1. Haemostatic cardiovascular risk factors: complementary effects of red wine and mediterranean diet in young population. Diego Mezzano, M.D. Department of Hematology-Oncology School of Medicine P. Catholic University of Chile Talca, Agosto 22, 2006

2. Objectives 1.To compare the effect of an alcohol-free Mediterranean-type diet and a High-fat diet on haemostatic variables. 2.To test whether red wine supplementation modifies these variables, independently of diet.

3. Composition of Diets MDHFD Total calories (kcal/day) 25652565 Calories in protein (%) 17.617.6 Calories in fat (%) 27.339.9 Saturated fats (g/day) 22.735.8 MUFA (g/day) 37.935.5 PUFA (g/day) 9.632.0  -3 FA (g/day) 1.640.96 Olive oil (mL/day) 320 Fruits, vegetables (g/day) 675246

11. MD MD + RED WINE MD HFD HFD + RED WINE HFD 0 30 60 90 GROUP MD GROUP HFD N = 21 Diet Intervention Design

12. Comparison between MD and HFD groups at T0 Variable:MDHFD p = Age (years) 21.2  1.7 22.6  4.5 0.21 Body mass index (kg/m 2 ) 23.4  2.0 23.7  2.5 0.57 Serum albumin (g/L) 5.1  0.24 5.1  0.17 0.82 Blood hemoglobin (g/L) 151  7.5 152  8.7 0.60 Plasma cholesterol (mmol/L) 4.42  0.72 4.57  0.67 0.44 Triacylglycerides (mmol/L) 1.13  0.57 1.05  0.56 0.59

13. Comparison between MD and HFD groups at T0 Variable:MDHFD p = Plasma  -carotene (  mol/L) 0.26  0.15 0.37  0.25 0.08 Plasma ascorbate (  mol/L) 54.7  15.0 53.5  10.9 0.78 Serum folate (nmol/L) 13.8  3.1 11.7  2.8 0.19 Eicosapentaenoic acid (%) 1.20  0.56 0.93  0.44 0.19

14. Comparison between MD and HFD groups after 30 days Variable:MDHFD p = Plasma  -carotene (  mol/L) 0.49  0.19 0.23  0.14 0.0001 Plasma ascorbate (  mol/L) 55.9  11.2 29.7  8.9 0.0001 Serum folate (nmol/L) 14.2  4.3 10.5  2.6 0.002 Eicosapentaenoic acid (%) 1.50  0.7 0.85  0.30 0.001

16. Differential effects of diet and red wine on haemostatic CVRF Several haemostasis proteins which participate in the generation, inhibition or removal of fibrin, are either established or potential CVRF.Several haemostasis proteins which participate in the generation, inhibition or removal of fibrin, are either established or potential CVRF. Among them, plasma levels of fibrinogen, factors VIIc and VIIIc, tPA and PAI-1.Among them, plasma levels of fibrinogen, factors VIIc and VIIIc, tPA and PAI-1.

17. P=0.03 P=0.001 P=0.034 P=0.05 P=0.006 P=0.01

18. p = 0.013

19. p = 0.01

20. p = 0.0003

21. Differential effects of diet and red wine on primary hemostasis variables Primary hemostasis: processes that mediate interaction of platelets with the vessel wall.Primary hemostasis: processes that mediate interaction of platelets with the vessel wall. Platelet function is an essential phenomenon in the pathogenesis of arterial cardiovascular events, and pharmacological agents that decrease platelet-vessel wall interaction are pivotal in preventing and/or treating these accidents.Platelet function is an essential phenomenon in the pathogenesis of arterial cardiovascular events, and pharmacological agents that decrease platelet-vessel wall interaction are pivotal in preventing and/or treating these accidents. Some important variables of primary hemostasis include bleeding time, plasma vWF levels and platelet aggregation and secretion.Some important variables of primary hemostasis include bleeding time, plasma vWF levels and platelet aggregation and secretion.

22. p = 0.017

24. ADP 4  M p = 0.07

25. ADP 8  M

26. p = 0.003 Collagen, 1  g/mL

27. p = 0.005 p = 0.013 Collagen, 2  g/mL

28. Conclusions. 1. 1.Intake of a MD, compared with a HFD, was associated with an overall improvement in haemostatic CV risk factors: lower plasma fibrinogen and factors VIIc and VIIIc, and higher levels of protein S. Red wine supplementation in both diets resulted in further decrease in plasma fibrinogen and factor VIIc, but in increases in PAI-1 and t-PAAg. 2.The longer BT in individuals on MD, independently of red wine, denotes less interaction of platelets with the vascular wall, which could be beneficial from the point of view of cardiovascular (CV) risk.

29. Conclusions. 2. 3.Moderate consumption of red wine results in a significant increase in ex vivo platelet aggregation and secretion after stimulation with collagen, whereas no significant changes were observed when platelets were challenged with other agonists. 4.Our observations denote that some of the beneficial effects of moderate red wine consumption would be mediated by its favourable influence on hemostatic CV risk factors and not on inhibition of platelet responses.

30. Complementary effects of Mediterranean diet and moderate red wine intake on haemostatic cardiovascular risk factors. D, Leighton F, Martínez C, Marshall G, Cuevas A, Castillo O, Panes O, Muñoz B, Pérez DD, Mizón C, Rozowski J, San Martín A, Pereira J. Eur J Clin Nutr 2001; 55: 444-451. Complementary effects of Mediterranean diet and moderate red wine intake on haemostatic cardiovascular risk factors. Mezzano D, Leighton F, Martínez C, Marshall G, Cuevas A, Castillo O, Panes O, Muñoz B, Pérez DD, Mizón C, Rozowski J, San Martín A, Pereira J. Eur J Clin Nutr 2001; 55: 444-451. Mediterranean diet, but not red wine, is associated with beneficial changes on primary haemostasis. Mezzano D, Leighton F, Strobel P, Martínez C, Marshall G, Cuevas A, Castillo O, Panes O, Muñoz B, Rozowski J, Pereira J. Eur J Clin Nutr 2003; 57: 439-446.

32. p = 0.03 p = 0.001

33. p = 0.034 p = 0.05

34. p = 0.006

35. p = 0.01

37. p = 0.02 Epinephrin 8  M