Introduction: Diet and exercise constitute two of the main pillars in preventing and treating cardiovascular diseases. Several studies have focused in analysing the effect of diet and exercise in specific factors, such us hormones, lipids or sugars, but nothing has been found in literature, to our knowledge, analyzing the global effect at a protein level in humans. The present approach has focused on the characterization of the protein profiles detected after a 6 month combination of diet and exercise in healthy individuals, as well as on the identification of the protein changes related. Objectives: 1.Evaluate the combined effect of diet and exercise on the protein profile of healthy individuals. 2.Determine protein profiles related to diet and exercise as potential signatures of physical wellness. 3.Identify protein variations that promote the beneficial effects of diet and exercise in order to better characterize the molecular mechanisms involved. 4.Compare, in the future, the protein profiles obtained here with those related to cardiovascular Material and methods: In total, 53 rugby players were enrolled in this study and included in three groups, one following a hyperproteic diet with low glucemic index, another following a standard Mediterranean diet, and a third group of players without diet. Blood and urine samples were collected on day 0 and 6 months later, after following diet and regular training. Several anthropometric data were also taken, including body weight, mass and length of body segments. Plasma was obtained by centrifugation and both, plasma and urine were aliquoted and stored at −80° C. Protein profiles in both fluids were analysed using a Surface-Enhanced Laser Desorption and Ionization Time of Flight (SELDI-TOF) mass spectrometer, employing a strong anion exchange (Q10) affinity array. For protein identification, a chromatographic approach using a strong anion exchange column (Uno Q) with the same binding conditions employed in SELDI was done, and protein fractions were further separated by SDS-PAGE. Results: SELDI-TOF analysis allowed the detection of protein profiles at time 0 and six months after diet and exercise, reflecting the effect of these parameters in the protein content of plasma and urine. The comparison of these profiles reported significant differences in protein expression (p-values < 0.05) that allowed us to distribute the rugby players in two well defined groups, depending on the weight changes, with clear expression protein patterns in individuals who lost or gained weight. Similar results were seen when comparing muscular changes but not when considering exercise (not shown). Some of the protein peaks with significant differential expression are actually under further analysis in order to be identified, employing a combination of liquid chromatography (anion exchange) and mass spectrometry (MALDI-TOF). Conclusions: SELDI-TOF analysis allowed us to obtain representative protein profiles of the effect of diet, exercise, body weight, muscle and fat variations. The protein peaks with significant expression variations due to the effect of diet and exercise (p-value < 0.05) could be taken as potential markers of vascular wellness, susceptible to be modified with optimal diet and exercise guidelines.