Lymphocyte function and cytokine production during incremental exercise in active and sedentary males and females.

This study examined the effects of acute continuous incremental exercise on lymphocyte mitogenic function and cytokine production in physically active and sedentary males and females. Physically active (n = 32) and sedentary (N = 32) male and female subjects were randomly assigned to an exercise or control condition. Exercise involved a continuous incremental protocol consisting of cycling for 3 periods of 6 min at workrates corresponding to 55%, 70% and 85% VO2peak. Blood samples were drawn from a venous catheter at baseline, 6 min, 12 min and 18 min, and 2 h following completion of exercise. Relative to baseline and control condition the percentage of T (CD3+) and B cells (CD19+) significantly decreased, and the percentage of NK cells (CD3-CD16+CD56+) increased (p < 0.001) during each stage of the incremental exercise test. The proliferative response to ConA was suppressed, enhanced, or unchanged using 1.25 micrograms/ml, 2.5 micrograms/ml and 5.0 micrograms/ml ConA, respectively. The in-vitro production of IL-1 and IFN-gamma increased during each workload. In contrast IL-4 production did not change during exercise. The resting and exercise induced alterations in lymphocyte function and cytokine production were independent of gender and fitness level, and returned to baseline 2 h into recovery. The in-vitro production of IFN-gamma and IL-4 suggests that physical activity may alter the balance of TH1 and TH2 lymphocytes.

The effects of incremental submaximal exercise on circulating leukocytes in physically active and sedentary males and females.

To study the effects of exercise on circulating leukocytes and leukocyte subsets, physically active (n = 32) and sedentary (n = 32) male and female subjects were randomly assigned to an exercise or control condition. Exercise involved a continuous incremental protocol consisting of cycling for three periods of 6 min at power outputs corresponding to 55%, 70% and 85% maximal oxygen uptake (VO2max). Blood samples were drawn from a venous catheter at baseline, and at 6 min, 12 min, and 18 min after beginning the exercise and 2 h following completion of exercise. Resting- and exercise-induced alterations in total leukocytes were independent of gender and subject fitness level. Relative to baseline, each increment in workload resulted in a rapid increase in the number of circulating leukocytes. Increases in neutrophils, lymphocytes and monocytes accounted for the exercise-induced leukocytosis. With regard to lymphocytes, exercise resulted in a significant increase in the number of T cells (CD3+), T helper cells (CD4+), T suppresser (CD8+) and natural killer (NK) cells (CD3-/CD16+/CD56+). The largest percentage increase occurred in the NK cell population. The CD4+: CD8+ ratio decreased (P < 0.001) throughout exercise due to a larger increase in the number of CD8+ cells relative to CD4+ cells. An exercise-induced neutrophilia, lymphocytopenia, and eosinophelia was observed 2 h into recovery. Exercise resulted in significant increases in plasma epinephrine and norepinephrine levels. There was no indication of a hypothalamic-pituitarty-adrenal response during exercise. The results indicate that the rapid, albeit transient, alteration in the number of circulating leukocytes during and following an acute progressive incremental exercise test are independent of gender and fitness.

Exercise-induced alterations in natural killer cell number and function.

To study the effects of exercise on natural killer (NK) cell number and activity (NKCA) healthy male (n = 32) and female (n = 32) subjects were randomly assigned to an exercise or control condition. Exercise involved a continuous incremental protocol consisting of cycling for three periods of 6 min at work rates corresponding to 55%, 70% and 85% peak oxygen uptake (VO2peak). Blood samples were drawn at baseline, at 6 min, 12 min and 18 min during exercise, and at 2 h following completion of exercise. Relative to both baseline and control conditions, exercise resulted in an increase in the number of circulating lymphocytes. The proportion of T cells (CD3+) and B cells (CD19+) significantly decreased, and NK cells (CD3-CD16+CD56+) increased throughout exercise. NKCA increased (P < 0.001) during the initial 6 min of exercise with no further changes observed, despite increases (P < 0.001) in the number and proportion of circulating NK cells during exercise at 70% and 85% VO2peak. Plasma epinephrine and norepinephrine increased (P < 0.001) above baseline at 12 min and 18 min. The changes in NK cell number and function were independent of gender. The results indicate that short-duration low-intensity exercise can significantly increase NK cell number and activity. However, alterations in NK cell number are not accompanied by changes of a similar magnitude in NKCA.

Synaptosomal membrane-bound form of endopeptidase-24.15 generates Leu-enkephalin from dynorphin1-8, alpha- and beta-neoendorphin, and Met-enkephalin from Met-enkephalin-Arg6-Gly7-Leu8.

Brain contains a membrane-bound form of endopeptidase-24.15, a metalloendopeptidase predominantly associated with the soluble protein fraction of brain homogenates. Subcellular fractionation of the enzyme in rat brain showed that 20-25% of the total activity is associated with membrane fractions including synaptosomes. Solubilization of the enzyme from synaptosomal membranes required the use of detergents or treatment with trypsin. The specific activity of the enzyme in synaptosomal membranes measured with tertiary-butoxycarbonyl-Phe-Ala-Ala-Phe-p-aminobenzoate as substrate was higher than that of endopeptidase-24.11 ("enkephalinase"), a membrane-bound zinc-metalloendopeptidase believed to function in brain neuropeptide metabolism. Purified synaptosomal membranes converted efficiently dynorphin1-8, alpha- and beta-neoendorphin into leucine enkephalin and methionine-enkephalin-Arg6-Gly7-Leu8 into methionine enkephalin in the presence of captopril, bestatin, and N-[1-(R,S)-carboxy-2-phenylethyl]-Phe-p-aminobenzoate, inhibitors of angiotensin converting enzyme (EC 3.4.15.1), aminopeptidase (EC 3.4.11.2), and membrane-bound metalloendopeptidase (EC 3.4.24.11), respectively. The conversion of enkephalin-containing peptides into enkephalins was virtually completely inhibited by N-[1-(R,S)-carboxy-2-phenylethyl]-Ala-Ala-Phe-p-aminobenzoate, a specific active-site-directed inhibitor of endopeptidase-24.15, indicating that this enzyme was responsible for the observed interconversions. The data indicate that synaptosomal membranes contain enzymes that can potentially generate and degrade both leucine- and methionine-enkephalin.

ACTH neuromodulation of the developing motor system and neonatal learning in the rat.

ACTH peptides influence the developing nervous system during the first three weeks of life in the rat. ACTH 4-10 and Org 2766 (10.0 micrograms/kg) accelerate the expression of motor hyperactivity usually exhibited in 15-day old normal animals, with ACTH 4-10 increasing the force of extensor digitorum longus muscle contraction amplitude. Following cold stress and peptide treatment, rate changes in motor activity from one age to the next are dramatically enhanced, with vertical activity being exhibited at an earlier age than controls. Grasping ability is similarly enhanced in 13-day old ACTH 4-10-treated animals. The retention of a T-maze learning paradigm is significantly enhanced in 16-day old ACTH 4-10 (10.0 micrograms/kg)-treated and Org 2766 (0.01 micrograms/kg)-treated animals, with these animals running the maze significantly faster than controls. Peptide treatment appears to reverse the apparent turning preference in the maze during extinction. It is suggested that ACTH peptides modulate the organization of the nervous system and facilitate neurotransmission, and may act on dopaminergic and cholinergic neurotransmitter systems. Motor behavior seems to reflect underlying neural substrates that are integrated to produce the overt behavior of the organism.

Development and regeneration of motor systems under the influence of ACTH peptides.

ACTH peptides exert quantitative and qualitative influences on the formation and maturation of motor units in developing and regenerating neuromuscular systems. ACTH 4-10, administered daily (10 micrograms/kg. s.c.) from the day of birth, accelerated the rate at which muscle strength developed in the immature rat, the effect of this peptide being most marked in animals 11-15 days old. A similar increase in grasping time occurred in ACTH 4-10 treated animals, indicating that the peptide affects neuronal maturation at a time in development when organization and maturation of the neuromuscular system is most active. The synthetic analogue of ACTH 4-9 (Org 2766), administered in the same dosage, had little effect on these parameters, indicating a differential sensitivity to these similar peptides. Elevated circulating titers of ACTH, whether exogenous (0.2 U ACTH 1-39 IP daily), or endogenous (adrenalectomy), stimulated the formation of more functional motor units, as indicated by increased amplitude of muscle action potentials and tetanic tension following nerve stimulation. ACTH appears to favor the recovery of high threshold, small-size motor units. Fine control of muscle function in peptide-treated animals is partially restored, as indicated by the return of stepwise recruitment to an extent not seen in the reinnervated, saline-treated controls.

Peptide influences on the development and regeneration of motor performance.

ACTH 1-39 (0.2 U IP daily for up to 18 days) has a beneficial effect on the functional reorganization of regenerating motor units of the extensor digitorum longus (EDL) in the adrenalectomized adult rat following crushing of the peroneal nerve. Motor unit activity (maximum twitch tension amplitude/mean increment in twitch tension as voltage is increased by 0.1 V gradations) and nerve-muscle efficiency (tetanic tension from indirect stimulation/tetanic tension from direct stimulation of EDL) were enhanced by ACTH 1-39. Other electrophysiological and contractile parameters were unaffected by the peptide. Spontaneous motor activity in cold stressed 13 day old rats was prolonged by Org 2766, a substituted analogue of ACTH/MSH 4-9, (0.1 microgram/kg daily) but unaffected by the same dosage of ACTH/MSH 4-10. The responsiveness of developing and regenerating motor systems to neuropeptides indicates a plasticity of neuronal connections, which depends on peptide sequence, dosage and the physiological state of the animal (normal, depressed, regenerating or developing, at rest or stressed).