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Neil P. Walsh, Andrew K. Blannin, Nicolette C. Bishop, Paula J. Robson, and Michael Gleeson

Recent studies have shown that neutrophils can utilize glutamine and that glutamine supplementation can improve neutrophil function in postoperative and burn patients. The present study investigated the influence of oral glutamine supplementation on stimulated neutrophil degranulation and oxidative burst activity following prolonged exercise. Subjects, 7 well-trained men, reported to the laboratory following an overnight fast and cycled for 2 hrs at 60% VO2max on two occasions a week apart. They were randomly assigned to either a glutamine or placebo treatment. For both trials, subjects consumed a sugar-free lemon drink at 15-min intervals until 90 minutes, then a lemon flavored glutamine drink (GLN) or sugar-free lemon drink (PLA) was consumed at 15-min intervals for the remaining exercise and the 2-hr recovery period. Venous blood samples were taken pre-, during, and postexercise. Glutamine supplementation had no effect on the magnitude of postexercise leukocytosis, the plasma elastase concentration following exercise (which increased in both trials), or the plasma elastase release in response to bacterial stimulation (which fell in both trials). Neutrophil function assessed by oxidative burst activity of isolated cells did not change following exercise in either trial. These findings therefore suggest that the fall in plasma glutamine concentration does not account for the decrease in neutrophil function (degranulation response) following prolonged exercise.

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Ricardo J.S. Costa, Robert Walters, James L.J. Bilzon, and Neil P. Walsh

The purpose of the study was to determine the effects of carbohydrate (CHO) intake, with and without protein (PRO), immediately after prolonged strenuous exercise on circulating bacterially stimulated neutrophil degranulation. Twelve male runners completed 3 feeding interventions, 1 week apart, in randomized order after 2 hr of running at 75% VO2max. The feeding interventions included a placebo solution, a CHO solution equal to 1.2 g CHO~/kg body mass (BM), and a CHO-PRO solution equal to 1.2 g CHO/kg BM and 0.4 g PRO/kg BM (CHO+PRO) immediately postexercise. All solutions were flavor and water-volume equivalent (12 ml/kg BM). Circulating leukocyte counts, bacterially stimulated neutrophil degranulation, plasma insulin, and cortisol were determined from blood samples collected preexercise, immediately postexercise, and every 30 min until 180 min postexercise. The immediate postexercise circulating leukocytosis, neutrophilia, and lymphocytosis (p < .01 vs. preexercise) and the delayed lymphopenia (90 min postexercise, p < .05 vs. preexercise) were similar on all trials. Bacterially stimulated neutrophil degranulation decreased during recovery in control (23% at 180 min, p < .01 vs. preexercise) but remained above preexercise levels with CHO and CHO+PRO. In conclusion, CHO ingestion, with or without PRO, immediately after prolonged strenuous exercise prevented the decrease in bacterially stimulated neutrophil degranulation during recovery.

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Stewart J. Laing, Samuel J. Oliver, Sally Wilson, Robert Walters, James L.J Bilzon, and Neil P. Walsh

The aim was to investigate the effects of 48 hr of fluid, energy, or combined fluid and energy restriction on circulating leukocyte and lymphocyte subset counts (CD3+, CD4+, and CD8+) and bacterially stimulated neutrophil degranulation at rest and after exercise. Thirteen healthy men (M ± SEM age 21 ± 1 yr) participated in 4 randomized 48-hr trials. During control (CON) participants received their estimated energy (2,903 ± 17 kcal/day) and fluid (3,912 ± 140 ml/day) requirements. During fluid restriction (FR) they received their energy requirements and 193 ± 19 ml/day water to drink. During energy restriction (ER) they received their fluid requirements and 290 ± 6 kcal/day. Fluid and energy restriction (F+ER) was a combination of FR and ER. After 48 hr, participants performed a 30-min treadmill time trial (TT) followed by rehydration (0–2 hr) and refeeding (2–6 hr). Circulating leukocyte and lymphocyte counts remained unchanged for CON and FR. Circulating leukocyte, lymphocyte, CD3+, and CD4+ counts decreased by ~20% in ER and ~30% in F+ER by 48 hr (p < .01), returning to within 0-hr values by 6 hr post-TT. Circulating neutrophil count and degranulation were unaltered by dietary restriction at rest and after TT. In conclusion, a 48-hr period of ER and F+ER, but not FR, decreased circulating leukocyte, lymphocyte, CD3+, and CD4+ counts but not neutrophil count or degranulation. Circulating leukocyte and lymphocyte counts normalized on refeeding. Finally, dietary restriction did not alter circulating leukocyte, lymphocyte, and neutrophil responses to 30 min of maximal exercise.

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Nicolette C. Bishop, Neil P. Walsh, Donna L. Haines, Emily E. Richards, and Michael Gleeson

Ingesting carbohydrate (CHO) beverages during heavy exercise is associated with smaller shifts in numbers of circulating neutrophils and attenuated changes in neutrophil functional responses. The influence of dietary CHO availability on these responses has not been determined. Therefore, the present study investigated the influence of pre-exercise CHO status on circulating neutrophil and lipopolysaccharide (LPS)-stimulated neutrophil degranulation responses to prolonged cycling. Twelve trained male cyclists performed a glycogen-lowering bout of cycling and were randomly assigned to follow a diet ensuring either greater than 70% (HIGH) or less than 10% (LOW) of daily energy intake from CHO for the next 3 days. On day 4, subjects performed an exercise test that comprised cycling for 1 hour at 60% Wmax immediately followed by a time-trial (TT) ensuring an energy expenditure equivalent to cycling for 30 min at 80% Wmax. Subjects repeated the protocol after 7 days, this time following the second diet. The order of the trials was counterbalanced. At TT completion, the HIGH compared with the LOW trial was associated with higher plasma glucose concentration, lower plasma cortisol concentration, and lower circulating neutrophil count. LPS-stimulated neutrophil degranulation per cell fell similarly on both trials. These findings suggest that pre-exercise CHO status influences neutrophil trafficking but not function in response to prolonged cycling.

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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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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.

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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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Ricardo J.S Costa, Samuel J. Oliver, Stewart J. Laing, Robert Walters, James L.J Bilzon, and Neil P. Walsh

The aim of the study was to determine the influence of immediate and 1-hr-delayed carbohydrate (CHO) and protein (PRO) feeding after prolonged exercise on leukocyte trafficking, bacterially stimulated neutrophil degranulation, saliva secretory IgA (S-IgA) responses, and circulating stress hormones. In randomized order, separated by 1 wk, 9 male runners completed 3 feeding interventions after 2 hr of running at 75% VO2max. During control (CON), participants received water (12 ml/kg body mass [BM]) immediately and 1 hr postexercise. During immediate feeding (IF), participants received a CHO-PRO solution equal to 1.2 g CHO/kg BM and 0.4 g PRO/kg BM immediately postexercise and water 1 hr postexercise. During delayed feeding (DF), participants received water immediately postexercise and CHO-PRO solution 1 hr postexercise. Unstimulated saliva and venous blood samples were collected preexercise, immediately postexercise, and every 20 min until 140 min postexercise. No significant interactions were observed for circulating leukocytes and T-lymphocyte subset counts, S-IgA secretion rate, or plasma cortisol, epinephrine, or norepinephrine concentration. Bacterially stimulated neutrophil degranulation decreased during recovery on CON and DF (24% and 31%, respectively, at 140 min; p < .01) but not on IF. Compared with CON, neutrophil degranulation was higher on IF at 100 min postexercise and higher on IF than DF at 80 min and 100 min onward postexercise (p < .05). Ingestion of a CHO-PRO solution immediately after, but not 1 hr after, prolonged strenuous exercise prevented the decrease in neutrophil degranulation but did not alter circulating stress hormone, leukocyte trafficking, or S-IgA responses. Further research should identify the independent effect of different quantities of CHO and PRO ingestion during recovery on neutrophil responses and other aspects of immune function.

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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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Ricardo J.S. Costa, Vera Camões-Costa, Rhiannon M.J. Snipe, David Dixon, Isabella Russo, and Zoya Huschtscha

.g., elastase degranulation and oxidative burst) may play a role in initiating tissue repair ( Peake, 2002 ; Peake et al., 2017 ; Walsh et al., 2011 ), which equate to exercise recovery process similarities. However, certain aspects of neutrophil function are consistently shown to be depressed for several hours after