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Glutamine is an important amino acid for immune function. Though high intensity and prolonged exercise decreases plasma glutamine concentration and causes immune suppression, the relationship between neutrophil functions and glutamine has not yet been found. The purpose of this study was to investigate the impacts of glutamine supplementation on neutrophil function. Twenty-six male university judoists were recruited. Subjects were classified into glutamine and control groups. The glutamine group ingested 3000 mg of glutamine per day and the control group ingested placebo for 2 weeks. Examinations were performed at the start of preunified loading exercise (pre-ULE), then 1 and 2 weeks after ULE (post-ULE). Reactive oxygen species (ROS) production, phagocytic activity, serum opsonic activity and serum myogenic enzymes were measured. Differences between the levels obtained in pre-ULE and post-ULE for the two groups were compared. In the glutamine group, ROS production activity increased 1 week after ULE, whereas it was not observed in the control group (P < 0.001). Though myogenic enzymes increased significantly after ULE (P < 0.001), the glutamine group remained unchanged by supplementation during ULE. Glutamine supplementation has prevented excessive muscle damage and suppression of neutrophil function, especially in ROS production activity, even during an intensive training period., (Copyright © 2013 John Wiley & Sons, Ltd.)

The concept of peaking ensures that athletes have trained to attain their absolute peak performance levels prior to a competition. This study investigates the effects of peaking on the functions of neutrophils and lymphocytes in university soccer players during a five-day soccer training camp followed by two weeks of tapering. The study subjects were 22 soccer players who were members of a university soccer club. We carried out our investigation during a five-day training camp and two weeks after the training camp (i.e. the tapering period). We measured body composition, immune-related parameters (leukocyte count, neutrophil count, lymphocyte count, immunoglobulins and complements), myogenic enzymes, superoxide dismutase activity (SOD) and neutrophil functions [reactive oxygen species (ROS) production, phagocytic activity, serum opsonic activity and lymphocyte subtypes]. Leukocyte and neutrophil counts tend to increase after the training camp compared with values before the training camp, and recovered during the conditioning period, although the final values were still lower than those before the training camp. The amount of ROS production per neutrophil and level of SOD decreased significantly during the conditioning period compared with before the training camp (p < 0.05, p < 0.01, respectively). Levels of Th1 cells decreased significantly during the conditioning period compared with the training camp (p < 0.05). In conclusion, the period of two weeks for the tapering period was considered insufficient to allow the athletes' immune function to recover completely after an intensive training camp., (Copyright © 2013 John Wiley & Sons, Ltd.)

We assessed the association of neutrophil function with glycated hemoglobin (HbA1c) levels in a Japanese general population. Participants were 809 males and females who were over 20 years old living in the Iwaki region in Aomori Prefecture located in northern Japan. Lifestyle parameters (smoking, alcohol consumption, and exercise habits), HbA1c and neutrophil function such as reactive oxygen species (ROS) production capability and phagocytic activity (PA) were measured. ROS production capability was measured before and after phagocytic stimulus to obtain basal ROS production and stimulated ROS production. Level of HbA1c had a positive correlation with basal ROS production (p=0.053), a negative correlation with stimulated ROS production (p=0.072) and PA (p=0.059) only in post-menopausal groups, and not in pre-menopausal groups. However, there were no correlations between levels of HbA1c and neutrophil functions in male. In conclusion, in the present study, despite the presence of diabetes, chronic hyperglycemia was found to cause an increase in daily basal ROS production of neutrophils, and increased susceptibility to infection caused by reduced neutrophilic reaction in females in their menopause. Therefore, from the oxidative point of view, strict glycemic control is necessary to prevent post-menopausal females from developing diabetic complications in spite of the presence of diabetes., (Copyright © 2013 John Wiley & Sons, Ltd.)

In this study, changes in physical fatigue and biological functions of Japanese female soccer players were investigated by determining changes in neutrophil and lymphocyte functions. Study subjects included 18 female soccer players. Body composition, serum myogenic enzymes, neutrophil function, including reactive oxygen species (ROS) production capability, phagocytic activity (PA) and serum opsonic activity, as well as lymphocyte subpopulation were measured before and after a soccer match. Levels of myogenic enzymes (AST, ALT, CK and LDH) and immunoglobulins (IgG and IgA) and complements (C3) increased significantly after the match. In addition, leukocyte, neutrophils and lymphocyte counts increased whereas total PA decreased significantly. The number of T and Th1 cells (subsets of T helper cells) decreased whereas Th2 increased significantly. In addition, the number of B cells increased and NK cells decreased significantly after the match. The match was found to result in degenerative changes in and damage to athlete muscle tissues together with damage- and change-mediated stress. These data also suggest a post-match accelerated inflammatory reaction and potential immunosuppression as indicated by reductions in neutrophil PA and lymphocyte functions., (Copyright © 2012 John Wiley & Sons, Ltd.)

We examined changes in neutrophil function of soccer players over a 10-month period and determined its effectiveness as an index for athlete physical condition. Subjects included 21 male professional Japanese soccer players. Data on body composition, myogenic enzymes and neutrophil function were obtained before and after 2 h of training at 3 investigation points: one week before opening season, at season mid-point, and one week before the last game of the season. As a result, change ratios of myogenic enzyme levels before and after the 2-hr training session at the third investigation point were significantly higher compared to the two other points. Reactive oxygen species production and phagocytic activity significantly increased after 2-hr training session at point 1, although the extent of the increase became smaller over time and ROS production capability decreased significantly by point 3 assessment. Fatigue, especially muscle fatigue, chronically accumulated along with a gradual decrease in neutrophil immune function over the 10-month season. Therefore, determination of neutrophil function can be used as a useful index to assess and understand an athlete's physical condition., (Copyright © 2012 John Wiley & Sons, Ltd.)

We investigated the effects of dehydration after a judo practice session on player muscle and immune functions. Subjects included 25 female university judoists. Investigations were performed before and after 2.5 h of regular judo practice. Body composition, serum enzymes (myogenic enzymes, immunoglobulins and complements), neutrophils counts, reactive oxygen species (ROS) production capability, and phagocytic activity (PA) were measured. Subjects were divided into two groups according to level of dehydration after practice (mild dehydration and severe dehydration groups) and results were compared. Creatine kinase was found to increase significantly after practice. In addition, neutrophil count also increased significantly after practice in both groups. The changing ratios of IgA, IgG and C3 observed in the mild dehydration group were significantly higher than those in the severe dehydration group. In the severe dehydration group, post-practice PA/neutrophil had decreased significantly. Significant positive correlations were found between severity of dehydration and changing ratios of IgA, IgG, IgM, C3, C4 and ROS production capabilities, whereas no significant association was seen with PA and/or serum SOD activity. These results suggest that dehydration resulted in immunosuppression, including decreased neutrophil function., (Copyright © 2012 John Wiley & Sons, Ltd.)

We studied the effects of a high intensity and high frequency 3-month training program on muscle damage and neutrophil function in male judoists. The study included 15 male judoists who started intensive judo training program after a 6-month break. Creatine kinase (CK), neutrophil counts and reactive oxygen species (ROS) production capability as well as phagocytic activity (PA) of neutrophils were measured at 2 stages; entering university (pre-training) and after 3-month training (post-training). At both points, we investigated parameters three times: just before, immediately after and 24 h after a 2-h practice session. Practice-mediated change in CK was lower at post-training than at pre-training. Neutrophil count significantly increased after 2-h practice but recovered 24 h later whereas it showed no subsequent and further increased at 24 h post-practice. Although neutrophil ROS production capability and PA both decreased (breakdown) after practice session, ROS production capability increased and PA decreased (well-adapted) at the post-training. Long-term training strengthened muscular function and improved neutrophil reaction against practice-mediated stress., (Copyright © 2012 John Wiley & Sons, Ltd.)

We evaluated the relationships between neutrophil-related functions and serum selenium (Se) concentration in the general population. We examined 800 subjects who had participated in the Iwaki Health Promotion Project in 2005 to determine the relationships between serum Se concentration and neutrophil-related functions such as the production capability of reactive oxygen species (ROS), phagocytic activity, and serum opsonic activity (SOA). In nonstimulated neutrophils, i.e., in neutrophils at their baseline condition before the application of the phagocytic stimulus, the serum Se concentration tends to be high and the ROS production tends to be low. With regard to SOA, there was a significant negative correlation between lucigenin-dependent chemiluminescence and serum Se concentration in both men and women. Moreover, in women, a significant negative correlation was observed between luminol-dependent chemiluminescence and serum Se concentration. These results suggest that subjects with a lower serum Se concentration may be exposed to a greater chronic oxidative stress due to neutrophil ROS production. In addition, the findings of our study suggest that women rather than men benefit more from Se against oxidative stress.

In recent years, the effects of smoking and excessive alcohol consumption on immune function have been studied, due to a high prevalence of infection or cancer in heavy drinkers, and the combination of smoking and drinking was considered to be a carcinogenic risk. However, the effect of smoking and drinking on systemic immune function has yet to be clearly understood. In this study, we investigated neutrophil functions (reactive oxygen species (ROS) productive activity, phagocytic ability and serum opsonic activity) and their relationship with alcohol consumption or amount of smoking. In total there were 731 male and female adult subjects who participated in the Iwaki Health Promotion Project in 2005. Multiple regression analysis showed a trend of increased ROS production in male subjects and a statistically significant decrease was observed in phagocytic activity caused by smoking in female subjects. In other words, oxidative stress caused by smoking in male subjects may be involved in ROS production from neutrophils. Decreased phagocytic activity of neutrophils caused by smoking suggests that host defense functions were impaired in female subjects. A relationship between neutrophil functions and the amount of alcohol consumption was not observed., (Copyright © 2011 John Wiley & Sons, Ltd.)

The aim of this review is to provide a summary of the known effects of exercise on neutrophil immune functions of athletes. We measured three neutrophil immune functions (i.e., phagocytic activity (PA), reactive oxygen species (ROS) production, and serum opsonic activity (SOA)) in various types of exercise. The following is our recent findings. (1) A regular exercise increases ROS production and decreases PA. We call this change a normal pattern, and an abnormal pattern except this change. (2) A prolonged, strenuous activity (e.g., rugby match and marathon) decreases both ROS production and PA. This is one of the abnormal pattern. (3) The exercise loading performed after a camp training decreases ROS production whereas PA does not change. This is another abnormal pattern. (4) When judoists who had stopped judo training for 6 months restarted their training, the exercise loading at the beginning of their training decreases PA whereas ROS production does not change. This is another abnormal pattern. (5) A regular exercise 2 months after the beginning of their training increases ROS production and decreases PA. This change is a normal pattern. SOA showed a similar pattern of changes to ROS under all conditions. The changes in neutrophil immune functions after performing various exercises might result from the balance between external factors (intensity and style of exercise) and internal factors (e.g., fatigue and physical pain). Therefore, the changes in three neutrophil immune functions after exercise might be an index of athletes' condition.

We have evaluated the relationship between exhaled carbon monoxide (CO) level and neutrophil-related functions such as reactive oxygen species (ROS) production capability, phagocytic activity and serum opsonic activity in the general population. Serum opsonic activity was determined by measuring the effects of serum on neutrophil ROS production capability using lucigenin- and luminol-dependent chemiluminescence (LgCL, LmCL). LgCL is associated with the detection of O(2)(-) , whereas LmCL mainly detects H(2)O(2) and HOCl, which are higher reactive oxygen radicals. In females, exhaled CO level was found to have positive associations with ROS production capability and LgCL. However, the opposite tendency was seen between exhaled CO level and LmCL in both genders. This result suggests that neutrophil ROS production in females may have contributed to oxidative stress, which led to the increases in intrinsic CO and exhaled CO consequently. Such changes then may have inhibited the process of changing reactive oxygen radicals into higher oxidizing potential levels., (Copyright © 2010 John Wiley & Sons, Ltd.)

In order to develop a predictive marker of overtraining in athletes, we examined the changes in neutrophil function [reactive oxygen species (ROS) production capability and phagocytic activity (PA)] for 10 male and 13 female judoists attending a training camp. Measurements were taken four times in total--immediately before and after a 2 h unified exercise loading (UEL) performed 1 day before (Pre-Camp) and the day after the 7 day training camp (Post-Camp). UEL-mediated aspartate aminotransferase was higher at Post-Camp than at Pre-Camp in females but not in males. Post-Camp leukocyte/neutrophil counts after the UEL significantly increased in females but not in males. The rate of change in C4 was significantly smaller in females than in males at Post-Camp. Only ROS significantly decreased without any compensation (increase in PA) being made at Post-Camp in females. In conclusion, this finding, namely that ROS significantly decreased only at Post-Camp without any compensatory mechanism (increase in PA), would suggest that the training camp imposed greater loading on females than males. This consideration was supported by the atypical aspartate aminotransferase, leukocyte/neutrophil counts and C4 findings which were seen at Post-Camp only in females. Therefore, regularly examining neutrophil functions such as ROS and PA might be a good preventative measure against overtraining in athletes participating in training camps.

Helicobacter pylori infection has been reported to cause enhanced reactive oxygen species in the gastric mucosa. We examined the relationship between H. pylori infection and neutrophil function of peripheral blood. The subjects were 904 volunteers who participated in the Iwaki Health Promotion Project in 2005. 158 subjects who were infected with H. pylori in 2005 also participated in this project in 2006 and were categorized into two groups: the eradication group, in which H. pylori was successfully eradicated during the 12 month period, and the non-eradication group, in which eradication was unsuccessful or the subjects did not receive eradication therapy. The laboratory assays performed were: a titre of H. pylori antibody; neutrophil counts; and oxidative burst activity (OBA) of neutrophils. Logistic regression analysis was executed, with H. pylori infection as the dependent variable and other items as the independent variables. OBA showed an inverse association with H. pylori infection in 2005. Additionally, when comparing the eradication and non-eradication groups, the change rates of OBA between 2005 and 2006 did not show any significant difference. It was concluded that H. pylori infection does not lower OBA, but those individuals in whom OBA was lower were more prone to H. pylori infection., ((c) 2008 John Wiley & Sons, Ltd.)

In order to examine in detail the influence on the neutrophil immune function in sumo wrestlers of performing traditional and original training we examined changes in the neutrophil immune function in 17 male amateur university sumo wrestlers (aged 20.2 +/- 1.5 years), before ('Pre') and after the training ('Post') for 2.5 h under fasting conditions. Assays included blood leukocyte and neutrophil counts, serum concentration of immunoglobulins, complements, myogenic enzymes and neutrophil oxidative burst activity (OBA) and phagocytic activity (PA). Myogenic enzymes, neutrophil counts, the ratio of neutrophil counts:leukocyte counts significantly increased and immunoglobulins and complements decreased in Post compared with Pre. There was a positive correlation between the change of neutrophil counts before and after the training and the change of creatine kinase (r = 0.667, p < 0.01). The Post OBA significantly increased and PA significantly decreased compared with Pre. It was concluded that sumo training causes muscular damage and an increase in the neutrophil count as a response. In this time, although OBA increased, PA decreased after training. Compensation between PA and reactive oxygen species production may exist to maintain the overall integrity of the neutrophil immune function., ((c) 2008 John Wiley & Sons, Ltd.)

Objectives: To clarify the effects of high-intensity and high-frequency long-term/chronic training on neutrophil function and serum levels of myogenic enzymes in male university judoists., Methods: The subjects were 24 male judoists who had stopped judo training for 6 months and then restarted their training. The following parameters were examined before and after a 2 h unified exercise loading (UEL) at the beginning of the restarted quotidian training (pre-training) and at 2 months, 4 months and 6 months thereafter: myogenic enzymes, neutrophil and leucocyte counts, and neutrophil phagocytic activity (PA) and oxidative burst activity as a measure of reactive oxygen species (ROS) production capability., Results: Myogenic enzymes that were measured after UEL at all four points significantly increased except for creatine kinase at the 2-month point (p<0.01 in each) and neutrophil counts significantly increased after UEL at the pre-training, 2-month and 4-month points (p<0.01 in each), but these changes became smaller from the 2-month point. PA significantly decreased after UEL at the pre-training and 2-month points (p<0.01 in each), but no change was seen at the 4-month and 6-month points. On the other hand, no change in ROS production per cell after UEL was seen at the pre-training point, but it significantly increased after UEL at the 2-month, 4-month and 6-month points (p<0.01 in each)., Conclusion: The changing rate of the levels of UEL-mediated myogenic enzymes, neutrophil mobilisation and neutrophil function was seen to decrease at the 2-month, 4-month and 6-month assessments, compared with the pre-training point: these may comprise at least some of the long-term training effects.

The present study examined the effects of judo training on neutrophil and related functions. We measured and studied changes in the neutrophil and its related functions in 22 male university judoists immediately before (Pre values) and immediately after (Post values) a 2 h training session: reactive oxygen species (ROS) production capability, phagocytic activities (PA) and serum opsonic activity (SOA). Neutrophil count in whole blood, myogenic enzymes (creatine kinase, lactate dehydrogenase, aspartate aminotransferase and alanine aminotransferase), immunoglobulins (IgG, IgA and IgM) and complements (C3 and C4) in serum were also measured. The Post values of the neutrophil count, myogenic enzymes and IgG increased significantly compared with the Pre values. ROS production capability and SOA also significantly increased following training, although PA showed a slight decrease (but not statistically significant). Taking the findings of our previous studies into consideration, three major neutrophil or related functions, namely ROS production capability, PA and SOA, might compensate for each other to maintain the overall integrity of the neutrophil immune function, in that ROS and SOA increased to compensate for the slight decrease in PA, or PA slightly decreased to compensate for the increase in ROA and SOA after exercise., ((c) 2008 John Wiley & Sons, Ltd.)

Objective: Athletes undergoing weight reduction are recognised as being more prone to infection. Few studies exist for athletes on the weight reduction-mediated changes in neutrophil function and related activities such as reactive oxygen species (ROS) production capability, phagocytic activity (PA) and serum opsonic activity (SOA)., Methods: 16 Japanese female university judoists were examined in the early morning of the first day (pre-values) and the last day (post-values) of a 20-day pre-competition training period. Of the 16 subjects, 8 needed to reduce weight (WR group) and the other 8 did not (control group). The parameters assessed were the neutrophil count, serum immunoglobulins and complements, myogenic enzymes, ROS production capability, PA and SOA., Results: Comparing the post-values with the pre-values, ROS production significantly increased in both groups (p<0.01 for both). PA significantly decreased in the WR group (p<0.05); it also decreased in the control group but the decrease was not significant. SOA significantly increased in the control group (p<0.05), but showed no significant change in the WR group., Conclusions: The changes in the WR group were probably a direct consequence of the weight-reduction regimen coupled with the exercise regimen, suggesting that neutrophil parameters (ROS production, PA and SOA) had tended to deviate from their typical compensatory changes to maintain immune system homoeostasis.

Aims: To evaluate the influences of the accumulative effect of two consecutive rugby sevens matches (Sevens) on aspects of human neutrophil-related non-specific immunity., Methods: In seven players participating in the Japan Sevens, neutrophil reactive oxygen species (ROS) production capability and phagocytic activity were measured using flow cytometry, and serum opsonic activity (SOA) was assessed by measuring neutrophil ROS using the peak height of lucigenin-dependent chemiluminescence before and after two consecutive matches., Results: ROS showed no change immediately after the first match, and had significantly (P<0.05) increased 4 h later, but showed a decrease after the second match. Phagocytic activity showed no change immediately after the first match, but had significantly (P<0.01) decreased 4 h later, and showed a further decrease after the second match, although it was not significant. SOA significantly (P<0.01) increased after the first match, and still maintained its high 4 h later, but decreased after the second match. ROS production capability, phagocytic activity and SOA significantly (P<0.01) decreased after the second match., Conclusions: When rugby players play two consecutive Sevens matches, the exercise loading is thought to be hard, similar to that experienced during a marathon race and intensive or long training in a training camp, although the expected changes were not seen after the first match. Differences between after the first and the second matches may be due to the "cumulative effect".

In order to clarify the relationship between exercise and neutrophil function, we measured three major neutrophil and neutrophil-related functions, viz. the reactive oxygen species (ROS) production capability and phagocytic activity (PA) of neutrophils and serum opsonic activity (SOA), simultaneously before and after a unified loading exercise under three different sets of conditions. Thirteen female collegiate judoists were examined with a unified exercise loading (2 h) immediately before and after a 64 day training period. Immediately thereafter, the athletes took part in a 6 day intensified training camp, following which the same exercise loading was repeated. Responses from circulating neutrophils were estimated by comparing the two sets of values obtained before and after the two instances of exercise loading. The parameters assessed included neutrophil count, SOA, PA and ROS production capability. ROS production increased after the exercise loading performed immediately before and after the 64 day training period just before the camp, (p < 0.01) but decreased following the exercise loading performed after the camp (p < 0.05). This suggested depressed bacteriocidal capability of the circulating neutrophils. PA decreased after the exercise loading sessions imposed prior to and after the 64 day training period (p < 0.01) but did not change in the loading session after the camp. No changes were seen in SOA produced with the loading exercise either before the 64 day exercise period or before the camp, but increased significantly following the post-camp session (p < 0.05). In conclusion, athletic training-induced changes in immune functional activities of neutrophils, such as ROS production and PA, and neutrophil-related factors, such as SOA, may compensate for each other to maintain the overall integrity of the neutrophil immune function., (Copyright (c) 2006 John Wiley & Sons, Ltd.)

The effects of long-term training on the production of reactive oxygen species (ROS) from neutrophils and serum opsonic activity (SOA) remain to date unknown. The aim of this study was to evaluate the effect of 6 months training on ROS production and SOA in judoists. Fifty-six judoists were enrolled this study. White blood cell counts, serum creatine kinase (CK), asparate aminotransferase (ASAT), alanine aminotransferase (ALAT), lactate dehydrogenase (LDH) and ROS production from neutrophils, and serum opsonic activity (SOA) using the lucigenin and luminol probes, were measured before and after daily judo exercise (2 h) in March and September. The subjects started their training from March after no exercise for three months, and continued it for 6 months (until September). In March, myogenic enzymes such as CK, ASAT, LDH and neutrophil counts increased and immunoglobulins, complements and SOA decreased after daily judo exercise. Such significant changes were not seen in September. On the other hand, ROS significantly increased after daily judo exercise in both March and September, with no significant difference in the rates of change. In conclusion, 6 month training minimized the changes in SOA as well as muscle enzymes, neutrophil counts, serum immunoglobulins and complements. This could be categorized as a long-term training effect. However, no such change was seen in ROS.

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., (Copyright 2003 John Wiley & Sons, Ltd.)

The pathogenic roles of reactive oxygen species (ROS) have been implicated in ulcerative colitis (UC). The aim of this study was to examine the effects of ecabet sodium on ROS produced by human neutrophils, particularly after being primed by bacterial lipopolysaccharides (LPS). Neutrophils were isolated from six healthy volunteers. Each well of a 96-well microplate received neutrophil suspension (1.0 x 10(5) cells) and the plates were incubated at 37 degrees C for 30 min with or without E. coli LPS (f.c. 0.001 ng/ micro L). Ecabet sodium (f.c. 0-5.0 mg/mL) was added before starting or after finishing the incubation. Neutrophils were stimulated by opsonized zymosan (OZ; 1.0 mg/mL) or calcium ionophore (A21837; 0.3 micro mol/L) and luminol-dependent chemiluminescence response was measured using a Lumi Box H-1000. Ecabet sodium attenuated ROS production at a concentration of 5.0 mg/mL (p < 0.05) in LPS-primed neutrophils. However, attenuating effects were not significantly different when ecabet sodium was added before or after the incubation with E. coli LPS. Ecabet sodium may have some attenuating effects on ROS produced by human neutrophils even after neutrophils are primed by bacterial LPS. These results may explain, in part, the therapeutic effects of ecabet sodium for UC., (Copyright 2003 John Wiley & Sons, Ltd.)

Intensive exercise training decreases neutrophil functions in athletes. However, no studies to date have investigated the effect of irregular-interval training, such as is associated with judo training programmes, on neutrophil functions. The purpose of this study was to examine such effects. Thirty-seven male college judoists participated in this study. Neutrophil oxidative burst activity, phagocytic activity and expression of CD11b and CD16 per cell were measured by fl ow cytometry before and after judo training. Total neutrophil counts increased significantly from 2.98 +/- 0.82 to 7.95 +/- 1.80 x 10(3)/ microL (p < 0.001). The proportion of neutrophils producing reactive oxygen species (ROS) was increased significantly (p < 0.001). On the other hand, the phagocytic activity decreased after training, as shown by a decrease in the amount of ingested opsonized zymosan per cell (p < 0.001), possibly as a compensatory effect for the increased numbers of ROS-producing neutrophils. Expression of CD11b and CD16 per cell decreased by 20% and 30%, respectively, after judo training. In conclusion, judo training induced a decrease in phagocytic activity through the lowered expression of CD11b and CD16 on the surface of neutrophils, and increased the oxidative burst activity of neutrophils., (Copyright 2003 John Wiley & Sons, Ltd.)

This study investigated the effects of short-term weight reduction on neutrophil functions in female judoists. Sixteen actively competing female judoists were divided into two groups. Eight who required weight reduction were defined as the weight reduction group, and the remaining eight were used as the control. Blood samples were taken before, immediately after and 8 days after the match. Phagocytic activity and oxidative burst activity of neutrophils were measured by flow cytometry. In the weight reduction group, the phagocytic activity per cell decreased significantly at the end of weight reduction compared with the control group. The rate of neutrophils producing reactive oxygen species and the oxidative burst activity per cell increased significantly at the end of weight reduction in both the control and the weight reduction groups. We concluded that weight reduction, consisting of both intense exercise and energy restriction, might possibly cause both an increase in oxidative burst activity and decrease in neutrophil phagocytic activity in female judoists. However, although exercise increased oxidative burst activity, it did not affect neutrophil phagocytic activity alone. Therefore, to avoid this problem, female judoists are recommended to keep their weight within those limits determined by their class, and which can be reduced by exercise., (Copyright 2003 John Wiley & Sons, Ltd.)

Objective: The aim of this study was to examine the effects of low-level laser therapy (LLLT) on production of reactive oxygen (ROS) species by human neutrophils., Background Data: LLLT is an effective therapeutic modality for inflammatory conditions., Materials and Methods: The laser device used was the infrared diode laser (GaAlAs), 830-nm continuous wave (150 mW/cm(2)). After irradiation, ROS production by neutrophils was measured using luminol-dependent chemiluminescence (LmCL) and expression of CD11b and CD16 on neutrophil surface was measured by flow cytometry., Results: The LmCL response of neutrophils was reduced by laser irradiation at 60 min prior to the stimulation with opsonized zymosan and calcium ionophore. The attenuating effect of LLLT was larger in neutrophils of smokers than non-smokers, while the amount of produced ROS was larger in neutrophils of smokers. Expression of CD11b and CD16 on neutrophil surface was not affected by LLLT., Conclusion: Attenuation of ROS production by neutrophils may play a role in the effects of LLLT in the treatment of inflammatory tissues. There is a possible usage of LLLT to improve wound healing in smokers.

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.

Background: Short-chain fatty acids enema has been shown to be effective in the treatment of ulcerative colitis (UC). However, the mechanisms that lead to this response have not been well characterized. The aims of this study were to investigate the effect sodium butyrate has on reactive oxygen species (ROS) generation by human neutrophils, which are responsible for mucosal injury., Methods: Human neutrophils incubated with or without sodium butyrate were stimulated with opsonized zymosan (OZ) or phorbol myristate acetate (PMA). ROS generation was largely differentiated with flow cytometry assays of hydroethidine oxidation and dichlorofluorescein oxidation for superoxide anion and hydrogen peroxide respectively, and luminol-dependent chemiluminescence for myeloperoxidase-mediated oxidants., Results: Sodium butyrate (up to 50 mM) did not alter hydroethidine oxidation upon stimulation of the OZ or PMA. However, sodium butyrate at a concentration of 25 mM elevated dichlorofluorescein oxidation to 125 + 8% (P = 0.028) of control upon stimulation of OZ and to 191 +/- 30% (P = 0.0016) upon stimulation of PMA. Contrary to these results, sodium butyrate greatly inhibited chemiluminescence responses in a dose-dependent manner. The inhibition by 50 mM sodium butyrate was 61 +/- 6% upon OZ and 71 +/- 9% upon PMA, respectively., Conclusions: These data indicate that sodium butyrate up-regulates hydrogen peroxide generation but down-regulates generation of myeloperoxidase-mediated oxidants, the latter being more potent in killing microorganisms and in inducing tissue injury. A possible mechanism is suggested whereby sodium butyrate may inhibit myeloperoxidase activity and hence attenuate the destructive activities of neutrophils in UC.

Purpose: To investigate the effects of weight reduction as the result of exercise training and energy restriction on neutrophil function., Methods: Eighteen male competitive college judoists participated in the study. In a whole blood assay, oxidative burst activity, phagocytic activity, expressions of Fc gamma receptor 3 (CD16), and complement receptor 3 (CD11b) of neutrophils were measured on a per cell basis by flow cytometry at day 20, 5, and 1 before and at day 7 after the competition., Results: The rate of neutrophil producing reactive oxygen species decreased before the competition, whereas the oxidative burst activity per cell increased significantly in all subjects, which resulted in a significant increase of the total oxidative burst activity. However, there were no significant effect of energy restriction on oxidative burst activity. The rate of neutrophils incorporating opsonized zymosan decreased significantly with energy restriction. The total phagocytic activity of 10,000 neutrophils and the phagocytic activity per cell also decreased significantly by severe energy restriction. The surface antigen expressions of CD11b and CD16 were unaffected by weight reduction., Conclusions: The results suggest that with respect to the management of health conditions, weight reduction for judoists should be composed of exercise training and energy restriction should be moderate.

In order to examine that the effects of interferon alpha (IFN-alpha) on the production of reactive oxygen species from human neutrophils, we investigated in vitro effects of IFN-alpha on neutrophil chemiluminescence responses. Human neutrophils were incubated with 0-1 x 10(5) IU/mL IFN-alpha for 10-60 min just before stimulation with opsonized zymosan and phorbol 12-myristate 13-acetate. Luminol-dependent (LmCL) and lucigenin-dependent (LgCL) chemiluminescence responses were measured using a Lumi Box H-1000. The LmCL response was inhibited by pre-incubation with high concentrations of IFN-alpha (more than 1 x 10(4) IU/mL). In contrast, LgCL response was increased when neutrophils were pre-incubated with high concentration of IFN-alpha. Effects of pre-incubation of IFN-alpha were similar between CL responses stimulated with OZ and PMA. The increase of LgCL might be due to the inhibitory effect of IFN-alpha on myeloperoxidase degranulation, in view of the decreased LmCL responses., (Copyright 2001 John Wiley & Sons, Ltd.)

Decaglycerol monooleate (DGMO), a type of polyglycerol esters of fatty acids (PGEF), was evaluated for its in vitro effect on phagocytosis and respiratory burst activity of isolated human neutrophils using flow cytometric assay. Opsonized zymosan particles labelled with FITC (FITC-OZ) were employed as an indicator of phagocytosis. Fluorescence of FITC-OZ attached on to the surface of neutrophils was quenched by addition of trypan blue solution. After 10 minutes of incubation with DGMO up to a concentration of 10 mg/ml, neutrophil phagocytosis was not affected markedly. At the same time, the DGMO emulsion left little influence on complement receptor type three (CR3) that is associated with phagocytosis. On the other hand, oxidation of hydroethidine, which was used as an indicator of intracellular generation of reactive oxygen species (mainly for superoxide anion), was significantly inhibited by DGMO over 1 mg/ml. However, this phenomenon was not seen in DGMO-treated neutrophils when DGMO was removed after incubation. The present data suggest that DGMO does not affect phagocytosis of human neutrophils but down-regulates respiratory burst activity.

Decaglycerol mono-oleate (DGMO), a type of food additive (a polyglycerol ester of fatty acids), was evaluated for its in vitro effect on reactive oxygen species (ROS) generation by human neutrophils using a luminol-dependent chemiluminescence assay. Neutrophils were isolated by the Histopaque-1077/1119 double density gradient technique. Opsonized zymosan (OZ) and phorbol 12-myristate 13-acetate (PMA) were employed as artificial stimuli. DGMO inhibited the chemiluminescence response of OZ-stimulated neutrophils by 58% (p < 0.01) at a concentration of 0.5 mg/mL. DGMO inhibited the PMA-induced chemiluminescence by 40% (p < 0.01) at a concentration of 0.01 mg/mL. The inhibition phenomena could not be attributed to light absorption by DGMO alone, because the percentage chemiluminescence in the presence of DGMO is lower than the percentage transmittance of DGMO at 425 nm. These results suggest that DGMO downregulates ROS generation by human neutrophils., (Copyright 1999 John Wiley & Sons, Ltd.)