Nutrients. 2016 Jul; 8(7): 441. 
Published online 2016 Jul 22.   doi:  10.3390/nu8070441
PMCID: PMC4963917
Abstract
This
 study investigated Montmorency tart cherry concentrate (MC) 
supplementation on markers of recovery following prolonged, intermittent
 sprint activity. Sixteen semi-professional, male soccer players, who 
had dietary restrictions imposed for the duration of the study, were 
divided into two equal groups and consumed either MC or placebo (PLA) 
supplementation for eight consecutive days (30 mL twice per day). On day
 5, participants completed an adapted version of the Loughborough 
Intermittent Shuttle Test (LISTADAPT). Maximal voluntary 
isometric contraction (MVIC), 20 m Sprint, counter movement jump (CMJ), 
agility and muscle soreness (DOMS) were assessed at baseline, and 24, 48
 and 72 h post-exercise. Measures of inflammation (IL-1-β, IL-6, IL-8, 
TNF-α, hsCRP), muscle damage (CK) and oxidative stress (LOOH) were 
analysed at baseline and 1, 3, 5, 24, 48 and 72 h post-exercise. 
Performance indices (MVIC, CMJ and agility) recovered faster and muscle 
soreness (DOMS) ratings were lower in the MC group (p < 
0.05). Additionally, the acute inflammatory response (IL-6) was 
attenuated in the MC group. There were no effects for LOOH and CK. These
 findings suggest MC is efficacious in accelerating recovery following 
prolonged, repeat sprint activity, such as soccer and rugby, and lends 
further evidence that polyphenol-rich foods like MC are effective in 
accelerating recovery following various types of strenuous exercise.
Keywords: recovery, strenuous exercise, muscle damage, prunus cerasus, functional foods
1. Introduction
Prolonged, field-based, intermittent sprint sports are popular across the world at both elite and recreational levels [1].
 Sports, such as soccer, field hockey and rugby, require a high volume 
of energy turnover and eccentric muscle actions resulting in metabolic 
and mechanically induced stress. Indeed, soccer play results in elevated
 post-match inflammatory [2,3], oxidative stress [2,4,5] and muscle damage [2,4] markers, as well as decrements in physical performance [2,4,5].
 The Loughborough Intermittent Shuttle Test (LIST), which closely 
simulates activity patterns, and the physiological and metabolic demands
 of soccer [6], has been shown to incur similar stress responses [7,8,9],
 and, thus, provides a tool to induce similar physiological stress to 
game-play, but in a controlled environment. In light that athlete 
schedules require training or competition on multiple occasions within a
 few days, the importance of recovery strategies when preparing for the 
next game or training session is critical [10]. This is especially pertinent given that recovery is incomplete at 48 h following actual [2,11] and simulated [9] match-play.
Montmorency tart cherries have been shown on numerous occasions to be of benefit in exercise recovery [12,13,14,15,16],
 which have been proposed to be as a result of the high concentrations 
of phytochemicals, and in particular, the flavanoids anthocyanins [14,17,18,19].
 These compounds can reduce oxidative stress and been shown to be a 
cyclooxygenase inhibitor (COX), to a similar extent as NSAIDs [20,21]. A series of studies have investigated the use of Montmorency cherries in influencing recovery from running [14,22], heavy eccentric contractions [15,16] and cycling [12,13].
 Collectively, these lines of investigation suggest that Montmorency 
cherries could also be applied to aiding recovery following high 
intensity concurrent sports that incorporate and very high metabolic 
component that is accompanied by high intensity eccentric contractions. 
Given
 that Montmorency cherries have been shown to be of benefit in exercise 
recovery following high intensity eccentric contractions and 
metabolically challenging exercise, it makes the expectation tenable 
that it could be applied to sports of a concurrent nature. Therefore the
 aim of this study was to investigate the effect of Montmorency cherries
 on recovery indices following a protocol designed to replicate the 
physiological demands of prolonged intermittent sprint activity such as 
those seen in field based sports. It was hypothesised that MC 
supplementation would attenuate post-exercise inflammatory and oxidative
 stress responses, and aid the return of functional performance.
2. Methods
2.1. Participants
Sixteen
 semi-professional (step 5 and above in the Football Association 
National pyramid, UK), male soccer players (mean ± SD age, height, mass,
 predicted V̇O2max was 25 ± 4 years; 180.8 ± 7.4 cm, 81.9 ± 6.6 kg, 54.9 mL·kg−1·min−1,
 respectively) volunteered to take part in the study. All procedures 
were granted Ethical clearance by the University’s Research Ethics 
Committee prior to testing and were conducted in accordance with the 
Helsinki Declaration. Inclusion criteria required participants to have 
trained in soccer consistently across the preceding 3 years and be free 
of any lower limb injury for the preceding 6 months. This was assessed 
through the completion of training history and health screening 
questionnaires. Following both verbal and written briefings on the 
requirements of the study, written informed consent was collected from 
all participants.
2.2. Study Design
A
 double blind, placebo controlled design with independent groups design 
was employed. Participants attended the laboratory on six separate 
occasions across a period of no longer than 15 days (Figure 1). On visit 1, participants completed a multi-stage shuttle test [23] in order to predict VO2max,
 which was followed by familiarisation with a battery of functional 
performance tests and one 15 min section of the Loughborough 
Intermittent Shuttle Test Part A (LIST) [6].
 Participants were then randomly but equally assigned to either a 
Montmorency cherry (MC) or placebo (PLA) group, matched by predicted VO2max score (54.3 vs. 55.4 mL·kg−1·min−1).
Participants
 returned to the laboratory for visit 2 within 5 days to complete the 
battery of baseline functional measures that followed a standardised 
warm up; these were countermovement jump height (CMJ), 20 m sprint time 
(20 m), MVIC of the knee extensors, agility (5-0-5), which were preceded
 by assessment of active muscle soreness (DOMS). Participants were then 
provided with 24 MC or PLA beverages in a double blind manner along with
 verbal and written instructions on how to consume the beverages. They 
were also provided with a diet record diary and a list of foods to avoid
 throughout the 4 days prior to and during the trial period. During the 4
 day period leading up to the trial day, participants were contacted and
 instructed to begin supplementation and dietary restrictions.
Visits
 3–6 commenced at 8:00 a.m. in order to account for diurnal variation. 
On visit 3, participants were required to complete an adapted version of
 the LIST (LISTADAPT). Following the standardised warm-up, 
participants completed a series of 12 × 20 m sprints with a 10 m 
‘stopping zone’, departing every 60 s. This addition was included 
because (1) the LIST protocol does not account for the many bounding, 
leaping and directional changes that are associated with team sports 
play; and (2) previous work from our laboratory has shown only moderate 
responses with regards to the magnitude of stress response following the
 LIST [9].
 A secondary adaptation to the LIST protocol was the completion of 6 × 
15 min sections from the LIST Part A, as opposed to 5 × 15 min sections 
detailed in the original protocol [6].
 This section was included to standardise the distance covered by the 
two groups (in the original protocol, LIST Part B required a run to 
exhaustion, potentially resulting in group differences). During the LISTADAPT participants were provided with water ad libitum.
Visits
 4–6 took place at 24, 48 and 72 h following the start of visit 3 and 
required participants to complete the functional performance test 
battery outlined in visit 2. Venous blood samples were collected at 
baseline (prior to muscle soreness and performance test), immediately 
pre-trial, immediately post-trial and 1, 3, 5, 24, 48 and 72 h 
post-trial for markers of inflammation, oxidative stress and muscle 
damage.
2.3. Supplementation
The
 MC or PLA supplementation was provided to participants after the 
initial visit along with instructions detailing the dosing schedule (30 
mL twice per day, (8:00 a.m., 6:00 p.m.), 7 consecutive days (4 days 
pre- and on each trial day [13])).
 Supplements were prepared by mixing each dose with 100 mL of water 
prior to consumption. The MC was a commercially available Montmorency 
cherry concentrate (CherryActive, Sunbury, UK); previous work from our 
laboratory has shown that the MC used in this study contains a total 
anthocyanin content of 73.5 mg·L−1 of cyanidin-3-glucoside, a total phenolic content of 178.8 gallic acid equivalent·L−1 and an antioxidant capacity (TEAC) of 0.58 trolox equivalents·L−1 [24].
 A commercially available, less than 5% fruit, cordial, mixed with water
 and maltodextrin (MyProtein Ltd, Northwich, UK) until matched for 
energy content of the MC (102 kcal) was used for the PLA supplement. All
 supplements were prepared by an independent member of the department 
prepared in opaque bottles in order to maintain the double blind design.
2.4. Dietary and Exercise Restrictions
Participants
 were instructed to follow a low polyphenolic diet in the 48 h prior to 
the beginning of each MC or PLA supplementation and throughout the 
experimental phase of each study. A list of foods to avoid was provided 
and compliance was assessed through the completion of daily food diaries
 which has been successfully implemented in previous research [25].
 This control measure was used to provide a washout period of 
polyphenols to enable the efficacy of the phenolic-rich cherry 
concentrate intervention. In addition, participants were instructed to 
abstain from any exercise that was not a part of the protocol, 
throughout the same time periods. Lastly, participants attended all 
exercise trials following an overnight fast. These measures ensured that
 dependant variable changes from baseline were likely to be in response 
to the supplementation and the exercise trials implemented within each 
study.
2.5. Functional Performance Tests
The
 functional performance test battery was performed in the following 
order on each occasion: Active muscle soreness assessment (DOMS), 20 m 
sprint, 5-0-5 agility (CV 2.8% [26]),
 countermovement jump (CV 1.9%), knee extensors (repeatability in 
Chapter 3.4). Timings were kept consistent throughout all functional 
performance tests, each test was performed 3 times (excluding DOMS), 
with a 1 min rest between repetitions and 3 min rest between tests.
Delayed
 onset of muscle soreness (DOMS) in the lower limbs was assessed using a
 200 mm visual analogue scale (VAS) with ‘no soreness’ at one end and 
‘unbearably painful’ at the other. On each occasion the VAS was used, 
participants were instructed to place their hands on hips, squat down to
 ~90°, before standing up and immediately making a mark on the scale 
consistent with their perceived soreness. 
Sprint performance (20 m; coefficient of variation (CV) 0.9%) and the 5-0-5 agility test [27,28]
 (CV 2.8%) were assessed using wireless telemetry and infra-red timing 
gates (Brower Timing Systems, Draper, UT, USA) on an indoor athletics 
track. Countermovement jump height (CV 1.9%) was assessed using a jump 
mat (Just Jump, Probotics Inc., Huntsville, AL, USA); participants were 
instructed to stand on the jump mat with their feet parallel and 
approximately shoulder width apart. Following this, participants 
completed a maximal vertical jump whilst maintaining hands on hips 
through flight and landing. MVIC of the dominant knee extensors was 
determined using a strain gauge (MIE Medical Research Ltd., Leeds, UK) 
using the methods described previously [13]. The peak performance from each trial was used for data analysis.
2.6. Blood Sampling
Blood
 samples (35 mL) were collected from a forearm vein located in the 
antecubital fossa region in order to assess for markers of muscle damage
 (creatine kinase [CK]), inflammation (interleukin-1-beta (IL-1-β), 
interleukin-6 (IL-6), tumour necrosis factor-alpha (TNF-α), 
high-sensitivity C-reactive protein (hsCRP)) and oxidative stress (lipid
 hydroperoxides (LOOH)) using previously described methods [12,13];
 intra-sample CVs ranged from 0.7% to 6.8%. Samples were collected into 
serum gel, ethylenediaminetetraacetic acid (EDTA) or sodium heparin 
treated tubes (Vacutainer®BD UK Ltd., Oxford, UK). Samples 
were then immediately centrifuged (Allegra X-22 Centrifuge, Beckman 
Coulter, Bucks, UK) at 2400× g at 4 °C for 15 min before having
 the supernatant removed and stored in aliquots. Aliquots were then 
immediately stored at −80 °C and subsequently analysed for the 
respective indices in each study. 
2.7. Statistical Analysis
All
 data analyses were conducted using IBM SPSS Statistics 20 for Windows 
(Surrey, UK) and are reported as mean ± standard deviation. All data was
 confirmed as parametric via a Shapiro-Wilk test for normality. 
Differences between blood marker variables were analysed by using a 
group (MC vs. PLA) by time-point (Pre-supplement, Post-supplement, 1, 3,
 5, 24, 48 and 72 h) mixed model ANOVA. Functional performance measures 
were analysed using the same model, however with four fewer levels 
(Pre-supplement, Post-supplement, 24, 48 and 72 h). Where significant 
group baseline differences were apparent (MVIC, CMJ, 5-0-5 agility, 20 m
 sprint, DOMS) results were normalised to baseline values prior to 
subsequent statistical analysis. Mauchley’s Test of Sphericity was used 
to assess homogeneity of data and where violations were present, 
Greenhouse-Geiser adjustments were made. Where necessary, interaction 
effects were assessed using LSD post hoc analysis. Prior to all 
analyses, a significance level of p < 0.05 was set.
3. Results
MVIC (Figure 2) showed significant time (F(1,4) = 6.586, p = 0.001, η2  = 0.320), group (F(1,2) = 19.445, p = 0.001, η2  = 0.582) and interaction (F(1,4) = 8.970, p < 0.001, η2 
 = 0.391) effects when data was normalised to baseline values. The 
decline in MIVC performance was not evident in the MC group whereas as 
function had not returned to basal levels at 72 h in the PLA. The peak 
difference occurred at 48 h where MVIC scores in the MC group were found
 to be 19% higher.

Changes in maximum voluntary isometric contraction (MVIC; Panel (A)) and delayed onset muscle soreness (DOMS; Panel (B)) in response to Montmorency cherry (MC) or placebo (PLA) supplementation. * Group effect; $ Interaction effect (p < 0.05).
When data was normalised to baseline values, CMJ also showed significant time (F(1,4) = 30.320, p < 0.001, η2  = 0.684), group (F(1,2) = 7.336, p = 0.017, η2  = 0.345) and interaction (F(1,4) = 3.334, p = 0.028, η2  = 0.193) effects (Figure 3).
 Both MC and PLA groups demonstrated reduced CMJ (vs. baseline) in the 
72 h post-trial period, although the CMJ decrease in the MC group was 
significantly attenuated at 24 h (5%, p = 0.022) and 48 h (6%, p = 0.017) versus placebo. Significant time (F(1,4) = 12.988, p < 0.001, η2  = 0.481) and group (F(1,2) = 7.963, p = 0.015, η2 
 = 0.355) effects were found for the 5-0-5 agility test. MC times for 
the 5-0-5 agility were on average 3% faster (vs. PLA) across the 72 h 
post-trial testing period. For the last of the performance measures, 20 m
 sprint time, significant time (F(1,4) = 9.681, p = 0.001, η2  = 0.409) and interaction (F(1,4) = 3.145, p = 0.035, η2 
 = 0.183) effects were apparent, with both MC and PLA groups 
demonstrating slower times in all three post-trial tests. At 48 h in the
 MC group, 20 m sprint times were significantly (p = 0.043) faster (4%) than PLA. DOMS was significantly increased in both groups across the 72 h post-trial period (F(1,4) = 37.206, p < 0.001). A significant group effect (F(1,2) = 8.486, p = 0.011, η2  = 0.377) showed that DOMS ratings were lower in the MC group (vs. PLA), which were mirrored by interaction effects (F(1,4) = 4.069, p = 0.013, η2  = 0.225) at 24 (p = 0.044), 48 (p = 0.018) and 72 h (p = 0.007), which showed almost complete recovery of at 72 h.

Comparison
 of countermovement jump (CMJ) height with Montmorency cherries (MC) or 
placebo (PLA) supplementation. * Group effect; $ Interaction effect (p < 0.05).
With regards to inflammatory markers, IL-6 (Figure 4) was found to be elevated in both groups following the trial (F(1,8) = 52.180, p < 0.001, η2  = 0.788). Group comparisons (F(1,2) = 10.223, p = 0.006, η2 
 = 0.422) demonstrated an overall significantly attenuated IL-6 response
 to the trial in MC (vs. PLA), with significant interaction effects (F(1,8) = 3.313, p = 0.003, η2  = 0.191) showing peak differences of 3.10 pg·mL−1 occurring immediately post-exercise (p = 0.03). Further inflammatory marker data for plasma IL-8 (F(1,8) = 4.905, p = 0.010, η2  = 0.259), TNF-α (F(1,8) = 6.343, p < 0.001, η2  = 0.312) and hsCRP (F(1,8) = 20.298, p < 0.001, η2 
 = 0.592) revealed significant increases in each variable in the 72 h 
following the trial, however, group and interaction comparisons failed 
to identify differences. IL-1-β was not found to be increased at any 
measurement point across the trial period. The trial significantly 
increased CK in both groups (F(1,4) = 10.243, p = 0.004, η2  = 0.423), although no group or interaction effects were found. Peaks values of 1200 IU/L were attained at 24 h (Table 1).

Interleukin-6
 (IL-6) responses in the Montmorency cherry (MC) and placebo (PLA) 
groups to the adapted Loughborough Intermittent Shuttle Test (LISTADAPT) exercise. * Group effect; $ Interaction effect (p < 0.05).
Lipid hydroperoxides were increased in the 72 h post-exercise period as indicated by a significant time effect (F(1,8) = 5.973, p < 0.001, η2 
 = 0.289). Peak increases of 35% above baseline occurred at 5 h. 
Although there was a tendency for higher PLA group values, no 
significant group or interaction effects were found. A summary of 
variables is reported in Table 1.
In order to identify any group differences in LISTADAPT performance, a comparison of sprint times during the LISTADAPT protocol was performed using Student’s T-test. No group differences in LISTADAPT sprint performance were found (t = 1.511, p = 0.153).
4. Discussion
The
 main finding of this study was that participants supplemented with MC 
were able to maintain greater functional performance than PLA 
counterparts following prolonged intermittent sprint activity. More 
specifically, MVIC, CMJ, 20 m sprint and 5-0-5 agility performances were
 superior in the 72 h post-exercise with MC (vs. PLA). In addition, DOMS
 and IL-6 were lower in the MC group throughout the post-trial period. 
The
 attenuated declines in muscle performance are consistent with the 
findings of previous studies investigating MC as a recovery aid [12,13,14,15,16] and additionally the magnitude of MVIC decline following the LISTADAPT was comparative to previous work utilising a similar protocol [29].
 The MVIC performance was on average 17%, superior in the MC group (vs. 
PLA). MC supplementation also resulted in better (vs. PLA) CMJ 
performance at 24 and 48 h. Sprint times in both groups were slower in 
the 72 h post-exercise period, however at 48 h the MC group was 
significantly faster. Agility (5-0-5) times were also faster in the MC 
group by an average of 3% across the 72 h post-trial period. 
Interestingly, the MVIC results in the 72 h following exercise suggests 
MC supplementation abolishes declines in this performance measure—a 
result that has been previously reported [13].
These
 data support the idea that supplementation with MC protect declines in 
muscle function following strenuous exercise, specifically in activity 
akin to repeated sprint sports and games play such as rugby, soccer and 
field hockey. A reduction in post-exercise IL-6 suggests a lower acute 
inflammatory response to the exercise bout that might contribute to the 
performance differences between groups. The COX, prostaglandin, IL-6 
pathway, which are activated during the secondary inflammatory response 
to cellular disruption, has been associated with proteolytic and 
lipolytic processes [30]
 and subsequently muscular performance could be inhibited. Seemingly, MC
 supplementation reduced (but did not abolish) this process and allowed 
for greater maintenance of muscular performance in the recovery period. 
Conversely, there were no group differences in hsCRP. This is unexpected
 given that IL-6 is implicated as a signalling molecule for the 
expression of hsCRP [31,32].
 We are unable to resolve the discrepancy between IL-6 and hsCRP and 
therefore suggest further work is needed to identify the mechanism by 
which MC might exert its anti-inflammatory responses in response to 
strenuous exercise.
In contrast to previous work [12]
 that examined repeated days cycling exercise, there were no differences
 in LOOH between groups. The obvious discrepancy between study findings 
may be attributed to single versus repeated days exercise. Unlike Bell 
et al. [12],
 where repeated days cycling exercise (metabolic challenge) were used, 
the current study investigated a single bout of strenuous exercise that 
incorporated both a metabolic and mechanical exercise stress. 
Conceptually, the accumulated stress response from repeated day’s 
exercise would be greater than a single bout, but of course the 
modalities, cycling versus simulated concurrent exercise, pose very 
different exercise challenges and hence the redox response may also 
differ considerably between exercise stimuli. 
In agreement with previous work [16,22],
 the lower post-exercise DOMS in the MC group provide further evidence 
for the protective effect of cherries. Despite this, previous work from 
our lab [13]
 using MC has not shown this positive effect. This discrepancy may be 
attributed to the different modes of exercise employed to induce stress.
 Whilst muscle actions during cycling are almost exclusively concentric [33], the repeated sprints and decelerations during the LISTADAPT
 protocol in the present study, place a heavy eccentric load on the same
 muscle groups and as a result are likely to incur greater mechanical 
stress. Indeed, DOMS ratings from the present study were consistently 
higher than those in the cycling studies [12,13].
 In further support of this supposition, CK (an index of cellular 
disruption following damaging exercise) was considerably higher than the
 aforementioned cycling studies. However, the CK response reported in 
previous work [14,15] using protocols that also incorporate a heavy eccentric component, showed no evidence for a protective effect of MC.
The
 high-intensity, prolonged, intermittent nature of soccer and other 
repeated sprint sports places a high degree of both mechanical and 
metabolic stress [2],
 which is reflected by the increase in the appearance of physiological 
stress responses in the present study. Whilst this is not the first 
study to demonstrate accelerated recovery of functional performance with
 MC supplementation, it is the first to do so following simulated games 
play and therefore represents an important wider application of this 
intervention to aid exercise recovery in sports of an intermittent 
sprint nature, such as soccer, rugby, field hockey and basketball. 
Collectively, there is a growing body of evidence suggesting that MC has
 the ability to facilitate exercise recovery—perhaps by modulating 
inflammation and/or oxidative stress. The exact mechanisms behind these 
promising data are not clear, so it seems prudent to explore further; 
perhaps by using animal, cellular and molecular techniques to provide a 
greater understanding of the application of MC and other phenolic-rich 
foods.
In summary, this study provides
 further evidence for the use of MC as a recovery aid. For the first 
time, MC supplementation has been shown to accelerate the recovery of a 
number of functional performance measures following prolonged 
intermittent sprint activity and suggest that some dampening of the 
post-exercise inflammatory processes might be responsible. With regards 
to application, the dampening of such responses could be highly 
advantageous in sports requiring athletes to complete high volumes of 
training whilst ‘in-season’, or athletes competing in tournament 
scenario’s that require multiple performances within a short time period
 and the ability to recover in sufficient time is a challenge. 
Additionally, although dietary restrictions were imposed throughout the 
study period, the results suggest that sports requiring sprinting or 
high intensity directional changes might benefit from MC supplementation
 when playing schedules are congested and recovery time is limited 
between games. Finally, the issue of modulating the post-exercise 
oxidative stress and inflammatory response has raised concerns; insofar 
as these stressors are implicated in the adaptive response. Although 
there is no evidence that the adaptive response is affected by 
functional foods, this question should be addressed in order to 
determine if periodiation of these sorts of supplements is warranted.
Author Contributions
P.G.B.,
 E.S. and G.H. conceived and designed the experiments; P.G.B. performed 
the experiments; P.G.B., G.W.D. and G.H. analyzed the data; G.W.D. 
contributed reagents/materials/analysis tools; P.G.B., E.S. and G.H. 
wrote the paper.
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