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Glen Davison and Michael Gleeson

The aim of the present study was to investigate the effect of vitamin C with or without carbohydrate consumed acutely in beverages before and during prolonged cycling on immunoendocrine responses. In a single blind, randomized manner six healthy, moderately trained males exercised for 2.5 h at 60% VO2max and consumed either placebo (PLA), carbohydrate (CHO, 6% w/v), vitamin C (VC, 0.15% w/v) or CHO+VC beverages before and during the bouts; trials were separated by 1 wk. CHO and CHO+VC significantly blunted the post-exercise increase in plasma concentrations of cortisol, ACTH, total leukocyte, and neutrophil counts and limited the decrease in plasma glucose concentration and bacteria-stimulated neutrophil degranulation. VC increased plasma antioxidant capacity (PAC) during exercise (P < 0.05) but had no effect on any of the immunoendocrine responses (P > 0.05). CHO+VC increased PAC compared to CHO but had no greater effects, above those observed with CHO alone, on any of the immunoendocrine responses. In conclusion, acute supplementation with a high dose of VC has little or no effect on the hormonal, interleukin-6, or immune response to prolonged exercise and combined ingestion of VC with CHO provides no additional effects compared with CHO alone.

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Tzai-Li Li and Michael Gleeson

This study compared immunoendocrine responses to a single bout of prolonged cycling at different times of day and to a 2nd bout of cycling at the same intensity on the same day. In a counterbalanced design, 8 men participated in 3 experimental trials separated by at least 4 d. In the afternoon exercise-only trial, subjects cycled for 2 h at 60% VO2max starting at 14:00. In the other 2 trials, subjects performed either 2 bouts of cycling at 60% VO2max for 2 h (starting at 09:00 and 14:00) or a separate resting trial. The single bout of prolonged exercise performed in the afternoon induced a larger neutrophilia and monocy-tosis than the identical bout of morning exercise, possibly the result of reduced carbohydrate availability and the circadian rhythm in cortisol levels. The 2nd prolonged exercise bout caused greater immunoendocrine responses but lower plasma glucose levels and neutrophil function compared with the 1st bout.

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John Hough, Caroline Robertson, and Michael Gleeson


This study examined the influence of 10 days of intensified training on salivary cortisol and testosterone responses to 30-min, high-intensity cycling (55/80) in a group of male elite triathletes.


Seven elite male triathletes (age 19 ± 1 y, V̇O2max 67.6 ± 4.5 mL · kg–1 · min–1) completed the study. Swim distances increased by 45%. Running and cycling training hours increased by 25% and 229%, respectively. REST-Q questionnaires assessed mood status before, during, and after the training period. Unstimulated saliva samples were collected before, after, and 30 min after a continuous, high-intensity exercise test. Salivary cortisol and testosterone concentrations were assessed.


Compared with pretraining, blunted exercise-induced salivary testosterone responses to the posttraining 55/80 were found (P = .004). The absolute response of salivary testosterone concentrations to the 55/80 decreased pretraining to posttraining from 114% to 85%. No changes were found in exercise-induced salivary cortisol concentration responses to the 55/80. REST-Q scores indicated no changes in the participants’ psychological stress–recovery levels over the training camp.


The blunted exercise-induced salivary testosterone is likely due to decreased testicular testosterone production and/or secretion, possibly attributable to hypothalamic dysfunction or reduced testicular blood flow. REST-Q scores suggest that the triathletes coped well with training-load elevations, which could account for the finding of no change in the exercise-induced salivary cortisol concentration. Overall, these findings suggest that the 55/80 can detect altered exercise-induced salivary testosterone concentrations in an elite athletic population due to increased training stress. However, this alteration occurs independently of a perceived elevation of training stress.

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Judith Allgrove, Emily Farrell, Michael Gleeson, Gary Williamson, and Karen Cooper

This study investigated the effects of regular consumption of dark chocolate (DC), rich in cocoa polyphenols, on plasma metabolites, hormones, and markers of oxidative stress after prolonged exhaustive exercise. Twenty active men cycled at 60% maximal oxygen uptake (VO2max) for 1.5 hr, with the intensity increased to 90% VO2max for a 30-s period every 10 min, followed by a ride to exhaustion at 90% VO2max. In the 2 wk before exercise participants consumed 40 g of DC or an isocarbohydrate-fat control cocoa liquor–free chocolate (CON) twice daily and once 2 hr before exercise in a randomized, counterbalanced, crossover design. Venous blood samples were taken immediately before exercise, postexercise (fixed duration), postexhaustion, and after 1 hr of recovery. F2-isoprostanes were significantly lower (post hoc tests: p < .001) at exhaustion and after 1 hr of recovery with DC. Oxidized low-density lipoproteins were significantly lower with DC (p < .001) both before and after exercise and at exhaustion. DC was also associated with ~21% greater rises in free fatty acids during exercise (main effect: p < .05). Changes in circulating glucose, insulin, glucagon, cortisol, and interleukin (IL)-6, IL-10, and IL-1ra were unaffected by treatment. Time to exhaustion at 90% VO2max was not significantly different between trials (398 ± 204 and 374 ± 194 s for DC and CON, respectively). These results suggest that regular DC intake is associated with reduced oxidative-stress markers and increased mobilization of free fatty acids after exercise but has no observed effect on exercise performance.

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Michael Gleeson, Nicolette C. Bishop, Marta Oliveira, and Pedro Tauler

The purpose of this study was to examine the effects of a probiotic supplement during 4 mo of winter training in men and women engaged in endurance-based physical activities on incidence of upper respiratory-tract infections (URTIs) and immune markers. Eighty-four highly active individuals were randomized to probiotic (n = 42) or placebo (n = 42) groups and, under double-blind procedures, received probiotic (PRO: Lactobacillus casei Shirota [LcS]) or placebo (PLA) daily for 16 wk. Resting blood and saliva samples were collected at baseline and after 8 and 16 wk. Weekly training and illness logs were kept. Fifty-eight subjects completed the study (n = 32 PRO, n = 26 PLA). The proportion of subjects on PLA who experienced 1 or more weeks with URTI symptoms was 36% higher than those on PRO (PLA 0.90, PRO 0.66; p = .021). The number of URTI episodes was significantly higher (p < .01) in the PLA group (2.1 ± 1.2) than in the PRO group (1.2 ± 1.0). Severity and duration of symptoms were not significantly different between treatments. Saliva IgA concentration was higher on PRO than PLA, significant treatment effect F(1, 54) = 5.1, p = .03; this difference was not evident at baseline but was significant after 8 and 16 wk of supplementation. Regular ingestion of LcS appears to be beneficial in reducing the frequency of URTI in an athletic cohort, which may be related to better maintenance of saliva IgA levels during a winter period of training and competition.

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Nicolette C. Bishop, Michael Gleeson, Ceri W. Nicholas, and Ajmol Ali

Ingesting carbohydrate (CHO) beverages during prolonged, continuous heavy exercise results in smaller changes in the plasma concentrations of several cytokines and attenuates a decline in neutrophil function. In contrast, ingesting CHO during prolonged intermittent exercise appears to have negligible influence on these responses, probably due to the overall moderate intensity of these intermittent exercise protocols. Therefore, we examined the effect of CHO ingestion on plasma interIeukin-6 (IL-6), tumor necrosis factor-α (TNF-α) and lipopolysaccharide (LPS)-stimuIated neutrophil degranulation responses to high-intensity intermittent running. Six trained male soccer players performed 2 exercise trials, 7 days apart, in a randomized, counterbalanced design. On each occasion, they completed six 15-min periods of intermittent running consisting of maximal sprinting interspersed with less intense periods of running and walking. Subjects consumed either CHO or artificially sweetened placebo(PLA) beverages immediately before and at 15-min intervals during the exercise. At 30 min post-exercise, CHO versus PLA was associated with a higher plasma glucose concentration (p< .01), a lower plasma cortisol and IL-6 concentration (p < .02), and fewer numbers of circulating neutrophils (p < .05). Following the exercise, LPS-stimulated elastase release per neutrophil fell 31 % below baseline values on the PLA trial (p = .06) compared with 11% on the CHO trial (p = .30). Plasma TNF-α concentration increased following the exercise (main effect of time, p < .001) but was not affected by CHO. These data indicate that CHO ingestion attenuates changes in plasma IL-6 concentration, neutrophil trafficking, and LPS-stimulated neutrophil degranulation in response to intermittent exercise that involves bouts of very high intensity exercise.

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Alex J. Wadley, Ida S. Svendsen, and Michael Gleeson

Altitude exposure can exaggerate the transient increase in markers of oxidative stress observed following acute exercise. However, these responses have not been monitored in endurance-trained cyclists at altitudes typically experienced while training. Endurance trained males (n = 12; mean (± SD) age: 28 ± 4 years, V̇O2max 63.7 ± 5.3 ml/kg/min) undertook two 75-min exercise trials at 70% relative V̇O2max; once in normoxia and once in hypobaric hypoxia, equivalent to 2000m above sea level (hypoxia). Blood samples were collected before, immediately after and 2 h postexercise to assess plasma parameters of oxidative stress (protein carbonylation (PC), thiobarbituric acid reactive substances (TBARS), total antioxidant capacity (TAC) and catalase activity (CAT)). Participants cycled at 10.5% lower power output in hypoxia vs. normoxia, with no differences in heart rate, blood lactate or rating of perceived exertion observed. PC increased and decreased immediately after exercise in hypoxia and normoxia respectively (nmol/mg/protein: Normoxia—0.3 ± 0.1, Hypoxia + 0.4 ± 0.1; both p < .05). CAT increased immediately postexercise in both trials, with the magnitude of change greater in hypoxia (nmol/min/ml: Normoxia + 12.0 ± 5.0, Hypoxia + 27.7 ± 4.8; both p < .05). CAT was elevated above baseline values at 2 h postexercise in Hypoxia only (Normoxia + 0.2 ± 2.4, Hypoxia + 18.4 ± 5.2; p < .05). No differences were observed in the changes in TBARS and TAC between hypoxia and normoxia. Trained male cyclists demonstrated a differential pattern/ timecourse of changes in markers of oxidative stress following submaximal exercise under hypoxic vs. normoxic conditions.

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Judith E. Allgrove, Louise Geneen, Sarah Latif, and Michael Gleeson

This study investigated the effect of a fed or fasted state on the salivary immunoglobulin A (s-IgA) response to prolonged cycling. Using a randomized, crossover design, 16 active adults (8 men and 8 women) performed 2 hr of cycling on a stationary ergometer at 65% of maximal oxygen uptake on 1 occasion after an overnight fast (FAST) and on another occasion 2 hr after consuming a 2.2-MJ high-carbohydrate meal (FED). Timed, unstimulated whole saliva samples were collected immediately before ingestion of the meal, immediately preexercise, 5 min before cessation of exercise, immediately postexercise, and 1 hr postexercise. The samples were analyzed for s-IgA concentration, osmolality, and cortisol, and saliva flow rates were determined to calculate s-IgA secretion rate. Saliva flow rate decreased by 50% during exercise (p < .05), and s-IgA concentration increased by 42% (p < .05), but s-IgA secretion rate remained unchanged. There was a 37% reduction in s-IgA:osmolality postexercise (p < .05), and salivary cortisol increased by 68% (p < .05). There was no effect of FED vs. FAST on these salivary responses. The s-IgA concentration, secretion rate, and osmolality were found to be significantly lower in women than in men throughout the exercise protocol (p < .05); however, there was no difference between genders in saliva flow rate, s-IgA:osmolality ratio, or cortisol. These data demonstrate that a fed or fasted state 2 hr before exercise does not influence resting s-IgA or the response to prolonged cycling. Furthermore, these results show lower levels of s-IgA and osmolality in women than in men at rest.

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Tzai-Li Li, Ching-Ling Wu, Michael Gleeson, and Clyde Williams

The purpose of this study was to determine the effect of pre-exercise high carbohydrate meals with high glycemic index (HGI) or low glycemic index (LGI) on blood leukocyte redistribution during subsequent endurance exercise. Eight male subjects performed a 90-min run on a treadmill at 70% VO2max 3 h after ingesting an isocaloric HGI or LGI meal with GI values of 77 and 37, respectively. Blood counts of leukocytes, and neutrophils and the neutrophil/lymphocyte ratio were significantly lower in LGI than HGI at 90 min of exercise (P < 0.05). The plasma glucose concentrations were significantly higher in LGI than HGI between 15 min and 45 min of exercise. There were, however, no differences in plasma cortisol, growth hormone, and interleukin-6 concentrations between trials. Thus, the GI of a pre-exercise meal influences leukocyte trafficking and plasma glucose but has limited effects on circulating stress hormone and cytokine responses to exercise.

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Graeme I. Lancaster, Roy L.P.G. Jentjens, Luke Moseley, Asker E. Jeukendrup, and Michael Gleeson

The purpose of the present study was to examine the effect of pre-exercise carbohydrate (CHO) ingestion on circulating leukocyte numbers, plasma interleukin (IL)-6, plasma cortisol, and lipopolysaccharide (LPS)-stimulated neutrophil degranulation responses in moderately trained male cyclists who completed approximately 1-h of high-intensity cycling. The influence of the timing of pre-exercise CHO ingestion was investigated in 8 subjects who consumed 75 g CHO as a glucose solution at either 15 (–15 trial), or 75 (–75 trial) min before the onset of exercise. The influence of the amount of pre-exercise CHO ingestion was investigated in a further 10 subjects who consumed either 25 g or 200 g CHO as a glucose solution or a placebo 45 min before the onset of exercise. At the onset of exercise in the timing experiment, the plasma glucose concentration was significantly (p < .05) lower on the –75 trial compared with pre-drink values, and the plasma cortisol concentration and neutrophil to lymphocyte (N/L) ratio were significantly (p < .05) elevated in the post-exercise period. In the –15 trial, plasma glucose concentration was well maintained, and the plasma cortisol concentration and N/L ratio were not significantly elevated above resting levels. However, LPS-stimulated neutrophil degranulation was similar in the –15 and –75 trials. The amount of CHO ingested had no effect on the magnitude of the rise in the N/L ratio compared with placebo when consumed 45 min pre-exercise. Finally, although an exercise-induced increase in the plasma IL-6 concentration was observed, this effect was independent of pre-exercise CHO ingestion.