The effects of a multi-ingredient dietary supplement on body composition, adipokines, blood lipids, and metabolic health...

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Title:
The effects of a multi-ingredient dietary supplement on body composition, adipokines, blood lipids, and metabolic health in overweight and obese men and women: a randomized controlled trial
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English
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Ormsbee, Michael J.
Rawal, Shweta R.
Baur, Damiel A.
Kinsey, Amber W.
Elam, Marcus L.
Spicer, Maria T.
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Bi Med Central ( JISSN, Journal of the international Society of Sports Nutrition)
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Abstract:
Background: The present study investigated the effects of a multi-ingredient dietary supplement (MIDS) containing caffeine, conjugated linoleic acid (CLA), green tea, and branched-chain amino acids (BCAA) taken for 8 weeks on body composition, blood lipid profile, glucose, insulin, adiponectin, leptin, and high-sensitivity C-reactive protein (hs-CRP) in overweight and obese men and women. Methods: Twenty-two participants completed the study (PL, n = 11; 7 women, 4 men; age, 34 ± 3.5 years; height, 169.2 ± 3.3 cm; body mass, 96.9 ± 6.8 kg; BMI, 34.1 ± 1.8 kg/m2; MIDS, n = 11; 9 women, 2 men; age, 36 ± 3.4 years; height, 173.2 ± 2.9 cm; body mass, 91.9 ± 5.6 kg; BMI, 30.0 ± 1.5 kg/m2). Participants were randomly assigned and stratified by body fat percentage to two groups: 1) a soybean oil placebo (PL) or 2) MIDS. Each group consumed two pills with breakfast and two pills with lunch. Body composition and android fat, waist and hip circumferences, blood pressure and heart rate were measured at baseline and after 8 weeks of supplementation. Results: There were no significant changes for any of the variables of body composition. Feelings of hunger were significantly higher in MIDS versus PL with no changes observed in satiety or desire to eat. Heart rate and blood pressure were unaltered in MIDS after 8 weeks of supplementation. Furthermore, lipid profile, food intake, mood state variables, fasting blood glucose, and endocrine markers did not significantly change regardless of group. Conclusion: MIDS intake does not appear to alter body composition or markers of cardiovascular health versus PL. Moreover, MIDS may actually increase feelings of hunger versus PL. Keywords: Green tea, Caffeine, Conjugated linoleic acid, Branched chain amino acids, Body composition, Obesity
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Ormsbee et al. Journal of the International Society of Sports Nutrition 2014, 11:37 http://www.jissn.com/content/11/1/37; Pages 1-10
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doi:10.1186/1550-2783-11-37 Cite this article as: Ormsbee et al.: The effects of a multi-ingredient dietary supplement on body composition, adipokines, blood lipids, and metabolic health in overweight and obese men and women: a randomized controlled trial. Journal of the International Society of Sports Nutrition 2014 11:37.

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© 2014 Ormsbee et al.; licensee BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly credited. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.
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RESEARCHARTICLEOpenAccessTheeffectsofamulti-ingredientdietary supplementonbodycomposition,adipokines, bloodlipids,andmetabolichealthinoverweight andobesemenandwomen:arandomized controlledtrialMichaelJOrmsbee1,2,3*,ShwetaRRawal1,DanielABaur1,AmberWKinsey1,MarcusLElam1,MariaTSpicer1, NicholasTFischer1,TakudzwaAMadzima1andDDavidThomas1AbstractBackground: Thepresentstudyinvestigatedtheeffectsofamulti-ingredientdietarysupplement(MIDS)containing caffeine,conjugatedlinoleicacid(CLA),greentea,andbranched-chainaminoacids(BCAA)takenfor8weekson bodycomposition,bloodlipidprofile,glucose,insulin,adiponectin,leptin,andhigh-sensitivityC-reactiveprotein (hs-CRP)inoverweightandobesemenandwomen. Methods: Twenty-twoparticipantscompletedthestudy(PL,n=11;7women,4men;age,343.5years;height, 169.23.3cm;bodymass,96.96.8kg;BMI,34.11.8kg/m2;MIDS,n=11;9women,2men;age,363.4years; height,173.22.9cm;bodymass,91.95.6kg;BMI,30.01.5kg/m2).Participantswererandomlyassignedand stratifiedbybodyfatpercentagetotwogroups:1)asoybeanoilplacebo(PL)or2)MIDS.Eachgroupconsumed twopillswithbreakfastandtwopillswithlunch.Bodycompositionandandroidfat,waistandhipcircumferences, bloodpressureandheartrateweremeasuredatbaselineandafter8weeksofsupplementation. Results: Therewerenosignificantchangesforanyofthevariablesofbodycomposition.Feelingsofhungerwere significantlyhigherinMIDSversusPLwithnochangesobservedinsatietyordesiretoeat.Heartrateandblood pressurewereunalteredinMIDSafter8weeksofsupplementation.Furthermore,lipidprofile,foodintake,mood statevariables,fastingbloodglucose,andendocrinemarkersdidnotsignificantlychangeregardlessofgroup. Conclusion: MIDSintakedoesnotappeartoalterbodycompositionormarkersofcardiovascularhealthversusPL. Moreover,MIDSmayactuallyincreasefeelingsofhungerversusPL. Keywords: Greentea,Caffeine,Conjugatedlinoleicacid,Branchedchainaminoacids,Bodycomposition,ObesityBackgroundObesityisagrowingtrendintheUnitedStateswith currentestimatessuggestingprevalenceashighas35% amongadults[1].Thesehighratespotentiateasevere healthcrisisasobesityincreasesthelikelihoodofdevelopingchronicdiseaseswhichincludehypertension,insulin resistance,type2diabetesmellitus(T2DM),arteriosclerosis,coronaryheartdisease,andmetabolicsyndrome[2]. Therefore,findingsafeandeffectivemethodsforreducing bodyweightinobeseindividualsisessential. Reducingbodyweightrequiresmanipulationoftheenergybalanceequationtoproduceenergydeficits.Thiscan beaccomplishedthroughdietandexercise,pharmacologicalinterventions,orsurgicalmeans.However,eachof thesemethodscomeswithdisadvantages.Forinstance, manydietandexerciselifestyleinterventionssufferfroma lackoflong-term( 1yr)adherence[3].Furthermore, *Correspondence: mormsbee@fsu.edu1DepartmentofNutrition,FoodandExerciseSciences,FloridaState University,Tallahassee,FL32306,USA2UniversityofKwaZulu-Natal,Durban,SouthAfrica Fulllistofauthorinformationisavailableattheendofthearticle 2014Ormsbeeetal.;licenseeBioMedCentralLtd.ThisisanOpenAccessarticledistributedunderthetermsoftheCreative CommonsAttributionLicense(http://creativecommons.org/licenses/by/4.0),whichpermitsunrestricteduse,distribution,and reproductioninanymedium,providedtheoriginalworkisproperlycredited.TheCreativeCommonsPublicDomain Dedicationwaiver(http://creativecommons.org/publicdomain/zero/1.0/)appliestothedatamadeavailableinthisarticle, unlessotherwisestated.Ormsbee etal.JournaloftheInternationalSocietyofSportsNutrition 2014, 11 :37 http://www.jissn.com/content/11/1/37

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pharmacologicaland/orsurgicalmeanstoreducebody weightaretypicallyexpensiveandaresometimesaccompaniedbypotentiallyunpleasantand/ordangerousside effects[4,5].Assuch,considerationofalternativeweight lossmethodsiswarranted. Theconsumptionofnaturalingredientsand/ordietary supplementsmayprovideasafeandeffectivemeansto induceweightlossandimproveoverallhealth.Indeed, recentevidencesuggeststhatconsumptionofcertain multi-ingredientdietarysupplementsmayimprovebody compositionandmoodstate[6,7].Thesupplementsutilizedinthesestudiescontaineduniqueproprietary blendscontainingsuchingredientsascaffeineandgreen teaextract.Theseingredientshavebeenstudiedextensivelyandarenowrecognizedaspotentialmodulatorsof bodyweight,composition,adipokines,andmetabolic health[8,9].Beneficialeffectsofconsuminggreenteaor caffeine,respectively,arelikelyaresultofinhibiteddegradationofcatecholamines(epinephrineandnorepinephrine)orcyclicaminomonophospates(cAMP)thereby enhancingthermogenesisandpromotinglipolysis[10,11]. Ofinterest,thelipolyticeffectsreportedwithgreentea andcaffeineconsumptionmaybetheresultofasynergisticeffectasnotedinarecentmeta-analysis[12]. Supplementationwithconjugated-linoleicacid(CLA) andbranched-chainaminoacids(BCAA)mayalsoprovide weightlossbenefits[13,14].TheseCLA-derivedeffectsmay bearesultofenhanced -oxidationviastimulationofenzymesresponsiblefortransportoflipidsintothemitochondria(i.e.carnitinepalmitoyltransferase[CPT1])[15],or throughinhibitionofadipocytedifferentiation[16].BCAA mayimprovebodycompositionbyenhancingproteinsynthesis,whichmaintainsorincreasesleanmass[17].While theefficacyoftheseingredientshasbeenextensivelystudiedindividually,thereisalackofresearchontheimpactof combiningtheseingredientso nbodyweight,composition andothermarkersofhealth. Furthermore,combiningCLA andBCAAwithgreenteaextractsandcaffeinehasalsoyet tobestudied. Therefore,thepurposeofthepresentstudywasto examinetheeffectsofamulti-ingredientdietarysupplement(MIDS)containingaproprietaryblendofgreentea extracts,caffeine,CLA,andBCAAonbodycomposition, bloodlipidprofile,glucose,insulin,adiponectin,leptin, andhigh-sensitivityC-reactiveprotein(hs-CRP)concentrationsinoverweightandobesemenandwomen.MethodsParticipantsThirty-fourinactive(<2times/weekofplannedphysical activityfornomorethan60minutespersession)overweightorobese(BMIof25to47kg/m2)butotherwise healthymenandwomen,ages18 – 50yearsoldwererecruitedfromTallahassee,Floridaandsurroundingareas. Priortoparticipation,eachparticipantprovidedawrittenconsentandcompletedabriefquestionnaireregardinghisorhermedicalandexercisetraininghistory. Participantswereexcludediftheywerephysicallyactive (>2times/weekofplannedphysicalactivityfor>60minutes),iftheyhaduncontrolledhypertension(BP>140/ 90mmHg),iftheyhadhighlow-densitylipoproteincholesterol(LDL>160mg/dL)oriftheytookcholesterol medication.Inaddition,thosediagnosedwithcardiovasculardisease,stroke,diabetes,thyroidorkidneydysfunction,andsmokers(>5cigarettesperweek)wereexcluded. Thosewhoconsumedanydietarysupplementsintended toalterbodycompositionandbodyweightwereexcluded. Inaddition,thosewhohadanyallergiestosoy,wheat,and grainproductsthatwouldcauseahealthproblemwereexcluded.Thisstudywasapprove dbyFloridaStateUniversity InstitutionalReviewBoard.StudydesignThiswasarandomized,double-blinded,placebo-controlled studywithtwotreatmentgroups.Theparticipantswere stratifiedbasedonbodyfatpercentageandassignedtoa placebogroup(PL;soybeanoil)oranisocaloricmultiingredientdietarysupplementgroup(MIDS;Suarez CorporationIndustries,Canton,Ohio;10calories;1gfat, 99mgcaffeine,1510mgofaproprietaryblendof:green tea,CLA,andBCAAper2-pillserving[2servings=4 pills/day]).Thegreenteawasstandardizedfor45%epigallocatechingallateand90%polyphenols.Bothgroups ingested2identicalpillswithbothbreakfastandlunch 7daysperweekfor8weeks. Alllaboratoryprocedureswereconductedfollowingan 8 – 10hovernightfastanda24habstinencefromcaffeine,alcoholintakeandanyintensephysicalactivity. Additionally,participantsabstainedfromconsumingtheir assignedtreatmentontestingdays.Everyvisitwascompletedbetween7amand10am.AnthropometricsHeightandbodymassweremeasuredusingawall-mounted stadiometerandadigitalscale(SECA,Birmingham,UK) every2weeksunderidenticalconditions(shorts,t-shirt)for eachsubject.Waistandhipcircumferenceswereobtained atbaselineandafter8weeksusingastandardtapemeasure(withstraingauge)atthemaximumareaaroundthe waistlineandaroundthelargestcomponentofthegluteusmaximus.Allcircumferencemeasurementswere conductedbythesameresearcher. Totalandregionalbodycompositionwasdeterminedby dualenergyX-rayabsorptiometry(iDXA;GELunar, Madison,WI)withparticipantsinthesupineposition. Totalbodyadipositywasexpressedaspercentbodyfat (%BF).Abdominaladipositywasdeterminedbycreatingregionsofinterest(ROI)fortheabdomen(region1)Ormsbee etal.JournaloftheInternationalSocietyofSportsNutrition 2014, 11 :37Page2of10 http://www.jissn.com/content/11/1/37

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usingtheROIoptionwithinthemanualanalysismenu oftheiDXAsoftwareandincludedtheareajustbelow thelastribtojustabovetheiliaccrest,asdescribedpreviously[18].Abdominaladip ositywasexpressedaspercentandroidfat(%androidfat).Acertifiedradiologist technicianperformedalliDXAanalyses. Waistcircumferencesweremeasuredatthemaximum circumferencearoundtheumbilicalpoint.Althoughthisis notthestandardmeasurementmethodforwaistcircumference,theabdominalobesityofmanyoftheparticipants preventedtheuseofstandardwaistmeasurementtechniques(i.e.1 – 2cmbelowthelastribtypicallycorrespondingwiththenarrowestpartofthewaist).Measurements weretakenbythesameresearchertominimizeerrors.HeartrateandbloodpressureRestingheartrateandbloodpressureweremeasuredat0, 2,4,6,and8weeks.Theparticipantswereinstructedto restinaseatedposition,withbothfeetflatonthefloorfor 5minutesbeforemeasurementsweretaken.Restingblood pressureswereobtainedbythesameinvestigatoronthe samearmusinganappropriatelysizedcuffandasphygmomanometer(AmericanDiagnosticCorp.,Hauppauge, NY).Restingheartratewasobtainedfromaradialpulse. Tworeadingsweretakenforbloodpressuresandresting heartratewith1minintervalsbetweeneachreading.The meanofthetworeadingswereusedforstatisticalanalysis.Questionnaires:dietaryintake,physicalactivity,hunger, andmood-stateTwo-daydietlogswerecompletedatweeks0,4,and8 toquantifycalorieandmacronutrientintake.Theparticipantswereaskedtomaintaintheirfoodintakethroughoutthedurationofthestudy.TheFoodProcessor software(version10.9ESHAResearchSalem,OR,USA) wasusedtoanalyzethefoodlogs. Participantswereaskedtonotchangetheirphysical activityduringinterventionperiod.Physicalactivitylogs completedbyeachparticipantwerecollectedatweeks0, 4and8.Adverseeventsformswerecollectedandrecordedatweeks2,4,6and8byresearchstaff. Moodstatewasaccessedviathecompletionofa65questionmoodstatequestionnairewhichassessedmood stateatweeks0,4and8.Thiswasasimplelikertscale(0, very – 4,notatall)questionnaire.Hungerwasassessedat thesametimepointsusingasimplevisualanalogscale (VAS)(1to100mm).Participantswereinstructedtoplace amarkonthe100mmlinetoindicatetheirlevelsofhunger,satiety,anddesiretoeat.Amarkat0indicateda completelackofhunger,satiety,ordesiretoeatanda markat100indicatedextremehunger,satiety,ordesireto eatforeachVAS,respectively.Foreachofthethreemeasures(hunger,satiety,anddesiretoeat),thedegreein whicheachsensationwasfeltwasquantifiedbymeasuring howfarthemarkwasfromthe0mmmark.Forthis measurement,astandardmillimeterrulerwasusedand allscoreswerecomputedbythesameinvestigator.VAS scaleswerecompletedatbaseline(week0)andpostintervention(week8)inafastedstate.Hormones,bloodlipidsandbloodglucoseAvenousbloodsample(20ml)wasobtainedfromtheantecubitalveinatbaselineandafter8weeksofsupplementation (post).Wholebloodwasusedtomeasuretotalcholesterol (TC),high-densitylipoproteincholesterol(HDL-C),low densitylipoproteincholestero l(LDL-C),triglycerides(TRG), andglucoseconcentrationsusingtheCholestechLDXblood analysissystem(Hayward,CA).Inter-assaycoefficientsof variationwere2.1%,4.0%,4.1%,4.7%,and2.3%forTC, HDL,LDL,TRG,andglucose,respectively.Remaining sampleswerecentrifuged(IECCL3RMultispeedCentrifuge, ThermoElectronCorporation,NeedhamHeights,MA)for 15minutesat3500rpmat4C.Serumaliquotsof300 L weretransferredintomicrotubesandstoredat Š 80Cfor lateranalysisofinsulin,leptin,adiponectin,andhs-CRP. Allassayswereperformedinduplicateinasingleassay usingcommerciallyavailableELISAkitsaccordingtothe manufacturer ’ sinstructions(leptinandadiponectin:R&D SystemsInc.,Minneapolis,MN,USA;hs-CRPandinsulin: IBLInternational,Inc.,Hamburg,Germany).Themeanof duplicatesampleswereusedforstatisticalanalysis.Interassaycoefficientsofvariationwere1.3%,6.2%,and5.2%for leptin,adiponectin,andhs-CRP,respectively.Inter-assay dataweremissingforinsulindu etotechnicalerrors.Intraassaycoefficientsofvariationwere5.6%,13.8%,3.8%,and 8.1%forinsulin,leptin,adiponectin,andhs-CRPrespectively.Insulinresistancewasassessedusingthehomeostatic modelofassessment,asdescribedpreviously[19].ComplianceCompliancewascheckedbyaskingparticipantstobring supplementcontainersevery4weeks.Weeklycallsor emailsweresenttoremindparticipantstotakethesupplementandmaintaintheirfoodandphysicalactivity logsforthedurationofthestudy.StatisticalanalysisAn apriori poweranalysiswasperformedwhichrevealeda needforaminimumof6participantspergrouptoachievea powerof0.80, =0.05,standarddeviation=1.1,difference= 6(41).Onewayanalysisofvariance(ANOVA)wasperformedtoexaminepossiblegroupdifferencesatbaseline. Datawereanalyzedusinga2X5repeatedmeasuresANOVA ([PLMIDS])([week0week2week4week6 week8]).DatawereanalyzedusingJMPPRO9software (Cary,NC).Ifsignificantmaineffectswereidentifiedby ANOVA,aTukeyposthoccomparisontestwasperformed tolocatedifferences.Ormsbee etal.JournaloftheInternationalSocietyofSportsNutrition 2014, 11 :37Page3of10 http://www.jissn.com/content/11/1/37

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ResultsParticipantdemographicsAtotalof160individualswerepre-screenedforparticipationinthisstudy.Ofthe68individualseligibleforthe study,34decidedtoparticipate.Outofthese34participants,5participantswithdrewfromthestudyduetopersonalreasons.Atotalof29participantscompletedthe study,however,datafrom7participantswasexcludeddue tolowcompliance(<80%oftotalsupplementintake). Therefore,22participantswith11ineachgroupwereincludedinthestatisticalanalysis.Asidefromfatmass (p<0.05),therewerenostatisticaldifferencesbetween groupsatbaseline(Table1).BodycompositionBodycompositionandanthropometricdataispresented inFigure1andTable2.Therewerenomaintimeor grouptimeinteractionsobservedforthefollowingvariables:bodymass,bodymassindex,percentbodyfat,%androidfat,percentgynoidfat,fatmass,fatfreemass, waistandhipcircumference,andwaisttohipratio. However,asignificanttimeeffectwasobservedforBMI (+1.2and+1.0%)andFM(+1.7and+1.9%)forPLand MIDS,respectively(Table2).Additionally,posthoc analysesrevealednodifferencesbetweengenders.Lipidprofile,heartrate,andbloodpressureThelipidprofilevariablesmeasuredweretotalcholesterol, highdensitylipoprotein,lowde nsitylipoproteinandtriglycerides.Therewasnosignificantmaintimeorgrouptime interactionsobservedforanyvariablesofthelipidprofileor fastingbloodglucose(Table3).Weobservedamaintime andgrouptimeeffectforheartrate.Specifically,heart rateincreasedinPLandwasunchangedinMIDS(PL, + 4.0bpmvs.MIDS, 0bpm,p=0.005).Therewasalsoa maintimeandgrouptimeeffectfordiastolicbloodpressure(PL, +2mmHgvs.MIDS, -1mmHg,p=0.04) withnochangesinsystolicbloodpressure(Figure2).FoodintakeDietaryintakeoftotalcalories,carbohydrates,proteins, andfatswasnotdifferentatbaselineanddidnotchange Figure1 Free-fatmass,fatmass,andpercentbodyfatchanges pre-topost-intervention.A ,freefatmasspre-andpost-intervention. B ,fatmasspre-andpost-intervention. C ,bodyfatpercentpre-and post-intervention.FFM,free-fatmass;kg,kilogram;PL,placebo;MIDS, multi-ingredientdietarysupplement;FM,fatmass;BF,bodyfat;%, percent.*denotesthatgroupsweresignificantly(P<0.05)differentat week0(pre). Table1Participantdemographicsatweek0(N=22)VariablesPL[n=11 (7women,4men)] MIDS[n=11 (9women,2men)] p Age(years) 33.83.535.93.40.67 Height(cm) 169.23.3173.22.90.36 BM(Kg) 96.96.891.95.60.57 %BF 43.62.540.52.40.45DataaremeanSEM.PL,Placebo;MIDS,multi-ingredientdietarysupplement; BM,bodymass;BMI,bodymassindex;%BF,percentbodyfat;FM,fatmass; FFM,fatfreemass;W/Hratio,waittohipratio.Ormsbee etal.JournaloftheInternationalSocietyofSportsNutrition 2014, 11 :37Page4of10 http://www.jissn.com/content/11/1/37

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after8weeksofsupplementationregardlessofgroup. DataarepresentedinTable4.HungerscaleRatingsofhunger,satiety,anddesiretoeatarepresented inFigure3.Nosignificantgrouptimeinteractionsor timeeffectswereobservedforsatietyordesiretoeat. However,asignificantgrouptimeinteractionwas observedforhunger(PL, -15.8mmvs.MIDS, + 10.3mm,p=0.04).MoodstateNosignificantgrouptimeinteractionsortimeeffects wereobservedforanyofthemoodstatevariables.However,amaintimeeffect(p=0.02)wasobservedfortension Table2Bodycompositionandanthropometricsatweek0andweek8VariablesGroupnWeek0Week8 Time p Timegroup p BM(kg) PL1196.96.897.97.1+1.00.240.70 MIDS1191.95.692.45.9+0.5 BMI(kg/m2) PL1134.11.834.51.9+0.40.020.62 MIDS1130.01.530.31.5+0.3 %BF PL1143.62.543.82.4+0.20.380.91 MIDS1140.52.440.72.6+0.2 %Androidfat PL1149.93.249.22.8 0.70.850.07 MIDS1144.82.845.53.1+0.7 %Gynoidfat PL1146.52.946.72.7+0.20.630.23 MIDS1143.02.642.62.8 0.5 FM(kg) PL1141.94.842.64.9+0.70.040.84 MIDS1136.24.036.94.3+0.7 FFM(kg) PL1154.73.254.93.0+0.20.240.50 MIDS1154.73.355.23.3+0.5 Waist(cm) PL11107.04.3105.23.9 1.80.320.69 MIDS1199.93.299.13.3 0.8 Hip(cm) PL11121.04.9120.64.7 0.50.590.90 MIDS11114.03.3113.73.6 0.2 Waisttohipratio PL110.890.020.880.02 0.010.480.73 MIDS110.880.020.870.03 0.01DataaremeanSEM;PL,Placebo;MIDS,multi-ingredientdietarysupplement;BM,bodymass;BMI,bodymassindex;%BF,percentbodyfat;%Androidfat, percentAndroidfat;%Gynoidfat,percentgynoidfat;waist,waistcircumference;hip,hipcircumference. changefromweek0toweek8. Table3Lipidprofileatweek0andweek8VariablesGroupnWeek0Week8 Time p Timegroup p TC(mg/dl) PL11179.77.9183.88.8+4.00.390.91 MIDS11178.98.6184.214.2+5.0 TRG(mg/dl) PL10119.518.7129.723.4+9.00.990.19 MIDS11107.019.396.921.1 10 HDL(mg/dl) PL1156.15.150.65.1 5.00.390.27 MIDS1152.43.653.13.9+1.0 LDL(mg/dl) PL10102.110.7109.37.4+7.00.120.96 MIDS11105.18.6111.812.1+7.0 LDL/HDL PL102.20.42.40.3+0.20.140.68 MIDS112.10.32.30.4+0.1DataaremeanSEM;PL,Placebo;MIDS,multi-ingredientdietarysupplement;TC,totalcholesterol;TRG,triglycerides;HDL,highdensitylipoprot ein;LDL,low densitylipoprotein;HR,heartrate;SBP,systolicbloodpressure;DBP,diastolicbloodpressure;nisdifferentforTRG,LDL,LDL/HDLasaresultofi nsufficientdata. changefromweek0toweek8.Ormsbee etal.JournaloftheInternationalSocietyofSportsNutrition 2014, 11 :37Page5of10 http://www.jissn.com/content/11/1/37

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(PL, -3.78vs.MIDS, – 2.2)andconfusion(PL, -3.2; vs.MIDS, -1.2).HormonesFastingconcentrationsofinsulin,leptin,adiponectin, andhs-CRParepresentedinTable5.Nosignificanttime effectsorgroupbytimeinteractionswereobservedfor anyofthemeasuredhormones.Additionally,insulinresistancewasnotdifferentbetweengroupsovertimeand nosignificantchangeswereobservedineithergroup after8weeksoftreatment. Nosignificantgroupbytime interactionsortimeeffectswereobservedforadiponectintoleptin(A/L)ratioforeithergroup(Table5).However,A/Lratios weresignificantlyhigherinMIDSatbaselineandpostinterventioncomparedtoPL.ReportedsideeffectsInPL,therewasareportofinsomnia(n=1)andbloating (n=1).SideeffectsreportedforMIDSwereacne(n=1), jitteriness(n=1),nausea(n=1),weightgain(n=1),and lossofappetite(n=1).Noothersideeffectswerereported ineithergroup.DiscussionTheprimaryfindingsofthisstudywerethat8weeksof MIDSsupplementationinoverweight/obesemenand womendidnotreducebodymassorimprovebodycompositionversusPL.Furthermore,endocrinemarkers(insulin,leptin,adiponectin,andhs-CRP),bloodlipids,satiety, foodintake,andmoodstatevariableswerenotsignificantlydifferentorafter8weeksregardlessofgroup. Theresultsofthepresentstudyareinoppositiontorecentreportsofenhancedbodycomposition,moodstate, andmentalfocuswiththeconsumptionofothermultiingredientdietarysupplements[6,7].Inthesestudies, supplementswereconsumedwhichcontainedgreentea extractsandcaffeineamongotheringredients.AsMIDS sharedtheseingredientsincommonwiththesupplements usedinthesestudies,thelackofimprovementinbody compositionwasunexpected. Figure2 Systolicanddiastolicbloodpressureovertime. BP,bloodpressure;mmHg,miligramsofmercury;PL,placebo;MIDS, multi-ingredientdietarysupplement.*denotesthatPLatweek8was significantly(p<0.05)differentthanMIDSatweeks0,2,4,and8. Table4Foodintakeatweek0andweek8VariablesGroupnWeek0Week8 Time p Timegroup p Calories(kcal) PL102666.2283.12366.9343.1 299.30.360.41 MIDS112846.0244.62478.5377.8 367.5 Carbohydrate(g) PL10335.237.1271.3539.8 63.850.270.63 MIDS11333.537.3299.055.1 34.53 Carbohydrate(%) PL1050.92.846.33.3 4.60.310.54 MIDS1146.93.247.92.5+1.0 Protein(g) PL1093.010.697.216.2+4.20.270.60 MIDS11112.310.095.412.1 16.9 Protein(%) PL1014.81.716.61.4+1.80.880.55 MIDS1116.82.116.31.5 0.5 Fat(g) PL10101.914.791.918.0 100.550.06 MIDS11119.915.8101.616.6 18.3 Fat(%) PL1033.12.134.43.2+1.80.130.08 MIDS1137.12.536.62.3 0.5DataaremeanSEM;PL,Placebo;MIDS,multi-ingredientdietarysupplement. changefromweek0toweek8.Ormsbee etal.JournaloftheInternationalSocietyofSportsNutrition 2014, 11 :37Page6of10 http://www.jissn.com/content/11/1/37

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Indeed,researchexamininggreenteaandcaffeine eitheraloneorincombinationhaveconsistently reportedtheseingredientstoreducebodymassandfat massinoverweightorobeseindividuals[8,20-22].Moreover,themechanismsfortheseeffectsarealsowellresearched.CaffeinestimulatesthereleaseofcatecholaminesandinhibitsdegradationofcAMP,which enhancessympatheticstimulation[11,23].Similarly, greenteacanalsoincreasethermogenesisasitcontains caffeineandnumerouspolyphenols(catechins,epicatechin,epigallocatechin,andtheirgallates).Epigallo CatechinGallate(EGCG),themostabundantpolyphenol inGT,preventscatecholaminedegradationviainhibitionofcatechol-o-methlytransferase(COMT)enzyme activity[10].This,similartotheeffectsofcaffeine,can effectivelyprolongnorepinephrine-inducedsympathetic stimulationtherebyenhancingthermogenesis[10]. Thelackofsignificantchangeinbodymassorcompositioncanbeexplainedbyanumberoffactors.Interestingly theimpactofcaffeinemaybeattenuatedbythedevelopmentoftolerance.Robertsonetal.[24]demonstratedthat caffeine(250mg/day)increasedplasmacatecholamines after3daysofingestioninhealthymenandwomen.However,after14daysofcontinuoussupplementationthe plasmacatecholamineconcentrationsdecreasedandwere comparabletotheplacebo[24].Theparticipantsinour studyweremoderateconsumersofcaffeine(~137mg/day asself-reported;range:0to~618mg/day),possiblyresultinginattenuatedsensitivitytocaffeinesupplementation duringthetrialperiod.Thelackofchangeinheartrate andbloodpressureinMIDSappearstoconfirmthis.The ineffectivenessofcaffeineinthecurrentstudymayalsobe relatedtogeneticfactors.Indeed,Womacketal.[25] notedageneticpolymorphismwhichinfluencesone ’ s Figure3 Hunger,satiety,anddesiretoeatratingspre-andpost-intervention.A ,hungerratingspre-andpost-intervention. B ,satiety ratingspre-andpost-intervention. C ,desiretoeatratingspre-andpost-intervention.mm,milimeters;PL,placebo;MIDS,multi-ingredientdietary supplement.*denotesagrouptimeinteraction(p<0.05). Table5Endocrinechangesatweek0andweek8VariablesGroupnPrePost Time p Timegroup p Insulin( IU/mL) PL1118.83.617.21.5 1.60.570.16 MIDS1011.41.215.12.0+3.7 Leptin(pg/mL) PL1152.211.442.08.1 10.20.130.58 MIDS1130.77.625.97.5 4.8 Adiponectin( g/mL) PL115.90.86.51.2+0.60.730.12 MIDS119.81.78.91.5 0.9 hs – CRP(mg/L) PL114.41.14.61.3+0.20.890.37 MIDS102.80.92.50.6 0.3 Glucose(mg/dl) PL1199.03.398.24.4-0.80.940.78 MIDS1196.34.396.95.0+0.6 HOMA-IR PL114.660.984.190.42 0.470.730.19 MIDS102.820.443.610.52+0.81 A/L PL110.360.10.440.1+0.080.130.39 MIDS111.10.21.30.23+0.2DataaremeanSEM;PL,Placebo;MIDS,multi-ingredientdietarysupplement;hs-CRP,highsensitivityCreactiveprotein;HOMA-IR,homeostaticmod elassessment insulinresistance;A,Adiponectin;L,Leptin; n isdifferentforinsulinandhs-CRPasdatawasmissingduetotechnicalerrors. changefromweek0toweek8.Ormsbee etal.JournaloftheInternationalSocietyofSportsNutrition 2014, 11 :37Page7of10 http://www.jissn.com/content/11/1/37

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responsivenesstocaffeineintermsofexerciseperformance.Perhapsthemajorityofsubjectsinourstudywere ‘ non-responders ’ tocaffeinealthoughthisispurelyspeculation.Additionally,ameta-analysisonthebodymassand compositioneffectsofcaffeineandGTnotedethnicity tobeasignificantfactor[26].Indeed,caffeineandGT mayhavemorepronouncedeffectsonAsiansversus Caucasians.Thismaybeduetogeneticdifferencesin COMTactivity[27].Asourstudywascomposedofvariousethnicities(e.g.Caucasian,AfricanAmerican,and Asian),thismayhaveincreasedvariabilityresultinginthe observednon-significanteffects.Finally,thecomposition ofthemulti-ingredientdietarysupplementsutilizedin otherstudies[6,7]issignificantlydifferentfromMIDS. Thus,theinfluenceofotheringredientsmayexplainthe benefitsreportedinthesestudies.Anotherpossibilityis thattheingredientsintheseothersupplementsmaysynergisticallyenhancebodycomposition.Alternatively,the potentialbenefitsofthegreenteaandcaffeineinMIDS mayhavebeencounteractedbysomeofitsotheringredients,buttheexperimentaldesigndidnotallowforexaminationoftheweighted-effectsofthevariousingredients. Ourfindingsofalackofchangeinbodymassorcompositionisalsosomewhatunexpectedbasedonreported benefitsofCLAingestion.CLA,afattyacidabundantin seedoils,mayaidintheregulationoflipidmetabolism. Specifically,CLAhasbeenreportedtoreduceuptakeof lipidsintoadipocytesbyinhibitinggeneexpressionand activityofstearoyl-CoAdesaturaseandlipoproteinlipase [28,29].Additionally,CLAseemstoincreasetheactivity ofCPT1therebyincreasingfattyacidoxidation.These mechanismsmayexplainimprovementsinbodycompositionthathavebeenconsistentlyreportedwithCLA supplementationinanimals[28,30]. ThereissomeevidencetosuggestthatCLAimproves bodycompositioninhumans.Blanksonetal.[31],studied overweight/obeseparticip antsthatconsumedvarious amountsofCLA(1.7,3.4,5.1,6.8g/day)followingexercise for12wks.Asaresultoftheintervention,participantswho receivedthehighestdoseofCLA(6.8g/day)increasedlean masstoagreaterdegreethantheothergroups.Inaddition, decreasesinfatmasswerereportedinthegroupsreceiving 3.4and6.8g/dayCLA,(butnot5.1g/day).Othershave alsoreportedenhancedleanmassregainfollowingweight lossinoverweightparticipantswith1.8to3.4g/dayofCLA [32]andreduced%BFinhealt hy,normalweight,individualswith4.2g/dayofCLA[33].Withthisinmind,our findingsofnochangeinbodymassorcompositioncould betheresultofsub-optimaldosesofCLA.Inthecurrent study,theproprietaryblendconsumedinMIDScombined thedoseofCLAwithBCAA(2.52g/daycomposedofCLA andBCAAs).Thus,evenifthemajorityoftheproprietary blendwascomposedofCLA,thedosewouldstilllikelyfall atthelow-endofreportedeffect ivedosages(1.8-6.8g/day). Alternatively,itisalsopossiblethatthebenefitsofCLAare onlyrealizedwhencombinedwithexercise[31].Participantsinourstudywereinactivepriortoandduringthe study,whichmayhaveinfluencedtheeffectivenessofCLA. Whateverthecasemaybe,ourfindingsareinlinewithnumerousothersreportingthatCLAsupplementationhasno effectonbodymassorcomposition[34-36].Furtherstudy onthetrueeffectsofCLAonbodymassandcomposition inhumansinclearlywarranted. Inthepresentstudy,bodycompositionwasnotimproved despitetheBCAAcontentofMIDS.Increasingdietary proteinhelpstoreduceweight,andpreserveleanbody massinhealthyadults[37],andsomeofthebenefitsmay beattributabletoBCAA[38].Theseeffectsmay beafunctionofthestimulatoryeffectthatcertainBCAA (e.g.leucine)haveonmuscleproteinsynthesis[17].Positive effectsfromBCAAconsumptiononbodycompositionare oftenreportedwhencombinedwithexercise[39].However, theinfluenceofBCAAonbodycompositionininactiveindividualsisrelativelyunknown.Ourfindingssuggestthat consumingadditionalBCAA(< 2.52g/day)incombination withtheotheringredientsinMIDShasnoeffectonbody composition.Alternativel y,perhapsahigherdoseis requiredtorealizepotentialbenefits.Indeed,manyanimal studiesreportingBCAA-indu cedlossesinfatmassutilize >5g/day[40]. MaintaininglowlevelsofLDL,TC,andTRGandhigh levelsofHDLmayhelptopreventcardiovasculardisease [41].OurfindingsdonotsupportMIDSasbeingcardioprotectiveasweobservednochangesinbloodlipids.Some studieshavereportedcaffeine,greentea,and/orCLAtoreduceLDL,TC,andTRGwhilemaintainingorincreasing HDL[22,31,42].However,th edosesusedinthesestudies weretypicallyhigherthaninth ecurrentstudyand/orwere combinedwithexerciseordiet aryinterventions.Perhaps morerobustdietaryand/orphy sicalactivitychangesarerequiredtoseesignificantchangesinbloodlipids. EndocrinebiomarkersweresimilarlyunaffectedbyMIDS orPL.Thiswasnotaltogetherunexpectedastheingredients inMIDSmayhaveopposingeffectsonthehormones measured.Forinstance,caffeineandCLAareknowntoreduceinsulinsensitivity[43,44] .However,caffeineandgreen teahavealsobeenreportedtoincreaseadiponectin,ahormoneproducedinadiposethatmaypreventinsulinresistance[45,46].Theseopposingeffectsmayexplainwhyinsulin andfastingglucosewereunchangedwithMIDS.Leptin,a hormonethatmayregulateenergyintake,andhs-CRP,a proinflammatorycytokineimplicatedinatherogenesis,were similarlyunchangedfollowing MIDSsupplementation.AlthoughthereissomeevidencesuggestingthatCLAmayreduceleptinlevels[47],theeffectsoftheotheringredientson leptinandhs-CRParerelative lyunknown.Theresultsofthe currentstudysuggestthatMIDSconsumptiondoesnotalter leptin,hs-CRP,orhormonesrelatedtoinsulinsensitivity.Ormsbee etal.JournaloftheInternationalSocietyofSportsNutrition 2014, 11 :37Page8of10 http://www.jissn.com/content/11/1/37

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SatietywasalsounaffectedbyMIDSsupplementation. Whenattemptingtoloseweight,increasingsatietymay helptoreduceenergyintake.Previousstudieshavereportedenhancedsatietywithincreaseddietaryprotein intake[37].Additionally,caffeinesupplementationmay reducespontaneousenergyi ntake[48].Nevertheless, weobservednochangedinsatietyorfoodintake.As mentioned,oursubjectsweremoderatecaffeineconsumers,whichlikelyattenuatedanycaffeine-mediated effectsonsatiety.Moreover,participantsinthecurrent studydidnotincreasedietaryproteinintake,butrather supplementedwitharelativelysmalldoseofBCAA. PerhapsBCAAs-alonemayhaveminimaleffectsonsatiety,orthedosewastoosmalltoelicitanychangesin satiety.Itisworthnotingthatparticipantsconsuming MIDSreportedanincreaseinhunger.Astudyby Chiouetal.[49]reportedthatparticipantstakinga purporteddietarysupple ment,whichwasactuallya non-caloricplacebo,showedapreferenceforabuffet mealratherthananorganicmeal.Furthermore,subjectsreceivingtheshamsupplementalsoshowedreduceddesiretoexercise[49].Inthecurrentstudy,itis possiblethatparticipants thatbelievedtheywerereceivingadietarysupplementfeltincreasedlicensefor hedonicbehaviorpossiblycon tributingtotheincreased hungerratings.However,itshouldbenotedthatinthe presentstudyparticipants ineithergroupdidnotincreasefoodintake.Thus,furtherstudyonthetrue effectsofMIDSonsatietyandhungerarewarranted. Inconclusion,consumptionofMIDSfor8wksinoverweightandobese,butotherwisehealthy,menandwomen resultedinnochangesinbodymassorcomposition, bloodlipids,endocrinebiomarkers,moodstate,orsatiety. Infuturestudies,largerdosesofMIDSshouldbeadministered,andtheeffectsofMIDSincombinationwithdiet and/orexerciseshouldbeinvestigated.Abbreviations MIDS: Multi-ingredientdietarysupplement;PL:Placebo;GT:Greentea; CLA:Conjugatedlinoleicacid;BCAA:Branchedchainaminoacids. Competinginterests Theauthorsdeclarethattheyhavenocompetinginterests. Authors ’ contributions MJOdesignedandmanagedthestudy,securedfunding,analyzedthedata anddraftedthemanuscript.SRRcarriedoutallpracticalaspectsofthestudy andassistedwithdataanalysis,andmanuscriptpreparation.MTSassisted withstudydesignandmanuscriptpreparation.DAB,AWK,MEE,NF,TAM, andDTassisteddatacollectionandanalysisandmanuscriptpreparation. Allauthorsreadandapprovedthefinalmanuscript. Acknowledgements ThisstudywassupportedbyagrantfromtheInternationalSocietyof SportsNutritiontoMJO.WewouldliketothankEmeryWardandWyatt Eddyfortheirhelpwithdatacollection.Wewouldalsoliketothankallof thevolunteersforthisstudy. Authordetails1DepartmentofNutrition,FoodandExerciseSciences,FloridaState University,Tallahassee,FL32306,USA.2UniversityofKwaZulu-Natal,Durban, SouthAfrica.3InstituteofSportsScience&Medicine,FloridaStateUniversity, Tallahassee,FL,USA. Received:23June2014Accepted:18July2014 Published:26July2014 References1.OgdenCL,CarrollMD,KitBK,FlegalKM: Prevalenceofobesityinthe UnitedStates,2009 – 2010. NCHSDataBrief 2012, 82: 1 – 8. 2.MustA,SpadanoJ,CoakleyEH,FieldAE,ColditzG,DietzWH: Thedisease burdenassociatedwithoverweightandobesity. JAMA 1999, 282: 1523 – 1529. 3.BarteJCM,terBogtNCW,BogersRP,TeixeiraPJ,BlissmerB,MoriTA, BemelmansWJE: Maintenanceofweightlossafterlifestyleinterventionsfor overweightandobesity,asystematicreview. ObesRev 2010, 11: 899 – 906. 4.KernanWN,ViscoliCM,BrassLM,BroderickJP,BrottT,FeldmannE, MorgensternLB,WilterdinkJL,HorwitzRI: Phenylpropanolamineandthe riskofhemorrhagicstroke. 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