The impact of dietary protein supplementation on recovery from resistance exercise-induced muscle damage: A systematic review with meta-analysis.

BACKGROUND: It is unknown whether dietary protein consumption can attenuate resistance exercise-induced muscle damage (EIMD). Managing EIMD may accelerate muscle recovery and allow frequent, high-quality exercise to promote muscle adaptations. This systematic review and meta-analysis examined the impact of peri-exercise protein supplementation on resistance EIMD. METHODS: A literature search was conducted on PubMed, SPORTDiscus, and Web of Science up to March 2021 for relevant articles. PEDro criteria were used to assess bias within included studies. A Hedges' g effect size (ES) was calculated for indirect markers of EIMD at h post-exercise. Weighted ESs were included in a random effects model to determine overall ESs over time. RESULTS: Twenty-nine studies were included in the systematic review and 40 trials were included in ≥1 meta-analyses (16 total). There were significant overall effects of protein for preserving isometric maximal voluntary contraction (MVC) at 96 h (0.563 [0.232, 0.894]) and isokinetic MVC at 24 h (0.639 [0.116, 1.162]), 48 h (0.447 [0.104, 0.790]), and 72 h (0.569 [0.136, 1.002]). Overall ESs were large in favour of protein for attenuating creatine kinase concentration at 48 h (0.836 [-0.001, 1.673]) and 72 h (1.335 [0.294, 2.376]). Protein supplementation had no effect on muscle soreness compared with the control. CONCLUSION: Peri-exercise protein consumption could help maintain maximal strength and lower creatine kinase concentration following resistance exercise but not reduce muscle soreness. Conflicting data may be due to methodological divergencies between studies. Standardised methods and data reporting for EIMD research are needed.

The effects of collagen peptide supplementation on body composition, collagen synthesis, and recovery from joint injury and exercise: a systematic review.

Collagen peptide supplementation (COL), in conjunction with exercise, may be beneficial for the management of degenerative bone and joint disorders. This is likely due to stimulatory effects of COL and exercise on the extracellular matrix of connective tissues, improving structure and load-bearing capabilities. This systematic review aims to evaluate the current literature available on the combined impact of COL and exercise. Following Preferred Reporting Items for Systematic Reviews and Meta-analyses guidelines, a literature search of three electronic databases-PubMed, Web of Science and CINAHL-was conducted in June 2020. Fifteen randomised controlled trials were selected after screening 856 articles. The study populations included 12 studies in recreational athletes, 2 studies in elderly participants and 1 in untrained pre-menopausal women. Study outcomes were categorised into four topics: (i) joint pain and recovery from joint injuries, (ii) body composition, (iii) muscle soreness and recovery from exercise, and (iv) muscle protein synthesis (MPS) and collagen synthesis. The results indicated that COL is most beneficial in improving joint functionality and reducing joint pain. Certain improvements in body composition, strength and muscle recovery were present. Collagen synthesis rates were elevated with 15 g/day COL but did not have a significant impact on MPS when compared to isonitrogenous higher quality protein sources. Exact mechanisms for these adaptations are unclear, with future research using larger sample sizes, elite athletes, female participants and more precise outcome measures such as muscle biopsies and magnetic imagery.

Peripheral EphrinB1/EphB1 signalling attenuates muscle hyperalgesia in MPS patients and a rat model of taut band-associated persistent muscle pain.

BACKGROUND: Myofascial pain syndrome (MPS) is an important clinical condition that is characterized by chronic muscle pain and a myofascial trigger point (MTrP) located in a taut band (TB). Previous studies showed that EphrinB1 was involved in the regulation of pathological pain via EphB1 signalling, but whether EphrinB1-EphB1 plays a role in MTrP is not clear. METHODS: The present study analysed the levels of p-EphB1/p-EphB2/p-EphB3 in biopsies of MTrPs in the trapezius muscle of 11 MPS patients and seven healthy controls using a protein microarray kit. EphrinB1-Fc was injected intramuscularly to detect EphrinB1s/EphB1s signalling in peripheral sensitization. We applied a blunt strike to the left gastrocnemius muscles (GM) and eccentric exercise for 8 weeks with 4 weeks of recovery to analyse the function of EphrinB1/EphB1 in the muscle pain model. RESULTS: P-EphB1, p-EphB2, and p-EphB3 expression was highly increased in human muscles with MTrPs compared to healthy muscle. EphB1 (r = 0.723, n = 11, P < 0.05), EphB2 (r = 0.610, n = 11, P < 0.05), and EphB3 levels (r = 0.670, n = 11, P < 0.05) in the MPS group were significantly correlated with the numerical rating scale (NRS) in the MTrPs. Intramuscular injection of EphrinB1-Fc produces hyperalgesia, which can be partially prevented by pre-treatment with EphB1-Fc. The p-EphB1 contents in MTrPs of MPS animals were significantly higher than that among control animals (P < 0.01). Intramuscular administration of the EphB1 inhibitor EphB1-Fr significantly suppressed mechanical hyperalgesia. CONCLUSIONS: The present study showed that the increased expression of p-EphB1/p-EphB2/p-EphB3 was related to MTrPs in patients with MPS. This report is the first study to examine the function of EphrinB1-EphB1 signalling in primary muscle afferent neurons in MPS patients and a rat animal model. This pathway may be one of the most important and promising targets for MPS.

One week of magnesium supplementation lowers IL-6, muscle soreness and increases post-exercise blood glucose in response to downhill running.

PURPOSE: Magnesium supplementation modulates glucose metabolism and inflammation, which could influence exercise performance and recovery. This study investigated the effect of magnesium intake on physiological responses and performance during eccentric exercise and recovery. METHODS: Nine male recreational runners completed a counterbalanced, double-blind, placebo-controlled, cross-over study, registered at ClinicalTrial.gov. Participants consumed low magnesium diets and were supplemented with 500 mg/day of magnesium (SUP) or placebo (CON) for 7 days prior to a 10 km downhill (- 10%) running time trial (TT), separated by a 2-week washout period. At baseline and 24 h post-TT, maximal muscle force was measured. Interleukin-6 (IL-6), soluble interleukin-6 receptor (sIL-6R) and creatine kinase (CK) were measured at rest, 0 h, 1 h and 24 h post-TT. Muscle soreness was measured at the previous times plus 48 h and 72 h post. Glucose and lactate were measured during the TT. RESULTS: The main effect of condition was detected for IL-6 (SUP: 1.36 ± 0.66 vs CON: 2.06 ± 1.14 pg/ml) (P < 0.05, η2 = 0.54), sIL-6R (SUP: 27,615 ± 8446 vs CON: 24,368 ± 7806 pg/ml) (P < 0.05, η2 = 0.41) and muscle soreness (P < 0.01, η2 = 0.67). Recovery of blood glucose and muscle soreness were enhanced in SUP post-TT. There were no differences in glucose and lactate during the TT, or post measures of CK and maximal muscle force. CONCLUSION: Magnesium supplementation reduced the IL-6 response, enhanced recovery of blood glucose, and muscle soreness after strenuous exercise, but did not improve performance or functional measures of recovery.

Fibromyalgia and nutrition: Therapeutic possibilities?

Fibromyalgia (FM) is a complex chronic condition of unknown etiology, characterized by deep and widespread pain, sleep problems, cognitive impairment, fatigue, and other well-known functional symptoms. Recently, it has been proposed that an imbalance of nutritive components, including essential metal ions and vitamins, might play a critical role in the development of FM. Muscle pain has been associated with deficiencies in amino acids, magnesium, selenium, vitamins B and D, as well as with the harmful effects of heavy metals, such as mercury, cadmium, and lead. Research indicates that patients deficient in certain essential nutrients may develop dysfunction of pain inhibitory mechanisms together with fatigue and other FM symptoms. Additionally, mercury and other toxic elements may interfere with the bioavailability of essential nutrients. This review examines the many effects of metals and vitamins in pain evaluation of FM patients. Dietary guidance is therefore critical for FM patients to help them in correcting a suboptimal or deficient intake of essential nutrients. When optimal levels of nutrition are achieved, pain levels are usually lowered. Additional research is recommended in the field of FM and nutrition to disclose further possible relationships.

Arachidonic acid supplementation transiently augments the acute inflammatory response to resistance exercise in trained men.

Strenuous exercise can result in skeletal muscle damage, leading to the systemic mobilization, activation, and intramuscular accumulation of blood leukocytes. Eicosanoid metabolites of arachidonic acid (ARA) are potent inflammatory mediators, but whether changes in dietary ARA intake influence exercise-induced inflammation is not known. This study investigated the effect of 4 wk of dietary supplementation with 1.5 g/day ARA ( n = 9, 24 ± 1.5 yr) or corn-soy oil placebo ( n = 10, 26 ± 1.3 yr) on systemic and intramuscular inflammatory responses to an acute bout of resistance exercise (8 sets each of leg press and extension at 80% one-repetition maximum) in previously trained men. Whole EDTA blood, serum, peripheral blood mononuclear cells (PMBCs), and skeletal muscle biopsies were collected before exercise, immediately postexercise, and at 2, 4, and 48 h of recovery. ARA supplementation resulted in higher exercise-stimulated serum creatine kinase activity [incremental area under the curve (iAUC) P = 0.046] and blood leukocyte counts (iAUC for total white cells, P < 0.001; neutrophils: P = 0.007; monocytes: P = 0.015). The exercise-induced fold change in peripheral blood mononuclear cell mRNA expression of interleukin-1ß ( IL1B), CD11b ( ITGAM), and neutrophil elastase ( ELANE), as well as muscle mRNA expression of the chemokines interleukin-8 ( CXCL8) and monocyte chemoattractant protein 1 ( CCL2) was also greater in the ARA group than placebo. Despite this, ARA supplementation did not influence the histological presence of leukocytes within muscle, perceived muscle soreness, or the extent and duration of muscle force loss. These data show that ARA supplementation transiently increased the inflammatory response to acute resistance exercise but did not impair recovery. NEW & NOTEWORTHY Daily arachidonic acid supplementation for 4 wk in trained men augmented the acute systemic and intramuscular inflammatory response to a subsequent bout of resistance exercise. Greater exercise-induced inflammatory responses in men receiving arachidonic acid supplementation were not accompanied by increased symptoms of exercise-induced muscle damage. Although increased dietary arachidonic acid intake does not appear to influence basal inflammation in humans, the acute inflammatory response to exercise stress is transiently increased following arachidonic acid supplementation.

Effect of tart cherry juice on recovery and next day performance in well-trained Water Polo players.

BACKGROUND: Tart Montmorency cherries contain high concentrations of phytochemicals and anthocyanins, which have recently been linked to improved athletic recovery and subsequent performance. To date however, previous work reporting promising results has focused on land-based endurance sports, with any potential benefits to team sports remaining unknown. As such, this investigation set-out to examine the effect of supplemental tart cherry juice (CJ) on recovery and next day athletic performance in highly-trained water-based team sport athletes over seven days. METHODS: In a randomised, double-blind, repeated measures, crossover design, nine male Water Polo athletes were supplemented with CJ or a placebo equivalent (PLA) for six consecutive days. Prior to, and at the completion of the supplementation period, water-based performance testing was conducted. On day 6, participants also undertook a fatiguing simulated team game activity. Venous blood samples were collected (Pre-exercise: day 1, 6 and 7; Post-exercise: day 6) to investigate markers of inflammation [Interleukin-6 (IL-6); C-reactive protein (CRP)] and oxidative stress [Uric Acid (UA); F2-Isoprostane (F2-IsoP)]. A daily diary was also completed (total quality of recovery, delayed onset muscle soreness) as a measure of perceptual recovery. RESULTS: In both conditions, day 6 post-exercise IL-6 was significantly higher than pre-exercise and day 7 (p < 0.05); CRP was greater on day 7 as compared to day 6 pre- and post-exercise (p < 0.05); F2-IsoP was significantly lower on day 7 as compared to day 1 and day 6 (p < 0.05); UA remained unchanged (p > 0.05). No differences were found for any performance or recovery measures. CONCLUSIONS: The lack of difference observed in the blood markers between groups may reflect the intermittent, non-weight bearing demands of Water Polo, with such activity possibly unable to create a substantial inflammatory response or oxidative stress (over 7 days) to impede performance; thereby negating any potential beneficial effects associated with CJ supplementation. TRIAL REGISTRATION: This trial was registered with the Australian and New Zealand Clinical Trials Registry (ANZCTR). Registration number: ACTRN12616001080415. Date registered: 11/08/2016, retrospectively registered.

Post-exercise branched chain amino acid supplementation does not affect recovery markers following three consecutive high intensity resistance training bouts compared to carbohydrate supplementation.

BACKGROUND: Amino acid supplementation has been shown to potentially reduced exercise-induced muscle soreness. Thus, the purpose of this study was to examine if branched chain amino acid and carbohydrate (BCAACHO) versus carbohydrate-only sports drink (CHO) supplementation attenuated markers of muscle damage while preserving performance markers following 3 days of intense weight training. METHODS: Healthy resistance-trained males (n = 30) performed preliminary testing (T1) whereby they: 1) donated a baseline blood draw, 2) performed knee extensor dynamometry to obtain peak quadriceps isometric and isokinetic torque as well as electromyography (EMG) activity at 60°/s and 120°/s, and 3) performed a one repetition maximum (1RM) barbell back squat. The following week participants performed 10 sets x 5 repetitions at 80 % of their 1RM barbell back squat for 3 consecutive days and 48 h following the third lifting bout participants returned for (T2) testing whereby they repeated the T1 battery. Immediately following and 24 h after the three lifting bouts, participants were randomly assigned to consume one of two commercial products in 600 mL of tap water: 1) BCAAs and CHO (3 g/d L-leucine, 1 g/d L-isoleucine and 2 g/d L-valine with 2 g of CHO; n = 15), or 2) 42 g of CHO only (n = 15). Additionally, venous blood was drawn 24 h following the first and second lifting bouts and 48 h following the third bout to assess serum myoglobin concentrations, and a visual analog scale was utilized prior, during, and after the 3-d protocol to measure subjective perceptions of muscular soreness. RESULTS: There were similar decrements in 1RM squat strength and isokinetic peak torque measures in the BCAA-CHO and CHO groups. Serum myoglobin concentrations (p = 0.027) and perceived muscle soreness (p < 0.001) increased over the intervention regardless of supplementation. A group*time interaction was observed for monocyte percentages (p = 0.01) whereby BCAA-CHO supplementation attenuated increases in this variable over the duration of the protocol compared to CHO supplementation. CONCLUSION: BCAA-CHO supplementation did not reduce decrements in lower body strength or improve select markers of muscle damage/soreness compared to CHO supplementation over three consecutive days of intense lower-body training.

Effects of combined ß-hydroxy-ß-methylbutyrate (HMB) and whey protein ingestion on symptoms of eccentric exercise-induced muscle damage.

BACKGROUND: The purpose of this study was to examine the effects of combined ß-hydroxy-ß-methylbutyrate (HMB) and whey protein ingestion on muscle strength and damage following a single bout of eccentric exercise. METHODS: Eighteen untrained male subjects were assigned to HMB and Whey protein (HMB + Whey; 3 g/day HMB and 36.6 g/day whey protein, n = 6), HMB (3 g/day, n = 6), or whey protein (36.6 g/day, n = 6) groups. Ingestion commenced 7 days before non-dominant elbow flexor eccentric exercise (30 deg/sec, 6 reps × 7 sets) and continued until 4 days post-exercise. The maximal isometric strength, muscle soreness, plasma creatine kinase (CK), lactate dehydrogenase (LDH) were assessed pre-exercise, and at 1, 2, 3, and 5 days after exercise. RESULTS: The change scores of maximal isometric strength significantly decreased at day 1, 2, and 5 in the whey protein group compared to pre value and that in HMB + Whey protein and HMB groups decreased at day 1 and 5. The muscle soreness significantly increased in the whey and HMB + Whey protein groups at day 3 compared to pre value (p < 0.05). CK and LDH significantly increased (time effect: p < 0.05) after exercise. However, all data were not significant difference among the groups. CONCLUSIONS: These results suggest that ingestion of combined HMB and whey protein does not have a role to inhibit muscle strength loss and soreness, and decrease in muscle damage markers after eccentric exercise in comparison with HMB and whey protein alone.

Efficacy of massage on muscle soreness, perceived recovery, physiological restoration and physical performance in male bodybuilders.

It is believed that sport massage after intensive exercise might improve power and perceptual recovery in athletes. However, few studies have been done in this area. This study aimed to examine the effect of massage on the performance of bodybuilders. Thirty experienced male bodybuilders were randomly assigned to either a massage group (n = 15) or a control group (n = 15). Both groups performed five repetition sets at 75-77% of 1RM of knee extensor and flexor muscle groups. The massage group then received a 30-min massage after the exercise protocol while the control group maintained their normal passive recovery. Criteria under investigation included: plasma creatine kinase (CK) level, agility test, vertical jump test, isometric torque test, and perception of soreness. All variables were measured over 6 time periods: baseline, immediately after the DOMS inducing protocol, right after the massage, and 24, 48, and 72 h after the massage. Both groups showed significant (P < .001) decreases in jumping, agility performance, and isometric torque, but significant (P < .001) increases in CK and muscle soreness levels. The massage group in general demonstrated a better recovery rate. As such, a post-exercise massage session can improve the exercise performance and recovery rate in male bodybuilders after intensive exercise.

No differential effects of divergent isocaloric supplements on signaling for muscle protein turnover during recovery from muscle-damaging eccentric exercise.

Unaccustomed high-intensity eccentric exercise (ECC) can provoke muscle damage including several days of muscle force loss. Post-exercise dietary supplementation may provide a strategy to accelerate rate of force regain by affecting mechanisms related to muscle protein turnover. The aim of the current study was to investigate if protein signaling mechanisms involved in muscle protein turnover would be differentially affected by supplementation with either whey protein hydrolysate and carbohydrate (WPH+CHO) versus isocaloric carbohydrate (CHO) after muscle-damaging ECC. Twenty-four young healthy participants received either WPH+CHO (n = 12) or CHO supplements (n = 12) during post-exercise recovery from 150 maximal unilateral eccentric contractions. Prior to, at 3 h and at 24, 48, 96 and/or 168 h post-exercise, muscle strength, muscle soreness, and Akt-mTOR and FOXO signaling proteins, were measured in an ECC exercising leg and in the contralateral non-exercise control leg (CON). After ECC, muscle force decreased by 23-27 % at 24 h post-exercise, which was followed by gradual, although not full recovery at 168 h post-exercise, with no differences between supplement groups. Phosphorylation of mTOR, p70S6K and rpS6 increased and phosphorylation of FOXO1 and FOXO3 decreased in the ECC leg, with no differences between supplement groups. Phosphorylation changes were also observed for rpS6, FOXO1 and FOXO3a in the CON leg, suggesting occurrence of remote tissue effects. In conclusion, divergent dietary supplementation types did not produce differences in signaling for muscle turnover during recovery from muscle-damaging exercise.

Impaired mitochondrial function and reduced energy cost as a result of muscle damage.

PURPOSE: Although it has been largely acknowledged that isometric neuromuscular electrostimulation (NMES) exercise induces larger muscle damage than voluntary contractions, the corresponding effects on muscle energetics remain to be determined. Voluntary exercise-induced muscle damage (EIMD) has been reported to have minor slight effects on muscle metabolic response to subsequent dynamic exercise, but the magnitude of muscle energetics alterations for NMES EIMD has never been documented. METHODS: ³¹P magnetic resonance spectroscopy measurements were performed in 13 young healthy males during a standardized rest-exercise-recovery protocol before (D0) and 2 d (D2) and 4 d (D4) after NMES EIMD on knee extensor muscles. Changes in kinetics of phosphorylated metabolite concentrations (i.e., phosphocreatine [PCr], inorganic phosphate [Pi], and adenosine triphosphate [ATP]) and pH were assessed to investigate aerobic and anaerobic rates of ATP production and energy cost of contraction (Ec). RESULTS: Resting [Pi]/[PCr] ratio increased at D2 (+39%) and D4 (+29%), mainly owing to the increased [Pi] (+43% and +32%, respectively), whereas a significant decrease in resting pH was determined (-0.04 pH unit and -0.03 pH unit, respectively). PCr recovery rate decreased at D2 (-21%) and D4 (-23%) in conjunction with a significantly decreased total rate of ATP production at D4 (-18%) mainly owing to an altered aerobic ATP production (-19%). Paradoxically, Ec was decreased at D4 (-21%). CONCLUSION: Overall, NMES EIMD led to intramuscular acidosis in resting muscle and mitochondrial impairment in exercising muscle. Alterations of noncontractile processes and/or adaptive mechanisms to muscle damage might account for the decreased Ec during the dynamic exercise.

Preventive effects of 10-day supplementation with saffron and indomethacin on the delayed-onset muscle soreness.

OBJECTIVE: Delayed-onset muscle soreness (DOMS) often occurs after unaccustomed eccentric exercise and reduces exercise performance. We aimed to study the preventive effects of saffron and indomethacin on the biochemical and functional indicators of DOMS after 1-session eccentric exercise. DESIGN: A 10-day, randomized, double-blind, placebo-controlled, pretest-posttest design. SETTING: Controlled research laboratory. PARTICIPANTS: Thirty-nine nonactive male university students randomly divided into saffron (n = 12), indomethacin (n = 12), and control (n = 15) groups. INTERVENTIONS: Saffron group received 1 capsule containing dried saffron powder (n = 12, 300 mg/d), indomethacin group received 75 mg indomethacin (n = 12, 25 mg thrice a day), and control group (n = 15) received placebo capsules, 1 week before and 3 days after eccentric exercise. Ten days before and 24, 48, and 72 hours after muscle soreness protocol, the maximum isometric and isotonic forces, plasma creatine kinase (CK), plasma lactate dehydrogenase (LDH), perceived pain, knee range of movement, and thigh circumference were measured. Muscle soreness protocol was performed with a weight load equal to 80% of the maximum isotonic force in 4 sessions with 20 repetitions and 3-minute rest in between. MAIN OUTCOME MEASURES: This study shows that 10-day supplementation with 300 mg saffron significantly decreased the CK and LDH concentrations (P < 0.0001). In the saffron group, there was no decline in maximum isometric and isotonic forces after eccentric exercise, but a significant decline in the isometric force was observed in the control group (P < 0.0001). No pain was reported in the saffron group, whereas the indomethacin group experienced pain before 72 hours (P < 0.001). CONCLUSIONS: Results obtained from the current novel research indicate a strong preventive effect of 10-day supplementation with saffron on the DOMS. CLINICAL RELEVANCE: The saffron can be used to prevent DOMS and alleviate the DOMS symptoms.

A 7-day oral supplementation with branched-chain amino acids was ineffective to prevent muscle damage during a marathon.

The aim of this study was to determine the effectiveness of a 7-day oral supplementation with branched-chain amino acids (BCAA) to prevent muscle damage during a marathon. Forty-six experienced runners were randomly divided into two groups, one with BCAA supplementation (n = 25, supplemented with 5 g day(-1) of powdered 1:0.5:0.5 leucine:isoleucine:valine, during the 7 days prior to the competition) and the other as a control group (n = 21, supplemented with an isocaloric placebo). Before the marathon race and within 3 min of finishing, leg muscle power was measured with a maximal countermovement jump and a urine sample was obtained. During the race, running pace was measured by means of a time-chip. Myoglobin concentration was determined in the urine samples as an indirect marker of muscle damage. A visual analog scale (0-10 points) was used to assess leg muscle pain during the race. In the BCAA group, the mean running pace during the marathon was similar to the control group (3.3 ± 0.4 vs. 3.3 ± 0.5 m s(-1), respectively, 0.98). The pre- to post-race reduction in muscle power was similar in both BCAA and control groups (-23.0 ± 16.1 vs. -17.3 ± 13.8 %, P = 0.13). Post-race urine myoglobin concentration was similar in both BCAA and control groups (5.4 ± 7.5 vs. 4.5 ± 8.6 µg mL(-1), P = 0.70). Finally, there were no differences between groups in the perceived muscle pain during the race (6 ± 1 vs. 5 ± 1 points, P = 0.80). A 7-day supplementation of BCAA (5 g day(-1)) did not increase the running performance during a marathon. Furthermore, BCAA supplementation was ineffective to prevent muscle power loss, muscle damage or perceived muscle pain during a marathon race.

Comparative metabolomics of muscle interstitium fluid in human trapezius myalgia: an in vivo microdialysis study.

PURPOSE: The mechanisms behind trapezius myalgia are unclear. Many hypotheses have been presented suggesting an altered metabolism in the muscle. Here, muscle microdialysate from healthy and myalgic muscle is analysed using metabolomics. Metabolomics analyse a vast number of metabolites, enabling a comprehensive explorative screening of the cellular processes in the muscle. METHODS: Microdialysate samples were obtained from the shoulder muscle of healthy and myalgic subjects that performed a work and stress test. Samples from the baseline period and from the recovery period were analysed using gas chromatography­mass spectrometry (GC­MS) together with multivariate analysis to detect differences in extracellular content of metabolites between groups. Systematic differences in metabolites between groups were identified using multivariate analysis and orthogonal partial least square discriminate analysis (OPLS-DA). A complementary Mann­Whitney U test of group difference in individual metabolites was also performed. RESULTS: A large number of metabolites were detected and identified in this screening study. At baseline, no systematic differences between groups were observed according to the OPLS-DA. However, two metabolites, l-leucine and pyroglutamic acid, were significantly more abundant in the myalgic muscle compared to the healthy muscle. In the recovery period, systematic difference in metabolites between the groups was observed according to the OPLS-DA. The groups differed in amino acids, fatty acids and carbohydrates. Myristic acid and putrescine were significantly more abundant and beta-d-glucopyranose was significantly less abundant in the myalgic muscle. CONCLUSION: This study provides important information regarding the metabolite content, thereby presenting new clues regarding the pathophysiology of the myalgic muscle.