The quality and quantity of saliva may be important in defending against pathogens transmitted via the buccal cavity. The aim of the present study was to examine the effect of cycling to exhaustion at moderate and high intensity on various salivary parameters and the time course of recovery. Eighteen male subjects of mixed physical fitness took part in the study. Subjects performed two bouts of exercise on separate occasions at least one week apart. Following an overnight fast, subjects cycled on an electrically braked cycle ergometer at a work rate equivalent to 80% VO2max until exhaustion. On another occasion they cycled on the same ergometer at 55% VO2max for 3 h or to fatigue (whichever was sooner). The order of the rides was randomised. Timed, unstimulated saliva samples were collected pre-exercise, during exercise, at cessation of exercise and at 1, 2.5, 5 and 24 h post-exercise. Saliva samples were analysed for IgA, total protein and osmolality. Saliva flow rate was significantly reduced by exercise (P < 0.01). Saliva IgA concentration, secretion rate and ratio to osmolality increased during exercise (P < 0.01). IgA to protein ratio did not change significantly during exercise. Since saliva protein secretion rate increased during exercise (P < 0.01) it appears that correcting for loss of saliva water by expressing IgA relative to protein is misleading. IgA secretion rate and IgA to osmolality ratio are more appropriate measures and neither parameter was lowered by exercise. The results of this study indicate that exercise may detrimentally affect the quantity of saliva produced, but not the quality of saliva. Furthermore, when exercise is to exhaustion, the intensity of the bout does not appear to influence the saliva response. Neither exercise protocol had any long term effect on saliva as all variables recovered within 1 h post-exercise.

A pilot study was undertaken to investigate the effects of the intake of capsules containing the plant sterols and sterolins (BSS:BSSG mixture) on selected immune parameters of volunteers participating in an ultra-marathon in Cape Town, South Africa. Those runners having received active capsules (n=9) showed less neutrophilia, lymphopenia and leukocytosis when compared to their counterparts having received placebo capsules (n=8): the placebo treated individuals showed significant increases in their total white blood cell numbers as well as in their neutrophils (p=0.03 and 0.03 respectively). Furthermore, statistically significant increases within lymphocyte subsets were observed in the runners having received the active capsules: CD3+ cells increased (p=0.02) as did CD4+ cells (p=0.03). In parallel, the BSS:BSSG capsules decreased the plasma level of IL6 in the runners using the active capsules (p=0.08) and significantly decreased the cortisol: DHEAs ratio (p=0.03), suggesting that these volunteers had less of an inflammatory response and were less immune suppressed during the post-marathon recovery period. These findings justify further investigations into the use of the phytosterols to prevent the subtle immunosuppression associated with excessive physical stress.

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 andcirculaling leukocyte responses during recovery from intense prolonged exercise.

11-␤-hydroxysteroid dehydrogenase type 1 catalyzes the conversion of cortisone to hormonally active cortisol and has been implicated in the pathogenesis of a number of disorders including insulin resistance and obesity. The enzyme is a glycosylated membrane-bound protein that has proved difficult to purify in an active state. Extracted enzyme typically loses the reductase properties seen in intact cells and shows principally dehydrogenase activity. The C-terminal catalytic domain is known to contain a disulfide bond and is located within the lumen of the endoplasmic reticulum, anchored to the membrane by a single N-terminal transmembrane domain. We report here the functional expression of the catalytic domain of the human enzyme, without the transmembrane domain and the extreme N terminus, in Escherichia coli. Moderate levels of soluble active protein were obtained using an N-terminal fusion with thioredoxin and a 6xHis tag. In contrast, the inclusion of a 6xHis tag at the C terminus adversely affected protein solubility and activity. However, the highest levels of active protein were obtained using a construct expressing the untagged catalytic domain. Nonreducing electrophoresis revealed the presence of both monomeric and dimeric disulfide bonded forms; however, mutation of a nonconserved cysteine residue resulted in a recombinant protein with no intermolecular disulfide bonds but full enzymatic activity. Using the optimal combination of plasmid construct and E. coli host strain, the recombinant protein was purified to apparent homogeneity by single step affinity chromatography. The purified protein possessed both dehydrogenase and reductase activities with a K m of 1.4 M for cortisol and 9.5 M for cortisone. This study indicates that glycosylation, the N-terminal region including the transmembrane helix, and intermolecular disulfide bonds are not essential for enzyme activity and that expression in bacteria can provide active recombinant protein for future structural and functional studies.

Glutamine is an essential substrate for the proper functioning of cells of the immune system. Falls in plasma glutamine concentration after exercise may have deleterious consequences for immune cell function and render the individual more susceptible to infection. The purpose of the present study was to examine changes in plasma glutamine concentration (measured using a validated enzymatic spectrophotometric method) following an acute bout of intermittent high-intensity exercise. Eight well-trained male games players took part in the study. Subjects reported to the laboratory following an overnight fast and performed a 1-h cycle exercise task consisting of 20 1-min periods at 100% maximal O2 consumption (VO2max) each separated by 2 min of recovery at 30% VO2max. Venous blood samples were taken before exercise and at 5 min, 1 h, 2.5 h, 5 h and 24 h post-exercise. Glutamine was measured by enzymatic spectrophotometric determination of the ammonia concentration before and after treatment of the plasma with glutaminase (EC 3.5.1.2). Plasma glutamine concentration did not fall in the immediate post-exercise period [pre-exercise 681 (23) microM compared with 663 (46) microM at 5 min post-exercise, mean (SEM)], but fell to 572 (35) microM at 5 h post-exercise (P < 0.05 compared with pre-exercise). Plasma lactate concentration rose to 8.8 (1.0) mM at the end of exercise and fell to 1.8 (0.4) mM at 1 h post-exercise, but plasma concentrations of free fatty acids and beta-hydroxybutyrate both rose substantially in the post-exercise period (to 240% and 400% of pre-exercise levels, respectively). The circulating leucocyte count increased significantly during exercise (P < 0.01), continued to increase in the hours following exercise and peaked at 2.5 h post-exercise (mainly due to a neutrophilia). The fall in the plasma glutamine concentration at 5 h post-exercise could be due to increased renal uptake of glutamine, which generally occurs in conditions of metabolic acidosis or due to a greater removal of glutamine from the plasma resulting from the elevated circulating leucocyte count.