Effects of 6 months of endurance training on neutrophil functions to produce reactive oxygen species and mental states in male long-distance runners.

To clarify changes of neutrophil functions, mental conditions and relationships among them, 19 male elite long-distance runners participated in this study for 6 months. Examinations, with informed consent, were carried out once a month. According to the results of physical characteristics, it was thought that training intensity was reduced after the main race, Hakone-Ekiden. Neutrophil functions were estimated by indices of reactive oxygen species production, determined by luminol- and lucigenin-dependent chemiluminescence (LmCL and LgCL, respectively) and cytochrome c reduction methods. The peak times (PT) in LmCL and LgCL (LgPT) were most prolonged in January and December, respectively. The peak heights (PH) in LmCL (LmPH) were enhanced in February. Decreased levels of negative categories in the profile of mood state (POMS) questionnaire and the total mood state (TMS) of POMS were observed in February without significance. Correlation analysis using measured values revealed significant negative correlation between LmPH and negative categories in POMS; however, these correlations were possibly a mere appearance, caused by personal differences. After eliminating personal differences, LgPT correlated positively to depression (p< 0.05), anger (p< 0.05), fatigue (p < 0.01) and TMS (p< 0.05). These results suggest that the mean time from the recognition of foreign matter to the maximum production of superoxide from neutrophils is prolonged in the mentally suppressed conditions found under continuous physical training.

Assessment of training effects on autonomic modulation of the cardiovascular system in mature rats using power spectral analysis of heart rate variability.

OBJECTIVE: To clarify the effects of forced or voluntary exercise on autonomic modulation of the cardiovascular system, we monitored changes in autonomic nervous activity in a mature rat by spectral analysis of the heart rate (HR) during a 10-week training period. METHODS: Male Wistar rats implanted with a radio-telemetry system were divided into three groups at 18 weeks of age: (1) Control group (n = 8); (2) Voluntary group (n = 6), which were housed separately in a cage with a running wheel; (3) Forced group (n = 6), which were exercised on a treadmill (35 m/min, 15 min/day, 5 days/week). The electrocardiogram was analyzed by the maximum entropy method into two main oscillations, low-frequency (LF) and high-frequency (HF) oscillations, respectively. LF and HF are considered to be markers of both sympathetic and parasympathetic modulations and parasympathetic modulation, respectively. RESULTS: Average running distances of the Voluntary group were more than twofold higher than those of the Forced group. HR levels in the Forced group were lower than those in the Control group. LF and HF levels in the Control and the Forced groups were almost the same during the experiment, and those in the Voluntary group showed a tendency to decrease. CONCLUSION: The results in the Voluntary and the Forced groups suggest that cardiovascular adjustments are not simply caused by the quantity of exercise. In the Voluntary group, both sympathetic and parasympathetic activity may decrease with a predominance of sympathetic activity. Conversely, in the Forced group, the baroreflex may be hyper-activated by the undesired treadmill running and handling stress.

Effects of summer training camp on serum opsonic activity and plasma cytokine levels in female long-distance runners.

In a summer training camp, strenuous physical exercises are carried out to improve endurance ability. The effects of the training stress on the immune system have been studied extensively. However, less attention has been paid to non-specific immunological changes. Serum opsonic activity (SOA) is a more direct and suitable indicator of non-specific humoral immunity. In this study, we used the luminal-dependent chemiluminescence (LmCL) to measure reactive oxygen species generation from pooled human neutrophils as an indicator of SOA. We also measured plasma myeloperoxidase (MPO) and plasma cytokine levels. Twenty-two female college runners took part in this study after giving their written informed consent. There was no significant difference in the physical characteristics and serum enzyme levels. However, significant differences were observed in the changes of blood property and plasma cytokine levels. According to MPO levels, neutrophils in vivo may be deactivated during the camp. Positive correlations between fluctuations of pro- and anti-inflammatory cytokine levels were observed in this study. According to the results of peak time and peak height of LmCL, SOA seems to increase at first qualitatively and then quantitatively during the summer camp. The increased SOA level may compensate for the decrease in neutrophil activity.

Effects of a 21 day metabolic study on serum opsonic activity in female college students, assessed by a chemiluminescence technique.

A metabolic study to evaluate nutritional balance, in which subjects have to eat all the foods supplied and maintain a set schedule, is thought to be stressful for the subjects. The effects of stress on the immune system have been studied extensively. However, there are no studies of non-specific immunological changes during a metabolic study. Serum opsonic activity (SOA) is a direct and suitable indicator of non-specific humoral immunity. In this study, we used luminol-dependent chemiluminescence (LmCL) to measure reactive oxygen species (ROS) generation from pooled human neutrophils as an indicator of SOA. We also measured serum immunoglobulin levels and plasma myeloperoxidase (MPO) activity. Eleven female college students took part in this 21 day metabolic study after giving their written informed consent. The results obtained suggest that the metabolic study has almost no effect on immunoglobulin levels. According to MPO levels, neutrophils in vivo may be deactivated to some minor extent. In contrast to these results, peak time (PT) and peak height (PH) of LmCL were changed significantly during the metabolic study. In conclusion, SOA increased during the 21 day metabolic study. There was no significant correlation between SOA and serum immunoglobulin levels on any of the study days.

Comparison of effects of in vivo nitrogen dioxide exposure starting from different periods on alveolar macrophage activity, assessed by a chemiluminescence technique in Brown-Norway rats.

Nitrogen dioxide (NO2) has been extensively studied for its immune modulating effects on pulmonary cells. Alveolar macrophages (AMs) play an important role in pulmonary immunity. The Brown-Norway (BN) rat has been studied as a high-risk model of allergic diseases. In this study, BN rats were exposed to NO2 from the embryonic or weanling period (EP or WP, respectively). To evaluate the effects of NO2 exposure on pulmonary immunity, the activity levels of rat AMs were assessed as reactive oxygen species-generating capacity, measured by a chemiluminescence (CL) technique, and as cytokine-producing ability. Except for 0.2 ppm of NO2 exposure, the CL responses of AMs obtained from the WP group at 12 weeks old were suppressed significantly. Changes of the cytokine-producing levels suggest that inflammatory reactions are terminated at 12 weeks in the EP group. Correlations between the CL responses and the cytokine levels reveal that NO2 exposure may modulate the direction of AM activation. The CL technique is thought to be useful to evaluate changes in AM activity. In this study, the results suggest that, using the high-risk model of allergic diseases, NO2 exposure from the weanling period has stronger effects on AM activity.

Bowel habits before and during menses in Japanese women of climacteric age: a population based study.

A good deal of data are available on the bowel habits of pre-climacteric females during menstruation. Few studies have examined the same subject in females of climacteric age but who are still menstruating, so the present study was undertaken to examine the bowel habits in menstruating females in this age group. Subjects (n = 246) were residents of a city in northern Japan, aged from 45 to 55 years old and who were still menstruating. Their every-day state of defecation and fecal characteristics were assessed with regard to four parameters: bowel movement frequency, defecation state, fecal appearance and fecal consistency. Based on a perfect match to these four criteria, the subjects were assigned to two groups, the normal group and the constipation group, and changes were assessed by enquiring how their bowel habits differed immediately before and during menses compared with their usual state. Immediately before menstruation, in the constipation group feces became harder and looser in 22.1% and 13.7%, respectively, compared with 8.7% and 9.5% in the normal group. On the other hand, during menstruation in the constipation group, feces became harder and looser in 11.6% and 16.8%, respectively, compared with 5.7% and 8.9% in the normal group. In other words, the constipation group showed a greater change immediately before and during menstruation compared with the normal group, though the differences were not significant. Our data thus suggest that the changes in the bowel habits of women of climacteric age at menstruation are greater for those suffering from constipation than those who are not.

A tube-fed liquid formula diet containing dietary fiber increased stool weight in bed-ridden elderly patients.

OBJECTIVE: The number of bedridden elderly patients who need a liquid diet has been increasing. We evaluated the usefulness of a tube-fed liquid diet containing cellulose and oligosaccharide for elderly bedridden patients. METHODS: Twenty-two elderly bedridden patients were given a liquid diet containing cellulose and lafinose for 4 wk. Parameters associated with blood and stool weight were measured during this period. RESULTS: Stool weight increased by 32.0 g in men and 8.8 g in women at 2 wk and remained at these weights up to 4 wk (P < 0.05 before and after 2 wk, 3 wk, and 4 wk in men, and P < 0.05 before and after 3 wk in women). Body weight increased in men (0.5 kg) and women (0.7 kg) 15 d after starting the diet and then remained at these values up to 29 d thereafter, and statistically significant differences were not seen. There were no changes in nutritional parameters such as total protein. CONCLUSION: No significant changes were observed in the general condition of elderly patients, with the exception of a positive increase in fecal weight, by 30-d supplementation of a liquid diet containing cellulose and lafinose.

A competitive marathon race decreases neutrophil functions in athletes.

A full marathon is the longest running race in official track events and is a form of acute exercise. However, no studies have examined the acute neutrophil function response to a competitive marathon race. Thirty-six male athletes who had just completed the 42.195 km course of the 50th Beppu-Oita Mainichi Marathon were enrolled in this study. Neutrophil oxidative burst activity, phagocytic activity and expression of CD11b and CD16 per cell were measured by flow cytometry immediately before and after the marathon. Total leukocyte/neutrophil counts increased significantly (p < 0.001), whereas total oxidative burst activity per neutrophil cell decreased significantly after the race (p < 0.001). Furthermore, total phagocytic activity per neutrophil cell also decreased after the race, although it was not significant (p = 0.08). Although CD11b expression per cell did not change, the expression of CD16 per cell significantly decreased (p < 0.001) after the race. In conclusion, a competitive marathon race decreased neutrophil functions (oxidative burst activity and phagocytic activity), which may be partly due to a decrease in CD16 expression. The increase in total neutrophil counts might reflect a compensatory response to counteract the decrease in neutrophil functions.

Impact of a competitive marathon race on systemic cytokine and neutrophil responses.

PURPOSE: To investigate whether cytokines and neutrophils mediate exercise-related pathogenesis, we examined their responses and possible association after exhaustive exercise. METHODS: Plasma and urine samples were obtained from 10 male runners before and after a 42.195-km marathon race. Major cytokines and neutrophil activation markers [myeloperoxidase (MPO) and lactoferrin (LTF)] were measured by enzyme-linked immunosorbent assays. Functional modulation of standard neutrophils and monocytes by plasma was determined on their luminol-dependent chemiluminescence responses. RESULTS: The race induced peripheral neutrophilia accompanied by an increase in band neutrophils and monocytosis. Plasma MPO and LTF concentrations increased significantly by 1.8 and 1.4 times after the race. There was a greater increase in urine concentrations of MPO and LTF, 12.3 and 3.5 times after exercise, respectively, suggesting that neutrophil activation occurred and that renal clearance exceeded the increase in plasma concentrations. Plasma interleukin (IL)-6, IL-8, IL-10, granulocyte colony-stimulating factor (G-CSF), macrophage CSF (M-CSF), and monocyte chemotactic protein 1 (MCP-1) increased significantly after the race, and urine IL-1beta, IL-6, G-CSF, M-CSF, and MCP-1 increased significantly. The plasma IL-6 responses correlated with the increases of band neutrophil count (r = 0.860, P < 0.01), suggesting IL-6-mediated bone marrow release of neutrophils. Furthermore, the increases in urine MPO concentration were correlated with increases in urine IL-6 (r = 0.868, P < 0.01) and G-CSF (r = 0.875, P < 0.01), suggesting that these cytokines promoted neutrophil activation. However, preincubation of neutrophils and monocytes with postexercise plasma could not cause priming responses, possibly because of the exercise-induced enhancement of plasma antioxidant activity. CONCLUSION: Although many cytokines recruiting and priming neutrophils and monocytes were secreted and functional after exhaustive exercise, overwhelming antioxidant and antiinflammatory defenses were induced, preventing exercise-induced oxidative stress.

Assessment of training effects on activity levels of alveolar macrophage in matured rats using chemiluminescent technique.

Chemiluminescence responses have been used for the evaluation of phagocyte function. In this study, to evaluate effects of training started after maturation on pulmonary immunity, the activity levels of rat alveolar macrophages (AMs) were assessed as reactive oxygen species (ROS) generating capacity, measured by lucigenin- and luminol-dependent chemiluminescence, using a parallel luminometer. One group of male Wistar rats started training at 11 weeks old and another group at 17 weeks old. The experimental period was 12 weeks, and about half of the rats were sacrificed after 6 weeks. The forced and voluntary exercises affect the mean levels of body weights and cell populations in the bronchoalveolar lavage fluid in younger animals; however, the voluntary exercise group in younger animals seemed to adapt after 12 weeks. By contrast, chemiluminescence responses in older rats observed after 6 weeks suggest that AMs are primed, and the maximum releasing activities of ROS are reduced. These changes in AM activity may be caused by the exercise and separation stresses and the rats may adapt to those stressors after 12 weeks. The chemiluminescent technique is thought to be useful to evaluate the changes of AM activity.