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Biomedical subjects

D Keast

Publications and source records attributed to D Keast.

At least 19 recordsLinked to original sources

Cell numbers and in vitro responses of leucocytes and lymphocyte subpopulations following maximal exercise and interval training sessions of different intensities.

In vitro lymphocyte function and the mobilisation of peripheral blood leucocytes was examined in eight trained subjects who undertook an incremental exercise test to exhaustion and a series of interval training sessions. Venous blood samples were obtained before the incremental test, immediately after, and 30, 60, and 120 min after the test. Interval training sessions were undertaken on separate days and the exercise intensities for each of the different sessions were 30%, 60%, 90% and 120% of their maximal work capacity respectively, as determined from the incremental exercise test. There were 15 exercise periods of 1-min duration separated by recovery intervals of 2 min in each session. Venous blood samples were obtained immediately after each training session. Significant increases in lymphocyte subpopulations (CD3+, CD4+, CD8+, CD20+, and CD56+) occurred following both maximal and supramaximal exercise. This was accompanied by a significant decrease in the response of cultures of peripheral blood lymphocytes to Concanavalin A (ConA), a T-cell mitogen. The state of lymphocyte activation in vivo as measured by CD25+ surface antigen was not, however, affected by acute exercise. The total number of lymphocytes, distribution of lymphocyte subpopulations and in vitro lymphocyte response to ConA had returned to pre-exercise levels within half an hour of termination of exercise but serum cortisol concentrations had not begun to fall at this time. There was a significant decrease in the CD4+:CD8+ cell ratio following exercise; this was more the result of increases in CD3-CD8+ cells (CD8+ natural killer cells) than to CD3+CD8+ cells (CD8+ T-lymphocytes). Decreased responsiveness of T-cells to T-cell mitogens, postexercise, may have been the result of decreases in the percentage of T-cells in postexercise mixed lymphocyte cultures rather than depressed cell function. The cause of this was an increase in the percentage of natural killer cells which did not respond to the T-cell mitogen. The results indicated that while a substantial immediate in vitro "immunomodulation" occurred with acute exercise, this did not reflect an immunosuppression but was rather the result of changes in the proportions of reactive cells in mononuclear cell cultures. We have also demonstrated that the degree of the change in distribution of lymphocyte subpopulation numbers and responsiveness of peripheral blood mononuclear cells in in vitro mitogen reactions increased with increasing exercise intensity. Plasma volume changes may have contributed to some of the changes seen in leucocyte population and subpopulation numbers during and following exercise.

CD4-CD8 Ratio

Biological responses to overload training in endurance sports.

Five subjects undertook 10 days of twice daily interval training sessions on a treadmill followed by 5 days of active recovery. On days 1, 6, 11, and 16 the subjects were required to undertake a test of submaximal and maximal work capacity on a treadmill combined with a performance test consisting of a run to exhaustion with the treadmill set at 18 km.h-1 and 1% gradient. Also on these days a pre-exercise blood sample was collected and analysed for a range of haematological, biochemical and immunological parameters. The subjects experienced a significant fall in performance on day 11 which had returned to pretraining levels on day 16. Serum ferritin concentrations were depressed significantly from pretraining concentrations at the conclusion of the recovery period while the expression of lymphocyte activation antigens (CD25+ and HLA-DR+) was increased both after the training phase and the recovery phase. The number of CD56+ cells in the peripheral circulation was depressed at the conclusion of the recovery period. Several parameters previously reported to change in association with overload training failing to reflect the decrease in performance experienced by subjects in this study, suggesting that overtraining may best be diagnosed through a multifactorial approach to the recognition of symptoms. The most important factor to consider may be a decrease in the level of performance following a regeneration period. The magnitude of this decreased performance necessary for the diagnosis of overtraining and the nature of an "appropriate" regeneration period are, however, difficult to define and may vary depending upon the training background of the subjects and the nature of the preceding training. It may or may not be associated with biochemical, haematological, physiological and immunological indicators. Individual cases may present a different range of symptoms and diagnosis of overtraining should not be excluded based on the failure of blood parameters to demonstrate variation. However, blood parameters may be useful to identify possible aetiology in each separate case report of over-training. An outstanding factor to emerge from this study was the difficulty associated with an objective diagnosis of overtraining and this is a possible reason why there have been new accounts of overtraining research in the literature.

Adult

Inhibition in vitro of the replication of murine cytomegalovirus or reovirus type 3 by the glutamine analogue acivicin.

The replication of synchronised cultures of Balb/c L-3T3 fibroblasts was shown to require glutamine at a concentration of at least 0.12 mM, but the replication of murine cytomegalovirus (MCMV) or reovirus type 3 in the cells required at least 0.25 mM glutamine. In the presence of 2 mM glutamine, the glutamine analogue acivicin inhibited DNA synthesis at a minimum concentration of 2 microM in uninfected cells but partially inhibited the replication of MCMV at 0.12 microM and reovirus at 0.03 microM, and totally inhibited MCMV production at 1.0 microM and reovirus at 0.5 microM. Although the therapeutic index in this instance is not great, it does offer encouragement to explore other amino acid analogues as possible broad-spectrum antiviral agents.

3T3 Cells

Glutamine and macrophage function.

The effects of glutamine concentration on the phagocytosis of an opsonized antigen, the synthesis of RNA, and the production of interleukin-1 (IL-1) by macrophages were investigated in vitro. A minimum A minimum of 0.125 mmol/L glutamine was required for a significant increase in phagocytosis of opsonized sheep erythrocytes, compared with that recorded for macrophages cultured in the absence of glutamine. The synthesis of 3H-RNA by macrophages also required 0.125 mmol/L glutamine in the culture medium before it was significantly increased above the levels of control cultures. A minimum of 0.03 mmol/L glutamine was required for the induction of significant levels of IL-1 by lipopolysaccharide (LPS)-stimulated macrophages. Therefore, recent findings suggesting that decreases in plasma glutamine resulting from major burn injury, sepsis, trauma, and surgery may be partly responsible for the associated impairment of immune function now have a basis in both phagocytosis and in modulation of the synthesis of IL-1 (the first cytokine of the interleukin cascade that leads to specific immunity) by macrophages, in addition to the previously established dependency of lymphocytes on external sources of glutamine for their replication.

Analysis of Variance

Periodisation of training stress--a review.

Athletic performance improves as the athlete adapts to progressively increasing training loads. Empirical observations and studies investigating fluctuations in performance indicate that this adaptation occurs during periods of reduced training, termed regeneration periods. Thus it is essential that adequate regeneration time be included in training programmes so that adaptation can be achieved. In order to induce adaptation, heavy periods of training are used to provide a stimulus for adaptive processes to become functional. The literature and anecdotal accounts suggest that the cycling of light, medium, and heavy periods of training is an optimal method for combining the heavy periods of training with the periods of light training needed to allow adaptation and supercompensation.

Adaptation, Physiological

Periodisation and the prevention of overtraining.

It may be essential for the athlete to train in cycles in order to induce optimal improvements and prevent overtraining. Without sufficient recovery time, adaptation may not occur and the athlete may develop the symptoms of overtraining due to continuous and/or excessive exposure to training stress. Training in cycles provides guidelines for the times in the training programme when regeneration should be complete, and therefore the times when the athlete can be screened for overtraining without confusing the fatigue of overload training with that of overtraining. A periodised training structure provides guidelines for conducting research into the mechanisms of training adaptation and overtraining.

Adaptation, Physiological

Acute intensive interval training and T-lymphocyte function.

Immune suppression has been suggested to occur as a result of acute exercise although results of previous studies are variable, possibly due to the failure of some researchers to control exercise intensity and duration. Most of the studies so far have investigated immediate effects after bouts of exercise mainly in subjects undertaking lower body exercise (running or cycling), and the time course of recovery has rarely been determined. We chose two groups of athletes for our studies. One group represented subjects of a range of fitness levels from recreational runners to high-performance runners. The second group represented kayakists with a similar range of fitness levels. Following interval training designed to stress either the lower or upper body anaerobically, we have now shown that upper body exercise (kayaking) induces similar in vitro responses to those described for lower body exercise. There were no differences between the responses of low-fitness versus high-fitness subjects. In addition we have studied the in vitro responses of leukocytes following acute anaerobic exercise over a 24-h recovery period. The results showed that the reduced lymphocyte proliferative response, in vitro, to the T-cell mitogen CONA experienced immediately after exercise returned to normal levels within 2 h of recovery time. This suggests that the reduction in lymphocyte proliferative response is a short transient one.(ABSTRACT TRUNCATED AT 250 WORDS)

Adolescent

Monitoring exercise stress by changes in metabolic and hormonal responses over a 24-h period.

Metabolic and endocrine responses of 14 subjects of varying levels of fitness to an intensive anaerobic interval training session were assessed before exercise and at 2 h, 4 h, 8 h and 24 h postexercise. The endocrine response of the same subjects to a control day, where they were required not to exercise, was also assessed and compared with the values obtained on the interval training day. Uric acid, urea, and creatine phosphokinase concentrations still remained elevated above pre-exercise values 24 h postexercise. Lactate, creatinine, testosterone and cortisol concentrations were significantly elevated above pre-exercise values immediately postexercise but these had reversed by 2 h postexercise. Over the remainder of the recovery period testosterone concentrations remained significantly lower than values measured at similar times on the control day. This was shown to be due directly to a change in testosterone as sex hormone binding globulin concentration remained constant throughout the recovery period. The data indicate that when comparisons of data were made to control (rest) days, imbalances in homeostasis, due to intensive training, are not totally reversed within the next 24-h. The data also demonstrate that the parameters measured undergo the same variations in subjects with a wide range of physical fitness, indicating that these parameters could be used to monitor exercise stress and recovery in athletes of a wide range of abilities.(ABSTRACT TRUNCATED AT 250 WORDS)

Adolescent

Maximal activities of glutaminase, citrate synthase, hexokinase, 6-phosphofructokinase and lactate dehydrogenase in skin of immune-competent Balb/c and immune-deficient Balb/c (nu/nu) mice during wound healing.

1. The maximal activities of hexokinase (HK), 6-phosphofructokinase (PFK), lactate dehydrogenase, citrate synthase (CS) and glutaminase (GLU) which provide quantitative and qualitative indices of flux through several important metabolic pathways have been examined in the wounded skin of haired immune competent Balb/c mice and hairless immune deficient Balb/c (nu/nu) mice of various ages during the first ten days of wound healing. 2. The potential for glucose utilization and for aerobic metabolism as suggested by the maximal activities of HK, PFK, CS, were raised in the skin of Balb/c mice of various ages on all post wounding days. Increases in the maximal activity of GLU was observed only in the skin of 6 and 10 weeks old Balb/c mice during wound healing. 3. There was no evidence of a contribution to the maximal activity of GLU by infiltrating cells of the immune system to the wound site in the skin of either haired or hairless mice.

Aging

Effect of B- and T-cell mitogens on the maximum activities of hexokinase, lactate dehydrogenase, citrate synthase and glutaminase in bone marrow cells and thymocytes of the rat during four hours of culture.

1. Cells from the bone marrow and cells from the thymus of the rat were incubated in the presence of glucose and glutamine and phytohaemagglutinin, concanavalin-A or lipopolysaccharide. Cells were harvested at times up to 4 hr, extracted and maximum activities of hexokinase, lactate dehydrogenase, citrate synthase or glutaminase measured. 2. In bone marrow cells, there were little changes in enzyme activities except for an increase in the activity of citrate synthase which was prevented by concanavalin-A. This mitogen also caused a decrease in the activity of hexokinase. 3. In contrast, in thymocytes, the activities of hexokinase and glutaminase were decreased in the control condition but addition of lipopolysaccharide, a B-cell mitogen prevented these decreases in activity and concanavalin-A maintained the activity of glutaminase. Concanavalin-A caused a decrease in hexokinase activity but a marked increase in that of glutaminase. 4. It is suggested that changes in the maximum activities of hexokinase and glutaminase over this 4 hr period may represent the effect of removal of thymus-produced growth factors, whose effects can be replaced, at least in part, by two mitogens.

Animals

The effects of diet on the maximal activities of glutaminase, citrate synthase, hexokinase, 6-phosphofructokinase and lactate dehydrogenase in the skin of haired and hairless mice of various ages.

1. The maximal activities of hexokinase (HK), 6-phosphofructokinase (PFK), lactate dehydrogenase (LDH), citrate synthase (CS) and glutaminase (GLU) which provide a quantitative indices of flux through several important pathways have been measured in the skin of haired Balb/c and hairless Balb/c (nu/nu) mice under normal and dietary stress. 2. The skin of old haired mice exhibited higher PFK and LDH activities with lower HK, CS and GLU activities. All activities of enzymes associated with energy metabolism in the skin of old hairless mice were higher than those in the skin of haired mice. 3. HK, LDH, CS and GLU activities were maintained at normal levels in the skin of haired mice when these mice were fed diets deficient in energy or protein components (HPLE, LPNE). These enzymes however were severely suppressed when mice were fed a diet deficient in both energy and protein components (LPLE). Recovery of activities of these enzymes to the control level was observed when mice were refed with the normal diet for a week.

Aging

The effects of a leukaemia-controlling dose of acivicin on murine splenic lymphocytes in vitro and in vivo.

The development of successful chemotherapy for cancers, including leukaemia, is based on the exploitation of significant toxic differentials of the agents, between the cancer cells and their normal counterparts. A concentration of 0.1 mumol/L of the amino acid analogue acivicin (L-(alpha S,5S)-alpha-amino-3-chloro-4,5-dihydro-5-isoxazoleacetic acid) was sufficient to inhibit [3H]-DNA synthesis in mitogen stimulated murine splenic lymphocytes and WEHI 7.1 murine leukaemic cells in vitro, at glutamine concentrations up to 0.5 mmol/L. However, at concentrations of glutamine of 1.0 mmol/L and above, more acivicin was required to inhibit WEHI 7.1 cells than to inhibit the splenic lymphocytes. At 1.0 mmol/L glutamine, 18 h of exposure to an optimal inhibitory concentration of acivicin (0.5 mumol/L) in vitro was sufficient to inhibit [3H]-DNA synthesis in the mitogen stimulated lymphocytes, whereas 24 h of exposure to an optimal inhibitory acivicin concentration of 2.0 mumol/L was required for inhibition of [3H]-DNA synthesis in the WEHI 7.1 leukaemic cells. When an acivicin inoculation regime that was sufficient to control the growth of intraperitoneally (i.p.) implanted WEHI 7.1 leukaemic cells was administered to Balb/c mice, the primary immune response was significantly inhibited to an antigen given either during or after the acivicin treatment. These results indicate that lymphocytes replicating in vivo in a specific primary immune response could be as sensitive to acivicin as leukaemic cells.

Animals

Overtraining in athletes. An update.

Overtraining appears to be caused by too much high intensity training and/or too little regeneration (recovery) time often combined with other training and nontraining stressors. There are a multitude of symptoms of overtraining, the expression of which vary depending upon the athlete's physical and physiological makeup, type of exercise undertaken and other factors. The aetiology of overtraining may therefore be different in different people suggesting the need to be aware of a wide variety of parameters as markers of overtraining. At present there is no one single diagnostic test that can define overtraining. The recognition of overtraining requires the identification of stress indicators which do not return to baseline following a period of regeneration. Possible indicators include an imbalance of the neuroendocrine system, suppression of the immune system, indicators of muscle damage, depressed muscle glycogen reserves, deteriorating aerobic, ventilatory and cardiac efficiency, a depressed psychological profile, and poor performance in sport specific tests, e.g. time trials. Screening for changes in parameters indicative of overtraining needs to be a routine component of the training programme and must be incorporated into the programme in such a way that the short term fatigue associated with overload training is not confused with the chronic fatigue characteristic of overtraining. An in-depth knowledge of periodisation of training theory may be necessary to promote optimal performance improvements, prevent overtraining, and develop a system for incorporating a screening system into the training programme. Screening for overtraining and performance improvements must occur at the culmination of regeneration periods.

Adaptation, Physiological

Effect of mitogens on the maximum activities of hexokinase, lactate dehydrogenase, citrate synthase and glutaminase in rat mesenteric lymph node lymphocytes and splenocytes during the early period of culture.

1. The activities of hexokinase, lactate dehydrogenase and citrate synthase were maintained in mesenteric lymph node lymphocytes during 4 hr of culture: the activity of glutaminase increased during this period of time. 2. In splenocytes, the activity of hexokinase decreased markedly during the 4 hr period, whereas those of lactate dehydrogenase and glutaminase remained constant, and that of citrate synthase increased dramatically. 3. In both mesenteric lymphocytes and splenocytes, addition of the T-cell mitogens, phytohaemagglutinin or concanavalin-A, to the culture medium caused decreases in the activities of both hexokinase and citrate synthase. 4. In contrast, these mitogens increased the activity of glutaminase in both cell types. 5. Addition of the B-cell mitogen, bacterial lipopolysaccharide, had little effect on hexokinase, lactate dehydrogenase or citrate synthase but increased markedly that of glutaminase in mesenteric lymph node lymphocytes. 6. In splenocytes this mitogen prevented much of the decrease in hexokinase activity, increased the activities of citrate synthase and glutaminase but had little effect on that of lactate dehydrogenase.

Animals

The effect of glutamine on murine splenic leukocyte responses to T and B cell mitogens.

Murine splenic leukocytes cannot be stimulated to synthesize [3H]-DNA by various concentrations of either the T cell mitogens, Concanavalin-A (Con-A) or phytohaemagglutinin (PHA) or the B cell mitogen, lipopolysaccharide (LPS), unless glutamine is present in the culture medium. The optimum concentration of the T cell mitogens (Con-A, PHA) remained constant for all levels of glutamine while that of the B cell mitogen (LPS) increased as the concentration of glutamine in the medium increased.

Animals

Fine body movements and the cardiac cycle in archery.

This study examined the relationships between postural sway, aiming time, the cardiac cycle time and the placement of the first finger movement within the electrocardiac cycle, with the quality of the arrow shot. A small group of elite male and female archers who shot either the recurve or the compound bow, freestyle or bare bow was used in this study. A total of 240 arrows were shot, classified as being of good, average or bad quality and analysed in detail. The analyses were undertaken over two occasions up to 100 days apart. The area of postural sway, as measured by the movement of the centre of pressure coordinates, varied significantly (P less than 0.05-0.001) both within and between trials for all archers and for all quality of arrows shot. There was a tendency for the area of postural sway to increase as the quality of the arrow shot decreased. This increase in postural sway exceeded, for arrows of bad quality, that exhibited as normal postural sway when standing relaxed and addressing the target with the hands at the side. Aiming time was variable within the quality of arrow shot, although it was consistent for each archer. In some cases the aiming time increased as the quality of the arrow decreased, whereas in the case of bare bow archery the aiming time tended to decrease as the quality of the arrow deteriorated. The cardiac cycle time increased significantly (P less than 0.05-0.01) as the archers approached the loose of the good quality arrows but there was no significant increase in cardiac cycle time when arrows of average or bad quality were shot. The most consistent parameter related to the quality of the shot was the placement of the first finger movement within the ST phase or the mid-cycle phase of the electrocardiogram for arrows of good quality (P less than 0.0001).

Adolescent