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David C. Nieman, Courtney L. Capps, Christopher R. Capps, Zack L. Shue, and Jennifer E. McBride

washout period and the second 4-week supplementation period), with runners following the exact same schedule (including time of day and day of the week). Plasma Cytokines Plasma was collected from ethylenediaminetetraacetic acid-treated blood, flash-frozen in liquid nitrogen, and stored at −80°C before

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Andrea T. Duran, Erik Gertz, Daniel A. Judelson, Andrea M. Haqq, Susan J. Clark, Kavin W. Tsang, and Daniela Rubin

Prader-Willi Syndrome (PWS), the best characterized form of syndromic obesity, presents with abnormally high fat mass. In children, obesity presents with low-grade systemic inflammation. This study evaluated if PWS and/or nonsyndromic obesity affected cytokine responses to intermittent aerobic exercise in children. Eleven children with PWS (11 ± 2 y, 45.4 ± 9.5% body fat), 12 children with obesity (OB) (9 ± 1 y, 39.9 ± 6.8% body fat), and 12 lean (LN) children (9 ± 1 y, 17.5 ± 4.6% body fat) participated. Children completed 10 2-min cycling bouts of vigorous intensity, separated by 1-min rest. Blood samples were collected preexercise (PRE), immediately postexercise (IP), and 15, 30, and 60 min into recovery to analyze possible changes in cytokines. In all groups, IL-6 and IL-8 concentrations were greater during recovery compared with PRE. PWS and OB exhibited higher IL-6 area under the curve (AUC) than LN (p < .01 for both). PWS demonstrated higher IL-8 AUC than LN (p < .04). IL-10, TNF-α, and IFN-γ did not change with exercise (p > .05 for all). Results indicate that children with PWS respond with increased Il-6 and IL-8 concentrations to acute exercise similarly to controls. Excess adiposity and epigenetic modifications may explain the greater integrated IL-6 and IL-8 responses in PWS compared with controls.

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Nicolette C. Bishop, Neil P. Walsh, Donna L. Haines, Emily E. Richards, and Michael Gleeson

Ingesting carbohydrate (CHO) beverages during heavy exercise is associated with smaller changes in the plasma concentrations of several cytokines. The influence of dietary CHO availability on these responses has not been determined. Therefore, the present study investigated the influence of pre-exercise CHO status on plasma interleukin (IL)-6, IL-10, and IL-1 receptor antagonist (IL-1ra) responses to prolonged cycling. Seven trained male cyclists performed a glycogen-lowering bout of cycling and were randomly assigned to follow a diet ensuring either greater than 70% (HIGH) or less than 10% (LOW) of daily energy intake from CHO for the next 3 days. On day 4 subjects performed an exercise test that comprised cycling for 1 hour at 60% Wmax immediately followed by a time-trial (TT) ensuring an energy expenditure equivalent to cycling for 30 min at 80% Wmax. Subjects repeated the protocol after 7 days, this time following the second diet. The order of the trials was counterbalanced. At 1 and 2 hours post-TT, plasma concentrations of IL-6 and IL-10 were 2-fold greater on the LOW trial than on the HIGH trial, and peak plasma concentrations of TL-1ra were 9-fold greater on the LOW trial than on the HIGH trial. These findings suggest that pre-exercise CHO status can influence the plasma cytokine response to prolonged cycling.

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David C. Nieman, Dru A. Henson, Steven R. McAnulty, Fuxia Jin, and Kendra R. Maxwell

The purpose of this study was to test the influence of 2.4 g/d fish oil n-3 polyunsaturated fatty acids (n-3 PUFA) over 6 wk on exercise performance, inflammation, and immune measures in 23 trained cyclists before and after a 3-d period of intense exercise. Participants were randomized to n-3 PUFA (n = 11; 2,000 mg eicosapentaenoic acid [EPA], 400 mg docosahexaenoic acid [DHA]) or placebo (n = 12) groups. They ingested supplements under double-blind methods for 6 wk before and during a 3-d period in which they cycled for 3 hr/d at ~57% Wmax with 10-km time trials inserted during the final 15 min of each 3-hr bout. Blood and saliva samples were collected before and after the 6-wk supplementation period, immediately after the 3-hr exercise bout on the third day, and 14 hr postexercise and analyzed for various immune-function and inflammation parameters. Supplementation with n-3 PUFA resulted in a significant increase in plasma EPA and DHA but had no effect on 10-km time-trial performance; preexercise outcome measures; exercise-induced increases in plasma cytokines, myeloperoxidase, blood total leukocytes, serum C-reactive protein, and creatine kinase; or the decrease in the salivary IgA:protein ratio. In conclusion, 6 wk supplementation with a large daily dose of n-3 PUFAs increased plasma EPA and DHA but had no effect on exercise performance or in countering measures of inflammation and immunity before or after a 3-d period of 9 hr of heavy exertion.

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Jeffrey B. Driban, Easwaran Balasubramanian, Mamta Amin, Michael R. Sitler, Marvin C. Ziskin, and Mary F. Barbe


Joint trauma is a risk factor for osteoarthritis (OA), which is becoming an increasingly important orthopedic concern for athletes and nonathletes alike. For advances in OA prevention, diagnosis, and treatment to occur, a greater understanding of the biochemical environment of the affected joint is needed.


To demonstrate the potential of a biochemical technique to enhance our understanding of and diagnostic capabilities for osteoarthritis.




Outpatient orthopedic practice.


8 subjects: 4 OA-knee participants (65 ± 6 y of age) and 4 normal-knee participants (54 ± 10 y) with no history of knee OA based on bilateral standing radiographs.


The independent variable was group (OA knee, normal knee).

Main Outcome Measures:

16 knee synovial-protein concentrations categorized as follows: 4 as pro-inflammatory, or catabolic, cytokines; 5 as anti-inflammatory, or protective, cytokines; 3 as catabolic enzymes; 2 as tissue inhibitors of metalloproteinases [TIMPs]; and 2 as adipokines.


Two anti-inflammatory cytokines (interleukin [IL]-13 and osteoprotegerin) and a pro-inflammatory cytokine (IL-1β) were significantly lower in the OA knees. Two catabolic enzymes (matrix metalloproteinase [MMP]-2 and MMP-3) were significantly elevated in OA knees. TIMP-2, an inhibitor of MMPs, was significantly elevated in OA knees.


Six of the 16 synovial-fluid proteins were significantly different between OA knees and normal knees in this study. Future research using a similar multiplex ELISA approach or other proteomic techniques may enable researchers and clinicians to develop more accurate biochemical profiles of synovial fluid to help diagnose OA, identify subsets of OA or individual characteristics, guide clinical decisions, and identify patients at risk for OA after knee injury.

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David C. Nieman, Francesca Ferrara, Alessandra Pecorelli, Brittany Woodby, Andrew T. Hoyle, Andrew Simonson, and Giuseppe Valacchi

Prolonged and intensive exercise induces transient inflammation, and this has traditionally been measured using a variety of biomarkers, including acute phase proteins, the neutrophil/lymphocyte ratio, and cytokine panels ( Nieman & Wentz, 2019 ; Nieman et al. 2019 ). Emerging evidence indicates

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Nicolette C. Bishop, Michael Gleeson, Ceri W. Nicholas, and Ajmol Ali

Ingesting carbohydrate (CHO) beverages during prolonged, continuous heavy exercise results in smaller changes in the plasma concentrations of several cytokines and attenuates a decline in neutrophil function. In contrast, ingesting CHO during prolonged intermittent exercise appears to have negligible influence on these responses, probably due to the overall moderate intensity of these intermittent exercise protocols. Therefore, we examined the effect of CHO ingestion on plasma interIeukin-6 (IL-6), tumor necrosis factor-α (TNF-α) and lipopolysaccharide (LPS)-stimuIated neutrophil degranulation responses to high-intensity intermittent running. Six trained male soccer players performed 2 exercise trials, 7 days apart, in a randomized, counterbalanced design. On each occasion, they completed six 15-min periods of intermittent running consisting of maximal sprinting interspersed with less intense periods of running and walking. Subjects consumed either CHO or artificially sweetened placebo(PLA) beverages immediately before and at 15-min intervals during the exercise. At 30 min post-exercise, CHO versus PLA was associated with a higher plasma glucose concentration (p< .01), a lower plasma cortisol and IL-6 concentration (p < .02), and fewer numbers of circulating neutrophils (p < .05). Following the exercise, LPS-stimulated elastase release per neutrophil fell 31 % below baseline values on the PLA trial (p = .06) compared with 11% on the CHO trial (p = .30). Plasma TNF-α concentration increased following the exercise (main effect of time, p < .001) but was not affected by CHO. These data indicate that CHO ingestion attenuates changes in plasma IL-6 concentration, neutrophil trafficking, and LPS-stimulated neutrophil degranulation in response to intermittent exercise that involves bouts of very high intensity exercise.

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Elizabeth L. Abbey and Janet Walberg Rankin


This study compared the effect of a honey-sweetened beverage with those of a commercial sports drink and a placebo on performance and inflammatory response to a 90-min soccer simulation.


Ten experienced male soccer players randomly performed 3 trials (honey [H], sports drink [S], and placebo [P]), consuming the beverage before and during halftime for a total of 1.0 g/kg carbohydrate for H and S. Performance measures included 5 sets (T1–T5) of a high-intensity run and agility and ball-shooting tests followed by a final progressive shuttle-run (PSR) test to exhaustion. Blood samples were drawn pretest, posttest (B2), and 1 hr posttest (B3) for markers of inflammation, oxygen radical absorbance capacity (ORAC), and hormone response.


T2–T5 were significantly slower than T1 (p < .05), and a decrease in PSR time was observed from baseline (–22.9%) for all treatments. No significant effect of the interventions was observed for any performance measures. Plasma IL-1ra levels increased posttest for all treatments (65.5% S, 63.9% P, and 25.8% H), but H was significantly less than S at posttest and P at B3. Other cytokines and ORAC increased at B2 (548% IL-6, 514% IL-10, 15% ORAC) with no difference by treatment.


Acute ingestion of honey and a carbohydrate sports drink before and during a soccer-simulation test did not improve performance, although honey attenuated a rise in IL-1ra. Ingestion of carbohydrate and/or antioxidant-containing beverages at frequencies typical of a regulation match may not be beneficial for trained soccer players.

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Piotr Basta, Łucja Pilaczyńska-Szczȩśniak, Donata Woitas-Ślubowska, and Anna Skarpańska-Stejnborn

This investigation examined the effect of supplementation with Biostimine, extract from aloe arborescens Mill. leaves, on the levels of pro-oxidant–antioxidant equilibrium markers and anti- and proinflammatory cytokines in rowers subjected to exhaustive exercise. This double-blind study included 18 members of the Polish Rowing Team. Subjects were randomly assigned to the supplemented group (n = 9), which received one ampoule of Biostimine once daily for 4 weeks, or to the placebo group (n = 9). Subjects performed a 2,000-meter-maximum test on a rowing ergometer at the beginning and end of the preparatory camp. Blood samples were obtained from the antecubital vein before each exercise test, 1 min after completing the test and after a 24-hr recovery period. Superoxide dismutase and glutathione peroxidase activity as well as the concentration of thiobarbituric acid reactive substances (TBARS) were assessed in erythrocytes. In addition, total antioxidant capacity (TAC) and creatine kinase activity were measured in plasma samples, and cytokine (IL-6, IL-10) concentrations were determined in the serum. Before and after Biostimine supplementation, exercise significantly increased the values of SOD, IL-6, IL-10, and TBARS in both groups. However, postexercise and recovery levels of TBARS were significantly lower in athletes receiving Biostimine than in controls. After supplementation, TAC was the only variable with the level being significantly higher in the supplemented group than in the placebo group. Consequently, we can conclude that Biostimine supplementation reduces the postexercise level of TBARS by increasing the antioxidant activity of plasma but has no effect on inflammatory markers.

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Dru A. Henson, David C. Nieman, E. Edward Pistilli, Brian Schilling, AnnaRita Colacino, Allan C. Utter, Omar R. Fagoaga, Debra M. Vinci, and Sandra L. Nehlsen-Cannarella

The influence of 6% carbohydrate ingestion and age on PHA-induced lymphocyte proliferation and in vitro cytokine production was studied in 48 runners following a competitive marathon. Runners were randomly assigned to carbohydrate (C; n = 23) and placebo (P; n = 25) groups, with blood samples taken before, immediately after, and 1.5 hr post-race. C versus P ingestion resulted in higher plasma glucose, lower plasma corlisol, reduced neutrophilia, and mono-cytosis during recovery, but had no effect on the post-exercise reduction in T-lymphocytes or NK cells, or on race times. No group differences were observed for PHA-induced lymphocyte proliferation or cytokine production. However, for all subjects combined, lymphocyte proliferation and IFN-γ secretion decreased significantly below pre-race values by 1.5 hr of recovery, and these were negatively correlated with plasma cortisol. Young (<50 years; n = 36) and old (≥50 years; n = 12) runners exhibited parallel post-race declines in lymphocyte proliferation and IFN-γ secretion, with the older group exhibiting a 33–59% lower proliferation at each time point. In conclusion, PHA-induced lymphocyte proliferation and cytokine production decreased significantly following a marathon, and this decrease was strongly linked to cortisol and only partially linked to T-cell changes. This decrease occurred in both younger and older runners and was not influenced by carbohydrate.