or Linoleic Acid-Containing Meals on Appetite and Metabolic Markers

17 Pages·2017·2.72 MB·English

nutrients
Article
The Acute Effect of Oleic- or Linoleic
Acid-Containing Meals on Appetite and Metabolic
Markers; A Pilot Study in Overweight or
Obese Individuals
ShaanS.Naughton1,ErikD.Hanson1,†,MichaelL.Mathai1,2andAndrewJ.McAinch1,3,*
1 InstituteforHealthandSport,VictoriaUniversity,POBox14428,Melbourne,Victoria8001,Australia;
[email protected](S.S.N.);[email protected](E.D.H.);
[email protected](M.L.M.)
2 TheFloreyInstituteofNeuroscienceandMentalHealth,TheUniversityofMelbourne,30RoyalParade,
Parkville,Melbourne,VIC3052,Australia
3 AustralianInstituteforMusculoskeletalScience(AIMSS),CollegeofHealthandBiomedicine,
VictoriaUniversity,Melbourne,Victoria8001,Australia
* Correspondence:[email protected];Tel.:[phone]
† PresentAddress:DepartmentofExerciseandSportScience,UniversityofNorthCarolina,315Woollen,
CB#8605,ChapelHill,NC27599,USA.
(cid:1)(cid:2)(cid:3)(cid:1)(cid:4)(cid:5)(cid:6)(cid:7)(cid:8)(cid:1)
(cid:1)(cid:2)(cid:3)(cid:4)(cid:5)(cid:6)(cid:7)
Received:23August2018;Accepted:21September2018;Published:26September2018
Abstract: Despite the abundance of plant-derived fats in our diet, their effects on appetite,
andmetabolicmarkers,remainunclear. Thissingle-blinded3-waycross-overpilotstudyaimedto
investigatetheabilityofthetwomostabundantdietaryplant-derivedfats,oleic(OA)andlinoleic
(LA) acids, to modulate postprandial appetite and levels of circulating appetite and metabolic
regulatorsinoverweight/obeseindividuals. Mealswereahigh-carbohydratecontrol,ahigh-OA
orahigh-LAmeal,andprovided30%ofparticipants’estimatedenergyrequirements. Mealswere
consumedafteranovernightfast,withbloodsamplescollectedover31/4h. Appetiteparameters
wereassessedviaavalidatedvisualanaloguescalequestionnaire. Hormonesandothercirculating
factorswerequantifiedusingmultipleximmunoassays. Eightparticipants(age45.8±3.6(years),
bodymassindex32.0±1.3(kg/m2))completedthestudy. Allmealssignificantlyincreasedfullness
andreduceddesiretoeat. Thecontrolandhigh-OAmealssignificantlydecreasedprospectivefood
intake. The high-LA meal increased ghrelin levels (p < 0.05), a hormone which encourages food
intake. Thiswascoupledwithasignificantacuteincreaseinresistinlevels,whichimpairsinsulin
signaling. Takentogether,thisstudyindicatesthatinoverweight/obeseindividuals,high-LAmeals
maypromoteexcessenergyintakeandalterglucosehandling,thoughalargercohortmayberequired
tostrengthenresults.
Keywords: dietaryfats;appetiteregulation;ghrelin;satiety;oleicacid;linoleicacid
1. Introduction
Appetiteisthemainregulatoroffoodintake,andassuchisakeycomponentintheregulation
ofenergyintakeandbodyweight[1]. Appetiteismodulatedbyarangeofendogenoussubstances
producedbythestomach(ghrelin),smallintestine(cholecystokinin,glucose-dependentinsulinotropic
peptide(GIP)),pancreas(insulin),adiposetissue(adiponectin,resistin)andbrain(endocannabinoids),
with many of these operating together, as well as in opposition [2]. Despite this, it is believed
that when high-fat foods are available, as are found abundantly in current Western cultures,
passive overconsumption occurs [3]. This is believed to be due to the body’s inability to gauge
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Nutrients2018,10,1376 2of17
energyintakeaccurately,potentiallyleadingtochronicexcessesofenergyintake,whichmayleadto
weightgainandobesity[3]. Additionally,thereisthepossibilitythatourhormonalcontrolofappetite,
whichevolvedinconjunctionwithalow-fatdiet,maybedesensitizedtoovernutritionbymodern
high-fatdietarypatterns[4,5]. Withtheexceptionofthefirstmealoftheday,mostmealsareconsumed
by humans during the postprandial state [6]. As such, understanding how postprandial appetite
regulationisinfluencedbyourcurrentnutritionalenvironmentisimportanttounderstandhowour
foodintakeaffectssubsequentintakeand,ultimately,weightstatus.
Whencomparingthesatiatingeffectofthemacronutrients,lipidsaretheleastsatiatingonan
energybasis,possiblyleadingtoexcessiveenergyintake[7]. Despitethis,theabilityofspecificfatsto
modulateappetiteandfoodintakeisyettobefullyelucidated. Thisstudyfocusesonoleicacid(OA),
an-918carbonmonounsaturatedfat,foundprimarilyinoliveoil,thoughalsoinotherplantbasedoils,
nuts,andvegetables,makingitthemostcommondietarymonounsaturatedfat[8],andalsoonlinoleic
acid(LA)an-618carbonfattyacid,themostcommonlyconsumedpolyunsaturatedfat,beingfound
predominatelyinplantoils,nuts,meat,andanimalproducts[9]. Thisisanimportantareaofresearch,
especiallyasthedietaryavailabilityofLAintheUSA[9],Australia[10],Ireland[11],Barbados[12],
Switzerland[13],Europe[14],andChina[15],hasincreaseddramatically(forexample,over2.5-fold
intheUSA[9])inrecenttimes. LAistheprecursorofthetwomainendocannabinoids,anandamide
and2-arachidonylglycerol,bothofwhichpromotehedonicfoodintakeandadiposetissueenergy
storage[16]. Additionally,theendocannabinoidsystemisupregulatedinobesity,whichmayfurther
perpetuateexcessiveenergyintake[17]. TheimportanceofLAinthepostprandialstateisnotonlydue
toitspotentialtomodulatehungerandsatiety,itisalsotheprecursorofarachidonicacid,whichcan
beconvertedtoseveraldifferentinflammatorymediators[18]. Assuch,thereisapossibilitythata
highintakeofLAcouldperpetuatethelowgradesystemicinflammationfoundinoverweightand
obeseindividuals[19].
IncomparisontoLA,OAisnotinvolvedinthesynthesisofinflammatorymediators,withseveral
studiesfindingthatOAanditsendocannabinoidderivativeoleoylethanolamide(OEA),reducehunger
andsubsequentfoodintake[8,20,21]. Despitethis,studiescomparingOA-andLA-containingmeals
in an acute time frame have failed to find a difference in perception of hunger or appetite in the
postprandialperiod[22,23].
The influence of OA or LA on blood glucose regulation and the development of diabetes
is controversial and currently unclear [24–27]. Additionally, the role of these specific fatty acids
(FA)inacutelymodulatingsatiety-regulatinghormonesandperceptionofhungerandprospective
food intake is yet to be fully elucidated, with no clear associations identified between them and
appetite-modulatinghormones[28]. Therefore,thispilotstudyaimedtoassesstheacute(2h)response
ofappetite-regulatinghormones,metabolicmarkers,andcytokines,aswellasself-reportedappetite
parameters,followingtheconsumptionofmixedcompositionmealscontaininghighlevelsoftwo
ofthemostcommondietaryfats(LAorOA),incomparisontoanenergy-matchedpredominately
carbohydrate-containingmeal,inoverweightorobesemenandwomen.
2. MaterialsandMethods
2.1. StudyDesign
This study was designed as a single-blinded, 3-way cross-over pilot study. Due to the cost
associated with measuring the circulating factors, many of which have not been investigated in
response to acute FA intake, this pilot was undertaken to determine the efficacy of the protocol.
With an aim of 10 subjects completing the study, the study had >80% power to discriminate
a 1 standard deviation difference in plasma ghrelin levels between meals at significance level of
p<0.05. EthicalapprovalwasobtainedfromtheVictoriaUniversityhumanresearchethicscommittee
(HRETH 12/87), complying with the principles of the declaration of Helsinki. Participants were
recruitedMay–August2012throughflyersdistributedoncampusandviaglobalemail,withinformed
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consentobtainedfromparticipants. ThestudytookplaceintheUniversity’steachingclinicaspart
oNfuatriyenetas r2-0l1o8n, 1g0,r xe FsOeaRr PchEEpR rRoEjeVcIEt.WE ach participant consumed the test meals after an overnight3f aofs 1t,7
withonemealconsumedeachweekinarandomorder,withaminimum5dayperiodbetweenmeals.
consumed each week in a random order, with a minimum 5 day period between meals. Inclusion
Inclusioncriteriaforthestudywasabodymassindex(BMI)>25kg/m2andawaistcircumference
criteria for the study was a body mass index (BMI) >25 kg/m2 and a waist circumference >94 cm
>94cm(male)/>80cm(female),weightstable(noself-reportedintentionalorunintentionalweight
(male)/ >80 cm (female), weight stable (no self-reported intentional or unintentional weight loss or
loss or gain over the preceding three months), and aged 18–60 years. Exclusion criteria were the
gain over the preceding three months), and aged 18–60 years. Exclusion criteria were the self-reported
self-reported presence of Type I or II diabetes mellitus, insulin resistance, heart, liver or kidney
presence of Type I or II diabetes mellitus, insulin resistance, heart, liver or kidney disease, pregnancy
disease, pregnancy (or planning to become pregnant), breastfeeding, and the use of weight loss
(or planning to become pregnant), breastfeeding, and the use of weight loss
medication/supplementationorFAsupplementation. Forfurtherdetailsrelatingtothestudydesign
medication/supplementation or FA supplementation. For further details relating to the study design
pleaseseetheCONSORTchecklistinTableS1. Bodycomposition(fatmass%andleanmass%)of
please see the CONSORT checklist in Table S1. Body composition (fat mass % and lean mass %) of
fastedparticipantswasmeasuredusingdual-energyx-rayabsorptiometry(DXA)(HologicDiscovery
fasted participants was measured using dual-energy x-ray absorptiometry (DXA) (Hologic Discovery
QDR, Hologic (Australia) Pty Ltd., North Ryde, NSW, Australia) in the first week of the study.
QDR, Hologic (Australia) Pty Ltd., North Ryde, NSW, Australia) in the first week of the study.
Anthropometricmeasurementsweretakentwiceduringthestudyandincludedheightmeasurement,
Anthropometric measurements were taken twice during the study and included height
weight (Tanita HD-351 Digital Weight Scale, Tanita Pty. Ltd., Tokyo, Japan), waist circumference
measurement, weight (Tanita HD-351 Digital Weight Scale, Tanita Pty. Ltd., Tokyo, Japan), waist
(measuredatthenarrowestpointbetweenthelowercostalborderandtheiliaccrest),hipcircumference
circumference (measured at the narrowest point between the lower costal border and the iliac crest),
(takenatthelevelofthegreatestposteriorprotuberanceofthebuttocks),bloodpressureandheartrate
hip circumference (taken at the level of the greatest posterior protuberance of the buttocks), blood
(Omron5Series,OmronHealthcare,Inc.,Palatine,IL,USA).
pressure and heart rate (Omron 5 Series, Omron Healthcare, Inc., Palatine, IL, USA).
Meal challenge sessions took place according to the timeline shown in Figure 1. Participants
Meal challenge sessions took place according to the timeline shown in Figure 1. Participants
wereinstructedtoconsumeanutritionallybalancedmealthenightbeforetestingandrefrainfrom
were instructed to consume a nutritionally balanced meal the night before testing and refrain from
alcoholintake. Uponarrival,participantshadacannulainsertedintotheirantecubitalvein. Allmeals
alcohol intake. Upon arrival, participants had a cannula inserted into their antecubital vein. All meals
wereconsumedatthesametimeofday(between9a.m. and10a.m.) tocontrolfordiurnalhormonal
were consumed at the same time of day (between 9 a.m. and 10 a.m.) to control for diurnal hormonal
variation, with a maximum of 15 min provided for consumption. Participants were encouraged
variation, with a maximum of 15 min provided for consumption. Participants were encouraged to
toconsumewaterduringthesessionstomaintainfluidlevelsandassistinthecollectionofblood,
consume water during the sessions to maintain fluid levels and assist in the collection of blood, and
andinstructedtorefrainfromexerciseonthemorningoftestdays.
instructed to refrain from exercise on the morning of test days.

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2.2. TestMealCompositions
2.2. Test Meal Compositions
Thetestmealswerecomprisedoftoastedbread,reducedsugarjam,icingsugar,anddiffering
The test meals were comprised of toasted bread, reduced sugar jam, icing sugar, and differing
oilcompositions(extravirginoliveoil,saffloweroil,andcoconutoil),resultingincontrol,high-OA
oil compositions (extra virgin olive oil, safflower oil, and coconut oil), resulting in control, high-OA
(high-monounsaturatedfattyacid(MUFA)),andhigh-LA(highpolyunsaturatedfattyacid(PUFA))
(high-monounsaturated fatty acid (MUFA)), and high-LA (high polyunsaturated fatty acid (PUFA))
meals,withallingredientsusedcommonlyfoundinWesterndiets. Themealcompositionstookinto
meals, with all ingredients used commonly found in Western diets. The meal compositions took into
account palatability and the ability of participants to be blinded, with prior testing indicating that
these criteria were met. This meal composition was chosen so as to replicate a prepared mixed
composition meal that people would normally consume, as opposed to a liquid formula. As such,
fiber-enriched bread was used, which also had a higher protein content than conventional white
Nutrients2018,10,1376 4of17
accountpalatabilityandtheabilityofparticipantstobeblinded,withpriortestingindicatingthatthese
criteriaweremet. Thismealcompositionwaschosensoastoreplicateapreparedmixedcomposition
mealthatpeoplewouldnormallyconsume,asopposedtoaliquidformula. Assuch,fiber-enriched
breadwasused,whichalsohadahigherproteincontentthanconventionalwhitebreads. Icingsugar
wasusedtoincreasetheenergydensityofthecontrolmealtomatchthoseoftheelevated-fatmeals
duetoithomogenizingeasilywiththejamandalsoduetoitnotaddinganyfurthernutrientstothe
mealwhichwouldbeabsentintheelevated-fatmeals. Thehigh-fatmealshadcomparableratiosof
allfatsotherthantheonewhichhadbeenelevated, withtheamountofenergyfromtheelevated
fats in each high-fat meal also being matched as shown in Table 1. The two high-fat meals were
isoenergeticandhadmatchedcontributionsfrommacronutrientstoenergycontent. Eachtestmeal
contained13.74±0.658kJ/gasdeterminedbynutritionalanalysis(performedusingXyrisFoodWorks
(XyrisPtyLtd., HighgateHill, QLD,Australia)). Themealswereweighedtothenearest0.1gram
and provided 30% of estimated daily energy intake, a figure based on the findings of large-scale
studiesindicatingthatbreakfastsgenerallyaccountfor20–25%ofdailyenergyintake[30,31]. Thiswas
increasedby5%toaccountforthesessionsrunningthroughtolatemorning,removingenergyintake
fromfoodsgenerallyconsumedduringthistimeperiod. Estimatedenergyintakewascalculatedusing
theMifflin–St. Jeorequationwhichhasbeenfoundtobethemostaccurateforoverweightandobese
individuals[32,33]. Thisequationtookintoaccountage,sex,height,andweightandwasadjusted
usingappropriateactivityfactorsbasedonparticipant-reportedusualactivitylevels[34]. Hotwater
andlemonwasprovidedwiththemealstoincreaseparticipantcomplianceandpalatability,caffeinated
beverageswerenotallowedwhilefastingorduringthemealsessionsduetocaffeine’sinfluenceon
bloodglucoselevels[35].
Table1.Nutritionalcompositionofexperimentalmealsper100g.
Nutrient Control High-OA High-LA
Carbohydrate(g) 50.9 33.7 33.7
Carbohydrate(%E) 73.46 39.74 39.74
Protein(g) 4.8 5.8 5.8
Protein(%E) 6.91 6.84 6.84
Fat(g) 7.3 21.5 21.5
Fat(%E) 23.12 55.13 55.13
Saturated(g) 2.5 3.1 3.1
MUFA(g) 2.5 15.1 3.1
OleicAcid(g) 2.4 14.8 2.9
PUFA(g) 2.3 3.2 15.2
LinoleicAcid(g) 2.2 3.0 14.9
Fiber(g) 3.2 4.0 4.0
Omega6:Omega3 16.7 13.1 77.2
OA; Oleic acid, LA; Linoleic acid, % E; percentage of total energy; MUFA; monosaturated fatty acid, PUFA;
polyunsaturatedfattyacid.
2.3. BloodCollectionandAnalysis
A total of five blood samples were collected over each 31/4 h meal session, as shown in
Figure 1. Blood glucose (average of triplicates) concentrations (all time points) were measured in
serumsamplesusinganauto-calibratingautomatedsampler(YSI2300StatPlusGlucoseL-Lactate
analyzer, YSI Inc. Life Sciences, Yellow Springs, OH, USA). Levels of satiety and diabetes-related
markersandcytokines(baseline,1hpostmeal,and2hpostmeal)werequantifiedusingmultiplex
immunoassays (Bio-Plex Pro human diabetes 10-plex immunoassay (# 171A7001M) (measuring
C-peptide,ghrelin,GIP,glucagon-likepeptide1(GLP-1),glucagon,insulin,leptin,totalplasminogen
activatorinhibitor-1(PAI-1), resistin, andvisfatin), Bio-PlexPrecisionProhumancytokine10-plex
immunoassay(#171A1001P)(measuringIL-1β,IL-2,IL-4,IL-5,IL-6,IL-10,IL-12(p70),IL-13,interferon
gamma (IFN-γ) and tumor necrosis factor alpha (TNF-α)) and Bio-Plex Pro human adiponectin
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single-pleximmunoassay(#171A7003M))andaBio-Plex© 200multiplexsuspensionarraysystem
(Bio-RadLaboratories,Inc.,Hercules,CA,USA),accordingtothemanufacturer’sinstructions.
2.4. AppetiteAssessment
Participantscompletedanappetitequestionnaireimmediatelybeforeconsumptionofthemeal
andagainafter2h. Theappetitequestionnaireutilizedavisualanaloguescale(VAS)whichhasbeen
found to be a reliable and valid tool for appetite assessment [36]. This comprised eight questions
eachfollowedby100mmhorizontallines,where0mmrepresented“sensationnotfeltatall”and
100 mm represented “sensation felt the greatest” with subjects asked to mark the line at the point
whichcorrespondedtohowtheywerefeelingatthatparticulartime. Thequestionnairedevelopedby
Parkeretal.[29]hasbeenusedinsimilarstudiesassessingtheeffectofmealcompositiononappetite
andgastrointestinalhormonesinhumans[37–39]. Thedistancefromthebeginningofthelinetothe
participants’markwasmeasuredfromtheleft-handsidetothenearest0.5mm.
2.5. StatisticalAnalysis
AllstatisticalanalysiswasperformedusingPrismGraphPadversion7(GraphPadSoftwareInc.,
LaJolla,CA,USA).ForVASscoresa2-wayanalysisofvariance(ANOVA)wasutilized. Difference
betweenbloodglucoselevelsinresponsetotestmealconsumptionateachtimepointwasdetermined
via2-wayANOVAwithTukey’smultiplecomparisonstest,aswaschangebetweentimepointswithin
eachtestmeal. Foranalysisofappetite-regulatingcompoundsandinflammatorymarkers,areaunder
thecurvewasutilized. Thiswascalculatedasthenetchangefrombaselinetoaccountfornegative
peaks,i.e.,reductionsincirculatingconcentrationsovertheperiodofsampling. Analytesthatdidnot
haveafullsetofdatainsidethereportingrangewereexcludedfromanalysis.
3. Results
Eightparticipantscompletedthestudy,5femalesand3males. Forfurtherinformationregarding
participantrecruitmentandnumberspleaseseeFigureS1. Theaverageagewas45.8±3.6yearsand
averageBMI32.0±1.3kg/m2. FurtherparticipantcharacteristicsareshowninTable2.
Table2.Generalparticipantcharacteristics.
Parameter Average±SEM
Age(years) 45.8±3.6
Height(cm) 169.5±2.2
Weight(kg) 92.2±4.5
BMI(kg/m2) 32.0±1.3
Female 102.4±7.1
WaistCircumference(cm)
Male 108.6±7.9
Female 119.6±4.1
HipCircumference(cm)
Male 111.3±5.7
Female 0.83±0.03
Waist-To-HipRatio
Male 0.97±0.01
FatMass(%ofTotalMass) 37.4±2.9
LeanMass(%ofTotalMass) 55.7±2.8
SystolicBloodPressure(mmHg) 128.4±4.1
DiastolicBloodPressure(mmHg) 82.3±3.5
HeartRate(bpm) 73±4.4
FastingSerumGlucose(mmol/L) 4.6±0.1
BMI;bodymassindex,SEM;standarderrorofthemean.
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For the parameters related to appetite, there was a significant reduction in feeling of hunger
(controlp<0.05,high-OAp<0.05,high-LAp<0.05)andasignificantincreaseinfeelingoffullness
(controlp<0.05,high-OAp<0.05,high-LAp<0.05)afterconsumptionofallmealsasshowninFigure2.
Therewasnosignificantchangetoperceivedsatisfactionanddesiretoeatfollowingconsumptionof
allthreeNmutreieantlss 2.0I1n8, r10e,g xa FrOdR tPoEEpRr RoEsVpIeEcWt iv efoodintakeamount(with‘none’beingthebottomof6 tohf e17s cale
and‘Alargeamount’beingthetopofscale),therewasasignificantdecreasefollowingconsumptionof
meals, as shown in Figure 3, thus it is unlikely that these variables influenced perceived hunger as a
thecontrolandhigh-OAmealscontrolp<0.05,high-OAp<0.05),thoughnotfollowingconsumption
result of consumption of the different test meals.
ofthehigh-LAmeal(p=0.183),asshowninFigure2. VASresultsshowednochangestoperception
Blood glucose measurement showed no significant difference between the test meals at any of
offatiguthee, tnimaues peaoi,notsr. aTnhxeireet ywa(tsh ao usigghnitfhiceanht iginhc-rLeaAsem ine ablloeolidc igteludcoasen oconnscigenntirfiatciaonnst tfroellnodwitnog inthcer ease
anxietycpon=su0m.09p5ti)onas ofa arlel steusltt moefatlhs ebectowneseunm bapsteiloinne oafndan 1y h opfotsht meteeasl tcomnesuamls,patisonsh (coowntnroiln p F<i 0g.u05r,e h3ig,hth-usit
isunlikeOlAy tph <a 0t.0th5,e hsiegvh-aLrAia bp l<e 0s.0in5)fl aunedn bceetdwpeeenrc periev-emdeahlu anngde 1r ha psoastr ceosnuslutmofpctioonns (ucomnptrtoiol pn <o 0f.0th5,e hdigihff-erent
OA p < 0.05, high-LA p < 0.05), as shown in Figure 4.
testmeals.

Figure2.Participantvisualanaloguescaleappetiteparameterresponses.(a)Fullness;(b)Satisfaction;
(c)DesirFeigtuoree a2t.; P(adr)tiPcirpoasnpt evcistiuvael afnoaoldogiunet askcael;e( aep)pHetuitne gpearr.ame=tecro rnestrpoolnmsees.a (la,)(cid:4) Fu=llnheisgsh; (-bO) ASa,t(cid:78)isf=achtioignh; -LA.
(c) Desire to eat; (d) Prospective food intake; (e) Hunger. ● = control meal, ■ = high-OA, ▲ = high-
Assessedthroughcompletionofvalidatedvisualanaloguequestionnairesandmeasuredtothenearest
LA. Assessed through completion of validated visual analogue questionnaires and measured to the
0.5mm. Pre=VAS(visualanaloguescalequestionnaire)completed15minpriortoconsumption;
nearest 0.5 mm. Pre = VAS (visual analogue scale questionnaire) completed 15 min prior to
Post=VAScompleted2hpostconsumption.*=p<0.05allmealscomparingpreandpostconsumption.
consumption; Post = VAS completed 2 h post consumption. * = p < 0.05 all meals comparing pre and
†=p<0.05forcontrolmealcomparingpreandpostconsumption.#=p<0.05forhigh-OAcomparing
post consumption. † = p < 0.05 for control meal comparing pre and post consumption. # = p < 0.05 for
preandpostconsumption.Alldatashownasmean±SEM,alldatapointsn=8.
high-OA comparing pre and post consumption. All data shown as mean ± SEM, all data points n = 8.
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Figure 3. Participant visual analogue scale other sensations responses. (a) Anxiety; (b) Nausea;
(c)FatigFuigeu.re =3. cPoanrttircoiplamnet avli,su(cid:4)al= ahnaiglohg-uOeA sc,a(cid:78)le =othhiegrh s-eLnAsa.tiAonsss ersessepdontsherso. u(ag)h Acnoxmietpyl;e (tbio) nNoaufsveaal; id(ca) ted
visualaFnaatliogguue.e ●q u=e csotinotnronla mireeasl,a ■nd =m heigahs-uOreAd, ▲to =th heignhe-aLrAes. tA0s.s5esmsemd .thPrroeu=ghV AcoSm(pvliestuioanl aonf avlaoligduaetesdc ale
questionvnisauiarle a)ncaolomgpulee qteudes1ti5onmnainirepsr aionrd tmoecaosnusreudm top tthioen n;eParoesstt =0.5V mAmS.c Porme =p VleAteSd (v2ishuaplo asntacloognuseu smcaplet ion.
Alldataqsuheostwionnnaasirme)e caonm±pleSteEdM 1,5a mllind aptraiopr otoin ctosnnsu=m8p.tion; Post = VAS completed 2 h post consumption.
All data shown as mean ± SEM, all data points n = 8.
Blood glucose measurement showed no significant difference between the test meals at any
Figure 3. Participant visual analogue scale other sensations responses. (a) Anxiety; (b) Nausea; (c)
of the timFeatipgouein. ●ts .= Tcohnetrroel wmeaasl, a■s =ig hnigihfi-cOaAn,t ▲in =c rheigahs-eLAin. Abslsoesosdedg tlhurocuogshe ccoomnpcleetniotnr aotfi ovanlisdafoteldlo wing the
consumption of all test meals between baseline and 1 h post meal consumption (control p < 0.05,
visual analogue questionnaires and measured to the nearest 0.5 mm. Pre = VAS (visual analogue scale
high-OAqpu<es0ti.o0n5n,ahirieg) hco-mLAplepte<d 105. 0m5i)na pnridorb toe tcwoneseunmpptrieo-nm; Peoastl =a VndAS1 cohmppolesttedco 2n hs puomst pcotinosnum(cpotinontr. olp<0.05,
high-OAApl<l d0a.t0a 5sh,ohwignh a-sL mAeapn <± S0E.0M5, )a,lla dsastah poowinnts inn =F 8i.g ure4.

Figure 4. Blood glucose responses to ingestion of test meals. ● = control meal, ■ = high-OA, ▲ =
high-LA. Measured in plasma using an auto-calibrating automated sampler. * = p < 0.05 all meals
comparing time points. † = p < 0.05 for control and high-LA comparing time points. # = p < 0.05 for
high-LA comparing time points. All data shown as mean ± SEM, all data points n = 8 (average of
triplicates).
Figure 4. Blood glucose responses to ingestion of test meals. = control meal, (cid:4) = high-OA,
(cid:78)= hFiigghu-rLe A4.. BMlooeda sgulurecdosein repsplaosnmseas tuos iinnggesatniona uotfo t-ecsat lmiberaaltsi. n●g =a ucotonmtroalt emdeasla, m■ p=l ehri.gh*-O=Ap, ▲< 0=. 05 all
high-LA. Measured in plasma using an auto-calibrating automated sampler. * = p < 0.05 all meals
mealscomparingtimepoints.†=p<0.05forcontrolandhigh-LAcomparingtimepoints.#=p<0.05
comparing time points. † = p < 0.05 for control and high-LA comparing time points. # = p < 0.05 for
forhigh-LAcomparingtimepoints. Alldatashownasmean±SEM,alldatapointsn=8(average
high-LA comparing time points. All data shown as mean ± SEM, all data points n = 8 (average of
oftriplicates).
triplicates).
Nutrients2018,10,1376 8of17
Nutrients 2018, 10, x FOR PEER REVIEW 8 of 17
WWhheennc coommppaarrininggb beetwtweeeennt itmimeep pooinintstsf oforrt htheeh higighh-L-LAAm meeaal,l,t htheeb bloloooddg gluluccoosseec coonncceenntrtraatitoionna att
ththee1 1h ht itmimeep pooininttw waasss sigignnifiifcicaanntltylyi ninccrreeaasseedd( p(p< < 00..0055)) ccoommppaarreedd ttoo iimmmmeeddiiaatteellyy ppoosstt ccoonnssuummppttiioonn;;
hhoowweevveerr,,n noonneeo offt htheeo oththeerrm meeaalslsw weerreei ninccrereaasseeddf rforommi mimmmeeddiaiatetelylyp poossttc coonnssuummppttiioonn ttoo 11h hp poosstt
ccoonnssuummpptitoionn..T Thhereerew wasaas sai gsnigifincifaincatndte cdreecarseeaisne binlo obdlogoldu cgolsuecocosen cceonntcraetnitornastiboentsw beeetnw1eeann d1 2ahndp o2s th
cpoonsstu cmonpstiuomnpfotirotnh feocro tnhter ocolnantrdolh aignhd- hLiAghm-LeAal sm(ecoalnst r(coolnptr<o0l .p0 5<, 0h.i0g5h, -hLiAghp-L<A0 p.0 <5 )0,.t0h5o),u tghhoungoht fnoort
thfoer htihgeh -hOigAh-mOeAa lm. Deeasl.p Diteestphietsee tahletseer aatilotenrsa,ticoanlcsu, lcaatlicounlaotfioanre aofu anrdeear uthnedecru rtvhee (cAuUrvCe) (oAfUthCe) bolfo othde
gblulocoodse glleuvceolssei nletvheels2 inh tphoes 2t cho pnosustm cpotnisounmopfteiaocnh otfe setamche taelsfto muneadl nfoousnigdn nifioc saingtndifiifcfaenret ndcieffseirnenbcloeso din
gblulocoodse grluescpoosen sreesbpeotnwseee bnetthweedenif ftehree ndtiftfeesrtemnte taelsst (mdaetaalsn (odtastha onwotn s,hCoownntr,o CloAnUtrCol= A8U6C.2 4= ±865.2.245 ±A 5U.25,
hAigUh,- OhiAghA-OUAC A=U85C.2 =1 8±5.23.15 2± A3.5U2, AhiUgh, -hLigAhA-LUAC A=U8C5 .=0 885±.038. 5±3 3A.5U3 A(mUe (amne±anS E± MSE))M. )).
CChhaannggeesst otoi ninssuulilninc coonncceennttrraattiioonnss wweerreem meeaassuurreedda assp prreevvioiouussr reesseeaarrcchhi ninddicicaatetesst htheep pootetenntitaiall
mmoodduulalattioionno offp poosstptprraannddiaiallr reessppoonnsseessb byym moonnoouunnssaattuurraatteeddf faattss[ 4[400].].C Coonnssuummppttiioonno offa allllt etessttm meeaalsls
rreessuultleteddi nina as sigignnifiifcicaanntti ninccrreeaasseei nini ninssuulilnina attt hthee1 1h hp poossttc coonnssuummppttioionnt timimeep pooinintt( (ccoonnttrroollp p< < 00..0055,,
hhigighh-O-OAAp p< <0 0.0.055,,h hiigghh--LLAAp p< < 00..0055)),, aass sshhoowwnn iinn FFiigguurree5 5aa.. IInn rreessppoonnssee ttoo ccoonnssuummppttiioonno offt thheec coonnttrrooll
mmeeaal,l,i ninssuulilninl elevveelslsw weerrees stitlillls sigignnifiifcicaanntltylye elelevvaateteddc coommppaarereddt otob baasseelilnineea attt hthee2 2h hp poossttc coonnssuummpptitoionn
titmimeep pooinintt( p(p< <0 0..0055)).. IInn rreessppoonnssee ttoo tthhee ccoonnssuummppttiioonno offt thheeh hiigghh--LLAAt teessttm meeaall,,t thheerreew waassa as sigignnifiifcicaanntt
ddeeccrreeaassee (p(p< < 00.0.055)) iinn iinnssuulliinn ccoonncceennttrraattiioonnss bbeettwweeeenn tthhee 11 aanndd 22 hh ppoosstt ccoonnssuummppttiioonn ttiimmee ppooiinnttss..
DDeessppitietet hthisi,sn, onosi gsnigifincifaincatndti fdfeirffeenrceenscwese rweefroeu nfodubnedt wbeeetwnetheen ttehset mteesatl smdeuarlisn dgusruibnsge qsuuebnsetqnueetnAtU nCet
aAnaUlyCs iasn,aalsycsaisn, abse csaene nbein sFeeignu irne F5ibg.ure 5b.

Figure 5. Change in insulin over 2 h postprandial period. (a) plotted values. = control meal,
(cid:4)=high-OA,(cid:78)=high-LA;(b)Netareaundercurve(AUC)frombaseline.Measuredinplasmausing
● ■
aFmiguulrteip 5le. xCihmanmguen iona isnssauy.liAn lolvdeart a2 dhi sppolastyperdanadsimale paenr±iodS.E (Ma) ,palloltdteadta vpaoluinetss. n= =8 .co*n=trsoigl nmifieacal,n tly =
dhififgehr-eOnAt(,p ▲< =0 .h0i5g)ht-oLbAa;s (ebli)n Neefto arraelal tuensdtemr ecaulrsv.e# (=AUsigCn) iffircoamn tblyasdeilfinfeer.e Mnte(apsu<re0d.0 5in) tpolabsamseal iunseinfgo ra
cmonutlrtoiplmlexe ailm.†m=unsiogansisfiacya.n Atlyll ddiafftear ednistp(lpa<ye0d.0 a5s) tmoe1ahn f±o rStEhMe,h iagllh -dLaAta tepsotinmtse anl. = 8. * = significantly
different (p < 0.05) to baseline for all test meals. # = significantly different (p < 0.05) to baseline for
Gcohnretrloinl mweaals. †a = hsiogrnmifiocannetlyo fdiifnfetreernets (tp i<n 0.t0h5i)s tos t1u hd fyord thuee htioghp-LrAev tieosut mseraels. earch indicating that
postprandiallevelscanbemodulatedbythelevelofFAsaturation[28]. Inresponsetothehigh-LA
testmGeahlrse’lcionn swuams pat iohno,rtmheorneew oafs ainstiegrneisfit cainn ttihnicsr esatsued(yp <du0e.0 5t)oi npgrehvrieoliunsp rreosdeuacrtciho ninfrdoimcabtiansge litnhea,t
cpoomsptparraenddtioal tlheeveclosn ctarno lbme emaoldwuhlaetneda nbayl ythzien lgevthele onf eFtAA sUaCtu.rAatdiodnit [io28n]a. lIlny, rtehseproenwsea stoa thsieg hniigfihca-LnAt
rteedsut cmtioenalisn’ gcohnresluinmcpotniocnen, ttrhaetiroen wsfaosl loaw siinggnicfoicnasnutm inpctrioenasoef (tph e<c o0n.0t5ro) limn egahlraetlibno tphrtohdeu1ct(ipon< 0fr.0o5m)
abnadse2lihne(p, <co0m.0p5a)rteimd etop othinet scwonhteronl cmomeapla rwehdetno baansaelyliznienags tshheo wnent iAnUFiCg.u rAed6dai.tionally, there was a
significant reduction in ghrelin concentrations following consumption of the control meal at both the
1 (p < 0.05) and 2 h (p < 0.05) time points when compared to baseline as shown in Figure 6a.
The plasma concentration of the hormone GIP can be altered acutely by food intake [41]. In
response to consumption of all test meals there was a significant increase in GIP at the 1 h post
consumption time point (all p < 0.05), as shown in Figure 7a. Consumption of the control (p < 0.05)
and LA (p < 0.05) test meals resulted in a sustained significant difference when compared to baseline
at the 2 h post consumption time point, though not for the high-OA test meal. At the 2 h time point
Nutrients 2018, 10, x FOR PEER REVIEW 9 of 17
there was a significant decrease in GIP concentrations following consumption of the high-OA test
meal (p < 0.05), though there were no significant differences in net AUC between the different meals.
At 1 h post meal consumption, there was a significant increase (p < 0.05) in resistin following
consumption of the high-LA test meal, though this failed to result in a significant difference in net
Nutrients2018,10,1376 9of17
AUC for the test period (p = 0.13 compared to control), as shown in Figure 8b.
Nutrients 2018, 10, x FOR PEER REVIEW 9 of 17
there was a significant decrease in GIP concentrations following consumption of the high-OA test
meal (p < 0.05), though there were no significant differences in net AUC between the different meals.
At 1 h post meal consumption, there was a significant increase (p < 0.05) in resistin following
consumption of the high-LA test meal, though this failed to result in a significant difference in net
AUC for the test period (p = 0.13 compared to control), as shown in Figure 8b.

Figure 6. Change in ghrelin over 2 h postprandial period. (a) plotted values, = control meal,
(cid:4) =Fhiigguhr-eO 6A. C,h(cid:78)an=geh iing ghh-LreAli;n (obv)erN 2e ht pAoUstCprafrnodmial bpaesrieoldin. e(a.) pMloetatesdu rveadlueins, p●l a=s mcoanturosli nmgeaal, m■ul t=ip lex
immuhingoha-OssAa,y .▲A ll=d haitgahd-LiAsp; la(by)e dNaets AmUeaCn f±romSE bMas,ealilnled. aMtaepasouirnetds nin= p8la.s*m=a suigsinnigfi caa nmtluyltidpilfefxe rent
(p<0im.05m)uantotahsissatyi.m Aellp doaitnat dciospmlapyaerde dast ombeaans e±l iSnEeMfo, ralcl odnattrao plominetas ln. #= =8. s*i =g nsiigfinciafinctalnytldyi dffieffreernetn(t p(p< <0 .05)
tocon0.t0r5o)l amt tehails. time point compared to baseline for control meal. # = significantly different (p < 0.05) to
control meal.
The plasma concentration of the hormone GIP can be altered acutely by food intake [41].
InresponsetoconsumptionofalltestmealstherewasasignificantincreaseinGIPatthe1hpost
consumptiontimepoint(allp<0.05),asshowninFigure7a. Consumptionofthecontrol(p<0.05)
andLA(p<0.05)testmealsresultedinasustainedsignificantdifferencewhencomparedtobaseline

atthe2hpostconsumptiontimepoint,thoughnotforthehigh-OAtestmeal. Atthe2htimepoint
therewasFaigsuigren 6ifi. Cchaanntgde eincr geharseelinin ovGeIrP 2 cho pnocsetpnrtarnadtiiaol npserfiooldl.o (wa) ipnlgottceodn vsaulumesp, t●io n= coofnttrhoel mhiegahl, -■OA =t estmeal
(p<0.05),htihgoh-uOgAh, t▲he re= whiegrhe-LnAo; s(big) nNifietc aAnUtCd iffrfoemre nbcaesseliinne.n eMteAasUurCedb eint wpelaesnmtah uesdinigff ear emnutlmtipeleaxl s. At1h
postmealicmomnusnuomaspsatiyo. nA,llt hdeatrae dwisapslaayesdi gasn mifiecaann ±t SinEMcr,e aalsl eda(tpa <po0in.0ts5 )n i=n 8r. e* s=i sstiginniffioclalnotwly idnigffecroennts (upm < ptionof
0.05) at this time point compared to baseline for control meal. # = significantly different (p < 0.05) to
thehigh-LAtestmeal,thoughthisfailedtoresultinasignificantdifferenceinnetAUCforthetest
control meal.
period(p=0.13comparedtocontrol),asshowninFigure8b.

Figure 7. Change in glucose-dependent insulinotropic peptide (GIP) over 2 h postprandial period. (a)
plotted values, ● = control meal, ■ = high-OA, ▲ = high-LA; (b) Net AUC from baseline.
Measured in plasma using a multiplex immunoassay. All data displayed as mean ± SEM, all data
points n = 8. * = significantly different to baseline for all test meals. # = significantly different (p < 0.05)
to baseline for control, † = significantly different (p < 0.05) to baseline for high-LA test meal. ^ =
significantly different (p < 0.05) to 1 h for the high-OA test meal.

Figure7. Changeinglucose-dependentinsulinotropicpeptide(GIP)over2hpostprandialperiod.
(a)ploFtitgeudrev 7a.l uCehsa,nge= inco gnlutrcoolsme-deeapl,e(cid:4)nd=enht iignhsu-OlinAo,t(cid:78)rop=ich pigehp-tiLdAe ;(G(bIP))N oevteAr 2U hC pforsotmprabnadsiealli npee.riMode. a(sau) red
inplapslmotateud svinagluaesm, ●ul t=ip cleoxntirmolm muenaol,a s■say .=A hlligdha-OtaAd, is▲pl a=y ehdigahs-LmAe; a(nb)± NSeEt MAU,aCll fdroamta pboasineltisnen. =8.
*=significantlydifferenttobaselineforalltestmeals.#=significantlydifferent(p<0.05)tobaselinefor
Measured in plasma using a multiplex immunoassay. All data displayed as mean ± SEM, all data
control,†=significantlydifferent(p<0.05)tobaselineforhigh-LAtestmeal.ˆ=significantlydifferent
points n = 8. * = significantly different to baseline for all test meals. # = significantly different (p < 0.05)
(p<0t.o0 5b)atsoel1inhe ffoorr tchoenthriogl,h -†O =A sitgensitfimcaenatll.y different (p < 0.05) to baseline for high-LA test meal. ^ =
significantly different (p < 0.05) to 1 h for the high-OA test meal.
Nutrients 2018, 10, x FOR PEER REVIEW 10 of 17
Nutrients2018,10,1376 10of17
Nutrients 2018, 10, x FOR PEER REVIEW 10 of 17

Figure 8. Change in resistin over 2 h postprandial period. (a) plotted values, ● = control m eal, ■ =
Figure 8. Change in resistin over 2 h postprandial period. (a) plotted values, = control meal,
Fig(cid:4)huirg=eh h8-O.i gCAhh,- aO▲nAg,e (cid:78)=in h=reighshiisg-tLhinA- Lo; Av(e;br() b 2N )hNe pte otAstAUpUrCaC nfdrfioraomlm p ebbraiasosedeli.l ni(naee). .pMlMoetaeteasudsur veradeld uienins , pp●llaa =ssm mcoaan tuurssoiilnn mgg eaaa lm,m ■uullttii pp=ll eexx
higiimhm-mOmuAunn, oo▲aassss aa=yy ..h AAiglllhl d-dLaaAttaa; dd(ibiss)pp lNlaayyeete ddA aaUss Cmm efearanon m±± SbESaMEseM, lai,nllae .ld laMdtaae tapasoupinroetisdn tnsi n=n 8p=. l*a8,s .smi*ga,n siuifgiscnianinfigtc layan dtmliyfufedltriiefpnfleter x(ep n t<
(0p.0<50) .a0t5 t)haits tthimiset ipmoeinpto cionmtcpoamrepda rteod btaoseblainseel finore tfhoer tLhAe LmAeaml. eal.
immunoassay. All data displayed as mean ± SEM, all data points n = 8. *, significantly different (p <
0.05) at this time point compared to baseline for the LA meal.
AAss sshhoowwnn iinn FFiigguurree 99,, tthheerree wweerree nnoo ssiiggnniifificcaanntt cchhaannggeess iinn aaddiippoonneeccttiinn ccoonncceennttrraattiioonnss oorr
aaddiippoonneeccttiinn nneett AAUUCC ffoolllloowwiinngg ccoonnssuummppttiioonno offa annyyo offt thheet teessttm meeaalsls..
As shown in Figure 9, there were no significant changes in adiponectin concentrations or
adiponectin net AUC following consumption of any of the test meals.

Figure9. Changeinadiponectinover2hpostprandialperiod. (a)plottedvalues, =controlmeal,
(cid:4)Fig=uhreig 9h.- COhAa,n(cid:78)ge= inh aigdhip-LoAne;c(tbin) NoveetrA 2U hC pofrsotpmrabnadsiealli npee.riMode. a(sau) rpeldotitnedp vlaaslmueas,u ●si n= g caonmtruollt mipeleaxl,
Figi■murme u=9n . hoCiaghshas-naOyg.AeA ,i nl▲l ad da=itp ahodingisehpc-tlLianAy e;o dv(bea)rs N2m ehet a pAnoU±stCpS rEfarMnodm,iaa blll apdseaertliiaondpe..o i(Mna)te sapnslou=trte8ed.d ivna plulaess,m ●a u= scionngt rao ml muletaipl,l ex
■Tim h=em rheuignwho-aaOsssAna,yo .▲ sAi gl=ln dhifiaitgcaha d-nLitsApc;hl a(aybne) gdNe aetsto mAGeUaLCnP ±-f1r So,EmgMl ub,c aaaslegl ldoinanet,a. l Meppoetianinstus, rIneL d=- 18iβn. ,pIlLa-s6m,aIL u-s1i0n,gI La -m13u,lTtipNleFx-α ,orC
peptideinresponsetotestmealconsumption(seeFiguresS2–S10). MeasurementofPAI-1,visfatin,
immunoassay. All data displayed as mean ± SEM, all data points n = 8.
There was no significant change to GLP-1, glucagon, leptin, IL-1β, IL-6, IL-10, IL-13, TNF-α, or
IFN-γ, IL-2, IL-3, IL-4, IL-5, and IL-12p70 failed to yield complete data sets due to samples being
C peptide in response to test meal consumption (see Figures S2–S10). Measurement of PAI-1, visfatin,
outTshideeret hwearsa nngoe soigfndiefitcearnmt icnhaatinogne, aton dGaLsPs-1u,c ghluwceargeorne,m leopvteidn, fIrLo-m1βs,t aILti-s6t,i cIaLl-a1n0,a IlyLs-1is3., TNF-α, or
IFN-γ, IL-2, IL-3, IL-4, IL-5, and IL-12p70 failed to yield complete data sets due to samples being
C peptiTdhee irne rweseproennsoe stoig tneisfit cmaenatld cioffnesruemncpetsioonb s(seerev eFdigbuertews eSe2n–Sb1a0s)e. lMineeamsueraesmuernemt oefn PtsA(Ip-1r,e v-tiessfattmine, al
outside the range of determination, and as such were removed from statistical analysis.
IFNco-nγs, uImL-p2t, iIoLn-)3,f oILr-a4n, yILo-5f,t haneda nILal-y12teps70o rfaaiplepde ttiote ypiealrda mcoemteprslemte edaastuar esdet.s Nduoei mtop soarmtapnltesh abreminsgo r
There were no significant differences observed between baseline measurements (pre-test meal
ouutsniidnete tnhde erdanegffee cotfs dweeterremoibnsaetrivoend, .and as such were removed from statistical analysis.
consumption) for any of the analytes or appetite parameters measured. No important harms or
There were no significant differences observed between baseline measurements (pre-test meal
unintended effects were observed.
consumption) for any of the analytes or appetite parameters measured. No important harms or
unintended effects were observed.