish Oil Triacylglycerol

  Junki Shimajiri, Makoto Shiota, Masashi Hosokawa, Kazuo Miyashita

  JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY 33 61 (33) 7969 - 7975 0021-8561 2013/08 [Refereed][Not invited]
   

  The effects of milk phospholipids (PLs), sphingolipids (SLs), and their sphingoid backbone on the oxidation of fish oil triacylglycerol (TAG) were examined with or without alpha-tocopherol. All compounds had little effect on the TAG oxidation in the absence of alpha-tocopherol. On the other hand, they could act synergistically with alpha-tocopherol. The highest synergistic activity was shown by sphingoid bases, followed by sphingomyelin (SPM) and other amine-containing PLs and SLs. This result showed that the synergistic activity increased with an increasing concentration of amine group of PLs, SLs, or sphingoid bases in the reaction mixture. The comparison of changes in alpha-tocopherol content in fish oil TAG and tricaprylin suggested that antioxidant compounds would be formed from the amine group and the lipid oxidation products in a mild oxidation condition controlled by alpha-tocopherol.
• Seasonal variations of total lipids, fatty acid composition, and fucoxanthin contents of Sargassum horneri (Turner) and Cystoseira hakodatensis (Yendo) from the northern seashore of Japan

  Masatoshi Nomura, Hiroyuki Kamogawa, Eko Susanto, Chikara Kawagoe, Hajime Yasui, Naotsune Saga, Masashi Hosokawa, Kazuo Miyashita

  JOURNAL OF APPLIED PHYCOLOGY 25 (4) 1159 - 1169 0921-8971 2013/08 [Refereed][Not invited]
   

  The present study describes the seasonal changes in lipid components of two brown algae, Sargassum horneri (Turner) and Cystoseira hakodatensis (Yendo), with specific reference to fucoxanthin (Fx) and n-3 (omega-3) polyunsaturated fatty acids (PUFAs). Young thalli of S. horneri were collected from different places, Nesaki and Matsushima, and cultivated in the same place. Total lipids (TL) and Fx contents of S. horneri increased from October or November and reached a maximum in January and then decreased thereafter. TL and Fx of S. horneri originated from Matsushima were always higher than those originated from Nesaki, suggesting that genetic variation might be a reason for the different TL and Fx contents found in both S. horneri seeds from different origin. Similar seasonal variation in the TL and Fx content has been more clearly observed in C. hakodatensis. Maximum TL content of S. horneri and C. hakodatensis reached to more than 14 and 15 % in winter season. These values were much higher than those found in previous studies showing TL content range from 1 to 5 %. Fatty acid composition of lipids from S. horneri and C. hakodatensis also changed seasonally. Total monounsaturated fatty acids showed the lower values from January to April, while there were no big variations in total saturated fatty acids and total n-6 PUFAs. On the other hand, total n-3 PUFAs became highest in winter. Overall, S. horneri and C. hakodatensis harvested from winter to spring will be used as good n-3 PUFA source with high levels of TL and Fx.
• Biosynthetic Pathway and Health Benefits of Fucoxanthin, an Algae-Specific Xanthophyll in Brown Seaweeds

  Koji Mikami, Masashi Hosokawa

  INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES 14 (7) 13763 - 13781 1422-0067 2013/07 [Refereed][Not invited]
   

  Fucoxanthin is the main carotenoid produced in brown algae as a component of the light-harvesting complex for photosynthesis and photoprotection. In contrast to the complete elucidation of the carotenoid biosynthetic pathways in red and green algae, the biosynthetic pathway of fucoxanthin in brown algae is not fully understood. Recently, two models for the fucoxanthin biosynthetic pathway have been proposed in unicellular diatoms; however, there is no such information for the pathway in brown seaweeds to date. Here, we propose a biosynthetic pathway for fucoxanthin in the brown seaweed, Ectocarpus siliculosus, derived from comparison of carotenogenic genes in its sequenced genome with those in the genomes of two diatoms, Thalassiosira pseudonana and Phaeodactylum tricornutum. Currently, fucoxanthin is receiving attention, due to its potential benefits for human health. Therefore, new knowledge regarding the medical and nutraceutical properties of fucoxanthin from brown seaweeds is also summarized here.
• Down-Regulation of Hepatic Stearoyl-CoA Desaturase-1 Expression by Fucoxanthin via Leptin Signaling in Diabetic/Obese KK-A (y) Mice

  Fumiaki Beppu, Masashi Hosokawa, Mi-Jin Yim, Taro Shinoda, Kazuo Miyashita

  LIPIDS 48 (5) 449 - 455 0024-4201 2013/05 [Refereed][Not invited]
   

  Stearoyl-coenzyme A desaturase-1 (SCD1) is a rate-limiting enzyme that catalyzes the biosynthesis of monounsaturated fatty acids from saturated fatty acids. Recently, SCD1 down-regulation has been implicated in the prevention of obesity, and the improvement of insulin and leptin sensitivity. In this study, we examined the effect of fucoxanthin, a marine carotenoid, on hepatic SCD1 in obese mouse models of hyperleptinemia KK-A (y) and leptin-deficiency ob/ob. In KK-A (y) mice, providing a diet containing 0.2 % fucoxanthin for 2 weeks markedly suppressed SCD1 mRNA and protein expressions in the liver. The fatty acid composition of liver lipids was also affected by an observed decrease in the ratio of oleic acid to stearic acid. Furthermore, serum leptin levels were significantly decreased in hyperleptinemia KK-A (y) mice after 2 weeks of fucoxanthin feeding. However, the suppressive effects of fucoxanthin on hepatic SCD1 and body weight gain were not observed in ob/ob mice. These results show that fucoxanthin down-regulates SCD1 expression and alters fatty acid composition of the liver via regulation of leptin signaling in hyperleptinemia KK-A (y) mice but not in leptin-deficient ob/ob mice.
• Substrate and Droplet Size: Important Factors for Understanding Aqueous Lipid Oxidation

  Kazuo Miyashita, Masashi Hosokawa

  Lipid Oxidation: Challenges in Food Systems 155 - 176 2013/04/02 [Refereed][Not invited]
   

  Lipid oxidation generally proceeds from the interface to the interior of the oil droplet in Oil-in-Water (O/W) emulsions therefore, the oxidation of lipids at the interface is an important factor to predict the oxidative stability of lipids in emulsion. In emulsions, the membranes surrounding the emulsion droplets consist of surface-active substances such as emulsifiers and/or proteins, together with lipids and water. They can sometimes provide a protective barrier for the penetration and diffusion of metals or radicals that initiate lipid oxidation. Therefore, the chemical and physical nature of the emulsifier and proteins is an important factor for the protection of PUFAs against oxidation in the aqueous phase. The different affinities of antioxidants and/or pro-oxidants to the interface are another important factor. Overall, the major factors affecting lipid oxidation at the interface are: affinity (partition) of antioxidants and/or pro-oxidants to the interface and chemical, physical, and stereo chemical susceptibility of interface lipids to oxidation. The behavior of antioxidants and pro-oxidants at the interface is related to their polarity and electrical charge. The oxidative stability of lipids at the interface is influenced by the size and concentration of the emulsion droplets, the thickness and the packing degree of interface molecules, and the extent of droplet-droplet interactions. Moreover, substrate lipid class and the degree of unsaturation have been reported to affect the oxidative characteristics at the interface. This chapter focuses on the effect of emulsifier, substrate lipids, and droplet size as a series of factors that affect lipid oxidation at the interface in O/W emulsion.
• Bovine lactoferrin reduces visceral fat and liver triglycerides in ICR mice

  Satoru Morishita, Tomoji Ono, Chikako Fujisaki, Yasuharu Ishihara, Michiaki Murakoshi, Hisanori Kato, Masashi Hosokawa, Kazuo Miyashita, Keikichi Sugiyama, Hoyoku Nishino

  JOURNAL OF OLEO SCIENCE 62 (2) 97 - 103 1345-8957 2013/02 [Refereed][Not invited]
   

  Lactoferrin (LF) is a multi-functional glycoprotein found in milk. In a previous clinical trial, we showed that enteric-coated bovine LF (bLF) tablets could reduce visceral fat accumulation. We also showed that bLF had anti-adipogenic activity in vitro. However, the mechanisms responsible for these phenomena remain unclear. In this study, we established an animal model of visceral fat reduction via oral bLF administration. We used gastric intubation to ensure that LF was absorbed in the small intestine. bLF administration for 4 weeks significantly reduced mesenteric fat tissue (P < 0.05) and hepatic triglyceride levels (P < 0.01). Furthermore, these two outcomes were positively correlated (R = 0.581, P < 0.05). Overall, these findings suggest that bLF affects mesenteric adipocytes and fatty acid metabolism in the liver.
• Chemopreventive Effects of Astaxanthin on Inflammatory Bowel Disease and Inflammation-Related Colon Carcinogenesis

  Masashi Hosokawa, Yumiko Yasui

  Carotenoids and Vitamin A in Translational Medicine 289 - 306 2013/01/01 

  Carotenoids are a family of natural pigments with at least 750 members. They possess a polyisoprenoid structure and are synthesized by plants, seaweeds, and microorganisms, but not by animals. Fruits and vegetables are the major sources of dietary carotenoids [1]. Yellow-orange vegetables and fruits provide a-and β-carotene, orange fruits provide β-cryptoxanthin, dark green leaves and vegetables.
• Fucoxanthin as potential nutraceutical for obesity therapy: A Review.

  Miyashita K, Maeda H, Nishikawa S, Tsukui T, Widjaja-Adhi MA-K, Hosokawa M

  Carotenoid Sci. 17 1 - 7 2013 [Refereed][Not invited]
  
• Effects of dietary fucoxanthin on cholesterol metabolism in diabetic/obese KK-A(y) mice

  Fumiaki Beppu, Masashi Hosokawa, Yoshimi Niwano, Kazuo Miyashita

  LIPIDS IN HEALTH AND DISEASE 11 112  1476-511X 2012/09 [Refereed][Not invited]
   

  Background: Fucoxanthin is a xanthophyll present in brown seaweeds and has several beneficial effects, including anti-obesity and anti-diabetic effects. However, we and another group previously observed that fucoxanthin increases serum cholesterol levels in rodents. Cholesterol is an important component of cell membranes and biosynthesis of bile acids. Serum cholesterol levels are also closely associated with atherosclerosis. Therefore, we sought to identify the mechanism underlying the increase in serum cholesterol levels by fucoxanthin. Methods: Diabetic/obese KK-A(y) mice were fed a diet containing 0.2% fucoxanthin for 4 weeks. The mice were sacrificed, and total blood samples were collected for the measurement of serum total cholesterol, HDL-cholesterol and non-HDL-cholesterol levels. Cholesterol content in tissues was also analyzed. Real-time PCR and Western blotting were performed to determine hepatic mRNA and protein expression of genes involved in cholesterol metabolism, respectively. Results: Dietary fucoxanthin significantly increased serum HDL and non-HDL cholesterol levels, and reduced hepatic cholesterol content. In liver, the expression of SREBP1, SREBP2 and their target genes involved in cholesterol biosynthesis significantly increased and tended to increase in the fucoxanthin-fed mice, respectively. In contrast, hepatic levels of LDLR and SR-B1 proteins which is important factors for LDL-cholesterol and HDL-cholesterol uptake in the liver from serum, decreased to 60% and 80% in the fucoxanthin-fed mice, respectively, compared with the control mice. Further, we found that dietary fucoxanthin significantly increased the mRNA expression of proprotein convertase subtilisin/kexin type 9 (PCSK9), which enhances intracellular degradation of LDLR in lysosomes. Conclusions: Fucoxanthin increased HDL-cholesterol and non-HDL-cholesterol levels in KK-A(y) mice by inducing SREBP expression and reduced cholesterol uptake in the liver via down-regulation of LDLR and SR-B1, resulted in increased serum cholesterol in the mice.
• Oxidative Stability of Glyceroglycolipids Containing Polyunsaturated Fatty Acids

  Tomoyo Yamaguchi, Rumiko Sugimura, Junki Shimajiri, Masatake Suda, Masayuki Abe, Masashi Hosokawa, Kazuo Miyashita

  JOURNAL OF OLEO SCIENCE 61 (9) 505 - 513 1345-8957 2012/09 [Refereed][Not invited]
   

  The oxidative stability of glyceroglycolipids (GLs) from spinach or brown edible seaweed (Laminaria japonica) was compared with those of phosphatidylcholines (PCs) from salmon roe and triacyglycerols (TAGs) from soybean oil or sardine oil. All the lipids were subjected to autoxidation after removing oxidants and/or antioxidants such as chlorophylls, tocopherols, and carotenoids. The oxidative stability of the lipids decreased with increasing number of bisallylic positions in the molecule. Due to the higher mean number of bisallylic positions, salmon roe PC and sardine oil TAG were oxidized more rapidly than soybean oil TAG. Spinach GL and brown edible seaweed GL showed the same oxidative stability as that found in soybean oil TAG, although the mean number of bisallylic positions of both GLs was much higher than that of soybean oil TAG and approached the number found in sardine oil TAG and salmon roe PC. The present study indicates the important effect of galactosyl and sulfoquinovosyl moieties on the oxidative stability of GL.
• Effect of Caffeine and Capsaicin on the Blood Glucose Levels of Obese/Diabetic KK-A(gamma) Mice

  Tomoyuki Okumura, Takayuki Tsukui, Masashi Hosokawa, Kazuo Miyashita

  JOURNAL OF OLEO SCIENCE 61 (9) 515 - 523 1345-8957 2012/09 [Refereed][Not invited]
   

  The present study is aimed to measure the effect of caffeine and capsaicin on the blood glucose level of obese/diabetic model mice. The blood glucose level of KK-A(gamma) obese/diabetic mice decreased significantly after dietary supplementations with less than 0.031% caffeine and less than 0.0042% capsaicin, while both food ingredients and the combination had little effect on body weight gain and abdominal white adipose tissue (WAT) weight at this dose. The blood glucose level of the control mice was 234.4 mg/dL at 10 days after feeding and it increased to 500.6 mg/dL after 28 days, while levels after intake of caffeine, capsaicin, and both together increased from 180, 131.6, and 140 mg/dL to 183.7, 255.1, and 151.6 mg/dL, respectively. The anti-diabetic activity of caffeine and capsaicin was confirmed by a decrease in total drinking water and in the blood insulin level.
• Stability of Fucoxanthin in Dried Undaria Pinnatifida (Wakame) and Baked Products (Scones) Containing Wakame Powder

  Rumiko Sugimura, Masatake Suda, Ayumi Sho, Tomoya Takahashi, Tokutake Sashima, Masayuki Abe, Masashi Hosokawa, Kazuo Miyashita

  FOOD SCIENCE AND TECHNOLOGY RESEARCH 18 (5) 687 - 693 1344-6606 2012/09 [Refereed][Not invited]
   

  Stability of fucoxanthin was assessed in dried Undaria pinnatifida (wakame). After 210 days storage of dried wakame in the dark at 50 degrees C, the stability of fucoxanthin was higher than that of purified fucoxanthin incubated at both 20 degrees C and 40 degrees C. Furthermore, we analyzed the stability of fucoxanthin in scones containing 0.5 - 2 wt% commercial dried wakame powder. More than 85% of the fucoxanthin remained unchanged in the scones after heating the dough at 190 degrees C for 30 min. Sensory analysis indicated that although the addition of wakame powder darkened the appearance and strengthened both the taste and smell of the scones, they remained acceptable. Furthermore, overall sensory scores showed that scones containing 0.5 wt% and 2 wt% wakame powder were more favorable than control scones.
• Effects of dietary fucoxanthin on cholesterol metabolism in diabetic/obese KK-A(y) mice

  Fumiaki Beppu, Masashi Hosokawa, Yoshimi Niwano, Kazuo Miyashita

  LIPIDS IN HEALTH AND DISEASE 11 112  1476-511X 2012/09 [Refereed][Not invited]
   

  Background: Fucoxanthin is a xanthophyll present in brown seaweeds and has several beneficial effects, including anti-obesity and anti-diabetic effects. However, we and another group previously observed that fucoxanthin increases serum cholesterol levels in rodents. Cholesterol is an important component of cell membranes and biosynthesis of bile acids. Serum cholesterol levels are also closely associated with atherosclerosis. Therefore, we sought to identify the mechanism underlying the increase in serum cholesterol levels by fucoxanthin.Methods: Diabetic/obese KK-A(y) mice were fed a diet containing 0.2% fucoxanthin for 4 weeks. The mice were sacrificed, and total blood samples were collected for the measurement of serum total cholesterol, HDL-cholesterol and non-HDL-cholesterol levels. Cholesterol content in tissues was also analyzed. Real-time PCR and Western blotting were performed to determine hepatic mRNA and protein expression of genes involved in cholesterol metabolism, respectively.Results: Dietary fucoxanthin significantly increased serum HDL and non-HDL cholesterol levels, and reduced hepatic cholesterol content. In liver, the expression of SREBP1, SREBP2 and their target genes involved in cholesterol biosynthesis significantly increased and tended to increase in the fucoxanthin-fed mice, respectively. In contrast, hepatic levels of LDLR and SR-B1 proteins which is important factors for LDL-cholesterol and HDL-cholesterol uptake in the liver from serum, decreased to 60% and 80% in the fucoxanthin-fed mice, respectively, compared with the control mice. Further, we found that dietary fucoxanthin significantly increased the mRNA expression of proprotein convertase subtilisin/kexin type 9 (PCSK9), which enhances intracellular degradation of LDLR in lysosomes.Conclusions: Fucoxanthin increased HDL-cholesterol and non-HDL-cholesterol levels in KK-A(y) mice by inducing SREBP expression and reduced cholesterol uptake in the liver via down-regulation of LDLR and SR-B1, resulted in increased serum cholesterol in the mice.
• Dietary Combination of Fish Oil and Taurine Decreases Fat Accumulation and Ameliorates Blood Glucose Levels in Type 2 Diabetic/Obese KK-A(y) Mice

  Nana Mikami, Masashi Hosokawa, Kazuo Miyashita

  JOURNAL OF FOOD SCIENCE 77 (6) H114 - H120 0022-1147 2012/06 [Refereed][Not invited]
   

  n-3 Polyunsaturated fatty acids (PUFAs) such as eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA), and taurine are functional compounds abundantly present in seafoods. In this study, we examined the combined effects of EPA- and DHA-rich fish oil and taurine on white adipose tissue (WAT) weight and blood glucose levels in diabetic/obese KK-A(y) mice. After a 4-wk administration of experimental diets (soybean oil or fish oil, supplemented with 0%, 2%, or 4% taurine), the increase in WAT weight of the mice fed the "fish oil + 4% taurine" diet was significantly suppressed compared to the "soybean oil + 4% taurine" and "fish oil only" diets. Serum triglycerides, free fatty acids, and total cholesterol levels decreased by fish oil administration. In addition, fish oil and taurine increased the activity of acyl-CoA oxidase, which is the rate-limiting enzyme of peroxisomal beta-oxidation, increased in the liver of KK-A(y) mice. The activity of fatty acid synthase decreased by fish oil diets. Furthermore, blood glucose and insulin levels were significantly lower in the mice fed fish oil than in the soybean oil-fed mice. In fish oil + 4% taurine group, hyperglycemia and hyperinsulinemia were effectively improved in KK-A(y) mice compared to the fish oil only groups. In particular, the combination of fish oil and taurine enhanced the glucose transporter 4 (GLUT4) distribution in the plasma membrane of muscle tissue. These results suggest that EPA- and DHA-rich fish oil, especially in combination with taurine, exhibits preventive effects on WAT weight gain and hyperglycemia in diabetic/obese KK-A(y) mice.
• Fucoxanthin promotes translocation and induction of glucose transporter 4 in skeletal muscles of diabetic/obese KK-A(y) mice

  Sho Nishikawa, Masashi Hosokawa, Kazuo Miyashita

  PHYTOMEDICINE 19 (5) 389 - 394 0944-7113 2012/03 [Refereed][Not invited]
   

  Fucoxanthin (Fx) isolated from Undaria pinnatifida suppresses the development of hyperglycemia and hyperinsulinemia of diabetic/obese KK-A(y) mice after 2 weeks of feeding 0.2% Fx-containing diet. In the soleus muscle of KK-A(y) mice that were fed Fx, glucose transporter 4 (GLUT4) translocation to plasma membranes from cytosol was promoted. On the other hand, Fx increased GLUT4 expression levels in the extensor digitorum longus (EDL) muscle, although GLUT4 translocation tended to increase. The expression levels of insulin receptor (IR) mRNA and phosphorylation of Akt, which are in upstream of the insulin signaling pathway regulating GLUT4 translocation, were also enhanced in the soleus and EDL muscles of the mice fed Fx. Furthermore, Fx induced peroxisome proliferator activated receptor gamma coactivator-1 alpha (PGC-1 alpha), which has been reported to increase GLUT4 expression, in both soleus and EDL muscles. These results suggest that in diabetic/obese KK-A(y) mice, Fx improves hyperglycemia by activating the insulin signaling pathway, including GLUT4 translocation, and inducing GLUT4 expression in the soleus and EDL muscles, respectively, of diabetic/obese KK-A(y) mice. (C) 2011 Elsevier GmbH. All rights reserved.
• Therapeutic Effect of Fucoxanthin on Metabolic Syndrome and Type 2 Diabetes

  Kazuo Miyashita, Show Nishikawa, Masashi Hosokawa

  Nutritional and Therapeutic Interventions for Diabetes and Metabolic Syndrome 367 - 379 2012 [Not refereed][Not invited]
  
• Analyses on the functions and developments for the utilization technology of health functional components derived from fisheries products -Effective utilization and development for the next stage-

  Hideki Ushio, Naohiro Gotoh, Reiko Nagasaka, Masashi Hosokawa

  Nippon Suisan Gakkaishi (Japanese Edition) 78 (5) 1053 - 1056 0021-5392 2012 [Not refereed][Not invited]
  
• Carotenoid Profile of Edible Japanese Seaweeds: An Improved HPLC Method for Separation of Major Carotenoids

  Masaru Terasaki, Bhaskar Narayan, Hiroyuki Kamogawa, Masatoshi Nomura, Nimish Mol Stephen, Chikara Kawagoe, Masashi Hosokawa, Kazuo Miyashita

  JOURNAL OF AQUATIC FOOD PRODUCT TECHNOLOGY 21 (5) 468 - 479 1049-8850 2012 [Refereed][Not invited]
   

  An improved multi-step gradient reverse phase high performance liquid chromatography (HPLC) method to simultaneously separate major carotenoids from natural as well as food samples was developed. Quantitative profiling of carotenoid compounds was carried out on three edible brown seaweeds (Sargassum horneri, Cystoseira hakodatensis, and Undaria pinnatifida) and three red seaweeds (Gracilaria vermiculophylla, Grateloupia asiatica, and Grateloupia livida). Fucoxanthin (Fx) was detected in all the brown seaweeds with quantities (mg g-1 dry weight [dwt]) ranging from 1.3 +/- 0.3 in C. hakodatensis to 2.4 +/- 0.1 in S. horneri. U. pinnatifida, commonly known as wakame, had a Fx content of 2.3 +/- 0.1 mg g-1 dwt. In the case of red seaweeds, zeaxanthin (Zx) was the major carotenoid, and G. vermiculophylla had the highest Zx content (80.2 mu g g-1 dwt) among the red seaweeds apart from small amounts of Fx (9.1 mu g g-1 dwt). Similarly, the other two species of red seaweeds, G. asiatica and G. livida, contained (mu g g-1 dwt) lutein (Lut), Fx, and Zx as the major carotenoids (G. asiatica: Lut 10.4, Fx 1.5, Zx 1.1; G. livida: Lut 9.3, Fx 3.5, Zx 1.0). The results suggest the usefulness of edible varieties (barring wakame) of seaweeds as dietary sources of carotenoids.
• In vivo antioxidant activity of fucoxanthin on obese/diabetes KK-Ay mice.

  S. Iwasaki, M. A-K. Widjaja-Adhi, A. Koide, T. Kaga, S. Nakano, F. Beppu, M. Hosokawa, K. Miyashita

  Food Nutr. Sci. 3 1491 - 1499 2012 [Refereed][Not invited]
  
• Brown Seaweed Lipids as Potential Source of Omega-3 PUFA in Biological Systems

  Kazuo Miyashita, Bhaskar Narayan, Takayuki Tsukui, Hiroyuki Kamogawa, Masayuki Abe, Masashi Hosokawa

  Handbook of Marine Macroalgae: Biotechnology and Applied Phycology 329 - 339 2011/11/21 [Not refereed][Not invited]
  
• Algal Carotenoids as Potent Antioxidants

  Kazuo Miyashita, M. Airanthi K. Widjaja-Adhi, Masayuki Abe, Masashi Hosokawa

  Handbook of Marine Macroalgae: Biotechnology and Applied Phycology 403 - 414 2011/11/21 [Not refereed][Not invited]
  
• Cancer Chemopreventive Ability of Conjugated Linolenic Acids

  Takuji Tanaka, Masashi Hosokawa, Yumiko Yasui, Rikako Ishigamori, Kazuo Miyashita

  INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES 12 (11) 7495 - 7509 1422-0067 2011/11 [Refereed][Not invited]
   

  Conjugated fatty acids (CFA) have received increased interest because of their beneficial effects on human health, including preventing cancer development. Conjugated linoleic acids (CLA) are such CFA, and have been reviewed extensively for their multiple biological activities. In contrast to other types of CFAs including CLA that are found at low concentrations (less than 1%) in natural products, conjugated linolenic acids (CLN) are the only CFAs that occur in higher quantities in natural products. Some plant seeds contain a considerably high concentration of CLN (30 to 70 wt% lipid). Our research group has screened CLN from different plant seed oils to determine their cancer chemopreventive ability. This review describes the physiological functions of CLN isomers that occur in certain plant seeds. CLN are able to induce apoptosis through decrease of Bcl-2 protein in certain human cancer cell lines, increase expression of peroxisome proliferator-activated receptor (PPAR)-gamma, and up-regulate gene expression of p53. Findings in our preclinical animal studies have indicated that feeding with CLN resulted in inhibition of colorectal tumorigenesis through modulation of apoptosis and expression of PPAR gamma and p53. In this review, we summarize chemopreventive efficacy of CLN against cancer development, especially colorectal cancer.
• The effect of milk polar lipids separated from butter serum on the lipid levels in the liver and the plasma of obese-model mouse (KK-A(y))

  Shiomi Watanabe, Tomoki Takahashi, Leo Tanaka, Yuko Haruta, Makoto Shiota, Masashi Hosokawa, Kazuo Miyashita

  JOURNAL OF FUNCTIONAL FOODS 3 (4) 313 - 320 1756-4646 2011/10 [Refereed][Not invited]
   

  The effect of lipid concentrated-butter serum (LC-BS) on the plasma and hepatic cholesterol and triacylglycerol levels of obese-model mouse (KK-A(Y)) was evaluated. The same analysis was done on the main lipid fractions from LC-BS, namely, ceramide rich fraction (Ceramide-fr), phosphatidylethanolamine (PE) rich fraction (PE-fr), and sphingomyelin (SPM) rich fraction (SPM-fr). Plasma cholesterol of mice fed LC-BS and Ceramide-fr was significantly (P < 0.05) lower than that of the control. Hepatic total cholesterol and triacylglycerol levels was also reduced significantly by Ceramide-fr feeding (P < 0.05). Although there was no significant difference in the fatty acid composition of all dietary lipids, significant decrease (P < 0.05) in content of 18:1n-9 was found in the liver lipids of the mice fed Ceramide-fr containing diet. The level of 16:1n-7 and 18:1n-7 was also reduced by Ceramide-fr feeding. This result suggests the down-regulation of stearoyl-CoA desaturase-1 (SCD1) by milk ceramides, which would be related to the decrease in triacylglycerol and cholesterol levels in the liver and in the plasma. (C) 2011 Elsevier Ltd. All rights reserved.
• Nitrocapsanthin and Nitrofucoxanthin, Respective Products of Capsanthin and Fucoxanthin Reaction with Peroxynitrite

  Makoto Tsuboi, Hideo Etoh, Kyuki Kato, Hiroki Nakatugawa, Hideki Kato, Yasunori Maejima, Gaku Matsumoto, Hironobu Mori, Masashi Hosokawa, Kazuo Miyashita, Harukuni Tokuda, Nobukuni Suzuki, Takashi Maoka

  JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY 59 (19) 10572 - 10578 0021-8561 2011/10 [Refereed][Not invited]
   

  The in vitro reactivity of capsanthin (1) and fucoxanthin (2) with peroxynitrite was investigated, and the reaction products produced by scavenging with peroxynitrite were analyzed. (14'Z)-Nitrocapsanthin (3) and 12-nitrocapsanthin (4) were isolated from the products of the reaction of capsanthin with peroxynitrite. Similarly, (14Z)-15-nitrofucoxanthin (5), (11Z)-11-nitrofucoxanthin (6), and (14Z,9'Z)-15-nitrofucoxanthin (7) were obtained from the reaction of peroxynitrite reaction with fucoxanthin. Capsanthin and fucoxanthin inhibited the nitration of tyrosine by peroxynitrite. Furthermore, nitrocapsanthins (3 and 4) and nitrofucoxanthins (5 and 6) exhibited an inhibitory effect on Epstein Barr virus early antigen activation in Raji cells and an antiproliferative effect on human pancreatic carcinoma. Moreover, nitrocapsanthins (3 and 4) inhibited carcinogensis of mouse skin tumors initiated by 7,12-dimethylbenz[a]anthracene (DMBN).
• Dietary astaxanthin inhibits colitis and colitis-associated colon carcinogenesis in mice via modulation of the inflammatory cytokines

  Yumiko Yasui, Masashi Hosokawa, Nana Mikami, Kazuo Miyashita, Takuji Tanaka

  CHEMICO-BIOLOGICAL INTERACTIONS 193 (1) 79 - 87 0009-2797 2011/08 [Refereed][Not invited]
   

  Astaxanthin (AX) is one of the marine carotenoid pigments, which possess powerful biological antioxidant, anti-inflammatory and anti-cancer properties. The purpose of this study is to investigate possible inhibitory effect of AX against inflammation-related mouse colon carcinogenesis and dextran sulfate sodium (DSS)-induced colitis in male ICR mice. We conducted two different experiments. In the first experiment, we evaluated the effects of AX at three dose levels, 50, 100 and 200 ppm in diet, on colitis-associated colon carcinogenesis induced by azoxymethane (AOM)/DSS in mice. In the second, the effects of the AX (10(1 and 200 ppm) in diet on DSS-induced colitis were determined. We found that dietary AX significantly inhibited the occurrence of colonic mucosal ulcers, dysplastic crypts, and colonic adenocarcinoma at week 20. AX-feeding suppressed expression of inflammatory cytokines, including nuclear factor (NF)-kappa B, tumor necrosis factor (TINIF)-alpha and interleukin (IL)-1 beta, inhibited proliferation, and induced apoptosis in the colonic adenocarcinomas. Feeding with 200 ppm AX, but not 100 ppm, significantly inhibited the development of DSS-induced colitis. AX feeding (200 ppm in diet) also lowered the protein expression of NF-kappa B, and the mRNA expression of inflammatory cytokines, including IL-1 beta, IL-6, and cyclooxygenase (COX)-2. Our results suggest that the dietary AX suppresses the colitis and colitis-related colon carcinogenesis in mice, partly through inhibition of the expression of inflammatory cytokine and proliferation. Our findings suggest that AX is one of the candidates for prevention of colitis and inflammation-associated colon carcinogenesis in humans. (C) 2011 Elsevier Ireland Ltd. All rights reserved.
• The allenic carotenoid fucoxanthin, a novel marine nutraceutical from brown seaweeds

  Kazuo Miyashita, Sho Nishikawa, Fumiaki Beppu, Takayuki Tsukui, Masayuki Abe, Andmasashi Hosokawa

  JOURNAL OF THE SCIENCE OF FOOD AND AGRICULTURE 91 (7) 1166 - 1174 0022-5142 2011/05 [Refereed][Not invited]
   

  Obesity and type 2 diabetes are pathologies with rapidly growing prevalence throughout the world. A few molecular targets offer the mosthope for anti-obesity and anti-diabetic therapeutics. One of the keys to success will be the induction of uncoupling protein 1 (UCP1) in abdominal white adipose tissue (WAT) and the regulation of cytokine secretions from both abdominal adipose cells and macrophage cells infiltrated into adipose tissue. Anti-obesity and anti-diabetic effects of fucoxanthin, a characteristic carotenoid found in brown seaweeds, have been reported. Nutrigenomic studies reveal that fucoxanthin induces UCP1 in abdominal WAT mitochondria, leading to the oxidation of fatty acids and heat production in WAT. Fucoxanthin improves insulin resistance and decreases blood glucose levels through the regulation of cytokine secretions from WAT. The key structure of carotenoids for the expression of anti-obesity effect is suggested to be the carotenoid end of the polyene chromophore, which contains an allenic bond and two hydroxyl groups. (C) 2011 Society of Chemical Industry
• Effect of Brown Seaweed Lipids on Fatty Acid Composition and Lipid Hydroperoxide Levels of Mouse Liver

  M. K. Widjaja-Adhi Airanthi, Naoya Sasaki, Sayaka Iwasaki, Nobuko Baba, Masayuki Abe, Masashi Hosokawa, Kazuo Miyashita

  JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY 59 (8) 4156 - 4163 0021-8561 2011/04 [Refereed][Not invited]
   

  Brown seaweed lipids from Undaria pinnahfida (Wakame), Sargassum horneri (Akamoku), and Cystoseira hakodatensis (Uganomoku) contained several bioactive compounds, namely, fucoxanthin, polyphenols, and omega-3 polyunsaturated fatty acids (PUPA). Fucoxanthin and polyphenol contents of Akamoku and Uganomoku lipids were higher than those of Wakame lipids, while Wakame lipids showed higher total omega-3 PUFA content than Akamoku and Uganomolcu lipids. The levels of docosahexaenoic acid (DHA) and arachidonic acid (AA) in liver lipids of KK-A(y) mouse significantly increased by Akamoku and Uganomoku lipid feeding as compared with the control, but not by Wakame lipid feeding. Fucoxanthin has been reported to accelerate the bioconversion of omega-3 PUFA and omega-6 PUPA to DHA and AA, respectively. The higher hepatic DHA and AA level of mice fed Akamoku and Uganomoku lipids would be attributed to the higher content of fucoxanthin of Akamoku and Uganomoku lipids. The lipid hydroperoxide levels of the liver of mice fed brown seaweed lipids were significantly lower than those of control mice, even though total PUFA content was higher in the liver of mice fed brown seaweed lipids. This would be, at least in part, due to the antioxidant activity of fucoxanthin metabolites in the liver.
• 褐藻由来フコキサンチンによるstearoyl-CoA desaturase 1(SCD1)発現抑制効果

  篠田 太郎, 別府 史章, 津久井 隆行, 細川 雅史, 宮下 和夫

  日本栄養・食糧学会大会講演要旨集 (公社)日本栄養・食糧学会 65回 163 - 163 2011/04
• Suppressive Effects of Amarouciaxanthin A on 3T3-L1 Adipocyte Differentiation through Down-regulation of PPAR gamma and C/EBP alpha mRNA Expression

  Mi-Jin Yim, Masashi Hosokawa, Yoshiyuki Mizushina, Hiromi Yoshida, Yasunori Saito, Kazuo Miyashita

  JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY 59 (5) 1646 - 1652 0021-8561 2011/03 [Refereed][Not invited]
   

  Fucoxanthin is converted to fucoxanthinol and amarouciaxanthin A in mice. It was previously reported that fucoxanthinol attenuated the adipogenesis of 3T3-L1 cells. However, the effects of amarouciaxanthin A on adipocyte differentiation have not been clarified. This study examined the effects of amarouciaxanthin A on 3T3-L1 adipogenesis by comparing the effects of fucoxanthinol, isofucoxanthinol, and amarouciaxanthin B. Amarouciaxanthin A significantly decreased glycerol-3-phosphate dehydrogenase (GPDH) activity, which was measured as an indicator of adipocyte differentiation. The suppressive effect of amarouciaxanthin A was stronger than that of fucoxanthinol, amarouciaxanthin B, and isofucoxanthinol. The mRNA expressions of adipocyte fatty acid-binding protein (aP2), lipoprotein lipase (LPL), and glucose-transporter 4 (Glut4) in 3T3-L1 cells were markedly down-regulated by amarouciaxanthin A compared to fucoxanthinol. Furthermore, the expression levels of peroxisome proliferator-activated receptor gamma (PPAR gamma) and CCAAT-enhancer-binding protein alpha (C/EBP alpha) which are the key adipogenic transcriptional factors, were also de creased by amarouciaxanthin A during adipocyte differentiation. These results show that amarouciaxanthin A, which is a dominant metabolite of fucoxanthin in white adipose tissue, suppressed 3T3-L1 adipocyte differentiation.
• Synthesis of novel phospholipids that bind phenylalkanols and hydroquinone via phospholipase D-catalyzed transphosphatidylation

  Yukihiro Yamamoto, Hideyuki Kurihara, Kazuo Miyashita, Masashi Hosokawa

  NEW BIOTECHNOLOGY 28 (1) 1 - 6 1871-6784 2011/01 [Refereed][Not invited]
   

  Phenylalkanols such as tyrosol and hydroxytyrosol (h-tyrosol), which possess antioxidant and anticancer properties, were phosphatidylated by phospholipase D (PLD)-catalyzed transphosphatidylation. After a 24-hour reaction of phosphatidylcholine (PC) and tyrosol with PLD, a new product was detected by TLC and identified to phosphatidyl-tyrosol by high-resolution MS and NMR analyses. The optimum reaction conditions were as follows: soyPC 50 mu mol, tyrosol 500 mu mol, ethyl acetate 1.6 ml, PLD 1.6 U, 0.2 M sodium acetate buffer (pH 5.6) 0.8 ml, 37 degrees C for 24 hours. Under the optimum reaction conditions, the yields of phosphatidyl-tyrosol, hydroquinone (HQ), 2-(4-aminophenyl) ethanol (4APE), h-tyrosol and 2-phenylethanol (PEA) were 87 +/- 3.7, 13 +/- 1.3, 90 +/- 2.3, 64 +/- 5.5 and 85 +/- 1.0 mol%, respectively. Furthermore, from the results of transphosphatidylation of soyPC with several phenylethanols and phenylpropanols, we established the following details about the reaction specificity of transphosphatidylation by PLD from Streptomyces sp.: (1) para-amino and para-hydroxyl groups in the benzene ring of PEA derivatives do not affect the transphosphatidylation by PLD, whereas meta-hydroxyl group slightly inhibits the transphosphatidylation. (2) Meta- and ortho-methyl groups in the benzene ring of PEA derivatives also slightly inhibit the transphosphatidylation. (3) Secondary and tertiary alcohols and hydroquinone are difficult to transphosphatidylate by PLD.
• Comparative Antioxidant Activity of Edible Japanese Brown Seaweeds

  M. K. Widjaja-Adhi Airanthi, Masashi Hosokawa, Kazuo Miyashita

  JOURNAL OF FOOD SCIENCE 76 (1) C104 - C111 0022-1147 2011/01 [Refereed][Not invited]
   

  Japanese edible brown seaweeds, Eisenia bicyclis (Arame), Kjellmaniella crassifolia (Gagome), Alaria crassifolia (Chigaiso), Sargassum horneri (Akamoku), and Cystoseira hakodatensis (Uganomoku) were assayed for total phenolic content (TPC), fucoxanthin content, radical scavenging activities (DPPH, peroxyl radical, ABTS, and nitric oxide), and antioxidant activity in a liposome system. Among the solvents used for extraction, methanol was the most effective to extract total phenolics (TPC) from brown seaweeds. Among 5 kinds of brown seaweeds analyzed, methanol extract from C. hakodatensis was the best source for antioxidants. The high antioxidant activity of the extract was based not only on the high content of phenolics, but on the presence of fucoxanthin. No significant correlation (P > 0.05) was observed between TPC per gram extract with DPPH radical scavenging activity of the methanol extracts. These observed discrepancy would be due to structural variations in the phenolic compounds, and different levels of fucoxanthin in the extracts. The present study also demonstrated the synergy in the antioxidant activity of the combination of brown seaweed phenolics and fucoxanthin.
• Antiobesity Effects of Undaria Lipid Capsules Prepared with Scallop Phospholipids

  Tomoko Okada, Yasuyuki Mizuno, Shinichi Sibayama, Masashi Hosokawa, Kazuo Miyashita

  JOURNAL OF FOOD SCIENCE 76 (1) H2 - H6 0022-1147 2011/01 [Refereed][Not invited]
   

  Based on previous research findings, a capsule was developed containing n-3 polyunsaturated fatty acid rich scallop phospholipids (PLs) with an incorporation of brown seaweed (Undaria pinnatifida) lipids (ULs) containing fucoxanthin. The antiobesity effects of the capsules were evaluated with an animal model using 3-wk-old male KK-A(y) mice. Each group received different combinations of lipid (UL, PL, UL + PL, or UL + PL capsule) either incorporated into the diet or into drinking water. Animals were sacrificed after a 4-wk experimental feeding period, and adipose tissues and organs were dissected and weighed. Blood samples were obtained to determine plasma lipid profiles. Uncoupling protein 1 (UCP1) mRNA expression levels were determined by real-time polymerase chain reaction analysis, and UCP1 expression was determined by western blotting analysis. Treatment with either UL alone or UL + PL (capsule) through drinking water resulted in a significant reduction in body weight, compared to the control group. The total white adipose tissue weight of mice fed the UL + PL capsule in drinking water was significantly reduced. Both UCP1 and UCP1 mRNA expression in epididymal fat from mice fed the capsule were significantly higher than in the control group. These results suggest that incorporation of UL into scallop-derived PL by means of capsulation may lead to an additive increase in the antiobesity properties of these bioactive lipids.
• Fucoxanthin regulates adipocytokine mRNA expression in white adipose tissue of diabetic/obese KK-A(y) mice

  Masashi Hosokawa, Tatsuya Miyashita, Sho Nishikawa, Shingo Emi, TakaYuki Tsukui, Fumiaki Beppu, Tomoko Okada, Kazuo Miyashita

  ARCHIVES OF BIOCHEMISTRY AND BIOPHYSICS 504 (1) 17 - 25 0003-9861 2010/12 [Refereed][Not invited]
   

  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-a (TNF-alpha), 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-a 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-alpha-induced MCP-1 and IL-6 mRNA overexpression and protein secretion into the culture medium. In addition, fucoxanthinol decreased TNF-a, 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. (C) 2010 Elsevier Inc. All rights reserved.
• Anti-obesity and anti-diabetic effects of allenic carotenoid, fucoxanthin

  Kazuo Miyashita, Hayato Maeda, Tomoko Okada, Masayuki Abe, Masashi Hosokawa

  AGRO FOOD INDUSTRY HI-TECH 21 (6) 24 - 27 1722-6996 2010/11 [Not refereed][Not invited]
   

  Fucoxanthin, a characteristic carotenoid found in brown seaweeds, shows anti-obesity and anti-diabetic effects on the basis of specific molecular mechanism. Fucoxanthin induces uncoupling protein 1 expression in white adipose tissue (WAT) mitochondria leading to oxidation of fatty acids and heat production in WAT. Fucoxanthin improves insulin resistance and decreases blood glucose level through the down-regulation of adipokines such as TNF alpha, MCP-1, IL-6, and PAI-1 in WAT and up-regulation of glucose transporter 4 (GLUT4) in skeletal muscle. The anti-obesity effect of fucoxanthin is likely linked to its structural characteristic - an allene bond and an additional hydroxyl substituent on the side group of the fucoxanthin metabolites, fucoxanthinol and amarouciaxanthin A.
• Value Addition to Seafood Processing Discards

  Sachindra M. Nakkarike, Bhaskar Narayan, Masashi Hosokawa, Kazuo Miyashita

  Handbook of Seafood Quality, Safety and Health Applications 390 - 401 2010/09/02 [Not refereed][Not invited]
  
• Scavenging of Active Nitrogene Species by Carotenoid : Reaction Products of Carotenoids with Peroxynitrite

  Etoh Hideo, Maoka Takashi, Kato Kyuki, Nakatsugawa Hiroki, Tsuboi Makoto, Terada Yukimasa, Matsumoto Gaku, Mori Hironobu, Hosokawa Masashi, Miyashita Kazuo

  Symposium on the Chemistry of Natural Products, symposium papers 天然有機化合物討論会 (52) 499 - 504 2010/09/01 

  Peroxynitrite is a powerful oxidant produced by macrophages and neutrophils. We have investigaed the reaction of peroxynitrite with some carotenoids such as β-carotene, astaxanthin and lutein in vitro. In the present study, reactions of peroxynitrite with capsanthin and fucoxanthin were studied. 14'-s-cia-15'-nitrocapsanthin (1), 12-nitro-capsanthin (2), 14-s-cis-15-nitrofucoxanthin (3), and 11-cis-11-nitro-fucoxanthin (4) were obtained by these reactions. These results indicated that carotenoids could take up peroxynitirite through the formation of nitrocarotenoids and/or oxazine carotenoids (in the case of lutein). These reactions would probably be occurred in vivo and suggest that carotenoids might contribute to the prevention of peroxynitrite induced disease.
• Purified Canola Lutein Selectively Inhibits Specific Isoforms of Mammalian DNA Polymerases and Reduces Inflammatory Response

  Sho Horie, Chiaki Okuda, Takatoshi Yamashita, Kenichi Watanabe, Kouji Kuramochi, Masashi Hosokawa, Toshifumi Takeuchi, Makiko Kakuda, Kazuo Miyashita, Fumio Sugawara, Hiromi Yoshida, Yoshiyuki Mizushina

  LIPIDS 45 (8) 713 - 721 0024-4201 2010/08 [Refereed][Not invited]
   

  In the screening of DNA polymerase (pol) inhibitor, we isolated lutein, a carotenoid, from the crude (unrefined) pressed oil of canola (low erucic acid rapeseed, Brassica napus L.). Commercially prepared carotenoids such as lutein (1), zeaxanthin (2), beta-cryptoxanthin (3), astaxanthin (4), canthaxanthin (5), beta-carotene (6), lycopene (7), capsanthin (8), fucoxanthin (9) and fucoxanthinol (10), were investigated for the inhibitory activities of pols. Compounds 1, 2 and 8 exhibited strong inhibition of the activities of mammalian pols beta and lambda, which are DNA repair- and/or recombination-related pols. On the other hand, all carotenoids tested had no influence on the activity of a mammalian pol alpha, which is a DNA replicative pol. Lutein (1) was the strongest pol inhibitor of mammalian pols beta and lambda in the prepared ten carotenoids tested, but did not influence of the activities of mammalian pols alpha, gamma, delta and epsilon. The tendency for pols beta and lambda inhibition by these carotenoids showed a positive correlation with the suppression of TPA (12-O-tetradecanoylphorbol-13-acetate)-induced inflammation. These results suggest that cold pressed unrefined canola/rapeseed oil, or other oils with high levels of lutein and other carotenoids, may be useful for their anti-inflammatory properties.
• Brown Sea Weed Lipids as Possible Source for Nutraceuticals and Functional Foods

  M. Airanthi K. Widjaja-Adhi, Takayuki Tsukui, Masashi Hosokawa, Kazuo Miysahita

  Biocatalysis and Biomolecular Engineering 211 - 223 2010/07/19 [Refereed][Not invited]
  
• Suppressive effects of the marine carotenoids, fucoxanthin and fucoxanthinol on triglyceride absorption in lymph duct-cannulated rats

  Megumi Matsumoto, Masashi Hosokawa, Noriko Matsukawa, Masahito Hagio, Aki Shinoki, Megumi Nishimukai, Kazuo Miyashita, Takaji Yajima, Hiroshi Hara

  EUROPEAN JOURNAL OF NUTRITION 49 (4) 243 - 249 1436-6207 2010/06 [Refereed][Not invited]
   

  Fucoxanthin isolated from edible seaweeds and its metabolite fucoxanthinol have been recently found to have anti-obesity effects, but the mechanism is not fully understood.We investigated the effects of these carotenoids on the absorption of triglycerides in conscious rats implanted with cannulae into a lymph duct and the portal or jugular vein.A duodenal infusion of 1 ml of test oil emulsion with or without 2 mg of fucoxanthin or fucoxanthinol was administered in the lymph duct and the portal (Experiment 1) or the jugular vein (Experiment 2) cannulated rats. The test oil contained 10% soybean oil (Experiment 1) and pre-digested 10% soybean oil (Experiment 2). The inhibitory activities of these carotenoids on pancreatic lipase activity were measured in vitro.Increases in lymphatic and blood triglyceride levels were much lower in the two carotenoid-treated groups than in the carotenoid-free group, indicating that these carotenoids inhibit triglyceride absorption. The total amounts of triglycerides released into the lymph after 4 h in the carotenoid-free, fucoxanthin and fucoxanthinol groups were 113.5, 59.4 and 53.1 mu mol, respectively. The inhibitory effects of carotenoids were completely abolished after an infusion of pre-digested soybean oil containing carotenoids. Furthermore, these carotenoids inhibited pancreatic lipase activity in vitro. Regarding absorptive route, we found that fucoxanthinol, but not fucoxanthin, appeared in lymph fluid, whereas neither carotenoid was detected in portal blood.These results show that these two marine carotenoids inhibit lipase activity in the gastrointestinal lumen and suppress triglyceride absorption, and fucoxanthin was converted to fucoxanthinol in the intestine and released into the lymph.
• Effects of sea squirt (Halocynthia roretzi) lipids on white adipose tissue weight and blood glucose in diabetic/obese KK-A(y) mice

  Nana Mikami, Masashi Hosokawa, Kazuo Miyashita

  MOLECULAR MEDICINE REPORTS 3 (3) 449 - 453 1791-2997 2010/05 [Refereed][Not invited]
   

  Lipids extracted from Halocynthia roretzi contain n-3 polyunsaturated fatty acids such as eicosapentaenoic acid and docosahexaenoic acid, as well as carotenoids. The aim of the present study was to evaluate the effect of H. roretzi lipids on white adipose tissue (WAT) weight and high blood glucose levels in diabetic/obese KK-A(y) mice. H. roretzi lipids were fed to the diabetic/obese KK-A(y) mice for 5 weeks. In the mice treated with the H. roretzi lipids compared to control mice, WAT weight was reduced, blood glucose levels and leptin mRNA expression in the epididymal WAT were significantly decreased, serum leptin levels also tended to decrease, and serum adiponectin levels tended to increase. These results demonstrate that H. roretzi lipids have beneficial health effects on diabetic/obese KK-A(y) mice.
• Radical scavenging and singlet oxygen quenching activity of extracts from Indian seaweeds

  N. M. Sachindra, M. K. W. A. Airanthi, M. Hosokawa, K. Miyashita

  JOURNAL OF FOOD SCIENCE AND TECHNOLOGY-MYSORE 47 (1) 94 - 99 0022-1155 2010/01 [Refereed][Not invited]
   

  Free radicals and singlet oxygen are responsible for oxidative stress related diseases and many natural compounds are known to have antioxidant properties. In this study, extracts from brown and red seaweeds of Indian origin were evaluated for their ability to scavenge different radicals and quench singlet oxygen. The crude extract in methanol and its fractions in different solvents were evaluated for their activity. The methanol extract and its fractions from brown seaweed exhibited higher 2,2'-azinobis(3-ethyl-benzothizoline-6-sulfonic acid) radical scavenging activity with more than 90% scavenging in butanol and ethyl acetate fractions and correlated with polyphenol content. There was a significant difference (p <= 0.001) in hydroxyl radical scavenging activity between different fractions of the same seaweed. Among the crude extracts, extract from Gracilaria corticata showed the highest (14.0%) activity. Crude extract from brown seaweeds showed higher peroxyl radical scavenging activity compared to red seaweeds. In fractions from brown seaweed extracts, highest activity was observed in ethyl acetate fraction (>88%) followed by hexane fraction (>40%). Ethyl acetate fraction from crude extract showed higher inhibitory activity against hemoglobin induced linoleic acid oxidation. Singlet oxygen quenching activity of the crude extract from brown seaweed was lower (<13%) compared to red seaweeds (16.4-20.5%).
• In vitro and in vivo evaluation of mutagenicity of fucoxanthin (FX) and its metabolite fucoxanthinol (FXOH)

  Fumiaki Beppu, Yoshimi Niwano, Emiko Sato, Masahiro Kohno, Takayuki Tsukui, Masashi Hosokawa, Kazuo Miyashita

  JOURNAL OF TOXICOLOGICAL SCIENCES 34 (6) 693 - 698 0388-1350 2009/12 [Refereed][Not invited]
   

  Mutagenicity of fucoxanthinol (FXOH), the major compound after oral ingestion of fucoxanthin (FX), was evaluated by in vitro Ames test, and of FX by in vivo micronucleus test. In in vitro Ames test, bacterial reverse mutation was examined by using Salmonella typhimurium strains TA98, TA100. TA1535 and TA1537, and Escherichia coli Wp2uvrA/pKM101, with or without metabolic activation by S9 mix in the preincubation method, and mutagenicity of FXOH was found to be negative in all cases. In in vivo micronucleus test, mice were orally administered with FX at doses of 500, 1,000 and 2,000 mg/kg, and the bone marrow cells were taken 24 hr after the administration to observe the incidence of micronucleus cells, and mutagenicity of FX was found to be negative at all doses. Based on the data of the present Study it can be presumed that orally administered FX is a safe compound in terms of mutagenicity under the experimental conditions employed here.
• Inhibition of proliferation of a hepatoma cell line by fucoxanthin in relation to cell cycle arrest and enhanced gap junctional intercellular communication

  Cheng-Ling Liu, Yung-Sheng Huang, Masashi Hosokawa, Kazuo Miyashita, Miao-Lin Hu

  CHEMICO-BIOLOGICAL INTERACTIONS 182 (2-3) 165 - 172 0009-2797 2009/12 [Refereed][Not invited]
   

  Fucoxanthin is one of the most abundant carotenoids found in Undaria pinnatifida and has been shown to inhibit tumor proliferation in vitro. However, the mechanisms underlying the anti-cancer effects of fucoxanthin are unclear. In this study, we hypothesized that fucoxanthin may cause cell cycle arrest and enhance gap junctional intercellular communication (GJIC) in SK-Hep-1 human hepatoma cells. Data revealed that fucoxanthin (1-20 mu M) strongly and concentration-dependently inhibited the proliferation of SK-Hep-1 cells at 24 h of incubation, whereas fucoxanthin facilitated the growth of a murine embryonic hepatic (BNL CL2) cells at 24 h of incubation and only slightly slowed the cell proliferation at 48 h. In SK-Hep-1 cells, fucoxanthin caused cell cycle arrest at G0/G1 phase and induced cell apoptosis, as evidenced by increased subG1 cells and induction of DNA strand breaks. Using scrape loading-dye-transfer assay, fucoxanthin was found to significantly enhance GJIC of SK-Hep-1 cells without affecting that of BNL CL2 cells. In addition, fucoxanthin significantly increased protein and mRNA expressions of connexin 43 (Cx43) and connexin 32 (Cx32) in SK-Hep-1 cells. Moreover, fucoxanthin markedly increased the concentration of intracellular calcium levels in SK-Hep-1 cells. Thus, fucoxanthin is specifically antiproliferative against SK-Hep-1 cells, and the effect is associated with upregulation of Cx32 and Cx43, which enhances GJIC of SK-Hep-1 cells. The enhanced GJIC may be responsible for the increase of the intracellular calcium level, which then causes cell cycle arrest and apoptosis. (C) 2009 Elsevier Ireland Ltd. All rights reserved.
• Effect of Marine Protein Supplementation on Lipid Profile of Growing Rats Compared to Soybean Protein and Casein

  Bhasakar Narayan, Kohei Yamaguchi, Masashi Hosokawa, Kenji Fukunaga, Toshimasa Nishiyama, Kazuo Miyashita

  FOOD SCIENCE AND BIOTECHNOLOGY 18 (6) 1330 - 1335 1226-7708 2009/12 [Refereed][Not invited]
   

  Although beneficial effects of dietary plant proteins on lipid metabolism are well documented, not much information exists on the influence of different seafood proteins on the lipid metabolism. The present study evaluated the effect of 2 marine proteins (tuna protein and scallop ovary proteins) in comparison to casein and soy protein in male Wistar rats. The concentration of total lipids in the plasma of rats fed experimental diets was significantly lower from that of control (278.2 mg/dL) group (p<0.05); and, the liver lipid content was not significantly different (p>0.05). Fecal excretion of cholesterol and bile acids was significantly higher in marine proteins and soy protein fed groups compared to casein only fed control (6.1 and 6.4 mg/day, respectively) group (p<0.05). No significant differences were observed in the mRNA concentrations of different transcriptional factors (p>0.05).
• Anti-obesity and anti-diabetic effects of fucoxanthin on diet-induced obesity conditions in a murine model

  Hayato Maeda, Masashi Hosokawa, Tokutake Sashima, Katsura Murakami-Funayama, Kazuo Miyashita

  MOLECULAR MEDICINE REPORTS 2 (6) 897 - 902 1791-2997 2009/11 [Refereed][Not invited]
   

  Fucoxanthin, a characteristic carotenoid of brown algae, has been reported to exert an anti-diabetic effect in an obese murine model. Wakame (Undaria pinnatifida), an edible seaweed, is rich in fucoxanthin. This study examined the anti-obesity and anti-diabetic effects of fucoxanthin-rich wakame lipids (WLs) on high fat (HF) diet-induced obesity in mice. Mice were fed a high fat control (HFC) or normal fat control (NFC) diet for 10 weeks. The HF diet-fed group was administered a HF diet containing WLs for a further 5 weeks. Parameters related to diabetes and obesity conditions were evaluated and compared. The HF-WL diet, which was rich in fucoxanthin, significantly suppressed body weight and white adipose tissue (WAT) weight gain induced by the HF diet. Dietary administration of the HF diet resulted in hyperglycemia, hyperinsulinemia and hyperleptinemia in the mouse model. These perturbations were completely normalized in the HF-WL diet-fed group. Increased expression of monocyte chemoattractant protein-1 (MCP-1) mRNA expression was observed in HFC mice, but was normalized in the HF-WL groups. Moreover, the HF-WL diet promoted mRNA expression of beta 3-adrenergic receptor (Adrb3) in WAT and glucose transporter 4 (GLUT4) mRNA in skeletal muscle tissues. These results suggest that dietary WLs may ameliorate alterations in lipid metabolism and insulin resistance induced by a HF diet. There is therefore a biochemical and nutritional basis for the application of fucoxanthin-rich WLs as a functional food to prevent obesity and diabetes-related disorders.
• Single and repeated oral dose toxicity study of fucoxanthin (FX), a marine carotenoid, in mice

  Fumiaki Beppu, Yoshimi Niwano, Takayuki Tsukui, Masashi Hosokawa, Kazuo Miyashita

  JOURNAL OF TOXICOLOGICAL SCIENCES 34 (5) 501 - 510 0388-1350 2009/10 [Refereed][Not invited]
   

  Fucoxanthin (FX), a xanthophyll derivative, is ail orange-colored pigment present in edible brown algae. As a part of safety evaluation, single and repeated oral dose toxicity study of FX was conducted. In a single dose study, FX purified from seaweed was orally administered to male and female ICR mice at doses of 1,000 and 2,000 mg/kg. In a repeated doses study, FX at doses of 500 and 1,000 mg/kg was orally administered for 30 days. In both studies, no mortality and no abnormalities in gross appearance were observed. In the repeated doses Study, histological observation revealed no abnormal changes in liver, kidney, spleen and gonadal tissues of any of the FX-treated groups. However, significantly increased total cholesterol concentrations were shown by plasma biochemical analyses in all FX-treated groups. Although total bilirubin concentrations were increased by FX, it was established that presence of fucoxanthinol, a major metabolite of FX, interfered with bilirubin determination in plasma. To further ascertain the safety of FX, the mechanism by which FX induces hypercholesterolemia in mice and species differences in the induction of hypercholesterolemia should be elucidated.
• EVALUATION OF RECOVERABLE FUNCTIONAL LIPID COMPONENTS OF SEVERAL BROWN SEAWEEDS (PHAEOPHYTA) FROM JAPAN WITH SPECIAL REFERENCE TO FUCOXANTHIN AND FUCOSTEROL CONTENTS

  Masaru Terasaki, Atsushi Hirose, Bhaskar Narayan, Yuta Baba, Chikara Kawagoe, Hajime Yasui, Naotsune Saga, Masashi Hosokawa, Kazuo Miyashita

  JOURNAL OF PHYCOLOGY 45 (4) 974 - 980 0022-3646 2009/08 [Refereed][Not invited]
   

  Fucoxanthin (Fx) and fucosterol (Fs) are characteristic lipid components of brown seaweeds that afford several health benefits to humans. This article describes the quantitative evaluation of lipids of 15 species of brown seaweeds with specific reference to Fx, Fs, and functional long-chain omega-6/omega-3 polyunsaturated fatty acids (PUFAs). In addition, fatty-acid composition of selected species was also accomplished in the study. Major omega-3 PUFAs in the brown seaweeds analyzed were alpha-linolenic acid (18:3n-3), octadecatetraenoic acid (18:4n-3), arachidonic acid (20:4n-6), and eicosapentaenoic acid (20:5n-3). Both Fx (mg center dot g(-1) dry weight [dwt]) and Fs (mg center dot g(-1) dwt) were determined to be relatively abundant in Sargassum horneri (Turner) C. Agardh (Fx, 3.7 +/- 1.6; Fs, 13.4 +/- 4.4) and Cystoseira hakodatensis (Yendo) Fensholt (Fx, 2.4 +/- 0.9; Fs, 8.9 +/- 2.0), as compared with other brown seaweed species. Studies related to seasonal variation in Fx, Fs, and total lipids of six brown algae [S. horneri, C. hakodatensis, Sargassum fusiforme (Harv.) Setch., Sargassum thunbergii (Mertens ex Roth) Kuntze, Analipus japonicus (Harv.) M. J. Wynne, and Melanosiphon intestinalis (D. A. Saunders) M. J. Wynne] indicated that these functional lipid components reached maximum during the period between January and March. The functional lipid components present in these seaweeds have the potential for application as nutraceuticals and novel functional ingredients after their recovery.
• Edible Japanese seaweed, wakame (Undaria pinnatifida) as an ingredient in pasta: Chemical, functional and structural evaluation

  P. Prabhasankar, P. Ganesan, N. Bhaskar, A. Hirose, Nimishmol Stephen, Lalitha R. Gowda, M. Hosokawa, K. Miyashita

  FOOD CHEMISTRY 115 (2) 501 - 508 0308-8146 2009/07 [Refereed][Not invited]
   

  Wakame (Undaria pinnatifida) is edible seaweed rich in fucoxanthin; whilst, pasta is an important dish from nutritional and gastronomic point of view. Pasta was prepared with wakame as an ingredient at different levels. In vitro antioxidant properties, total phenolic content, fatty acid composition, fucoxanthin and fucosterol contents formed the major bio-functional characteristics analysed. Pasta with 10% wakame was acceptable sensorially. The total phenolic content varied between 0.10 and 0.94 mg gallic acid equivalents (GAE)/g, whilst total antioxidant activity varied from 0.16 to 2.14 mg ascorbic acid equivalents (AAE)/g, amongst different samples. 2,2-Diphenyl-1-picrylhydrazyl (DPPH) and superoxide radical scavenging activities of sensorily acceptable pasta were 7.71% and 4.56%, respectively. The sensorily acceptable pasta had a mild seaweed flavour with taste similar to control pasta, as assessed by panelists. The ratio of n-3 to n-6 fatty acid in seaweed incorporated pasta was 1:3.4 as compared to 1:15.2 in the control. Heat process involved in pasta preparation and cooking did not destroy fucoxanthin. Microstructure studies revealed the enhanced interaction between starch granules and protein matrix in pasta containing seaweeds up to 20%. (C) 2008 Elsevier Ltd. All rights reserved.
• Antiobesity Effect of Allenic Carotenoid, Fucoxanthin

  Kazuo Miyashita, Masashi Hosokawa

  Nutrigenomics and Proteomics in Health and Disease: Food Factors and Gene Interactions 145 - 160 2009/06/12 [Not refereed][Not invited]
  
• Effect of Droplet Size on the Oxidative Stability of Soybean Oil TAG and Fish Oil TAG in Oil-in-Water Emulsion

  Goki Azuma, Naoko Kimura, Masashi Hosokawa, Kazuo Miyashita

  JOURNAL OF OLEO SCIENCE 58 (6) 329 - 338 1345-8957 2009/06 

  The effect of droplet size on the oxidative stability of triacylglycerol (TAG)-in water emulsion was examined. Microchannel (MC) emulsification was used to make monodispersed emulsion of soybean oil TAG and fish oil TAG with different droplet size. The main polyunsaturated fatty acids (PUFA) of soybean oil TAG and fish oil TAG were linoleic acid (LA; 53%) and docosahexaenoic acid (DHA; 37.3%), respectively. Oxidation was induced by the addition of 2,2'-azobis(2-amidinopropane)-dihydrochloride (AAPH) or ferrous ion. The oxidative stability was followed by the decrease in the oxygen consumption in the solution, peroxide formation, and decrease in the unoxidized PUFA during oxidation, indicating that the oxidative stability of fish oil TAG increased with decreasing the droplet size, while the reverse effect of the droplet size was observed on the oxidation of soybean oil TAG. The decrease in the droplet size induces the increase in the droplet interface, from which the oxidation proceeds to the oil droplet interior. DHA in fish oil TAG would take highly protective interface against oxidative attack of free radicals and metal ions, whereas LA in soybean oil TAG would be more easily oxidized at the interface because of its less protective conformation. The reverse effect of the droplet size on fish oil TAG and soybean oil TAG could be explained by the different interface conformation of both TAG.
• Effect of Droplet Size on the Oxidative Stability of Soybean Oil TAG and Fish Oil TAG in Oil-in-Water Emulsion

  Goki Azuma, Naoko Kimura, Masashi Hosokawa, Kazuo Miyashita

  JOURNAL OF OLEO SCIENCE 58 (6) 329 - 338 1345-8957 2009/06 [Refereed][Not invited]
   

  The effect of droplet size on the oxidative stability of triacylglycerol (TAG)-in water emulsion was examined. Microchannel (MC) emulsification was used to make monodispersed emulsion of soybean oil TAG and fish oil TAG with different droplet size. The main polyunsaturated fatty acids (PUFA) of soybean oil TAG and fish oil TAG were linoleic acid (LA; 53%) and docosahexaenoic acid (DHA; 37.3%), respectively. Oxidation was induced by the addition of 2,2'-azobis(2-amidinopropane)-dihydrochloride (AAPH) or ferrous ion. The oxidative stability was followed by the decrease in the oxygen consumption in the solution, peroxide formation, and decrease in the unoxidized PUFA during oxidation, indicating that the oxidative stability of fish oil TAG increased with decreasing the droplet size, while the reverse effect of the droplet size was observed on the oxidation of soybean oil TAG. The decrease in the droplet size induces the increase in the droplet interface, from which the oxidation proceeds to the oil droplet interior. DHA in fish oil TAG would take highly protective interface against oxidative attack of free radicals and metal ions, whereas LA in soybean oil TAG would be more easily oxidized at the interface because of its less protective conformation. The reverse effect of the droplet size on fish oil TAG and soybean oil TAG could be explained by the different interface conformation of both TAG.
• Effect of alpha-tocopherol on the synthesis of phosphatidylglycerol catalyzed by phospholipase D in an aqueous system

  Kounosuke Suzuri, Yukihiro Yamamoto, Masashi Hosokawa, Kazuo Miyashita

  BIOTECHNOLOGY LETTERS 31 (5) 719 - 723 0141-5492 2009/05 [Refereed][Not invited]
   

  Phosphatidylglycerol (PG) was synthesized from several phosphatidylcholines (PCs) via phospholipase D (PLD)-catalyzed transphosphatidylation in an aqueous system. The yield of PG were 71 and 68 mol% from soybean PC and egg yolk PC, respectively, under the optimum reaction conditions of 50 mu mol PC, 10 mmol glycerol, 3 ml of acetate buffer, 1.6 U PLD, and 30 mu mol CaCl2 at 37A degrees C for 48 h. In case of salmon roe PC with 14.3% eicosapentaenoic acid and 26.8% docosahexaenoic acid, the PG yield increased to 94 mol% by addition of 46 mu mol alpha-tocopherol, although the PG yield was only 10% in absence of alpha-tocopherol.
• 褐藻由来フコキサンチンのマウス肝臓におけるΔ6-不飽和化酵素タンパク質の発現増加作用

  津久井 隆行, 細川 雅史, 宮下 和夫

  日本栄養・食糧学会大会講演要旨集 (公社)日本栄養・食糧学会 63回 234 - 234 2009/05
• Enhancement of hepatic docosahexaenoic acid and arachidonic acid contents in C57BL/6J mice by dietary fucoxanthin

  Takayuki Tsukui, Nobuko Baba, Masashi Hosokawa, Tokutake Sashima, Kazuo Miyashita

  FISHERIES SCIENCE 75 (1) 261 - 263 0919-9268 2009/02 [Refereed][Not invited]
  
• Bio-functions of Marine Carotenoids

  Masashi Hosokawa, Tomoko Okada, Nana Mikami, Izumi Konishi, Kazuo Miyashita

  FOOD SCIENCE AND BIOTECHNOLOGY 18 (1) 1 - 11 1226-7708 2009/02 [Refereed][Not invited]
   

  Carotenoids being most important pigments among those occurring in nature, have received increased interest owing to their beneficial effects on human health. An effort is made to review marine carotenoids as important bioactive compounds with reference to their presence, chemical, and biofunctional benefits they afford. The potential beneficial effects of marine carotenoids were particularly focused on astaxanthin and fucoxanthin, major marine carotenoids found in marine animals and aquatic plants, respectively. Both carotenoids show strong antioxidant activity which is attributed to quenching singlet oxygen and scavenging free radicals. The potential role of the carotenoids as dietary antioxidants has been suggested as being one of the main mechanism by which they afford their beneficial health effects such as anticancer activity and anti-inflammatory effect. Only recently, antiobesity effect and antidiabetic effect have been noted as specific and novel biofunctions of fucoxanthin. Nutrigenomic study reveals that fucoxanthin induces uncoupling protein 1 (UCP1) expression in white adipose tissue (WAT) mitochondria to lead to oxidation of fatty acids and heat production in WAT. Fucoxanthin improves insulin resistance and decreases blood glucose level, at least in part, through the down-regulation of tumor necrosis factor alpha (TNF alpha) in WAT of animals.
• Comparative evaluation of growth inhibitory effect of stereoisomers of fucoxanthin in human cancer cell lines

  Y. Nakazawa, T. Sashima, M. Hosokawa, K. Miyashita

  JOURNAL OF FUNCTIONAL FOODS 1 (1) 88 - 97 1756-4646 2009/01 [Refereed][Not invited]
   

  Inhibitory effects of geometrical isomers of fucoxanthin - characteristic carotenoid of brown seaweeds - on the growth of various cancer cells and on human leukemia (HL-60), colon cancer (Caco-2) and prostate cancer cells (PC-3 and LNCap) were comparatively evaluated. All-trans fucoxanthin was the major geometrical isomer (similar to 88%) found in the fresh brown seaweed (Undaria pinnatifida) apart from a small amount of 13-cis and 13'-cis isomers (similar to 9%). Incubation of the fucoxanthin isomeric mixtures, all-trans fucoxanthin with a small amount of 13-cis and 13'-cis isomers, produced 9'-cis isomer (5%) and increased the contents of 13-cis and 13'-cis isomers (27%). The antiproliferative effect of the mixture of 13-cis and 13'-cis isomers was stronger than all other geometrical isomers evaluated in the study. The inhibition of growth of HL-60 cells was higher in case of 13'-cis isomer followed by 13-cis and all-trans isomers. The potent inhibitory effect of 13-cis and 13'-cis fucoxanthin on HL-60 cells and Caco-2 cells could possibly be due to their higher apoptosis-inducing activity. (C) 2008 Elsevier Ltd. All rights reserved.
• Eicosapentaenoic Acid Plays a Beneficial Role in Membrane Organization and Cell Division of a Cold-Adapted Bacterium, Shewanella livingstonensis Ac10

  Jun Kawamoto, Tatsuo Kurihara, Kentaro Yamamoto, Makiko Nagayasu, Yasushi Tani, Hisaaki Mihara, Masashi Hosokawa, Takeshi Baba, Satoshi B. Sato, Nobuyoshi Esaki

  JOURNAL OF BACTERIOLOGY 191 (2) 632 - 640 0021-9193 2009/01 [Refereed][Not invited]
   

  Shewanella livingstonensis Ac10, a psychrotrophic gram-negative bacterium isolated from Antarctic seawater, produces eicosapentaenoic acid (EPA) as a component of phospholipids at low temperatures. EPA constitutes about 5% of the total fatty acids of cells grown at 4 degrees C. We found that five genes, termed orf2, orf5, orf6, orf7, and orf8, are specifically required for the synthesis of EPA by targeted disruption of the respective genes. The mutants lacking EPA showed significant growth retardation at 4 degrees C but not at 18 degrees C. Supplementation of a synthetic phosphatidylethanolamine that contained EPA at the sn-2 position complemented the growth defect. The EPA-less mutant became filamentous, and multiple nucleoids were observed in a single cell at 4 degrees C, indicating that the mutant has a defect in cell division. Electron microscopy of the cells by high-pressure freezing and freeze-substitution revealed abnormal intracellular membranes in the EPA-less mutant at 4 degrees C. We also found that the amounts of several membrane proteins were affected by the depletion of EPA. While polyunsaturated fatty acids are often considered to increase the fluidity of the hydrophobic membrane core, diffusion of a small hydrophobic molecule, pyrene, in the cell membranes and large unilamellar vesicles prepared from the lipid extracts was very similar between the EPA-less mutant and the parental strain. These results suggest that EPA in S. livingstonensis Ac10 is not required for bulk bilayer fluidity but plays a beneficial role in membrane organization and cell division at low temperatures, possibly through specific interaction between EPA and proteins involved in these cellular processes.
• Anti-obesity and anti-diabetic effects of fucoxanthin on diet-induced obesity conditions in a murine model

  Hayato Maeda, Masashi Hosokawa, Tokutake Sashima, Katsura Murakami-Funayama, Kazuo Miyashita

  MOLECULAR MEDICINE REPORTS 2 (6) 897 - 902 1791-2997 2009 [Refereed][Not invited]
   

  Fucoxanthin, a characteristic carotenoid of brown algae, has been reported to exert an anti-diabetic effect in an obese murine model. Wakame (Undaria pinnatifida), an edible seaweed, is rich in fucoxanthin. This study examined the anti-obesity and anti-diabetic effects of fucoxanthin-rich wakame lipids (WLs) on high fat (HF) diet-induced obesity in mice. Mice were fed a high fat control (HF(C)) or normal fat control (NF(C)) diet for 10 weeks. The HF diet-fed group was administered a HF diet containing WLs for a further 5 weeks. Parameters related to diabetes and obesity conditions were evaluated and compared. The HF-WL diet, which was rich in fucoxanthin, significantly suppressed body weight and white adipose tissue (WAT) weight gain induced by the HF diet. Dietary administration of the HF diet resulted in hyperglycemia, hyperinsulinemia and hyperleptinemia in the mouse model. These perturbations were completely normalized in the HF-WL diet-fed group. Increased expression of monocyte chemoattractant protein-1 (MCP-1) mRNA expression was observed in HF(C) mice, but was normalized in the HF-WL groups. Moreover, the HF-WL diet promoted mRNA expression of beta 3-adrenergic receptor (Adrb3) in WAT and glucose transporter 4 (GLUT4) mRNA in skeletal muscle tissues. These results suggest that dietary WLs may ameliorate alterations in lipid metabolism and insulin resistance induced by a HF diet. There is therefore a biochemical and nutritional basis for the application of fucoxanthin-rich WLs as a functional food to prevent obesity and diabetes-related disorders.
• Anti-Obesity Effect of Allene Carotenoids, Fucoxanthin and Neoxanthin from Seaweeds and Vegetables

  K. Miyashita, H. Maeda, T. Tsukui, T. Okada, M. Hosokawa

  II INTERNATIONAL SYMPOSIUM ON HUMAN HEALTH EFFECTS OF FRUITS AND VEGETABLES: FAVHEALTH 2007 841 167 - 171 0567-7572 2009 [Not refereed][Not invited]
   

  Obesity is one of the most prevalent nutritional disorders, and it results from an imbalance between energy intake and expenditure. We have found that carotenoids containing an allene bond, such as fucoxanthin and fucoxanthinol, showed anti-obesity effects with a new molecular mechanism. We showed significant reduction of WAT in wistar rats and obese KK-Ay mice by feeding both carotenoids. Clear signals of UCP1 protein and mRNA were detected by Western and Northern blot analyses in WAT in mice fed fucoxanthin, although there is little expression of UCP1 in WAT in mice fed the control diet. UCP1 expression in WAT by fucoxanthin intake leads to oxidation of fatty acids and heat production in WAT mitochondria. The substrate oxidation can directly reduce WAT in animals. We also found a relationship between carotenoid structure and anti-obesity properties, i.e., ability to suppress adipocyte differentiation. An examination of structure and function revealed that carotenoids containing an allene bond and an additional hydroxyl substituent on the side group show suppressive effects on adipocyte differentiation in 3T3-L1 cells. These findings show a great potential for application in the field of therapeutics for the development of novel anti-obesity compounds. Further, the suggestion that carotenoid health effects are likely linked to structural characteristics may help in the discovery of additional carotenoids that possess health-promoting properties.
• ANTI-OBESITY AND ANTI-DIABETIC EFFECT OF ALLENIC CAROTENOID

  Tomoko Okada, Hayato Maeda, Masashi Hosokawa, Kazuo Miyashita

  ANNALS OF NUTRITION AND METABOLISM 55 92 - 92 0250-6807 2009 [Refereed][Not invited]
  
• Growth Inhibition and Induction of Apoptosis of Colon Cancer Cell Lines by Applying Marine Phospholipid

  Zakir Hossain, Masashi Hosokawa, Koretaro Takahashi

  NUTRITION AND CANCER-AN INTERNATIONAL JOURNAL 61 (1) 123 - 130 0163-5581 2009 [Refereed][Not invited]
   

  Polyunsaturated fatty acids (PUFAs) exhibit beneficial biological functions in carcinogenic processes. We examined the effects of PUFAs in the acid and phospholipid forms on three colon cancer cell lines (HT-29, Caco-2, and DLD-1). Docosahexaenoic acid (DHA) and eicosapentaenoic (EPA) in both acid and phospholipid forms showed growth inhibition effects on experimental colon cancer cell lines. But these PUFAs had the strongest growth-inhibitory effect on HT-29 than Caco-2 and DLD-1. Combined application of PUFAs and sodium butyrate (NaBt) increased the growth inhibition. Growth inhibition was apparently caused by increased lipid peroxidation. DHA or EPA in combination with NaBt significantly increased caspase-3 activity compared to control. DHA and DHA-rich phosphatidylcholine decreased Bcl-2 level in HT-29 and Caco-2 cells.
• New C-37 Skeletal Carotenoid from the Clam, Paphia amabillis

  Takashi Maoka, Naoshige Akimoto, Mi-Jin Yim, Masashi Hosokawa, Kazuo Miyashita

  JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY 56 (24) 12069 - 12072 0021-8561 2008/12 [Refereed][Not invited]
   

  Carotenoids in the muscle of Paphia amabillis (Veneridae) were investigated. Amarouciaxanthin 3-ester (11 a) was found to be a major carotenoid along with amarouciaxanthin A (1b), fucoxanthinol 3-ester (2), peridininol 3'-ester (3), and a new C-37 skeletal carotenoid, named hydratopyrrhoxanthinol Tester. This structure was determined to be 3'-acyloxy-3,5,6-trihydroxy-7',8'-didehydro-5,6-dihydro-12',13',20'-trinor-beta,beta-caroten-19,11-olide (4) by UV-vis, NMR, and FAB-MS spectral data. The absolute configuration of 4 was postulated to be (3S,5R,6R,3'R) on the bases of CD and ROESY data. The fatty acids esterified with hydratopyrrhoxanthinol were assigned as C22:6, C20:1, C20:4, C20:5, C18:0, C18:1, C17:0, C16:0, C16:1, and C14:0 on the basis of FAB-MS data.
• Cold adaptation of eicosapentaenoic acid-less mutant of Shewanella livingstonensis Ac10 involving uptake and remodeling of synthetic phospholipids containing various polyunsaturated fatty acids

  Sho Sato, Tatsuo Kurihara, Jun Kawamoto, Masashi Hosokawa, Satoshi B. Sato, Nobuyoshi Esaki

  EXTREMOPHILES 12 (6) 753 - 761 1431-0651 2008/11 [Refereed][Not invited]
   

  An Antarctic psychrotrophic bacterium, Shewanella livingstonensis Ac10, produces cis-5,8,11,14,17-eicosapentaenoic acid (EPA), a long-chain polyunsaturated fatty acid (LPUFA), as a component of membrane phospholipids at low temperatures. The EPA-less mutant generated by disruption of the EPA synthesis gene becomes cold-sensitive. We studied whether the cold sensitivity could be suppressed by supplementation of various LPUFAs. The EPA-less mutant was cultured at 6 degrees C in the presence of synthetic phosphatidylethanolamines (PEs) that contained oleic acid at the sn-1 position and various C20 fatty acids with different numbers of double bonds from zero to five or cis-4,7,10,13,16,19-docosahexaenoic acid (DHA) at the sn-2 position. Mass spectrometric analyses revealed that all these fatty acids became components of various PE and phosphatidylglycerol species together with shorter partner fatty acids, indicating that large-scale remodeling followed the incorporation of synthetic PEs. As the number of double bonds in the sn-2 acyl chain decreased, the growth rate decreased and the cells became filamentous. The growth was restored to the wild-type level only when the medium was supplemented with phospholipids containing EPA or DHA. We found that about a half of DHA was converted into EPA. The results suggest that intact EPA is best required for cold adaptation of this bacterium.
• Effects of Trans and Conjugated LC N-3 Polyunsaturated Fatty Acids on Lipid Composition and Abdominal Fat Weight in Rats

  T. Okada, R. Noguchi, M. Hosokawa, K. Fukunaga, T. Nishiyama, N. Zaima, T. Hirata, K. Miyashita

  JOURNAL OF FOOD SCIENCE 73 (8) H201 - H206 0022-1147 2008/10 [Refereed][Not invited]
   

  Trans and conjugated fatty acids may exhibit either beneficial or detrimental bioactive effects depending on their metabolic properties. This study was conducted to elucidate if isomerization and conjugation of eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) demonstrate more favorable bioactivity on lipid metabolism compared to unmodified EPA and DHA. The effects of dietary intake of trans and conjugated forms of EPA and DHA on lipid metabolism were evaluated in animal trials and compared to a control group fed soybean oil. None of the experimental diets showed significant differences from the control in terms of body weight; however, the white adipose tissue weight of rodents fed trans DHA, conjugated EPA (CEPA), and conjugated DHA (CDHA) was significantly lower than the control. Triacylglycerol levels in plasma were significantly decreased in groups fed trans DHA (17.2 mg/dL) and CDHA (31.9 mg/dL) relative to the control (51.3 mg/dL). The total cholesterol concentrations were significantly lower than the control (68.0 mg/dL) in all experimental groups (47.3 to 53.7 mg/dL) except CEPA (58.3 mg/dL). Fatty acid compositions of lipids extracted from rodent livers were influenced by the dietary fatty acid profiles, with all groups showing higher concentrations of stearic acid and lower levels of linoleic acid compared to the control. Rodents fed trans DHA did not have detectable levels of these fatty acid isomers in their livers, suggesting either quick metabolism or a difficulty with bio-absorption.
• Antiobesity effect of fucoxanthin from edible seaweeds and its multibiological functions

  Hayato Maeda, Masashi Hosokawa, Tokutake Sashima, Kazuo Miyashita

  ACS Symposium Series 993 376 - 388 1947-5918 2008/09/19 [Not refereed][Not invited]
   

  Fucoxanthin has a unique structure including an unusual allenic bond and 5,6-monoepoxide in its molecule. It induced a remarkable reduction in the growth of leukemia and prostate cancer cells. Furthermore, we have found that fucoxanthin showed anti-obesity effect with a new molecular mechanism. Uncoupling protein (UCP) is inner-membrane mitochondrial protein that has the ability to dissipate energy through uncoupling of oxidative phosphorylation which, instead of ATP, produces heat. A great deal of interest has focused on adaptive thermogenesis by UCP families (UCP1, 2 and 3) in several tissues and organs as a physiological defense against obesity, hyperlipidemia, and diabetes. In fact, UCP1 expression in brown adipose tissue (BAT) is known as a significant component of whole body energy expenditure, at least small rodents, and its dysfunction contributes to the development of obesity. However, adult humans have very little BAT, making it unlikely to be a major contributor to human weight regulation. In humans, most of fat is stored in white adipose tissue (WAT). Considered as breakthrough discoveries for an ideal therapy of obesity, regulation of UCP expression in white adipose tissue (WAT) by food constituent should be studied. Here, we show significant reduction of WAT in wistar rats and obese KK-Ay mice by feeding fucoxanthin (0.05 or 0.2 wt%/feed).Fucoxanthin, concentrated from edible seaweed, Undaria Pinnatifida (Japanese name is Wakame). The daily intake of fucoxanthin to KK-Ay mice also caused a significant reduction of body weight and a significant increase in BAT weight. Clear signals of UCP1 protein and mRNA were detected by Western and Northern blot analyses in WAT in mice fed fucoxanthin, although there is little expression of UCP1 in WAT in mice fed control diet. UCP1 expression in WAT by fucoxanthin intake leads to oxidation of fatty acids and heat production in WAT mitochondria. The substrate oxidation can directly reduce WAT in animals. © 2008 American Chemical Society.
• Synthesis of phosphatidylated-monoterpene alcohols catalyzed by phospholipase D and their antiproliferative effects on human cancer cells

  Yukihiro Yamamoto, Masashi Hosokawa, Hideyuki Kurihara, Takashi Maoka, Kazuo Miyashita

  BIOORGANIC & MEDICINAL CHEMISTRY LETTERS 18 (14) 4044 - 4046 0960-894X 2008/07 [Refereed][Not invited]
   

  In order to prepare functional phospholipids in the medical and pharmaceutical fields, perillyl alcohol, myrtenol, and nerol were transphosphatidylated via phospholipase D in an aqueous system. The yields of phosphatidyl-perillyl alcohol, -myrtenol, and -nerol were 79 mol %, 87 mol %, and 91 mol %, respectively. The synthetic phosphatidylated monoterpenes showed a markedly antiproliferative effect on human prostate PC-3 and human leukemia HL-60 cells at 100 mu M, while the free monoterpene alcohols had no effect at 400 mu M. (c) 2008 Elsevier Ltd. All rights reserved.
• Suppressive Effect of Neoxanthin on the Differentiation of 3T3-L1 Adipose Cells

  Tomoko Okada, Miho Nakai, Hayato Maeda, Masashi Hosokawa, Tokutake Sashima, Kazuo Miyashita

  JOURNAL OF OLEO SCIENCE 57 (6) 345 - 351 1345-8957 2008/06 [Refereed][Not invited]
   

  The objective of this study was to assess the suppressive effects of 13 naturally occurring carotenoids on the adipocyte differentiation of 3T3-L1. The relationship between carotenoid structure and suppressive effects was also examined. Treatment with neoxanthin significantly reduced lipid accumulation, as well as glycerol-3-phosphate dehydrogenase activity. This suppressive effect on adipose cell differentiation was not observed in the other 12 carotenoids used in this study. Neoxanthin treatment also decreased expression of CCAAT/enhancer binding protein alpha (C/EBP alpha) and peroxisome proliferator-activated receptor gamma (PPAR gamma) mRNAs. An examination of structure and function suggested that carotenoids containing an allene bond and an additional hydroxyl substituent on the side group may show suppressive effects on adipocyte differentiation in 3T3-L1 cells.
• Preparation of phosphatidylated terpenes via phospholipase D-mediated transphosphatidylation

  Yukihiro Yamamoto, Masashi Hosokawa, Hideyuki Kurihara, Kazuo Miyashita

  JOURNAL OF THE AMERICAN OIL CHEMISTS SOCIETY 85 (4) 313 - 320 0003-021X 2008/04 [Refereed][Not invited]
   

  Terpenes such as geraniol, geranylgeraniol, farnesol, and phytol are known as functional compounds which exhibit anticancer effects and activate nuclear receptors. For the application of functional terpenes in various fields, including the cosmetic and food industries, we attempted to synthesize phosphatidylated terpenes (terpene-PLs) by using phospholipase D (PLD). Transphosphatidylation of phosphatidylcholine with terpenes was carried out using PLD in a biphasic system containing ethyl acetate/water or in an aqueous system without organic solvent. The yield of terpene-PL increased with the reaction time and the amount of PLD in both the biphasic and aqueous systems. Further, the yield of terpene-PL in the aqueous system was higher than that in the biphasic system. In addition, among four PLDs from Streptomyces sp., Streptomyces chromofuscus, cabbage, and peanut, only the PLD from Streptomyces sp. could synthesize terpene-PL. The reaction yield, based on substrate phospholipid, of phosphatidylgeraniol reached 90 mol% under the following optimal reaction conditions: 50 mu mol soyPC; 2,000 mu mol geraniol; 1.6 U PLD; 0.8 ml of 0.2 M sodium acetate buffer (pH 5.6); temperature, 37 degrees C ; and reaction time, 24 h. The reaction yields of phosphatidylfarnesol, phosphatidylgeranylgeraniol, and phosphatidylphytol were 73, 54, and 17 mol%, respectively.
• Multi-functional carotenoid in seaweeds: fucoxanthin

  Kazuo Miyashita, Masashi Hosokawa

  NIPPON SUISAN GAKKAISHI 74 (2) 261 - 262 0021-5392 2008/03 [Refereed][Not invited]
  
• Suppressive Effects of Alloxanthin and Diatoxanthin from Halocynthia roretzi on LPS-induced Expression of Pro-inflammatory Genes in RAW264.7 Cells

  Izumi Konishi, Masashi Hosokawa, Tokutake Sashima, Takashi Maoka, Kazuo Miyashita

  JOURNAL OF OLEO SCIENCE 57 (3) 181 - 189 1345-8957 2008/03 [Refereed][Not invited]
   

  To investigate anti-inflammatory effects of carotenoids from Halocynthia roretzi, gene expression levels were measured for pro-inflammatory cytokines and enzymes in the murine macrophage-like cell line, RAW264.7, stimulated with lipopolysaccharide (LPS). All-trans alloxanthin, all-trans diatoxanthin, and their 9-cis isomers isolated from H. roretzi significantly suppressed expression of IL-1 beta and IL-6 mRNA in cells induced by LIPS without cytotoxicity. The expression level of IL-1 beta mRNA in cells treated with 25 mu M all-trans alloxanthin for 24 h, followed by stimulation with 0.1 mu g/mL LPS for 24 h in the presence of carotenoid decreased to 33.7 +/- 3.0% from that of control cells stimulated with LPS alone. All-trans diatoxanthin, 9-cis alloxanthin and 9-cis diatoxanthin also decreased expression of IL-1 beta mRNA to 25.1 +/- 2.1, 28.2 +/- 0.9 and 32.9 +/- 3.3%, respectively, from that of control cells stimulated with LPS. IL-1 beta production in culture medium was also suppressed by all-trans alloxanthin and all-trans diatoxanthin. Furthermore, all-trans alloxanthin, all-trans diatoxanthin and their 9-cis isomers suppressed the overexpression of cyclooxygenase-2 and nitric oxide synthase mRNA in RAW264.7 cells induced by LPS. The suppressive effects of these carotenoids were remarkable compared to those of beta-carotene and zeaxanthin.
• Enzymatic Production of Marine-Derived Protein Hydrolysates and Their Bioactive Peptides for Use in Foods and Nutraceuticals

  Tomoko Okada, Masashi Hosokawa, Seigo Ono, Kazuo Miyashita

  Biocatalysis and Bioenergy 491 - 519 2008/02/25 [Refereed][Not invited]
  
• Composition, Functionality and Potential Applications of Seaweed Lipids

  Bhaskar Narayan, Chandini S. Kumar, Tokutake Sashima, Hayato Maeda, Masashi Hosokawa, Kazuo Miyashita

  Biocatalysis and Bioenergy 463 - 490 2008/02/25 [Refereed][Not invited]
  
• Seaweed carotenoid, fucoxanthin, as a multi-functional nutrient

  Hayato Maeda, Takayuki Tsukui, Tokutake Sashima, Masashi Hosokawa, Kazuo Miyashita

  ASIA PACIFIC JOURNAL OF CLINICAL NUTRITION 17 196 - 199 0964-7058 2008 [Refereed][Not invited]
   

  Fucoxanthin has a unique structure including an unusual allenic bond and 5, 6-monoepoxide in its molecule. We found that abdominal white adipose tissue (WAT) weights of rats and mice fed fucoxanthin were significantly lower than those fed a control diet. The daily intake of fucoxanthin in mice also caused a significant reductions of body weight. Clear signals of uncoupling protein 1 (UCP1) and its mRNA were detected by Western and Northern blot analyses in abdominal WAT in mice fed fucoxanthin, although there is little expression of UCP1 in WAT in mice fed a control diet. UCP1 expression in WAT by fucoxanthin intake leads to oxidation of fatty acids and heat production in WAT mitochondria. Substrate oxidation can directly reduce WAT in animals. Fucoxanthin intake also significantly reduced blood glucose and plasma insulin. Furthermore, feeding fucoxanthin significantly increased the level of hepatic docosahexaenoic acid (DHA), a most important n-3 functional polyunsaturated fatty acid in biological systems. These multi-functionalities of fucoxanthin indicate that it is an important bioactive carotenoid that is should be beneficial for the prevention of the metabolic syndrome.
• Radical scavenging and singlet oxygen quenching activity of marine carotenoid fucoxanthin and its metabolites

  Nakkarike M. Sachindra, Emiko Sato, Hayato Maeda, Masashi Hosokawa, Yoshimi Niwano, Masahiro Kohno, Kazuo Miyashita

  JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY 55 (21) 8516 - 8522 0021-8561 2007/10 [Refereed][Not invited]
   

  Antioxidant activity of carotenoids is suggested to be one of the factors; for their disease preventing effects. Marine carotenoids fucoxanthin and its two metabolites, fucoxanthinol and halocynthiaxanthin, have been shown to exhibit several biological effects. The antioxidant activities of these three carotenoids were assessed in vitro with respect to radical scavenging and singlet oxygen quenching abilities. The 1,1 -diphenyl-2-picrylhydrazyl radical scavenging activity of fucoxanthin and fucoxanthinol was higher than that of halocynthiaxanthin, with the effective concentration for 50% scavenging (EC50) being 164.60, 153.78, and 826.39 mu M, respectively. 2,2'-Azinobis-3-ethylbenzo thizol ine-6-su I phonate radical scavenging activity of fucoxanthinol (EC50, 2.49 mu M) was stronger than that of fucoxanthin (EC50, 8.94 mu M). Hydroxyl radical scavenging activity as measured by the chemiluminescence technique showed that the scavenging activity of fucoxanthin was 7.9 times higher than that by fucoxanthinol, 16.3 times higher than that by halocynthiaxanthin, and 13.5 times higher than that by (x-tocopherol. A similar trend was observed when the hydroxyl radical scavenging was assessed by the electron spin resonance (ESR) technique. ESR analysis of the Superoxide radical scavenging activity also showed the superiority of fucoxanthin over the other two carotenoids tested. Singlet oxygen quenching ability of the three carotenoids was lower than that of beta-carotene, with quenching rate constants (k(Q), x 10(10) M-1 s(-1)) being 1.19, 1.81, 0.80, and 12.78 for fucoxanthin, fucoxanthinol, halocynthiaxanthin, and beta-carotene, respectively. The higher radical scavenging activity of fucoxanthin and fucoxanthinol compared with halocynthiaxanthin is assumed to be due to presence of the allenic bond.
• Dietary combination of fucoxanthin and fish oil attenuates the weight gain of white adipose tissue and decreases blood glucose in obese/diabetic KK-A(y) mice

  Hayato Maeda, Masashi Hosokawa, Tokutake Sashima, Kazuo Miyashita

  JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY 55 (19) 7701 - 7706 0021-8561 2007/09 [Refereed][Not invited]
   

  Fucoxanthin is a marine carotenoid found in edible brown seaweeds. We previously reported that dietary fucoxanthin attenuates the weight gain of white adipose tissue (WAT) of diabetic/obese KK-A(y) mice. In this study, to evaluate the antiobesity and antidiabetic effects of fucoxanthin and fish oil, we investigated the effect on the WAT weight, blood glucose, and insulin levels of KK-Ay mice. Furthermore, the expression level of uncoupling protein 1 (UCP1) and adipokine mRNA in WAT were measured. After 4 weeks of feeding, 0.2% fucoxanthin in the diet markedly attenuated the gain of WAT weight in KK-Ay mice with increasing UCP1 expression compared with the control mice. The WAT weight of the mice fed 0.1% fucoxanthin and 6.9% fish oil was also significantly lower than that of the mice fed fucoxanthin alone. In addition, 0.2% fucoxanthin markedly decreased the blood glucose and plasma insulin concentrations in KK-Ay mice. The mice fed with the combination diet of 0.1% fucoxanthin and fish oil also showed improvements similar to that of 0.2% fucoxanthin. Leptin and tumor necrosis factor (TNF alpha) mRNA expression in WAT were significantly down-regulated by 0.2% fucoxanthin. These results suggest that dietary fucoxanthin decreases the blood glucose and plasma insulin concentration of KK-Ay along with down-regulating TNFa mRNA. In addition, the combination of fucoxanthin and fish oil is more effective for attenuating the weight gain of WAT than feeding with fucoxanthin alone.
• Occurrence of conjugated cyclopropanoid acid in purified fish oil

  Naoto Horii, Shingo Arato, Bhaskar Narayan, Masashi Hosokawa, Tokutake Sashima, Kazuo Miyashita

  JOURNAL OF THE AMERICAN OIL CHEMISTS SOCIETY 84 (8) 749 - 754 0003-021X 2007/08 [Refereed][Not invited]
   

  The presence of conjugated trienoic acids were found in the purified fish oil from the Japanese sardine by an analysis of high performance liquid chromatography equipped with a photodiode-array detector. Gas chromatography-mass spectrometry indicated that main conjugated trienoic acids were those of 16:3, 20:5, and 22:6. Conjugated trienoic acid of 16:3 was identified as a cyclopropanoid acid with conjugated triene by NMR analysis. All conjugated trienoic acids were formed by heat treatment during the deodorization of fish oil, but not found after de-gumming, alkaline refining, and bleaching. Conjugated trienoic acids of 16:3, 20:5, and 22:6 originated from 16:4n-1, 20:5n-3 (EPA), and 22:6n-3 (DHA) in the fish oil during the heat treatment for deodorization.
• Oxidative stability of salmon and herring roe lipids and their dietary effect on plasma cholesterol levels of rats

  Hiromi Moriya, Takashi Kuniminato, Masahi Hosokawa, Kenji Fukunaga, Toshimasa Nishiyama, Kazuo Miyashita

  FISHERIES SCIENCE 73 (3) 668 - 674 0919-9268 2007/06 [Refereed][Not invited]
   

  The oxidative stability of lipids from salmon roe and herring roe was compared with those of commercial fish oils originated from sardine and tuna. Both fish roe lipids contained high amounts of eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA). Total EPA and DHA was more than 35% of total fatty acids in both roe lipids. On the basis of oxygen consumption, fish roe lipids showed the higher oxidative stability than both fish oils. This tendency in oxidative stabilities was also confirmed by the determination of propanal formation during oxidation. Analyses of lipid compositions suggested that the higher oxidative stabilities of fish roe lipids would be mainly due to the presence of phospholipids in them. Dietary effects of salmon and herring roe lipids were also determined. Little increase in total cholesterol level was observed in plasma lipids in rats fed salmon and herring roe lipids, although cholesterol content in fish roe lipids were 6.3% and 9.7% of total lipids for salmon roe and herring roe.
• Combination effect of herring roe lipids and proteins on plasma lipids and abdominal fat weight of mouse

  H. Moriya, M. Hosokawa, K. Miyashita

  JOURNAL OF FOOD SCIENCE 72 (5) C231 - C234 0022-1147 2007/06 [Refereed][Not invited]
   

  Dietary effects of herring roe lipids (HR-L) and proteins (HR-P) on plasma lipids and abdominal fat pad weight were determined. The main lipid class of HR-L was phospholipids (74%) and the main fatty acids were palmitic acid (16:0, 25.8%), DHA (22:6n-3, 21.6%), EPA (20:5n-3, 14.4%), and oleic acid (18:1n-9, 13.2%). A little increase in total cholesterol level was observed in plasma lipids of mouse fed with HR-L, although HR-L contained 9% cholesterol. This would be due to the lowering effect of EPA and DHA contained in HR-L on plasma cholesterol. Replacement of a part of dietary protein (5%) to HR-P reduced abdominal fat pad weight, but not significantly. On the other hand, combination of HR-P and HR-L significantly reduced the fat pad weight of the mouse as compared with the control. A sigificant effect of HR-P + HR-L was also observed in the reducing plasma lipid levels.
• Fucoxanthin and fucoxanthinol enhance the amount of docosahexaenoic acid in the liver of KKAy obese/diabetic mice

  Takayuki Tsukui, Kensuke Konno, Masashi Hosokawa, Hayato Maeda, Tokutake Sashima, Kazuo Miyashita

  JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY 55 (13) 5025 - 5029 0021-8561 2007/06 [Refereed][Not invited]
   

  This study examined the effect of dietary fucoxanthin or fucoxanthinol on the amount of docosahexaenoic acid (DHA) in the liver of KKAy mice, a model for obese/type II diabetes. In the first experiment, mice were fed diets containing crude fucoxanthin or glyceroglycolipid for 4 weeks. Results showed a significant increase in the level of DHA in mice fed 0.53% crude fucoxanthin, from 2.3% in control mice to 5.1% of fatty acid composition of total liver lipids. On the other hand, in mice fed crude glyceroglycolipid, the level of DHA as a proportion of total liver fatty acids remained unchanged. To clarify the enhancement of hepatic DHA, in the second experiment, KKAy mice were fed a diet containing purified fucoxanthin or its deacetylated derivative, fucoxanthinol. Results from a quantitative analysis using an internal standard showed that in mice fed 0.2% fucoxanthin, the amount of hepatic DHA was 2-fold higher than in control mice, whereas DHA levels in the small intestine remained unchanged. Furthermore, 0.2% fucoxanthinol led to 1.8- and 1.2-fold increases in the amount of hepatic DHA and arachidonic acid compared to control mice, respectively. These results indicate for the first time that dietary fucoxanthin and fucoxanthinol enhance the amount of DHA in the liver of KKAy mice.
• Therapy of myeloma in vivo using marine phospholipid in combination with agaricus blazei murill as immune respond activator.

  Kentaro Murakawa, Kenji Fukunaga, Masatoshi Tanouchi, Masashi Hosokawa, Zakir Hossain, Koretaro Takahashi

  J. Oleo Sci. Japan oil chemists' society 56 (4) 179 - 188 1345-8957 2007 [Refereed][Not invited]
   

  Mushroom (Agaricus blazei Murill) extract has been reported to possess antitumor effects through immune activation. Here, we investigated the beneficial effects of combining A. blazei extract with marine phospholipids in comparison to A. blazei extract alone on myeloma sp2 tumor suppression when orally administrated. The experimental groups designed for sp2 tumor bearing BALB/c nu/nu mice were drinks of: (1)control; (2)1.0 mg/mL squid phospholipid liposome alone; (3)0.5 mg/mL A. blazei Murill water extract alone; (4)1.0 mg/mL squid phospholipid liposome with 0.5 mg/mL A. blazei Murill water extract in the form of those simple mixture; and (5)1.0 mg/mL squid phospholipid liposome with 0.5 mg/mL A. blazei Murill water extract partially encapsulated. Orally administrated volumes amounted to approximately 5 mL per day per mouse for all groups. A. blazei Murill water extract alone and squid phospholipid alone served groups show moderate tumor suppression with total administrations of approximately 105 mg/mouse for squid phospholipid through out the experimental term. When both A. blazei Murill water extract and squid phospholipid were administrated simultaneously in a simple mixture form, promotional effect on cancer tumor suppression was observed. And when A. blazei Murill water extract was partially encapsulated in the squid phospholipid liposomes with total administrations being 105 mg/mouse for squid phospholipid, effect on cancer tumor suppression was more pronounced. Though there was no statistically significant difference in tumor sizes between the simple mixture form administrated group i.e. group (4) and the partially encapsulated form administrated group i.e. group (5), the tumor vanished mouse was seen in the partially encapsulated form administrated group. Thus it was concluded that combinational administration of the A. blazei Murill water extract and the marine phospholipid may be useful in myeloma sp2 therapy.
• Unsaturated phosphatidylethanolamine as effective synergist in combination with alpha-tocopherol.

  Takenaka Atsushi, Hosokawa Masashi, Miyashita Kazuo

  J. Oleo Sci. Japan Oil Chemists' Society 56 (10) 511 - 516 1345-8957 2007 [Refereed][Not invited]
   

  The effects of phosphatidylcholine (PC) and phosphatidylethanolamine (PE) on the oxidation of bonito oil triacylglycerol (TAG) were examined under the absence or presence of α-tocopherol. Synthesized PC or PE having saturated and unsaturated fatty acids in known position were used in this study. Unsaturated PC and PE promoted TAG oxidation under the absence of α-tocopherol. On the contrary, unsaturated PE showed synergistic antioxidant effect with α-tocopherol, while PC had little effect. The strongest synergistic effect was found by 1-palmitoyl (16:0, PA)-2-docosahexaenoyl (22:6n-3)-PE, followed by 1-PA-2-arachidonyl (20:4n-6)-PE, 1,2-dioleoyl (18:1n-9)-PE, and 1-PA-2-linoleoyl (18:2n-6)-PE, respectively.
  
• Effect of medium-chain triacylglycerols on anti-obesity effect of fucoxanthin.

  Maeda H, Hosokawa M, Sashima T, Funayama K, Miyashita K

  Journal of oleo science 12 56 (12) 615 - 621 1345-8957 2007 [Refereed][Not invited]
   

  Dietary effects of medium-chain triacylglycerols (MCT) and fucoxanthin (Fc) on abdominal fat weight were determined using KK-Ay obese mouse. Experimental diet contained MCT(0.9%), Fc (0.1%), or MCT (0.9%) +Fc (0.1%). The abdominal fat weight of mice fed with Fc was significantly lower than that of mice fed with MCT. Uncoupling protein 1 (UCP1), a key molecule for metabolic thermogenesis, was clearly expressed in the white adipose tissue (WAT) of mice fed Fc, but little expression in that of the mice fed MCT. The anti-obesity effect of Fc was increased by mixing Fc with MCT. This increase would be due to the increase in the absorption rate of Fc by MCT.
  
• Occurrence of conjugated linolenic acids in purified soybean oil

  Tomohisa Kinami, Naoto Horii, Bhaskar Narayan, Shingo Arato, Masashi Hosokawa, Kazuo Miyashita, Hironori Negishi, Junichi Ikuina, Ryuji Noda, Seiichi Shirasawa

  JOURNAL OF THE AMERICAN OIL CHEMISTS SOCIETY 84 (1) 23 - 29 0003-021X 2007/01 [Refereed][Not invited]
   

  A high-performance liquid chromatographic (HPLC) method is described for the determination of conjugated linoleic acids (CLA) and conjugated linolenic acids (CLN). Methyl esters prepared from purified lipid fractions of soybean oil were analyzed using an HPLC system equipped with photodiode-array detector to detect peaks having maximum absorption around 233 and 275 nm. These peaks were concentrated by AgNO3-silicic acid column chromatography and reversed-phase HPLC. The structural analysis, of dimethyloxazoline (DMOX) derivatized methyl esters, using gas chromatography-mass spectrometry (GC-MS) showed the occurrence of 9,11- and 10,12-CLA and 8,10,13-, 8,10,12-, and 9,11,13-CLN. The comparison of these conjugated fatty acids with authentic isomers by HPLC revealed the presence of isomeric mixtures of CLA [cis (c),trans(t) or t,c and t,t] and CLN (c,t,t or t,t,c and t,t,t). Traces of 9,11- and 10,12-CLA (c,t or t,c) were found in crude oil. CLN isomers (8,10,12-18:3 and 9,11,13-18:3) were found to be forming during the bleaching phase of soybean oil processing. 8,10,13-CLN and 9,11- and 10,12-CLA (t,t) were only found in soybean oil after the deodorization step. CLN contents in commercial soybean oil varied from 387 to 1,316 mg/kg oil. A decreased level of bleaching earth and temperature resulted in a reduced CLN content. It is possible that CLN would be derived from the linoleate hydroperoxides formed during the processing and storage of soybean oil.
• Unsaturated phosphatidylethanolamine as effective synergist in combination with alpha-tocopherol.

  Takenaka A, Hosokawa M, Miyashita K

  Journal of oleo science 10 56 (10) 511 - 516 1345-8957 2007 [Refereed][Not invited]
   

  The effects of phosphatidylcholine (PC) and phosphatidylethanolamine (PE) on the oxidation of bonito oil triacylglycerol (TAG) were examined under the absence or presence of α-tocopherol. Synthesized PC or PE having saturated and unsaturated fatty acids in known position were used in this study. Unsaturated PC and PE promoted TAG oxidation under the absence of α-tocopherol. On the contrary, unsaturated PE showed synergistic antioxidant effect with α-tocopherol, while PC had little effect. The strongest synergistic effect was found by 1-palmitoyl (16:0, PA)-2-docosahexaenoyl (22:6n-3)-PE, followed by 1-PA-2-arachidonyl (20:4n-6)-PE, 1,2-dioleoyl (18:1n-9)-PE, and 1-PA-2-linoleoyl (18:2n-6)-PE, respectively.
  
• 9trans,11trans conjugated linoleic acid inhibits the development of azoxymethane-induced colonic aberrant crypt foci in rats

  Yumiko Yasui, Rikako Suzuki, Hiroyuki Kohno, Shingo Miyamoto, Fumiaki Beppu, Masashi Hosokawa, Kazuo Miyashita, Takuji Tanaka

  NUTRITION AND CANCER-AN INTERNATIONAL JOURNAL 59 (1) 82 - 91 0163-5581 2007 [Refereed][Not invited]
   

  We investigated the effects of 9trans,11trans (9t,11t)-conjugated linoleic acid (CLA) isomer on azoxymethane (AOM)-induced colonic aberrant crypt foci (ACF) in rats. Male F344 rats were given 2 weekly subcutaneous injections of AOM (20 mg/kg bw) to induce colonic ACF. They also were fed a diet containing either 0.01%, 0.1%, or 1% 9t,11t-CLA for 4 wk starting 1 wk before the first dosing of AOM. The group that received a diet supplemented with 90 11t-CLA had a significantly lower number of ACF/colon in comparison to the AOM alone group in a dose-dependent manner up to 0.1%. Furthermore, treatment with 9t, 11t-CLA induced apoptosis and suppressed cell proliferation activity in the non-lesional crypts. The downregulation of cyclooxygenase-2 and cyclin D1 and the activation of peroxisome proliferators activated receptor gamma were observed in the colonic mucosa of rats fed a diet supplemented with 9t, 11t-CLA. Our findings thus provide some novel insight into the chemopreventive effect of 9t,11t-CLA against preinitiation as well as postinitiation stages of colorectal carcinogenesis.
• Potent inhibitory effect of trans9, trans 11 isomer of conjugated linoleic acid on the growth of human colon cancer cells

  Fumiaki Beppu, Masashi Hosokawa, Leo Tanaka, Hiroyuki Kohno, Takuji Tanaka, Kazuo Miyashita

  JOURNAL OF NUTRITIONAL BIOCHEMISTRY 17 (12) 830 - 836 0955-2863 2006/12 [Refereed][Not invited]
   

  This study compared the growth inhibitory effects of pure conjugated linoleic acid (CLA) isomers [cis(c)9,c11-CLA, c9,trans(t)11-CLA, t9,t11-CLA, and t10,c12-CLA] on human colon cancer cell lines (Caco-2, HT-29 and DLD-1). When Caco-2 cells were incubated up to 72 h with 200 mu M, each isomer, even in the presence of 10% fetal bovine serum (FBS), cell proliferation was inhibited by all CLA isomers in a time-dependent manner. The strongest inhibitory effect was shown by t9,t11-CLA, followed by t10,c12-CLA, c9,c11-CLA and c9,t11-CLA, respectively. The strongest effect of t9,t11-CLA was also observed in other colon cancer cell lines (HT-29 and DLD-1). The order of the inhibitory effect of CLA isomer was confirmed in the presence of 1% FBS. CLA isomers supplemented in the culture medium were readily incorporated into the cellular lipids of Caco-2 and changed their fatty acid composition. The CLA contents in cellular lipids were 26.2 +/- 2.7% for t9,t11-CLA, 35.9 +/- 0.3% for c9,t11-CLA and 46.3 +/- 0.8% for t10,c12-CLA, respectively. DNA fragmentation was clearly recognized in Caco-2 cells treated with t9,t11-CLA. This apoptotic effect of t9,t11-CLA was dose- and time-dependent. DNA fragmentation was also induced by 9c,11t-CLA and t10,c12-CLA. However, fragmentation levels with both isomers were much lower than that with t9,t11-CLA. t9t11-CLA treatment of Caco-2 cells decreased Bcl-2 levels in association with apoptosis, whereas Bax levels remained unchanged. These results suggest that decreased expression of Bcl-2 by t9t11-CLA might increase the sensitivity of cells to lipid peroxidation and to programmed cell death, apoptosis. (c) 2006 Elsevier Inc. All rights reserved.
• Catalpa seed oil rich in 9t,11t,13c-conjugated linolenic acid suppresses the development of colonic aberrant crypt foci induced by azoxymethane in rats

  Rikako Suzuki, Yumiko Yasui, Hiroyuki Kohno, Shingo Miyamoto, Masashi Hosokawa, Kazuo Miyashita, Takuji Tanaka

  ONCOLOGY REPORTS 16 (5) 989 - 996 1021-335X 2006/11 [Refereed][Not invited]
   

  Catalpa (Catalpa ovata) seed oil (CPO) is a unique oil that contains a high amount of 9trans,11trans,13cis-conjugated linolenic acid. In the present study, we investigated whether dietary administration with CPO affects the development of azoxymethane (AOM)-induced colonic aberrant crypt foci (ACF) in male F344 rats to elucidate its possible cancer chemopreventive efficiency. Also, the effect of CPO on the fatty acid composition of liver tissue and colonic mucosa, the serum levels of total cholesterol and triglyceride, and the mRNA expression of cyclooxygenase (COX)-2 in the colonic mucosa were measured. In addition, the cell proliferation activity and apoptotic index in the colonic mucosa were estimated immunohistochemically. Animals were given two weekly subcutaneous injections of AOM (20 mg/kg body weight). They also received the experimental diet containing 0.01 %, 0.1 % or 1 % CPO for 4 weeks, starting one week before the first dosing of AOM. AOM exposure produced a substantial number of ACF (99 28) at the end of the study (week 4). Dietary administration of CPO reduced the number of ACF (AOM + 0.01 % CPO, 32 +/- 11, P < 0.001; AOM + 0.1 % CPO, 35 +/- 18, P < 0.001; AOM + 1% CPO, 18 10, P < 0.001). 9t,11t-conjugated linoleic acid was detected in the liver tissue and colonic mucosa of rats fed the CPO-containing diet. Additionally, dietary administration with CPO decreased the serum triglyceride level and the expression of COX-2 mRNA in the colonic mucosa. The indices of cell proliferation and apoptosis in the colonic mucosa of rats treated with AOM and 1% CPO have significant differences when compared with the AOM alone group. These findings suggest the possible chemopreventive activity of CPO in the early phase of colon carcinogenesis.
• Production of phosphatidylcholine containing conjugated linoleic acid mediated by phospholipase A(2)

  Yukihiro Yamamoto, Masashi Hosokawa, Kazuo Miyashita

  JOURNAL OF MOLECULAR CATALYSIS B-ENZYMATIC 41 (3-4) 92 - 96 1381-1177 2006/08 [Refereed][Not invited]
   

  Esterification of lysophosphatidylcholine (LPC) with conjugated linoleic acid (CLA) was carried out using porcine pancreatic phospholipase A(2) (PLA(2)). PLA(2) only slightly synthesized phosphatidylcholine containing CLA (CLA-PC) at 2.6% by the addition of water. Addition of formamide in place of water markedly increased the yield of CLA-PC. In addition, synthesis of CLA-PC by PLA(2) was affected by the amount of substrate CLA and PLA(2) in the reaction system. Under optimal reaction conditions using 11 mg LPC, 18 mg CLA, 550 mg glycerol, 50 mu L formamide, 3.3 x 10(4) U PLA(2), and 0.3 mu mol CaCl2 at 37 degrees C for 6 h, the reaction yield of CLA-PC reached 65 mol%. Furthermore, addition of protein such as albumin and casein suppressed the decrease of CLA-PC yield after 6 h. PLA(2) exhibited the highest activity for the 10t,12c-CLA isomer among four CLA isomers (9c,11t-CLA,9c,11c-CLA,9t,11t-CLA and 10t,12c-CLA), whereas that for 9c,11c-CLA was the lowest. These results showed that the present esterification system for LPC and CLA by PLA(2) is effective for producing CLA-PC. (c) 2006 Elsevier B.V. All rights reserved.
• 脂肪酸代謝に対する海藻脂質の影響

  津久井 隆行, 今野 賢亮, 細川 雅史, 宮下 和夫

  脂質栄養学 日本脂質栄養学会 15 (2) 175 - 175 1343-4594 2006/08
• Production of docosahexaenoic acid bounded phospholipids via phospholipase A(2) mediated bioconversion

  Shinichi Awano, Koji Miyamoto, Masashi Hosokawa, Mitsumasa Mankura, Koretaro Takahashi

  FISHERIES SCIENCE 72 (4) 909 - 911 0919-9268 2006/08 [Refereed][Not invited]
  
• Fucoxanthin and its metabolite, fucoxanthinol, suppress adipocyte differentiation in 3T3-L1 cells

  Hayato Maeda, Masashi Hosokawa, Tokutake Sashima, Nobuyuki Takahashi, Teruo Kawada, Kazuo Miyashita

  INTERNATIONAL JOURNAL OF MOLECULAR MEDICINE 18 (1) 147 - 152 1107-3756 2006/07 [Refereed][Not invited]
   

  Fucoxanthin is a major carotenoid found in edible seaweed such as Undaria pinnatifida and Hijikia fusiformis. We investigated the suppressive effects of fucoxanthin and its metabolite, fucoxanthinol, on the differentiation of 3T3-L1 preadipocytes to adipocytes. Fucoxanthin inhibited intercellular lipid accumulation during adipocyte differentiation of 3T3-L1 cells. Furthermore, fucoxanthin was converted to fucoxanthinol in 3T3-L1 cells. Fucoxanthinol also exhibited suppressive effects on lipid accumulation and decreased glycerol-3-phosphate dehydrogenase activity, an indicator of adipocyte differentiation. The suppressive effect of fucoxanthinol was stronger than that of fucoxanthin. In addition, in 3T3-L1 cells treated with fucoxanthin and fucoxanthinol, peroxisome proliferator-activated receptor gamma (PPAR gamma), which regulates adipogenic gene expression, was down-regulated in a dose-dependent manner. These results suggest that fucoxanthin and fucoxanthinol inhibit the adipocyte differentiation of 3T3-L1 cells through down-regulation of PPAR gamma. Fucoxanthinol had stronger suppressive effects than fucoxanthin on adipocyte differentiation in 3T3-L1 cells.
• Growth inhibition and apoptosis induction by all-trans-conjugated linolenic acids on human colon cancer cells

  Yumiko Yasui, Masashi Hosokawa, Hiroyuki Kohno, Takuji Tanaka, Kazuo Miyashita

  ANTICANCER RESEARCH 26 (3A) 1855 - 1860 0250-7005 2006/05 [Refereed][Not invited]
   

  Conjugated linolenic acids (CLN) are geometric and positional isomers of linolenic acid. Growth inhibition and apoptosis induction by a-eleostearic acid (c.9,t11,t13-CLN), P-eleostearic acid (t9, t11, t13-CLN), a-calendic acid (t8.t10, c12-CLN) and P-calendic acid (t8, t10, t12-CLN) were compared. beta-Eleostearic acid and P-calendic acid, which have all-transconjugated double bonds, exerted stronger growth inhibition and more DNA fragmentation, an indicator of apoptosis induction, in the human colon cancer cells Caco-2 than a-eleostearic acid and a-calendic acid with the cis configuration. Down-regulation of bcl-2 and up-regulation of bax mRNA by beta-eleostearic acid were also greater than by a-eleostearic acid. Interestingly, the cytotoxic effects of beta-eleostearic acid and beta-calendic acid. were not counteracted completely by a-tocopherol. whereas the cytotoxic effects of a-eleostearic and a-calendic acids were lost in the presence of a-tocopherol. These results suggest that P-eleostearic and beta-calendic acids have signaling pathways different from those of a-eleostearic and a-calendic acids and exhibit high potency for reducing the cell viability of Caco-2.
• Growth inhibition and apoptosis induction by all-trans-conjugated linolenic acids on human colon cancer cells

  Yumiko Yasui, Masashi Hosokawa, Hiroyuki Kohno, Takuji Tanaka, Kazuo Miyashita

  Anticancer Research 26 (3 A) 1855 - 1860 0250-7005 2006/05 [Refereed][Not invited]
   

  Conjugated linolenic acids (CLN) are geometric and positional isomers of linolenic acid. Growth inhibition and apoptosis induction by α-eleostearic acid (c9,t11,t13-CLN), β-eleostearic acid (t9,t11,t13-CLN), α-calendic acid (t8,t10,c12-CLN) and β-calendic acid (t8,t10,t12-CLN) were compared, β-Eleostearic acid and β-calendic acid, which have all-trans-conjugated double bonds, exerted stronger growth inhibition and more DNA fragmentation, an indicator of apoptosis induction, in the human colon cancer cells Caco-2 than α-eleostearic acid and α-calendic acid with the cis configuration. Down-regulation of bcl-2 and up-regulation of bax mRNA by β-eleostearic acid were also greater than by α-eleostearic acid. Interestingly, the cytotoxic effects of β-eleostearic acid and β-calendic acid were not counteracted completely by α-tocopherol, whereas the cytotoxic effects of α-eleostearic and α-calendic acids were lost in the presence of α-tocopherol. These results suggest that β-eleostearic and β-calendic acids have signaling pathways different from those of α-eleostearic and α-calendic acids and exhibit high potency for reducing the cell viability of Caco-2.
• Inhibition properties of dipeptides from salmon muscle hydrolysate on angiotensin I-converting enzyme

  S Ono, M Hosokawa, K Miyashita, K Takahashi

  INTERNATIONAL JOURNAL OF FOOD SCIENCE AND TECHNOLOGY 41 (4) 383 - 386 0950-5423 2006/04 [Refereed][Not invited]
   

  We isolated Phe-Leu as an angiotensin I-converting enzyme (ACE) inhibitor from hydrolysate of chum salmon muscle. The IC50 value of this peptide was 13.6 mu M, and it showed non-competitive inhibition. The reverse sequence dipeptide Leu-Phe also showed ACE inhibitory activity. However, Leu-Phe is much less inhibitory than Phe-Leu with an IC50 value of 383.2 mu M. In addition, the inhibition mode was competitive. To investigate the relationship between dipeptide sequence and ACE inhibition properties, we further measured ACE inhibitory activity and inhibition mechanism using six Trp-containing dipeptides, which had been identified from the same salmon muscle hydrolysate as ACE inhibitory peptides in a previous study. Peptides with Trp as the C-terminal residue, Ala-Trp, Val-Trp, Met-Trp, Ile-Trp, Leu-Trp showed non-competitive inhibition. On the other hand, reversed sequence peptides with Trp at the N-terminal were competitive inhibitors, except Trp-Leu. These results indicate that the sequence of ACE inhibitory dipeptides can affect both inhibitory potency and the inhibition mechanisms.
• Docosahexaenoic acid and eicosapentaenoic acid-enriched phosphatidylcholine liposomes enhance the permeability, transportation and uptake of phospholipids in Caco-2 cells

  Zakir Hossain, Hideyuki Kurihara, Masashi Hosokawa, Koretaro Takahashi

  MOLECULAR AND CELLULAR BIOCHEMISTRY 285 (1-2) 155 - 163 0300-8177 2006/04 [Refereed][Not invited]
   

  The influence of docosahexaenoic acid (DHA)- and eicosapentaenoic acid (EPA)-enriched phosphatidylcholine (PC) on the permeability, transport and uptake of phospholipids was evaluated in Caco-2 cells. The cells were grown on permeable polycarbonate transwell filters, thus allowing separate access to the apical and basolateral chambers. The monolayers of the cells were used to measure lucifer yellow permeability and transepithelial electrical resistance (TEER). Transcellular transportation of diphenylhexatriene (DPH) labeled-PC small unilamellar vesicles (SUV) from the apical to basolateral chamber, and uptake of the same SUV was monitored in the cell monolayers. Cell-membrane perturbation was evaluated to measure the release of lactate dehydrogenase and to determine the cell viability with sodium 2-(4-iodophenyl)-3-(4-nitrophenyl) -5-(2, 4-disulfophenyl)-2H-tetrazolium dye reduction assay. The lucifer yellow flux was 1.0 and 1.5 nmol/h/cm(2)with 50 mu M PC, and 17.0 and 23.0 nmol/h/cm(2) with 100 mu M PC when monolayers of Caco-2 cells were treated with DHA- and EPA-enriched PC, respectively. TEER decreased to 24 and 27% with 50 and 100 mu M DHA-enriched PC, and to 25 and 30% with 50 and 100 mu M EPA-enriched PC, respectively. Our results show that DHA- and EPA-enriched PC increases tight junction permeability across the Caco-2 cell monolayer whereas soy PC has no effect on tight junction permeability. Transportation and uptake of DHA- and EPA-enriched PC SUV differed significantly (P < 0.01) from those of soy PC SUV at all doses. We found that PC SUV transported across Caco-2 monolayer and was taken up by Caco-2 cells with very slight injury of the cell membrane up to 100 mu M PC. Lactate dehydrogenase release and cell viability did not differ significantly between the treatment and control, emphasizing that injury was minimal. Our results suggest that DHA- and EPA-enriched PC enhance the permeability, transport and uptake of PC SUV across monolayers of Caco-2 cells.
• Effect of polyunsaturated fatty acid-enriched phosphatidylcholine and phosphatidylserine on butyrate-induced growth inhibition, differentiation and apoptosis in Caco-2 cells

  Z Hossain, M Konishi, M Hosokawa, K Takahashi

  CELL BIOCHEMISTRY AND FUNCTION 24 (2) 159 - 165 0263-6484 2006/03 [Refereed][Not invited]
   

  Phospholipids are fascinating in terms of important bio-functional compounds. The present work investigated the effect of polyunsaturated phosphatidylcholine (PC) and phosphatidylserine (PS) on butyrate-induced growth inhibition, differentiation and apoptosis using Caco-2 cells. Growth inhibition of Caco-2 cells became apparent 24 h after addition of PC while it took 48 h with PS. Alkaline phosphatase activity of Caco-2 cells increased with combined PC or PS and sodium butyrate (NaBT) at 72 h, indicating that PC and PS enhanced cell differentiation in the presence of NaBT. An increased enrichment factor was also found when cells were treated with combinations of PC or PS and NaBT. These results suggest that marine PC and PS can be considered to be potentially useful colon cancer chemotherapy agents with high bio-availability. Copyright (c) 2005 John Wiley & Sons, Ltd.
• Production of value added phospholipids via phospholipase A2 mediated bioconversion from soy lysolecithin and docosahexaenoic acid rich free fatty acid mixture.

  Fisheries Sci., 72 909 - 912 2006 [Refereed][Not invited]
  
• Troglitazone and 9cis, 11trans, 13trans-conjugated linolenic acid: Comparison of their antiproliferative and apoptosis-inducing effects on different colon cancer cell lines

  Y. Yasui, M. Hosokawa, H. Kohno, T. Tanaka, K. Miyashita

  CHEMOTHERAPY 52 (5) 220 - 225 0009-3157 2006 [Refereed][Not invited]
   

  Background. We have previously reported that troglitazone, a synthetic ligand for peroxisome proliferator-activated receptor gamma (PPAR gamma), and bitter gourd seed oil rich in 9cis,11trans,13trans-conjugated linolenic acid (9c,11t,13t-CLN) prevent colon carcinogenesis. To evaluate the chemotherapeutic effect and potency of these compounds on colon cancer cells, we investigated their antiproliferative and apoptosis-inducing effects using different human colon cancer cell lines. Methods: The antiproliferative and apoptosis-inducing effects of troglitazone and 9c,11t,13t-CLN were evaluated and compared using HT-29, DILD-11 and Caco-2 cells at different stages of enterocytic differentiation. Results: Troglitazone and 9c,11t,13t-CLN decreased cell viability and induced apoptosis in three colon cancer cell lines. The susceptibility of HT-29, which expresses IPIPAR gamma at high levels, to troglitazone and 9c,11t,13t-CLN was higher than that of Caco-2 cells with low levels of PPAR gamma. Con- clusion:Troglitazone and 9c,11 1 t,13t-CLN exhibited more effective chemotherapeutic effects on HT-29 cells than on Caco-2 cells. Copyright (c) 2006 S. Karger AG, Basel.
• Halocynthiaxanthin and fucoxanthinol isolated from Halocynthia roretzi induce apoptosis in human leukemia, breast and colon cancer cells

  Izumi Konishi, Masashi Hosokawa, Tokutake Sashima, Hidetaka Kobayashi, Kazuo Miyashita

  Comparative Biochemistry and Physiology Part C: Toxicology & Pharmacology 142 (1-2) 53 - 59 1532-0456 2006/01 [Refereed][Not invited]
   

  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 mu M halocynthiaxanthin and fucoxanthinol was decreased by 12.1% and 5.7% of control after 48 h incubation, respectively. Furthermore, halccynthiaxanthin 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 mu M 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. Facoxanthinol 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. (c) 2005 Elsevier Inc. All right; reserved.
• Bitter gourd seed fatty acid rich in 9c,11t,13t-conjugated linolenic acid induces apoptosis and up-regulates the GADD45, p53 and PPAR gamma in human colon cancer Caco-2 cells

  Y Yasui, M Hosokawa, T Sahara, R Suzuki, S Ohgiya, H Kohno, T Tanaka, K Miyashita

  PROSTAGLANDINS LEUKOTRIENES AND ESSENTIAL FATTY ACIDS 73 (2) 113 - 119 0952-3278 2005/08 [Refereed][Not invited]
   

  Bitter gourd (Momordica charantia) seed oil (BGO) is a unique oil which contains 9cis, 11trans, 13trans-conjugated linolenic acid (9c,11t,13t-CLN) at a high level of more than 60%. In this study, we investigated the anti-proliferative and apoptosis-inducing effects of free fatty acids prepared from BGO (BGO-FFA) using colon cancer Caco-2 cells. BGO-FFA and purified 9c,11t,13t-CLN remarkably reduced the cell viability of Caco-2. In Caco-2 cells treated with BGO-FFA, DNA fragmentation of apoptosis indicators was observed in a dose-dependent manner. The expression level of apoptosis suppressor Bcl-2 protein was also decreased by BGO-FFA treatment. The GADD45 and p53, which play an important role in apoptosis-inducing pathways, were remarkably up-regulated by BGO-FFA treatment in Caco-2 cells. Up-regulation of PPAR gamma mRNA and protein were also observed during apoptosis induced by BGO-FFA. These results suggest that BGO-FFA rich in 9c,11t,13t-CLN may induce apoptosis in Caco-2 cells through up-regulation of GADD45, p53 and PPAR gamma. (C) 2005 Elsevier Ltd. All rights reserved.
• Fucoxanthin from edible seaweed, Undaria pinnatifida, shows antiobesity effect through UCP1 expression in white adipose tissues

  H Maeda, M Hosokawa, T Sashima, K Funayama, K Miyashita

  BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS 332 (2) 392 - 397 0006-291X 2005/07 [Refereed][Not invited]
   

  Mitochondrial uncoupling protein 1 (UCP1) is usually expressed only in brown adipose tissue (BAT) and a key molecule for metabolic thermogenesis to avoid an excess of fat accumulation. However, there is little BAT in adult humans. Therefore, UCP1 expression in tissues other than BAT is expected to reduce abdominal fat. Here, we show reduction of abdominal white adipose tissue (WAT) weights in rats and mice by feeding lipids from edible seaweed, Undaria pinnatifida. Clear signals of UCP1 protein and mRNA were detected in WAT of mice fed the Undaria lipids, although there is little expression of UCP1 in WAT of mice fed control diet. The Undaria lipids mainly consisted of glycolipids and seaweed carotenoid, fucoxanthin. In the fucoxanthin-fed mice, WAT weight significantly decreased and UCP1 was clearly expressed in the WAT, while there was no difference in WAT weight and little expression of UCP1 in the glycolipids-fed mice. This result indicates that fucoxanthin upregulates the expression of UCP1 in WAT, which may contribute to reducing WAT weight. (C) 2005 Elsevier Inc. All rights reserved.
• Docosahexaenoic acid-containing phosphatidylethanolamine enhances HL-60 cell differentiation by regulation of c-jun and c-myc expression

  H Ishigamori, M Hosokawa, H Kohno, T Tanaka, K Miyashita, K Takahashi

  MOLECULAR AND CELLULAR BIOCHEMISTRY 275 (1-2) 127 - 133 0300-8177 2005/07 [Refereed][Not invited]
   

  18:1/docosahexaenoic acid (DHA)-containing phosphatidylethanolamine (PE) enhanced cell differentiation and growth inhibition of HL-60 induced by dibutyryl cAMP (dbcAMP) in a dose-dependent manner. The combined treatment of 200 mu M dbcAMP and 50 mu M 18:1/DHA-PE increased the NBT reducing activity, which is as an indicator of cell differentiation, to more than 75% from 40% of cells treated with 200 mu M dbcAMP alone. In HL-60 cells treated with 50 mu M 18:1/DHA-PE and 200 mu M dbcAMP for 24 h, the expression level of c-jun mRNA and c-Jun protein were remarkably elevated compared to cells treated with dbcAMP alone. In contrast, there was no difference in the expression levels of c-fos mRNA and c-Fos protein between the combination of 18:1/DHA-PE + dbcAMP or dbcAMP alone. On the other hand, the combine treatment of 18:1/DHA-PE and dbcAMP markedly reduced the expression level of c-myc oncogene during 48 h incubation. The decreases of c-myc mRNA by 18:1/DHA-PE and/or dbcAMP was correlated with growth inhibition effect. Thus, 18:1/DHA-PE might enhance dbcAMP-induced HL-60 cell differentiation and growth inhibition by regulation of c-jun and c-myc mRNA and their products.
• Lipid peroxidation of a human hepatoma cell line (HepG2) after incorporation of linoleic acid, arachidonic acid, and docosahexaenoic acid

  M Araseki, H Kobayashi, M Hosokawa, K Miyashita

  BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY 69 (3) 483 - 490 0916-8451 2005/03 [Refereed][Not invited]
   

  Lipid peroxidation of human heptoma cell line, HepG2, after incorporation of linoleic acid (LA), arachidonic acid (AA), and docosahexaenoic acid (DHA) was measured with a fluorescent probe and gas chromatography-mass spectrometry (GC-MS) analysis. The analysis with a fluorescent probe showed that incorporation of each polyunsaturated fatty acid (PUFA) enhanced the cellular lipid peroxidation level, but there was little difference in the effect of LA, AA, or DHA on the enhancement of cellular lipid peroxidation. The fluorescent analysis also showed that the addition of H2O2 (0.5 mm) enhanced the cellular lipid peroxidation levels in LA and AA supplemented cells as compared with those without H2O2. However, the enhancement of lipid peroxidation by H2O2 was not observed in DHA-supplemented cells. The same result was obtained in the GC-MS analysis of total amounts of monohydroperoxides (MHP) formed in the cellular phospholipid oxidation. In this case, the main source for MHP was LA in LA-, AA-, and DHA-supplemented cells. A significant amount of AA-MHP and a small amount of DHA-MHP were observed in AA- and DHA-supplemented cells respectively. GC-MS analysis also indicated the specific positional distribution of DHA-MHP isomers. The isomers were formed only by hydrogen abstraction at the C-18 (16-MHP + 20-MHP; 46.5%), C-6 (4-MHP + 8-MHP; 38.5%), and C-12 (10-MHP + 14-MHP; 15.1%) positions, but not at the C-9 or C-15 positions.
• Butyrate-Induced Growth Inhibition and Differentiation in Caco-2 Cells with Hybrid Liposomes

  Zakir Hossain, Hideyuki Kurihara, Masashi Hosokawa, Koretaro Takahashi

  Journal of Oleo Science 54 (5) 281 - 288 1347-3352 2005 [Refereed][Not invited]
   

  Hybrid liposomes were prepared by mixing glycerophospholipids with glyceroglycolipids. The effects of hybrid liposomes on Caco-2 cells were investigated. Growth inhibition of cells was determined with sodium 2-(4-iodophenyl)-3-(4-nitrophenyl)-5-(2, 4-disulfophenyl)-2H-tetrazolium dye reduction assay. Alkaline phosphatase activity was measured for assessing the cell differentiation induced by sodium butyrate (NaBT). Growth of Caco-2 cells was inhibited at 48 h after addition of all four types hybrid liposomes (PC/DGDG, PC/SQDG, PS/DGDG and PS/SQDG) while at 72 h growth inhibition was significant for all treatments except the hybrid liposome of phosphatidylcholine and digalactosyldiacylglycerol under concentrations of 50 μM glycerophospholipids with 50 μM glyceroglycolipids. Alkaline phosphatase activity of Caco-2 cells increased significantly when cells were treated in combination of hybrid liposomes and NaBT at 72 h, indicating that hybrid liposomes enhanced cell differentiation induced with NaBT. © 2005, Japan Oil Chemists' Society. All rights reserved.
• Araseki, M., Kobayashi, H., Hosokawa, M. and Miyashita. K.:"Lipid peroxidation of a human hepatoma cell line (HepG2) after incorporated linoleic acid, arachidonic acid, and docosahexaenoic acid", Biosci. Biotechnol. Biochem., 69:483-490(2005)*

  2005 [Refereed][Not invited]
  
• Ishigamori, H., Hosokawa, M., Kohno, H., Tanaka, T., Miyashita, K. and Takahashi, K.:"DHA-containing phosphatidylethanolamine enhances HL-60 cell differentiation through alterations of c-jun and c-myc expression", Mol. Cell. Biochem., 275:127-133(2005)*

  2005 [Refereed][Not invited]
  
• Growth inhibition and induction of differentiation and apoptosis mediated by sodium butyrate in Caco-2 cells with algal glycolipids

  Z Hossain, H Kurihara, M Hosokawa, K Takahashi

  IN VITRO CELLULAR & DEVELOPMENTAL BIOLOGY-ANIMAL 41 (5-6) 154 - 159 1071-2690 2005 [Refereed][Not invited]
   

  Glycolipids should have potential effects as antitumor agents. However. very few studies have examined this property of digalactosyl diacylglycerol (DGDG) and sulfoquinovosyl diacylglycerol (SQDG) on colon cancer cells. Cell viability was determined every 24 h with sodium 2-(4-iodophenyl)-3-(4 -nitrophenyl)-5-(2, 4-disulfophenyl)-2H-tetrazolium dye reduction assay up to 72 h. Alkaline phosphalase activity was measured for assessing cell differentiation. Apoptosis was tested with enzyme-linked immunosorbent assay analysis. Growth of Caco-2 cells was inhibited apparently at 48 h after addition of SQDG and at 72 h with DGDG. Alkaline phosphalase activity of Caco-2 cells obviously increased in combination with DGDG or SQDG and sodium butyrate (NaBT) at 72 h, indicating that DGDG and SQDG enhanced cell differentiation induced with NaBT. An increased enrichment factor was found when the cell was treated in combination with DGDG or SQDG and NaBT. These results Strongly suggest that DGDG and SQDG should he considered as the leading compounds of potentially useful colon cancer Chemotherapy agents when NaBT is combined.
• Growth inhibition of human pro-myelocytic leukemia (HL-60) cells by lipid extracts of marine alga Sargassum marginatum (Fucales, Phaeophyta) harvested off Goa (west coast of India) with special reference to fatty acid composition

  N Bhaskar, M Hosokawa, K Miyashita

  INDIAN JOURNAL OF MARINE SCIENCES 33 (4) 355 - 360 0379-5136 2004/12 [Refereed][Not invited]
   

  The present investigation deals with the fatty acid composition of total lipids and different lipid classes of brown seaweed Sargassum marginatum harvested off Goa. An effort was made to screen the growth inhibitiory/cytotoxic activity of lipid extracts on the human pro-melocytic leukemia (HL-60) cells. Phospholipids (PL) were found to be the most effective compared to the other lipid classes in terms of cytotoxic activity. PL exhibited cytotoxic activity at concentrations as low as 20 mug/ml. Phospholipids were found to be higher in poly unsaturated fatty acids (PUFA) among all the lipid classes analysed. This study indicates the possibility of seaweeds as potential sources of anticancer substances. Further works are needed to identify the active compound responsible for this anti-cancerous activity. The role of non-methylene interrupted (NMI) fatty acid specific to Sargassum spp, as anti-cancerous substance, also needs to be elucidated.
• Fucoxanthin induces apoptosis and enhances the antiproliferative effect of the PPARgamma ligand, troglitazone, on colon cancer cells.

  Hosokawa M, Kudo M, Maeda H, Kohno H, Tanaka T, Miyashita K

  Biochimica et biophysica acta 1-3 1675 (1/3) 113 - 119 0005-2728 2004/11 [Refereed][Not invited]
  
• Fucoxanthin induces apoptosis and enhances the antiproliferative effect of the PPARgamma ligand, troglitazone, on colon cancer cells.

  Hosokawa M, Kudo M, Maeda H, Kohno H, Tanaka T, Miyashita K

  Biochimica et biophysica acta 1675 (1-3) 113 - 119 0006-3002 2004/11 [Refereed][Not invited]
  
• Dietary seed oil rich in conjugated linolenic acid from bitter melon inhibits azoxymethane-induced rat colon carcinogenesis through elevation of colonic PPAR gamma expression and alteration of lipid composition

  H Kohno, Y Yasui, R Suzuki, M Hosokawa, K Miyashita, T Tanaka

  INTERNATIONAL JOURNAL OF CANCER 110 (6) 896 - 901 0020-7136 2004/07 [Refereed][Not invited]
   

  Our previous short-term experiment demonstrated that seed oil from bitter melon (Momordica charantia) (BMO), which is rich in cis(c)9, trans(t)11, t13-conjugated linolenic acid (CLN), inhibited the development of azoxymethane (AOM)-induced colonic aberrant crypt foci (ACF). In our study, the possible inhibitory effect of dietary administration of BMO on the development of colonic neoplasms was investigated using an animal colon carcinogenesis model initiated with a colon carcinogen AOM. Male F344 rats were given subcutaneous injections of AOM (20 mg/kg body weight) once a week for 2 weeks to induce colon neoplasms. They also received diets containing 0.01%, 0.1% or 1% BMO for 32 weeks, starting 1 week before the first dosing of AOM. At the termination of the study (32 weeks), AOM induced 83% incidence (15/18 rats) of colonic adenocarcinoma. Dietary supplementation with 0.01% and 0.1% BMO caused significant reduction in the incidence (47% inhibition by 0.01% BMO, p<0.02; 40% inhibition by 0.1% BMO, p<0.05; and 17% inhibition by 1% BMO) and the multiplicity (64% inhibition by 0.01% BMO, p<0.005; 58% inhibition by 0.1% BMO, p<0.02; and 48% inhibition by 1% BMO, p<0.05) of colonic adenocarcinoma, though a clear dose response was not observed. Such inhibition was associated with the increased content of CLA (c9,t11-18:2)in the lipid composition in colonic mucosa and liver. Also, BMO administration in diet enhanced expression of peroxisome proliferator-activated receptor (PPAR) gamma protein in the non-lesional colonic mucosa. These findings suggest that BMO rich in CLN can suppress AOM-induced colon carcinogenesis and the inhibition might be caused, in part, by modification of lipid composition in the colon and liver and/or increased expression of PPARgamma protein level in the colon mucosa. (C) 2004 Wiley-Liss, Inc.
• Pomegranate seed oil rich in conjugated linolenic acid suppresses chemically induced colon carcinogenesis in rats

  H Kohno, R Suzuki, Y Yasui, M Hosokawa, K Miyashita, T Tanaka

  CANCER SCIENCE 95 (6) 481 - 486 1347-9032 2004/06 [Refereed][Not invited]
   

  Pomegranate (Punica granatum L.) seed oil (PGO) contains more than 70% cis(c),trans(t)11, c13-18:3 as conjugated linolenic acids (CLN). Our previous short-term experiment demonstrated that seed oil from bitter melon (Momordica charantia) (BMO), which is rich in c9, t11, t13-CLN, inhibited the occurrence of colonic aberrant crypt foci (ACF) induced by azoxymethane (AOM). In this study, we investigated the effect of dietary PGO on the development of AOM-induced colonic malignancies and compared it with that of conjugated linoleic acid (CLA). To induce colonic tumors, 6-week old male F344 rats were given subcutaneous injections of AOM (20 mg/kg body weight) once a week for 2 weeks. One week before the AOM treatment they were started on diet containing 0.01%, 0.1%, or 1% PGO or 1% CLA for 32 weeks. Upon termination of the bioassay (32 weeks) colon tumors were evaluated histopathologically. AOM exposure produced colonic adenocarcinoma with an incidence of 81% and multiplicity of 1.88 +/- 1.54 at week 32. Administration of PGO in the diet significantly inhibited the incidence (AOM + 0.01% PGO, 44%, P < 0.05; AOM + 0.1% PGO, 38%, P < 0.01; AOM + 1% PGO, 56%) and the multiplicity (AOM + 0.01% PGO, 0.56 +/- 0.73, P < 0.01; AOM + 0.1% PGO, 0.50 +/- 0.73, P < 0.005; AOM + 1% PGO, 0.88 +/- 0.96, P < 0.05) of colonic adenocarcinomas, although a clear dose-response relationship was not observed at these dose levels. CLA feeding also slightly, but not significantly, reduced the incidence and multiplicity of colonic adenocarcinomas. The inhibition of colonic tumors by PGO was associated with an increased content of CLA (c9, t11, 18:2) in the lipid fraction of colonic mucosa and liver. Also, administration of PGO in the diet elevated expression of peroxisome proliferator-activated receptor (PPAR) gamma protein in the non-tumor mucosa. These results suggest that PGO rich in c9, t11, c13-CLN can suppress AOM-induced colon carcinogenesis, and the inhibition is associated in part with the increased content of CLA in the colon and liver and/or increased expression of PPARgamma protein in the colon mucosa.
• Separation of sardine oil without heating from surimi waste and its effect on lipid metabolism in rats

  K Toyoshima, R Noguchi, M Hosokawa, K Fukunaga, T Nishiyama, R Takahashi, K Miyashita

  JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY 52 (8) 2372 - 2375 0021-8561 2004/04 [Refereed][Not invited]
   

  Sardine oil was obtained by centrifugation of surimi wastewater without heating or chemical refining. This oil (CE) showed light yellow color and the peroxide value was less than 1.0 meq/kg. The main lipid class of CE was triacylglycerol (TG) (> 99%). These features indicate that CE can be directly used as food materials without further purification. Commercial sardine oil (CO) is usually prepared via some kind of refining process with high temperature (250 C) and chemical treatment. The comparative study on the physiological effects of these sardine oils (CE and CO) revealed that the dietary sardine oils were more effective in reducing abdominal fat pads, plasma total cholesterol, and TG levels of rats than was a soybean oil diet (control). Furthermore, these effects were greater in CE than CO, although there was little difference in the fatty acid composition of both oils. Although the main lipid class of CE was TG (> 99%), CE was prepared by centrifugation from surimi waste and directly used as dietary fat without further purification. Therefore, CE may contain some kinds of minor components, which could be attributed to the higher physiological activity of CE. To reveal the involvement of the minor compounds in CE, we prepared TG from CE by column chromatography and measured its effect on lipid metabolism of rats. TG from CE also showed the reducing effects on abdominal fad pads and plasma lipid levels. The effect of TG from CE was almost the same as that of original CE, suggesting that the higher nutritional activity of CE than CO may not be due to the minor compounds in CE.
• Bioconversion of n-3 and n-6 PUFA by Clavibacter sp ALA2

  M Hosokawa, CT Hou, D Weisleder

  JOURNAL OF THE AMERICAN OIL CHEMISTS SOCIETY 80 (11) 1085 - 1091 0003-021X 2003/11 [Refereed][Not invited]
   

  Clavibacter sp. ALA2 oxidized n-3 and n-6 PUFA into a variety of oxylipins. Structures of products converted from EPA and DHA were determined as 15,18-dihydroxy-14,17-epoxy-5(Z),8(Z),11(Z)-eicosatrienoic acid and 17,20-dihydroxy-16,19-epoxy-4(Z),7(Z),10(Z),13(Z)-docosatetraenoic acid by GC-MS and NMR analyses. In contrast, gamma-linolenic acid and arachidonic acid were converted to diepoxy bicyclic FA, tetrahydrofuranyl monohydroxy FA, and trihydroxy FA. Thus, the structures of bioconversion products were different between n-3 and n-6 PUFA. Furthermore, strain ALA2 placed hydroxy groups and cyclic structures at the same position from the omega-terminal despite the number of carbons in the chain and the double bonds in the PUFA.
• Biosynthesis of tetrahydrofuranyl fatty acids from linoleic acid by Clavibacter sp ALA2

  M Hosokawa, CT Hou, D Weisleder, W Brown

  JOURNAL OF THE AMERICAN OIL CHEMISTS SOCIETY 80 (2) 145 - 149 0003-021X 2003/02 [Refereed][Not invited]
   

  Clavibacter sp, ALA2 converts linoleic acid into many novel oxygenated products including hydroxy FA and tetrahydrofuranyl unsaturated FA (THFA). One of them was tentatively identified by CC-MS as 12,13,16-trihydroxy-9(Z)-oc-tadecenoic acid (12,13,16-THOA) (Hou, C.T., H.W. Gardner, and W. Brown, J Am. Oil Chem. Soc. 78:1167-1169, 2001). We have separated and purified 12,13,1 6-THOA from its isomer, 12,13,17-THOA, by silica gel column chromatography and by preparative TLC. Its structure was then confirmed by proton and C-13 NMR analyses, Purified 12,13,1 6-THOA was used as a substrate to study the biosynthesis of THFA. Within 24 h of incubation, cells of strain ALA2 converted 12,13,1 6-THOA to both 12-hydroxy-13,16-epoxy-9(Z)-octadecenoic acid (12-hydroxy-THFA) and 7,12-dihydroxy-13,16-epoxy-9(Z)-octadecenoic acid (7,12-dihydroxy-THFA). The relative abundance of 7,12-dihydroxy-THFA increased with incubation time, whereas that of 12,13,1 6-THOA and of 12-hydroxy-THFA decreased. Therefore, the biosynthetic pathway of THFA from linoleic acid by strain ALA2 is as follows: linoleic acid --> 12,13-dihydroxy-9(Z)-octadecenoic acid --> 12,13,16-THOA --> 12-hydroxy-THFA --> 7,12-dihydroxy-THFA.
• Isolation of peptides with angiotensin I-converting enzyme inhibitory effect derived from hydrolysate of upstream chum salmon muscle

  S Ono, M Hosokawa, K Miyashita, K Takahashi

  JOURNAL OF FOOD SCIENCE 68 (5) 1611 - 1614 0022-1147 2003 [Refereed][Not invited]
   

  In order to utilize upstream chum salmon as a component of nutraceutical food, their defatted muscle proteins were hydrolyzed with 5% thermolysin. The resulting hydrolysate showed high inhibitory activity against angiotensin I-converting enzyme (inhibitory concentration(50) = 27.9 protein mug/mL) in vitro. A significant reduction of systolic blood pressure was observed when 500 and 2000 mg/kg of body weight were orally administered into spontaneously hypertensive rats. Angiotensin I-converting enzyme inhibitory peptides contained in the hydrolysate were isolated with various chromatographs. These 6 active peptides were Trp residue-containing dipeptides: Trp-Ala, Val-Trp, Trp-Met, Met-Trp, Ile-Trp, and Leu-Trp. The inhibitory concentration(50) values of these dipeptides ranged from 2.5 muM to 277.3 muM.
• Hosokawa, M., Hou, C.T., Weisleder, W. and Brown, W.:"Isolation and structure determination of 12,13,16-trihydroxy-9(Z)-octadecenoic acid produced from linoleic acid by Clavibacter sp. ALA2 and biosynthesis of tetrahydrofuranyl fatty acids", J. Am. Oi・・・

  2003 [Refereed][Not invited]
   

  Hosokawa, M., Hou, C.T., Weisleder, W. and Brown, W.:"Isolation and structure determination of 12,13,16-trihydroxy-9(Z)-octadecenoic acid produced from linoleic acid by Clavibacter sp. ALA2 and biosynthesis of tetrahydrofuranyl fatty acids", J. Am. Oil Chem. Soc., 80:145-149(2003)*
• Hosokawa, M., Hou, C.T. and Weisleder, D.:"Bioconversion products from n-3 and n-6 polyunsaturated fatty acids by Clavibacter sp. ALA2", J. Am. Oil. Chem. Soc., 80:1085-1091(2003)*

  2003 [Refereed][Not invited]
  
• Production of novel tetrahydroxyfuranyl fatty acids from alpha-linolenic acid by Clavibacter sp strain ALA2

  M Hosokawa, CT Hou, D Weisleder

  APPLIED AND ENVIRONMENTAL MICROBIOLOGY 69 (7) 3868 - 3873 0099-2240 2003 [Refereed][Not invited]
   

  Previously, it was reported that a newly isolated microbial culture, Clavibacter sp. strain ALA2, produced trillydroxy unsaturated fatty acids, diepxoy bicyclic fatty acids, and tetrabydroxyfuranyl fatty acids (THFAs) from linoleic acid (C. T. Hou, J. Am. Oil Chem. Soc. 73:1359-1362,1996; C. T. Hou. and R. J. Forman III, J. Ind. Microbiol. Biotechnol. 24:275-276, 2000; C. T. Hou, H. Gardner, and W. Brown, J. Am. Oil Chem. Soc. 75:14831487,1998; C. T. Hou, H. W. Gardner, and W. Brown, J. Am. Oil Chem. Soc. 78:1167-1169,2001). In this study, we found that Clavibacter sp. strain ALA2 produced novel THFAs, including 13,16-dihydroxy-12-THFA, 15epoxy-9(Z)-octadecenoic acid (13,16-dihydroxy-THFA), and 7,13,16-trihydroxy-12, 15-epoxy-9(Z)-octadecenoic acid (7,13,16-trihydroxy-THFA), from alpha-linolenic acid (9,12,15-octadecatrienoic acid). The chemical structures of these products were determined by gas chromatography-mass spectrometry and proton and C-13 nuclear magnetic resonance analyses. The optimum incubation temperature was 30degreesC for production of both hydroxy-THFAs. 13,16-Dihydroxy-THFA was detected after 2 days of incubation, and the concentration reached 45 mg/50 ml after 7 days of incubation; 7,13,16-trihydroxy-THFA was not detected after 2 days of incubation, but the concentration reached 9 mg/50 ml after 7 days of incubation. The total yield of both 13,16-dihydroxy-THFA and 7,13,16-trihydroxy-THFA was 67% (wt/wt) after 7 days of incubation at 30'C and 200 rpm. In previous studies, it was reported that Clavibacter sp. strain ALA2 oxidized the C-7, C-12, C-13, C-16, and C-17 positions of linoleic acid (n-6) into hydroxy groups. In this case, the bond between the C-16 and C-17 carbon atoms is saturated. In ot-linolenic acid (n-3), however, the bond between the C-16 and C-17 carbon atoms is unsaturated. It seems that enzymes of strain ALA2 oxidized the C-12-C-13 and C-16-C-17 double bonds into dihydroxy groups first and then converted them to hydroxy-THFAs.
• Dual action of isoprenols from herbal medicines on both PPAR gamma and PPAR alpha in 3T3-L1 adipocytes and HepG2 hepatocytes

  N Takahashi, T Kawada, T Goto, T Yamamoto, A Taimatsu, N Matsui, K Kimura, M Saito, M Hosokawa, K Miyashita, T Fushiki

  FEBS LETTERS 514 (2-3) 315 - 322 0014-5793 2002/03 [Refereed][Not invited]
   

  Several herbal medicines improve hyperlipidemia, diabetes and cardiovascular diseases. However, the molecular mechanism underlying this improvement has not yet been clarified. In this study, we found that several isoprenols, common components of herbal plants, activate human peroxisome proliferator-activated receptors (PPARs) as determined using the novel GAL4 ligand-binding domain chimera assay system with coactivator coexpression. Farnesol and geranylgeraniol that are typical isoprenols in herbs and fruits activated not only PPARgamma but also PPARalpha as determined using the chimera assay system. These compounds also activated full-length human PPARgamma and PPARalpha in CV1 cells. Moreover, these isoprenols upregulated the expression of some lipid metabolic target genes of PPARgamma and PPARalpha in 3T3-L1 adipocytes and HepG2 hepatocytes, respectively. These results suggest that herbal medicines containing isoprenols with dual action on both PPARgamma and PPARalpha can be of interest for the amelioration of lipid metabolic disorders associated with diabetes. (C) 2002 Federation of European Biochemical Societies. Published by Elsevier Science B.V. All rights reserved.
• Takahashi, N., Kawada, T., Gotoh, T., Yamamoto, T., Taimatsu, A., Matsui, N., Kimura, K., Saitoh, M. Hosokawa, M., Miyashita, K. and Fushiki, T. "Dual action of isoprenols from herbal medicines on both PPAR and PPAR g in 3T3-L1 adipocytes and HepG2 he・・・

  2002 [Refereed][Not invited]
   

  Takahashi, N., Kawada, T., Gotoh, T., Yamamoto, T., Taimatsu, A., Matsui, N., Kimura, K., Saitoh, M. Hosokawa, M., Miyashita, K. and Fushiki, T. "Dual action of isoprenols from herbal medicines on both PPAR and PPAR g in 3T3-L1 adipocytes and HepG2 hepatocytes". FEBS Lett., 514: 315-322(2002) *
• Proteolytic conversion of squid hepatopancreas and upstream chum salmon muscle to angiotensin I-converting enzyme inhibitory peptides

  S Ono, M Hosokawa, A Inoue, D Yamadda, K Takahashi

  NIPPON SUISAN GAKKAISHI 公益社団法人日本水産学会 68 (2) 192 - 196 0021-5392 2002 [Refereed][Not invited]
   

  We tried to convert the upstream chum salmon muscle and squid hepatopancreas, which are major marine by-products in northern Japan, to angiotensin I-converting enzyme (ACE) inhibitory peptides. Hydrolysates prepared from defatted chum salmon muscle and squid liver with some protease showed the inhibitory activity against ACE. When deflated chum salmon muscle was hydrolyzed for 2 h at 37degreesC with 5% thermolysin, the inhibitory activity reached 73.0% at 78.9 mug/mL. The yield of the hydrolysate was 86.9%.
• Dietary conjugated linolenic acid inhibits azoxymethane-induced colonic aberrant crypt foci in rats

  H Kohno, R Suzuki, R Noguchi, M Hosokawa, K Miyashita, T Tanaka

  JAPANESE JOURNAL OF CANCER RESEARCH 93 (2) 133 - 142 0910-5050 2002 [Refereed][Not invited]
   

  The modifying effects of dietary feeding of conjugated linolenic acid (CLN) isolated from the seeds of bitter gourd (Momordica charantia) on the development of azoxymethane (AOM)-induced colonic aberrant crypt foci (ACF) were investigated in male F344 rats to predict its possible cancer chemopreventive efficacy. The effect of CLN on the proliferating cell nuclear antigen (PCNA) index in colonic ACF was also examined. Rats were given subcutaneous injections of AOM (20 mg/kg body weight) once a week for 2 weeks to induce ACE They also received the experimental diet containing 0.01%, 0.1% or 1% CLN for 5 weeks, starting one week before the first dosing of AOM. AOM exposure produced a substantial number of ACF (108 +/- 21/rat) at the end of the study (week 4). Dietary administration of CLN caused a significant reduction in the frequency of ACF: 87 +/- 14 (19.4% reduction, P<0.05) at a dose of 0.01%, 69 +/- 28 (36.1% reduction, P<0.01) at a dose of 0.1% and 40 +/- 6 (63.0% reduction, P<0.001) at a dose of 1%. Also, CLN administration lowered the PCNA index and induced apoptosis in ACE These findings might suggest possible chemopreventive activity of CLN in the early phase of colon tumorigenesis through modulation of cryptal cell proliferation activity and/or apoptosis.
• Liposomal sn-2 DHA inserted phospholipid decreases tumor size of fibrosarcoma

  Akihito Fujimoto, Jin-Ichi Sasaki, Masashi Hosokawa, Koretaro Takahashi

  Fisheries Science 68 1639 - 1640 1444-2906 2002 [Refereed][Not invited]
  
• Dietary effects of bitter gourd oil on blood and liver lipids of rats

  R Noguchi, Y Yasui, R Suzuki, M Hosokawa, K Fukunaga, K Miyashita

  ARCHIVES OF BIOCHEMISTRY AND BIOPHYSICS 396 (2) 207 - 212 0003-9861 2001/12 [Refereed][Not invited]
   

  Bitter gourd is widely used as an edible plant in Asia. In this study, we evaluated the effects of bitter gourd oil (BGO) on the blood and liver lipids of rats. Three groups of rats were given a basal diet (AIN-93G) containing 7% fat by weight. The dietary fat consisted of soybean oil (control), soybean oil + BGO (6.5:0.5, w/w; 0.5% BGO), or soybean oil + BGO (5:2, w/w; 2.0% BGO). This fat treatment gave 3.4 and 15.4% of cis(c)9,trans(t)11,t13-18:3 in the dietary fat of 0.5 and 2.0% BGO, respectively. Fatty acid analysis showed the occurrence of c9,t11-18:2 in the liver of rats fed BGO diets, whereas this conjugated linoleic acid (CLA) isomer was not detected in the liver of rats fed the control diet. Furthermore, dietary BGO decreased the concentration of 18:2n-6 and increased the concentration of 22:6n-3. The formation of the CLA isomer in the liver lipids of rats fed BGO diets could be explained by either of the following two metabolic pathways, namely, enzymatic biohydrogenation of c9,t11,t13-18:3 or enzymatic isomerization of c9,c12-18:2. The BGO diets had significantly reduced free cholesterol levels with a trend toward an increase in HDL cholesterol, but there was no significant change in the total cholesterol. The dietary BGO also affected the level of plasma hydroperoxides. A slight but significant increase in hydroperoxides was found in the rats fed 2.0% BGO. This may be attributed to the lower oxidative stability of c9,t11,t13-18:3 in BGO. (C) 2001 Elsevier Science.
• Ligands for peroxisome proliferator-activated receptors alpha and gamma inhibit chemically induced colitis and formation of aberrant crypt foci in rats

  T Tanaka, H Kohno, S Yoshitani, S Takashima, A Okumura, A Murakami, M Hosokawa

  CANCER RESEARCH 61 (6) 2424 - 2428 0008-5472 2001/03 [Refereed][Not invited]
   

  The biological role of the peroxisome proliferator-activated receptors (PPARs) in various diseases, including inflammation and cancer, has been highlighted recently. Although PPAR gamma ligands have been found to inhibit mammary carcinogenesis in rodents, the effects on colon tumorigenesis are controversial. In the present study, three different experiments were conducted to investigate the modifying effects of PPARs ligands (PPAR alpha and PPAR gamma) on colitis and an early phase of colitis-related colon carcinogenesis in male F344 rats. In the first experiment, gastric gavage of troglitazone (PPAR gamma ligand, 10 or 100 mg/kg body weight) or bezafibrate (PPAR alpha ligand, 10 or 100 mg/kg body weight) inhibited colitis induced by dextran sodium sulfate (DSS) and lowered trefoil factor-2 content in colonic mucosa. In the second experiment, dietary administration (0.01 or 0.05% in diet) of troglitazone and bezafibrate for 4 weeks significantly reduced azoxymethane (AOM, two weekly s.c. injections, 20 mg/kg body weight)-induced formation of aberrant crypts foci, which are precursor lesions for colon carcinoma. In the third experiment, dietary administration (0.01% in diet for 6 weeks) of pioglitazone (PPAR gamma ligand), troglitazone, and bezafibrate effectively suppressed DSS/AOM-induced ACF. Administration of both ligands significantly reduced cell proliferation activity in colonic mucosa exposed to DSS and AOM. Our results suggest that synthetic PPARs ligands (PPAR alpha and PPAR gamma) can inhibit the early stages of colon tumorigenesis with or without colitis.
• Kohno, H., Yoshitani, S. Takashima, S., Okumura, A., Hosokawa, M. and Yamaguchi, M., Tanaka, T. : “Troglitazone, a ligand for peroxisome proliferator-activated receptor γ, inhibits chemically-induced aberrant cript foci in rats”, Jpn. J. Cancer Res. 92・・・

  2001 [Refereed][Not invited]
   

  Kohno, H., Yoshitani, S. Takashima, S., Okumura, A., Hosokawa, M. and Yamaguchi, M., Tanaka, T. : “Troglitazone, a ligand for peroxisome proliferator-activated receptor γ, inhibits chemically-induced aberrant cript foci in rats”, Jpn. J. Cancer Res. 92: 396-403 (2001)
• Synergistic effects of highly unsaturated fatty acid-containing phosphatidylethanolamine on differentiation of human leukemia HL-60 cells by dibutyryl cyclic adenosine monophosphate

  M Hosokawa, A Sato, H Ishigamori, H Kohno, T Tanaka, K Takahashi

  JAPANESE JOURNAL OF CANCER RESEARCH 92 (6) 666 - 672 0910-5050 2001 [Refereed][Not invited]
   

  Highly unsaturated fatty acid-containing phospholipid (HUFA-PL) has many nutritional and medical applications. We investigated the effect of HUFA-PL on differentiation of human leukemia HL-60 cells induced by dibutyryl cyclic adenosine monophosphate (dbcAMP). HUFA-containing phosphatidylethanolamine (HUFA-PE), such as salmon testis PE, significantly enhanced dbcAMP-induced cell differentiation. A combined treatment of 200 muM dbcAMP with 50 muM HUFA-PE increased the nitroblue tetrazolium (NBT)-reducing activity, which is an indicator of differentiation, to a level comparable to that in the case of 500 muM dbcAMP treatment. In contrast, HUFA-lyso PE (a monoacyl form) did not exert an enhancing effect on dbcAMP-induced differentiation. The enhancing effect of HUFA-PE was suppressed by a protein kinase C inhibitor, staurosporine, while a protein kinase A inhibitor, H.8, did not suppress the enhancing effect. These findings suggest that HUFA-PE might enhance dbcAMP-induced differentiation through modulation of the protein kinase C signaling pathway in HL-60 cells.
• Noguchi, R., Yasui, Y., Suzuki, R., Hosokawa, M., Fukunaga, K. and Miyashita, K.: “Dietary effect of bitter gourd oil on blood and liver lipids of rats”, Arch. Biochem. Biophys, 396: 207-212 (2001)

  2001 [Refereed][Not invited]
  
• Troglitazone, a ligand for peroxisome proliferator-activated receptor γ, inhibits chemically-induced aberrant crypt foci in rats

  Hiroyuki Kohno, Shin-Ichiro Yoshitani, Shigeki Takashima, Ataru Okumura, Masashi Hosokawa, Nobuo Yamaguchi, Takuji Tanaka

  Japanese Journal of Cancer Research 92 (4) 396 - 403 0910-5050 2001 [Refereed][Not invited]
   

  The biological roles of peroxisome proliferator-activated receptors (PPARs) in various diseases, including inflammation and cancer, have been highlighted recently. Although PPARγ ligand is suspected to play an important role in carcinogenesis, its effects on colon tumorigenesis remain undetermined. The present time-course study was conducted to investigate possible modifying effects of a PPARγ ligand, troglitazone, on the development and growth of aberrant crypt foci (ACF), putative precursor lesions for colon carcinoma, induced by azoxymethane (AOM) or dextran sodium sulfate (DSS) in male F344 rats. Oral troglitazone (10 or 30 mg/kg body weight (b.w.)) significantly reduced AOM (two weekly subcutaneous injections, 20 mg/kg b.w.)-induced ACF. Treatment with troglitazone increased apoptosis and decreased polyamine content and ornithine decarboxylase (ODC) activity in the colonic mucosa of rats treated with AOM. Gastric gavage of troglitazone also inhibited colitis and ACF induced by DSS (1% in drinking water), in conjunction with increased apoptosis and reduced colonic mucosal polyamine level and ODC activity. Our results suggest that troglitazone, a synthetic PPARγ ligand, can inhibit the early stage of colon tumorigenesis with or without colitis.
• Production of tailor-made polyunsaturated phospholipids through bioconversions

  K Takahashi, M Hosokawa

  JOURNAL OF LIPOSOME RESEARCH 11 (4) 343 - 353 0898-2104 2001 [Refereed][Not invited]
  
• Hosokawa,M.,Shimatani,T.,Kaneda,T.,Inoue Y.and Takahashi K.:“Coversion to Docosahexaenoic Acid-Containing Phosphatidylserine from Squid Skin Lecithin by Phospholipase D-Mediated Transphosphatidylation”,Journal of Agricultural and Food Chemistry, 48:455・・・

  2000 [Refereed][Not invited]
   

  Hosokawa,M.,Shimatani,T.,Kaneda,T.,Inoue Y.and Takahashi K.:“Coversion to Docosahexaenoic Acid-Containing Phosphatidylserine from Squid Skin Lecithin by Phospholipase D-Mediated Transphosphatidylation”,Journal of Agricultural and Food Chemistry, 48:4550-4554(2000)※
• Antibacterial activity against cariogenic bacteria and inhibition of insoluble glucan production by free fatty acids obtained from dried Gloiopeltis furcata

  H Kurihara, Y Goto, H Aida, M Hosokawa, K Takahashi

  FISHERIES SCIENCE 65 (1) 129 - 132 0919-9268 1999/02 [Refereed][Not invited]
   

  Antibacterial substance for mutans streptococci was obtained from dried Gloiopeltis furcata by chromatographic separation. The substance was determined to be a mixture of free fatty acids (FFAs), from the results of instrumental analyses. The FFAs obtained showed moderate antibacterial activity against mutans streptococci, with minimal inhibitory concentration values between 25 and 50 mu g/ml. These FFAs also inhibited insoluble glucan production catalyzed by glucosyltransferase of Streptococcus sobrinus. Experiments carried out by using commercially available fatty acids indicated that unsaturated fatty acids showed more potent inhibition against insoluble glucan production than saturated fatty acids.
• Lipase-mediated Interesterification of a Highly Unsaturated Fatty Acid-containing Acylglycerol Mixture

  HOSOKAWA Masashi, HORII Aki, TAKAHASHI Koretaro

  Journal of Japan Oil Chemists' Society 日本油化学会 = Japan Oil Chemists' Society 48 (1) 51 - 53 1341-8327 1999/01/20 [Refereed][Not invited]
   

  The triacylglycerol (TG) molar ratio was augmented by Lipozyme IM-mediated interesterification of docosahexaenoic acid (DHA) rich (72.0%) partial acylglycerol mixtures (DHA-70G). DHA-70G used as substrate contained TG at 49.3%. Through interesterification mediated by Lipozyme IM previously dried with phosphoric anhydride prior to reaction, TG increased to 64.5% by 24h.The addition of molecular sieves in the reaction system at this point was effective by further increased TG in the reaction product. This content finally reached 81.0% at 48h interesterification. DHA content dominated for 73.7...
• Toda,H.,Kurihara,H.,Hosokawa,M. and Takahashi,K.:"Inhibition of α-glucuronidase by purine nucleotides",Fisheries Sci.,65:667-668(1999)*

  1999 [Refereed][Not invited]
  
• Hosokawa,M.,Wanezaki,S.,Miyauchi,K.,Kurihara,H.,Kohno,H.,Kawabata,J.,Odashima, S, and Takahashi,K.:"Apoptosis-inducing effect of fucoxanthin on human leukemia cell line HL-60",Food Sci.Technol.Res.,5:243-246(1999)*

  1999 [Refereed][Not invited]
  
• Differentiation-and Apoptosis-inducing Activities of Phospholipids Containing Docosahexaenoic Acid for Mouse Myeloid Leukemia M1 Cells

  HOSOKAWA Masashi, MINAMI Keiichi, KOHNO Hiroyuki, TANAKA Yukihisa, HIBINO Hidehiko

  Fisheries science : FS 65 (5) 798 - 799 0919-9268 1999 [Refereed][Not invited]
  
• Inhibition of β-Glucuronidase by Purine Nucleotides

  Hiroyuki Tohda, Hideyuki Kurihara, Masashi Hosokawa, Koretaro Takahashi

  Fisheries science 65 (4) 667 - 668 0919-9268 1999 [Refereed]
  
• Apoptosis-Inducing Effect of Fucoxanthin on Human Leukemia Cell Line HL-60.

  Masashi HOSOKAWA, Satoshi WANEZAKI, Kozue MIYAUCHI, Hideyuki KURIHARA, Hiroyuki KOHNO, Jun KAWABATA, Shizuo ODASHIMA, Koretaro TAKAHASHI

  Food Science and Technology Research 5 (3) 243 - 246 1344-6606 1999 [Refereed]
  
• Preparation of highly unsaturated fatty acid-containing phosphatidylcholine by transesterification with phospholipase A(2)

  M Hosokawa, M Ito, K Takahashi

  BIOTECHNOLOGY TECHNIQUES 12 (8) 583 - 586 0951-208X 1998/08 [Refereed][Not invited]
   

  To prepare phospholipids containing highly unsaturated fatty acids (HUFA) by a one-step enzymatic reaction, 14 mu mol soy phosphatidylcholine (soy PC) in combination with 60 mu mol HUFA as acyl donor were transesterified with porcine pancreatic phospholipase A(2). Specify HUFA was successively incorporated into the sn-2 position of soy PC by this reaction at greater than 70% of total fatty acids at sn-2.
• Hosokawa,M.,Ito,M and Takahashi,K.:"Preparation of highly unsaturated fatty acid-containing phosphatidylcholine by transesterification with phospholipase A2",Biotechnol.Tech.,12:583-586(1998)*

  1998 [Refereed][Not invited]
  
• Increase in Deformability of Human Erythrocytes through the Action of β-Lysophospholipid Rich in n-3 Polyunsaturated Fatty Acid Content

  HOSOKAWA Masashi, ONO Masayo, TAKAHASHI Koretaro, INOUE Yoshikazu

  Journal of Japan Oil Chemists' Society Japan Oil Chemists' Society 47 (12) 1313 - 1318,1360 1341-8327 1998 [Refereed][Not invited]
   

  Lysophosphatidylcholine (LPC) that binds to docosahexaenoic acid (DHA) at the <I>sn</I>-2 position (β-DHA-LPC) was prepared by the Lipozyme mediated selective partial hydrolysis of DHA-enriched egg yolk lecithin. The β-DHA-LPC was found to be rapidly incorporated into erythrocyte cells and increase their deformability at 5μM/erythrocyte.
• Water activity-adjusted enzymatic partial hydrolysis of phospholipids to concentrate polyunsaturated fatty acids

  M Ono, M Hosokawa, Y Inoue, K Takahashi

  JOURNAL OF THE AMERICAN OIL CHEMISTS SOCIETY 74 (11) 1415 - 1417 0003-021X 1997/11 [Refereed][Not invited]
   

  Selective partial hydrolyses of egg yolk phospholipid and squid skin phospholipid were carried out. By keeping the water activity (a(W)) of Lipozyme IM at an intermediate level, it was easy to concentrate docosahexaenoic acid (DHA). II was also possible to concentrate both DHA and arachidonic acid (AA) simultaneously to a certain level under this a(W) range. However, it was impossible to concentrate AA alone when DHA was present. Though there is a limitation in concentrating AA exclusively, the proposed a(W)-adjusted hydrolytic reaction is a promising way for preparing phospholipids rich in DHA.
• Effects of Phospholipids Containing Docosahexaenoic Acid on Differentiation and Growth of HL-60 Human Promyelocytic Leukemia Cells

  TOCHIZAWA Kaoru, HOSOKAWA Masashi, KURIHARA Hideyuki, KOHNO Hiroyuki, ODASHIMA Shizuo, TAKAHASHI Koretaro

  Journal of Japan Oil Chemists' Society Japan Oil Chemists' Society 46 (4) 383 - 390,457 1341-8327 1997 [Refereed][Not invited]
   

  Effects of docosahexaenoic acid (DHA) -containing phospholipids (DHA-PLs) on retionic acid (RA) -induced cell differentiation and growth inhibition of the human promyelocytic leukemia cell line, HL-60 were investigated. Cell differentiation judged from NBT reduction assay reached to 5060% by incubation with 12.5μM DHA-containing phosphatidylcholines (DHA-PCs) prior to the addition of RA. Differentiation without DHA-PCs remained at 39.2%. By prior incubation with DHA-containing phosphatidylethanolamines (DHA-PEs), differentiation was 70% at 12.5μM. DHA-PEs induced NBT reducing activity to as much as 2030%, and inhibited cell growth in the absence of RA. Activity for promoting cell differentiation differed according to DHA-PL molecular species. 1-Palmitoyl (or oleoyl)- 2-docosahexaenoyl-<I>sn</I>-glycero-3-phosphocholine (or phosphoethanolamine) was found most effective molecular species for enhancing RA-induced cell differentiation.
• Ono, M., Hosokawa, M., Inoue, Y. and Takahashi, K. : "Water Activity Adjusted Enzymatic Partial Hydrolysis of Phospholipids to Concentrate Polyunsaturated Fatty Acids", J. Am. Oil Chem. Soc., 74 : 1415-1417 (1997)＊

  1997 [Refereed][Not invited]
  
• Ono, M., Hosokawa, M., Inoue, Y. and Takahashi, K. : "Concentration of Docosahexaenoic Acid-Containing Phospholipids through Lipozyme IM-Mediated Hydrolysis", J. Jpn. Oil Chem. Soc., 46 : 867-872 (1997)＊

  1997 [Refereed][Not invited]
  
• PREPARATION OF THERAPEUTIC PHOSPHOLIPIDS THROUGH PORCINE PANCREATIC PHOSPHOLIPASE A(2)-MEDIATED ESTERIFICATION AND LIPOZYME-MEDIATED ACIDOLYSIS

  M HOSOKAWA, K TAKAHASHI, Y KIKUCHI, M HATANO

  JOURNAL OF THE AMERICAN OIL CHEMISTS SOCIETY 72 (11) 1287 - 1291 0003-021X 1995/11 [Refereed][Not invited]
   

  Highly unsaturated fatty acid-containing phospholipids (HUFA-PL), which serve to increase the deformability of human red blood cells, were prepared through porcine phospholipase A(2)-mediated esterification of the lysophosphatidylcholone, which was derived from soy phosphatidylcholine (PC), and by Lipozyme-mediated acidolysis. Through these processes, phospholipase A(2), with formamide as a water mimic, enhanced the incorporation of HUFA into position sn-2 of PC and suppressed hydrolysis of the synthesized PL. On the other hand, Lipozyme-mediated acidolysis between position sn-1 of soy PC and HUFA was enhanced by a combination of water and propylene glycol. Simultaneously, the recovered PL products showed decreased hydrolysis of newly synthesized health-beneficial HUFA-PL.
• APPLICATION OF WATER MIMICS ON PREPARATION OF EICOSAPENTAENOIC AND DOCOSAHEXAENOIC ACIDS CONTAINING GLYCEROLIPIDS

  M HOSOKAWA, K TAKAHASHI, N MIYAZAKI, K OKAMURA, M HATANO

  JOURNAL OF THE AMERICAN OIL CHEMISTS SOCIETY 72 (4) 421 - 425 0003-021X 1995/04 [Refereed][Not invited]
   

  To obtain enhanced incorporation of highly unsaturated fatty acids and recovery of glycerolipid products, organic solvents with high dielectric constants (water mimics) were substituted for part of the essential water for lipase activation to study their effect on acidolysis and transesterification. In acidolysis/transesterification of fish oil triglycerides and eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA), Lipozyme IM-60 with ethylene glycol as a water mimic enhanced the incorporation of EPA and suppressed the hydrolysis of synthesized glycerolipid. On the other hand, transesterification between soy phosphatidylcholine and EPA was enhanced by a water and propylene glycol combination. In a nonaqueous medium that contained appropriate amounts of water and organic solvents (water mimics), Lipozyme IM-60 increased transesterification of EPA into soy phosphatidylcholine. Simultaneously, the recovered glycerolipid products showed decreased hydrolysis of newly synthesized EPA- and DHA-containing glycerolipids.
• EFFECT OF IN-VITRO TREATMENT WITH GLYCEROPHOSPHOLIPID CONTAINING HIGHLY POLYUNSATURATED FATTY-ACIDS ON DEFORMABILITY OF HUMAN ERYTHROCYTES

  M NOJIMA, M HOSOKAWA, K TAKAHASHI, M HATANO, Y KIKUCHI

  NIPPON SUISAN GAKKAISHI 日本水産學會 61 (2) 197 - 203 0021-5392 1995/03 [Refereed][Not invited]
   

  The flow rate of human erythrocytes on microchannels was measured to evaluate their deformability when treated with soy phospholipids, hydrogenated soy phospholipids, phospholipids containing icosapentaenoic acid (EPA) and docosahexaenoic acid (DHA). EPA-containing phosphatidylserine and DHA-containing phosphatidylcholine or phosphatidylethanolamine were observed to be the most effective lipid molecular species to improve the deformability of human erythrocytes, while hydrogenated phospholipids tended to impair deformability.
• EFFECT OF EPA AND DHA CONTAINING GLYCEROPHOSPHOLIPID MOLECULAR-SPECIES ON THE FLUIDITY OF ERYTHROCYTE CELL-MEMBRANES

  M NOJIMA, M HOSOKAWA, K TAKAHASHI, M HATANO

  FISHERIES SCIENCE 60 (6) 729 - 734 0919-9268 1994/12 [Refereed][Not invited]
   

  Fluorescence depolarization of erythrocytes was measured to evaluate the increase in fluidity of cell membrane due to treatment with soy phospholipids, hydrogenated soy phospholipids, icosapentaenoic acid (EPA), and docosahexaenoic acid (DHA) containing phospholipids. When the erythrocytes were treated with EPA or DHA containing phosphatidylethanolamine or phosphatidylserine, the least fluorescence polarization was observed, followed by EPA or DHA containing phosphatidylcholine. All of the hydrogenated phospholipids increased fluorescence depolarization. 2,4,6-Trinitrobenzenesulfonic acid fluorescence quenching proved that the phospholipids used for erythrocytes treatment are incorporated mainly into the inner layer of their cell membrane.
• Phospholipase A2‐mediated synthesis of phosphatidylethanolamine containing highly unsaturated fatty acids

  MASASHI HOSOKAWA, KORETARO TAKAHASHI, MUTSUO HATANO, MAKOTO EGI

  International Journal of Food Science & Technology 29 (6) 721 - 725 1365-2621 1994 [Refereed][Not invited]
   

  Phosphatidylethanolamine (PE), which contains highly unsaturated fatty acid (HUFA) specifically in sn‐2 position, was synthesized by porcine pancreatic phospholipase A2‐catalysed esterification of lysophosphatidylethanolamine with HUFA. the use of glycerol containing only a little water as solvent was the important key to this reaction. A 27% yield of up to 94.5%sn‐2 eicosapentaenoic acid (EPA)‐containing PE was obtained using > 99% EPA as a substrate HUFA. Copyright © 1994, Wiley Blackwell. All rights reserved
• SYNTHESIS OF PHOSPHATIDYLCHOLINE CONTAINING HIGHLY UNSATURATED FATTY-ACID BY PHOSPHOLIPASE-A2 AND EFFECT ON RETINOIC ACID-INDUCED DIFFERENTIATION OF HL-60 CELLS

  M HOSOKAWA, H OHSHIMA, H KOHNO, K TAKAHASHI, M HATANO, S ODASHIMA

  NIPPON SUISAN GAKKAISHI 日本水産學會 59 (2) 309 - 314 0021-5392 1993/02 [Refereed][Not invited]
   

  Phosphatidylcholine containing highly unsaturated fatty acid (HUFA-PC) was prepared by porcine pancreatic phospholipase A2, which catalyzed esterification between lysophosphatidylcholine (LPC) and highly unsaturated fatty acid (HUFA), under a scaled-up reaction system. Fatty acid mixture prepared from sardine oil, purified eicosapentaenoic acid (EPA), and purified docosahexaenoic acid (DHA) were used as the substrates of HUFA. The yield of HUFA-PC was 17.0-19.9%. Synthesized phosphatidylcholine containing DHA (DHA-PC) prepared from LPC and DHA had a DHA content of 46.5%, which indicated almost the maximum theoretical incorporation rate. DHA-PC molecular species consisted of (16 : 0, DHA), (18 : 0, DHA), (18 : 1, DHA), (18 : 2, DHA), and (18 : 3, DHA). It was obvious that the acyl groups in sn-1 of these molecular species originated from the substrate LPC. The rate of retinoic acid induced differentiation of the human promyelocytic leukemia cell line (HL-60) was promoted by growing HL-60 cells in 10% FBS-PNM pre-treated with either EPA-PC or DHA-PC.
• Polyunsaturated Fatty Phosphatide Synthesis by Industrial Phospholipase A2

  HOSOKAWA Masashi, TAKAHASHI Koretaro, HATANO Mutsuo, EGI Makoto

  Journal of the Japanese Society for Food Science and Technology 日本食品工業学会 38 (8) p695 - 698 0029-0394 1991/08 [Refereed][Not invited]
   

  Phospholipids consist of eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) were synthesized by use of an industrial pancreatic phospholipase A₂. Lysophosphatidylcholine (LPC) isolated from a commercial emulsifier, mixed fatty acids obtained from sardine oil, the concentrated sardine oil (EPA+DHA>60%), and 99.4% EPA were used as substrates. Optimum water content was found to 13.4%/system, and optimum ratio of LPC to mixed free fatty acids was 11:18 in weight base. When the substrate contained free EPA+DHA over 68%, the obtained phosphatidylcholine (PC) contained more than 31% of EPA+DHA. And when the substrate was 99.4% EPA, the obtained PC contained more than 45% of EPA, indicating that more than 90% of EPA was incorporated into sn-2 position of the PC molecule.
• MOLECULAR-SPECIES OF THE ENZYMATICALLY SYNTHESIZED POLYUNSATURATED FATTY-ACID RICH TRIGLYCERIDE

  K OSADA, K TAKAHASHI, M HATANO, M HOSOKAWA

  NIPPON SUISAN GAKKAISHI 日本水産学会 57 (1) 119 - 125 0021-5392 1991 [Refereed][Not invited]
   

  Enzymatic glyceride synthesis and acid interchange using icosapentaenoic acid (EPA), docosahexaenoic acid (DHA), gamma-linolenic acid (GLA), and EPA and DHA concentrated saponified fatty acid mixture obtained from sardine oil were carried out through the use of four kinds of microbial lipases. Lipase TOYO (Chromobacterium viscosum) was the most effective enzyme for glyceride synthesis as well as acid interchange of triglyceride (TG) rich in polyunsaturated fatty acids. There was a general tendency, except with Lipase TOYO, that DHA was difficult to be incorporated into a glycerol molecule compared to the other fatty acids. Fatty acid composition of TG synthesized from a highly concentrated fatty acid mixture (EPA+DHA: 68.8%) by the use of Lipase TOYO and Lipase OF (Candida cylindracea), directly reflected that of the substrate fatty acid flixture (i.e. EPA+DHA was more than 65%). And the TG molecular species having combination of only EPA and DHA such as (EPA, EPA, EPA), (EPA, EPA, DHA), (EPA, DHA, DHA), and (DHA, DHA, DHA) were dominant accounting for the half of the obtained TG.

Books etc

• 生体におけるリン脂質の性状と機能性

  細川, 雅史, 菅原, 達也, 機能油脂懇話会 
  建帛社 2020/10 (ISBN: 9784767962153) vi, 141p
• 水産生物機能性成分の研究論文概要集 : 水産生物機能性成分と研究動向

  細川, 雅史 
  日本水産油脂協会 2016/07 vi, 50p
• 「第7章2 カロテノイドの吸収と代謝」、宮澤陽夫編、「食品機能性成分の吸収・代謝機構」

  細川雅史 
  シーエムシー出版、東京 2013 p215-223
• Chapter 15 Chemopreventive effects of astaxanthin on inflammatory bowel disease and inflammation-related colon carcinogenesis. Editor, O Sommerburg, W Siems, K Kraemer . “Carotenoids and vitamin A in translational medicine”.

  M. Hosokawa, Y. Yasui 
  Boca Raton, USA.. 2013 p293-308.
• 第6章1-7 脂溶性色素、日本油化学会編、「油脂・脂質・界面活性剤データブック「I編 油脂・脂質」

  細川雅史 
  丸善、東京 2012 p228-234
• Omega-3 oils : applications in functional foods

  Hernandez Ernesto M, 細川 雅史 
  AOCS Press 2011
• Omega-3 oils : applications in functional foods

  Hernandez, Ernesto M., 細川, 雅史 
  AOCS Press 2011 (ISBN: 9781893997820) v, 305 p.
• 褐藻脂質中に含まれるフコキサンチンの抗肥満および抗糖尿病作用

  細川, 雅史 
  日本水産油脂協会 2010 21p
• The Encyclopedia of Industrial Biotechnology

  John Wiley & Sons, Inc. 2010
• Biocatalysis and Biomolecular Engineering

  John Wiley & Sons, Inc. 2010
• Handobook of seafood quality, safety and health application.

  Wiley-Blackwell 2010
• 油脂・脂質の基礎と応用−栄養・健康から工業まで−（改定第2版）

  日本油化学会 2009
• Nutrigenomics and proteomics in health and disease

  Wiley-Blackwell 2009
• 油脂・脂質の基礎と応用－栄養・健康から工業まで－（改定第2版）

  日本油化学会 2009
• カロテノイドの科学と最新応用技術

  シーエムシー出版 2009
• Biocatalysis and Agricultural Biotechnology

  CRC Press Taylor & Francis 2009
• 水産物由来の脂溶性成分による脂肪細胞のライフサイクル制御機能の解明

  細川, 雅史 
  [北海道大学] 2008/04 1冊
• Composition, functionality and potential applications of seaweed lipids.

  John Wiley & Sons, Inc 2008
• Enzymatic production of marine-derived protein hydrolysates and their bioactive peptides for use in foods and nutraceuticals.

  John Wiley & Sons, Inc 2008
• Antiobesity effect of fucoxanthin from edible seaweeds and its multibiological functions.

  ACS 2008
• Cancer: Disease Progression and Chemoprevention

  Research Signpost 2007
• Biocatalysis and biotechnology for functional foods and industrial products

  CRC Press Taylor & Francis 2007
• Marine Nutraceuticals and Functional Foods

  CRC Press Taylor & Francis 2007
• Cancer: Disease Progression and Chemoprevention

  Research Signpost 2007
• Biocatalysis and biotechnology for functional foods and industrial products

  CRC Press Taylor & Francis 2007
• Nutraceutical and Specialty Lipids and their Co-Products

  CRC Press Taylor & Francis 2006
• Nutraceutical and Specialty Lipids and their Co-Products

  CRC Press Taylor & Francis 2006

MISC

• 高度不飽和ホスファチジルグリセロールの酵素的合成と肝臓脂質低下作用

  陳莉萍, 高谷直己, 別府文章, 宮下和夫, 細川雅史  日本水産学会大会講演要旨集  2020-  2020
• 非アルコール性脂肪肝炎誘導モデルマウスに対する海洋性カロテノイドfucoxanthinの炎症抑制作用

  高谷直己, 河野友香, 別府史章, 宮下和夫, 細川雅史  日本栄養・食糧学会大会講演要旨集  74th-  2020
• フコキサンチンによる非アルコール性脂肪肝炎抑制効果 (特集 脂質がつなぐもの : 生物工学的脂質研究が導く新しい学際研究と産業(後編))

  高谷 直己, 細川 雅史  生物工学会誌  98-  (10)  529  -532  2020
• Furan fatty acidss-a new approach

  Matt Miller, Donato Romanazzi, Masashi Hosokawa, Hajime Uchida  INFORM  30-  25  -27  2019/03/01  [Not refereed][Not invited]
   

  © 2019, American Oil Chemists' Society. All rights reserved. Furan fatty acids (F-acids) are minor lipid components in organisms, plants, and mammals. F-acids have a number of different species according to chain length on the alpha-positions of the furan ring and methylation of the ring, as outlined in Figure 1. F-acids have been identified in a number of fish species, particularly in the liver and gonads. The concentration of F-acids in liver oil from Northern pike can be as high as 5%, but the F-acids in oil from the pike’s testes can be as high as 65%.
• 海洋性カロテノイドfucoxanthinの新規生体内開裂物による炎症抑制機構

  高谷直己, 田谷大輔, 宮下和夫, 細川雅史  日本栄養・食糧学会大会講演要旨集  73rd-  2019
• アスタキサンチン由来アポカロテノイドによる炎症抑制機構

  高谷直己, 別府史章, 宮下和夫, 細川雅史  日本油化学会年会講演要旨集  58th-  2019
• 食品中のカロテノイド類

  小泉次郎, 細川雅史  農業および園芸  92-  (10)  875‐880  -880  2017/10/01  [Not refereed][Not invited]
  
• Fucoxanthin in the management of obesity and its related disorders

  Kazuo Miyashita, Masashi Hosokawa  JOURNAL OF FUNCTIONAL FOODS  36-  195  -202  2017/09  [Not refereed][Not invited]
   

  Interest in a novel biological agent fucoxanthin (Fx) has been growing in recent years, which connects Fx to increases in energy expenditure and has implications for the management of obesity and type 2 diabetes mellitus (T2DM). Several molecular targets offer the most hope for influencing energy expenditure. One of the keys for success will be uncoupling protein 1 (UCP1). Fx can reduce the excess fat accumulation in abdominal white adipose tissue (WAT), which is primarily based on the up-regulation of the thermogenesis through UCP1 expression in both brown adipose tissue (BAT) and abdominal WAT. Fx down regulates the expression of pro-inflammatory adipokines involved in insulin resistance from abdominal WAT and subsequently normalizes the blood glucose level. In addition, Fx can promote glucose utilization in skeletal muscle by up-regulating glucose transporter 4 (GLUT4) expression and promoting its translocation to the cell membrane from the cytosol. (C) 2017 Elsevier Ltd. All rights reserved.
• Structure and functions of marine lipids

  細川 雅史  Food style 21  21-  (6)  39  -44  2017/06  [Not refereed][Not invited]
  
• 食品因子による生活習慣病予防・改善機構の解明をめざして―3 褐藻由来フコキサンチンの抗肥満・抗糖尿病効果とその機序

  西川翔, 細川雅史, 宮下和夫  化学と生物  54-  (8)  580‐585  -585  2016/07/20  [Not refereed][Not invited]
  
• 海洋性カロテノイドの健康機能

  細川 雅史  ニューフードインダストリー  58-  (6)  7  -12  2016/06  [Not refereed][Not invited]
  
• Biological importance of fucoxanthin and its metabolites

  Kazuo Miyashita, Masashi Hosokawa, Nana Mikami, Yasuo Kokai  ABSTRACTS OF PAPERS OF THE AMERICAN CHEMICAL SOCIETY  250-  2015/08  [Refereed][Not invited]
  
• 海洋細菌Jejuia pallidilutea 11shimoA1株の新規カロテノイド生合成系解析

  高谷直己, 澤辺智雄, 宮下和夫, 細川雅史  マリンバイオテクノロジー学会大会講演要旨集  17th-  2015
• Anti-inflammatory effect of a novel phospholipid, phosphatidyl-panthenol

  細川 雅史  コスメトロジー研究報告  23-  111  -116  2015  [Not refereed][Not invited]
  
• ニュートリゲノミクス解析に基づく褐藻カロテノイド,フコキサンチンの栄養機能性 (特集 ニュートリゲノミクス : 学問分野としての成熟とこれから) -- (現在のニュートリゲノミクス研究)

  宮下 和夫, 細川 雅史  遺伝 : 生物の科学  69-  (1)  21  -27  2015/01  [Not refereed][Not invited]
  
• 海洋細菌が有するカロテノイド合成系の体系的理解:フラボバクテリアにおけるmyxol合成系の特徴

  中西誠人, 高谷直己, 高峰, 宮下和夫, 細川雅史, 大熊盛也, 大島健志朗, 服部正平, 澤辺智雄  日本水産学会大会講演要旨集  2014-  34  2014/09/19  [Not refereed][Not invited]
  
• PPARγを活性化させる新規ニシン発酵食品の脂質解析

  濱岡直裕, 中川良二, 太田智樹, 長島浩二, 細川雅史, 宮下和夫  日本油化学会年会講演要旨集  53rd-  86  2014/09/09  [Not refereed][Not invited]
  
• Preventive Effects of Marine Carotenoids on Inflammation-related Diseases

  細川 雅史  微量栄養素研究  31-  80  -87  2014  [Not refereed][Not invited]
  
• Promotion of UCP1 Expression by Brown Seaweed Carotenoid, Fucoxanthin

  宮下 和夫, 細川 雅史  Bio industry  30-  (11)  31  -38  2013/11  [Not refereed][Not invited]
  
• 第二回日韓水産学会合同シンポジュウムに参加して

  細川 雅史  日本水産學會誌 = Bulletin of the Japanese Society of Scientific Fisheries  79-  (4)  744  -745  2013/07/15  [Not refereed][Not invited]
  
• 海洋細菌Jejuia pallidilutea A1株の新規カロテノイド合成とストレス応答

  高谷直己, 西田健太郎, 澤辺智雄, 宮下和夫, 細川雅史  マリンバイオテクノロジー学会大会講演要旨集  15th-  2013
• Carotenoid Contents in Chlorella pyrenoidosa and Concentration by Saponification

  加茂川 寛之, 細川 雅史, 阿部 真幸, 宮下 和夫  北海道大学水産科学研究彙報  62-  (3)  83  -88  2012/12/14  [Not refereed][Not invited]
   

  Carotenoids are natural pigments found in plants, seaweeds, algae and bacteria. It is well know that carotenoids have physiological effects such as provitamin activity and antioxidative effect. Among carotenoids, -carotene, lutein and zeaxanthin are popular carotenoids in green leafy vegetables and green microalgae. Furthermore, violaxanthin and neoxanthin are also contained as minor carotenoids. These carotenoids are currently expected to exhibit health beneficial effects depend on their unique structure. However, there are only a few reports of quantitative analysis of neoxanthin and vio...
• 脂肪を燃やす二つの脂肪組織(バイオミディア)

  細川 雅史  生物工学会誌 : seibutsu-kogaku kaishi  90-  (11)  740  -740  2012/11/25  [Not refereed][Not invited]
  
• 脂肪を燃やす二つの脂肪組織

  細川雅史  生物工学会誌  90-  (11)  740  2012/11/25  [Not refereed][Not invited]
  
• Molecular mechanism of anti-obesity effect by fucoxanthin

  HOSOKAWA MASASHI  日本水産學會誌 = Bulletin of the Japanese Society of Scientific Fisheries  78-  (5)  1007  -1007  2012/09/15  [Not refereed][Not invited]
  
• ヘマトコッカス藻由来アスタキサンチンの潰瘍性大腸炎予防効果

  木澤有希, 山下栄次, 宮下和夫, 細川雅史  日本水産学会大会講演要旨集  2012-  74  2012/09/14  [Not refereed][Not invited]
  
• 高度不飽和脂肪酸含有糖脂質の酸化安定性に関する研究

  山口智代, 細川雅史, 阿部真幸, 宮下和夫  日本水産学会大会講演要旨集  2012-  111  2012/03/26  [Not refereed][Not invited]
  
• Functionalities of Brown Seaweed Lipids and its Application to Foods

  宮下 和夫, 細川 雅史  Bio industry  29-  (2)  7  -14  2012/02  [Not refereed][Not invited]
  
• I-5. Molecular mechanism of anti-obesity effect by fucoxanthin

  Masashi Hosokawa  Nippon Suisan Gakkaishi (Japanese Edition)  78-  (5)  1007  -1007  2012  [Not refereed][Not invited]
  
• Study on phosphatidylation of functional compounds using phospholipases

  山本 幸弘, 細川 雅史, 宮下 和夫  成蹊大学理工学研究報告  48-  (2)  59  -62  2011/12  [Not refereed][Not invited]
   

  Here I describe about the phosphatidylation of functional compounds using phospholipases which is covered with my doctoral thesis. Physiologically active compound such as conjugated fatty acid, terpenes and phenolic compounds sometime exert their activity not fully because of their poor solubility in a certain environment. On the other hand, phospholipids have characteristic amphiphilic property and biocompatibility. In this study, several physiologically active compounds were phosphatidylated using phospholipases such as phospholipase A2 and phospholipase D. The enzymatic reaction was modified and optimized for the yield. In addition, the phosphatidylated compounds were tested for their physiological activity in vitro. Conjugated linoleic acids which have anti-obesity effects were phosphatidylated mediated by phospholipase A2. In this reaction, it is fund that water mimics such as formamide was key factor for the synthetic reaction. Several terpenes and phenolic compounds which have anti-cancer effects or anti-oxidative effects were phosphatidylated mediated by phospholipase D. Some phosphatidylated compounds were exerted higher anti-cancer effects compare to parent compounds.
• 褐藻類に含まれる機能性n‐3系PUFAの酸化安定性

  須田正剛, 阿部真幸, 細川雅史, 宮下和夫  日本水産学会大会講演要旨集  2011-  104  2011/09/28  [Not refereed][Not invited]
  
• 褐藻脂質中のフコキサンチン含有と脂肪酸組成の季節変動

  植西洋平, 野村雅俊, 阿部真幸, 細川雅史, 宮下和夫, 安井肇  日本水産学会大会講演要旨集  2011-  104  2011/09/28  [Not refereed][Not invited]
  
• 渇藻カロテノイド,フコキサンチンの生体内抗酸化活性

  AIRANTHI Widjaja‐Adhi, 岩崎さやか, 山口智代, 阿部真幸, 阿部真幸, 細川雅史, 宮下和夫  日本食品科学工学会大会講演集  58th-  91  2011/09/09  [Not refereed][Not invited]
  
• Anti-obesity and anti-diabetic effects by marinechemical "fucoxanthin"

  細川 雅史  Materials integration  24-  (8)  6  -10  2011/08  [Not refereed][Not invited]
  
• Preface

  Ernesto M. Hernandez, Masashi Hosokawa  Omega-3 Oils: Applications in Functional Foods  v  2011/05/01  [Not refereed][Not invited]
  
• Omega-3 Oils: Applications in Functional Foods

  Ernesto M. Hernandez, Masashi Hosokawa  Omega-3 Oils: Applications in Functional Foods  1  -305  2011/05/01  [Not refereed][Not invited]
   

  This book addresses new applications of omega-3 fatty acids from both plant and marine sources in food supplements and pharmaceuticals and covers three basic areas: structure and function, production and processing, and health effects. The authors review the latest clinical evidence on the impact of consumption of omega-3 polyunsaturated fatty acids on prevalent human diseases such as inflammation-related illnesses in general and cardiovascular illnesses in particular. They also examine technologies to purify marine oils and protect them against oxidation as well as novel techniques for their incorporation into foods.
• Synergistic/Additive Health Effects of Fish Oil and Bio-Active Compounds

  Tomoko Okada, Nana Mikami, Tomoyuki Okumura, Masashi Hosokawa, Kazuo Miyashita  Omega-3 Oils: Applications in Functional Foods  177  -203  2011/05/01  [Not refereed][Not invited]
   

  This chapter describes the current body of knowledge regarding the additive effects of fish oil and bio-active agents on the prevention and remediation of clinical disorders. It discusses the singular effect of fish oil consumption in relation to a specific illness, followed by the reported synergistic/additive effects of fish oil combined with bio-active agents. There is substantial evidence that fish oil possesses various physiological activity benefits in human health. The chapter discusses concomitant applications of fish oil with bio-active compounds for the remediation of a variety of clinical disorders. By combinative therapy, the well-documented beneficial health effect of fish oil can possibly be maximized in various approaches. Although this area of research is still in its early stages, it has been receiving increasing attention with the realization that combinative application of fish oil and bio-active compounds may provide vital outcomes and facilitate progress toward the design of beneficial clinical therapies for human health. Studies aimed at finding superior combinative effects of fish oil and bio-active compounds, along with their mechanisms of action, may eventually lead to reductions in the mortality and morbidity rates associated with human health disorders and complications that are of growing concern in modern society.
• 2-I-29 フコキサンチンと活性型ビタミンD_3による複合的大腸がん細胞増殖抑制の作用解析(一般演題要旨,日本ビタミン学会第63回大会講演要旨)

  寺崎 将, 長尾 昭彦, 田中 卓二, 細川 雅史, 宮下 和夫, 増田 園子  ビタミン  85-  (4)  250  -250  2011/04/25
• ホタテ卵巣を用いた新規発酵食品の製造と脂質の酸化安定性に関する検討

  濱岡直裕, 濱岡直裕, 太田智樹, 細川雅史, 宮下和夫  日本食品科学工学会大会講演集  58th-  2011
• カロテノイド(8)フコキサンチン

  細川 雅史  Functional food  5-  (1)  94  -98  2011  [Not refereed][Not invited]
  
• 2-I-16 活性型ビタミンD_3,Fucoxanthin,Shikoninによる複合的大腸がん細胞増殖抑制作用(一般演題,日本ビタミン学会第62回大会発表要旨)

  寺崎 将, 高上馬 希重, 関崎 春雄, 長尾 明彦, 田中 卓二, 細川 雅史, 宮下 和夫, 増田 園子  ビタミン  84-  (4)  174  -174  2010/04/25
• アロニアエタノール抽出物の脂質代謝改善効果

  梨田幸恵, 三上奈々, 細川雅史, 太田智樹, 宮下和夫  日本食品科学工学会大会講演集  57th-  2010
• Future direction of lipid nutrition and development of functional foods: gift from seaweeds and its application

  宮下 和夫, 細川 雅史  Journal of lipid nutrition  19-  (1)  39  -45  2010  [Not refereed][Not invited]
  
• アスタキサンチンのデキストラン硫酸ナトリウム誘発マウス大腸炎に対する予防効果

  細川 雅史, 安井 由美子, 三瓶 雄司, 三上 奈々, 宮下 和夫, 田中 卓二  日本栄養・食糧学会大会講演要旨集  63回-  192  -192  2009/05  [Not refereed][Not invited]
  
• 海洋生命統御による食糧生産--北海道大学21世紀COEプログラム(4)新たな水産資源としての海藻の重要性

  宮下 和夫, 細川 雅史  アクアネット  11-  (12)  78  -82  2008/12  [Not refereed][Not invited]
  
• AGFD 62-The suppression effects of conjugated linolenic acids on chemically induced colon carcinogenesis.

  Yumiko Yasui, Masashi Hosokawa, Kazuo Miyashita, Takuji Tanaka  ABSTRACTS OF PAPERS OF THE AMERICAN CHEMICAL SOCIETY  236-  2008/08  [Not refereed][Not invited]
  
• 内臓脂肪蓄積と食品機能成分:特にフコキサンチンについて (肥満と機能性食品)

  前多 隼人, 細川 雅史, 宮下 和夫  Functional food  1-  (2)  159  -165  2008  [Not refereed][Not invited]
  
• Chemopreventive effects of conjugated linolenic acids (CLN) occurring in plant seed oils

  Bhaskar Narayan, Masashi Hosokawa, Yumiko Yasui, Rikako Suzuki, Hiroyuki Kohno, Takuji Tanaka, Kazuo Miyashita  Current Nutrition and Food Science  3-  (4)  309  -318  2007  [Not refereed][Not invited]
   

  Conjugated fatty acids (CFA) being novel type of biologically functional lipid substances, have received increased interest owing to their beneficial effects on human health. Among CFAs, conjugated linoleic acids (CLA) have been reviewed extensively for their occurrence, health benefits, industrial production and applications. On the contrary, it is worth noting that some plant seeds contain conjugated linolenic acids (CLN) at considerably higher levels (30 to 70 wt% lipid), although other kinds of CFAs including CLA are only found at concentrations less than 1% in natural products. It is thus imperative that CLN are the only CFAs that occur in higher quantities in nature and we have extensively screened CLN from different plant seed oils for their chemopreventive effects. In the ensuing review, we describe in detail the physiological functions of CLN isomers that occur in some plant seeds. CLN from natural origin showed growth inhibitory and apoptotic effects on cancer cells and the activity was dependent on their geometrical/positional structures. All trans CLN exerted stronger growth inhibition and more DNA fragmentation in human colon cancer cells than corresponding CLN isomer with cis configuration. CLN induced apoptosis through decrease of Bcl-2 protein. It will be interacted with increase in PPARγ signaling and up-regulation of gene expression of GADD45 and p53. Enhancement of PPARγ expression was also observed in the rats fed seed oil rich in CLN. Dietary feeding of these CLN-containing seed oil reduced the frequency of colonic aberrant crypt foci (ACF) in rat that is the precursor of colon carcinogenesis in short-term experiment and inhibited the development of colonic adenocarcinoma in long-term experiment. © 2007 Bentham Science Publishers Ltd.
• Study on Biofunctional Properties and Increased Utilization of Marine Lipids

  HOSOKAWA Masashi  Oleoscience  7-  (12)  525  -532  2007  [Not refereed][Not invited]
   

  Marine organisms contain carotenoids with unique structure, and highly unsaturated fatty acids (HUFA) with health beneficial effects. Fucoxanthin, from the edible seaweed <I>Undaria pinnatifida</I>, induced apoptosis in human leukemia and colon cancer cells. Furthermore, the activities of apoptosis induction by halocynthiaxanthin and fucoxanthinol isolated from the sea squirt <I>Halocynthia roretzi</I> as metabolites of fucoxanthin were higher than that of fucoxanthin. In addition, dietary fucoxanthin attenuated the weight gain of white adipose tissue and decreased blood glucose in obese/diabetic mice. These results show multi-functional properties of fucoxanthin and its metabolites. On the other hand, to extend the application of HUFAs, enzymatic preparation of phospholipid (PL) binding HUFA and bioconversion of HUFA to hydroxyl cyclic fatty acid have been attempted. Phospholipase A<SUB>2</SUB> and Lipozyme IM show specificities on <I>sn</I>-2 and <I>sn</I>-1 position of PL, respectively, in esterification and acidolysis. Utilization of water mimics was effective in increasing the reaction yield of HUFA-PL. Further, tetrahydrofuranyl hydroxyl and diepoxy bicyclic fatty acids were converted from HUFA by <I>Bacillus megateriurn</I> ALA2.
• Study on biofunctional properties and increased utilization of marine lipids

  細川 雅史  Oleoscience  7-  (12)  525  -532  2007  [Not refereed][Not invited]
  
• Fucoxanthin, from edible seaweeds, and its multi-biological functions

  Kazuo Miyashita, Masashi Hosokawa, Tokutake Sashima  ABSTRACTS OF PAPERS OF THE AMERICAN CHEMICAL SOCIETY  232-  279  -279  2006/09  [Not refereed][Not invited]
  
• Physiological effects of eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) - A review

  B Narayan, K Miyashita, M Hosokawa  FOOD REVIEWS INTERNATIONAL  22-  (3)  291  -307  2006/07  [Not refereed][Not invited]
   

  Marine lipids have long been documented to be the major source of polyunsaturated fatty acids (PUFAs), especially n-3 fatty acids such as eicosapentaenoic acid (EPA; 20:5 n-3) and docosahexaenoic acid (DHA; 22:6 n-3). Both EPA and DHA have been documented to have significant influence on biochemical and physiological changes in the body. Although these long chain PUFA exert positive influences on human nutrition and health, there are also some controversies pertaining to the functioning of these n-3 PUFAs including the extent of their requirement by the body. As marine lipids have been thoroughly reviewed often, the present review mainly focuses on works related to physiological effects of EPA and DHA.
• 北海道支部の活性化 : 支部大会での新たな試み(支部のページ)

  細川 雅史, 澤谷 拓治, 山崎 浩司, 武田 浩郁, 天野 春菜  日本水産学会誌  71-  (3)  385  -387  2005/05/15  [Not refereed][Not invited]
  
• 海藻由来フコキサンチンの脂肪細胞に対する分化調節機能と抗肥満効果 (特集:フィトケミカルの機能性(2))

  前多 隼人, 細川 雅史, 宮下 和夫  食品工業  48-  (8)  68  -74  2005/04/30  [Not refereed][Not invited]
  
• 海藻脂質の知られざる健康機能 : 褐藻フコキサンチンに脂肪蓄積抑制などの機能

  細川 雅史, 宮下 和夫  化学と生物  43-  (3)  150  -152  2005/03/01  [Not refereed][Not invited]
  
• 「がん予防食品開発の新展開 －予防医学におけるバイオマーカーの評価システム－」<大澤俊彦編>

  シーエムシー出版  2005  [Not refereed][Not invited]
  
• Yasui, Y., Hosokawa, M., Kohno, H., Tanaka, T. and Miyashita, K.:" Chemopreventive effect of bitter gpourd seed oil rich in conjugated linolenic acid", Chapter 3, 30-41 (Hung, Y.S., Yanagita, T. and Knapp, H.R., AOCS Press, Champain, IL, USA)(2005)

  2005  [Not refereed][Not invited]
  
• Hosokawa, M. and Takahashi, K.:"Preparation of polyunsaturated phospholipids and their functional properties", Chapter 13, 13.1-13.13 (Hou, C.T.:"Handbook of Industrial Biocatalysis", CRC Press Taylor & Francis, Boca Raton, FL, USA)(2005)

  2005  [Not refereed][Not invited]
  
• 「油脂・脂質の基礎と応用－栄養・健康から工業まで－」<日本油化学会編>

  日本油化学会  2005  [Not refereed][Not invited]
  
• Takahashi, K. and Hosokawa, M.:"Functionalities and production with biocatalysis of two highly polyunsaturated phospholipids", Chapter 19, 274-284(Hung, Y.S., Yanagita, T. and Knapp, H.R., AOCS Press, Champain, IL, USA)(2005)

  2005  [Not refereed][Not invited]
  
• BIO R&D がん細胞を自滅させる海藻カロテノイド--フコキサンチンによるアポトーシス誘導

  細川 雅史  バイオインダストリー  21-  (1)  52  -57  2004/01  [Not refereed][Not invited]
  
• 油化学辞典

  丸善  2004  [Not refereed][Not invited]
  
• 『水産食品栄養学-基礎からヒトへ-』<鈴木平光編>

  技報堂  2004  [Not refereed][Not invited]
  
• ファインケミカルシリーズ『機能性脂質のフロンティア』<佐藤清隆・柳田晃良・和田俊 編>

  シーエムシー出版  2004  [Not refereed][Not invited]
  
• 水産学シリーズ142『水産機能性脂質』<高橋是太郎>

  恒星社厚生閣  2004  [Not refereed][Not invited]
  
• Proteolytic conversion of late run chum salmon muscle to angiotensin I-converting enzyme inhibitory peptides with TermoaseR

  Ono Seigo, Hosokawa Masashi, Miyasita Kazuo, Takahashi Koretarao  Bulletin of Fisheries Sciences, Hokkaido University  54-  (1)  1  -5  2003/07  [Not refereed][Not invited]
  
• Fujimoto, A., Sassaki, J., Hosokawa, M. and Takahashi, K.:"Fibrosarcoma may be suppressed by liposomal sn-2 DHA inserted phospolipid therapy", In Essential Fatty Acids and Eicosanoids Invited Paper from the Fifth International Congress, AOCS Press, Cha・・・

  2003  [Not refereed][Not invited]
   

  Fujimoto, A., Sassaki, J., Hosokawa, M. and Takahashi, K.:"Fibrosarcoma may be suppressed by liposomal sn-2 DHA inserted phospolipid therapy", In Essential Fatty Acids and Eicosanoids Invited Paper from the Fifth International Congress, AOCS Press, Champain (Y.S. Hung, S.J. Lin, P.C. Hung ed), (2003)pp. 290-295.
• Hou, C.T., Hosokawa, M. and Iwasaki, Y.:"Bioconversion of unsaturated fatty acids to value-added functional products", In Essential Fatty Acids and Eicosanoids Invited Paper from the Fifth International Congress, AOCS Press, Champain (Y.S. Hung, S.J. L・・・

  2003  [Not refereed][Not invited]
   

  Hou, C.T., Hosokawa, M. and Iwasaki, Y.:"Bioconversion of unsaturated fatty acids to value-added functional products", In Essential Fatty Acids and Eicosanoids Invited Paper from the Fifth International Congress, AOCS Press, Champain (Y.S. Hung, S.J. Lin, P.C. Hung ed), (2003)pp. 12-17.
• Hosokawa, M., Hou, C.T., Weisleder, D. and Brown, W."Formation of cyclic products from polyunsaturated fatty acids by Clavibacter sp. ALA2", In Essential Fatty Acids and Eicosanoids Invited Paper from the Fifth International Congress, AOCS Press, Champ・・・

  2003  [Not refereed][Not invited]
   

  Hosokawa, M., Hou, C.T., Weisleder, D. and Brown, W."Formation of cyclic products from polyunsaturated fatty acids by Clavibacter sp. ALA2", In Essential Fatty Acids and Eicosanoids Invited Paper from the Fifth International Congress, AOCS Press, Champain (Y.S. Hung, S.J. Lin, P.C. Hung ed), (2003)pp. 36-41.
• Noguchi, R., Yasui, Y., Hosokawa, M., Fukunaga, K. and Miyashita, K.:"Bioconversion of CLN to CLA", In Essential Fatty Acids and Eicosanoids Invited Paper from the Fifth International Congress, AOCS Press, Champain (Y.S. Hung, S.J. Lin, P.C. Hung ed), ・・・

  2003  [Not refereed][Not invited]
   

  Noguchi, R., Yasui, Y., Hosokawa, M., Fukunaga, K. and Miyashita, K.:"Bioconversion of CLN to CLA", In Essential Fatty Acids and Eicosanoids Invited Paper from the Fifth International Congress, AOCS Press, Champain (Y.S. Hung, S.J. Lin, P.C. Hung ed), (2003)pp. 353-359.
• Preparation of highly unsaturated phospholipid molecular species and functional properties

  M Hosokawa  NIPPON SUISAN GAKKAISHI  68-  (4)  509  -512  2002/07  [Not refereed][Not invited]
  
• Molecular Structuring of Highly Polyunsaturated Phospholipids and Their Biological Functionalities

  HOSOKAWA MASASHI  Bulletin of the Japanese Society of Scientific Fisheries  68-  (3)  297  -297  2002/05/15  [Not refereed][Not invited]
  
• Bioconversion of Highly Unsaturated Phospholipids and It's Biological Functions

  HOSOKAWA Masashi, TAKAHASHI Koretaro  Oleoscience : Journal of Japan Oil Chemists' Society  2-  (1)  19  -25  2002/01/01  [Not refereed][Not invited]
   

  Biocatalytic approaches were rational ways to prepare and modify highly unsaturated phospholipids (HUFA-PL). The prepared HUFA-PL showed an antitumor effect in vivo, a promotional effect on cell differentiation, and an increase in deformability of erythrocytes <I>in Vitro</I>.
• 高度不飽和リン脂質リポソームの性状とその抗腫瘍効果

  藤本 章人, 佐々木 甚一, 細川 雅史, 高橋 是太郎  脂質生化学研究  43-  (0)  318  -321  2001/05/10  [Not refereed][Not invited]
   

  高度不飽和リン脂質は多様な機能を有する生体適合性(生体親和性)に優れた脂質形態である。本研究では高度不飽和リン脂質リポソームの免疫賦活促進機能を調べることを目的として、フィトグリコーゲン(PG)を内包させた高度不飽和リン脂質リポソーム及びPG未内包高度不飽和リン脂質リポソームを調製し、それらの性状と in vitro, in vivo 相方における抗腫瘍効果について検討した。調製したsn-2位DHA結合型リン脂質リポソーム(PC:PS=7:3 モル比)は6～9層からなるマルチラメラベシクル(MLV)であり、粒度分布のピークは約1 μ m付近にあった。PGを内包させなかったsn-2位DHA結合型リン脂質MLVは少なくとも1ヶ月以上の粒度分布安定性を示し、Meth-A fibrosarcomaに対して比較的強い抗腫瘍効果を示した。
• 第4版油化学便覧<油化学便覧編集委員会>

  丸善  2001  [Not refereed][Not invited]
  
• Takahashi, K. and Hosokawa, M.: “Chapter 25 Production of tailor-made polyunsaturated phospholipids through bioconversions”, 517-526 (Kuo, T.M., Gardner, H.W.: "Lipid Biotechnology", MARCEL DEKKER, INC., New York) (2001)

  2001  [Not refereed][Not invited]
  
• DHA含有リン脂質の酵素変換と生理作用 (「リン脂質の生理作用」)

  高橋 是太郎, 細川 雅史  脂質栄養学  9-  (1)  19  -31  2000  [Not refereed][Not invited]
  
• β-リゾ型高度不飽和リン脂質の調製及びヒト赤血球変形能改善効果

  小野 雅代, 細川 雅史, 高橋 是太郎, 井上 良計  脂質生化学研究  40-  (0)  204  -206  1998/07/02  [Not refereed][Not invited]
   

  Rhizomucor miehei 由来の固定化リパーゼ剤（リポザイムIM）の水分活性を中間水分活性域に制御することによってDHA強化鶏卵黄リン脂質を部分加水分解し、DHA含量が極めて高いβ-リゾ型リン脂質が得られた。このβ-リゾ型リン脂質は低濃度で速やかに赤血球に取り込まれ、赤血球の変形能を改善した。
• Beneficial Effect of β-lysophospholipid Rich in Highly Unsaturated Fatty Acids on Deformability of Human Erythrocyte

  小野 雅代, 細川 雅史, 高橋 是太郎, 井上 良計  ヘモレオロジー研究会誌  1-  (0)  7  -11  1998  [Not refereed][Not invited]
   

  高度不飽和リン脂質には多彩な生理活性のあることが示唆され始めている。高度不飽和リン脂質は水産物に比較的豊富に含まれてはいるが，水産物より着臭や着色なしに抽出することは一般に難しい。一方，ドコサヘキサエン酸(DHA)を強化した鶏卵黄リン脂質は，着臭や着色が殆どないという利点を有しているが，水産物由来のリン脂質よりもDHA含量が低いという難点があった。我々はこのような点を解決するため，リパーゼによる鶏卵黄リン脂質の部分加水分解反応を試み，DHAを高濃度に含むリン脂質の調製法を確立した。本研究では，このように調製したβーリゾ型高度不飽和リン脂質の生理機能を検討するため，ヒト赤血球変形能改善効果について人工毛細血管モデル(MC-FAN)を用いて評価した。Immobilized lipase (Lipozyme IM) mediated selective partial hydrolysis of docosahexaenoic acid (DHA) enriched egg yolk phospholipids (PL) were carried out. By keeping the water activity of lipase at an intermediate level， DHA was effectively enriched in acyl moieties of PL. This DHA-PL was dominant in β-lysophospholipids，and has rapidly improved the deformability of human erythrocytes when evaluated with artificial capillary model.β- lysophospholipids might be one of the most effective chemical forms to incorporate DHA into cells，accordingly.
• Concentration of Docosahexaenoic Acid-Contain-ing Phospholipid through Lipozyme IM-mediated Hydrolysis(共著)

  ONO Masayo, HOSOKAWA Masashi, INOUE Yoshikazu, TAKAHASHI Koretaro  Journal of Japan Oil Chemists' Society  46-  (8)  867  -872,936  1997  [Not refereed][Not invited]
   

  A selective partial hydrolyses of egg-yolk lecithin from fish-oil fed hens and squid-skin lecithin were carried out with immobilized Rhizomucor miehei lipase (Lipozyme IM) in n-hexane media in order to concentrate docosahexaenoic acid (DHA) -containing phospholipids. DHA were abundant in phospholipid fractions that had recovered from the partial hydrolysates than the corresponding substrates i.e. egg-yolk lecithin and squid lecithin. Thus, it was clear that DHA ester bonds were less susceptible against hydrolysis than other fatty acid ester bonds. Optimum water activity (a_w) for Lipozyme IM to concentrate DHA-containing phospholipids was a_w=0.44. Under this a_w, DHA showed to three fold increase compared to that of the original egg-yolk lecithin from fish-oil fed hens, after 8 h reaction at 40°C. Water activity-controlled partial hydrolysis with Lipozyme IM was borned out to be a useful method for concentrating functional phospholipids.

Industrial Property Rights

• 血糖値上昇抑制剤

  特開２００７－２９７３７０
• フコキサンチノールの製造方法

  特願2005-340997
• 生体内ＤＨＡ合成促進剤

  特開２００７－７７０６７
• 抗腫瘍剤

  特願2004-060821.

Awards & Honors

• 2014 アスタキサンチン研究会 9th Astaxanthin Award
   

  受賞者: HOSOKAWA Masashi
• 2012 日本油化学会 WCOS2012 Outstanding Poster Presentation Award
   

  受賞者: 細川 雅史
• 2007 平成18年度日本油化学会進歩賞
  
• 2003 The American Oil Chemists' Society, Biotechnology Division, Outstanding Paper Presentation.
  
• 2003 The American Oil Chemists' Society, Health & Nutrition Devision, Best Overall Poster Award.
  
• 2002 平成14年度 日本油化学会オレオサイエンス賞
  
• 2001 平成13年度 日本水産学会奨励賞
  

Research Grants & Projects

• Functional regulation of immune cells in the metabolic disease prevention mechanism by brown seaweed fucoxanthin

  Japan Society for the Promotion of Science：Grants-in-Aid for Scientific Research

  Date (from‐to) : 2021/04 -2024/03 

  Author : 細川 雅史

   

  2021年度の研究において、以下の研究成果を得た。 (1)フコキサンチンを投与した肥満マウスの脂肪組織では、これまで報告しているように活性化マクロファージマーカーのF4/80およびM1マクロファージマーカーのCD11cのmRNA発現量の低下が確認された。さらに、M1マーカー(CD11c)/M2マーカー(Arg1)比も低下したことから、フコキサンチンがマクロファージの脂肪組織への浸潤を抑制するとともに、極性変化にも関わることが推察された。一方、T細胞マーカーを解析した結果、CD8に加えCD25およびCD4のmRNA発現量が低下し、T細胞サブセットへの影響が示唆された。(2)非アルコール性脂肪肝炎を誘導したマウスの肝臓では、フコキサンチンの投与によりCD8およびCD25のmRNA発現量が低下傾向を示した。(3)肝細胞およびマクロファージ株をフコキサンチン代謝物のフコキサンチノールで処理し、各オルガネラにおける蓄積をHPLCにて定量した。細胞膜、ミトコンドリア、ミクロソーム画分でフコキサンチノールが検出されたことから、細胞膜を通過しオルガネラへの移行することが推察された。特に、ミトコンドリア画分での蓄積が多いことが示唆された。(4)フコキサンチノールがミトコンドリア画分に蓄積していた点に着目し、ミトコンドリア機能に関わるミトコンドリアDNA量の測定を行った。NASHモデルマウスの肝臓からDNAを抽出し測定を行った結果、普通食群と比較してNASH誘導色群でみられたミトコンドリアDNA量の低下が、フコキサンチンにより抑制された。(5)エネルギー代謝に関わる白色脂肪組織から褐色脂肪組織への転換メカニズムとして分泌因子を介した細胞間コミュニケーションについて調べた。その結果、BMPを介した組織転換制御の可能性が認められた。
• Enzymatic conversion of kelp biomass for biofuels and health beneficial products.

  Japan Society for the Promotion of Science：Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (B)

  Date (from‐to) : 2020/04 -2024/03 

  Author : FOX BRIAN, 高須賀 太一, 細川 雅史, 栗原 秀幸

   

  Our aim of the current project is to develop an efficient method to depolymerize natural kelps available in the Hokkaido ocean by a combination of several enzyme functions, including laminarin-degrading enzymes and alginate-degrading enzymes. A set of kelp spp. were obtained and ready to analytically evaluate their chemical compositions for the next fiscal year. Regarding a laminarin-degrading enzyme, we had picked up a glycoside hydrolase 55 enzyme (GH55) from a highly cellulolytic insect symbiont bacterium (Bianchetti and Takasuka et al., 2015, Journal of Biological Chemistry). 7 polysaccharide lyases 18 enzymes (PL18s), which function in alginate degradation, were selected from 7 different bacterial species, and four PL18s had been recombinantly expressed and subjected to the initial enzyme screening by using pure alginic acid substrate. Moreover, two polysaccharide lyase 17 enzymes (PL17s), were gene synthesized and cloned into the bacterial expression vector. From the previous studies, PL18 is shown to be the endo-type polysaccharide lyase, which produces oligoalginic acids (>3 degrees of polymerization (DP)), while PL17 should catalyze an exo-type reaction. In the FY2021, collected kelps will be tested to be hydrolyzed by at least three different enzymes, GH55, PL17, and PL18, and end products will be analyzed.
• Developmental research of marine carotenoids on prevention of lifestyle-related diseases through their metabolite analysis and regulation of organ/cell interaction

  Japan Society for the Promotion of Science：Grants-in-Aid for Scientific Research

  Date (from‐to) : 2018/04 -2021/03 

  Author : Hosokawa Masashi

   

  A fucoxanthin-derived apocarotenoid, paracentrone was identified in liver and white adipose tissue of mice fed fucoxanthin. Further, dietary fucoxanthin attenuated hepatic fat accumulation and inflammation in the diet-induced nonalcoholic steatohepatitis model mice. Paracentrone suppressed the overexpression of inflammatory factors in activated RAW264.7 macrophages. Further, apo-10′-fucoxanthinal with an epoxide residue also suppressed pro-inflammatory cytokines through regulating IkB phosphorylation, while beta-apo-8′-carotenal did not suppress inflammation. In adipocytes and hepatocytes, paracentrone and apo-10′-fucoxanthinal downregulated the mRNA expression of proinflammatory mediators induced by the culture medium of activated RAW264.7 cells. These results suggest that apo-fucoxanthinoids derived from fucoxanthin regulate organ/cell interaction involved in chronic inflammation, which is basis of the development of lifestyle-related diseases.
• Establishment of cold stress-resistance nori strains specialized for the mariculture in Tohoku in Japan

  Japan Society for the Promotion of Science：Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (B)

  Date (from‐to) : 2015/04 -2018/03 

  Author : Mikami Koji

   

  Tohoku area is the northern limit of the large scale-nori mariculture and has had a severe problem such as reduction of nori productivity in the early spring because of lowering temperature of the sea water, which affects negatively on seaweed growth. The present study therefore focuses on the establishment of a new strain enabling the sustainable production of nori in Tohoku area. To establish cold stress-resistance strains, a strain whose growth tolerates to cold stress was first selected with the help from the Aichi Fisheries Research Institute. Next, the crossing of this strain with Kesen-nuna Susabi, generally used as a cultivation strain in Tohoku area, was performed and then several lines exhibiting characteristics different from both parental strains were finally isolated. Thus, it is necessary to confirm growth under cold stress conditions of these new strains, which could enable to establish novel strains that are adapting to mariculture in Tohoku area in near future.
• Preventive effects on metabolic syndrome through adipose tissue browning induced by marine carotenoids

  Japan Society for the Promotion of Science：Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (B)

  Date (from‐to) : 2015/04 -2018/03 

  Author : HOSOKAWA Masashi, KATSUKI Ami, OHUCHI Yuka, TYA Daisuke, TATEYAMA Riho, AKITA Tomoki

   

  Fucoxanthin contained in brown seaweeds suppressed weight gain of white adipose tissue (WAT) through induction of mitochondrion factors including uncoupling protein 1 (UCP1) in adipose tissues. In diet-induce obese mice fed fucoxanthin, fatty acid oxidation was promoted. Induction of the brown adipocyte-like phenotype was also observed in C3H10T1/2 adipocytes treated with fucoxanthin metabolites. PGC-1alpha is suggested to be a critical factor to induce browning in white adipocytes by fucoxanthin. On the other hand, the number of progenitor cells to differentiate into beige adipocytes was demonstrated to decrease with aging.
• Creation of A Light-Driven Marine Vibrio Factory for Biofuel Production

  Japan Society for the Promotion of Science：Grants-in-Aid for Scientific Research Grant-in-Aid for Challenging Exploratory Research

  Date (from‐to) : 2014/04 -2016/03 

  Author : Sawabe Tomoo, HOSOKAWA MASASHI

   

  An aim of the study is to develop light-driven marine vibrio biocatalysts in efficient biofuel production. We performed 1) gene expression of a Nonlabens sediminis proteorhdopsin gene, and 2) a cloning of a myxol synthetic gene cluster of Nonlabens ulvanivorans. We also accumulated fundamental knowledge on CRISPR/Cas system for further developments of a genome editing approach working in marine vibrio cells.
• Development of high perfomanced hydogen production system using marine vibrios as biocatalysts

  Japan Society for the Promotion of Science：Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (B)

  Date (from‐to) : 2013/04 -2016/03 

  Author : Sawabe Tomoo, HOSOKAWA MASASHI, NAKAGAWA SATOSHI

   

  Aims of the study are to establishment of 1) continuous culture system and 2) biorefinery of hydrogen producing marine vibrios as biocatalysts. We did succeeded 1) continuous hydrogen production for 70 days with appropriate hydrogen producing rates and yields, 2) alginate-hydrogen conversion using 2 kinds of biocatalysts, and 3) bioprospects of new and rare carotenoid producers and their genome information.
• Biological Localization and Functionality of Allenic Carotenoids

  Japan Society for the Promotion of Science：Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (B)

  Date (from‐to) : 2011/04 -2014/03 

  Author : MIYASHITA Kazuo, HOSOKAWA Masashi

   

  In the present study, molecular mechanisms were made clear for the anti-obesity effect of fucoxanthin by the induction of UCP1 in abdominal WAT. The anti-obesity effect of fucoxanthin was suggested to be due to its specific structure that contains two carotenoid end rings of the polyene chromophore containing allenic bond and OH and epoxy groups. Therefore, other allenic carotenoids or scission products from fucoxanthin could not show such activities as found in fucoxanthin. Fucoxanthin also improves insulin resistance and decreases blood glucose level through the regulation of adipocytokine secretions from the abdominal WAT. Another important molecular mechanism for the improvement of glucose utilization by fucoxanthin will be glucose transporter 4 (GLUT4) up-regulation and promotion of its translocation to cell membrane from cytosol.
• Preventive effects of marine carotenoids on chronic inflammation-related diseases through regulation of cell and tissue communication

  Japan Society for the Promotion of Science：Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (B)

  Date (from‐to) : 2011/04 -2014/03 

  Author : HOSOKAWA Masashi, YASUI Yumiko

   

  Fucoxanthin (Fx) suppressed chronic inflammation in white adipose tissue by the inhibition of macrophage and T cell infiltration. Further, Fx down-regulated mRNA expression of chemokines and adipokines, which recruit immune cells and induce insulin resistance. In diabetic/obese mice fed Fx, GLUT4 activation was observed in skeletal muscle through the improvement of hyperglycemia. On the other hand, Fx and astaxanthin (Asx) extracted from H. pluvialis prevent DSS-induced colitis and colon carcinogenesis in mice. In colon of the mice fed Asx, mRNA expression levels of inflammatory cytokines tended to decrease compared to control mice. In addition, Asx down-regulated mRNA expression of inflammatory cytokines in macrophage-like cells and colon cells stimulated by the culture medium of activated macrophages. These results suggest that marine carotenoids prevent chronic inflammation and its-related diseases by the regulation of cell and tissue communication via chemokines and cytokines.
• Creation of marine vibrio cells with minimun gene sets for efficient biofuel production

  Japan Society for the Promotion of Science：Grants-in-Aid for Scientific Research Grant-in-Aid for Challenging Exploratory Research

  Date (from‐to) : 2011 -2013 

  Author : SAWABE Tomoo, HOSOKAWA Masashi, NAKAGAWA Satoshi

   

  In maintaining a sustainable ecosystem in this period of global warming, development of key technologies in the field of renewable energy sources has become an important challenge; a method of biofuel production from marine biomass could be one of the most crucial technologies in the future. In this study, we performed global transcriptome, fermentation product profiling, and metabolic engineering to design marine vibrio cells with efficient ethanol production from marine biomass.
• Regulation of carotenoid biosynthesis in non- photosynthetic bacteria by light irradiation

  Japan Society for the Promotion of Science：Grants-in-Aid for Scientific Research Grant-in-Aid for Challenging Exploratory Research

  Date (from‐to) : 2011 -2012 

  Author : HOSOKAWA Masashi

   

  11ShimoA1, which was isolated from a marine alga, synthesized a novel carotenoid,2’-isopentenyl saporxanthin (2’-isopentenyl Spx). Blue LED irradiation increased2’-isopentenyl Spx content in the culture medium of strain A1. Furthermore, blue LED also enhanced mRNA expression level of carotenoid synthetic genes such as crtB、crtI、crtY、crtZ. In addition, strain A1 has unique genes coded prenyltransferase and phytoene desaturase, which are suggested to play an important role for 2’-isopentenyl Spx synthesis in strain A1. Both genes also tended to be up-regulated by blue LED. These results show that strain A1 is a unique bacteria with carotenoid synthetic pathways regulated by blue light.
• Preventive effects of marine carotenoids on life-style-related diseases based on anti-inflammatory effects as molecular targets

  Japan Society for the Promotion of Science：Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (B)

  Date (from‐to) : 2008 -2010 

  Author : HOSOKAWA Masashi, YASUI Yumiko

   

  Astaxanthin exhibited inhibitory effects on dextran sodium sulfate-induced colitis and inflammatory-related colon carcinogenesis in mice. In colon of the mice fed astaxanthin, mRNA expression levels of inflammatory cytokines such as TNF-α and IL-1β were suppressed through inhibition of NF-κB activation. After feeding for 20 weeks, colonic mucosal ulcers, dysplastic crypts and colonic adenocarcinoma were inhibited by 200 ppm astaxanthin-containing diet. In addition, astaxanthin, pectenolone, alloxanthin and diatoxanthin isolated from marine organisms attenuated excessive mRNA expression of inflammatory cytokines and COX-2 and iNOS in macrophages activated by LPS. On the other hand, dietary 0.2% fucoxanthin markedly suppressed mRNA expression of inflammatory cytokines such as TNF-α and IL-6 to induce insulin resistance and inhibited macrophage infiltration in white adipose tissue through suppression of MCP-1 mRNA expression after 4 weeks feeding. Further, fucoxanthinol, which is one of fucoxanthin metabolites, also regulated cross talk of adipocytes and macrophages via palmitic acid and TNF-α. These results suggest that anti-inflammatory effects are molecular mechanism for anti-obesity and anti-diabetic effects of fucoxanthin.
• Regulation of adipocytes differentiation by marine lipids

  Japan Society for the Promotion of Science：Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (C)

  Date (from‐to) : 2005 -2007 

  Author : HOSOKAWA Masashi

   

  Obesity is now recognized as a world wide problem. It is multiple risk factors for the life style-related diseases such as diabetes, hypertension, hyperlipidemia, and cardiovascular disease. Visceral fat accumulation plays crucial roles in the development of these diseases. So, it is important to prevent the life style-related diseases through significantly reducing excess accumulation of visceral fat. Fucoxanthin is a major carotenoid found in edible seaweed such as Undaria pinnatifida and Hijikia fusiformis. Fucoxanthin and its metabolite, fucoxanthinol, inhibited intercellular lipid accumulation and suppressed glycerol-3-phosphate dehydrogenase activity during adipocyte differentiation of 3T3-L1 cells. In addition, in 3T3-L1 cells treated with fucoxanthin and fucoxanthinol, peroxisome proliferators-activated receptor (PPAR_γ), which regulates adipogenic gene expression, was down-regulated in a dose-dependent manner. N-3 Highly unsaturated fatty acid, docosahexaenoic acid (DHA) and eicosapentaenoic acid (EPA), which are major fatty acids in fish oil, also suppressed adipocyte differentiation of 3T3-L1 cells by down regulation PPAR_γ And C/EBP_α. White adipose tissue weight of mice fed fucoxanthin was significantly lower than that of mice fed control diet. Uncoupling protein 1(UCP1), a key molecule for metabolic thermogenesis, was clearly found in WAT of mice fed fucoxanthin. The anti-obesity effect of fucoxanthin was enhanced by the combination with fish oil containing DHA and EPA
• Development of Oxidative Stability and Bioavailability of Fatty Nutraceuticals Using Single Dispersed Micro/Nanosphere

  Japan Society for the Promotion of Science：Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (B)

  Date (from‐to) : 2004 -2006 

  Author : MIYASHITA Kazuo, HOSOKAWA Masashi

   

  1.The oxidative stability of EPA and DHA in aqueous solutions was dramatically different from that in bulk phase. When the oxidative stabilities of six types of typical PUFA were compared in micelles, LA was the most susceptible to oxidation. In contrast, DHA and EPA were very stable. Oxidative stability of EPA and DHA in emulsion depended on the particle size and the interface. With decreasing particle size the stability of DHA and EPA increased. The reverse result was obtained in the oxidation of linoleate. 2.The oxidative stability of lipids in emulsion is strongly affected by altering the droplet size and the packing of the emulsifier and lipid molecules at the interface. Oxidative stability was compared among TGs dispersed with three kinds of sucrose esters having the same fatty acyl composition, the stability increased with increasing their hydrophile-lipophile balance (HLB) in both TGs, although these emulsifiers had no effect on the oxidative stability of SoyTG and DHATG in bulk phase. 3.Antioxidant activity also depended on the interface. With increasing the concentration of antioxidatns in interface layer the stability of lipids increased. 4.Micro/nanosphere containing functional lipid compounds can be stabilized by the regulation of interface. Moreover, it would be more acceptable to biological systems than other forms.
• 核内転写因子の発現を制御する高度不飽和リン脂質に関する研究

  日本学術振興会：科学研究費助成事業 若手研究(B)

  Date (from‐to) : 2002 -2003 

  Author : 細川 雅史

   

  昨年度の研究において、DHA含有ホスファチジルエタノールアミン(DHA-PE)がdibutyrylcAMP(dbcAMP)によるヒト白血病HL-60細胞の分化誘導を顕著に亢進することを明らかにした。更にその過程で核内転写因子のAP-1を構成するc-Junタンパク質の発現増加を確認した。AP-1は、c-Jun/c-Junのホモダーマーまたはc-Jun/c-Fos、c-Jun/ATF-2のヘテロダイマーによって構成され、MAPキナーゼにより活性化されることで分化に関連する遺伝子を発現する。これらの結果を踏まえ、本年度は以下の点を明らかにした。 1.DHA-PEとdbcAMPを併用した場合c-Fosタンパク質並びにそのmRNAの強い発現がみられたが、dbcAMPの単独処理におけるc-Fos発現量との間に大きな差が見られなかった。よって、dbcAMPによる分化の誘導にはc-Fosおよびc-Junの双方が重要な役割を果しているが、DHA-PEとの併用効果に発現にはc-Junタンパクがキーレギュレーターとなっていることが示唆された。 2.DHA-PEとdbcAMPにより併用処理したHL-60細胞では、MAPキナーゼの一つであるJNKが培養初期において一過的に活性化された。これによってAP-1の転写活性も高められ、分化に関連する遺伝子の発現が亢進されることが示唆された。 3.DHAをリガンドとすることが知られている核内受容体のペルオキシソーム増殖薬応答性受容体(PPAR)γの発現への影響を調べたが、その発現への影響は認められなかった。 以上の結果から、DHA-PEは分化誘導物質であるdbcAMPと併用することにより、c-Junの発現亢進とJNKの活性化により核内転写因子AP-1の転写活性を高め、ヒト白血病HL-60細胞への分化誘導効果を亢進することが示唆された。
• DHA結合型リン脂質の構造と白血病細胞の分化誘導機構に関する研究

  日本学術振興会：科学研究費助成事業 奨励研究(A)

  Date (from‐to) : 1997 -1998 

  Author : 細川 雅史

   

  1. DHA結合型リン脂質の分化誘導促進効果:(1)シロサケ筋肉,精巣及びマイカ肝膵臓から抽出した水産リン脂質が,ヒト白血病(HL-60)細胞に対するレチノイン酸やジブチリルcAMPなどの分化誘導剤の作用を増強する効果,すなわち分化誘導促進能を有することが明らかとなった。(2)マウス白血病(M1)細胞に対しては,DHA結合型リン脂質単独においても分化誘導能が認められた。(3)分化誘導促進効果は,DHAの遊離脂肪酸形態に比べリン脂質形態,特にPE形態において顕著なものであったことから,機能発現にはリン脂質形態が重要であると推察された。 2. 細胞膜への影響:TNBSクエンチング法並びに脂肪酸組成分析におけるDHA含有率の上昇から,DHA結合型リン脂質がHL-60細胞内に取り込まれていることを確認した。 3. 機能遺伝子発現への影響:DHA結合型PEは,レチノイン酸の分化誘導過程において重要な役割を果たすレチノイドXレセプタ-α(RXRα)のmRNA量を増加させることが示唆された。 4. 細胞内情報伝達酵素への影響:DHA結合型リン脂質の分化誘導促進作用がプロテインキナーゼ阻害剤により抑制されたことから,分化誘導促進能はプロテインキナーゼを介した作用であることが示唆された。 以上の結果から,水産リン脂質を含めたDHA結合型リン脂質が白血病細胞に対して分化誘導能及び分化誘導促進能を有することが明らかとなった。機能発現にはリン脂質の構造が重要であり,細胞に取り込まれた後,細胞内情報伝達酵素や核レセプターの発現を介して分化誘導剤の作用を促進することが推察された。
• ガン細胞の増殖、分化に及ぼすDNA含有リン脂質の影響に関する研究

  日本学術振興会：科学研究費助成事業 奨励研究(A)

  Date (from‐to) : 1995 -1995 

  Author : 細川 雅史

   

  酸素反応により種々のドコサヘキサエン酸(DHA)含有リン脂質を調製し、ヒト前骨髄性白血病細胞(HL-60細胞)の増殖、分化に及ぼす影響について検討した。 1.種々のDHA含有ホスファチジルコリン(PC)分子種をHL-60細胞培養液に添加して、あらかじめ24時間培養することにより、レチノイン酸(RA)の細胞分化誘導能が増強された(細胞分化誘導促進効果)。その効果はDNA含有PC分子種により異なり、1-Pal,2-DHA-PC及び1-Ole,2-DHA-PCにおいて顕著であった。 2.DHA含有PCの細胞分化誘導促進効果は濃度依存的に上昇し、それに伴い細胞内のDHA含量の増加が見られた。DHAの結合していないdi-Pal-PC、di-Ole-PC及び大豆PCには促進効果が見られなかった。 3.DHA含有リン脂質の化学形態の影響について検討したところ、同一分子種ではPCに比べ、ポスファチジルエタノールアミン(PE)、ホスファチジルセリン(PS)の形態において細胞分化誘導能の増強作用が大きく、特に1-Pal,2-DHA-PE及び1-Ole,2-DHA-PEにおいて細胞分化誘導促進効果が顕著であった。更に、DHA含有PE自体にも分化誘導作用が見られた。 4.HL-60細胞の増殖に及ぼすDHA含有リン脂質分子種の影響についても検討を行ったところ、DHA含有PEの形態において増殖抑制効果が大きかった。 以上の結果より、DHA含有リン脂質がガン細胞の分化誘導促進能や増殖抑制能を有し、その効果がリン脂質の分子種や化学形態によって異なることが明らかとなった。特に、1-Pal,2-DHA-PE及び1-Ole,2-DHA-PEの効果が顕著であったことから、制ガン剤の減量とともに副作用の軽減への利用が期待される。
• 赤血球の変形能に及ぼす高度不飽和脂肪酸含有リン脂質の影響に関する研究

  日本学術振興会：科学研究費助成事業 奨励研究(A)

  Date (from‐to) : 1994 -1994 

  Author : 細川 雅史

   

  1.赤血球の変形能をシリコン単結晶基板マイクロチャンネルを用いて測定することにより、再現性の高い測定結果が得られた。 2.リパーゼやホスホリパーゼを用いて合成したEPA,DHA含有ホスファチジルコリン(PC)は、赤血球にとり込まれることによりその変形能を向上させることを証明した。 3.エタノールアミン結合リン脂質ではsn-2位にDHAを含むものが、セリン結合リン脂質ではsn-1位にEPAを含むものが赤血球の変態能を大きく向上させることも証明できた。 4.リン脂質クラス間を比較すると、エタノールアミン結合リン脂質はコリン結合リン脂質およびセリン結合リン脂質よりも、赤血球変形能を向上させる作用が強いことを認めた。特に、sn-2位にDHAを含むエタノールアミン結合リン脂質が赤血球の変形能を最も向上できることが示唆された。 5.蛍光偏光解消法により赤血球膜の流動性を測定したところ、膜の流動性向上は赤血球の変形能の向上とよく対応していた。 6.鮭卵より調製したEPA,DHA含有リン脂質においても赤血球変形能の向上作用がみられた。 7.リノール酸を主な脂肪酸とする大豆リン脂質は赤血球の変形能を改善しなかった。また、水素添加したリン脂質は赤血球の変形能を明かに低下させた。 以上の結果により、EPA,DHA含有リン脂質が赤血球の変形能を向上させ、その作用の強弱が分子種や脂質形態によって異なることが明かとなった。血栓症などの成人病予防が大きな課題となっている今日、EPA,DHA含有リン脂質を合成し、機能の一部を分子種レベルで明かにできたことは意義のあることと考える。
• 赤血球膜流動性におよぼす高度不飽和脂肪酸含有複合脂質の影響に関する研究

  日本学術振興会：科学研究費助成事業 奨励研究(A)

  Date (from‐to) : 1993 -1993 

  Author : 細川 雅史

   

  1.ブタ膵臓ホスホリパーゼA_2によるsn-2位にEPA,DHAを含むホスファチジルコリンの合成、LipozymeIM60によるsn-1位にEPAを含むホスファチジルコリンの合成、Streptomyces sp.由来のホスホリパーゼDによるこれらのホスファチジルコリンのホスファチジル基転移を行うことができきた。これによりホスファチジルコリンと同一の脂肪酸残基をもつ高度不飽和脂肪酸含有ホスファチジルエタノールアミンとホスファチジルセリンの合成物を得ることができた。 2.コリン結合リン脂質に比べアミノ基結合リン脂質のほうが赤血球により導入されやすいことを明らかにできた。 3.TNBSクエンチング法により、赤血球にとり込まれたリン脂質の40〜50%は膜タンパク質近傍に存在することを示すことができた。 4.高度不飽和脂肪酸含有ホスファチジルエタノールアミンはほとんど溶血性を示さないことがわかった。 5.水素添加したリン脂質は赤血球膜の流動性を明らかに低下させることを確認できた。 6.大豆リン脂質は赤血球膜の流動性を改善できないことがわかった。 7.合成したEPA,DHA含有リン脂質はいずれも赤血球膜流動性を改善する作用のあることが確認された。 8.コリン結合リン脂質、アミノ基結合リン脂質はいずれもEPA,DHAをsn-2位に含む混合形態の場合に最も高い赤血球膜流動性改善効果を示すことがわかった。 9.同一の脂肪酸残基の場合、コリン結合リン脂質はアミノ基結合リン脂質よりも赤血球膜の流動性改善効果が低いことをみとめた。 以上の結果より判断し、高度不飽和脂肪酸含有リン脂質は赤血球膜の流動性を高めて赤血球の変形能を向上させ、ひいては高血圧症や血栓症の予防や治療に役立つことも期待できることが明かとなった。
• Enzymatic synthesis of phospholipid containing polyunsaturated fatty acid and its pysiological functionality

  Japan Society for the Promotion of Science：Grants-in-Aid for Scientific Research Grant-in-Aid for General Scientific Research (C)

  Date (from‐to) : 1992 -1993 

  Author : TAKAHASHI Koretaro, HOSOKAWA Masashi, HATANO Mutsuo

   

  Synthesis of phospholipid containing polyunsaturated fatty acid were carried out. 1. Incorporation of polyunsaturated fatty acid into position sn^<-2> of phospholipid was possible by the use of an industrial pancreatic phospholipase A_2 mediated esterification. using lysophosphatidylcholine isolated from a commercial emulsifier and mixed fatty acids from sardine oil or high purity EPA.DHA. Incorporation of polyunsaturated fatty acid into position sn^<-1> of phospholipid was also possible by the use of an industrial immobilized lipase e.g. Lipozyme mediated transesterification. using phosphatidylcholine isolated from soy lecithin and mixed fatty acids from sardine oil or high purity EPA.DHA. Over the medium range water activity of the synthetic reactor promoted the incorporation of polyunsaturated fatty acid into phospholipid. but at the same time. unfavorable hydrolysis has also been proceeded. and as a result, it has decreased the recovery of phospholipid. Under the low water activity condition of the reactor, the incorporation of polyunsaturated fatty acid into phospholipid was sluggish. However, the recovery of phospholipid was relatively high. Precised control of water activity and the amount of essential water substitutes e.g. polyalcohol made it possible to achieve maximum incorporation rate of polyunsaturated fatty acid into phospholipid, and at the same time, made it possible to obtain high recovery of phospholipid. The rate of retinoic acid induced differentiation of the human promyelocytic lcukemia cell line (HL-60) was promoted by growing HL-60 cells in FBN-PNM pretreated with cither EPA containing phosphatidylcholine or DHA containing phosphatidylcholine.
• Highly utilization of marine biofunctional materials

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• 2022年4月1日 

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  2024年3月31日 

  大学院水産科学研究院副研究院長