Antioxidant activity of olive polyphenols in humans: A review

International Journal for Vitamin and Nutrition Research

Volumn 79, Issue 3, 2009, Pages 152-165

  Raederstorff, Daniel ^a  

^a DSM NUTRITIONAL PRODUCTS LTD   (Switzerland)

Author keywords

Antioxidant; Human; Hydroxytyrosol; Olive; Phenolics; Polyphenols

Indexed keywords

ANTIOXIDANT; BIOLOGICAL MARKER; DNA; HYDROXYTYROSOL; ISOPROSTANE; OLIVE OIL; OXIDIZED LOW DENSITY LIPOPROTEIN; PHENOL DERIVATIVE; POLYPHENOL;

ANTIOXIDANT ACTIVITY; BLOOD LEVEL; CARDIOVASCULAR DISEASE; DIETARY INTAKE; DNA DAMAGE; HUMAN; MEDITERRANEAN DIET; OLIVE; OXIDATIVE STRESS; PLASMA; REVIEW; URINE LEVEL;

ANTIOXIDANTS; DIET, MEDITERRANEAN; FEMALE; FLAVONOIDS; HUMANS; LIPOPROTEINS, LDL; MALE; OLEA; OXIDATIVE STRESS; PHENOLS; PLANT OILS;

ANIMALIA; OLEACEAE;

EID: 77949754615     PISSN: 03009831     EISSN: None     Source Type: Journal    
DOI: 10.1024/0300-9831.79.3.152     Document Type: Review

References (86)

1. Trichopoulou, A. and Critselis, E. (2004) Mediterranean diet and longevity. Eur. J. Cancer Prev. 13, 453-456.
2. Knoops, K.T., de Groot, L.C., Kromhout, D., Perrin, A.E., Moreiras-Varela, O., Menotti, A. and van Staveren, W.A. (2004) Mediterranean diet, lifestyle factors, and 10-year mortality in elderly European men and women: the HALE project. JAMA 292, 1433-1439.
3. Mackenbach, J.P. (2007) The Mediterranean diet story illustrates that "why" questions are as important as "how" questions in disease explanation. J. Clin. Epidemiol. 60, 105-109.
4. Estruch, R., Martinez-Gonzalez Miguel, A., Corella, D., Salas-Salvado, J., Ruiz-Gutierrez, V., Covas Maria, I., Fiol, M., Gomez-Gracia, E., Lopez-Sabater Mari, C., Vinyoles, E., Aros, F., Conde, M., Lahoz, C., Lapetra, J., Saez, G. and Ros, E. (2006) Effects of a Mediterranean-style diet on cardiovascular risk factors: a randomized trial. Ann. Intern. Med. 145, 1-11.
5. Esposito, K., Marfella, R., Ciotola, M., Di Palo, C., Giugliano, F., Giugliano, G., D'Armiento, M., D'Andrea, F. and Giugliano, D. (2004) Effect of a Mediterranean-style diet on endothelial dysfunction and markers of vascular inflammation in the metabolic syndrome: a randomized trial. JAMA 292, 1440-1446.
6. Schroeder, H. (2007) Protective mechanisms of the Mediterranean diet in obesity and type 2 diabetes. J. Nutr. Biochem. 18, 149-160.
7. Martinez-Gonzalez, M.A., Fernandez-Jarne, E., Serrano-Martinez, M., Marti, A., Martinez, J.A. and Martin-Moreno, J.M. (2002) Mediterranean diet and reduction in the risk of a first acute myocardial infarction: an operational healthy dietary score. Eur. J. Nutr. 41, 153-160.
8. Scarmeas, N., Stern, Y., Mayeux, R. and Luchsinger, J.A. (2006) Mediterranean diet, Alzheimer disease, and vascular mediation. Arch. Neurol. 63, 1709-1717.
9. Covas, M.-I. (2007) Olive oil and the cardiovascular system. Pharmacol. Res. 55, 175-186.
10. Tripoli, E., Giammanco, M., Tabacchi, G., Di Majo, D., Giammanco, S. and La Guardia, M. (2005) The phenolic compounds of olive oil: structure, biological activity and beneficial effects on human health. Nutr. Res. Rev. 18, 98-112.
11. Tuck, K.L. and Hayball, P.J. (2002) Major phenolic compounds in olive oil: metabolism and health effects. J. Nutr. Biochem. 13, 636-644.
12. Visioli, F., Poli, A. and Galli, C. (2002) Antioxidant and other biological activities of phenols from olives and olive oil. Med. Res. Rev. 22, 65-75.
13. Manna, C., DellaRagione, F., Cucciolla, V., Borriello, A., D'Angelo, S., Galletti, P. and Zappia, V. (1999) Biological effects of hydroxytyrosol, a polyphenol from olive oil endowed with antioxidant activity. Adv. Exp. Med. Biol. 472, 115-130.
14. Blekas, G., Vassilakis, C., Harizanis, C., Tsimidou, M. and Boskou Dimitrios, G. (2002) Biophenols in table olives. J. Agric. Food Chem. 50, 3688-3692.
15. Romero, C., Manuel, B., Yousfi, K., Garcia, P., Garcia, A. and Garrido, A. (2004) Effect of cultivar and processing method on the contents of polyphenols in table olives. J. Agric. Food Chem. 52, 479-484.
16. Bonomini, F., Tengattini, S., Fabiano, A., Bianchi, R. and Rezzani, R. (2008) Atherosclerosis and oxidative stress. Histol. Histopathol. 23, 381-390.
17. Kregel, K.C. and Zhang, H.J. (2007) An integrated view of oxidative stress in aging: basic mechanisms, functional effects, and pathological considerations. Am. J. Physiol. Regul. Integr. Comp. Physiol. 292, R18-R36.
18. Weinbrenner,T., Schroder,H., Escurriol,V., Fito,M., Elosua, R., Vila, J., Marrugat, J. and Covas, M.I. (2006) Circulating oxidized LDL is associated with increased waist circumference independent of body mass index in men and women. Am. J. Clin. Nutr. 83, 30-35.
19. Maritim, A.C., Sanders, R.A. andWatkins, J.B. (2003) Diabetes, oxidative stress, and antioxidants: a review. J. Biochem. Mol. Toxicol. 17, 24-38.
20. Wassmann, S.,Wassmann, K. and Nickenig, G. (2004) Modulation of oxidant and antioxidant enzyme expression and function in vascular cells. Hypertension 44, 381-386.
21. Madamanchi, N.R., Vendrov, A. and Runge, M.S. (2005) Oxidative stress and vascular disease. Arterioscler. Thromb. Vasc. Biol. 25, 29-38.
22. Lapointe, A., Couillard, C. and Lemieux, S. (2006) Effects of dietary factors on oxidation of low-density lipoprotein particles. J. Nutr. Biochem. 17, 645-658.
23. Kaliora, A.C., Dedoussis, G.V. and Schmidt, H. (2006) Dietary antioxidants in preventing atherogenesis. Atherosclerosis 187, 1-17.
24. Johansen, J.S., Harris, A.K., Rychly, D.J. and Ergul, A. (2005) Oxidative stress and the use of antioxidants in diabetes: linking basic science to clinical practice. Cardiovasc. Diabetol. 4, 5-15.
25. Fito, M., de la Torre, R. and Covas, M.I. (2007) Olive oil and oxidative stress. Mol. Nutr. Food Res. 51, 1215-1224.
26. Marrugat, J., Covas, M.I., Fito, M., Schroder, H., Miro-Casas, E., Gimeno, E., Lopez-Sabater,M.C., de la Torre, R. and Farre, M. (2004) Effects of differing phenolic content in dietary olive oils on lipids and LDL oxidation.Arandomized controlled trial. Eur. J. Nutr. 43, 140-147.
27. Visioli, F., Galli, C., Bornet, F., Mattei, A., Patelli, R., Galli, G. and Caruso,D. (2000) Olive oil phenolics are dose-dependently absorbed in humans. FEBS Lett. 468, 159-160.
28. Miro-Casas, E., Covas, M.I., Farre, M., Fito, M., Ortuno, J.,Weinbrenner, T., Roset, P. and de la Torre, R. (2003) Hydroxytyrosol disposition in humans. Clin. Chem. 49, 945-952.
29. Miro-Casas, E., Covas, M.I., Fito, M., Farre-Albadalejo, M., Marrugat, J. and de la Torre, R. (2003) Tyrosol and hydroxytyrosol are absorbed from moderate and sustained doses of virgin olive oil in humans. Eur. J. Clin. Nutr. 57, 186-190.
30. Weinbrenner, T., Fito, M., Farre Albaladejo, M., Saez, G.T., Rijken, P., Tormos, C., Coolen, S., De La Torre, R. and Covas, M.I. (2004) Bioavailability of phenolic compounds from olive oil and oxidative/antioxidant status at postprandial state in healthy humans. Drugs Exp. Clin. Res. 30, 207-212.
31. Vissers, M.N., Zock, P.L., Roodenburg, A.J., Leenen, R. and Katan, M.B. (2002) Olive oil phenols are absorbed in humans. J. Nutr. 132, 409-417.
32. Corona, G., Tzounis, X., Assunta Dessi, M., Deiana, M., Debnam, E.S., Visioli, F. and Spencer, J.P. (2006) The fate of olive oil polyphenols in the gastrointestinal tract: implications of gastric and colonic microflora-dependent biotransformation. Free Radic. Res. 40, 647-658.
33. Edgecombe S. C., Stretch, G., L. and Hayball, P. J. (2000) Oleuropein, an antioxidant polyphenol from olive oil, is poorly absorbed from isolated perfused rat intestine. J. Nutr. 130, 2996-3002.
34. Visioli, F. and Galli, C. (2002) Biological properties of olive oil phytochemicals. Crit. Rev. Food Sci. Nutr. 42, 209-221.
35. Obied, H.K., Allen, M.S., Bedgood, D.R., Prenzler, P.D., Robards, K. and Stockmann, R. (2005) Bioactivity and analysis of biophenols recovered from olive mill waste. J. Agric. Food Chem. 53, 823-837.
36. Covas, M.I., Ruiz Gutierrez, V., de la Torre, R., Kafatos, A., Lamuela Raventos, R.M., Osada, J., Owen, R.W. and Visioli, F. (2006) Minor components of olive oil: Evidence to date of health benefits in humans. Nutr. Rev. 64, S20-S30.
37. Visioli, F., Bellomo, G., Montedoro, G. and Galli, C. (1995) Low density lipoprotein oxidation is inhibited in vitro by olive oil constituents. Atherosclerosis 117, 25-32.
38. Fito, M., Covas, M.I., Lamuela-Raventos, R.M., Vila, J., Torrents, L., de la Torre, C. and Marrugat, J. (2000) Protective effect of olive oil and its phenolic compounds against low density lipoprotein oxidation. Lipids 35, 633-638.
39. Briante, R., Febbraio, F. and Nucci, R. (2004) Antioxidant/prooxidant effects of dietary non-flavonoid phenols on the Cu2+-induced oxidation of human low-density lipoprotein (LDL). Chem. Biodivers. 1, 1716-1729.
40. Franconi, F., Coinu, R., Carta, S., Urgeghe, P. P., Ieri, F., Mulinacci, N. and Romani, A. (2006) Antioxidant effect of two virgin olive oils depends on the concentration and composition of minor polar compounds. J. Agric. Food Chem. 54, 3121-3125.
41. Goya, L., Mateos, R. and Bravo, L. (2007) Effect of the olive oil phenol hydroxytyrosol on human hepatoma HepG2 cells - Protection against oxidative stress induced by tert- butylhydroperoxide. Eur. J. Nutr. 46, 70-78.
42. Manna, C., Galletti, P., Cucciolla, V., Montedoro, G. and Zappia, V. (1999) Olive oil hydroxytyrosol protects human erythrocytes against oxidative damages. J. Nutr. Biochem. 10, 159-165.
43. Manna, C., D'Angelo, S., Migliardi, V., Loffredi, E., Mazzoni, O., Morrica, P., Galletti, P. and Zappia, V. (2002) Protective effect of the phenolic fraction from virgin olive oils against oxidative stress in human cells. J. Agric. Food Chem. 50, 6521-6526.
44. Gutierrez, V.R., de la Puerta, R. and Catalá, A. (2001) The effect of tyrosol, hydroxytyrosol and oleuropein on the non-enzymatic lipid peroxidation of rat liver microsomes. Mol. Cell. Biochem. 217, 35-41.
45. Stupans, I., Kirlich, A., Tuck, K.L. and Hayball, P.J. (2002) Comparison of radical scavenging effect, inhibition of microsomal oxygen free radical generation, and serum lipoprotein oxidation of several natural antioxidants. J. Agric. Food Chem. 50, 2464-2469.
46. Visioli, F., Bellomo, G. and Galli, C. (1998) Free radical-scavenging properties of olive oil polyphenols. Biochem. Biophys. Res. Comm. 247, 60-64.
47. O'Dowd, Y., Driss, F., Dang, P.M., Elbim, C., Gougerot-Pocidalo, M.A., Pasquier, C. and El-Benna, J. (2004) Antioxidant effect of hydroxytyrosol, a polyphenol from olive oil: scavenging of hydrogen peroxide but not superoxide anion produced by human neutrophils. Biochem. Pharmacol. 68, 2003-2008.
48. Carrasco-Pancorbo, A., Cerretani, L., Bendini, A., Segura-Carretero, A., Del Carlo, M., Gallina-Toschi, T., Lercker, G., Compagnone, D. and Fernandez-Gutierrez, A. (2005) Evaluation of the antioxidant capacity of individual phenolic compounds in virgin olive oil. J. Agric. Food Chem. 53, 8918-8925.
49. Schaffer, S., Podstawa, M., Visioli, F., Bogani, P., Muller, W.E. and Eckert, G.P. (2007) Hydroxytyrosolrich olive mill wastewater extract protects brain cells in vitro and ex vivo. J. Agric. Food Chem. 55, 5043-5049.
50. Rietjens, S. J., Bast, A., de Vente, J. and Haenen, G.R. (2007) The olive oil antioxidant hydroxytyrosol efficiently protects against the oxidative stress-induced impairment of the NO bullet response of isolated rat aorta. Am. J. Physiol. Heart Circ. Physiol. 292, H1931-H1936.
51. Visioli, F., Galli, C., Plasmati, E., Viappiani, S., Hernandez, A., Colombo, C. and Sala, A. (2000) Olive phenol hydroxytyrosol prevents passive smoking-induced oxidative stress. Circulation 102, 2169-2171.
52. Visioli, F., Caruso, D., Plasmati, E., Patelli, R., Mulinacci, N., Romani, A., Galli, G. and Galli, C. (2001) Hydroxytyrosol, as a component of olive mill waste water, is dose- dependently absorbed and increases the antioxidant capacity of rat plasma. Free Rad. Res. 34, 301-305.
53. Fki, I., Sahnoun, Z. and Sayadi, S. (2007) Hypocholesterolemic effects of phenolic extracts and purified hydroxytyrosol recovered from olive mill wastewater in rats fed a cholesterol-rich diet. J. Agric. Food Chem. 55, 624-631.
54. Deiana, M., Rosa, A., Corona, G., Atzeri, A., Incani, A., Visioli, F., Paola Melis,M. and Assunta Dessi,M. (2007) Protective effect of olive oil minor polar components against oxidative damage in rats treated with ferric-nitrilotriacetate. Food Chem. Toxicol. 45, 2434-2440.
55. Rietjens, S.J., Bast, A. and Haenen, G.R. (2007) New insights into controversies on the antioxidant potential of the olive oil antioxidant hydroxytyrosol. J. Agric. Food Chem. 55, 7609-7614.
56. Gimeno, E., de la Torre-Carbot, K., Lamuela-Raventos, R.M., Castellote, A.I., Fito, M., de la Torre, R., Covas,M.I. and Carmen Lopez-Sabater,M. (2007) Changes in the phenolic content of low density lipoprotein after olive oil consumption in men. A randomized crossover controlled trial. Br. J. Nutr. 98, 1243-1250.
57. Covas, M.I., Nyyssonen, K., Poulsen, H.E., Kaikkonen, J., Zunft, H.J.F., Kiesewetter, H., Gaddi, A., de la Torre, R., Mursu, J., Baumler, H., Nascetti, S., Salonen, J.T., Fito, M., Virtanen, J. and Marrugat, J. (2006) The effect of polyphenols in olive oil on heart disease risk factors -Arandomized trial. Ann. Intern. Med. 145, 333-341.
58. Covas, M.I., de la Torre, K., Farre-Albaladejo, M., Kaikkonen, J., Fito, M., Lopez-Sabater, C., Pujadas-Bastardes, M.A., Joglar, J., Weinbrenner, T., Lamuela-Raventos, R.M. and de la Torre, R. (2006) Postprandial LDL phenolic content and LDL oxidation are modulated by olive oil phenolic compounds in humans. Free Radic. Biol. Med. 40, 608-616.
59. Salvini, S., Sera, F., Caruso, D., Giovannelli, L., Visioli, F., Saieva, C., Masala, G., Ceroti, M., Giovacchini, V., Pitozzi,V., Galli,C., Romani,A., Mulinacci, N., Bortolomeazzi, R., Dolara, P. and Palli, D. (2006) Daily consumption of a high-phenol extra-virgin olive oil reduces oxidative DNA damage in postmenopausal women. Br. J. Nutr. 95, 742-751.
60. Fito, M., Cladellas, M., de la Torre, R., Marti, J., Alcantara, M., Pujadas-Bastardes, M., Marrugat, J., Bruguera, J., Lopez-Sabater, M.C., Vila, J. and Covas, M.I. (2005) Antioxidant effect of virgin olive oil in patients with stable coronary heart disease: a randomized, crossover, controlled, clinical trial. Atherosclerosis 181, 149-158.
61. Ruano, J., Lopez-Miranda, J., Fuentes, F., Moreno, J.A., Bellido, C., Perez-Martinez, P., Lozano, A., Gomez, P., Jimenez, Y. and Perez Jimenez, F. (2005) Phenolic content of virgin olive oil improves ischemic reactive hyperemia in hypercholesterolemic patients. J. Am. Coll. Cardiol. 46, 1864-1868.
62. Leger, C.L., Carbonneau, M.A., Michel, F., Mas, E., Monnier, L., Cristol, J.P. and Descomps, B. (2005) A thromboxane effect of a hydroxytyrosol-rich olive oil wastewater extract in patients with uncomplicated type I diabetes. Eur. J. Clin. Nutr. 59, 727-730.
63. Weinbrenner, T., Fito, M., de la Torre, R., Saez, G.T., Rijken, P., Tormos, C., Coolen, S., Albaladejo, M.F., Abanades, S., Schroder, H., Marrugat, J. and Covas, M.I. (2004) Olive oils high in phenolic compounds modulate oxidative/antioxidative status in men. J. Nutr. 134, 2314-2321.
64. Visioli, F., Caruso, D., Galli, C., Viappiani, S., Galli, G. and Sala, A. (2000) Olive oils rich in natural catecholic phenols decrease isoprostane excretion in humans. Biochem. Biophys. Res. Comm. 278, 797-799.
65. Moschandreas, J., Vissers, M.N., Wiseman, S., van Putte, K.P. and Kafatos, A. (2002) Extra virgin olive oil phenols and markers of oxidation in Greek smokers: a randomized cross-over study. Eur. J. Clin. Nutr. 56, 1024-1029.
66. Vissers, M.N., Zock, P.L., Wiseman, S.A., Meyboom, S. and Katan, M.B. (2001) Effect of phenol-rich extra virgin olive oil on markers of oxidation in healthy volunteers. Eur. J. Clin. Nutr. 55, 334-341.
67. Machowetz, A., Poulsen, H.E., Gruendel, S., Weimann, A., Fito, M., Marrugat, J., de la Torre, R., Salonen, J.T., Nyyssonen, K., Mursu, J., Nascetti, S., Gaddi, A., Kiesewetter, H., Baumler, H., Selmi, H., Kaikkonen, J., Zunft, H.J., Covas, M.I. and Koebnick, C. (2007) Effect of olive oils on biomarkers of oxidativeDNAstress in Northern and Southern Europeans. FASEB J. 21, 45-52.
68. Visioli, F., Caruso, D., Grande, S., Bosisio, R., Villa, M., Galli, G., Sirtori, C. and Galli, C. (2005) Virgin Olive Oil Study (VOLOS): vasoprotective potential of extra virgin olive oil in mildly dyslipidemic patients. Eur. J. Nutr. 44, 121-127.
69. Oubina, P., Sanchez-Muniz, F.J., Rodenas, S. and Cuesta, C. (2001) Eicosanoid production, thrombogenic ratio, and serum and LDL peroxides in normoand hypercholesterolaemic post-menopausal women consuming two oleic acid-rich diets with different content of minor components. Br. J. Nutr. 85, 41-47.
70. Gleissner, C.A., Leitinger, N. and Ley, K. (2007) Effects of native and modified low-density lipoproteins on monocyte recruitment in atherosclerosis. Hypertension 50, 276-283.
71. Fraley, A.E. and Tsimikas, S. (2006) Clinical applications of circulating oxidized low-density lipoprotein biomarkers in cardiovascular disease. Curr. Opin. Lipidol. 17, 502-509.
72. Galle, J., Hansen-Hagge, T.,Wanner, C. and Seibold, S. (2006) Impact of oxidized low density lipoprotein on vascular cells. Atherosclerosis 185, 219-226.
73. Holvoet, P. (2004) Oxidized LDL and coronary heart disease. Acta Cardiol. 59, 479-484.
74. Holvoet, P., Harris, T.B., Tracy, R.P., Verhamme, P., Newman, A.B., Rubin, S.M., Simonsick, E.M., Colbert, L.H. and Kritchevsky, S.B. (2003) Association of high coronary heart disease risk status with circulating oxidized LDL in the well-functioning elderly: findings from the Health, Aging, and Body Composition study. Arterioscler. Thromb. Vasc. Biol. 23, 1444-1448.
75. Shimada, K., Mokuno, H., Matsunaga, E., Miyazaki, T., Sumiyoshi, K., Miyauchi, K. and Daida, H. (2004) Circulating oxidized low-density lipoprotein is an independent predictor for cardiac event in patients with coronary artery disease. Atherosclerosis 174, 343-347.
76. Nishi, K., Itabe, H., Uno, M., Kitazato, K.T., Horiguchi, H., Shinno, K. and Nagahiro, S. (2002) Oxidized LDL in carotid plaques and plasma associates with plaque instability. Arterioscler. Thromb. Vasc Biol. 22, 1649-1654.
77. Liu, M.L., Ylitalo, K., Salonen, R., Salonen, J.T. and Taskinen, M.R. (2004) Circulating oxidized low-density lipoprotein and its association with carotid intima-media thickness in asymptomatic members of familial combined hyperlipidemia families. Arterioscler. Thromb. Vasc. Biol. 24, 1492-1497.
78. Meisinger, C., Baumert, J., Khuseyinova, N., Loewel, H. and Koenig, W. (2005) Plasma oxidized low-density lipoprotein, a strong predictor for acute coronary heart disease events in apparently healthy, middle-aged men from the general population. Circulation 112, 651-657.
79. Holvoet, P., Lee, D.H., Steffes, M., Gross, M. and Jacobs, D.R., Jr. (2008) Association between circulating oxidized low-density lipoprotein and incidence of the metabolic syndrome. JAMA 299, 2287-2293.
80. Tsimikas, S. (2006) Oxidized low-density lipoprotein biomarkers in atherosclerosis. Curr. Atheroscler. Rep. 8, 55-61.
81. Visioli, F. and Galli, C. (2003) Olives and their production waste products as sources of bioactive compounds. Curr. Top. Nutraceutical Res. 1, 85-88.
82. Dalle-Donne, I., Rossi, R., Colombo, R., Giustarini, D. and Milzani, A. (2006) Biomarkers of oxidative damage in human disease. Clin. Chem. 52, 601-623.
83. Collins, A.R. (2005) Assays for oxidative stress and antioxidant status: applications to research into the biological effectiveness of polyphenols. Am. J. Clin. Nutr. 81, 261S-267S.
84. Fukumoto, M., Shoji, T., Emoto, M., Kawagishi, T., Okuno, Y. and Nishizawa, Y. (2000) Antibodies against oxidized LDL and carotid artery intima-media thickness in a healthy population. Arterioscler. Thromb. Vasc. Biol. 20, 703-707.
85. Chen, H.W., Kuo, C.L., Huang, C.S., Kuo, S.J. and Liu, C.S. (2008) Oxidized low-density lipoproteins, auto-antibodies against oxidized low-density lipoproteins and carotid intima media thickness in a clinically healthy population. Cardiology 110, 252-259.
86. Miller, E.R., 3rd, Erlinger, T.P., Sacks, F.M., Svetkey, L.P., Charleston, J., Lin, P.H. and Appel, L.J. (2005) A dietary pattern that lowers oxidative stress increases antibodies to oxidized LDL: results from a randomized controlled feeding study. Atherosclerosis 183, 175-182.