The effects of bovine colostrum supplementation on in vivo immunity following prolonged exercise: a randomised controlled trial.

BACKGROUND: Bovine colostrum (COL) has been advocated as a nutritional countermeasure to exercise-induced immune dysfunction, but there is a lack of research with clinically relevant in vivo measures. AIM: To investigate the effects of COL supplementation on in vivo immunity following prolonged exercise using experimental contact hypersensitivity (CHS) with the novel antigen diphenylcyclopropenone (DPCP). METHODS: In a double-blind design, 31 men were randomly assigned to COL (20 g/day) or placebo (PLA) for 58 days. Participants ran for 2 h at 60% maximal aerobic capacity on day 28 and received a primary DPCP exposure (sensitisation) 20 min after. On day 56, participants received a low-dose-series DPCP challenge to elicit recall of in vivo immune-specific memory (quantified by skinfold thickness 24 and 48 h later). Analysis of the dose-response curves allowed determination of the minimum dose required to elicit a positive response (i.e., sensitivity). RESULTS: There was no difference in summed skinfold thickness responses between COL and PLA at 24 h (p = 0.124) and 48 h (p = 0.405). However, sensitivity of in vivo immune responsiveness was greater with COL at 24 h (p < 0.001) and 48 h (p = 0.023) with doses ~ twofold greater required to elicit a positive response in PLA. CONCLUSIONS: COL blunts the prolonged exercise-induced decrease in clinically relevant in vivo immune responsiveness to a novel antigen, which may be a mechanism for reduced illness reports observed in the previous studies. These findings also suggest that CHS sensitivity is highly relevant to host defence.

Post-exercise hot water immersion induces heat acclimation and improves endurance exercise performance in the heat.

We examined whether daily hot water immersion (HWI) after exercise in temperate conditions induces heat acclimation and improves endurance performance in temperate and hot conditions. Seventeen non-heat-acclimatized males performed a 6-day intervention involving a daily treadmill run for 40 min at 65% V̇O2max in temperate conditions (18 °C) followed immediately by either HWI (N = 10; 40 °C) or thermoneutral (CON, N = 7; 34 °C) immersion for 40 min. Before and after the 6-day intervention, participants performed a treadmill run for 40 min at 65% V̇O2max followed by a 5-km treadmill time trial (TT) in temperate (18 °C, 40% humidity) and hot (33 °C, 40% humidity) conditions. HWI induced heat acclimation demonstrated by lower resting rectal temperature (Tre , mean, -0.27 °C, P < 0.01), and final Tre during submaximal exercise in 18 °C (-0.28 °C, P < 0.01) and 33 °C (-0.36 °C, P < 0.01). Skin temperature, Tre at sweating onset and RPE were lower during submaximal exercise in 18 °C and 33 °C after 6 days in HWI (P < 0.05). Physiological strain and thermal sensation were also lower during submaximal exercise in 33 °C after 6 days in HWI (P < 0.05). HWI improved TT performance in 33 °C (4.9%, P < 0.01) but not in 18 °C. Thermoregulatory measures and performance did not change in CON. Hot water immersion after exercise on 6 days presents a simple, practical, and effective heat acclimation strategy to improve endurance performance in the heat.

Repeated muscle damage blunts the increase in heat strain during subsequent exercise heat stress.

PURPOSE: Exercise-induced muscle damage (EIMD) has recently been shown to increase heat strain during exercise heat stress (HS), and represents a risk factor for exertional heat illness (EHI). We hypothesised that a repeated bout of EIMD blunts the increase in rectal temperature (T re) during subsequent endurance exercise in the heat. METHODS: Sixteen non-heat-acclimated males were randomly allocated to EIMD (n = 9) or control (CON, n = 7). EIMD performed a downhill running treatment at -10 % gradient for 60 min at 65 % [Formula: see text]O2max in 20 °C, 40 % RH. CON participants performed the same treatment but at +1 % gradient. Following treatment, participants rested for 30 min, then performed HS (+1 % gradient running for 40 min at 65 % [Formula: see text]O2max in 33 °C, 50 % RH) during which thermoregulatory measures were assessed. Both groups repeated the treatment and subsequent HS 14 days later. Isometric quadriceps strength was assessed at baseline, and 48 h post-treatment. RESULTS: The decrease in leg strength 48 h post-EIMD trial 1 (-7.5 %) was absent 48 h post-EIMD trial 2 (+2.9 %) demonstrating a repeated bout effect. Final T re during HS was lower following EIMD trial 2 (39.25 ± 0.47 °C) compared with EIMD trial 1 (39.59 ± 0.49 °C, P < 0.01), with CON showing no difference. Thermal sensation and the T re threshold for sweating onset were also lower during HS on EIMD trial 2. CONCLUSION: The repeated bout effect blunted the increase in heat strain during HS conducted after EIMD. Incorporating a muscle-damaging bout into training could be a strategy to reduce the risk of EHI and improve endurance performance in individuals undertaking heavy exercise with an eccentric component in the heat.

Evolution of a protein fold in vitro.

A "switch" mutant of the Arc repressor homodimer was constructed by interchanging the sequence positions of a hydrophobic core residue, leucine 12, and an adjacent surface polar residue, asparagine 11, in each strand of an intersubunit beta sheet. The mutant protein adopts a fold in which each beta strand is replaced by a right-handed helix and side chains in this region undergo significant repacking. The observed structural changes allow the protein to maintain solvent exposure of polar side chains and optimal burial of hydrophobic side chains. These results suggest that new protein folds can evolve from existing folds without drastic or large-scale mutagenesis.

Glutamine, exercise and immune function. Links and possible mechanisms.

Glutamine is the most abundant free amino acid in human muscle and plasma and is utilised at high rates by rapidly dividing cells, including leucocytes, to provide energy and optimal conditions for nucleotide biosynthesis. As such, it is considered to be essential for proper immune function. During various catabolic states including surgical trauma, infection, starvation and prolonged exercise, glutamine homeostasis is placed under stress. Falls in the plasma glutamine level (normal range 500 to 750 mumol/L after an overnight fast) have been reported following endurance events and prolonged exercise. These levels remain unchanged or temporarily elevated after short term, high intensity exercise. Plasma glutamine has also been reported to fall in patients with untreated diabetes mellitus, in diet-induced metabolic acidosis and in the recovery period following high intensity intermittent exercise. Common factors among all these stress states are rises in the plasma concentrations of cortisol and glucagon and an increased tissue requirement for glutamine for gluconeogenesis. It is suggested that increased gluconeogenesis and associated increases in hepatic, gut and renal glutamine uptake account for the depletion of plasma glutamine in catabolic stress states, including prolonged exercise. The short term effects of exercise on the plasma glutamine level may be cumulative, since heavy training has been shown to result in low plasma glutamine levels (< 500 mumol/L) requiring long periods of recovery. Furthermore, athletes experiencing discomfort from the overtraining syndrome exhibit lower resting levels of plasma glutamine than active healthy controls. Therefore, physical activity directly affects the availability of glutamine to the leucocytes and thus may influence immune function. The utility of plasma glutamine level as a marker of overtraining has recently been highlighted, but a consensus has not yet been reached concerning the best method of determining the level. Since injury, infection, nutritional status and acute exercise can all influence plasma glutamine level, these factors must be controlled and/or taken into consideration if plasma glutamine is to prove a useful marker of impending overtraining.

Nutritional aspects of immunosuppression in athletes.

The literature suggests that a heavy schedule of training and competition leads to immunosuppression in athletes, placing them at a greater risk of opportunistic infection. There are many factors which influence exercise-induced immunosuppression, and nutrition undoubtedly plays a critical role. Misinterpretation of published data and misleading media reports have lead many athletes to adopt an unbalanced dietary regimen in the belief that it holds the key to improved performance. Some sports have strict weight categories, whilst in others low body fat levels are considered to be necessary for optimal performance or seen as an aesthetic advantage. This leads some athletes to consume a diet extremely low in carbohydrate content which, whilst causing rapid weight loss, may have undesirable results which include placing the athlete at risk from several nutrient deficiencies. Complete avoidance of foods high in animal fat reduces the intake of protein and several fat-soluble vitamins. On the other hand, diets with a very high carbohydrate content are usually achieved at the expense of protein. In addition, anecdotal and media reports have often promoted the supposed performance benefits of certain vitamins and minerals, yet most athletes do not realise that micronutrient supplementation is only beneficial when correcting a deficiency, and to date there is little scientific evidence to substantiate claims that micronutrients act as an ergogenic aid. Moreover, excessive intakes of micronutrients can be toxic. Deficiencies or excesses of various dietary components can have a substantial impact on immune function and may further exacerbate the immunosuppression associated with heavy training loads. This review examines the role of nutrition in exercise-induced immunosuppression and the effect of both excessive and insufficient nutrient intake on immunocompetence. As much of the present literature concerning nutrition and immune function is based on studies with sedentary participants, the need for future research which directly investigates the relationship between exercise, training, immunity and nutrition is highlighted.

The validity of wireless iButtons and thermistors for human skin temperature measurement.

Skin temperature is a fundamental variable in human thermo-physiology, and yet skin temperature measurement remains impractical in most free-living, exercise and clinical settings, using currently available hard-wired methods. The purpose of this study was to compare wireless iButtons and hard-wired thermistors for human skin temperature measurement. In the first of two investigations, iButtons and thermistors monitored temperature in a controlled water bath (range: 10-40 degrees C) and were referenced against a certified, mercury thermometer. In the second investigation, eight healthy males completed three randomized trials (ambient temperature = 10 degrees C, 20 degrees C and 30 degrees C) while both devices recorded skin temperature at rest (in low and high wind velocities) and during cycle-ergometry exercise. The results are as follows. Investigation 1: both devices displayed very high validity correlation with the reference thermometer (r > 0.999). Prior to correction, the mean bias was +0.121 degrees C for iButtons and +0.045 degrees C for thermistors. Upon calibration correction the mean bias for iButtons and thermistors was not significantly different from zero bias. Interestingly, a typical error of the estimate of iButtons (0.043 degrees C) was 1.5 times less than that of thermistors (0.062 degrees C), demonstrating iButtons' lower random error. Investigation 2: the offset between iButton and thermistor readings was generally consistent across conditions; however, thermistor responses gave readings that were always closer to ambient temperature than those given by iButtons, suggesting potential thermistor drift towards environmental conditions. Mean temperature differences between iButtons and thermistors during resting trials ranged from 0.261 degrees C to 1.356 degrees C. Mean temperature differences between iButtons and thermistors during exercise were 0.989 degrees C (ambient temperature = 10 degrees C), 0.415 degrees C (ambient temperature = 20 degrees C) and 0.318 degrees C (ambient temperature = 30 degrees C). Observed error estimates were within the acceptable limits for the skin temperature method comparison, with typical errors <0.3 degrees C, correlation coefficients >0.9 and CV <1% under all conditions. These findings indicate that wireless iButtons provide a valid alternative for human skin temperature measurement during laboratory and field investigations particularly when skin temperature measurement using other currently available methods may prove problematic.

Salivary IgA response to prolonged exercise in a hot environment in trained cyclists.

UNLABELLED: The aim of this study was to determine the effects of prolonged exercise in hot conditions on saliva IgA (s-IgA) responses in trained cyclists. On two occasions, in random order and separated by 1 week, 12 male cyclists cycled for 2 h on a stationary ergometer at 62 (3)% V(.)O(2 max) [194 (4) W; mean (SEM)], on one occasion (HOT: 30.3 degrees C, 76% RH) and on another occasion ( CONTROL: 20.4 degrees C, 60% RH). Water was available ad-libitum. Venous blood samples and 2-min whole unstimulated saliva samples were collected at pre, post and 2 h post-exercise. The s-IgA concentration was determined using a sandwich-type ELISA. Exercising heart rate, rating of perceived exertion, rectal temperature, corrected body mass loss (P<0.01) and plasma cortisol (P<0.05) were greater during HOT. The decrease in plasma volume post-exercise was similar on both trials [HOT: -6.7 (1.1) and CONTROL: -6.6 (1.3)%; P<0.01]. Saliva flow rate decreased post-exercise by 43% returning to pre-exercise levels by 2 h post-exercise (P<0.05) with no difference between trials. Saliva IgA concentration increased post-exercise (P<0.05) with no difference between trials. Saliva IgA secretion rate decreased post-exercise by 34% returning to pre-exercise levels by 2 h post-exercise (P<0.05) with no difference between trials. These data show that a prolonged bout of exercise results in a reduction in s-IgA secretion rate. Additionally, these data demonstrate that performing prolonged exercise in the heat, with ad libitum water intake, does not influence s-IgA responses to prolonged exercise.

Effect of oral glutamine supplementation on human neutrophil lipopolysaccharide-stimulated degranulation following prolonged exercise.

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.

Effects of exercise intensity, duration and recovery on in vitro neutrophil function in male athletes.

The aim of the present study was to compare the effects of exercise at 80% VO2max (resulting in fatigue within 1 h) with more prolonged exercise at a lower work rate (55% VO2max for up to 3 h) on blood neutrophil function and plasma concentrations of cortisol, glutamine and glucose. Eighteen healthy male subjects (mean+/-SD age 22.5+/-3.7 yrs, VO2max 60.1+/-6.6 ml x kg(-1) x min(-1)) cycled on an electrically braked ergometer at 80% VO2max to fatigue (37+/-19 min). On another occasion, separated by at least one week, subjects performed exercise on the same ergometer at 55% VO2max for 3 h or to fatigue, whichever was the sooner. Mean exercise time was 164+/-23 min. The order of the trials was randomised. Both exercise bouts caused significant (p<0.05) elevations of the blood leucocyte count and plasma cortisol concentration and reductions in the in vitro neutrophil degranulation response to bacterial lipopolysaccharide and oxidative burst activity. After exercise at the lower work rate for a longer duration, plasma cortisol concentration was higher, blood leucocyte and neutrophil counts were higher, blood lymphocytes, plasma glucose and indices of neutrophil function were lower than those observed at 80% VO2max. Plasma glutamine only fell significantly during recovery after the more prolonged exercise. We conclude that when exercise is very prolonged, the diminution of innate immune function is greater, or at least as great as that observed after fatiguing exercise at higher work rates. Furthermore, reductions in neutrophil function after exercise at 80% VO2max were not related to changes in the plasma glutamine concentration, although both plasma glutamine and neutrophil function were decreased at 1 and 2.5 h post-exercise in the long duration exercise trial.

Effect of low- and high-carbohydrate diets on the plasma glutamine and circulating leukocyte responses to exercise.

We examined the effects of a low-carbohydrate (CHO) diet on the plasma glutamine and circulating leukocyte responses to prolonged strenuous exercise. Twelve untrained male subjects cycled for 60 min at 70% of maximal oxygen uptake on two separate occasions, 3 days apart. All subjects performed the first exercise task after a normal diet; they completed the second exercise task after 3 days on either a high-CHO diet (75 +/- 8% CHO, n = 6) or a low-CHO diet (7 +/- 4% CHO, n = 6). The low-CHO diet was associated with a larger rise in plasma cortisol during exercise, a greater fall in the plasma glutamine concentration during recovery, and a larger neutrophilia during the postexercise period. Exercise on the high-CHO diet did not affect levels of plasma glutamine and circulating leukocytes. We conclude that CHO availability can influence the plasma glutamine and circulating leukocyte responses during recovery from intense prolonged exercise.

Effect of exercise-induced muscle damage on the blood lactate response to incremental exercise in humans.

Eccentric muscle actions are known to induce temporary muscle damage, delayed onset muscle soreness (DOMS) and muscle weakness that may persist for several days. The purpose of the present study was to determine whether DOMS-inducing exercise affects blood lactate responses to subsequent incremental dynamic exercise. Physiological and metabolic responses to a standardised incremental exercise task were measured two days after the performance of an eccentric exercise bout or in a control (no prior exercise) condition. Ten healthy recreationally active subjects (9 male, 1 female), aged 20 (SD 1) years performed repeated eccentric muscle actions during 40 min of bench stepping (knee high step; 15 steps x min[-1]). Two days after the eccentric exercise, while the subjects experienced DOMS, they cycled on a basket loaded cycle ergometer at a starting work rate of 150 W, with increments of 50 W every 2 min until fatigue. The order of the preceding treatments (eccentric exercise or control) was randomised and the treatments were carried out 2 weeks apart. Two days after the eccentric exercise, all subjects reported leg muscle soreness and exhibited elevated levels of plasma creatine kinase activity (P < 0.05). Endurance time and peak VO2 during cycling were unaffected by the prior eccentric exercise. Minute volume, respiratory exchange ratio and heart rate responses were similar but venous blood lactate concentration was higher (P < 0.05) during cycling after eccentric exercise compared with the control condition. Peak blood lactate concentration, observed at 2 min post-exercise was also higher [12.6 (SD 1.4) vs 10.9 SD (1.3) mM; P < 0.01]. The higher blood lactate concentration during cycling exercise after prior eccentric exercise may be attributable to an increased rate of glycogenolysis possibly arising from an increased recruitment of Type II muscle fibres. It follows that determination of lactate thresholds for the purpose of fitness assessment in subjects experiencing DOMS is not appropriate.

The effects of carbohydrate supplementation on immune responses to a soccer-specific exercise protocol.

The aim of this study was to determine the effect of carbohydrate (CHO) versus placebo (PLA) beverage consumption on the immune and plasma cortisol responses to a soccer-specific exercise protocol in 8 university team soccer players. In a randomized, counterbalanced design, the players received carbohydrate or placebo beverages before, during and after two 90 min soccer-specific exercise bouts (3 days apart) designed to mimic the activities performed and the distance covered in a typical soccer match. Blood and saliva samples were collected before, during and after the exercise protocol. Plasma lactate concentration increased to approximately 4 mmol x l(-1) at 45 and 90 min of exercise in both treatments (P<0.01). Plasma glucose concentration was significantly lower after 90 min of exercise with ingestion of the placebo than the carbohydrate (PLA: 4.57+/-0.12 mmol x l(-1); CHO: 5.49+/-0.11 mmol x l(-1); P<0.01). The pattern of change in plasma cortisol, circulating lymphocyte count and saliva immunoglobulin A secretion did not differ between the carbohydrate and placebo trials. Blood neutrophil counts were 14% higher 1 h after the placebo trial than the carbohydrate trial (PLA: 4.8+/-0.5x10(9) cells x l(-1); CHO: 4.2+/-0.5x10(9) cells x l(-1); P = 0.06), but the treatment had no effect on the degranulation response of blood neutrophils stimulated by bacterial lipopolysaccharide. We conclude that, although previous studies have shown that carbohydrate feeding is effective in attenuating immune responses to prolonged continuous strenuous exercise, the same cannot be said for a soccer-specific intermittent exercise protocol. When overall exercise intensity is moderate, and changes in plasma glucose, cortisol and immune variables are relatively small, it would appear that carbohydrate ingestion has only a minimal influence on the immune response to exercise.

Carbohydrate beverage ingestion and neutrophil degranulation responses following cycling to fatigue at 75% VO2 max.

Carbohydrate (CHO) beverage ingestion appears to influence neutrophil functional responses to prolonged exercise of a fixed duration. The aim of this randomised study was to examine the effect of CHO (5% w/v) beverage ingestion on lipopolysaccharide (LPS)-stimulated neutrophil degranulation responses in nine recreationally active males who cycled at 75% VO2 max until fatigue. On two separate occasions, subjects ingested either placebo (PLA) or CHO beverages before and at 15 min intervals during the exercise. Subjects exercised for 31% longer on the CHO trial compared with the PLA trial (P < 0.05). At fatigue plasma glucose concentration was significantly lower on the PLA trial compared with the CHO trial (P < 0.05). Plasma cortisol concentrations had increased similarly on both trials at this time. A marked neutrophilia was evident at fatigue and throughout the 4 h recovery period, the magnitude of which was similar on both trials. At fatigue LPS-stimulated elastase release per neutrophil had fallen similarly on both trials compared with pre-exercise values (47% and 50% on the PLA and CHO trials, respectively). In conclusion, our results suggest that CHO beverage ingestion has negligible influence on the hormonal, circulating neutrophil and LPS-stimulated neutrophil degranulation responses when exercise is performed to fatigue.

Pre-exercise carbohydrate status and immune responses to prolonged cycling: I. Effect on neutrophil degranulation.

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.

Pre-exercise carbohydrate status and immune responses to prolonged cycling: II. Effect on plasma cytokine concentration.

Ingesting carbohydrate (CHO) beverages during heavy exercise is associated with smaller changes in the plasma concentrations of several cytokines. 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 plasma interleukin (IL)-6, IL-10, and IL-1 receptor antagonist (IL-1ra) responses to prolonged cycling. Seven 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 1 and 2 hours post-TT, plasma concentrations of IL-6 and IL-10 were 2-fold greater on the LOW trial than on the HIGH trial, and peak plasma concentrations of IL-1ra were 9-fold greater on the LOW trial than on the HIGH trial. These findings suggest that pre-exercise CHO status can influence the plasma cytokine response to prolonged cycling.

PDZ-like domains mediate binding specificity in the Clp/Hsp100 family of chaperones and protease regulatory subunits.

ClpX, a molecular chaperone and the regulatory subunit of the ClpXP protease, is shown to contain tandem modular domains that bind to the C-terminal sequences of target proteins in a manner that parallels functional specificity. Nuclear magnetic resonance studies show that these C-terminal sequences are displayed as disordered peptides on the surface of otherwise folded proteins. The ClpX substrate-binding domains are homologous to sequences in other Clp/Hsp100 proteins and are related more distantly to PDZ domains, which also mediate C-terminal specific protein-protein interactions. Conservation of these binding domains indicates that the mode of substrate recognition characterized here for ClpX will be a conserved feature among Clp/Hsp100 family members and a distinguishing characteristic between this chaperone family and the Hsp70 chaperones.

An evolutionary bridge to a new protein fold.

Arc repressor bearing the N11L substitution (Arc-N11L) is an evolutionary intermediate between the wild type protein, in which the region surrounding position 11 forms a beta-sheet, and a double mutant 'switch Arc', in which this region is helical. Here, Arc-N11L is shown to be able to adopt either the wild type or mutant conformations. Exchange between these structures occurs on the millisecond time scale in a dynamic equilibrium in which the relative populations of each fold depend on temperature, solvent conditions and ligand binding. The N11L mutation serves as an evolutionary bridge from the beta-sheet to the helical fold because in the mutant, Leu is an integral part of the hydrophobic core of the new structure but can also occupy a surface position in the wild type structure. Conversely, the polar Asn 11 side chain serves as a negative design element in wild type Arc because it cannot be incorporated into the core of the mutant fold.

No effect of fluid intake on neutrophil responses to prolonged cycling.

Ingesting carbohydrate beverages during prolonged exercise is associated with fewer numbers of circulating neutrophils and attenuated neutrophil functional responses, yet there is little information about the effect of fluid intake alone on immune responses to prolonged exercise. The aim of this study was to examine the influence of regular fluid ingestion compared with no fluid ingestion on plasma cortisol, circulating neutrophil and lipopolysaccharide (LPS)-stimulated neutrophil degranulation responses to prolonged cycling. In a randomized design, nine recreationally active males cycled for 2 h at 65% VO2max on two occasions with either fluid ingestion (lemon-flavoured water, fluid trial) before and during the exercise, or with no fluid intake at all (no fluid trial). Venous blood samples were obtained at rest, immediately after exercise and 1 h after exercise. Immediately after exercise, the plasma cortisol concentration was significantly higher in the no fluid trial than in the fluid trial (592 +/- 62 vs 670 +/- 63 nmol x l(-1), P < 0.05). Circulating numbers of neutrophils increased 4.5-fold (P < 0.01) and LPS-stimulated elastase release per neutrophil decreased 34 +/- 7% (P < 0.01) immediately after exercise; there were no differences between trials. These results suggest that in ambient environmental conditions, fluid ingestion alone has a negligible effect on circulating neutrophil and LPS-stimulated neutrophil degranulation responses to prolonged exercise.