THEEFFECTSOF10-DAYSUPPLEMENTATIONOF300MGOFSAFFRONON

DELAYEDONSETMUSCLESORENESSFOLLOWINGECCENTRICEXERCISE

by

BlairWark

BachelorofScienceinKinesiology,UniversityofNewBrunswick,2013

AThesisSubmittedinPartialFulfillmentoftheRequirementsfortheDegreeof

MasterofScienceinExerciseandSportScience

intheGraduateAcademicUnitofKinesiology

Supervisor: UshaKuruganti,PhD,FacultyofKinesiology

ExaminingBoard: J.Noble,PhD.,Chair K.Seaman,PhD.,FacultyofKinesiology

K.Barclay,DepartmentofBiology

ThisthesisisacceptedbytheDeanofGraduateStudies

THEUNIVERSITYOFNEWBRUNSWICK

October,2018

©BlairWark2019

ii

ABSTRACT

Thepurposeofthisstudywastoevaluatetheeffectsof10-day

supplementationwith300mgofsaffrononDelayedOnsetMuscleSoreness

(DOMS).Inapseudo-randomdoubleblindrepeatedmeasurescrossover

counterbalanceresearchdesign,onday7ofsupplementation12maleand5

femaleparticipantscompletedsixsetsof10repsofmaximaleccentric

isokineticcontractions.KneeRangeofMotion(ROM),peakisokinetictorque,

andpainmeasuredviaaLikertscalewererecordedpreand24,48,and72

hourspostexerciseforcomparison.Therewasnosignificantdifference

detectedineitherpeaktorqueorROMmeasuresintheexperimentalvs

control(timextreatment),howevermedianpainscoreswerestatistically

differentbetweenfemaleexperimental(0)vscontrol(1.8)groupat72hours

postexercise,p=.043.Thisstudywasoneoffewtoexaminesaffron

supplementationandDOMSandtoourknowledgethefirsttoincludefemales.

Althoughwehadalimitedsample,sizewedidfindsomepreliminaryevidence

that10-daysaffronsupplementationmaybebeneficialtoreducemuscle

sorenessfollowingeccentricexerciseinuntrainedfemaleparticipants.

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TableofContents

ABSTRACT.....................................................................................................ii

TableofContents.........................................................................................iii

ListofTables................................................................................................vi

ListofFigures..............................................................................................vii

ListofSymbols,NomenclatureorAbbreviations..........................................ix

1.0Introduction............................................................................................1

1.1Purpose..........................................................................................................8

1.2Hypotheses.....................................................................................................8

1.3ImpactofStudy............................................................................................10

2.0LiteratureReview..................................................................................11

2.1.1TheCauseofDOMS...............................................................................11

2.1.2TheoriesRegardingStrengthLossfromDOMS.......................................14

2.2ExerciseType,Intensity,andDurationinrelationtoDOMS..........................15

2.2.1TrainingEffectinrelationtoDOMS........................................................17

2.2.2LimbDominanceandCross-EducationofStrength.................................19

2.2.3TheMagnitudeofDOMSinrelationtothedegreeofmuscledamage....19

2.3TreatmentsresearchedformanagingDOMSandrelatedsymptoms.........21

2.3.1Ultrasound............................................................................................21

2.3.2ContinuousCompression.......................................................................22

2.3.3Anti-inflammatoryDrugs,andTrainingsAdaption.................................23

iv

2.4NutraceuticalsforthemanagementofDOMS...............................................27

2.5Whyisitimportant?.....................................................................................61

2.6Saffron;whatisitandwhatisinit?..............................................................62

2.6.1TheBioactiveCompoundsinSaffron:Crocin,Crocetin,Picrocrocin,and

Safranal..........................................................................................................65

2.6.2ToxicityandSideEffectsofSaffron........................................................68

2.6.3SaffronQualityandGrading..................................................................69

2.7IsokineticDynamometry...............................................................................69

2.7.1GravitationalEffectsonIsokineticMovements......................................71

2.7.2InertialEffectsonIsokineticMovements...............................................71

2.7.3IsokineticandIsometricMaximumTorqueMeasures............................72

3.0Methods................................................................................................75

3.1Participants..................................................................................................75

3.2SaffronManufacturing..................................................................................77

3.3Instrumentation...........................................................................................78

3.4Procedures...................................................................................................79

3.5.1MaximalIsometricandIsokineticTorqueAssessment...........................80

3.5.2ROMAssessment...................................................................................82

3.5.3MuscleSorenessAssessment.................................................................83

3.6ExerciseProtocol..........................................................................................83

3.7DataAnalysis................................................................................................84

4.0Results...................................................................................................86

4.1IsokineticTorque..........................................................................................86

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4.2RangeofMotion...........................................................................................91

4.3PerceivedMuscleSoreness...........................................................................95

5.0DiscussionandConclusion..................................................................100

5.1AnticipatedLimitations...............................................................................105

5.2UnanticipatedLimitations...........................................................................109

5.3Conclusion..................................................................................................110

References................................................................................................112

AppendixA–InvitationLetterandInformedConsent...............................137

AppendixB–WaterlooFootednessQuestionnaire-Revised......................149

AppendixC–WitnessTreatmentForm......................................................150

AppendixD–MaximalIsometricTorqueDataMeasures...........................151

CurriculumVitae……………………………………………………………………………………………

vi

ListofTables

Table2.4.1Comparisonbetweenthecurrentpublishedworksonsaffronand

thepresentresearch.................................................................................39

Table2.4.2Summarizestheresearchonthedifferentnutraceuticalsandtheir

effectsondelayedonsetmusclesorenessandrelatedmeasures............40

Table3.1Pseudo-RandomCounterbalanceResearchDesign..........................76

Table3.2Theindependentanddependentvariablesofinterest.....................79

Table4.1–DescriptiveTorqueData-displaysmeansandstandarddeviation

formaleandfemaletorquedatafortheexperimentaltreatmentandthe

placeboatbaseline,24,48and72hourspostexercise............................88

Table4.2KneeROMDescriptiveData--displaysmeansandstandard

deviationforgenderandchangesinkneeROMdatafortheexperimental

treatmentandtheplaceboatbaseline,24,48and72hourspost

exercise.....................................................................................................92

Table4.3ReportedMuscleSorenessDescriptiveData-displaysmediansand

standarddeviationforgenderandchangesinmusclesorenessratingsfor

theexperimentaltreatmentandtheplaceboatbaseline,24,48and72

hourspostexercise...................................................................................96

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ListofFigures

Figure3.1TestingProtocol.CybexHUMACNorm(CSMI,USAInc.)isokinetic

dynamometerusedtostimulateexerciseinducedmuscledamageandfor

pre-andpost-maximalisometricandisokineticforcemeasures..............78

Figure4.1.1–TorqueDataDistribution-Showstorquedatadistributionfor

thetreatment(Saffronintervention)andcontrol(Placebo)atbaseline,24,

48,and72hourspostexerciseforbothmale(N=10)andfemale

participants(N=5)......................................................................................87

Figure4.1.3–PeakIsokineticTorque-Illustratespeakisokinetictorque(Nm),

overtimeformalesandfemalesandtreatments(experimentalgroupvs

control).Isokineticvelocitywassettonotexceed60degrees/sec.(mean

±SD).........................................................................................................89

Figure4.2.1-RangeofMotionDataDistribution-ShowsROMdata

distributionfortreatment(Saffronintervention)andcontrol(placebo)at

baseline,24,48,and72hourspostexerciseforbothmale(N=12)and

femaleparticipants(N=5)..........................................................................91

Figure4.2.3.RangeofMotion-Rangeofmotionchangesovertimebetween

gender(male/female)andtheexperimentaltreatment/control(mean±

SD).............................................................................................................93

Figure4.3.1ReportedMuscleSorenessDataDistribution-Showsdata

distributionofreportedmusclepainfortreatment(Saffronintervention)

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andcontrol(placebo)atbaseline,24,48,and72hourspostexercisefor

bothmale(N=12)andfemaleparticipants(N=5).....................................95

Figure4.3.2.PerceivedMuscleSoreness-Comparisonbetweenself-reported

painviaavisualpainscale(1-10)ofperceivedpaininchangesovertime

betweengender(male/femaleandtreatments(saffronandcontrol)

(median±SD)...........................................................................................97

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ListofSymbols,NomenclatureorAbbreviations

DOMS:DelayedOnsetMuscleSoreness

CK:CreatineKinaseenzyme

LDH:Lactatedehydrogenaseenzyme

SR:Sarcoplasmicreticulum

ROM:RangeofMotion

NSAIDS:NoneSteroidAnti-inflammatoryDrugs

ISO:InternationalOrganizationforStandardization

Mb:Myoglobin

BDI:Bleomycin-DetectableIron

SOD:Superoxidedismutase

GPX:Selenium-DependentGlutathionePeroxidase

PC:ProteinCarbonyl

MRI:MagneticResonanceImaging

FSR:FractionalSynthesisRate

VO2max:MaximalOxygenConsumption

TAC:TotalAntioxidantCapacity

ALD:Aldolase

FFA’s:FreeFattyAcids

BM:BodyMass

CHO:Carbohydrate

1

1.0 Introduction

Delayed-onsetmusclesoreness(DOMS)isasymptomofexercise

inducedmuscledamagefromunaccustomedstrenuousexercise,orexerciseof

ahighintensityand/orduration,andnotablyeccentricexercise(Clarkson,

Byrnes,McCormick,Turcotte,&White,1986;Tiidus&Ianuzzo,1983).The

symptomsassociatedwithDOMSarewelldocumentedandincludeimmediate

andprolongedimpairmentofmusclefunction(strengthandpower

development),musclesoreness,lossofrangeofmotionalongthejointangle,

stiffnessandswelling,andplasmacreatinekinase(CK)andlactate

dehydrogenase(LDH)enzymespillageintoplasmafromdamagedmuscle

tissue(Cheung,Hume,&Maxwell,2003).Hough,(1902)firstpresentedthe

ideathatthesymptomsassociatedwithDOMSwereduetomuscleinjury,

whichhedescribedasaruptureinthemusclefibersand/ortheconnective

tissueofthetendon.Sincethennumerousworkingtheorieshavesurfacedto

explaintheetiologyofDOMSandrelatedsymptoms.Fromthemounting

evidence,itiswidelyacceptedthateccentricexerciseinducedmuscledamage

isinitiatedbymechanicalstressleadingtomuscleand/orconnectivetissue

damage,andsubsequentinflammatoryresponsesand/orresulting

intramuscularcalciumdisturbancesthatinturnaffectperformanceandpain

sensation(Cheungetal.,2003).DOMShashadmuchattentionoverthelast

fewdecadeswithrespecttowhatcausesit,howitoccurs,andwhatcanbe

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employedtotreatand/orpreventthesorenessandsymptoms.Basedonthe

proposedmechanismsattributedtoDOMS,researchershaveinvestigated

numeroustreatmentstrategies(bothprophylacticandtherapeutic)focused

onalleviatingthesymptoms,promotingrecoveryandultimatelyimprovingthe

consequentialperformancedecrements.Sometreatmentstrategies

investigatedincludecryotherapy(Gulick&Kimura,1996),stretching(Wessel&

Wan,1994),anti-inflammatorydrugs(Donnelly,Maughan,&Whiting,1990;

Gulick&Kimura,1996;Hassonetal.,1993;Stone,Merrick,Ingersoll,&

Edwards,2002),ultrasound(Hasson,Mundorf,Barnes,Williams,&Fujii,1990),

ultrasound&phonophoresis(Ciccone,Leggin,&Callamaro,1991),electrical

currenttechniques(Allen,Mattacola,&Perrin,1999;Butterfieldetal.,1997;

Denegar,Yoho,Borowicz,&Bifulco,1992),homeopathy(Gulick&Kimura,

1996;Vickers,Fisher,Smith,Wyllie,&Lewith,1997),massage(Gulick&

Kimura,1996),compression(Kraemeretal.,2001),hyperbaricoxygentherapy

(Harrisonetal.,2001)andexercise(Armstrong,1984).Ofthesetherapeutic

treatments:onlyultrasound,compression,exercise,andanti-inflammatory

drugshaveshowedpositiveresultsintheimprovementofDOMSandrelated

symptoms(Cheungetal.,2003).Oneofthelatestareasofinterestinthe

preventionandameliorationofDOMSandrelatedsymptomsistheareaof

dietarysupplementationofcertainnutraceuticals.Thetermnutraceuticalisa

hybridofthewords‘nutrient’and‘pharmaceutical’andgenerallyarereferred

3

toasdietarysupplementsderivedfromfoodsourceswhereasthehealth

benefitsarebeyondthoseobtainedfromanormaldiet(Gupta&Gupta,2016).

Overthelast25years,afewnutritionalsupplementshavebeeninvestigated

inregardstotheirroleinthemanagementandpreventionofDOMSand

relatedsymptoms;vitaminsCandEthemostheavilyinvestigated(Averyetal.,

2003;Bloomer,2004;Childs,Jacobs,Kaminski,Halliwell,&Leeuwenburgh,

2001;McBride,Kraemer,Triplett-McBride,&Sebastianelli,1998),andthe

researchfindingsareinconsistent.Thesediscrepanciesmaybeinpartdueto

inconsistencyamongthemusclegroupstested,themagnitudeofmuscle

damageofdifferentexerciseprotocols,mode,intensity,anddurationofthe

exercisetestmeasures,differentdosagesandtimeframeofingestionof

treatments(therapeuticvs.prevention),andthedifferencesofpopulations

investigate(e.g.,age,gender,trainedvsuntrained.Themajorityof

nutraceuticalsexploredincludeandarenotlimitedto:saffron(Meamarbashi

&Rajabi,2015),astaxanthin(Bloomeretal.,2005),fishoilwithflavonoids

(Lennetal.,2002),creatine(Rawson,Gunn,&Clarkson,2001),HMBanda-

ketoisocaproicacid(VanSomeren,Edwards,&Howatson,2005),L-carnitineL-

tartrate(Voleketal.,2002),chondroitinsulfate(Braun,Flynn,Armstrong,&

Jacks,2005),proteaseenzyme(Bailey,Barnes,Derr,Hall,&Miller,2004;Beck

etal.,2007),pomegranatejuice(Trombold,Reinfeld,Casler,&Coyle,2011),

tartcherryjuice(Connolly,McHugh,&Padilla-Zakour,2006),bromelain(Stone

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etal.,2002),ginger(Matsumura,Zavorsky,&Smoliga,2015),branched-chain

aminoacids(Shimomuraetal.,2010),proteins(Etheridge,Philp,&Watt,

2008;Nosaka,Sacco,&Mawatari,2006),andcarbohydrates(Luden,Saunders,

&Todd,2007;Whiteetal.,2008).Sofar,agoodmajorityofthese

supplementshaveshowntohavenoeffectonpreventingDOMS,butsome

haveshownpromisewithreportedfavorableresultsalleviatingsome,butnot

allofthesymptoms(Connolly,McHugh,etal.,2006;Etheridgeetal.,2008;

Machinetal.,2014;Matsumuraetal.,2015;Shimomuraetal.,2010;Bakhtiar

Tartibian,Maleki,&Abbasi,2009;BakhtyarTartibian,Maleki,&Abbasi,2011;

Trombold,Barnes,Critchley,&Coyle,2010;Tromboldetal.,2011).

Saffron,awidelyavailablecookingspiceisoneofthemostrecentof

thesedietarysupplementsunderinvestigation.Ofthenutraceuticals

investigatedthusfar,saffronhasexhibitedsomepotentialtopreventDOMS

andrelatedsymptoms(Meamarbashi&Rajabi,2015).Recently,Meamarbashi

andRajabi(2015)investigatedthepreventativeeffectsofsaffronand

indomethacinsupplementationonbiochemicalandfunctionalindicatorsof

DOMSafter1-sessionofunaccustomedeccentricexercise.Themost

interestingfindingwasthatthesaffrontreatmentgroupexhibitedsignificant

increasesinisometricforceoutputabovebaselinemeasures24,48,andtoa

largerdegree72hoursfollowingonetaxingexercise.Surprisingly,thesaffron

treatmentgroupreacheda63.3%increaseinmaximalisometricforceoutput

5

fromtheirbaselinemeasures72hourspostexercise.Moreover,thecontrol

groupsignificantlydecreasedmaximalisometricforcedevelopmentby24.3%

andby72hourspostexercise.Thesaffroninterventionalsoshowedno

declineinisotonicforcedevelopmentandonlysomereportedmusclepain24

hours’post-exercise.Thecontrolgroupdidnotreachfullyrecoverofmuscle

painorisotonicforcedevelopmentby72hours’post-exercisewhilethe

indomethacingroupdidattainreliefofmusclesorenessby72hours’post-

exercise.Theseimpressiveresultsmayhavebeenduetoastrongpreventative

effectwithsaffrononDOMS,howeverthesaffrontreatmentgroupnot

achievingatruebaselinemaybemorelikely.Consideringtheweightloadused

forthelegpresstoinduceDOMSwassetto80%oftheirmaximumisotonic

force(baseline),itispossiblethat,ifthebaselinewerenottrulymaximal,then

theexerciseintensityselectedtoinduceDOMSwouldhavebeenlessthan

intended.Theparticipantsrecruitedwerealsosedentarywithlittletono

previousexperienceusingalegpressmachine.Becauseitisunlikelyoreven

theoreticallypossibletoachievesuchalargeincreaseinforce,production

followingarepeatedtrainingload.Itmaybemorelikelythatalearningeffect,

orbyalackofattainingatruebaselinecouldexplaintheseresultsratherthan

someperformanceenhancementeffectofsaffron.Theauthorsalsoconcluded

thatsaffronsupplementationof300mgadayfor7dayspriortostrenuous

eccentricexerciseappearstobemoreeffectiveatpreventingDOMSand

6

relatedsymptomsthentheleading75mgtherapeutictreatmentof

indomethacinsupplementationinnon-activeyounguniversitystudents.

Somesupportingevidenceexistsonthepotentialofsaffrontoprevent

muscledamageand/orimprovemusclerecoveryfollowingtrauma.Saffron

supplementationhaspreviouslybeenshowntohaveaprotectiveeffecton

cellularmembraneintegrity,suchasratspermcellularintegrity(Vaez,

Mardani,&Razavi,2014)andredbloodcellmembraneintegrity

(Meamarbashi&Rajabi,2013).Saffronalsocontainsanti-oxidant,anti-

inflammatory,andanti-nociceptivecompounds(Hosseinzadeh&Younesi,

2002).Moreover,oneofthefourmainbioactivecomponentsofsaffron

(crocetin)hasbeenrecentlyshowntoimprovewhole-bodyoxygen

consumptionandincreasetherelativegrowthofnormalratmusclecells

(Wilkins,Gainer,&Wilkins,1977).Becausesaffronshowspotentialtoprevent

muscledamageand/orimprovemusclefunctionfollowingstrenuousexercise,

furtherstudiesarecalledfortofirstverifythereliabilityandvalidityofthe

previousfindingsreportedbyMeamarbashiin2015.Replicationofthis

researchinadifferentlab,onadifferentpopulation,withadifferentexercise

protocol,withadouble-blindcrossoverrepeatedmeasuresdesign,wouldhelp

addresssomeofthelimitationsarisingfromthepreviouspublishedfindings.

Moreover,ifoutcomes/resultsaresimilarthenfurtherstudiesshouldbe

conductedtouncoveranypotentialperformanceenhancementbenefitsof

7

thisspice(ifany),themechanismofaction(howitworks),andbecauseitis

expensive,theminimumdosagerequiredtoobtainthedesiredeffect.

However,emergingresearchsuggeststhatinflammationprocesses

maybeessentialfortrainingadaptionstooccur,andblockingtheseprocesses

maynotbeidealforathletesconcernedwithmaximizingmusculartraining

adaptions(Schoenfeld,2012;Tscholl,Gard,&Schindler,2016).However

otherresearchindicatesapositiveeffectinthoseuntrainedwiththeuseof

nonesteroidanti-inflammatorydrugs(NSAIDS)towardsreducingmuscle

sorenessandimprovingperformancewithouthinderingadaptiveprocesses

(Paulsenetal.,2010).Moreoverotherresearchindicatesthatthosebetween

theagesof60-85showasignificantgreaterincreaseinmusclehypertrophy

andstrengthgainswhensupplementedastandardoverthecounterdose

(1200mg)ofibuprofen(Petersen,Beyer,etal.,2011).Moreoverother

researchersreportnodifferencesinmusclehypertrophywhensupplemented

thesamestandarddosefollowinga12weektrainingregimeformenand

womenbetweenagesof50-70(Petersen,Beyer,etal.,2011).However,

consumptionofNSAIDSoverthis12-weekstudydidresultingreaterstrength

gainsinmaximalisometricstrength,maximaleccentricstrengthandeccentric

worksascomparedtotheplacebo.Therefore,itmayonlybeadvantageousfor

theoccasionaluseofsuchmodalitiesforathletesatcompetitionswhere

expressingone’sfullpotentialunhinderedbyDOMS.Conversely,forthe

8

elderly,untrainedorthosedealingwithchronicinflammationtheremaybea

benefitwiththeuseofsuchanti-inflammatorymodalitiesduringtraining.

Thisresearchisvaluableinformationforthoseconcernedabout

musculardiscomfortandpainthatcanbeassociatedwithsports,training,and

strenuousactivity.Moreover,themagnitudeorintensityoftheexercise

appearstoberelatedtothedegreeofmuscledamage,butnotmuscle

soreness(Nosakaetal.,2006).Consideringthatmusclesorenessappearsto

recoverfasterthanmusclefunction(Cleak&Eston,1992),iftrainingis

resumedbeforefullmusclefunction,thereductioninconsequential

performancemaylimittrainingadaptationsduetotrainingatalower

functionalcapacity.

1.1Purpose

Thepurposeofthisresearchistoinvestigatethepreventative

effectivenessof10-daysupplementationwith300mgofsaffronondelayed

onsetmusclesorenessandselectedrelatedsymptoms.

1.2Hypotheses

Ho–Thattherewillbenosignificantdifferencebetweentheplaceboand

treatmentgroupsself-reportedmusclesorenesspre-and24,48,and72hours

postexercise.

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H1–Thattherewillbeasignificantdifferencefoundbetweentheplaceboand

treatmentgroupsself-reportedmusclesorenesspre-and24,48,and72hours

postexercise.

Ho–Thattherewillbenosignificantdifferencebetweentheplaceboand

treatmentgroupskneerangeofmotionpre-and24,48,and72hourspost

exercise.

H1–Thattherewillbeasignificantdifferencefoundbetweentheplaceboand

treatmentgroup’skneerangeofmotionpre-and24,48,and72hourspost

exercise.

Ho–Thattherewillbenosignificantdifferencebetweentheplaceboand

treatmentgroupsisometricpeaktorqueatajointangleof50degreespre-and

24,48,and72hourspostexercise.

H1-Thattherewillbeasignificantdifferencefoundbetweentheplaceboand

treatmentgroups’isometricpeaktorqueatajointangleof50degreespre-and

24,48,and72hourspostexercise.

Ho–Thattherewillbenosignificantdifferencebetweentheplaceboand

treatmentgrouppeakisokinetictorqueataconstantspeedof60degreesper

secondpre-and24,48,and72hourspostexercise.

H1–Thattherewillbeasignificantdifferencebetweentheplaceboand

treatmentgrouppeakisokinetictorqueataconstantspeedof60degreesper

secondpre-and24,48,and72hourspostexercise.

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1.3ImpactofStudy

Thisstudyinvestigatedthepotentialforsaffrontopreventand/or

manageDOMSandrelatedsymptomsfollowingunaccustomedstrenuous

eccentricexercise.Ifthequalitiesofsaffronpermitcellularprotectionofsome

mannerand/oraspeededrecoveryfromtrauma,itcouldserveasan

invaluablesupplementforconcernedwithmusclesoreness.

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2.0 LiteratureReview

Thefollowingchapterwillreviewthecurrenttheoriesregardingthe

causeofDOMSandaccompanyingstrengthloss.Trainingeffectaswellasthe

effectsofdifferentexercisetypesandtheireffectsonDOMS.Basedonthe

currentknowledgeofthetheoriesofthepotentialcausesofDOMS,many

treatmentstrategiesandnutritionalsupplements(nutraceuticals)are

reviewedforthemanagementofDOMS.Thischapterwillhighlightthe

majorityoftheseinterventionandpreventativestrategiesandmore

importantlytherecentfindingsonsaffronanditsprotectiveeffecttowardsthe

managementofDOMS.

2.1.1TheCauseofDOMS

Delayed-onsetmusclesorenessisasymptomofexerciseinduced

muscledamagefromunaccustomedstrenuousexerciseofhighintensity

and/orduration,andisespeciallyprevalentfollowingeccentricexercise

(Clarksonetal.,1986;Tiidus&Ianuzzo,1983).Thesymptomsassociatedwith

muscledamagearewelldocumentedandinclude;immediateandprolonged

impairmentofmusclefunction(strength),delayedonsetmusclesoreness,

stiffness,swelling,andlossofrangeofmotionalongthejointangle

(Armstrong,1984).Musclesorenessisoftenfeltduringmovementor

palpationoftheaffectedmuscle.DOMSdevelops8to24hoursfollowingmost

notablyunaccustomedeccentricexerciseandpersistsusuallyforupto7days,

12

climaxingwithin1-3dayspostexercise(Ebbeling&Clarkson,1989).

Researcherscommonlyreportaprolongedlossinmusclestrength(of50to

60%)immediatelyfollowingeccentricexercisethatcanlastupto10days

(Clarkson,Nosaka,&Braun,1992).Althoughexerciseinducemuscledamage

(EIMD)usuallycausesDOMSandisoftenusedtoquantifyDOMS,theyarenot

equivalent.TheydonotsharethesametemporalrelationshipandDOMSmay

notaccuratelyreflectthephysiologicalresponseof(EIDM).Sincestrengthloss

isapparentimmediatelyfollowingtaxingexerciseandmusclesorenessisnot

feltuntilsomehourslateranddiminishesbeforemaximalforcedevelopment

isrestored;musclesorenessmaynotbeamajorcontributingfactorforthe

lossofforcedevelopment.However,peakedemalevelsmeasuredvialimb

girthhasbeenshowntosharethesametemporalsequenceasdelayedmuscle

soreness(Gulick&Kimura,1996)

Hough,(1902)wasthefirsttosuggestthatthecauseandassociated

symptomsassociatedwithDOMSwasindeedduetomuscleinjury,whichhe

describedasaruptureinmusclefibersand/ortheconnectivetissueofthe

tendon.SincethentheetiologyofDOMShasbeenwellresearchedgivingrise

tofiveotherpopularproposedtheoriesincluding:thelacticacidtheory,

musclespasmtheory,connectivetissuetheory,inflammationtheory,andthe

enzymeeffluxtheory(Cleak&Eston,1992;Gulick&Kimura,1996).Thelactic

acidtheoryhasbeenlargelyrejectedaseccentricexercisehasshownto

13

producelesslacticacidintheworkingmusclesthenconcentricexercise

(Clarkson,1995).Furthermore,(Asmussen,1956)concludedafteraseriesof

experimentsthatmuscularsorenessismorelikelyduetomechanicalstress

andnotbymetabolicwasteproducts.Themusclespasmtheoryisloosely

basedonthepremisethatlocalizedmotorunitspasmsmayleadto

compressionofbloodvesselsleadingtoischemiaandaccumulationofpain

substances.Overthelast100yearsviahistologicalandultra-structural

examinationmountingevidencesupportsHough’sclaimofmuscledamagevia

reporteddamagetothefollowingmuscularcomponents:T-tubules,myofibrils,

thesarcolemma,andthecytoskeleton(Armstrong,Ogilvie,&Schwane,1983;

Friden,Sjöström,&Ekblom,1983).Z-discimpairmentisalsoacommon

characteristicofexerciseinducedmuscledamageandappearstoworsen24-

72hourspostexerciseandoftentakesupto3weekspostexercisetofully

recover(Jones,1996;Newham,McPhail,Mills,&Edwards,1983).Inaddition,

Morgan,(1990)hypothesizedthatsomesarcomeresmay“pop”fromthe

strainorstressofalengtheningmuscle.SupportingMorgan’stheoryTalbot&

Morgan,(1996)reportedrandomlyscatteredsarcomeredisorientationina

stretchedmuscle.Inadditiontomuscledamage,connectivetissuedamage

maybeacontributingfactorasincreasedcollagenbreakdownisalsoobserved

daysaftereccentricexercise(Brown,Child,Day,&Donnelly,1997).The

inflammationtheoryisbasedonthepremisethatinflammatoryresponses

14

mayinitiatefurtherdegradationofproteinstructuresfollowinginitialinjury,

whichultimatelyincreasesosmoticpressuresthatmaytriggerpainreceptors

(Smith,1991).Lastly,theenzymeeffluxtheoryisbasedontheassumption

thatcalcium,whichisnormallyfoundinthesarcoplasmicreticulum,

accumulatesinthedamagedmuscletissuefollowingsarcolemmadamage

(Armstrong,1984).Fromthemountingevidence,itiswidelyacceptedthat

eccentricexerciseinducedmuscledamageisinitiatedbymechanicalstress

leadingtomuscleand/orconnectivetissuedamage,andsubsequent

inflammatoryresponsesand/orresultingintramuscularcalciumdisturbances

thatinturneffectperformanceandpainsensation(Cheungetal.,2003).

2.1.2TheoriesRegardingStrengthLossfromDOMS

Muscledamageviaunaccustomedstrenuousexercisehasbeenshown

tosignificantlyreduceforcedevelopmentcapacityofaffectedmusclesduring

maximalvoluntarycontractions(Komi&Buskirk,1972)andwhenelectrically

stimulated(Davies&White,1981).Afewtheorieshavebeenproposedabout

thepotentialunderlyingmechanismscontributingtothelossofforce

developmentfollowingeccentricexerciseinducedmuscledamage.For

instance,(Ingalls,Warren,Williams,Ward,&Armstrong,1998)suggestedthat

forcelossesimmediatelyfollowingmuscleinjurymightbeduetodecreased

contractileproteinsand/orproblemswiththeexcitation-contractioncoupling

process.In1993,researchersfoundthatadministeringcaffeine,whichwas

15

thoughttoenhancecalciumreleasefromthesarcoplasmicreticulum(SR),

reducedtheconsequentialforcedecrementsfollowingexerciseinduced

muscledamageinrats(Warrenetal.,1993).Thiswouldsuggestthattheforce

decrementsfollowingunaccustomedeccentricexercisemightbedueto

reducedcalciuminflux,whichwouldlimittheexcitation-contractioncoupling

process.Furthermore,Ingalls,(1998)reportedthatexcitation-contraction

failurecouldexplainupto75%oftheforcedeclinesfrom0to5dayspost

exerciseinmice.Thissuggeststhatdisturbancesinthefunctionofthe

sarcoplasmicreticulummayberesponsiblefortheconsequentialforce

decrementsobservedfollowingunaccustomedeccentricexercise.Moreover,

recentstudiesshowevidencethatcaffeineaffectsisometricmaximalforce

andenduranceforthelowerbodymusculature,buttheuppermusculature

remainsunclear(Davis&Green,2009).

2.2ExerciseType,Intensity,andDurationinrelationtoDOMS

Thedegreeofmuscledamageappearstoberelatedtothetypeof

exercise(Clarksonetal.,1986;Talag,1973),thedurationandintensity(Tiidus

&Ianuzzo,1983).In1986,researcherscomparedequivalentconcentric,

isometric,andeccentricmuscularworkprotocolswithsubjectivepainratings

aswellasplasmaserumcreatinekinase(CK)concentrations(Clarksonetal.,

1986).TheyreportedthatdespitesimilarincreasesinCKlevelsfollowingthe

threeregimens,themagnitudeoftheperceivedmusclesorenesswashighest

16

fortheeccentricexerciseregimen.EarlierTalag(1973),comparedexhaustive

concentric,eccentricandstaticcontractionsonindicesofmuscularpain,

function,andlimbvolume.Similarly,Talag,(1973)alsoreportedhigherpain

ratingsofresidualmuscularsorenessfollowingtheeccentriccontractions

ratherthantheconcentricorstaticgroups.Furthermore,musclestrength

remaineddepressedintheeccentricregimenandexhibitedatemporal

relationshipwithmusclesorenessthatwasnotobservedintheothergroups.

Moreover,downhillrunning,whichincorporatesproportionallymoreeccentric

muscularcontraction,hasbeenfoundtoinducemoremuscledamagethan

leveloruphillrunning(Eston,Mickleborough,&Baltzopoulos,1995).Lieber

andFriden,(1993)foundthatthedegreeofmuscleinjuryviaeccentric

exerciseismorepronouncedbythechangeinthelengthofthemusclerather

thanthemagnitudeofforcegenerated.Inotherwords,highforcesimposed

onthemuscleinpartcontributestothemuscledamage,butthemagnitudeof

thestrain,thelengthofthemuscleengagedratherthantheforceonthe

muscleitself,appearstobetheleadinginfluencecontributingtomuscle

damageandrelatedsymptomsofDOMS.Studieshaveshownthatthe

magnitudeofdamageisgreaterfollowingeccentricexerciseinmusclesofa

longerlengthratherthanshorterlength(Child,Saxton,&Donnelly,1998),

indicatingthatstrainmayindeedbetheleadingcauseofthedamage.

Moreover,eccentricexercisehasshowntoproducelesslacticacidinthe

17

workingmusclesandisthereforelessmetabolicallystressful,soother

mechanismssuchasmechanicalstressmustberesponsibleforthemuscle

damagefollowingeccentricmuscularcontraction(Clarkson,1995).In1983,

researchersinvestigatedtherelationshipbetweenexerciseintensityand

durationonmuscledamageandsorenessbyadjustingthepercent10-

repititionmaximum(%10RM)andthetotalnumberofcontractions(NR)

performed(Tiidus&Ianuzzo,1983).Theyfoundthatincreasingthedurationor

intensityoftheexerciseresultedingreatercorrespondingincreasesinmuscle

damageviaincreasesinserumCK,LDHenzymelevels,andmusclesoreness.

Moreimportantly,theyconcludedthathighintensityshortdurationexercise

boutsresultedinhigherlevelsofmuscledamageandsoreness.

2.2.1TrainingEffectinrelationtoDOMS

Trainingandevenperformanceofasingleboutofexerciseappearsto

notonlyhaveaprotectiveeffecttowardsfurthermuscledamage,butmay

alsoenhancerecoverytosuchdamageinrepeatedevents,providedthereis

anappropriaterecoveryperiod.Byrnes&Clarkson,(1986)reportedthat

followingtwo30-minexerciseboutsofdownhillrunningseparatedby3,6,

and9weekintervalsthatperformanceofasingleexercisebouthada

prophylacticeffectonrepeatedexerciserelateddamageandDOMSand

serumproteinresponsesthatlastedforaslongas6weeks.Clarkson&

Tremblay,(1988)investigatedexercise-inducedmuscledamage,repair,and

18

adaptationfollowingthreeeccentricexercisesinvolvingtheforearmflexors.

Theresearchershadeightwomenperform70maximalcontractionswith1

armand24maximalcontractionswiththeotherarmandrepeatedit2weeks

later.MeasuresofserumCK,soreness,isometricstrengthandmuscle

shorteningwereobtainedthedayoftesting(pre-andpost)andtheneachday

for5daysfollowing.Comparisonofthe70-MAXconditioncriteriameasuresto

the70-MAX2showedsignificantlyimprovementsmoresothanthe24-MAX

conditions.Theyreportedthatinregardstotheabilityofthemuscletoadapt

toexerciseinducedmuscledamagethemuscleismoreresistanttodamage

andanydamagethatmayoccurisrepairedatafasterrate.Similarfindings

frombothmenandwomenperformingcontinuouseccentricelbowflexor

actionshasbeenreportedregardingimprovedrecoveryratesofbothstrength

andforce-frequencycharacteristicsfollowingexerciseinducedmuscledamage

ofthree20-minuteexerciseprotocols,spaced2weeksapart(Newham,Jones,

&Clarkson,1987).Inshort,trainingandevenperformanceofasingleboutof

exercisegiventheappropriaterecoveryperiod(of2weeks)appearstohavea

prophylacticeffectonDOMSandrelatedsymptomsofDOMSthatappearto

persistupto6weeks.Moreimportantly,inadditiontoenhancedresistanceto

exerciseinducedmuscledamagefollowingrepeatedboutsofexercise,the

recoveryofthesymptomsofDOMSappeartobeenhancedonlywithinthis6-

weekperiod.

19

2.2.2LimbDominanceandCross-EducationofStrength

Hageman,Gillaspie,andHill,(1988)evaluatedtheeffectsofspeedand

legdominanceontorquevaluesandratiosforthequadricepsandhamstrings

duringbothconcentricandeccentricexerciseat30and180degrees/sec.They

foundthatduringboththeconcentricandeccentricexercisethatthe

quadricepsalwaysproducedagreatertorquethanthehamstringsatboth

speeds.Inaddition,moreimportantly,theyfound,forthemalesonly,the

hamstrings/quadricepsratiosofthenon-dominantlegweresignificantly

greaterthanthedominantlegforbotheccentricandconcentricexercisesand

speeds.Moreover,nosignificantdifferenceswereobservedintorquevalues

betweenthedominantandnon-dominantlegsduringbothconcentricor

eccentricexerciseandspeeds.Recently,researchershaveprovidedevidence

thatcross-educationofstrengthbetweenthelimbsisgreaterwhen

transferredfromthedominanttothenon-dominantlimb(Farthing,2009).

Therefore,ifcomparisonsbetweenrightandleftlegfunctionalparameters

followingexerciseareemployedcarefulconsiderationshouldbetakentouse

thenon-dominantlimbfirstandthenthedominantlimbtominimizethe

cross-educationeffectofstrengthdevelopment.

2.2.3TheMagnitudeofDOMSinrelationtothedegreeofmuscledamage

Nosaka,Newton,andSacco,(2002)investigatedtherelationship

betweenDOMSandbiochemicalandfunctionalindicatorsofmuscledamage.

20

Themagnitudeofmusclesorenesswasassessedviathreepainvisualanalog

scales:1)palpationsassessmentSOR-Pal)2)flexionassessment(SOR-Flx)3)

andextensionassessment(SOR-Ext).Musclesorenesswasevaluatedwhenthe

elbowflexorswerepalpated(SOR-Pal),Flexed(Sor–Flx)andstretched(Sor-

Ext).Allmeasuresweretakenimmediatelybeforeandaftertheexerciseand

theneachdayforthenextfourconsecutivedays.Themusclesorenessscales

andindirectindicatorsofmuscledamagewerebothcomparedbetweenthe

threedifferentexerciseprotocols.Theexerciseprotocolsinvolved12,24,and

60maximaleccentricactionsofelbowflexorsusingthenon-dominantarmon

110maleuniversitystudents.Theresultsindicatethatthemagnitudeof

muscledamageviatheindirectmeasuresofmuscledamagewerelargerwith

eachincreaseinthemagnitudeoftheexerciseprotocol.Inaddition,ablunted

recoverytimeframewasalsoobservedforboththe24and60maximal

eccentricprotocolconditionsascomparedtothe12maximaleccentric

contractions.Moreimportantly,poorcorrelationswereobtainedbetweentwo

ofthevisualpainscalesusedtoquantifyDOMSandtheindirectmeasures

usedtoquantifytheextentofthedamage.Thequantificationofmuscle

sorenessviapalpationandduringflexionmaynotbeanidealmeasure

consideringthesemarkersreflectedthemagnitudeofthemuscledamage

poorly.Theresultsindicatethatperceivedmusclesorenessishighlysubjective

andmaybedifficulttoquantifyoruseasareliableindicatortoreflectthe

21

extentormagnitudeofthedamageespeciallywhencomparingthelevelof

sorenessbetweentwodifferentgroups,orevenchangesovertimewiththe

samesubjects.However,Sor-Extshowedthemostmeaningfulcorrelation

betweenthethreescalesusedandallindirectmeasuresofDOMSinvestigated

andthereforewouldbethemostappropriatemeasureforfuturestudies.

2.3TreatmentsresearchedformanagingDOMSandrelatedsymptoms

BasedontheproposedmechanismsattributedtoDOMSfromexercise

inducedmuscledamageresearchershaveinvestigatednumeroustreatment

strategies(bothprophylacticandtherapeutic)focusedonalleviating

symptoms,promotingrecoveryand/orimprovingtheconsequential

performancedecrements.Ultrasound,compression,exerciseandanti-

inflammatorydrugsandhaveshowedpositiveresultsintheimprovement

DOMSandrelatedsymptoms(Cheungetal.,2003).

2.3.1Ultrasound

In1990,researcherscomparedtheanalgesiceffectofpulsating

ultrasoundtoaplacebogrouponDOMSfollowinganeccentricexercisebout

(Hassonetal.,1990).Ultrasoundwasadministered24hours’post-exerciseto

thevastuslateralisandmedialismusclesatafrequencyandintensityof1

MHz,and0.8W/cm2.Thisresultedinasignificantreductioninself-reported

soreness48hourspostexerciseascomparedtothecontrol.Althoughthe

22

mechanismresponsibleforthereducedperceivedsorenessfollowing

ultrasoundisunknown,theauthorsattributedthetreatmenteffectto

decreasingintramuscularpressureand/orareducedinflammationresponse.

Incontrast,arelatedstudyinvestigatedtheeffectsofultrasoundand

phonophoresususingatopicalanti-inflammatorycream(trolaminesalicylate)

onDOMSusingafrequencyof1MHzandanintensityof1.5W.cm2and

reportedincreasedmusclesoreness(Cicconeetal.,1991).Phonophoresisisa

methodofdrugadministrationwhereultrasoundwavesareusedtoenhance

thedeliveryofpharmaceuticalagentswithinthetissues.Theyfoundthat

althoughallthegroupsexperiencedsoreness;theultrasoundgroupalone(no

anti-inflammatorycream)experiencedtheonlysignificantincreasein

perceivedsoreness48hourspostexercisewhencomparedtothecontrolarm.

Thus,morestudiesarerequiredtoidentifytheappropriatefrequencyand

intensityfortherapeuticapplicationofultrasoundtherapyinthemanagement

ofDOMSsymptoms.

2.3.2ContinuousCompression

Continuouscompressionhasbeenshowntobeaneffective

therapeuticinterventioninthetreatmentofeccentricexercise-inducedmuscle

soreness(Kraemeretal.,2001).Researchersinvestigatedwhethera

compressionsleevewornimmediatelyaftermaximaleccentricexercisewould

enhancemusclerecovery.Fifteenhealthmenwererandomlyassignedto

23

eitheracontrolgrouporacontinuouscompression-sleevegroup.The

researchersfoundfollowingtwosetsof50armcurlsthatthecompression-

sleevegroupshowedalowerelevationofplasmacreatinekinase(CK)thanthe

control.Inaddition,thecompressionsleevegroupshoweddecreased

perceptionofsoreness,reducedswelling,andimprovedfunctionalrecovery

(forcegeneration)comparedtothecontrol.Thisstudyindicatesthat

compressionviauseofacompressivearmsleevewornpost-exercisemay

reducetheseverityanddurationofthesignsandsymptomsassociatedwith

muscledamage.Morerecently,Hill,Howatson,Someren,Leeder,&Pedlar,

(2013),conductedasystematicreviewof12studiesthatevaluatedthe

efficacyofcompressiongarmentsonmeasuresofDOMS,musclestrength,

powerandcreativekinase(CK).Theyconcludedthattheuseofcompression

garmentshadamoderateeffectinreducingtheseverityofDOMS,muscle

strength,power,andCK.Theseresultsindicatethatcompressiongarments

maybebeneficialforenhancingrecoveryfrommuscledamage.

2.3.3Anti-inflammatoryDrugs,andTrainingsAdaption

Anti-inflammatorieshavebeenshowntoreducetheamountofmuscle

swelling(oedema)andassociatedintramuscularpressures,whicharetwo

contributingfactorstopainandmusclesoreness(Cheungetal.,2003).

Inflammationandthesubsequentedemawhichisvaluedasanintimatepart

oftheadaptationandrecoveryprocessfollowingtissuedamagemayactually

24

contributetothedevelopmentofDOMS,aspeakoedemalevelsappearto

coincidewithmusclesoreness(Armstrong,1984;Gulick&Kimura,1996).

Thus,anumberofanti-inflammatorydrugshavebeeninvestigatedforboth

thepreventionandtherapeutictreatmentofDOMSandrelatedsymptoms.A

prophylacticdosageofIbuprofen(400mg)hasshowntoreducemuscle

sorenessperceptionandseemstoassistinrestoringmusclefunction

significantlymorethantherapeuticibuprofentreatmentof400mg(Hassonet

al.,1990).However,otherresearchersreportedthatoraldosesof1200mgof

oxaprozinimmediatelyfollowingexerciseandagaineachdayfor3dayswas

ineffectiveinabatingthesignsandsymptomsassociatedwithDOMS(Gulick&

Kimura,1996).Moreimportantly,thehigherdoseswerenotonlyineffectiveat

preventingsignsandsymptomsofDOMS,butthetreatmentof1200mgaday

forthe3-dayperiodappearedtoimpedemusclerecoveryandfunction.The

authorsconcludeditispossibleinflammatoryprocessesmaybenecessaryfor

musclerecoveryandhighdosesofanti-inflammatorymedicationmayimpede

theproductionofmyofibrillarproteins.Moreover,largerdosesofibuprofen

havebeenshownincreaseoraggravatethelevelofmusculardamage

followingdownhillrunningandisnotrecommendedasanappropriate

treatmentstrategyforDOMSandrelatedsymptoms(Donnellyetal.,1990).

Lastly,Stone,(2002)foundthatibuprofensupplementationof400mgtaken

threetimesadayfor3daysimmediatelyfollowingexerciseinducedmuscle

25

damagehadnoeffectonelbowflexorpain,ROM,orpeaktorquemeasures.

Consideringtheibuprofengroupreceivedastandarddoseof400mgthree

timesadaytotalinga1200mgdailydose,NSAIDSmaynotbetheideal

treatmentofDOMSandrelatedsymptoms.

WithrespecttothepotentialforNSAIDStolimittrainingadaptions

Krentz,Quest,Farthing,Quest,&Chilibeck(2008)showedthatamoderate

dose(400mgibuprofen)takenoverthecourseofaresistancetraining

regimendidnotimpairoreffectmusclehypertrophy,orstrengthanddidnot

affectratingsofmusclesorenessinyounguntrainedindividuals.Similarly,

Paulsenetal.,(2010)showedapositiveeffectinthoseuntrainedwiththeuse

ofNSAIDStowardsreducingmusclesorenessandimprovedperformance

withouthinderingmuscularadaptation.However,otherpublishedresearch

indicatesadetrimentaleffectoflargerstandardoverthecounterdosesof

ibuprofen(1200mg)onmuscleadaptationforyounghealthytrained

individuals(Trappeetal.,2002),butnottheelderlywithkneeosteoarthritis

betweentheagesof50-70(Petersen,Beyer,etal.,2011;Petersen,Miller,

Hansen,Kjaer,&Holm,2011).Inaddition,otherresearchersreportedthat

evenadoseof45to100mgofindomethacincanlimittrainingadaptionsin

youngtrainedenduranceathletes(Mackeyetal.,2007;Mikkelsenetal.,

2009).Moreover,whenconsumedincombinationwitharesistancetraining

regime,standardoverthecounterdosesofibuprofenhaveshownpositive

26

hypertrophiceffectintheelderlybetweentheagesof60-85whosufferfrom

chronicinflammation(Trappeetal.,2010),butnotthosebetweentheagesof

50-70(Petersen,Miller,etal.,2011).Althoughnoadditionalhypertrophic

responsewasnotedforthosebetweentheagesof50-70,consumptionofthe

standardoverthecounterdoseofibuprofendidresultingreatergainsin

maximalisometricandeccentricstrengthascomparedtotheplacebo.

Thesefindingssuggestthattheelderly,untrainedand/orthose

managingadegreeofchronicinflammationmaybenefitfromanti-

inflammatorieswithoutimpedingmuscleadaptations,atleastintheshort

term.Thiscouldexplainwhystandardoverthecounterdosesofibuprofen

showpositiveeffectsinthispopulation.Itispossiblethatthoseuntrained

haveatendencyto“overshoot”theinflammationresponseandmayexplain

whylowerdosessuchofNSAIDShasshownnodetrimentaleffectonmuscle

adaptationandsomerelieffrommusclesoreness(Burdetal.,2009;Paulsenet

al.,2010).Itseemsclearthatinflammationprocesses,oratleastacertain

amountthereof,arekeytoobtainingmuscularadaptations(Tscholletal.,

2016).However,itiscurrentlyunclearhowmuchinflammationisappropriate

forthesemuscularadaptations,butwhenadvisingNSAIDSandotheranti-

inflammatorymodalities,itisimportanttorememberthatthereisnohealing

withoutinflammation,buttherecanbeinflammationwithouthealing.

Schoenfeld(2012),whoextensivelyevaluatedtheeffectsofNSAIDSonmuscle

27

growthanddevelopment,concludedthattheoccasionaluseofNSAIDSarenot

likelytoaffectmuscledevelopment,howevertheefficiencyfortheirusein

alleviatinginflammatorysymptomsinthismannerremainsquestionable.This

isagrowingconcernforthosemanagingDOMSandrelatedsymptomsas

NSAIDShavethehighestintakeofallmedicationsforeliteandnon-elite

athletes(Tscholl,etal,2016).

2.4NutraceuticalsforthemanagementofDOMS

Overthelast20years,onlyahandfulofnutritionalsupplementshavebeen

investigatedconcerningtheirpotentialinthemanagementandpreventionof

DOMSandrelatedsymptoms.VitaminsCandEarethemostheavilynutrients

investigated(Averyetal.,2003;Bloomer,2004;Childsetal.,2001;McBrideet

al.,1998).Inconsistenciesintheresearchexist,forinstance,bothVitaminEas

wellascertainanti-inflammatorydrugshasshowntoeitheraggravateor

alleviatesymptomsdependingonwhenandhowmuchisused.VitaminE

consumedinhighconcentrationsof1200IU/dpost-exerciseappearsto

aggravateorincreasemuscledamageasdeducedfromobservedincreased

plasmaCKconcentrations,(Averyetal.,2003).Whereassimilardosagesof

vitaminEconsumedforonlyashortaperiodprecedingexerciseinduced

muscledamageappearstohaveaprotectiveeffect(Beaton,Allan,

Tarnopolsky,Tiidus,&Phillips,2002;McBrideetal.,1998).Furthermore,lower

dosesofVitaminE(400IU/d)takenincombinationwithonegramofvitaminC

28

takeneachdayfor14dayspre-andthreedayspostunaccustomedstrenuous

exercisehasshownpotentialtowardsreducingplasmaCKconcentrations

(Bloomer,2004).Moreover,itisunlikelythatthesynergisticeffectofthe

supplementedvitaminCinfluencedtheresults.VitaminCsupplementationof

12.5mg/kg/dlincombinationwithN-AcetylCysteine(NAC)for7dayspost

exercisealsoappearstoaggravateorincreaseplasmaCKconcentrations

(Childsetal.,2001),whereasotherresearchersreportnochangefollowing

vitaminCsupplementationof3g/dpreorpostexercise(Bryer&Goldfarb,

2006;Connolly,Lauzon,Agnew,Dunn,&Reed,2006).Somesupplementshave

shownnoeffectonDOMSandrelatedsymptomssuchasfishoil(Lennetal.,

2002),creatine(Rawsonetal.,2001),chondroitinsulfate(Braunetal.,2005),

bromelain(Stoneetal.,2002),andahighorlowCHOdiet(Ludenetal.,2007;

Whiteetal.,2008).However,somenutrientshaveshownpromisewith

reportedpositiveeffectsatalleviatingonlysome,butnotallsymptomssuchas

ginger(Matsumuraetal.,2015),omega-3fattyacids(BakhtiarTartibian,

Maleki,andAbbasi,2009;BakhtyarTartibian,Maleki,andAbbasi,2011)tart

cherryjuice(Connolly,McHugh,etal.,2006),andpomegranatejuice

(Tromboldetal.,2011).Unfortunately,theFoodandDrugAdministration

(FDA)donotregulatethetherapeuticvalueorqualityofthesesupplements

andthereforeanumberofdietarysupplementslacksufficientscientificdata

oranyatallandarestillavailableforpurchase.Buzzphraseslike,“thesehave

29

yettooffersignificantbenefitsinrigorousstudies,butbecausetheyoffer

benefitsbeyondmusclesoreness”,or“stillprovidingadditional,yet

unexplored,benefitsbeyondantioxidantcapacity,”cansometimesfoulthe

consumerintobeliefofsomebenefitsnotyetsupportedbyrealrigorous

scientificresearch.Alltoooftensupplementsaresoldunderfalsepretenseof

someunknownorpotentialbenefitandthereforeitisadvisablethathealth

careprofessionalsproceedwithextremecautionwhenmaking

recommendationswithoutcriticallyreviewingtherelevantpublishedresearch.

Themostreliablenutraceuticalsarethosewhohavebeentestedbyanumber

ofseparatestudies,conductedatdifferentlabs,withamajorityofsimilar

resultsofsafetyandefficacy.Themajorityofavailablesupplementsmarketed

toimprovehealthand/orexercisetoleranceandperformancearebasedon

theoreticalapplicationsderivedfrombasicorsmallclinicalstudies(Kreideret

al.,2010).AccordingtoKreider,(2010)nutraceuticalsaregenerallyclassified

intooneofthefollowingfourcategories:

1)ApparentlyIneffective–Supplementslackingscientificrationaleorresearch

indicatesittobeineffective.

2)Tooearlytotell–Supplementswithsensibletheory,butlackingsufficient

research.3)PossiblyEffective–Supplementswithinitialsupportingevidence

andtheoreticalrationale,butstillrequiresmorerigorousevaluationonthe

effectoftrainingand/orperformance.

30

4)ApparentlyEffective-Supplementsthatshoweffectiveandsaferesultsfrom

themajorityofresearchstudiesinrelevantpopulations.

Afewproteinsupplementshavebeeninvestigatedthusfarinregards

totheirroleinthemanagementofDOMSandrelatedsymptomsandhave

shownpositiveresultsatimprovedmusclefunctionandsoreness(Hirose,

Sato,Yanagisawa,&Fukubayashi,2013;Nosakaetal.,2006;VanSomerenet

al.,2005).However,again,someresearchersreportonlypositiveeffectsat

alleviatingsome,butnotallassociatedsymptoms(Etheridgeetal.,2008;

Shimomuraetal.,2010).Theinconsistenciesintheresearcharelikelydueto

thetypesofprotein,amountsanddifferentprotocolsofingestion,and

individualdifferencesofbioavailability.Etheridge,(2008)reportedpositive

effectswithregardstorateofforceandpowerdevelopment48hourspost

exerciseafter100gofproteiningestionimmediatelyfollowinga30minute

downhillrunningexerciseprotocol.Thisenhancedrateofrecovery;however,

wasindependentofthecirculatingCKresponseandperceivedmuscle

soreness.By72hours’post-exercisemuscle,functionwasfullyrecovered;

however,perceivedmusclesorenesshadonlyreacheditspeak.Itwouldthen

seemevidentthatevenasingleproteinmealfollowingstrenuousexercise

mightstimulateintramuscularadaptationsthatleadtoimprovementsin

functionalcapacity.Moreover,musclesorenessdoesnotappeartofollowthe

sametemporalsequenceasfunctionalrecovery.Moreover,5gramsofmilk

31

peptidetaken13timeswithinatestingperiod(immediatelybeforethe

exerciseandtwiceadayeachdayfor5consecutivedays)of5dayson

eccentricexercise-inducedmuscledamageshowedthatthemilkpeptidedrink

decreasedpeakCKlevels,MRIvalues,andreportedmusclesoreness(Hiroseet

al.,2013).Itappearsthatproteinconsumedpostexerciseismoreeffective

towardsallocatingDOMSandrelatedsymptoms(Nosakaetal.,2006).

Someaminoacidshaveshownpromiseinreducingthesignsand

symptomsofDOMS.In2005,followingasingleboutofeccentricallybiased

resistanceexerciseinsuccessionwithsupplementationofthreegramsofB-

hydroxy-B-methylbutyrate(HMB)and30milligramsofa-ketoisocaproateeach

dayfor14days,researchersinvestigatedtheselectedsignsandsymptomsof

exercise-inducedmuscledamageincluding:plasmacreatingkinaseactivity,

ROM,%increaseoflimbgirth,1-RMmax,andDOMS.(VanSomerenetal.,

2005).PlasmaCKsignificantlyincreasedfrombaselinepostexerciseinthe

controlgroupandpeaked48hourspostexercise;however,theHMB/KIC

treatmentgroupshowedverylittlechangeinplasmaCKresponse.Although

limbgirthpeaked24hpostexerciseandwasattenuatedintheHMB/KIC

treatmentovertheentire72hpost-exerciseperiod,DOMSwasonly

statisticallylowerintheHMB/KICgroup24hpost-exercise.Therewasa

similartrendintheROMreductionfrompre-exercisebaselinemeasuresover

the72-hourpostexerciseperiod,howevertheHMB/KICgroupwasonly

32

slightlylowerandnosignificantdifferencesinROMdecrementswerenoticed

betweengroups.Interestingly,despitesimilarforcedecrements1-hourpost

exercisetheHMB/KICtreatmentattenuatedthedecrementin1RMoverthe

courseofthe72-hourpost-exerciseperiod.Moreimportantly,althoughnot

statisticallysignificanttherewasaslightincreasein1RMnoted48hourspost

exerciseintheHMB/KICtreatmentgroup.Earlier,Knitter,Panton,

Rathmacher,Petersen,&Sharp,(2000)reportedsimilarfindingsfollowing6

weeksoftrainingand3g/dayofHMB.Theresearchersreportedadecreased

creatinephosphokinaseandLDHfollowingasingle20kmprolongedrunas

comparedtothecontrolgroup.Tajari,Rezaee,&Gheidi,(2010)investigated

theeffectofsupplementationof5gofL-glutaminetaken3timesaweekfor4

weeksonDOMSafter30minergometricexercisebycomparingtwometabolic

enzymes(aldolaseandcreatinekinase)andhipflexorsrangeofmotion.The

studyconsistedof20sedentaryyoungwomen.Aldolasesignificantlyincreased

at36hours’post-exerciseintheexperimentalgroup,butrevertedinthe

control.Creatinekinaseincreasedsignificantlymore36hours’post-exercise

thanintheexperimentalgroup.Inaddition,hipjointROMhadfullyrecovered

by36hourspost-exerciseinthetreatmentgrouponly.ThissuggeststhatL-

glutaminesupplementationmayattenuateDOMSfollowing30minofergo-

metricexerciseforyounguntrainedwomen.Morerecently,Nakhostin-Roohi

andMohammadiAghdam,(2017)assessedtheinfluenceofacuteL-Arginine

33

(3g)supplementationfollowingheavyeccentricexerciseonselectedmarkers

ofDOMS.TheyreportedthatCKandLDHbothsignificantlyincreased,24and

48hours’post-exerciseintheplacebogrouponly.However,althoughboth

groupshadincreasedreportedmusclesoreness,edema,anddecreasedROM,

therewerenosignificantdifferencesbetweengroups.Therefore,

supplementationwithARGafterheavyeccentricexercisemayonlyalleviate

somesymptomsofDOMS.Meroetal.,(2010),reportedthat1.5gofalfa-

hydroxy-isocaproicacid,ametaboliteofthebranched-chainaminoacid

leucine,didshowadecreaseonwholebodyDOMSsymptomsinthe4thweek

oftreatmentascomparedtotheplacebofollowinganormalweeklysoccer

team-trainingschedule.Similarly,supportingMero’sfindings,Katoetal.,

(2015)reportedthatleucine-enrichedessentialaminoacidsupplementation

of1g/kgofbodyweightthirtyminutesbeforeandagain10minutesafter

eccentricexerciseamelioratedmusclesorenessinrats.However,thesolution

administeredcontained40%leucineand60%otheressentialaminoacidsso

theeffectcannotbeentirelyattributedtoleucineingestion.Inaddition,two

weeksoftaurinesupplementation(50mg/kg/day)hasshowntoreducemuscle

soreness,plasmaCKlevels,andincreasestrengthfollowingeccentricexercise

inyoungmen(Silvaetal.,2013).Moreimportantly,taurinesupplementation

didnotaffectpost-exerciseinflammationmarkersnecessaryformuscular

adaptation.Moreover,atherapeuticapplicationoftaurine(0.1g/kg/day)for

34

justthreedayspostexercisehasshowntosignificantlyimprovedpeak

eccentrictorqueofthebiceps(McLeay,Stannard,&Barnes,2017).However,

notreatmenteffectswereobservedaswellasanydifferenceinplasmaCK

levelsbetweentreatments.

Recently,Matsumuraetal.,(2015)showedasignificantreductionin

painfollowinga20-minutesteptestwhen2gramsofgingerwastaken1hour

beforetheexerciseasopposedto1hourbefore.Theresultsalsoshoweda

significantreductionofpain24and48hours’post-exerciseintheplacebo

groupascomparedtoeithertreatmentgroupreceivingginger.Omega-3fatty

acidsupplementationof3000mg/dfor7daysshowednochangeinarm

circumferencefollowinganeccentricarmcurlexercises,butdidreporta

magnitudeofsorenessreductionby15percentintheomega-3trialas

opposedtothecontrol.InthesameyearBakhtyarTartibianetal.,(2011)

publishedsupportingevidencethatOmega3fattyacidsupplementationof

1.8g/dwaseffectiveinamelioratingexercise-inducedinflammatorymarkers

followingtaxingexerciseascomparedtothecontrol.Trombold,(2011),

showedthatpomegranatejuicesupplementation(of250mladayfor8days

pre-exerciseand7dayspostexercise)attenuatedthereductionofisometric

strengthofelbowflexormusclesfollowinganintenseboutofeccentric

exercise,butnotthekneeextensormusclesof17resistancetrainedmen.

Moreover,althoughmusclesorenessratingsweresignificantlyattenuatedin

35

theelbowflexorsduringthe2-168-hourpostexerciseperiodascomparedto

theplacebo,fullrecoveryofreportedsorenesswasnotreportedinthe

pomegranatejuicegroupuntil168hourspostexercise.Theresearchers

concludedthatpolyphenolscontainedwithinthejuicemightberesponsible

fortherecoveryofstrengthinthedaysfollowingeccentricexercise.Three

yearslaterresearchersreportednoextrabenefittoconsuming500mlof

pomegranatejuiceonarmsorlegsfollowingboutsofeccentricexercisethat

resultedinDOMS(Machinetal.,2014).HoweverunlikeTromboldetal.,

(2011),MachinandassociatesdidsuccessfullyobtainexerciseinducedDOMS

inboththekneeextensormusclesandtheelbowflexorsbyimplementing20

minutesofdownhillrunningfollowedby40repetitionsofbilateralisotonic

eccentriccontractionsoftheelbowflexors.Similarly,Ellagitanninfrom

pomegranateextracttaken5dayspreand4dayspost-exerciseresultedin

improvedrecoveryinelbowisometricstrengthfollowingeccentricexercise

(Tromboldetal.,2010).However,nodifferencesinmusclesoreness,plasma

CKlevels,orinflammationmarkerswerenoted.In2006researchersreported

thattartcherryjuicesupplementation(12ouncesoftartcherryjuice

consumptionoftwiceadayfor3dayspre-and4daysposteccentricexercise)

decreasedonlysomeofthesymptomsofexerciseinducedmuscledamage,

(Connolly,McHugh,etal.,2006).Strengthlossandpainratingswere

significantlylessinthecherryjuicetrial;however,relaxedelbowangle(ROM)

36

andreportedmuscletendernesswerenodifferentbetweentrials.This

supplementmostnotablyshowedthatat96hourspostexercisestrengthloss

declined22%intheplacebotrial,butonlyafourpercentdeclinewasobserved

inthecherryjuicetrial.Table2.4.2summarizestheresearchonthedifferent

nutraceuticalsandtheireffectsonDOMSandrelatedsymptoms.

SaffronhasexhibitedsomepotentialtopreventDOMSandallrelated

symptomsandmoreimportantly,researchhasshowntohavesome

performancepreservationeffectfollowingunaccustomedstrenuousexercise

(Meamarbashi&Rajabi,2015).Meamarbashietal,(2015)investigatedthe

preventativeeffectsofsaffronandindomethacinsupplementationon

biochemicalandfunctionalindicatorsofDOMSafter1-sessionof

unaccustomedeccentricexercise.Themostconcerningfindingwasthatthe

saffrontreatmentgroupexhibitedsignificantincreasesinisotonicforceoutput

24,48,and72hoursfollowingexerciseascomparedtobaseline.Mostnotably,

thesaffrontreatmentgroupreacheda63.3%increaseinmaximalisometric

forcecapacityoutputat72hoursfollowingtheexerciseprotocol.Isotonic

forcecapacitysteadilyincreasedfrombaselineat24,48and72hourspost

exercise,butdidnotreachstatisticalsignificance.Furthermore,thesaffron

trialnotonlyexhibitvirtuallynopain,butalsonosignificantdifferenceswere

observedfrombaselinebetweenkneerangeofmotionorthigh

circumference.Lastly,againunlikeboththeplaceboandcontrolgroup,the

37

saffrongroupshowednosignificantincreaseinplasmaCKandLDH

concentrationsfrombaseline.Itisunclearifsaffronisresponsibleforthese

observeddifferencesbecauseafewlimitationsexistintheoriginalwork.

Firstly,thebaselinemaximalisotonicforcemeasuredwasusedtoestablisha

weightloadfortheexerciseprotocol.Thefoursetsof20repetitions

prescribedwerebasedon80%oftheirpreviouslymeasuredmaximumisotonic

forcecapacity.Therewashowevernoformofreliabilitymeasurestakento

provideconfidencethatthismaximalisotonicforcemeasurewasindeed

maximal.Giventhislimitation,theexerciseprotocolprescribedtoinduce

muscledamagemaynothavebeentaxingenough.Inotherwords,ifthis

measurewasnotmaximalatthebeginningofthestudy,thenthefindingsmay

notbereliableastheexerciseprotocolintendedtoinducemuscledamage

wouldhavebeenlessthanadequatetoinduceDOMS.Tocontrolforthis

limitationanEMGcouldbeemployedduringbaselinemeasuresandcouldbe

usedasanindicatortoimprovetheoverallreliabilityofthesemeasures.

Secondly,giventhattherewerethreedifferentgroupsinvolved,aplacebo,an

indomethacin,andasaffrontreatmentgroup,itispossiblethattheresultsare

limitedduetoindividualdifferencesbetweenthegroups.Tocontrolfor

individualdifferencebetweengroupsitmaybebeneficialtohaveonegroup

thatcouldserveastheirowncontrolgroupviaarepeatedmeasuresresearch

designapproach.Thiscouldcontrolforindividualdifferencesbetweenthe

38

groups.Addressingtheselimitationsinthepreviousworkdoneonsaffronand

itspotentialtomanageDOMSwouldseemreasonable.Reproductionor

replicationofthisresearchtailoredaroundaddressingtheselimitationsina

differentpopulationandadifferentlabisvitaltofurtherunderstandthe

potentialforthespicetohelpmanageDOMS.Table2.4.2highlightsthemain

differencesbetweenoriginalpublishedworkonsaffronanditspreventative

effectsonDOMSandthepresentresearch.

39

Table2.4.1Comparisonbetweenthecurrentpublishedworksonsaffronand

thepresentresearch.

OriginalworkbyMeamarbashiand

Rajabi

Presentstudy

Studydesign-A10-day,randomized,double-blind,placebo-controlled,pretest–posttestdesignParticipants–Allparticipantsweremaleuniversitystudentsbetweentheagesof(age:18.2±0.4years).Groups-Therewerethreedifferentgroupsinvolved,aplacebo,indomethacin,andasaffrontreatmentgroup.ExerciseProtocol-Foursetsof20legpressesof80%oftheirpreviouslymeasuredmaximumisotonicforcecapacity.Measures–Voluntarymaxisometricandisotonicmuscularforcevialegpress,bloodsamples,mid-thighcircumference,kneejointROM,andperceivedmusclesoreness.

Studydesign-Adoubleblind,pseudo-random,placebocontrolled;counterbalanceresearchdesignprovidedwithaneight-weekwashoutperiodbetweendatacollectionintervalsParticipants–5femaleand10maleparticipantsbetweentheagesof25.9±3.7and24.4±2.8yearsGroups–Thiswasarepeatedmeasuresdesignsothetwogroupswillserveastheirowncontrolgroup.ExerciseProtocol-Sixsetsof10maximaleccentricvoluntarycontractionsinvolvingmaximalresistanceofthecybexmotorarmataspeedof60degreespersecondfromfullextensiontoflexion.Measures–Maximalvoluntaryisokinetictorque,Maximalvoluntaryisometrictorque,kneeROM,andperceivedmusclesoreness.

40

Table2.4.2summarizestheresearchonthedifferentnutraceuticalsreviewed

andtheireffectsondelayedonsetmusclesorenessandrelatedmeasures.

Study Subjects Supplement Measures MainFindings

Avery

etal.

(2003)

.

18men

(22.7±

4.1

years).

VitaminE

supplementatio

n(1200IU/day)

for21dayspre

exercise.

Body

composition,

DOMS,

Performance,

plasma

Creatine

Kinase(CK)

and

Malondialdeh

yde

Responses.

Nosignificant

diffbetween

placeboand

treatment.

Bailey

etal.

(2004)

.

20men

between

19and

29

years.

Protease

enzyme(325

mg)pftaken

twiceadayfor

1-daypreand3

DOMS,

pressurepain

threshold,and

performance.

The

experimental

group

demonstrated

superior

41

dayspost

exercise.

recoveryof

contractile

functionand

diminished

effectsof

DOMSafter

downhill

running.

Beato

netal.

(2002)

18men

(20.3±

1.7

years).

VitaminE

supplementatio

n(1200IU·d−1)

preexercise.

Performance

andplasma

CK.

Nosignificant

difference

between

placeboand

treatment.

Beck

etal.

(2007)

20men

(21.0+/-

3.1

years).

Protease

enzyme(342mg)

taken1-daypre

and4dayspost

exercise.

Performance,

ROM,arm

circumference

,DOMS,

plasmaCK

activity,and

Myoglobin

(Mb).

Isometric

forearm

flexion

strengthwas

greaterforthe

treatment

groupthanfor

42

theplacebo

group.

Bloom

eret

al.

(2005)

20

resistan

ce

trained

men

(25.1+/-

1.6

years).

Astaxanthin

(4mg)taken21

dayspreand3

dayspost

exercise.

DOMS,muscle

performance,

andplasma

CK.

Nosignificant

diffbetween

placeboand

treatment.

Braun

etal.

(2005)

16

untraine

dmen

between

19-34

years.

Chondroitin

sulfate(600mg)

taken14days

preand2days

postexercise.

DOMS,

performance,

ROM,plasma

CK,

complement

system

fragment.

Inflammatory

markers.

Nosignificant

diffbetween

placeboand

treatment.

43

Bryer

and

Goldfa

rb.

(2006)

18

young

untraine

dmen.

VitaminC(3g/d)

taken14dpre

and4dpost

exercise.

DOMS,elbow

ROM,

performance,

plasmaCK,

totaland

oxidized

glutathione.

Nosignificant

diffbetween

placeboand

treatment.

Childs

etal.

(2001)

14

untraine

dyoung

men

(24.4±

3.6

years).

VitaminC(12.5

mg/kgbody

weight)andNAC

(10mg/kgbody

weight)taken

immediately

afterandeach

dayfor7days

postexercise.

Bleomycin-

Detectable

Iron(BDI),

plasmaCK,

Lactate

Dehydrogenas

e(LDH),Mb,

Superoxide

Dismutase

(SOD),

Selenium-

Dependent

Glutathione

Peroxidase

LDHandCK

activitieswere

elevated

largelyinthe

vitaminCand

NACgroup.

Moreover,the

treatment

grouphad

higherlevelsof

lipidhydro

peroxidesand

8-Iso-PGF2α2

44

(GPX),lipid

hydro

peroxides.

dayspost

exercise.

Conno

llyet

al.

(2006)

24men

and

women.

1000mgof

ascorbicacid3

timesperday

for3dayspre

and5dayspost

exercise.

Performance,

ROM,DOMS,

andpoint

tenderness.

Nosignificant

diffbetween

placeboand

treatment.

Conno

lly,

McHu

gh,

and

Padilla

-

Zakour

.

(2006)

16men

(22+/-4

years).

12fluidounces

ofacherryjuice

blendtwicea

dayfor4days

preand4days’

postexercise.

Performance,

DOMS,muscle

tenderness,

andROM.

Strengthloss

andpainwere

significantly

lessinthe

cherryjuice

trialversus

placebo.

45

Etheri

dge,

Philp,

and

Watt.

(2008)

9men

(21±

1year).

Immediate

ingestionof

100gofprotein

postexercise.

Performance,

DOMS,and

Protein

Carbonyl(PC)

content.

Significant

increaseofthe

recoveryrate

ofisometric

forceand

dynamic

power

production.

Hirose

etal.

(2013)

6

untraine

dmen

(19-22

years).

5gofmilkpep-

tidetaken1h

beforeand

immediately

postexercise

andtwiceaday

for5days.

DOMS,CK,

andMagnetic

Resonance

Imaging(MRI)

T2value.

VASscores,

peakCK,and

T2valuesin

thepeptide

trialwere

significantly

lowerthanthe

controltrial.

Jouris,

McDa

niel,

and

3men

and8

women

(35+/-

Omega-3

supplementatio

nof2,000mg

EPAand1,000

mgDHAfor7

Performance,

DOMS,

swelling,

temperature.

Omega-3

supplementati

onattenuated

DOMS.

46

Weiss.

(2011)

10

years).

dayspreand2

dayspost

exercise.

Kato

etal.

(2015)

Male

rats

Leucine-

enriched

essentialamino

acids(1g/kg

bodyweight)

taken30min

beforeand10

postexercise.

DOMSand

Fractional

SynthesisRate

(FSR).

AminoL40

administration

significantly

mitigatedthe

EC-induced

impairmentof

theFSRand

reducedthe

pawwith

drawl

threshold.

Knitter

etal.

(2000)

8male

and

female

long

distance

runners,

20–50

3gofB-hydroxy-

B-

methylbutyrate

adayfor42days

preexercise.

Maximal

Oxygen

Consumption

(V̇O2max),

body

composition,

Theplacebo-

supplemented

grouphad

significantly

higherLDH

activitylevels

andCPK

47

yearsof

age.

andplasma

CK,LDH.

responsethan

theHMB-

supplemented

group.

Lenn

etal.

(2002)

13men

(22.7+/-

3.92

years)

and9

women

(24.5+/-

5.47

years).

1.8gofomega-3

fattyacidor

120mgof

isoflavones

taken30preand

7dayspost

exercise.

Performance,

DOMS,arm

circumference

,ROM,plasma

CK,

inflammatory

markers,lipid

peroxidation,

andserum

iron.

Nosignificant

difference

between

placeboor

treatment.

Luden,

Saund

ers,

and

Todd.

(2007)

11male

and12

female

cross-

country

runners.

10mL/kgbody

weightofCHO

orCHO+P+A

(0.365g/kgbody

weightofwhey

proteinand

DOMS,plasma

CK,

Performance

measures.

Post

intervention

CKandDOMS

were

significantly

lowerafter

CHO+P+A

48

vitaminsCand

E).

intervention

thanafter

CHO.

Machi

netal.

(2014)

45

young

men

(22.3±

4.1

years).

250mlor500ml

pomegranate

juicetaken4

dayspreand4

days’post

exercise.

Performance,

DOMS,and

Mb.

Both1xand2x

PJCtreatments

resultedin

significantly

higher

isometricknee

extensorand

elbow

extensor

strengththan

PLA.

Matsu

mura,

Zovors

ky,

and

Smolig

20,non-

weight

trained

menand

women.

4gofginger

onceadayfor5-

daypreexercise.

Performance,

circumference

,skin

temperature,

DOMS,plasma

CK,andLDH.

1RM

improved

significantly

48-hourpost

exerciseinthe

ginger

treatment

49

a.

(2015)

groupand

improvedat72

and96honly

intheplacebo

group.Blood

CKcontinued

toincrease

onlyinthe

gingergroup

72and96h

post-exercise.

McLea

v,

Stanna

r,and

Barnes

.

(2017)

10men

(26.5±

6.5

years).

0.1g·kg−1body

weight·day−1of

taurine.

Performance

measures,and

plasmaCK.

Significant

treatment

effects

observedonly

forpeak

eccentric

torque.

Meam

arbash

iand

39

young

men

300mgof

saffronor75mg

ofindomethacin

Performance,

DOMS,knee

ROM,thigh

Thesaffron

intervention

showed

50

Rajabi.

(2015)

(18.2.0

+/-0.4

years).

takeneachday

for7dayspre

and3days’post

exercise.

circumference

,plasmaCK,

andLDH.

significantly

lessCKand

LDH

concentrations

andnodecline

inmaximum

isometricand

isotonicforces

aftereccentric

exercise,buta

significant

declineinthe

isometricforce

inthecontrol

group.There

wasnopain

wasreported

inthesaffron

group,

whereasthe

controldidnot

51

fullyrecover

by72hours

postexercise.

Thigh

circumference

significantly

increasedin

thecontrolat

all-timepoints

anddidnot

changeinthe

saffrongroup.

Controlgroup

showed

significant

decreasein

kneeROMat

all-timepoints

frombaseline

anddidnot

52

changeinthe

saffrongroup.

Mero

etal.

(2010)

15male

soccer

players

(22.1+/-

3.9

years).

500mgofalpha-

hydroxy-

isocaprocicacid

mixedwith

liquidthree

timesadayfor4

weeks.

Body

composition,

DOMS,and

performance.

HICA

significantly

increasedtotal

leanbody

massand

milderDOMS

comparedto

thecontrol.

Nakho

otin-

Roohi,

and

Moha

mmadi

Aghda

m.

(2017)

12

young

women.

3gL-Arginine

oral

supplementatio

nimmediately

postexercise.

DOMS,ROM,

Swelling,

plasmaCK,

LDH,andTotal

Antioxidant

Capacity

(TAC).

Total

antioxidant

capacity

significantly

increased48h

afterexercise

comparedwith

thepre-

exercisejustin

ARGgroup.CK

andLDH

53

significantly

enhanced24

and48hafter

exerciseonly

inthecontrol

group.

Nosak

a,

Sacco,

and

Mawa

tari.

(2006)

38men

between

18and

31years

ofage.

InExperiment1,

7.2gofamino

acidsper

ingested30min

preand

immediately

afterexercise.

Experiment2,

supplements

werealso

ingestedon

additionaleight

occasionsover4

daysafter

exercise.

Performance,

ROM,DOMS,

upperarm

circumference

,plasmaCK,

Aldolase

(ALD),and

Mb.

PlasmaCK,

aldolase,Mb,

andDOMS

were

significantly

lowerforthe

aminoacid

experiment

groupthanthe

placeboin

Experiment2.

54

Rawso

n,

Gunn,

and

Clarks

on.

(2001)

23non–

weight-

trained

men.

5gofcreatine4

timesperday

preexercise.

Performance,

ROM,forearm

circumference

,DOMS,and

bodyMass

(BM).

Nosignificant

diffbetween

placeboand

treatment.

Shimo

mura

etal.

(2010)

12

women

(22.2±

1.6

years).

5.5gofBranch-

ChainAmino

acidstakenpre

andeachdayfor

5dayspost

exercise.

DOMS,

performance,

plasmaCK,

glucose,Free

fattyacids

(FFA's),

lactate,

ammonia,

insulin,

elastase.

DOMSwas

significantly

lowerinthe

treatmenttrial

thaninthe

placebo.

Branched

chainedamino

acid(BCAA)

supplementati

onsuppressed

themuscle-

forcedecrease

to~80%ofthe

55

valuerecorded

underthe

control

conditions.

SerumMb

concentration

increasedin

theplacebo

butnotinthe

BCAAtrial.The

concentration

ofplasma

elastaselevel

was

significantly

higheronlyin

theplacebo

trial.

Silva

etal.

(2013)

21men

(21±6

years).

Taurine50

mg.kgofbody

masstakeneach

DOMS,elbow

ROM,

performance,

Taurine

supplementati

onwasshown

56

dayforfor14

dayspreand7

dayspost

exercise.

perceived

effort,

oxidative

stress,

inflammatory

markers,and

plasmaCKand

LDH.

tosignificantly

increase

concentricand

isometric

strength,

reducemuscle

damage,

DOMS,and

plasmaCKand

oxidative

stress,butit

alteredneither

antioxidant

enzymesnor

the

inflammatory

response.

Stone

etal.

(2002)

20men

and

women

Bromelain300

mgtakenevery

day3timesa

DOMS,ROM,

and

performance.

Nosignificant

diffbetween

placeboor

57

(23±3.2

years).

daypost

exercise.

controland

treatments

Tajari,

Rezae

e,and

Gheidi

.

(2010)

20non-

athletic

women

(22.8±2.

6years).

10gof

Glutaminetaken

3timesaweek

for4weekspre

exercise.

HipROM,

plasmaCK,

andALD.

Significant

higheraldolase

levelincontrol

comparedto

thetreatment

group.

Tartibi

an,

Maleki

,and

Abbasi

.

(2009)

37men

(33.4±

4.2

years).

324mgEPAand

216mgDHAn-3

fattyacidsper

dayover30days

beforeand

during48hours

afterstep

training.

DOMS,thigh

circumference

,kneeROM.

Nosignificant

diffbetween

placeboor

controland

treatment.

Tartibi

an,

Maleki

,and

Abbasi

45

untraine

dmen

(29.7±

324mgEPAand

216mgDHAn-3

fattyacidsper

dayover30days

beforeand

PlasmaCK,

LDH,and

inflammation

markers.

Significantly

lessIL-6,CK,

andMbforthe

experimental

groupat24

58

.

(2011)

6.6

years).

during48hours

afterstep

training.

and48hours

afterexercise

thanthe

control.

Tromb

old,

Barnes

,

Chitchl

ey,

and

Coyle.

(2010)

16

recreati

onally

active

men

(24.2+/-

1.4

years)

500mlof

pomegranate

extracttaken

twiceadayfor5

dayspreand4

dayspost

exercise.

Performance,

DOMS.

Recoveryof

strength

duringthe24-

to48-hperiod

wasmore

rapidinthe

treatment

group.

Tromb

old,

Reinfel

d,

Casler,

and

Coyle.

(2011)

17

resistan

ce

trained

men

(21.9±

2.4

years).

500mlof

pomegranate

juiceingested8

dayspreand

immediately

postexercise.

Performance,

DOMS.

Onlyisometric

elbowflexion

strengthwas

significantly

greaterwith

thetreatment

comparedwith

thatwiththe

59

control.Elbow

flexorsoreness

wasreducedin

treatment

groupat48

and72hours

postexercise.

Van

Somer

en,

Edwar

ds,

and

Howat

son.

(2005)

Eight

men

(23.0+/-

4years).

3gofB-hydroxy-

B-

methylbutyrate

adayfor14days

preexercise.

1-RM,plasma

CK,DOMS,

Limbgirth,

ROM.

HMB/KIC

supplementati

onattenuated

theCK

response,the

percentage

decrementin

1RM,andthe

percentage

increasein

limbgirth.

Moreover,

DOMSwas

lowerat24h

60

post-exercise

forthe

treatment

group.

Volek

etal.

(2002)

10

weight-

trained

men

(23.7+/-

2.3

years).

2gof

carnitine/day

takenfor21

dayspreand6

days’post

exercise.

DOMS,MRI,

Mb,lactate,

purine

catabolism,

freeradical

generation,

andcytosolic

proteins.

Plasma

markersof

purine

catabolismand

circulating

cytosolic

proteinswere

significantly

attenuatedby

carnitine

supplementati

on.

White

etal.

(2008)

27untrai

nedmen

(21±3

years).

CHO/protein

drinktaken

immediately

beforeorafter

exercise.

DOMS,

performance,

andplasma

CK.

Timingof

CHO/Protein

ingestionhad

noeffect.

61

2.5Whyisitimportant?

Thisresearchisvaluableinformationforthoseconcernedabout

musculardiscomfortandpainthatcanbeassociatedwithsports,training,and

strenuousactivity.Currently,(NSAIDS)suchasibuprofenandindomethacin

arecommonlyusedtohelpmanagemusculardiscomfortandrestorationof

physicalfunction.TherelianceofthesedrugsforthemanagementofDOMS

canleadtooveruseandincreasedriskofpotentialsideeffectssuchas

stomachulcers,hepatic,andrenaltoxicity(Tscholletal.,2016).Thisisa

growingconcernforthosemanagingDOMSandrelatedsymptomsasNSAIDS

havethehighestintakeofallmedicationsforeliteandnon-eliteathletes

(Tscholl,etal,2016).Moreover,emergingevidenceintheliteraturesuggests

thatblockingtheinflammationprocessduringtrainingmaylimitmuscle

adaptationsforathletesorthosewelltrained(Schoenfeld,2012).Howevera

differentstoryemergesfortheelderlyasmanaginginflammationwith

standardoverthecounterdosesofNSAIDSappearstohavesomebenefit

towardsimprovedperformance,managingmusclesoreness,andeven

improvedmuscleadaptation(Trappeetal.,2010,2002).Ifsaffron

demonstratestobeaseffectiveasoverthecounterNSAIDS,atmanaging

musclesorenessandpreservingperformance,furtherresearchwouldbe

calledfortoidentifyanylimitationsofmuscleadaptivecapacityforthose

62

interestedinlong-termuse.Ifperformance-limitingtrainingeffectsofsaffron

areidentifiedthantheapplicationofthissupplementwouldbebestsuitedfor

competitionswherepreservationofperformancewouldoutweighthe

attainmentofperformancegains.Moreover,providedthatsaffronshowsan

effecttowardsthepreventionofDOMSandrelatedperformancereductions,it

maybemoresuitablethenNSAIDSforsedentaryindividualsand/ortheelderly

concernedwithmusclediscomfortenablingasmoothertransitiontoamore

activelifestyle.

2.6Saffron;whatisitandwhatisinit?

Saffronisawidelyusedcookingspicethatisharvestedfromthecrocus

sativusflowerandhasbeentraditionallyusedasamedicineformanydisease

conditions.Saffroncontainspotentantioxidantcompounds(crocinand

crocetin)thathavebeenrecentlyshowntohaveanti-inflammatoryandanti-

nociceptiveproperties(Hosseinzadeh&Younesi,2002).Saffroncontainsa

varietyofcompoundswhileitscompositionisdependentontheplantgrowth

andsoilconditions.Compositionanalysesofsaffronhaveidentifiedthat

saffronisapproximately12-15%CHO,8-15%moisture,10-14%protein,5-7%

mineral,5-7%,oils5-9%,volatileoils0.3-0.8,and4%fibreandvitamins

riboflavinandthiamine(Kumaretal.,2008;Rios,Recio,Giner,&Manez,

1996).Inadditiontonumerouspotentialhealthpromotingcompoundsin

saffroncrocin,crocetin,picrocrocin,andsafranalarethemain4bioactive

63

compoundsandareresponsibleforthecolor,color,taste,andaromaofthe

spice.

Thereislittle,butsomesupportingevidenceintheliteratureonsaffron

anditspotentialtopreventmuscledamageandpromoterecovery.Moreover,

crocetinhasbeenshowntoincreasetheoverallrelativegrowthrateofnormal

ratmuscle-derivedcells(Wilkinsetal.,1977).Similarly,highdosesofsaffron,

100mg/kgadministeredfor5consecutivedaystorats,resultedinasignificant

increaseinbodyweightdespiteareduceddailydiet(Asdaq&Inamdar,2010).

Thisincreasedgrowthratemaybeattributedtocrocetinanditsabilityto

stronglybindtoalbumin,whichappearstopromoteoxygentransportinto

tissues(Miller,Willett,Moss,Miller,&Belinka,1982).Moreover,Crocetinhas

beenshowntoenhancewhole-bodyoxygenconsumptioninratsthatwere

bledout40%oftheirtotalbloodvolume,andconsequentiallyimprovedtheir

overallsurvivalrates(Gainer,Rudolph,&Caraway,1993).Additionally,saffron

containssomeprotein,butsaffronalsoappearstoimprovethedigestionof

proteinsviastimulatingthesecretionofgastricacidandpepsinoutputs

(Nabavizadeh,Salimi,Sadroleslami,Karimian,&Vahedian,2009).Thisaction

ormechanismmayinsomewayberelatedtotheimprovedfunctional

capacityobservedfollowingproteasesupplementationfollowingexercise

inducedmuscledamage(Becketal.,2007;Milleretal.,2004).Inotherwords,

saffron’sabilitytoenhancedigestionofproteinssimilartothatofprotease

64

supplementationmayhaveasimilarprotectiveeffectonmusclefunction

followingexerciseinducedmuscledamage.

Saffronhasshowntobestrongerthanthesumofitspartsasthe

additiveand/orsynergisticeffectsamongthemanyphytochemicalsappearsto

morepowerfulandenhanceitseffect.Manyresearchershavereportedthat

consumptionofwholesaffronissuperiortoitsisolatedandevenhighly

concentratedindividualcomponents.Forexample;wholesaffronhasshown

tobeamorepowerfultreatmentforinsomniaandanxiety(Hosseinzadeh&

Noraei,2009),andcancer(Liu,2004)thananyofitsisolatedandconcentrated

components.Inaddition,arecentstudydemonstratedthatsaffronwas

superiortocrocinwithrespecttoitsantioxidantactivityandprotectiveeffect

ofdiet-inducedhyperlipidemiainrats(Asdaq&Inamdar,2010).More

importantly,amongthetreatedgroups,saffroninitshighestdose(100mg/kg

p.o)wasreportedtobethemosteffectiveinmaintainingcellmembrane

integrity.Similarresultsofsaffronsupplementationanditsprotectiveeffect

onmembraneintegrityhavebeenreportedonratspermmembraneintegrity

(Vaezetal.,2014)andredbloodcellmembraneintegrity(Meamarbashi&

Rajabi,2013).Theprotectiveeffectofsaffrononmembraneintegrityislikely

duetoitsantioxidant/anti-inflammatorycapacityandagainappearstobe

strongerwhentakenwholeratherthaninitsconcentratedisolated

65

components;thisislikelyduetothepossiblesynergisticand/oradditive

effectsofthesecomponents(phytochemicals).

2.6.1TheBioactiveCompoundsinSaffron:Crocin,Crocetin,Picrocrocin,and

Safranal.

1)Crocin–Isahighlywater-solublecarotenoidderivativeorpigment(ester).

Inadditiontobeinganexcellentcoloringagent,Crocinalsoexhibits

antioxidanteffectstowardscellsandtissuesagainstoxidationandstress

(Melnyk,Wang,&Marcone,2010).

2)Crocetin–Highlywater-solublecarotenoids.Crocetinisthemostheavily

studiedactivecompoundisolatedfromsaffron.Thiscompoundhasalso

shownhighantioxidantpotential(Razak,Hamzah,Yee,Kadir,&Nayan,2017).

Crocetinhasalsobeenshowntoincreasetherelativegrowthofnormalrat

muscle-derivedcells(Wilkinsetal.,1977).Similarly,highdosesofsaffron

ingestionof100mg/kgadministeredfor5consecutivedaystoratsresultedin

asignificantincreaseinbodyweightdespiteareduceddailydiet(Asdaq&

Inamdar,2010).Theincreasedrelativegrowthrateassociatedwithcrocetin

ingestionmaybeduetoincreasedoxygentransportintheribosomal-

microsomalfraction.Researchersdemonstratedthatcrocetinbindsstronglyto

albumintothesamebindingsitesthatareemployedbyfreefattyacids(Miller

etal.,1982).Inadditiontoalbuminbeingtheprimaryvehicleforcrocetin,the

researchersalsohypothesizedthatthemechanismbywhichcrocetinincreases

66

oxygendiffusivitymaybedirectconsequenceofcrocetinbindingtoalbumin.

Moreimportantly,Gainer&Chisolm,(1974)showedthatcrocetincouldbring

aboutlargeincreasesinoxygendiffusivityintheplasmadespitethepresence

ofincreasedplasmaproteins.Thisisaninterestingfindingbecauseahigher

levelofplasmaproteinsusuallyresultsinalargereductioninoxygen

diffusivitywithinthebloodplasma(Chisolm,Gainer,Stoner,&Gainer,1972).

Thebindingofcrocetintoalbuminappearstopreventoroffsetthereduced

oxygendiffusivitythatwouldotherwiseoccurwithelevatedplasmaproteins.

Inaddition,supportingresearchdemonstratedthatcrocetinsupplementation

increasedoverallsurvivalratesinratsthatwerebled40%oftheirtotalblood

volume.Theresultsindicatedthatcrocetinwasresponsibleforincreased

whole-bodyoxygenconsumption(includingmuscle)andsurvivalrates(Gainer

etal.,1993).

3)Picrocrocin-Picrocrocinisthesecondmostabundantcompoundinsaffron

(byweight)andisprimarilyresponsibleforthebittertasteofsaffron.

Picrocrocinandcrocinaswellassaffanalareformedfromthecleavageofa

carotenoidzeoxanthin(Razaketal.,2017).

4)Safranalisthemostabundantessentialoilofthe160volatileoil

componentsidentifiedinsaffron,andaccountsforapproximately30-70%of

thetotalessentialoilavailable.Safranalismainlyresponsibleforthearomaor

67

smellofsaffron.Thiscompoundhasalsoshownhighantioxidantpotential

(Melnyketal.,2010).

Researchofthemetabolicpathwayoforallyadministeredcrocetinand

crocinsinmiceindicatethattheyarepartlymetabolizedtoestertype

glucuronideconjugatesinboththeintestinalmucosaandintheliverviathe

portalvein(Nakano,Takahashi,&Nagao,2005)Studiesshowthatcrocinisnot

readilyabsorbedfromthegastrointestinaltractuntilitishydrolyzedto

crocetin.In2004,Jinreportedfollowinganobservedreductionofabsorption

ofcrocin-1invariousintestinesegments,thatcrocincouldn’tbeabsorbed

equallythroughoutthewholeintestinaltract.Moreover,starchmaybeableto

enhanceabsorptionofcrocinbyreactionsbetweensalivarynitriteinthe

gastriclumenresultingintheformationofstarch/crocincomplexes(Hirota,

Takahama,2013).Thereductionofnitritetonitricoxidebycrocinformingthis

startch/crocincomplexwasattributedtoenhanceintestinalabsorption.In

vitrostudiesindicatethatcrocinsgethydrolyzedintheintestineto

deglycosylatedtrans-crocetin,whichispassivelydiffusedwithinashorttime

acrosstheintestinalbarriermakingitswayslowlyacrossthebloodbrain

barriertothecentralnervoussystem(Lautenschlageretal.,2015)intestinal

formation2015).However,thesestudiesarelimitedtoanimalstudiesand

maynotbegeneralizabletothehumanpopulation.

68

2.6.2ToxicityandSideEffectsofSaffron

Reportsofsaffrontoxicologyandsafetyaresomewhatcontroversial.

Althoughdailydosesofupto1.5gofsaffronarethoughttobesafe,someside

effectshavebeendocumentedfrominjectionsof1.2to2gperaveragebody

weightofsaffron(Schmidt,Betti,&Hensel,2007).However,Schmidt,(2007)

explainsthatsomecontraryfindingsreportnoadverseeffectsofingestionof

upto4gofsaffronperdayforseveraldays.Thesedifferentfindingsmaybe

duetothesaffronusedduringtheexperiments.Manyofthesereported

contraryresultsarefromGermanywhereasimilarplantcalledmeadow

saffronisabundant.Itispossiblethatthesestudieswerenotconductedon

thesamesaffron,asitisnotclearlyspecifiedintheliterature.Dosesof

between5and10gareconsidereddangerousandmaycausenausea,

decreasedappetite,vomiting,diarrhea,vertigo,andbleedingofthe

gastrointestinalmucosaanduterus;ingestionofabove10gmayinducelabor

andabortionandaround20gisconsideredlethalSchmidt,(2007).Inaddition,

saffronhasshowntoproduceverylittleallergenicriskandonlyonecaseof

anaphylacticreactiontosaffronhasbeenreported(Lucas,Hallagan,&Taylor,

2001).Overallresearchersconsidersaffrontobesafeforconsumptionandthe

amountsusedindailyfoodconsumptionandthatoftheproposedresearchis

muchlowerthananydosesassociatedwithundesirablesideeffects(Bisset,

1994).

69

2.6.3SaffronQualityandGrading

Theinternationalorganizationofstandardizationisaprivate

(nongovernmental)worldwidecollectivefederationofnationalstandard

bodies(ISO)dedicatedtocreatingandimplementinguniformstandardsfor

internationalexchangeandservices.TheISOrecognizesvaryingqualitiesof

saffronviaspecifictestmethodsandcategorizesthequalityonthedifferent

concentrationsofcrocin,picrocrocinandsafranal(“ISO3632-1:2011,”).This

twopartstandardmethodisusedtoverifynoexternalmatterhasbeenadded

andthattheconcentrationsofthebioactivecompoundspreviouslymentioned

areinappropriateconcentrations.Obviously,theISOregardshigher

concentrationsofthesecompoundsasahigherqualityproductand

classificationissetonaminimumrequirementofeachqualityandisregarded

asISO3632.Thisclassificationsystemwasusedtoestablish4categorieswith

category1representingthehighestqualitysaffron(Rasaneh,2000).

2.7IsokineticDynamometry

Thetermisokineticwasfirstintroducedwhenthecybex1

dynamometerwasfirstdevelopedinthe1960’s,howevermostofthe

literaturesurroundingisokineticdeviceshasfocusedaroundthenewly

developedcybex11.Isokineticdevicesmeasuretorquewhilethelimbis

movingandthereforehavebeenreferredtoas‘dynamictesting’.‘Isokinetics’

isdefinedasadynamicmuscularcontractionwherethevelocityiscontrolled

70

forandmadeconstantbyacontrolmechanism(Trestle,Hislop,Moffboid,

Hofkosh,&Lowmxn,1967).Thetotalresistanceappliedtothedeviceisequal

totheappliedmusculartorqueoverthegivenrangeofmotion.Anyincreasein

musculartorqueisobservedwhenthelimbreachesacertainpre-setvelocity

thatengagesacontrolmechanism,thusanyforceabovethislevelresultsinan

equalmagnituderesistiveforcebythecontrolmechanismofthe

dynamometer(Moffroid,1969).Becauseofdifferentjointanglesand

biomechanicalpropertiesofthemusculoskeletalsystem,peakmuscularforce

mayvary.Therefore,itisimportantthattheaxisofrotationoftheattached

limbbealignedwiththemechanicalaxisofthemachine.Isokineticdevices

notonlyareusedtomeasuretorque,buttheycanalsomeasurethe

accompanyingROMasafunctionoftime(Perrin,1993).Inaddition,work

measurementscanbederivedfromtheangulardisplacementofthetorque

valuesandtheforce,torque,workandpowermeasurementsarethe

parametersthatcanbeobtainedfromisokineticdevices(Haffajee,Moritz,&

Svantesson,1972).

Inshort,musclestrengthcanbeevaluatedviaisokinetic

dynamometersbyusingisometricandisokineticmuscularcontractions.This

allowsthesubjecttogenerateasmuchforceaspossibleataconstantvelocity.

Isometricassessmentisusedtoevaluatethemuscleforceagainstafixed

resistancewithoutmovementofthelimbandisokineticassessmentisusedto

71

evaluatethemuscleforcethroughafixedrangeofmotionataconstant

velocity(Perrin,1993).Inconclusion,itisgenerallyacceptedthattheuseof

isokineticmeasurementstomeasuretorqueisappropriateaslongascareis

takenwithrespecttotheaxisalignment.Moreover,foraccurateassessment

ofmusclefunction,onlyconstantvelocitydatashouldbeanalyzed.

2.7.1GravitationalEffectsonIsokineticMovements

Isokinetictestinginaverticalplaneinvolvesmuscularforcesandthe

forceofgravitythatisgeneratedviatheweightofthelimb.Therefore,the

torquemeasuredisnottheactualtorque,buttheresultanttorquegenerated

bybothforces(Herzog,1988;Winter,Wells,&Orr,1981).NelsonandDuncan,

(1983)presentedasimplifiedmethodforthecomputationofthegravitational

torqueduringkneeextension/flexionmovements.Theweightofthelimb-

leverarmisusedtocorrectthegravitationaltorqueateveryangularposition.

Thegravitationaltorquecorrectionalfactoristhenaddedtothemaximum

torqueproducedbythemusclesapposedfromgravity(quadriceps)and

subtractedfromtherecordedtorqueproducedbymusclegroupsfacilitatedby

gravity(hamstrings).

2.7.2InertialEffectsonIsokineticMovements

Thetorquemeasuredduringisokinetictestingoftencontainsa

prominentinitialspikethatcanbefollowedbyoscillationsofdecreasing

72

amplitude(Sapega,Nicholas,Sokolow,&Saraniti,1982).Thisovershootis

sometimesreferredtoas‘torqueovershoot’andalwaysispresentintheinitial

partofthemovement.Sincetheoscillationsrepresentalternatingperiodsof

accelerationanddecelerations,asimplemethodtoovercomethisovershoot

istoonlyusetorquedatafromconstantvelocityperiodsofthemovement

(Osternig,Sawhill,Bates,&Hamill,1982).This‘artifact’freedatacanbe

obtainedfromanalysisofthemovementwheretheangularvelocityremains

constantandequalinmagnitudewiththepresentvelocitysettingofthe

dynamometer.However,iftheovershootisnoteliminatedoraccountedforas

anartifact,itcanbemisinterpretedasapeaktorquemeasure.Formeasures

ofvaryingvelocity,adampsettinghasbeenemployedforthecybex11and

usedtodecreasethe‘overshoot’(Sinacore,Rothstein,Delitto,&Rose,1983).

Thedampappearstobeaconvenientmethodtoguardagainstovershooting,

butitdoescausesignificantchangesinthetorquecurvesoutput.Thetorque

valuesarereducedorbecomesmallerasthedampsettingisincreasedand

shouldbeconsideredbeforeuse.

2.7.3IsokineticandIsometricMaximumTorqueMeasures

Ahandfuloftestingprotocolshavebeenusedfortheassessmentof

maximaltorquemeasures.Themaindifferencebetweenprotocolsisthetotal

numberofrepetitionsneededtoachievemaximaltorqueoutput.For

instance;Sawhill,Bates,Osternig,&Hamill,(1982)suggestedthat4maximal

73

repetitionsarerequiredtoobtainreliablemaximaltorquemeasuresduring

isokinetictestingvelocitiesrangingfrom200-400degreespersecond.

However,Patton&Duggan,(1985)havedefinedthemaximumtorqueasthe

meantorquetakenfrom5maximalrepetitionsand/ortheaverageof3

maximumrepetitions.Formaximalisometriccontractionswiththeknee

extensors,thehighesttorquemeasuresappeartocoincideat50-70degreesof

flexion(Haffajee,Moritz,&Svantesson,1972).

Maximalvoluntarycontractions(MVC’s)oftorquemeasuresprovide

thebestmethodsforquantifyingmuscleinjuryforhumansubjectsaccording

toareviewarticleontoolsusedinthestudyofeccentriccontractionin

inducedinjury(Warren,Lowe,&Armstrong,1999).ThereliabilityofMVC

torquemeasuresisgenerallyhighwithreportedintra-classcorrelation

coefficientsof≥85(Sale,1987;Tesch,Dudley,Duvoisin,Hather,&Harris,

1990).However,effortstoevaluatethereliabilityofsuchMVCbyEMGand

electricalstimulationwereonlyemployedby7ofthe58articlesreviewed.The

authorsconcludedforfutureresearchthatmoreeffortshouldbemadeto

includeothercontractileparametersthatmaymoredirectlyreflectinjury-

inducedfunctionalparameters.Theyalsoconcludedthatmeasurementsof

MVCtorquetakenunderisometricandisokineticconditionsandROMwould

bethemostvaluableandreliablemeansofquantifyingthefunctional

decrementsresultingfromexercise-inducedmuscleinjury.Othercommonly

74

usedmarkersusedtoquantifymuscleinjuryincludereleaseofmyofibre

proteins(CK,LDH),soreness,andhistopathology.Becausethesemarkersdo

notfollowthesametime-courseoffunctionlossandsorenessandagain

correlatepoorlywithchangesinmusclefunction,thesemarkersmaynotbe

thebestmethodstoquantifythedegreeofmuscledamage(Warrenetal.,

1999)

Thisresearchaimedtohelpunderstandthepreventativeeffectsof10-

daysupplementationwith300mgofsaffrononexerciseinducedDOMSby

evaluationofchangesofmaximalisometricandisokinetictorquemeasures,

rangeofmotion,andreportedmusclesoreness.Ifthequalitiesofsaffron

permitcellularprotectionofsomemannerand/oraspeededrecoveryfrom

trauma,itcouldserveasaninvaluablesupplementforthemanagementof

DOMSandrelatedsymptoms.

75

3.0 Methods

AlldatacollectiontookplaceintheAndrewandMarjorieMcCain

HumanPerformanceLaboratorylocatedattheRichardJ.CURRIECenteratthe

UniversityOfNewBrunswick(UNB).

3.1Participants

ParticipantswererecruitedviaadvertisementontheUNBcampusand

webpagefromthestudentundergraduatepopulationatUNBFredericton.A

priorpoweranalysisindicatedthatforarepeatedmeasuresresearchdesign,a

sampleof16maleand16femalesubjectswouldbeappropriateforsufficient

statisticalpower.However,atotalof13healthymalesand5females

volunteeredforthestudywithameanageof25.9±3.7and24.4±2.8years.

Oftheserecruits,threemales’datawereomittedfromtheisokinetictorque

analysisandonefromboththeROMandreportedmusclepainmeasures.The

firstwasomittedfromallthreedependentvariablesbecausetheparticipant

wasunabletomakeittothelabtocompletetheir48-hourmeasurements.

Thesecondwasomittedduetoasoftwaremalfunction.Inaddition,thethird

wasomittedbecausethecycleergometerusedwassuspectednottobe

adequateandmayhaveresultedinabaselinemeasurelessthantheactual

baseline.Theseatofthebikewasunstable,andwouldtipeitherforwardor

backwarddependingonhowtheparticipantdistributedtheirweight.Itwas

76

verylikelythattheparticipantusedhisweighttostabilizehimselfwiththe

bikepetalsandthisresultedinalessthanoptimalwarm-up.Theexaminer

quicklycorrectedthisbyacquiringpermissiontouseacycleergometerlocated

attheCurryCentreStrengthCentreoneflooraboveAndrewandMarjorie

McCainHumanPerformanceLaboratory.Table3.1illustratestheselected

doubleblind,pseudo-random,placebocontrolled;counterbalanceresearch

designprovidedwithasixtoeight-weekwashoutperiodbetweendata

collectionintervals.

Table3.1Pseudo-RandomCounterbalanceResearchDesign

Totalingroup=16FirstDatacollection

Datacollectionfollowinga6-8weekwashoutperiod

Subgroup1n=8 TreatmentA TreatmentBPosttestSubgroup2n=8 TreatmentB TreatmentAPosttest

Exclusioncriteriawereestablishedfromself-reportedresultsfroma

PhysicalActivityReadinessQuestionnaire(PAR-Q)formandPhysicalActivity

andSedentaryBehaviorQuestionnaire.Thosewhoadmittedtoparticipationin

anylowerbodyresistancetrainingwithinthelast3months.Lastly,anypastor

presentinjurythatwouldremotelycauseanyrisktotheparticipantwascause

forexclusionsuchasanyanteriororposteriorcruciateligamentinjury,

meniscaltears,tendontears,tendonitis,painetc.Participantswererequested

torefrainfromanynutritionalsupplementsincludingvitaminA,C,andany

non-prescriptionmedicinesforoneweekbeforeandduringthestudyanda

77

dietarylogsheetwasprovided.Caffeineintakewasaconcernbecauseithas

beenshowntoincreasecalciumreleasefromtheSRandmayenhance

muscularfunctionfollowingunaccustomedstrenuousexerciseand

consequentiallyimprovementofpreservedperformance(Hurley,Hatfield,&

Riebe,2013).However,asrecommendedbytheAmericanCollegeofSports

Medicine(ACSM)guidelinesforexercisetestingandprescription,participants

wereaskedtonotconsumecaffeine2hoursbeforemaximalforcemeasures

ondays6-10ofsupplementation(Thompson,Arena,Riebe,&Pescatello,

2013).Participantsweregivenadequateinformationregardingallprocedures,

risks,andmeritsoftheresearchwereaskedtoreadandsignaninformed

consentletterbeforehand(AppendixA.TheUniversityResearchEthicsBoard

approvedthisresearch(REB2016-127).

3.2SaffronManufacturing

ThesaffronselectedwasshippeddirectlytotheKeswickCompounding

GuardianPharmacyinKeswick,N.B.formanufacturing.Itwasthenprocessed

intonon-chewabletabletsthatwereindistinguishablefromtheplacebounless

chewed.ThetreatmentandplaceboweretobelabeledAorBandtheidentity

ofthesaffronortreatmentwerenotdisclosedtotheresearchersinvolved

untilafterdatacollection.Theplaceboandsaffronpillswerepreparedinsuch

afashionthattheywereindistinguishableunlesschewed.Theplacebo

containedmicrocrystallinecellulosethatwasdyedredtomimicthecolorof

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thesaffronandincasedinplantsourcedgelatincapsules.Boththeplacebo

andsaffronpillswerescentedwithcardamomandcinnamonessentialoils.

Theparticipantswereinstructedtoswallowthepillswithflavoredjuice.

3.3Instrumentation

ACybexHUMACNorm(CSMI,USAInc.)isokineticdynamometerwas

employedtoassessmaximaltorquemeasuresandfortheinducementof

DOMS.Givenafixedvelocityandjointangle,thisapparatusisgenerallyused

toquantifymuscularworkandpowercapacityandmostoftenisusedto

measuretorqueproduction.Thecybex11isokineticdynamometerhasbeen

showncontrolvelocitywellwithin2%,atlowervelocities,30and60deg/sec

andwithin1%accuracyofrecordedpeaktorquewiththedampingsetatthe

highestlevel(Bemben,Grump,&Massey,1988).

Figure3.1-Testingprotocol.ACybexHUMACNorm(CSMI,USAInc.)

isokineticdynamometerusedtostimulateexerciseinducedmuscledamage

andforpre-andpost-maximalisometricandisokineticforcemeasures.

79

3.4Procedures

Allsubjectswereprovidedwith1familiarizationsessionofthelaband

theexperimentalprocedures.Datacollectionbeganaftersubjectssignedthe

informedconsentformaswellasthephysicalactivityquestionnaireandParQ

plusform.ThedominantlegwasidentifiedviatheWaterlooFootedness

Questionnaire,whichhasbeenvalidatedinpreviousresearch(Kang&Harris,

2000).Thekneewaseccentricallyflexedwhilethepatientwasinstructedto

extendtheirkneemaximallyagainsttheresistanceofthecybexmotorarm

untiltheirkneereachedfullflexion.Allmeasuresweretakenbeforeand24,

48,and72hoursfollowingtheexerciseinducingmuscledamageprotocol.

IndependentanddependentvariablesarelistedinTable3.2.Anoverviewof

theexperimentalprotocolshowingthefamiliarizationsessionsandexercise

protocolsisprovidedinTable3.3.Table3.3illustratestheoutlineofthe

researchdesigninvolving10daysofsupplementation,theexerciseprotocol,

andselectedindirectmeasuresofDOMS.

Table3.2Theindependentanddependentvariablesofinterest

Independentvariables DependentVariablesTreatment(Saffron)Time(Pre/Post)

IsometricTorque(N)IsokineticTorque(N)ROMofkneeMuscleSoreness

80

Table3.3Overviewofexperimentalprotocol

-Beforedata

collectionbegan,one-

familiarizationsessionofprotocolexpectationsand

equipmentsetup.

Day1-6

Baselinemeasuresofthefollowingtaken:

ROM

MuscleSoreness

MaxIsometricandIsokineticpeaktorque

*ExerciseProtocolExecution

Postexercise,peaktorquemeasures

Day7

*24hourspostexercise

Measuresof:

ROM

MuscleSoreness

Max

IsometricandIsokinetic

peaktorque

Day8

*48hourspostexercise

Measuresof:

ROM

MuscleSoreness

Max

IsometricandIsokinetic

peaktorque

Day9

*72hourspostexercise

Measuresof:

ROM

MuscleSoreness

Max

Isometricand

Isokineticpeaktorque

Day10

3.5.1MaximalIsometricandIsokineticTorqueAssessment

Thecybexisokineticdynamometerwasusedtoquantifyconcentricand

eccentricmaximalisokineticpeaktorquemeasuresofthekneeextensorsat

baselineandagain24,48,and72hourspostexercise.Subjectswereseated

andstrappedinwitharmsfoldedinfrontoftheircheststolimitextraneous

81

movements.Theparticipant’shipanglewassetat90degreeswhileseated.

Thekneejointcenterwasalignedwiththecenterofthedynamometer’s

powershaftwiththeanatomicalzerosetatakneeangleofzeroorfull

extension.Themassofthelegwasweighedbythedynamometerandthen

usedtocorrectforgravitationaleffects.Maximalisokineticforcecapacitywas

measuredatavelocityof60degreesperwithinarangeof0-120degrees,

where0degreesreferstofullextensionofthekneejoint.Threesuccessive

maximalcontractionswereperformedwith3minutesofrestprovided

betweenevents.Thehighestpeaktorqueofthethreemaximaltorque

measureswasusedforanalysis.Secondly,maximalisometricpeakforcewas

performedat50degreesbelowkneeextension.Again,3minutesofrest

betweentrialswereprovidedandthehighestpeaktorqueofthethree

maximalcontractionswasusedforanalysis.

Duetoaresearchprotocoloversight,maximalisometrictorque

measureswerenottakenatthesamejointangle.Itwasnoticedbythe

examinerpartwaythroughthestudyaftercompleting9malesand2female

peakisometrictorquemeasuresthatthejointangleofthemoreflexible

participantswaspositionedgreaterthan50degreesonthecybex.Thejoint

anglewasapproximately5to10degreesoffdependingonthedegreeof

flexibilityaboveandbeyondorbelowtheangleoftheparticipant’sknee

extension.Thecybexestablishesazerodegreestartingpointbasedonthefull

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extensionoftheleg.Ifdifferinganglesoftheparticipant’sfullkneeextension

wereusedtoestablishazero,itwouldthenselect50degreesfromthezero

degreestartangle.Thiswouldcausethecybextoselecta50-degreejoint

anglefromtheestablishedzeroofthefullextension.Thisequipmentprotocol

oversightmayhaveaffectedthebaseline,24,48,or72-hourisometrictorque

measuresof7maleand2femaleparticipants.Thisoversightwascorrectedby

theexaminerpartwaythroughthestudybyrestrictingthelegextensionROM

tothesamerangeinordertohavetheangleselectedthesameandtherefore

comparable.Althoughthisoversightlimitsourabilitytocompareisometric

torquemeasuresconfidently,webelievethattheeffectoftheoveralltorque

producedbetweenmeasureswouldbeminimalduetothecounterbalancing

designofthestudy.ThemaximalisometricmeasuresarelistedinAppendixD.

3.5.2ROMAssessment

Therangeofmotionassessmentwasperformed,viaagoniometer

guidedbytheuseofsemi-permanentmarkeduniversallandmarks,whilethe

subjectlaysupineonaplinthwiththeirkneefullyextended.Fromthefull

extensionposition,apassiveflexionwasperformedatveryslowangular

velocityof10degreespersecond.Thekneejointanglewherepainor

discomfortisfeltwasconsideredastheendofthepainfreezone.

83

3.5.3MuscleSorenessAssessment

AlthoughDOMShasshowntobehighlysubjective,sorenessratingof

extensionhasshowntobesignificantlycorrelatedwithmaximalisometric

forcedecrementsandrelaxedjointangles(Nosakaetal.,2002).Forthe

assessment,whilerelaxedtheparticipant’slegwasslowlymovedtoamaximal

flexedpositionbytheexaminer.Subjectswerethenaskedtoreporttheirpain

sensationonaLikertscaleconsistingofaline(1-10)indicatingnopainatthe

leftendandseverpainindicatedonthefarright.Likertscaleswithfewer

responsecategorieshaveshowntolowerreliability,especiallytest-retest

reliability(refImpactofnumberofcategoriesandanchorlabelsoncoefficient

alphaandtest-retestLi-Jenweng(2004).Therefore,aLikertscaleconsistingof

options1-10forpainsensationwasemployed.Perceivedmusclesorenesswas

evaluatedatbaselineand24,48,and72hourspostexercise.

3.6ExerciseProtocol

Subjectswereinstructedtobeginwitha5-minutewarm-upona

stationarycycleergometeratanexerciseintensityof50wattsandmaintained

anRPMof50.Thecybexdynamometerwasthenusedtoinducemuscle

damageforbothgroupsandtheprotocolselectedhasbeenpreviouslyshown

toinduceasufficientdegreeofexerciseinducedmuscledamage(Aminian-Far,

Hadian,Olyaei,Talebian,&Bakhtiary,2011).Basedonkickingpreferencethe

dominantlegwasusedagainsttheleverarmoftheisokineticdynamometer.

84

Subjectswerestrappedintothedynamometerinaseatedpositionusing

chest,waist,andthighstraps.Theexercisebeganwiththeirkneefully

extended,(zerodegrees=fullextension)andendedwhenthekneereached

fullflexion.Sixsetsof10maximaleccentricvoluntarycontractionswere

performedwith3minutesofrestprovidedbetweensets.Theeccentricphase

velocitywassetto60degreespersecondandtheconcentricphaseto120

degreespersecond.Movementsbeganatakneeflexionof90degreesandthe

exercisewasinitiatedwhenfullextensionwasattained.Theparticipantswere

thenencouragedtoresistthecybexmotorarmmaximallythroughtheentire

rangeofmotionuntilfullflexionwasreachedandtorestduringtheconcentric

phaseastheirkneereturnedtotheextendedposition.Thisensuredthatonly

eccentricflexionoflegflexorswasperformed.

3.7DataAnalysis

Muscletorquedataispresentedasthemeanpeaktorque(Newton-

Metres).UsingIBMSPSSStatistics25(IBM®SPSSStatisticsSoftware,athree-

waymixedANOVAwasruntounderstandtheeffectsontreatmentA(Saffron

experimentalgroup)vsTreatmentB(placebo)overtimefollowingataxing

exerciseonmeanchangesinpeakkneeextensiontorqueandkneerangeof

motion.Perceivedmusclesorenessmeasureswerenotnormallydistributed

consequentlyaKruskal-Wallistestwasconductedtodeterminemedian

differencesbetweentheexperimentaltreatmentandtheplaceboovertime.

85

Iftreatmentxtimeinteractionswerefoundtobesignificantthent-

testswereusedtomakepairwisecomparisonsbetweentreatmentsateach

timepoint.Bonferronicorrectionsofpairwisecomparisonsbetween

treatmentandplacebotrialswasthenemployed.Analphalevelofp≤0.05

wasusedandconsideredsignificant.

86

4.0 Results

Thepurposeofthisresearchwastoinvestigatethepreventative

effectivenessof10-daysupplementationwith300mgofsaffronondelayed

onsetmusclesoreness,andselectedrelatedsymptoms(maximalpeaktorque

andchangesinkneerangeofmotion).Wehypothesizedthattherewillbeno

significantdifferencesfoundbetweentheplaceboandtreatmentgroups’

torque,kneerangeofmotion,andself-reportedmusclesoreness,pre-and24,

48,and72hourspostexercise.

4.1IsokineticTorque

Torquedatadistribution,presentedinFigure4.1.1andillustratedin

Figure4.1.2,meansandstandarddeviationsarepresentedinTable4.1.

Torquedatawasapproximatelynormallydistributed,exceptfor48-hourmale

control,p=.032,and48-hourfemalesaffrontrial,p=.031asassessedby

Shapiro-Wilk’stest(p>.05).Therewashomogeneityofvariancesasassessed

byLevene’stestforequalityofvariances.Mauchly’stextofsphericity

indicatedthattheassumptionofsphericityhadbeenmetfortreatment*time

interactions,𝑥)(5)=8.084,p=.153.Therewasnostatisticallysignificant

three-wayinteractionbetweentime,gender,andtreatment,(3,39)=.312,p=

.816.

87

TorqueDataDistribution

Figure4.1.1–TorqueDataDistribution-showstorquedatadistributionfor

experimentaltreatmentandplaceboatbaseline,24,48,and72hourspost

exerciseforbothmale(N=10)andfemaleparticipants(N=5).

88

Table4.1–DescriptiveTorqueData-displaysmeansandstandarddeviation

formaleandfemaletorquedatafortheexperimentaltreatmentandthe

placeboatbaseline,24,48and72hourspostexercise.

IsokineticTorque

Control

Treatment Female Mean Std.Dev Mean Std.Dev NBaseline 158.9 16.3 165.1 12.6 5Post24h 146.1 16.1 155.5 10.3 5Post48h 155.1 19.5 161.1 10.7 5Post72h 154.1 15.1 174.7 14.1 5Male

Baseline 249.2 13.1 245.7 9.0 10Post24h 234.4 7.9 250.2 9.9 10Post48h 240.1 10.3 249.1 9.0 10Post72h 238.5 10.9 251.1 8.7 10

89

PeakIsokineticTorque

Figure4.1.3–PeakIsokineticTorque-illustratesmeandifferencesinpeak

isokinetictorque(Nm)betweengender,theexperimentaltreatment,andthe

placeboovertime(mean±SD).

Therewerenostatisticallysignificantsimpletwo-wayinteractions

identified.Therewerenostatisticallysignificantmeandifferencesfound

betweentheexperimentalorcontrolgroupbaselinepeaktorquemeasures.

Moreover,therewerenostatisticallysignificantmeandifferencesfound

betweenthecontrolgrouptorquemeasureswhencomparedtothetreatment

atbaseline,24,48and72-hourmeasures.Therefore,weacceptthenull

hypothesisstatingthattherewillbenosignificantdifferencebetweenthe

90

placeboandtreatmentgroups’peakisokinetictorqueofthekneeextensorsat

aconstantspeedof60degreespersecondpre-and24,48,and72hourspost

exercise.

91

4.2RangeofMotion

ROMdatadistribution,presentedinFigure4.2.1andillustratedin

Figure4.2.2,meansandstandarddeviationsareshowninTable4.1.AllROM

datawereapproximatelynormallydistributed,exceptformale-48hour

measures(p=.01)asassessedbyShapiro-Wilk’stest(p>.05).Therewas

homogeneityofvariances,varianceforROMscores,asassessedbyLevene’s

testforequalityofvariances.

4.2.1-RangeofMotionDataDistribution

Figure4.2.1-RangeofMotionDataDistribution-showsROMdata

distributionforexperimentaltreatmentandplaceboatbaseline,24,48,and

72hourspostexerciseforbothmale(N=12)andfemaleparticipants(N=5).

92

Table4.2KneeROMDescriptiveData--displaysmeansandstandard

deviationforgenderandchangesinkneeROMdatafortheexperimental

treatmentandtheplaceboatbaseline,24,48and72hourspostexercise

KneeRangeofMotion

Control

Treatment Female Mean Std.Dev Mean Std.Dev NBaseline 144.6 4.0 142.7 3.1 5Post24h 144.0 4.0 141.7 3.8 5Post48h 143.5 4.6 142.7 3.5 5Post72h 143.9 4.2 142.8 3.5 5Male

Baseline 136.2 1.4 137.6 2.1 12Post24h 135.6 2.2 135.4 1.9 12Post48h 135.5 2.7 136.8 1.5 12Post72h 134.3 2.2 137.0 2.0 12

93

RangeofMotion

Figure4.2.3.RangeofMotion-illustratesmeandifferencesinkneeROM

betweengender,theexperimentaltreatment,andtheplaceboovertime

(mean±SD).

Forthe3-wayinteraction,Mauchly’stextofsphericityindicatedthat

theassumptionofsphericityhadbeenmet,𝑥)(5)=9.162,p=.104,for

treatmenttimeinteractions.Therewasnostatisticallysignificantthree-way

interactionwithintime,gender,andtreatment,F(3,45)=.149,p=.930.

Statisticalsignificanceofasimpletwo-wayinteractionwasacceptedata

Bonferroni-adjustedalphalevelof.025.Therewerenostatisticallysignificant

94

simpletwo-wayinteractionsidentified(p<.05).Therewerenostatistically

significantmeandifferencesfoundbetweentheexperimentalgroup’sbaseline

ROMmeasureswhencomparedtothetreatment.Moreover,therewereno

statisticallysignificantmeandifferencesfoundbetweenthemaleorfemale

controlgroup’sROMmeasureswhencomparedtotheexperimental

treatmentatbaseline,24,48and72-hourmeasures.Therefore,weacceptthe

nullhypothesisstatingthattherewillbenosignificantdifferencebetweenthe

placeboandtreatmentgroup’skneerangeofmotionpre-and24,48,and72

hourspostexercise.

95

4.3PerceivedMuscleSoreness

Reportedmusclesorenessdatadistribution,presentedinFigure3.1.1

andillustratedinFigure3.1.2;meansandstandarddeviationareshownin

Table3.1.Distributionsofpainmeasuresweresimilarforallgroups,as

assessedbyvisualinspectiontheboxplots.

4.3.1ReportedMuscleSorenessDataDistribution

Figure4.3.1–ReportedMuscleSorenessDataDistribution–showsreported

musclesorenessdatadistributionforexperimentaltreatmentandplaceboat

baseline,24,48,and72hourspostexerciseforbothmale(N=12)andfemale

participants(N=5).

96

Table4.3ReportedMuscleSorenessDescriptiveData-displaysmediansand

standarddeviationforgenderandchangesinmusclesorenessratingsforthe

experimentaltreatmentandtheplaceboatbaseline,24,48and72hours

postexercise.

MuscleSorenessScale1-10

Control

Treatment Female Mean Std.Dev Mean Std.Dev NBaseline 0 0 0 0 5Post24h 1.9 2.6 1 1.7 5Post48h 2.1 2. 1.6 1.8 5Post72h 1.8 1.8 0 0 5Male

Baseline 0 0 0 0 12Post24h 1.8 1.8 .8 .9 12Post48h 1.3 2.2 .3 .5 12Post72h .8 1.0 .3 .5 12

97

Figure4.3.2-PerceivedMuscleSoreness

Figure4.3.2.Perceivedmusclesoreness-illustratesmeandifferencesin

reportedquadricepsmusclesorenessbyaLikertscaleof1-10between

gender,theexperimentaltreatment,andtheplaceboovertime(median±

SD).

Medianpainscoresbetweentheexperimentaltreatmentandthe

controlwerenotstatisticallydifferentatbaseline,24or48-hourpost

measures.Howevermedianpainscoreswerestatisticallydifferentbetween

groupsat72hourspostexercise,𝑥)(3)=8.948,p=.03.Medianpainscores

werepainfreethefemaleexperimentaltrial(0)ascomparedtothefemale

98

controltrial(1.8)at72hourspostexercise.Pairwisecomparisonswere

performedusingDunn’s(1964)procedurewithabonferronicorrectionfor

multiplecomparisons.AdjustedP-valuesarepresented.Thisposthocanalysis

revealedstatisticallysignificantdifferencesinmedianVASpainscores

betweenfemaleexperimentaltrial(0)andthefemalecontroltrial(1.8)(p

=.043)group,butnotanyothergroupcombination.Therefore,werejectthe

nullhypothesisstatingthattherewillbenosignificantdifferencebetweenthe

placeboandtreatmentgroupsperceivedmusclesorenesspre-and24,48,and

72hourspostexercise.However,thisfindingonlypertainstofemale

participantsat72hourspostexercise.

Summaryofmainfindings:

• Therewerenostatisticallysignificantmeandifferencesfoundbetween

thetreatmentandplacebogroup’speaktorquemeasurespre-orpost-

exercise.

• Therewerenostatisticallysignificantmeandifferencesfoundbetween

thetreatmentandplacebogroupsROMmeasurespre-orpost-exercise.

• Therewasastatisticallysignificantdifferencefoundbetweenthe

treatmentandplacebogroupspainscores(p=.043)at72hourspost

99

exercise,butnotformaleparticipantsoranyothergroupcombination.

At72hourspostexercise,painscoreswerepainfreethefemale

experimentaltrial(0)ascomparedtothefemaleplacebotrial(1.8).

100

5.0DiscussionandConclusion

Currentpublishedresearchhasindicatedastrongpreventativeeffectof

10-daysupplementationwithsaffrononDOMSandrelatedsymptomsfor

maleuniversitystudents(Meamarbashi&Rajabi,2015).Toourknowledge,

thisisthefirststudytoexaminetheeffectsofsaffronwithbothmenand

womenonsymptomsofexerciseinducedmuscledamage.Accordingto(Reid,

2001),antioxidantactivitiescaneitherbenefitforceproductionorfatigue

preventioninhumansdirectlyorindirectly.Adirecteffectcouldbethe

reductionofmusclefatigueatthelevelofcontractilefunction.Indirecteffects

ofantioxidantactivitiesmayincludeenhancementofrecoveryfromtraining,

and/orthereductionofphysiologicalstressorsthatnegativelyaffecttraining

andrecovery.Therationaleforingestingpolyphenolsorpolyphenolrichfoods,

suchasSaffron,withrespecttoimprovementofforceproductionand

recoveryfollowingexerciseinducemusclesoreness(EIMD).

• Exercise-inducedexcessivefreeradicalproductionistoohighforthe

endogenousscavengingmechanisms.

• Musclemicro-damagecausesneutrophiloxidativebursts.

• Themyoglobinbreakdownproductsproduceferricacid.

• TheLipidmembraneandsurroundingproteinsdamagedbyoxidation

reactions.

101

However,emergingresearchsuggeststhatreducingcellularstressfrom

exercisemayinhibitendogenousadaptationstoregulartraining(Powers&

Jackson,2008).

Althoughitisnotwithinthescopeofthisstudytoidentifythefactorsinvolved

inthepreservationofstrengthorthereductionofmusclesoreness;our

hypothesiswasthatthecombinedanti-oxidantandanti-inflammatory

propertiesofSaffronsupplementationmaylessenoreliminateexercise

inducedDOMSand/orrelatedsymptoms.

Thisresearchwasinspiredbythepreviouslypublishedworksby

(Meamarbashi&Rajabi,2015)onsaffronprotectiveeffecttowardsDOMSand

relatedsymptoms;however,ourfindingsofarenotconsistentwithcurrent

publishedliterature.Wefoundnostatisticallysignificantdifferencesbetween

thesaffroninterventionandplacebogroupspeaktorquemeasuresat

baseline,24,48,or72hourspostexercise.However,ourresultsshowthat

therewerenoperformancereductionsinanyofthemaleexperimentaltrials

at24,48,and72hourspostexercisecomparedtobaseline,moreoverneither

malenorfemaleplacebotreatmentgroup’sfullyrecoveredforceoutputby72

hourspostexercise.Perhapsperformancemeasurestakenoverthecourseof

7daysinsteadofthreewouldhavebeenmoreusefulforestablishing

differencesinfullmusclerecovery.Thelackofsignificantdifferencesfound

betweenbaselineand24-hourpostexercisemeasuresindicatesthatthe

102

exerciseintensityselectedwasnotintenseenoughtoidentifydifferencesif

differencesdidexist.Thissametendencyisalsonoticedformaximumisotonic

forceoutputforthesaffrontreatmentgrouponlyinthepreviouslypublished

workonsaffronanditseffectsonDOMS.Thissuggeststhateithertheexercise

intensityprescribedtoinduceDOMSwaseithernotintenseenoughforthe

saffroninterventiongroupand/orthattheintensitieswerenotequivalentto

induceDOMSbetweentreatmentgroups.Alternatively,perhapssaffrondid

exertastrongprotectiveeffectonexerciseinducedDOMS.Inaddition,

similarlytothepreviouslypublishedworksbyMeamarbashietal,(2015)our

experimentalgroupshowedthehighestpeaktorquemeasuresforbothmale

andfemaleparticipantsat72hourspostexercise,anincreasefrombaselineby

2.2%and5.8%respectfully.Althoughthissmallincreaseinforceproductionat

72hourspostexerciseiswellwithinthestandarddeviationandcouldbedue

torandomerror,itcouldbeconceivablethatthesesmallincreaseswould

morelikelybeduetoimprovedmovementpatternsorneurallearning

adaptionsratherthananyperformanceenhancementbenefits.

Similarfindingshavebeenreportedonstrengthfollowing250mlof

pomegranatejuicesupplementationingestedtwicedailyfor8dayspriorto

eccentricexerciseofthekneeextensors,butnotelbowflexormuscles

(Tromboldetal.,2011).Theirresultsindicatedamild,acuteergogeniceffect

withpomegranatejuiceinonlytheelbowflexorsandnotthekneeextensor

103

musclesofresistance-trainedindividualsfollowingeccentricexercise.The

authorsattributedthisdiscrepancytoeither;thedifferentexerciseprotocols

employedbetweentheelbowflexorsandkneeextensors,ortheinherent

featureofthekneeextensorsthemselves(aphenomenonfromthedailyuse

ofquadricepsforambulation),whichprovidedprotectionfromweaknessand

soreness.Fortheelbowflexors,theexerciseprotocolconsistedofthreesets

of20maximaleccentricelbowextensionsoffullyresistingthefullrangeof

motionfromflexiontofullextension.Theprotocolusedfortheknee

extensorswasadjustedbecausekneeextensormaximaleccentrictorque

exceededthetorquelimitonthecybex.Consequently,theeccentricknee

protocolusedafixedresistancesetat110%oftheunilateralconcentric1RM.

Itmaybethattheinherentfeaturesofthekneeextensorsmayhaveprovided

someprotectiveeffecttowards(EIMD),andmorelikelythattheadjusted

exerciseforthekneeextensorswasnotasintenseasitwasfortheelbow

flexormuscles.However,becauseourstudyshowednosignificantdifferences

betweenthetreatmentorcontrolbaselineand24-hourpeakkneeextension

torquemeasuresitisinourcasethattheexercisestimulusselectedmaynot

havebeenintenseenoughmoresothananyinherentfeaturetheknee

extensorsmayhavefromambulation.

Therewerenostatisticallysignificantmeandifferencesfoundbetween

themaleorfemaleplaceboortreatmentgroupsknee(ROM)atbaseline,24,

104

48or72-hourpostexercise.ThefemaletreatmentgroupkneeROMfully

recoveredby48hourspostexercise,howeverthefemalecontrolgroupdid

notreachfullrecoveryby72hourspostexercise.Neitherthemaletreatment

norcontrolgroupreachedfullrecoveryofkneeROMby72hours’post-

exercise.Becausethemeasurementerrorofthegoniometermeasurements

arewithin3degreesandthedifferencesobservedarefractionalatbest,

thereforeanytrendsnoticedwouldmostlikelybeduetomeasurementerror.

Againthissuggeststhattheexerciseselectedmaynothavebeensufficiently

intensetoinducethelevelofDOMSneededtodetectdifferencesif

differencesdidexistregardlessofanyinherentfeatureofthekneeextensors

fromambulation.

Theprimaryfindingofthisstudywasthatat72hourspostexercise,the

femaleexperimentalgroupwaspainfreecomparedtothefemalecontroltrial;

amediandifferenceofself-reportedpainof1.8outofascaleof1–10.This

differencewastheonlysignificantdifferencereported(p=.043).Also,although

ourexercisestimuluswasdeemedlessthanidealtoinduceDOMStothe

degreethatwecanseesignificantdifferencesbetweenmeasuresoftorque

andROM,wedidhavehigherandmoreprolongedmusclesorenessratings

thanintheoriginalpublishedresearch.

105

5.1AnticipatedLimitations

Therewereanumberoflimitationswiththepresentresearch.Eachis

discussedinthefollowingsection.

Bioavailability-participantswereinstructedtotaketheirsupplements

attheirownaccordandthereforewerenotalltakenatthesametimepoints.

Moreover,notallmeasuresweretakenexactly24hourslaterduetotime

constrainsofparticipant’slifestyle.Therefore,itmayhavebeenusefulto

measureserumsaffronlevelsthroughoutthestudyasnutrientbioavailability

ofsubstanceswillvarybetweenindividuals.Evenwhenthebodyingestsa

knownsubstance,itisimpossibletoevaluatebioavailabilityeffectsbetween

individualswithoutregularbloodsampling,thusimposingthenecessityof

assessingsufficientindividualsforstatisticalanalysis.Polyphenol

bioavailabilityrangesfromtwoto20percentinanimalstudiesandistypically

10%orlessandhumans.Humansbeingmoregeneticallydiversethananimals

mayshowsimilardifferencesorevenperhapslarger(Manach,Williamson,

Morand,Scalbert,&Rémésy,2005).Forinstance,arangeof5–57%ofthe

naringin,aphytochemicalfoundingrapefruit,consumedwithgrapefruitjuice

wasfoundinhumanurinesamples(Fuhr&Kummert,1995).Fuhr(1995)

concludedthatchangesinthecompositionofthecolonicmicrofloracould

haveexplainedthelargeinterindividualvariationsinbioavailabily.Assuming

thatpolyphenolexposureinhumansissimilartoanimalstudies,regularblood

106

samplingshouldberecommendedtoestablishindividualbioavailabilitystatus

onsmallsamplesizeresearchstudies.Ifinterindividualvariationsin

bioavailabilityisnotfeasiblycontrolledfororestablished,thenalarger

populationpoolmaybenecessarytoidentifydifferencesifdifferencesdo

exist.

Inthepreviouslypublishedresearchonsaffronanditsclaimstowards

providingaprotectiveeffecttowardsDOMS,timeofingestionspecificallyset

at6pmforallparticipant(Meamarbashi&Rajabi,2015).Theirpretestand

posttestdatawerespecificallyobtainedbetween11to12ambeforethe

supplementationperiodandrepeated24,48,and72hoursaftereccentric

exercise.Bloodsamplestakenatregularintervalsduringsupplementation

wouldbeneededtoassessthekineticsofuptakeandeliminationofnutrient

supplements(Manachetal.,2005).Theeffectsofsaffron,ifany,aremostly

attributedtoitsheavypolyphenolcontent.In2005,Manachereviewed

polyphenol(flavanones)bioavailabilityextensivelyencompassing97studies

covering18differentpolyphenolsfromfood,juiceandoralsupplements.The

effectofthesecompoundsappearstobegreatlytiedtotheirtimetomaximal

plasmaconcentrationandithasbeenmadeclearthatthisconcentrationcould

varyfromonetoover6hourspostingestion.Notcontrollingforbioavailability

byregularbloodsamplinghasclearlimitationsinexercisestudies.Duetotime

constraintsandlimitedresources,wewereunabletodetermineindividual

107

bioavailabilityoranestimationoftimetomaximalbloodserumlevelsof

polyphenolsfromtheingestedsaffron.Itisunknownhowlongsaffronstaysin

thehumanphysiologyorifthebenefitsaremorefavorableduringpeak

polyphenolbloodserumlevels.However,incrocetin-administeredmice,

plasmacrocetinreacheditsmaxi-mumconcentrationwithin0.5hafterthe

doseandthendecreasedgradually(Asai,Nakano,Takahashi,2005).As

nutrientbioavailabilityofsubstanceswillvary,assessingbioavailabilityofpeak

polyphenolplasmaserumlevelswouldhelpclarifytheamountofthese

compoundsabsorbedandtheirtimeofpeakbloodserumlevelsfrom

ingestion.Itispossiblethatnone-responderssimplyarenotprocessingthe

substancesingestedasefficientlyandeffectivelyasothers.

Saffronquality-thequalitymayhavebeenalimitationasmany

intrinsicfactorscanaffectthequalitativeandquantitativeaccumulationof

biologicallyactivecompoundsproducedand/oraccumulatedinsaffron.Such

factorsaffectingthequalityofsaffronincludeandarenotlimitedto:

environmentalconditions,cultivationandfieldcollectionpractices,post

harvestinghandling,storage,manufacturing,adulteration,andeven

geographicalorigin(Kumaretal.,2008).Moreover,diagnosisofthreefamily

testsofcultivatedIranian,Grecian,andSpanishsaffronrevealedthatIranian

samplesarechemicallyverydifferentthantheGreekandSpanishsamples

(Zalacainetal.,2005).Toestablishsaffronqualitycertainrecommendations

108

aresetbyinternationalcommercialagreementasdeterminedbyISOguideline

3632(“ISO3632-1:2011,”).TheISOhassetaminimumrequirementUV-vis

spectrometryofthe3mainbioactivecompoundscrocin,picrocrocin,and

safranal,whichareresponsibleforsaffroncolor,flavorandaroma,

respectively(SirangRasaneh,2000).Howeverduetosomedoubtaboutthe

accuracyofthesafranalspectrometrymeasuresHadizadehetal,.(2006)

evaluatedandcomparedsamplesofsaffronusingtheISOmethodandhigh-

performanceliquidchromatography(HPLC).Theresultsindicatedthatcrocin

concentrationsintheanalyzedsaffronsampleswereclose,buttheISO

methodshowedtosignificantlyoverestimationoftheamountofsafranal.

Therefore,thequalityofthesaffronisregulatedbythecurrentevaluation

methodsusedtoquantifyitspotencyandquality.

Non-ChewableTablets-Ifthepillsweretobechewed,itwouldbe

easytoidentifythesaffrontabletsfromtheplacebo.Ifanyofthepillsare

chewed,themaybeabletoidentifybetweentreatments.Allparticipantswere

instructednottochewanyofthetreatmentpillsprovided.

Caffeineintake–Caffeineintakemayreduceforcedecrements

followingunaccustomedstrenuousexerciseandhasshowntoimprove

enduranceforthelowerbodymusculature(Davis&Green,2009;Warrenet

al.,1993).Caffeineactionsonperformanceenhancementandrecovery

requiremoreresearchasfewstudieshavebeenpublished.

109

5.2UnanticipatedLimitations

Therewereafewunanticipatedlimitationswiththepresentresearch.Eachis

discussedinthefollowingsectionbelow.

Exerciseprotocol–Duetothelackofsignificantdifferencesfound

betweenbaselineand24-hourpostexercisemeasures,itwassuspectedthat

theexerciseprotocolusedtoinduceDOMSwasnotintenseenoughtoinduce

thelevelofdamagetoseedifferencesifdifferencesdidexist.

Limitedsamplesize-Consideringthattherecruitmentof16maleand

16femaleparticipantswouldhavebeenidealforstatisticalanalysis,only13

malesand5femalesparticipated.Oftheserecruits,threemale’sdatawere

omittedfromtheisokinetictorqueanalysisandonefromboththeROMand

VASmeasures.Inadditiontohavingasmallsamplesize,largestandarderrors

wereconsequentlyformedfromsignificantdifferencesbetweenparticipant’s

torquemeasures.Thesedifferencesmayhavelimitedourabilitytoidentify

differencesbetweentreatments.Agoodmanystudiesonergogenic

nutraceuticalsupplementationresearchevaluatesmallnumbersofsubjects

andthereforeeitherthoughlargemeanchangesareobserved,resultsmaynot

reachstatisticalsignificance.Perhapsrecruitingparticipantsbetweentheages

of19-35wastooanarrowanagerangetoattainthedesiredparticipantpool.

Ifwehadbeenmoreliberalwithourageexclusioncriteria,wemayhavehada

largersamplesize.

110

ResearchProtocolOversight-Duetoaresearchprotocoloversight,

maximalisometrictorquemeasuresmaynothavebeenmeasuredatthesame

jointangle.Thejointanglewasapproximatelyfiveto10degreesoffdepending

onthedegreeofflexibilitybeyondorbelowtheangleoftheparticipant’sfull

kneeextension.Thisoversightlimitsourabilitytocompareisometrictorque

measuresconfidentlybutwebelievethattheeffectoftheoveralltorque

producedbetweenmeasureswouldbeminimalduetothecounterbalancing

designofthestudy.ThemaximalisometricmeasuresarelistedinAppendixD.

Lastly,asmallpilotstudywouldhavebeenhelpfulinreducingsomeofthese

limitationsandisrecommendedforfutureresearchinordertominimize

exerciseprotocolandequipmentprotocoloversights.

5.3Conclusion

Althoughwehadalimitedsamplesizeandunaccountedfor

bioavailabilityofsaffroningestion,preliminaryevidencesuggests10-day

supplementationofsaffronmayreducemusclesorenessfollowingeccentric

exerciseforfemales.However,nosignificantdifferenceswerefoundbetween

thesaffrontreatmentgrouporplacebobaselinecomparedto24,48,or72-

ourpost-exercisemeasures.Dueofthelackofsignificantdifferencesobserved

betweenbaselineand24-hourpost-exercisemeasures,itdifficulttodrawany

conclusions,astheexerciseintensityselectedwasdeemednotintenseenough

toidentifydifferencesifdifferencesdidexistbetweentreatments.Ourresults

111

indicatethattheeffectsofsaffroningestiononDOMSandrelatedsymptoms

are‘stilltooearlytotell’asmoreresearchisneededtoestablishstatistical

confidenceofeffectiveness.

112

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137

AppendixA–InvitationLetterandInformedConsent

InvitationLetterandInformedConsent

UniversityofNewBrunswick

Fredericton,N.B.

Study Title - The effect of 10-day supplementation of 300mg of saffron on

delayed onset muscle soreness and maximal isokinetic and isometric force

development 24, 48, and 72 hours post unaccustomed strenuous eccentric

exercise.

PrincipalInvestigator:BlairWarkMScESScandidateBlair.w@unb.ca

SupervisingInvestigators:UshaKurugantiPhDUkurugan@unb.ca

Purpose

Dear potential participants, we ask for your permission to enroll you as a

participantinaresearchstudyfocusedoninvestigatingthepreventativeeffect

ofsaffron(awidelyavailablecookingspice)ondelayedonsetmusclesoreness

(DOMS).Thebenefitsofthisresearchincludeimprovingourunderstandingof

how taking saffronmay help prevent and/ormanage delayed onsetmuscle

sorenessandtheconsequentialreducedperformancefactorsresultingfromit.

138

The research may provide valuable information for those concerned with

musculardiscomfortandpainthatcanlimitparticipationinsports,trainingand

strenuousactivity.

StudyContacts

If youhave anyquestions regarding the research goals, scheduling, or other

concerns,pleasecontactBlairWark,Blair.w@unb.ca,506-259-3737

Procedures

Asaparticipantwewillaskyoutotakeasaffrondietarysupplement(inthe

formofapill)onceadayfortendays(someparticipantsatsomepointduring

theexperimentwillbetakingaplaceboinstead).Ondaysevenofthisten-day

periodwewillaskyoutoexerciseunderourdirectioninsuchawayasto

inducemusclesoreness.Wewillthenmeasurethegradualreturnofstrength

andflexibilityondays8,9,and10,andcomparetherecoveryofparticipantsin

thetreatmentandplacebogroups.Afterthisyouwillbeallowedaneight-

weekbreak,andthentocomebackandrepeattheprocedure.

Duringeach10-daysupplementperiodyouwillbeprovidedwithatreatment

witnesslogandadietarylogsheet.Thetreatmentwitnesslogsheetand

dietarylogsheetwillbecompletedtothebestofyourabilityandreturnedto

139

theresearcher.Whenthesupplementistakenasignatureandcontact

informationofanyindividualofyourchoosingwillberequiredeverydayfor

eachandall10days.Thiswillnotonlyprovideawitnesstoverifythatthe

supplementwastaken,butwilllikelyhelpyouremembernottoforgettotake

it.Thedietarylogisabriefdescriptionofwhatyoueatanddrinkoverthe10-

dayperiod.

YouwillbeaskedtocometotheRichardCurryCentreontheUNBcampusin

Frederictonondays7,8,9,and10ofthedietarysupplementationperiod.You

willbeexpectedtowearshortsandgymclothesfortesting.Inaddition,you

willbeaskedtoattendonefamiliarizationsessionanytimebeforeday7of

supplementationwhereyouwillbeintroducedtotheequipmentand

procedures.TheCybexHUMACNorm(CSMI,USAInc.)isokinetic

dynamometerlocatedatUNBintheRichardCurryCentre,Frederictonwillbe

employedonday7ofsupplementationtoassessstrengthandtoinduce

musclesoreness.TheCybexwillalsobeemployedtoassessanystrength

reductionsonday8,9,and10ofsupplementation.Agoniometer(basicallya

giganticprotractor)willbeusedtoassesstherangeofmotionalongtheknee

jointbeforetheexerciseonday7andagainaftertheexerciseonday8,9,and

10.Moreover,onascalefrom1to10(1beingnopainand10beingsevere

pain)youwillrateyourdegreeofmusclepainonlyinthethighregionondays

140

7,8,9,and10.Datacollectionwillbeginafterparticipantssignandreadthe

informedconsentform,theWaterlooFootednessQuestionnaire,andParQ

plusform.TheWaterloofootednessquestionnaireisaseriesofquestionsthat

willbeusedtoidentifyyourdominantlegandaParQplusformisalsoa

questionnairedesignedtoassessyourabilitytoparticipateintheexercise

componentoftheresearch.

After you are strapped into the Cybex and ready for the first strength

assessmentyouwillpushagainsttheCybexmotorarmwithyourdominantleg

andallofyourmightwhileitismovingandwhenitisnotmovingatall.During

thestrengthassessmentwhiletheCybexmotorarmismovingandreachesthe

speedlimitsetbytheresearcheranyadditionalforceappliedwillberecorded.

This is thesamesituationwhenyouwillbeasked topushagainst theCybex

motorarmwhenitisprogrammednottomove.Alloftheforceappliedwhenit

is not able to move will be measured and recorded. The exercise protocol

intendedtoinducedelayedonsetmusclesorenesswillalsoinvolveyoupushing

yourshinagainsttheCybexmotorarmwhileatthesametimethemachinewill

pushback.Thiswillallowyourthighmusclestolengthenwhiletheyareunder

tension(calledaneccentricmusclecontraction)foratotalof60reps(6setsof

tenreps).Aftertheexerciseprotocolstrengthmeasureswillbetakenagainfor

comparison between before and after exercise. All selected measures

141

(maximum strengthmeasures, knee range ofmotion, and perceivedmuscle

soreness,ifany)willbetakenbeforetheexerciseintendedtoinducedelayed

onsetmusclesorenessonday7andonday8,9,and10forcomparison.Also

youwillbeaskedtonottoconsumecaffeinefor2hoursbeforetesting(and

preferablynoneatall).Afterthefirst10dayperiodofsupplementationan8

week break will be provided where you will be given the other treatment

(treatmentAorB)thatyouweredidnottakebeforeandtheprocessisrepeated

oncemore.

Table5–illustratestheoutlineoftheresearchdesigninvolving10daysofsupplementation,

theexerciseprotocol,andselectedindirectmeasuresofDOMS.

Daysonethrough

sixof300mgof

saffron

supplementationor

placebo.

-Beforedata

collectionbegins,

one-familiarization

sessionsofprotocol

expectationsand

equipmentsetup

Baselinemeasures

ofthefollowing

taken:

ROM

MuscleSoreness

Maxstrength(peak

torque)

*24hourspost

exercise

Measuresof:

ROM

*48hours

postexercise

Measuresof:

ROM

*72hours

postexercise

Measuresof:

ROM

142

willtakeplace

duringthistime.

Day1-6

*ExerciseProtocol

Execution

Postexercise,peak

torquemeasures

Day7

Muscle

Soreness

Maxstrength

Day8

Muscle

Soreness

Maxstrength

Day9

Muscle

Soreness

Maxstrength

Day10

Figure1showstheCybexduringkneeflexionasshownintheimagetotheleftandknee

extensionasshownintheimagetothefarright.

ExclusionCriteria

Onlyhealthymaleandfemaleparticipantsbetweentheagesof19and35will

berecruited.Thosewhoadmittoparticipationinresistancetrainingwithinthe

last3monthsand/ordiagnosedwithdiabetesoranyothercirculatorydisorder

willbeexcludedfromthestudy.Lastly,anypastorpresentinjurythatwould

remotelycauseanyrisktotheparticipantiscauseforexclusion.

143

Costs

Thereisnodirectcosttoyouforparticipationexceptforyourtimeandyour

duediligence.Ifforanyreasonyoufeelyouhaveexperiencedextracostsfor

participationthatareunwarranted,pleasecontacttheresearcher.

RisksandDiscomforts

Theeccentricexercises,whichyouwillbeaskedtocompleteontheseventh

dayofthisproject,aredesignedtomakethekneeextensormusclesinyour

dominantlegquitesore.Mostparticipantswilltemporarilylose20%oftheir

normalmuscleuse,anduptoa40%lossmaybeexperiencedinsomecases.

Youcanexpecttoexperiencepainandsorenessforuptofourtofivedays

afterwards,anditmaybeseventotendaysbeforeyouregainfullmuscle

strength.

Althoughthereissomeuncertaintyaboutthesafetyofsaffronwhen

consumedforlongperiodsorathigherdoses,thesaffronpillsyouareasked

totakeinthisprojectareconsideredtobesafeformedicinalconsumption.

Saffronisawidelyusedcookingspicethatisharvestedfromthecrocussativus

flowerandiscommoninmanydiets.Itisalsousedasaremedyagainst

variousconditionsintraditionalmedicine,andhasbeenshowntobeeffective

144

againsthighbloodpressure,depression,andtissueinflammation.Inseveral

recentstudiesusingsaffronorsaffron-extractsagainstdepression,some

participantsreportedadverseeventsduringthestudywhichincludedanxiety,

decreasedappetite,and,headache.Butthenumberofsuchreportswasnot

statisticallysignificant,norstatisticallydifferentfromthenumberofadverse

eventsreportedbyparticipantsinthecontrolgroupofthesestudies,i.e.those

whotooksugarpillsorsomeotherplaceboratherthansaffron.Arecent

scientificreviewofthesafetyofthemedicaluseofsaffronandsaffronextracts

concludedthat“dailydosesofupto1.5g[perday]ofsaffronaregenerally

consideredsafe.”Toxiceffectsarereportedwithdosesof5gandmore.The

dailydoseusedinthisstudyis300mg.

Ifyouconsenttoparticipateinthisproject,therearecertainsafety

considerations,whichyoushouldbeawareof:

• Treatthesaffronpillsasyouwouldtreatanyothermedicine:they

shouldbekeptoutofthehandsofchildren,andusersshouldtakecare

nottoexceedthedailydosageassignedbytheresearchteam.

• Womenwhoarepregnantshouldnottakesaffronorparticipateinthis

study.

• Individualswithfoodallergiesshouldnotparticipateinthisstudy.

145

• Anychangeinyourhealthduringorimmediatelyfollowingtheproject

shouldbepromptlyreportedtotheresearchteam.

Ifyouareuncertainaboutwhetherconsumingsaffron,asamedicineissafe

foryouasanindividual,discussthiswiththeresearcherbeforegivingyour

consenttobeenrolled.

PrivacyandConfidentiality

Your participation is confidential. Study informationwill be kept in a secure

locationattheUniversityofNewBrunswick.Allpersonalinformationgathered

forthisstudywillbekeptsecuretoprotecthisorherprivacyandwillnotbe

shared with anyone. Data collected and shared will be referenced by

alphanumeric code and will be kept in a secure location. All personal

informationlinkingparticipantstotheirdatawillbedestroyedoneyearafter

the completion of the study. De-identified information gathered from

participantswillbeusedandsharedwithforresearchpurposesonly.Theresults

ofthestudymaybepublishedorpresentedatprofessionalacademiclevel.

Ifyouwishtoreceivefeedbackoftheresultsofthisstudyyoucanleaveeither

youremailaddressormailingaddressandpermissiontosendyouasummary

oftheresearchfindings.Takingpartinthestudyisyourdecision.Youmayalso

quitbeinginthestudyatanytime.Ifyouhaveanyquestionsorconcernsyou

146

are encouraged to contact the Principal Investigator (Blair.w@unb.ca),

Supervisor(ukurugan@unb.ca).Ifyouhaveanyquestionsaboutyourrightsas

aresearchparticipant,youmaycontacttheResearchEthicsBoardchair(Steven

Turner) viaemail atETHICS@unb.ca. Thisprojecthasbeen reviewedby the

ResearchEthicsBoardoftheUniversityofNewBrunswickandisonfileasREB

2016-127.

Thankyouforyourconsideration.Ifyouwouldliketoparticipate,pleaseemail

meatBlair.w@unb.ca.

Withkindregards,

BlairWark

147

ConsentRespondentAgreement

I,theundersigned,doherebyacknowledge:

• I understand the purpose, procedures and risks involved, including theexerciseprotocoltoinducemuscledamage.

• AlsoanyquestionsorclarificationsIhaverequestedhavebeenexplainedtomysatisfactionbytheresearcher.

• Theresearchhasbeenexplainedtomeinaclearandconcisemanner.• Ivoluntarilyconsenttoparticipate.• Ihavehadanopportunityformyquestionstobeanswered.• IacknowledgethatImayrefusetoparticipateortostopmyparticipationin

theresearchatanytime.• I understand that in addition to the required dietary log and witness

supplementationformIwillberequiredtoparticipateinthisexperimentforapproximately3.5hoursovera4-dayperiodthatwillberepeatedagain6-8weekslateraccumulatingapproximatelyatotalof7hours.

• IunderstandthatifIhaveanyfurtherquestionsaboutthisresearchproject,I may contact the graduate student facilitating the researchBlair.w@unb.ca or his faculty advisor, Usha Kuruganti atukurughan@unb.ca.

Name(printname) Signature

Date

Witness(printname) Signature Date

□Iwishtoreceiveasummaryofresearchfindingsuponcompletion

Email/Address: ________-

______________________________________________

Haveyouengagedinregularlowerbodyresistancetrainingwithinthelast3

148

months?

YesNo

Areyouexpectingachild(pregnant)?Doyouhaveanyfoodallergies?

YesNoYesN

149

AppendixB–WaterlooFootednessQuestionnaire-Revised

WaterlooFootednessQuestionnaire–RevisedParticipant:_______________________________ Age:______ Sex:MFInstructions:Answereachofthefollowingquestionsasbestyoucan.Ifyoualwaysuseonefoottoperformthedescribedactivity,circleRaorLa(forrightalwaysorleftalways).IfyouusuallyuseonefootcircleRuorLu,asappropriate.Ifyouusebothfeetequallyoften,circleEq.Pleasedonotsimplycircleoneanswerforallquestions,butimagineyourselfperformingeachactivityinturn,andthenmarktheappropriateanswer.Ifnecessary,stopandpantomimetheactivity.1. Whichfootwouldyouusetokickastationaryballata

targetstraightinfrontofyou?LaLuEqRuRa

2. Ifyouhadtostandononefoot,whichfootwoulditbe?

LaLuEqRuRa

3. Whichfootwouldyouusetosmoothsandatthebeach?

LaLuEqRuRa

4. Ifyouhadtostepupontoachair,whichfootwouldyouplaceonthechairfirst?

LaLuEqRuRa

5. Whichfootwouldyouusetostomponafastmovingbug?

LaLuEqRuRa

6. Ifyouweretobalanceononefootonarailwaytrack,whichfootwouldyouuse?

LaLuEqRuRa

7. Ifyouwantedtopickupamarblewithyourtoes,whichfootwouldyouuse?

LaLuEqRuRa

8. Ifyouhadtohopononefoot,whichfootwouldyouuse?

LaLuEqRuRa

9. Whichfootwouldyouusetohelppushashovelintotheground?

LaLuEqRuRa

10. Duringrelaxedstanding,peopleinitiallyputmostoftheirweightononefoot,leavingtheotherlegslightlybent.Whichfootdoyouputmostofyourweightonfirst?

LaLuEqRuRa

11. Isthereanyreason(i.e.injury)whyyouhavechangedyourfootpreferenceforanyoftheaboveactivities?

YESNO(circleone)

12. Haveyoueverbeengivenspecialtrainingorencouragementtouseaparticularfootforcertainactivities?

YESNO(circleone)

IfyouhaveansweredYESforeitherquestion11or12,pleaseexplain:(usebackifnecessary)

150

AppendixC–WitnessTreatmentForm

Iherebyacknowledgethatthefollowinginformationprovidedisaccurateandcomplete

ParticipantSignature

Note:Thisformmustbecompleted,signed,andsubmittedtotheresearcherfollowingcompletion.Witnessesmustbeoftheageofmajorityandindependentoftheappraiser.Contactinformationofthewitnessmayberequestedfollowingformcompletion.Thankyouforyourparticipation.

WitnessTreatmentForm

ResearchProject-Theeffectof10-daysupplementationof300mgofsaffronondelayedonsetmusclesorenessandmaximalisotonicandisometricforce

development24,48,and72hourspostunaccustomedstrenuouseccentricexercise.

ParticipantSignature WitnessSignature Date/Time Comments

151

AppendixD–MaximalIsometricTorqueDataMeasures

Maximalisometricmeasuresformaleandfemaleparticipantsarelocated

below.

MaxIsometricToque(Nm)(ExperimentalTreatment)

Participant Baseline Zero 24hour 48hour 72hour1 264.7 252.7 278.2 264.4 246.82 189.0 220.8 237.1 233.4 234.43 347.5 293.3 333.4 334.4 326.74 221.5 226.0 304.5 302.4 245.05 354.5 306.4 299.5 321.0 328.46 315.8 287.5 380.5 404.3 369.67 279.2 259.4 294.2 302.2 329.88 277.0 249.9 253.1 276.3 299.79 296.7 266.5 281.0 286.3 344.010 314.2 234.2 184.1 275.0 299.911 344.5 323.4 335.0 347.3 334.212 367.8 324.3 357.9 387.6 426.913 237.3 195.1 192.7 230.5 224.8

Average 291.3 264.6 287.0 305.0 308.5MaxIsometricTorque

(Control)Participant Baseline Zero 24hour 48hour 72hour

1 226.9 203.6 245.6 270.7 277.82 202.8 215.8 273.6 183.1 NA3 345.7 265.6 282.6 315.2 314.34 252.8 209.3 229.8 252.8 228.15 370.4 325.6 327.4 349.2 341.06 306.6 294.9 338.7 321.9 355.37 367.8 221.6 372.5 NA 329.88 238.9 248.7 253.0 258.8 242.59 354.9 239.0 344.2 367.4 398.910 329.2 275.0 325.6 283.7 300.011 371.9 358.7 340.4 335.0 314.912 362.3 344.5 432.1 367.6 373.213 222.6 193.5 244.5 209.5 220.0

Average 304.1 261.2 308.4 292.9 308.0

152

MaxIsometricToque(FemaleExperimentalTreatment)

Participants Baseline Zero 24hour 48hour 72hour1 154.4 121.4 127.5 141.4 140.72 180.7 179.8 210.3 182.5 175.53 213.9 143.3 136.9 155.3 182.14 269.6 223.8 233.4 253.8 272.65 152.5 140.4 145.4 196.1 191.0

Average 175.4 161.7 170.6 185.8 192.4MaxIsometricTorque

(FemaleControl)Participant Baseline Zero 24hour 48hour 72hour

1 128.1 143.9 136.9 119.6 127.22 124.8 115.1 142.7 186.4 158.03 163.1 140.1 143.9 147.6 153.54 269.6 247.9 306.8 304.4 270.55 151.3 155.7 175.5 178.7 175.5

Average 167.3 160.5 181.2 187.3 176.9

Measureshighlightedwerecompletedatthesamejointanglewhereasthe

none-highlightedmeasuresmayhavebeentakenatdifferentjointangles.The

jointanglewasapproximately5to15degreesoffdependingonthedegreeof

flexibilityaboveandbeyondorbelowtheangleoftheparticipant’sknee

extension.

CurriculumVitae

BlairJeremyWark

UNBFredericton(2008-2013),BachelorofScienceinKinesiology(BSKIN)

Publications:None

ConferencePresentations:None