Non-parametric methods were applied, as not all parameters were ideally normally distributed. For all statistical

tests, the significance level was set to P < 0.05. Data were analyzed using SPSS for Windows, version 15.0 (SPSS, Inc, Chicago, Ill). Results Performance during the event The main variables controlled during the race are summarized in Table 2. All participants finished the race although two athletes (number 4 and 8 on the Tables 1 to 4) reported gastro-intestinal disturbances during the last hours. All cyclists completed six work efforts, except for two riders who completed seven (subjects number 2 and 5 on the Tables 2 to 5). The mean intensity decreased significantly in riders performing six work efforts MLN2238 (1st work effort: 91 ± 3% of maximum heart rate [HRmax]; 6th work effort: 86 ± 4% of HRmax; P = 0.004) and also those completing seven (1st work effort: 90 ± 5% of HRmax; 7thwork effort: 83 ± 9% of HRmax; P = 0.002) (Figure 1). The mean cumulative climb during the race was 3168 ± 636 m. The cyclists rested between bouts of exercise for 173.2 ± 15.6 min. Table 2 Performance during the event. click here Subjects 1 2 3 4 5 6 7 8 Mean ± SD Racing time (min) 358 406 381 303 495 330 299 318 361 ± 66 Average intensity (% HRmax)

a 88.4 85.3 83.7 90.8 82.4 88.1 87.5 89.8 87.0 ± 2.9 Time spent in zone I (min)b 39 30 63 7 81 56 34 78 49 ± 26 Time spent in zone II (min)b 207 223 225 89 345 111 140 121 183 ± 84 Time spent in zone III (min)b 112 153 93 207 59 163 129 119 129 ± 45 TRIMP 789 935 792 806 948 767 697 677 801 ± 98 Distance (km) 207 223 208 165 282 182 171 163 200 ± 40 Average speed (km/h) 34.7 33.0 32.8 32.7 34.9 33.1 33.9 30.8 33.2 ± 1.3 Recovery time (min) 1082 1034 1059 1137 945 1110 1141 1122 1079 ± 66 a: percentage of maximum heart rate; b: time spent in

each zone of exercise intensity during the racing time (zone I: below to the ventilatory threshold; Thalidomide zone II; between the ventilatory threshold and respiratory compensation point; zone III: above to the respiratory compensation point); TRIMP: training LCZ696 solubility dmso impulse. Figure 1 Evolution of the intensity, expressed as % of maximum heart rate (HR max ), during the event. * Statistical difference (P < 0.05) mean intensity between the first relay compared with the sixth and seventh relay. Macronutrient intake Food and fluids rich in carbohydrates were the main source of energy consumed during the event (Table 3). The athletes consumed 395 ± 193 (5.4 ± 2.6 g/kg; 42 ± 10%, respectively) and 549 ± 141 g of carbohydrates (7.7 ± 2.1 g/kg body mass; 58 ± 10%, respectively) during the first (1900 – 0700 h) and the second (0700 – 1900 h) period, respectively. Carbohydrates reported as fluids and solids were 533 ± 175 g (56.8 ± 10.6%) and 410 ± 174 g (43.2 ± 10.6%), respectively. Protein intake was heterogeneous, while three athletes ingested at rates above 2.5 g/kg body mass; the intake of the remaining subjects were below 2.0 g/kg body mass.