Please use this identifier to cite or link to this item: http://hdl.handle.net/11455/93031
標題: Study on the Antileukemic Immunity of Dunalella salina and Hepatoprotective Activity of Hydrogen-Rich Water in Mice
探討杜莎藻對血癌小鼠免疫調節作用及富氫水對小鼠護肝作用
作者: Wen-Chen Chuang
莊文貞
關鍵字: 杜莎藻;抗血癌;免疫反應;富氫水;水飛薊素;四氯化碳;抗發炎;小鼠;Dunaliella salina;anti-leukemia;immune responses;Hydrogen-rich water;Silymarin;CCl4;anti-oxidation;anti-inflammation;mice
引用: part 1 1. Aharon O (2005) A hundred years of Dunaliella research: 1905–2005. Saline Syst 1: 1-14. 2. Avron M (1986) The osmotic components of halotolerant alga. Trends Biochem Sci 11: 5-6. 3. Lers A, Biener Y, Zamir A (1990) Photoinduction of Massive beta-Carotene Accumulation by the Alga Dunaliella bardawil: Kinetics and Dependence on Gene Activation. Plant Physiol 93: 389-395. 4. Ginzburg M (1987) Dunaliella: a green alga adapted to salt. Adv Bot Res 14: 93-183. 5. Tsugane S, Tsuda M, F Gey aSW (1992) Cross-sectional study with multiple measurements of biological markers for assessing stomach cancer risks at the population level. Environ Health Perspect 98: 207-210. 6. Massyuk N (1966) Mass culture of the carotene containing alga Dunaliella salina Teod. Ukr Bot Zh 23: 12-19. 7. Hosseini Tafreshi A, Shariati M (2009) Dunaliella biotechnology: methods and applications. J Appl Microbiol 107: 14-35. 8. Ben-Amotz A, Katz A, Avron M (1982) Accumulation of β-carotene in halotolerant algae: purification and characterization of β-carotene-rich globules from Dunaliella bardawil (Chlorophyceae). J Phycol 18: 529-537. 9. Garcia-Gonzalez M, Moreno J, Manzano JC, Florencio FJ, Guerrero MG (2005) Production of Dunaliella salina biomass rich in 9-cis-beta-carotene and lutein in a closed tubular photobioreactor. J Biotechnol 115: 81-90. 10. Hu CC, Lin JT, Lu FJ, Chou FP, Yang D-J (2008) Determination of carotenoids in Dunaliella salina cultivated in Taiwan and antioxidant capacity of the algal carotenoid extract. Food Chemistry 109: 439-446. 11. Klausner A (1986) Algaculture: food for thought. Bio Technology 4: 947-953. 12. Olson JA (1989) Provitamin A function of carotenoids: the conversion of beta-carotene into vitamin A. J Nutr 119: 105-108. 13. Levin G, Yeshurun M, Mokady S (1997) In vivo antiperoxidative effect of 9-cis beta-carotene compared with that of the all-trans isomer. Nutr Cancer 27: 293-297. 14. Cooper DA, Eldridge AL, Peters JC (1999) Dietary carotenoids and certain cancers, heart disease, and age-related macular degeneration: a review of recent research. Nutr Rev 57: 201-214. 15. Li Y, Ma A, Shao X, Du Z (2008) [Study the effect of antioxidant vitamin E, vitamin C and beta-carotene supplement on erythrocyte functions in elderly person]. Wei Sheng Yan Jiu 37: 305-308. 16. Palozza P, Serini S, Torsello A, Boninsegna A, Covacci V, et al. (2002) Regulation of cell cycle progression and apoptosis by beta-carotene in undifferentiated and differentiated HL-60 leukemia cells: possible involvement of a redox mechanism. Int J Cancer 97: 593-600. 17. Palozza P, Serini S, Maggiano N, Angelini M, Boninsegna A, et al. (2002) Induction of cell cycle arrest and apoptosis in human colon adenocarcinoma cell lines by beta-carotene through down-regulation of cyclin A and Bcl-2 family proteins. Carcinogenesis 23: 11-18. 18. Palozza P, Serini S, Torsello A, Di Nicuolo F, Maggiano N, et al. (2003) Mechanism of activation of caspase cascade during beta-carotene-induced apoptosis in human tumor cells. Nutr Cancer 47: 76-87. 19. Lowe GM, Booth LA, Young AJ, Bilton RF (1999) Lycopene and beta-carotene protect against oxidative damage in HT29 cells at low concentrations but rapidly lose this capacity at higher doses. Free Radic Res 30: 141-151. 20. Zhuang S, Lynch MC, Kochevar IE (1999) Caspase-8 mediates caspase-3 activation and cytochrome c release during singlet oxygen-induced apoptosis of HL-60 cells. Exp Cell Res 250: 203-212. 21. Chlichlia K, Peter ME, Rocha M, Scaffidi C, Bucur M, et al. (1998) Caspase activation is required for nitric oxide-mediated, CD95(APO-1/Fas)-dependent and independent apoptosis in human neoplastic lymphoid cells. Blood 91: 4311-4320. 22. Bendich A (1989) Carotenoids and the immune response. J Nutr 119: 112-115. 23. Bendich A (1991) Beta-carotene and the immune response. Proc Nutr Soc 50: 263-274. 24. Michal JJ, Heirman LR, Wong TS, Chew BP, Frigg M, et al. (1994) Modulatory effects of dietary beta-carotene on blood and mammary leukocyte function in periparturient dairy cows. J Dairy Sci 77: 1408-1421. 25. Chew BP, Park JS, Weng BC, Wong TS, Hayek MG, et al. (2000) Dietary beta-carotene is taken up by blood plasma and leukocytes in dogs. J Nutr 130: 1788-1791. 26. Kuroiwa Y, Nishikawa A, Imazawa T, Kitamura Y, Kanki K, et al. (2006) A subchronic toxicity study of dunaliella carotene in F344 rats. Food Chem Toxicol 44: 138-145. 27. Ben-Amotz A, Avron M (1983) Accumulation of metabolites by halotolerant algae and its industrial potential. Annu Rev Microbiol 37: 95-119. 28. Levin G, Mokady S (1994) Antioxidant activity of 9-cis compared to all-trans beta-carotene in vitro. Free Radic Biol Med 17: 77-82. 29. Ben-Amotz A, Levy Y (1996) Bioavailability of a natural isomer mixture compared with synthetic all-trans beta-carotene in human serum. Am J Clin Nutr 63: 729-734. 30. Ben-Amotz A, Rachmilevich B, Greenberg S, Sela M, Weshler Z (1996) Natural beta-carotene and whole body irradiation in rats. Radiat Environ Biophys 35: 285-288. 31. Lavy A, Naveh Y, Coleman R, Mokady S, Werman MJ (2003) Dietary Dunaliella bardawil, a beta-carotene-rich alga, protects against acetic acid-induced small bowel inflammation in rats. Inflamm Bowel Dis 9: 372-379. 32. Chidambara Murthy KN, Vanitha A, Rajesha J, Mahadeva Swamy M, Sowmya PR, et al. (2005) In vivo antioxidant activity of carotenoids from Dunaliella salina--a green microalga. Life Sci 76: 1381-1390. 33. Raja R, Hemaiswarya S, Balasubramanyam D, Rengasamy R (2007) Protective effect of Dunaliella salina (Volvocales, Chlorophyta) against experimentally induced fibrosarcoma on wistar rats. Microbiol Res 162: 177-184. 34. Hsu YW, Tsai CF, Chang WH, Ho YC, Chen WK, et al. (2008) Protective effects of Dunaliella salina--a carotenoids-rich alga, against carbon tetrachloride-induced hepatotoxicity in mice. Food Chem Toxicol 46: 3311-3317. 35. Fujitani N, Sakaki S, Yamaguchi Y, Takenaka H (2001) Inhibitory effects of microalgae on the activation of hyaluronidase. J Appl Phycol 13: 489-492. 36. Tsai CF, Lu FJ, Hsu YW (2012) Protective effects of Dunaliella salina - a carotenoids-rich alga - against ultraviolet B-induced corneal oxidative damage in mice. Mol Vis 18: 1540-1547. 37. Ivanov EP, Tolochko GV, Shuvaeva LP, Ivanov VE, Iaroshevich RF, et al. (1998) Infant leukemia in Belarus after the Chernobyl accident. Radiat Environ Biophys 37: 53-55. 38. Scandura JM, Boccuni P, Cammenga J, Nimer SD (2002) Transcription factor fusions in acute leukemia: variations on a theme. Oncogene 21: 3422-3444. 39. Wang ZY (2002) Mechanism of action of all-trans retinoic acid and arsenic trioxide in the treatment of acute promyelocytic leukemia. Gan to Kagaku Ryoho 29 Suppl 1: 214-218. 40. Zhang X, Zhou H, Song S, Qiao Z, Yang L, et al. (2001) [The impact of arsenic trioxide or all-trans retinoic acid treatment on coagulopathy in acute promyelocytic leukemia]. Zhonghua Nei Ke Za Zhi 40: 829-833. 41. Muto A, Kizaki M, Yamato K, Kawai Y, Kamata-Matsushita M, et al. (1999) 1,25-Dihydroxyvitamin D3 induces differentiation of a retinoic acid-resistant acute promyelocytic leukemia cell line (UF-1) associated with expression of p21(WAF1/CIP1) and p27(KIP1). Blood 93: 2225-2233. 42. Liu YF, Shen ZX, Chen X, Wang AH, Cao Q, et al. (2003) [Clinical observation of the short-term efficacy of the treatment with combination of all-trans retinoic acid (ATRA) and arsenic trioxide (As2O3) in newly diagnosed acute promyelocytic leukemia (APL)]. Zhonghua Xue Ye Xue Za Zhi 24: 25-27. 43. Kuo HC, Kuo WH, Lee YJ, Wang CJ, Tseng TH (2006) Enhancement of caffeic acid phenethyl ester on all-trans retinoic acid-induced differentiation in human leukemia HL-60 cells. Toxicol Appl Pharmacol 216: 80-88. 44. Chang YC, Hsu JD, Lin WL, Lee YJ, Wang CJ (2012) High incidence of acute promyelocytic leukemia specifically induced by N-nitroso-N-methylurea (NMU) in Sprague-Dawley rats. Arch Toxicol 86: 315-327. 45. Giver CR, Wong R, Moore DH, 2nd, Pallavicini MG (2001) Persistence of aneuploid immature/primitive hemopoietic sub-populations in mice 8 months after benzene exposure in vivo. Mutat Res 491: 127-138. 46. Amsellem S, Pflumio F, Bardinet D, Izac B, Charneau P, et al. (2003) Ex vivo expansion of human hematopoietic stem cells by direct delivery of the HOXB4 homeoprotein. Nat Med 9: 1423-1427. 47. Lin JJ, Hsu HY, Yang JS, Lu KW, Wu RS, et al. (2011) Molecular evidence of anti-leukemia activity of gypenosides on human myeloid leukemia HL-60 cells in vitro and in vivo using a HL-60 cells murine xenograft model. Phytomedicine 18: 1075-1085. 48. Mitchel RE, Jackson JS, McCann RA, Boreham DR (1999) The adaptive response modifies latency for radiation-induced myeloid leukemia in CBA/H mice. Radiat Res 152: 273-279. 49. Herzog RW, Arruda VR (2003) Update on gene therapy for hereditary hematological disorders. Expert Rev Cardiovasc Ther 1: 215-232. 50. He Q, Na X (2001) The effects and mechanisms of a novel 2-aminosteroid on murine WEHI-3B leukemia cells in vitro and in vivo. Leuk Res 25: 455-461. 51. Glass B, Uharek L, Zeis M, Loeffler H, Mueller-Ruchholtz W, et al. (1996) Graft-versus-leukaemia activity can be predicted by natural cytotoxicity against leukaemia cells. Br J Haematol 93: 412-420. 52. Lai T-Y, Lin J-J, Huang W-W, Kuo S-C, Wen Y-F, et al. (2012) Arsenic trioxide (As2O3) inhibits murine WEHI-3 leukemia in BALB/c mice in vivo. Environ Toxicol 27: 364-371. 53. Lin JP, Yang JS, Lu CC, Chiang JH, Wu CL, et al. (2009) Rutin inhibits the proliferation of murine leukemia WEHI-3 cells in vivo and promotes immune response in vivo. Leuk Res 33: 823-828. 54. Schoenborn JR, Wilson CB (2007) Regulation of interferon-gamma during innate and adaptive immune responses. Adv Immunol 96: 41-101. 55. Schroder K, Hertzog PJ, Ravasi T, Hume DA (2004) Interferon-gamma: an overview of signals, mechanisms and functions. J Leukoc Biol 75: 163-189. 56. Cerretti DP, McKereghan K, Larsen A, Cantrell MA, Anderson D, et al. (1986) Cloning, sequence, and expression of bovine interleukin 2. Proc Natl Acad Sci U S A 83: 3223-3227. 57. Mott HR, Driscoll PC, Boyd J, Cooke RM, Weir MP, et al. (1992) Secondary structure of human interleukin 2 from 3D heteronuclear NMR experiments. Biochemistry 31: 7741-7744. 58. Lapinski PE, King PD (2012) Regulation of Ras signal transduction during T cell development and activation. Am J Clin Exp Immunol 1: 147-153. 59. Mayer R (2008) In: WILEY-VCH Verlag GmbH & Co. KGaA W, editor. Immunology-From Cell Biology to Disease Chapter 6-8. 60. Saraiva M, O'Garra A (2010) The regulation of IL-10 production by immune cells. Nat Rev Immunol 10: 170-181. 61. Mosser DM, Zhang X (2008) Interleukin-10: new perspectives on an old cytokine. Immunol Rev 226: 205-218. 62. Braat H, Rottiers P, Hommes DW, Huyghebaert N, Remaut E, et al. (2006) A phase I trial with transgenic bacteria expressing interleukin-10 in Crohn's disease. Clin Gastroenterol Hepatol 4: 754-759. 63. Hatanaka H, Abe Y, Naruke M, Tokunaga T, Oshika Y, et al. (2001) Significant correlation between interleukin 10 expression and vascularization through angiopoietin/TIE2 networks in non-small cell lung cancer. Clin Cancer Res 7: 1287-1292. 64. Mocellin S, Marincola FM, Young HA (2005) Interleukin-10 and the immune response against cancer: a counterpoint. J Leukoc Biol 78: 1043-1051. 65. Chan SL, Mo FK, Wong CS, Chan CM, Leung LK, et al. (2012) A study of circulating interleukin 10 in prognostication of unresectable hepatocellular carcinoma. Cancer 118: 3984-3992. 66. De Vita F, Orditura M, Galizia G, Romano C, Roscigno A, et al. (2000) Serum interleukin-10 levels as a prognostic factor in advanced non-small cell lung cancer patients. Chest 117: 365-373. 67. Hu CC, Lin JT, Lu FJ, Chou FP, Yang DJ (2008) Determination of carotenoids in Dunaliella salina cultivated in Taiwan and antioxidant capacity of the algal carotenoid extract. Food Chem 109: 439-446. 68. Sacha T, Zawada M, Hartwich J, Lach Z, Polus A, et al. (2005) The effect of β-carotene and its derivatives on cytotoxicity, differentiation, proliferative potential and apoptosis on the three human acute leukemia cell lines: U-937, HL-60 and TF-1. BBA - Mol Basis Dis 1740: 206-214. 69. Upadhyaya KR, Radha KS, Madhyastha HK (2007) Cell cycle regulation and induction of apoptosis by beta-carotene in U937 and HL-60 leukemia cells. J Biochem Mol Biol 40: 1009-1015. 70. Hsu YW, Tsai CF, Chang WH, Ho YC, Chen WK, et al. (2008) Protective effects of Dunaliella salina--a carotenoids-rich alga, against carbon tetrachloride-induced hepatotoxicity in mice. Food Chem Toxicol 46: 3311-3317. 71. Raja R, Hemaiswarya S, Balasubramanyam D, Rengasamy R (2007) Protective effect of Dunaliella salina (Volvocales, Chlorophyta) against experimentally induced fibrosarcoma on wistar rats. Microbiol Res 162: 177-184. 72. Fujii Y, Sakamoto S, Ben-Amotz A, Nagasawa H (1993) Effects of beta-carotene-rich algae Dunaliella bardawil on the dynamic changes of normal and neoplastic mammary cells and general metabolism in mice. Anticancer Res 13: 389-393. 73. Yang D-J, Lin J-T, Chen Y-C, Liu S-C, Lu F-J, et al. (2013) Suppressive effect of carotenoid extract of Dunaliella salina alga on production of LPS-stimulated pro-inflammatory mediators in RAW264.7 cells via NF-κB and JNK inactivation. J Func Foods 5: 607-615. 74. Lin HW, Chen YC, Liu CW, Yang DJ, Chen SY, et al. (2014) Regulation of virus-induced inflammatory response by Dunaliella salina alga extract in macrophages. Food Chem Toxicol 71: 159-165. 75. Lai TY, Lin JJ, Huang WW, Kuo SC, Wen YF, et al. (2012) Arsenic trioxide (As(2)O(3)) inhibits murine WEHI-3 leukemia in BALB/c mice in vivo. Environ Toxicol 27: 364-371. 76. Funk RK, Maxwell TJ, Izumi M, Edwin D, Kreisel F, et al. (2008) Quantitative trait loci associated with susceptibility to therapy-related acute murine promyelocytic leukemia in hCG-PML/RARA transgenic mice. Blood 112: 1434-1442. 77. Jayappriyan KR, Rajkumar R, Venkatakrishnan V, Nagaraj S, Rengasamy R (2013) In vitro anticancer activity of natural β-carotene from Dunaliella salina EU5891199 in PC-3 cells. Biomed Prev Nutr 3: 99-105. 78. Bechelli J, Coppage M, Rosell K, Liesveld J (2011) Cytotoxicity of Algae Extracts on Normal and Malignant Cells. Leukemia Res Treat 2011: 1-7. 79. Sheu MJ, Huang GJ, Wu CH, Chen JS, Chang HY, et al. (2008) Ethanol extract of Dunaliella salina induces cell cycle arrest and apoptosis in A549 human non-small cell lung cancer cells. In Vivo 22: 369-378. 80. Grivennikov SI, Greten FR, Karin M (2010) Immunity, inflammation, and cancer. Cell 140: 883-899. 81. Mantovani A, Sica A (2010) Macrophages, innate immunity and cancer: balance, tolerance, and diversity. Curr Opin Immunol 22: 231-237. 82. Sica A, Allavena P, Mantovani A (2008) Cancer related inflammation: The macrophage connection. Cancer Lett 267: 204-215. 83. Malmberg KJ, Bryceson YT, Carlsten M, Andersson S, Bjorklund A, et al. (2008) NK cell-mediated targeting of human cancer and possibilities for new means of immunotherapy. Cancer Immunol Immunother 57: 1541-1552. 84. Smyth MJ, Crowe NY, Pellicci DG, Kyparissoudis K, Kelly JM, et al. (2002) Sequential production of interferon-gamma by NK1.1(+) T cells and natural killer cells is essential for the antimetastatic effect of alpha-galactosylceramide. Blood 99: 1259-1266. 85. Smyth MJ, Cretney E, Kelly JM, Westwood JA, Street SE, et al. (2005) Activation of NK cell cytotoxicity. Mol Immunol 42: 501-510. 86. Akao Y, Ebihara T, Masuda H, Saeki Y, Akazawa T, et al. (2009) Enhancement of antitumor natural killer cell activation by orally administered Spirulina extract in mice. Cancer Sci 100: 1494-1501. 87. Chew BP, Park JS (2004) Carotenoid action on the immune response. J Nutr 134: 257S-261S. 88. Jing Y, Dai J, Chalmers-Redman RM, Tatton WG, Waxman S (1999) Arsenic trioxide selectively induces acute promyelocytic leukemia cell apoptosis via a hydrogen peroxide-dependent pathway. Blood 94: 2102-2111. 89. Chung JG, Yang JS, Huang LJ, Lee FY, Teng CM, et al. (2007) Proteomic approach to studying the cytotoxicity of YC-1 on U937 leukemia cells and antileukemia activity in orthotopic model of leukemia mice. Proteomics 7: 3305-3317. 90. Jorge Olmos RGaVPR (2015) Apoptosis Comparison Effects Between Synthetic and Natural Β-Carotene from Dunaliella salina on MDA-MB-231Brest Cancer Cells. J Microb Biochem Technol 7: 051-056. 91. Chidambara Murthy KN, Vanitha A, Rajesha J, Mahadeva Swamy M, Sowmya PR, et al. (2005) In vivo antioxidant activity of carotenoids from Dunaliella salina--a green microalga. Life Sci 76: 1381-1390. 92. Hieber AD, King TJ, Morioka S, Fukushima LH, Franke AA, et al. (2000) Comparative effects of all-trans beta-carotene vs. 9-cis beta-carotene on carcinogen-induced neoplastic transformation and connexin 43 expression in murine 10T1/2 cells and on the differentiation of human keratinocytes. Nutr Cancer 37: 234-244. 93. Peto R, Doll R, Buckley JD, Sporn MB (1981) Can dietary beta-carotene materially reduce human cancer rates? Nature 290: 201-208. 94. Ziegler RG, Colavito EA, Hartge P, McAdams MJ, Schoenberg JB, et al. (1996) Importance of alpha-carotene, beta-carotene, and other phytochemicals in the etiology of lung cancer. J Natl Cancer Inst 88: 612-615. 95. Paolini M, Cantelli-Forti G, Perocco P, Pedulli GF, Abdel-Rahman SZ, et al. (1999) Co-carcinogenic effect of beta-carotene. Nature 398: 760-761. 96. Paolini M, Abdel-Rahman SZ, Sapone A, Pedulli GF, Perocco P, et al. (2003) Beta-carotene: a cancer chemopreventive agent or a co-carcinogen? Mut Res 543: 195-200. 97. Albanes D, Heinonen OP, Taylor PR, Virtamo J, Edwards BK, et al. (1996) Alpha-Tocopherol and beta-carotene supplements and lung cancer incidence in the alpha-tocopherol, beta-carotene cancer prevention study: effects of base-line characteristics and study compliance. J Natl Cancer Inst 88: 1560-1570. 98. Omenn GS, Goodman GE, Thornquist MD, Balmes J, Cullen MR, et al. (1996) Risk factors for lung cancer and for intervention effects in CARET, the Beta-Carotene and Retinol Efficacy Trial. J Natl Cancer Inst 88: 1550-1559. 99. Wang XD, Russell RM (1999) Procarcinogenic and anticarcinogenic effects of beta-carotene. Nutr Rev 57: 263-272. 100. Cooper DA, Eldridge AL, Peters JC (1999) Dietary carotenoids and lung cancer: a review of recent research. Nutr Rev 57: 133-145. 101. Kim Y, Lian F, Yeum KJ, Chongviriyaphan N, Choi SW, et al. (2007) The effects of combined antioxidant (beta-carotene, alpha-tocopherol and ascorbic acid) supplementation on antioxidant capacity, DNA single-strand breaks and levels of insulin-like growth factor-1/IGF-binding protein 3 in the ferret model of lung cancer. Int J Cancer 120: 1847-1854. 102. Offord EA, Gautier JC, Avanti O, Scaletta C, Runge F, et al. (2002) Photoprotective potential of lycopene, beta-carotene, vitamin E, vitamin C and carnosic acid in UVA-irradiated human skin fibroblasts. Free Radic Biol Med 32: 1293-1303. 103. Liu C, Russell RM, Wang XD (2004) Alpha-tocopherol and ascorbic acid decrease the production of beta-apo-carotenals and increase the formation of retinoids from beta-carotene in the lung tissues of cigarette smoke-exposed ferrets in vitro. J Nutr 134: 426-430. 104. Palozza P (2005) Can beta-carotene regulate cell growth by a redox mechanism? An answer from cultured cells. Biochim Biophys Acta 1740: 215-221. 105. Kim Y, Chongviriyaphan N, Liu C, Russell RM, Wang XD (2006) Combined antioxidant (beta-carotene, alpha-tocopherol and ascorbic acid) supplementation increases the levels of lung retinoic acid and inhibits the activation of mitogen-activated protein kinase in the ferret lung cancer model. Carcinogenesis 27: 1410-1419. part 2 1. Ohsawa I, Ishikawa M, Takahashi K, Watanabe M, Nishimaki K, et al. (2007) Hydrogen acts as a therapeutic antioxidant by selectively reducing cytotoxic oxygen radicals. Nat Med 13: 688-694. 2. Zheng XF, Sun XJ, Xia ZF (2011) Hydrogen Resuscitation, a New Cytoprotective Approach. Clin Exp Pharmacol Physiol 38: 155-163. 3. Fritsch J, Lenz O, Friedrich B (2013) Structure, function and biosynthesis of O2-tolerant hydrogenases. Nat Rev Micro 11: 106-114. 4. Dole M, Wilson FR, Fife WP (1975) Hyperbaric hydrogen therapy: a possible treatment for cancer. Science 190: 152-154. 5. Gharib B, Hanna S, Abdallahi OM, Lepidi H, Gardette B, et al. (2001) Anti-inflammatory properties of molecular hydrogen: investigation on parasite-induced liver inflammation. C R Acad Sci III 324: 719-724. 6. Quaglia MG, Bossu E, Donati E, Mazzanti G, Brandt A (1999) Determination of silymarine in the extract from the dried silybum marianum fruits by high performance liquid chromatography and capillary electrophoresis. J Pharm Biomed Anal 19: 435-442. 7. Ball KR, Kowdley KV (2005) A review of Silybum marianum (milk thistle) as a treatment for alcoholic liver disease. J Clin Gastroenterol 39: 520-528. 8. Letteron P, Labbe G, Degott C, Berson A, Fromenty B, et al. (1990) Mechanism for the protective effects of silymarin against carbon tetrachloride-induced lipid peroxidation and hepatotoxicity in mice. Evidence that silymarin acts both as an inhibitor of metabolic activation and as a chain-breaking antioxidant. Biochem Pharmacol 39: 2027-2034. 9. Saliou C, Rihn B, Cillard J, Okamoto T, Packer L (1998) Selective inhibition of NF-kappaB activation by the flavonoid hepatoprotector silymarin in HepG2. Evidence for different activating pathways. FEBS Lett 440: 8-12. 10. Boigk G, Stroedter L, Herbst H, Waldschmidt J, Riecken EO, et al. (1997) Silymarin retards collagen accumulation in early and advanced biliary fibrosis secondary to complete bile duct obliteration in rats. Hepatology 26: 643-649. 11. Mourelle M, Franco MT (1991) Erythrocyte defects precede the onset of CCl4-induced liver cirrhosis. Protection by silymarin. Life Sci 48: 1083-1090. 12. Hsu YW, Tsai CF, Chuang WC, Chen WK, Ho YC, et al. (2010) Protective effects of silica hydride against carbon tetrachloride-induced hepatotoxicity in mice. Food Chem Toxicol 48: 1644-1653. 13. Tsai CF, Hsu YW, Chen WK, Chang WH, Yen CC, et al. (2009) Hepatoprotective effect of electrolyzed reduced water against carbon tetrachloride-induced liver damage in mice. Food Chem Toxicol 47: 2031-2036. 14. Hsu YW, Tsai CF, Chen WK, Lu FJ (2009) Protective effects of seabuckthorn (Hippophae rhamnoides L.) seed oil against carbon tetrachloride-induced hepatotoxicity in mice. Food Chem Toxicol 47: 2281-2288. 15. Hsu YW, Tsai CF, Chang WH, Ho YC, Chen WK, et al. (2008) Protective effects of Dunaliella salina--a carotenoids-rich alga, against carbon tetrachloride-induced hepatotoxicity in mice. Food Chem Toxicol 46: 3311-3317. 16. Hall PM, Plummer JL, Ilsley AH, Ahern MJ, Cmielewski PL, et al. (1994) The pathology of liver injury induced by the chronic administration of alcohol and 'low-dose' carbon tetrachloride in Porton rats. J Gastroenterol Hepatol 9: 250-256. 17. Hemmings SJ, Pulga VB, Tran ST, Uwiera RR (2002) Differential inhibitory effects of carbon tetrachloride on the hepatic plasma membrane, mitochondrial and endoplasmic reticular calcium transport systems: implications to hepatotoxicity. Cell Biochem Funct 20: 47-59. 18. Castillo T, Koop DR, Kamimura S, Triadafilopoulos G, Tsukamoto H (1992) Role of cytochrome P-450 2E1 in ethanol-, carbon tetrachloride- and iron-dependent microsomal lipid peroxidation. Hepatology 16: 992-996. 19. McGregor D, Lang M (1996) Carbon tetrachloride: genetic effects and other modes of action. Mutat Res 366: 181-195. 20. Weber LW, Boll M, Stampfl A (2003) Hepatotoxicity and mechanism of action of haloalkanes: carbon tetrachloride as a toxicological model. Crit Rev Toxicol 33: 105-136. 21. Tirmenstein MA, Ge X, Elkins CR, Fariss MW (1999) Administration of the tris salt of alpha-tocopheryl hemisuccinate inactivates CYP2E1, enhances microsomal alpha-tocopherol levels and protects against carbon tetrachloride-induced hepatotoxicity. Free Radic Biol Med 26: 825-835. 22. Wu J, Danielsson A, Zern MA (1999) Toxicity of hepatotoxins: new insights into mechanisms and therapy. Expert Opin Investig Drugs 8: 585-607. 23. Basu S (2003) Carbon tetrachloride-induced lipid peroxidation: eicosanoid formation and their regulation by antioxidant nutrients. Toxicology 189: 113-127. 24. Sheweita SA, El-Gabar MA, Bastawy M (2001) Carbon tetrachloride changes the activity of cytochrome P450 system in the liver of male rats: role of antioxidants. Toxicology 169: 83-92. 25. Kuz'menko DI, Laptev BI (1999) [Reserve of serum lipids for peroxidation during oxidative stress in rats]. Vopr Med Khim 45: 47-52. 26. Valko M, Rhodes CJ, Moncol J, Izakovic M, Mazur M (2006) Free radicals, metals and antioxidants in oxidative stress-induced cancer. Chem Biol Interact 160: 1-40. 27. Benzie IF (2000) Evolution of antioxidant defence mechanisms. Eur J Nutr 39: 53-61. 28. Cross CE, van der Vliet A, O'Neill CA, Louie S, Halliwell B (1994) Oxidants, antioxidants, and respiratory tract lining fluids. Environ Health Perspect 102 Suppl 10: 185-191. 29. Nakane T, Asayama K, Kodera K, Hayashibe H, Uchida N, et al. (1998) Effect of selenium deficiency on cellular and extracellular glutathione peroxidases: immunochemical detection and mRNA analysis in rat kidney and serum. Free Radic Biol Med 25: 504-511. 30. Roxborough HE, Mercer C, McMaster D, Maxwell AP, Young IS (1999) Plasma glutathione peroxidase activity is reduced in haemodialysis patients. Nephron 81: 278-283. 31. Wunder C, Potter RF (2003) The heme oxygenase system: its role in liver inflammation. Curr Drug Targets Cardiovasc Haematol Disord 3: 199-208. 32. Wu G, Fang YZ, Yang S, Lupton JR, Turner ND (2004) Glutathione metabolism and its implications for health. J Nutr 134: 489-492. 33. Esterbauer H, Dieber-Rotheneder M, Striegl G, Waeg G (1991) Role of vitamin E in preventing the oxidation of low-density lipoprotein. Am J Clin Nutr 53: 314S-321S. 34. Hybertson BM, Gao B, Bose SK, McCord JM (2011) Oxidative stress in health and disease: the therapeutic potential of Nrf2 activation. Mol Aspects Med 32: 234-246. 35. Kyriakis JM, Avruch J (2012) Mammalian MAPK signal transduction pathways activated by stress and inflammation: a 10-year update. Physiol Rev 92: 689-737. 36. Lawrence T (2009) The nuclear factor NF-kappaB pathway in inflammation. Cold Spring Harb Perspect Biol 1: a001651. 37. Ke Q, Costa M (2006) Hypoxia-inducible factor-1 (HIF-1). Mol Pharmacol 70: 1469-1480. 38. van Uden P, Kenneth NS, Rocha S (2008) Regulation of hypoxia-inducible factor-1alpha by NF-kappaB. Biochem J 412: 477-484. 39. Fukuda K, Asoh S, Ishikawa M, Yamamoto Y, Ohsawa I, et al. (2007) Inhalation of hydrogen gas suppresses hepatic injury caused by ischemia/reperfusion through reducing oxidative stress. Biochem Biophys Res Commun 361: 670-674. 40. Liu Y, Yang L, Tao K, Vizcaychipi MP, Lloyd DM, et al. (2014) Protective effects of hydrogen enriched saline on liver ischemia reperfusion injury by reducing oxidative stress and HMGB1 release. BMC Gastroenterol 14: 12. 41. Xiang L, Tan JW, Huang LJ, Jia L, Liu YQ, et al. (2012) Inhalation of hydrogen gas reduces liver injury during major hepatotectomy in swine. World J Gastroenterol 18: 5197-5204. 42. Kajiya M, Sato K, Silva MJ, Ouhara K, Do PM, et al. (2009) Hydrogen from intestinal bacteria is protective for Concanavalin A-induced hepatitis. Biochem Biophys Res Commun 386: 316-321. 43. Liu Q, Shen WF, Sun HY, Fan DF, Nakao A, et al. (2010) Hydrogen-rich saline protects against liver injury in rats with obstructive jaundice. Liver Int 30: 958-968. 44. Sun H, Chen L, Zhou W, Hu L, Li L, et al. (2011) The protective role of hydrogen-rich saline in experimental liver injury in mice. J Hepatol 54: 471-480. 45. Kawai D, Takaki A, Nakatsuka A, Wada J, Tamaki N, et al. (2012) Hydrogen-rich water prevents progression of nonalcoholic steatohepatitis and accompanying hepatocarcinogenesis in mice. Hepatology 56: 912-921. 46. Koyama Y, Taura K, Hatano E, Tanabe K, Yamamoto G, et al. (2013) Effects of oral intake of hydrogen water on liver fibrogenesis in mice. Hepatol Res 44: 663-677. 47. Xia C, Liu W, Zeng D, Zhu L, Sun X, et al. (2013) Effect of hydrogen-rich water on oxidative stress, liver function, and viral load in patients with chronic hepatitis B. Clin Transl Sci 6: 372-375. 48. Lawrence RA, Burk RF (1976) Glutathione peroxidase activity in selenium-deficient rat liver. Biochem Biophys Res Commun 71: 952-958. 49. Bellomo G, Mirabelli F, DiMonte D, Richelmi P, Thor H, et al. (1987) Formation and reduction of glutathione-protein mixed disulfides during oxidative stress. A study with isolated hepatocytes and menadione (2-methyl-1,4-naphthoquinone). Biochem Pharmacol 36: 1313-1320. 50. Aebi H (1984) Catalase in vitro. Methods Enzymol 105: 121-126. 51. Knodell RG, Ishak KG, Black WC, Chen TS, Craig R, et al. (1981) Formulation and application of a numerical scoring system for assessing histological activity in asymptomatic chronic active hepatitis. Hepatology 1: 431-435. 52. Ruwart MJ, Wilkinson KF, Rush BD, Vidmar TJ, Peters KM, et al. (1989) The integrated value of serum procollagen III peptide over time predicts hepatic hydroxyproline content and stainable collagen in a model of dietary cirrhosis in the rat. Hepatology 10: 801-806. 53. Manibusan MK, Odin M, Eastmond DA (2007) Postulated Carbon Tetrachloride Mode of Action: A Review. J Environ Sci Health C Environ Carcinog Ecotoxicol Rev 25: 185-209. 54. Lee HS, Huang GT, Chen CH, Chiou LL, Lee CC, et al. (2001) Less reversal of liver fibrosis after prolonged carbon tetrachloride injection. Hepatogastroenterology 48: 1312-1315. 55. Arthur MJ (1994) Degradation of matrix proteins in liver fibrosis. Pathol Res Pract 190: 825-833. 56. Brenner DA, Waterboer T, Choi SK, Lindquist JN, Stefanovic B, et al. (2000) New aspects of hepatic fibrosis. J Hepatol 32: 32-38. 57. Berry EM, Kohen R (1999) Is the biological antioxidant system integrated and regulated? Med Hypotheses 53: 397-401. 58. Rahman Q, Abidi P, Afaq F, Schiffmann D, Mossman BT, et al. (1999) Glutathione redox system in oxidative lung injury. Crit Rev Toxicol 29: 543-568. 59. Doroshow JH, Locker GY, Myers CE (1980) Enzymatic defenses of the mouse heart against reactive oxygen metabolites: alterations produced by doxorubicin. J Clin Invest 65: 128-135. 60. Senoo H (2004) Structure and function of hepatic stellate cells. Med Electron Microsc 37: 3-15. 61. Stickel F, Brinkhaus B, Krahmer N, Seitz HK, Hahn EG, et al. (2002) Antifibrotic properties of botanicals in chronic liver disease. Hepatogastroenterology 49: 1102-1108. 62. Takahara T, Furui K, Funaki J, Nakayama Y, Itoh H, et al. (1995) Increased expression of matrix metalloproteinase-II in experimental liver fibrosis in rats. Hepatology 21: 787-795. 63. Kumar A, Takada Y, Boriek AM, Aggarwal BB (2004) Nuclear factor-kappaB: its role in health and disease. J Mol Med (Berl) 82: 434-448. 64. Hellerbrand C, Jobin C, Iimuro Y, Licato L, Sartor RB, et al. (1998) Inhibition of NFκB in activated rat hepatic stellate cells by proteasome inhibitors and an IκB super-repressor. Hepatology 27: 1285-1295. 65. Lang A, Schoonhoven R, Tuvia S, Brenner DA, Rippe RA (2000) Nuclear factor kappaB in proliferation, activation, and apoptosis in rat hepatic stellate cells. J Hepatol 33: 49-58. 66. Luedde T, Schwabe RF (2011) NF-κB in the liver—linking injury, fibrosis and hepatocellular carcinoma. Nat Rev Gastroenterol Hepatol 8: 108-118. 67. Marra F, Arrighi MC, Fazi M, Caligiuri A, Pinzani M, et al. (1999) Extracellular signal-regulated kinase activation differentially regulates platelet-derived growth factor's actions in hepatic stellate cells, and is induced by in vivo liver injury in the rat. Hepatology 30: 951-958. 68. Kluwe J, Pradere JP, Gwak GY, Mencin A, De Minicis S, et al. (2010) Modulation of hepatic fibrosis by c-Jun-N-terminal kinase inhibition. Gastroenterology 138: 347-359. 69. Wang Y, Huang Y, Guan F, Xiao Y, Deng J, et al. (2013) Hypoxia-inducible factor-1alpha and MAPK co-regulate activation of hepatic stellate cells upon hypoxia stimulation. PLoS One 8: e74051. 70. Zhao Y, Ma X, Wang J, He X, Hu Y, et al. (2014) Curcumin Protects against CCl4-Induced Liver Fibrosis in Rats by Inhibiting HIF-1α Through an ERK-Dependent Pathway. Molecules 19: 18767. 71. Jazwa A, Cuadrado A (2010) Targeting heme oxygenase-1 for neuroprotection and neuroinflammation in neurodegenerative diseases. Curr Drug Targets 11: 1517-1531. 72. Kawamura T, Wakabayashi N, Shigemura N, Huang C-S, Masutani K, et al. (2013) Hydrogen gas reduces hyperoxic lung injury via the Nrf2 pathway in vivo. Am J Physiol Lung Cell Mol Physiol 304: L646-L656. 73. Trinchet JC, Coste T, Levy VG, Vivet F, Duchatelle V, et al. (1989) [Treatment of alcoholic hepatitis with silymarin. A double-blind comparative study in 116 patients]. Gastroenterol Clin Biol 13: 120-124. 74. Pares A, Planas R, Torres M, Caballeria J, Viver JM, et al. (1998) Effects of silymarin in alcoholic patients with cirrhosis of the liver: results of a controlled, double-blind, randomized and multicenter trial. J Hepatol 28: 615-621. 75. Luper S (1998) A review of plants used in the treatment of liver disease: part 1. Altern Med Rev 3: 410-421. 76. Ono H, Nishijima Y, Adachi N, Tachibana S, Chitoku S, et al. (2011) Improved brain MRI indices in the acute brain stem infarct sites treated with hydroxyl radical scavengers, Edaravone and hydrogen, as compared to Edaravone alone. A non-controlled study. Med Gas Res 1: 12. 77. Nakao A, Kaczorowski DJ, Wang Y, Cardinal JS, Buchholz BM, et al. (2010) Amelioration of rat cardiac cold ischemia/reperfusion injury with inhaled hydrogen or carbon monoxide, or both. J Heart Lung Transplant 29: 544-553. 78. Shinbo T, Kokubo K, Sato Y, Hagiri S, Hataishi R, et al. (2013) Breathing nitric oxide plus hydrogen gas reduces ischemia-reperfusion injury and nitrotyrosine production in murine heart. Am J Physiol Heart Circ Physiol 305: H542-550. 79. Nakashima-Kamimura N, Mori T, Ohsawa I, Asoh S, Ohta S (2009) Molecular hydrogen alleviates nephrotoxicity induced by an anti-cancer drug cisplatin without compromising anti-tumor activity in mice. Cancer Chemother Pharmacol 64: 753-761. 80. Ohta S (2014) Molecular hydrogen as a preventive and therapeutic medical gas: initiation, development and potential of hydrogen medicine. Pharmacol Ther 144: 1-11. 81. Saitoh Y, Harata Y, Mizuhashi F, Nakajima M, Miwa N (2010) Biological safety of neutral-pH hydrogen-enriched electrolyzed water upon mutagenicity, genotoxicity and subchronic oral toxicity. Toxicol Ind Health 26: 203-216. 82. Nakai Y, Sato B, Ushiama S, Okada S, Abe K, et al. (2011) Hepatic oxidoreduction-related genes are upregulated by administration of hydrogen-saturated drinking water. Biosci Biotechnol Biochem 75: 774-776. 83. Kamimura N, Nishimaki K, Ohsawa I, Ohta S (2011) Molecular hydrogen improves obesity and diabetes by inducing hepatic FGF21 and stimulating energy metabolism in db/db mice. Obesity (Silver Spring) 19: 1396-1403. 84. Koyama Y, Taura K, Hatano E, Tanabe K, Yamamoto G, et al. (2014) Effects of oral intake of hydrogen water on liver fibrogenesis in mice. Hepatol Res 44: 663-677. 85. Ostojic SM (2015) Molecular hydrogen: An inert gas turns clinically effective. Ann Med 47: 301-304.
摘要: 
第一部分
杜莎藻在體外試驗中已被證實具有抗氧化能力及誘導癌細胞凋亡,然而,尚無杜莎藻在動物體內的抗血癌活性及免疫活性相關研究。因此,本研究利用Balb/c小鼠尾靜脈注射WEHI-3血癌細胞誘導小鼠血癌動物模式,評估杜莎藻是否具有抗血癌及免疫調節作用。實驗分為7組,分別為正常組、單獨杜莎藻餵食組、血癌誘導組及184.5、369、922.5 mg/kg杜莎藻組,另以主要營養素β-胡蘿蔔素為對照組,杜莎藻以管餵方式每日連續投予二或四週。實驗結果顯示,杜莎藻連續投予血癌小鼠四週期間,可以抑制血癌小鼠肝脾轉移及延長存活率。杜莎藻可以增加血癌小鼠T細胞 (CD3)、B細胞 (CD19) 的比例及降低單核球 (CD11b)、巨噬細胞 (Mac-3) 的比例;脾臟細胞分別以Con A及LPS進行刺激,結果顯示杜莎藻可以促進血癌小鼠T細胞及B細胞增生;另外也發現杜莎藻會增加血癌小鼠自然殺手細胞毒殺能力及巨噬細胞吞噬能力;在免疫調節相關細胞激素分析,杜莎藻會促進血癌小鼠干擾素-γ、介白素 2及減少介白素-4、介白素-10的表現。綜合以上結果可知,杜莎藻可經由免疫調節作用達到抗血癌活性及延長血癌小鼠存活率。

第二部分
氫是一種新穎抗氧化物質,在許多的動物模式結果顯示它可以選擇性的清除強力氧化物及緩解多種氧化壓力相關疾病。本研究進一步以四氯化碳誘導小鼠肝損傷動物模式來評估富氫水的護肝作用,並觀察富氫水是否可以提升水飛薊素的保護效應。ICR雄性小鼠分為6組,每組10隻,分別為正常組、四氯化碳誘導組、水飛薊素治療組、無氫水 (HPW) 組、富氫水 (HRW) 組及合併水飛薊素及富氫水組。四氯化碳誘導八週後會導致肝臟白血球浸潤、肝細胞空泡化、壞死、纖維化及增加血清中ALT、AST活性,並且四氯化碳會顯著性的使肝臟產生脂質過氧化、消耗GSH及降低抗氧化酵素 (SOD、CAT、GSH-Px) 活性,同時也增加發炎反應因子 (TNF-α、IL-6、iNOS、COX-2、HMGB1) 表現,無氫水的給予並沒有產生緩解的效用,而氫水及水飛薊素的給予則可以降低血清中ALT、AST活性和減少肝臟脂質過氧化情形、增加肝臟抗氧化酵素活性、降低TNF-α、IL-6含量及降低iNOS、COX-2、HMGB1表現,特別是合併水飛薊素與氫水處理的組別在各指標上呈現最佳的緩解作用,尤其是在組織病理上能顯著改善肝纖維化情形。進一步在相關信息傳遞路徑上,氫水可以抑制被四氯化碳所活化ERK、AKT、JNK、NF-kB、HIF-1α的蛋白表現,並且降低MMP2、MMP9、HSP70蛋白表現及增加HO-1、γ-GCS蛋白表現。因此,本研究顯示富氫水可以透過清除自由基、提升抗氧化活性及降低發炎反應,並可達到促進水飛薊素對四氯化碳誘導肝發炎及氧化壓力的保護作用。

part 1
Dunaliella salina (D. salina) has been shown to have antioxidant property and induce apoptotic cell death of human cancer cell in vitro. However, there is no information available on anti-leukemia effect or immunomodulatory activity in vivo. In this study, we applied D. salina to syngeneic leukemia-implanted mice (BALB/c and WEHI-3) to investigate its immunological and anti-leukemia properties. Oral administration of D. salina (184.5, 369 and 922.5 mg/kg) for 4 weeks could inhibit tumor metastasis into spleen and liver, and prolonged the survival rate in BALB/c leukemia mice, they were received an intravenous injection of WEHI-3 cells. It had also increased the population and proliferation of T cells (CD3) and B cells (CD19) in splenocytes following Con A/LPS stimulation. Furthermore, D. salina increased the phagocytosis of macrophages and enhanced the cytotoxicity of natural killer cells on flow cytometry assay. Moreover, D. salina enhanced the levels of interferon-γ and interleukin 2 (IL-2) but reduced the levels of IL-4 and IL-10 in leukemia mice. In conclusion, these results demonstrated that the application of D. salina had beneficial effects to WEHI-3 leukemic mice by prolonging survival via modulating the immune responses.

part 2
Hydrogen, a new antioxidant, was reported to selectively reduce the strongest oxidants and alleviates acute oxidative stress in many models. The protective effects of hydrogen-rich water (HRW) against liver damage were evaluated by its attenuation of carbon tetrachloride (CCl4)-induced hepatotoxicity in mice. Male ICR mice were randomly divided into control, CCl4, CCl4 + Silymarin, CCl4 + HPW, CCl4 + HRW, and CCl4 + Silymarin +HRW groups. Treatment with CCl4 for 8 weeks could induce liver lesions included leukocytes infiltration, hepatocyte vacuolization, necrosis, fibrosis and an increase of serum alanine aminotransferase (ALT), and aminotransferase (AST) activity. In addition, CCl4 significantly increased lipid oxidation, decreased the activities of superoxide dismutase (SOD), catalase, glutathione reductase (GSH-Rd) and glutathione peroxidase (GSH-Px), and increased the inflammatory cytokines of TNF-α and IL-6 in the liver. By contrast, Silymarin plus HRW supplement significantly ameliorated the CCl4-induced liver lesions, lowered the serum levels of hepatic enzyme markers (ALT and AST), increased the activities of antioxidant enzyme and reduced the lipid peroxidation and inflammatory markers of TNF-α and IL-6 in liver compared to Silymarin treatment. Consistent with these observations, the proteins expression of iNOS, COX-2, HMGB1, HSP70, HIF-1α inhibited by Silymarin plus HRW treatment. Silymarin plus HRW also caused a significant increase in antioxidant proteins including HO-1, γ-GCS. Moreover, significant decreases of MMP-9, MMP-2, phosphorylated ERK (p-ERK), AKT (p- AKT), JNK (p-JNK) and NF-κB (p-P65) reduced by sylimarin plus HRW treatment as compared to Silymarin treatment. Therefore, the results of this study reveal that HRW can be proposed to enhance the protecting effects by Silymarin on CCl4-induced inflammation and oxidative stress of liver in mice, and the hepatoprotective effect might be correlated with its free radical scavenging effect on enhancing the activity of antioxidant-defense system and inhibiting of inflammatory response.
URI: http://hdl.handle.net/11455/93031
其他識別: U0005-2808201507532300
Rights: 不同意授權瀏覽/列印電子全文服務
Appears in Collections:獸醫學系所

Files in This Item:
File Description SizeFormat Existing users please Login
nchu-104-8098038004-1.pdf12.02 MBAdobe PDFThis file is only available in the university internal network   
Show full item record
 

Google ScholarTM

Check


Items in DSpace are protected by copyright, with all rights reserved, unless otherwise indicated.