2024年5月12日发(作者：习婷美)

NutritionalManipulationofPrimateRetinas,III:Effects

ofLuteinorZeaxanthinSupplementationonAdipose

TissueandRetinaofXanthophyll-FreeMonkeys

n,

1

MarthaNeuringer,

2,3,4

l,

1

WolfgangSchalch,

5

and

dderly

6,7,8

P

URPOSE

.Macularpigment(MP)iscomposedofthexantho-

phyllslutein(L)andzeaxanthin(Z)andmayhelptoprevent

thisstudytheeffectsofLorZsupplementationoncarotenoid

levelswasexaminedinserum,adiposetissue,andretinain

rhesusmonkeyswithnopreviousintakeofxanthophylls.

M

ETHODS

.Frombirthto7to16yearsofage,18rhesusmon-

keyswerefedsemipurifieddietscontainingallessentialnutri-

esupplementedwithpureL

and6withpureZat3.9

␮

mol/kgperdayfor24to101weeks.

Atbaselineandat4-to12-weekintervals,carotenoidsinadi-

ycompletion,

carotenoidsinserumandretina(central4mm,8-mmannulus,

andtheperiphery)swerecompared

withdatafromcontrolmonkeysfedastandardlaboratorydiet.

R

ESULTS

.Monkeysfedxanthophyll-freedietshadnoLorZin

orZsupplementation,serumand

adiposetissueconcentrationssignificantlyincreasedinthe

nd3R,3ЈS-Z(RSZormeso-Z,not

presentinthediet)wereincorporatedintoretinasofmonkeys

supplementedwithL,withRSZpresentonlyinthemacula

(central4mm).All-transZ,butnoRSZ,accumulatedinretinas

ofmonkeyssupplementedwithZ.

Fromthe

1

JeanMayerUSDAHumanNutritionResearchCenteron

AgingatTuftsUniversity,Boston,Massachusetts;the

2

Divisionof

Neuroscience,OregonNationalPrimateResearchCenterandDepart-

mentsof

3

Medicineand

4

Ophthalmology,OregonHealthandScience

University,Portland,Oregon;

5

DSMNutritionLtd.(formerlyRoche

VitaminsLtd.),Basel,Switzerland;and

6

SchepensEyeResearchInsti-

tute,DepartmentofOphthalmologyand

7

PrograminNeuroscience,

HarvardMedicalSchool,Boston,Massachusetts.

8

Presentaffiliation:DepartmentofOphthalmology,MedicalCol-

legeofGeorgia,Augusta,Georgia.

Anyopinion,findings,conclusions,orrecommendationsex-

pressedinthispublicationarethoseoftheauthorsanddonotneces-

sarilyreflmentofAgriculture.

SupportedbyDSMNutritionalProducts,Ltd.(formerlyRoche

VitaminsLtd.,Basel,Switzerland);Grant581950-9-001fromtheU.S.

DepartmentofAgriculture;GrantDK-29930fromtheInstituteof

DiabetesandDigestiveandKidneyDiseases(MN);GrantRR-00163

fromtheDivisionofResearchResources,NationalInstitutesofHealth;

andagrantfromTheFoundationFightingBlindness(MN).

SubmittedforpublicationNovember21,2002;revisedJuly1,

2003andJanuary26andOctober6,2004;acceptedOctober18,2004.

Disclosure:n,DSMNutritionalProducts,Ltd.(F);M.

Neuringer,DSMNutritionalProducts,Ltd.(F);l,DSM

NutritionalProducts,Ltd.(F);h,DSMNutritionalProducts,

Ltd.(E,F);rly,DSMNutritionalProducts,Ltd.(F)

Thepublicationcostsofthisarticleweredefrayedinpartbypage

ticlemustthereforebemarked“advertise-

ment”inaccordancewith18U.S.C.§1734solelytoindicatethisfact.

Correspondingauthor:n,JeanMayerUSDA

HumanNutritionResearchCenteronAgingatTuftsUniversity,711

WashingtonStreet,Boston,MA02111;n@.

692

C

ONCLUSIONS

.ListheprecursorofRSZ,amajorcomponentof

phyll-freemonkeyscanaccumulate

retinalxanthophyllsandprovideavaluablemodelforexamin-

ingtheiruptakeandconversion.(InvestOphthalmolVisSci.

2005;46:692–702)DOI:10.1167/iovs.02-1192

L

utein(L)andzeaxanthin(Z)arexanthophylls(carotenoids

thatcontainoneormorepolarfunctionalgroups)that

selectivelyaccumulateintheretinaandareparticularlydense

inthefovealregion,ormacula,wheretheyarethemain

componentsofthemacular

2,3

pigment.

1

LandZareknownto

functionasantioxidantsandblue-lightfiltersandthereby

mayprotectthemacularretinaandretinalpigmentepithelium

fromlight-initiatedoxidativedamage.

4

Recentstudiesinquail

exposedtobrightlightprovideevidencethatlong-termzeax-

anthinsupplementationleadstoincreasedretinalzeaxanthin

andreducedphotoreceptordeath.

5,6

Boneetal.

7

havestudiedtheretinaldistributionofLandZ

in

ϳ

-to-Zratioincreasedfroman

in

1:2.4

theperiphery

inthecentral

(8.7–12.2

macula

mm

(0–0.25

eccentricity).

mmeccentricity)

average

7,8

These

to

investi-

Ͼ2:1

of

gatorshaveshownthecomponentsofhumanmacularpigment

tobeL[(3R,3ЈR,6ЈR)-

␤

,

⑀

-carotene-3,3Јdiol];Z[3R,3ЈR)-

␤

,

␤

car-

otene-3,3

␤

♦

␤

-carotene-3,3

Јdiolor

Јdiol].

RRZ];

9

andRSZormeso-Z,3R,3ЈS-Z[(3R,3ЈS)-

RSZisprimarilylocatedinthecenterof

themacula,whereitisfoundtobeinanapproximateratioof

1:otgenerallypresentinthediet,but

probablyresultsfromchemicalprocessesoccurringwithinthe

eye.

9,10

Someinvestigatorshavespeculatedthattheoriginof

RSZisdietaryL.

9,10

However,untilnow,provingthissugges-

tionwasdifficult,giventhelackofbothanappropriateanimal

,primateswithnomacularpigment)andsufficient

quantitiesofpureLandZforcontrolledfeedingstudies.

Thereisepidemiologicevidencethatintakeoffoodshighin

Land/orZ,aswellashighserumlevelsofLϩZ,arerelatedto

reducedriskofadvancedage-relatedmaculardegeneration

(AMD)

11,12

(SanGiovanniJP,2004;45:ARVOE-abstract

2242).

Thisfindinghasraisedthequestionofwhether

supplementalintakeofLandZmaybeeffectiveinreducing

theriskforAMDorslowingitsprogression.

12–14

However,

therearemanygapsintheknowledgeabouttheuptakeand

metabolismofdietaryLandZandtheireffectivenessinraising

macularpigmentlevels.

Thepresentstudywasmadepossiblebytheexistenceofa

nimals

werefedsemipurifiedxanthophyll-freedietsfrombirthand

thereforehadnodetectablexanthophyllsinserumandlittleor

nomacularpigment,asmeasuredwithaninvivophotographic

method.

15

Thus,theeffectofdietarysupplementationwith

examinedtheeffectsofdietarysupplementationofthesexan-

urcesof

thesetwocarotenoidsweremadeavailabletousbyDSM

NutritionalProducts,Ltd.(formerlyRocheVitamins,Ltd.,

Basel,Switzerland)toallowcomparisonsbetweentheretinal

InvestigativeOphthalmology&VisualScience,February2005,Vol.46,No.2

Copyright©AssociationforResearchinVisionandOphthalmology

IOVS,February2005,Vol.46,No.2

T

ABLE

teristicsofMonkeys

Group,AnimalID

Xanthophyll-free

Mean

SEM

Zeaxanthin-fed

642

567

224

217

586

398

Mean

SEM

Lutein-fed

602

585

362

397

636

463

Mean

SEM

Control

Mean

SEM

Sex

2M,4F

F

F

F

F

M

M

XanthophyllAccumulationandTransformationinMonkeys693

Age

(y)

13.7

1.6

9.0

10.9

18.3

18.4

11.6

15.4

13.9

1.6

10.0

10.5

15.0

14.6

10.2

13.8

12.4

1.0

13.8

1.6

Bodywt

(kg)*

7.5

0.6

7.1

7.9

7.6

6.9

12.7

11.1

8.9

1.0

6.7

5.3

6.8

8.6

11.5

12.0

8.5

1.1

7.0

0.7

Supplement†

0

0

14(12,2)

14(12,2)

24(10,14)

24(10,14)

8(4,4)

8(4,4)

11(9,3)

2(2,1)

13(12,1)

13(12,1)

15(10,5)

15(10,5)

6(4,2)

6(4,2)

15(9,7)

3(2,2)

0

0

n-3FattyAcid

Status

4,low

2,adequate

Low

Low

Low

Adequate

Low

Adequate

F

F

F

F

M

M

2M,12F

Low

Low

Low

Adequate

Low

Adequate

Adequate

*Bodyweightatstudy’send.

†Numbersinparenthesesindicatedurationofsupplementation(inmonths)at7days/wkand4

days/wk,respectively.

orpapersinthisseriesdescribe

thetimecourseofincreasesinserumxanthophyllsandmacu-

larpigmentopticaldensityinvivo,

15

morphologicchangesin

theretinalpigmentepithelium,

16

andtheeffectsofacute

resentpaper,wereportlongitu-

dinalmeasuresofadiposetissuexanthophyllsandtheanalysis

ofserumandretinalcarotenoidsattheendofsupplementa-

tion,includingthelevelsofLandZandtheirmetabolitesinthe

udyprovidedauniqueopportu-

nitytodeterminetheeffectivenessofpureLandZinincreas-

ingmacularpigmentandtoidentifythedietaryoriginofRSZ.

M

ETHODS

AnimalsandDiets

AllprocedureswereapprovedbytheInstitutionalAnimalCareandUse

CommitteeoftheOregonNationalPrimateResearchCenterandcon-

formedtoNIHguidelinesandtheARVOStatementfortheUseof

enrhesusmonkeys

(Macacamulatta)wererearedononeoftwosemipurifieddiets,both

ofwhichcontainedadequatelevelsofallknownnutrients,including

vitaminA(asvitaminAacetate)and

␣

-tocopherol,butnodetectable

xanthophylls,ribedinmore

detailinNeuringeretal.,

15

thetwodietsdifferedonlyintheirfat

sourcesandthereforeinfattyacidcomposition,withonecontaining

lowlevelsandoneadequatelevelsofn-3fattyacidsintheformof

␣

-enodifferencesrelatedtofattyacidstatuswere

foundinanyoftheanalysesreportedherein,thedataforthesetwodiet

malsalsoreceivedlimitedamounts

ofverylowxanthophyllfoodssuchaswheatorricecereals,whiterice,

sweeteneddrinks,gelatin,pineapple,andbanana.

Beginningat7to16yearsofage,thedietsofsixofthesemonkeys

weresupplementedwithpureLandsixwithpureZ,at3.9

␮

mol/kg

perday(2.2mg/kgperday).Thisdoserepresented7.7timesthe

averagedailyxanthophyllintakefromthestandardlaboratorydiet

(describedlater).TheLandZsupplementswerepurifiedorsynthe-

sizedbyDSMNutritionalProducts,Ltd.(formerlyRocheVitaminsLtd.,

Basel,Switzerland)f

beadletsweredeterminedforeachanimalbasedonouranalysisof

plementswere

insertedintomarshmallows,sweetenedgelatin,orsmallpiecesoffruit.

Beadletsandindividualsupplementdoseswerestoredat4°Cinthe

mentswereprovideddailyfor4to12monthsand,dueto

limitedsupplyofthepurexanthophylls,fourtimesperweekthereafter

ationofdailysupplementa-

tionvariedwithineachgroupbutwasmatchedbetweenthetwo

r,becauseLhadtobespeciallypurified,itssupplywas

morelimited,resultinginashorterdurationofsupplementationatfour

timesperweekfortheL-fedgroup(3Ϯ1monthcomparedto7Ϯ2

monthsfortheZ-fedgroup).TheL-andZ-fedgroupswerebalancedto

theextentpossiblebasedonsex,n-3dietgroup,

remainingsixanimalscontinuedontheirsemipurifieddietsbutre-

ceivednoxanthophyllsupplements(xanthophyll-free).TwoL-fedand

twoZ-fedanimalswerekilledafter6to8monthsofsupplementation,

twofromeachgroupat13to14monthsandtwofromeachgroup

eachat15to24months(seeTable1fordetails).

DatafromtheL-andZ-fedmonkeyswerecomparedwithdatafrom

normalcontrolmonkeysfedastandardstockdiet(Purina5047Mon-

keyChow;RalstonPurina,Richmond,IN)providingadailycarotenoid

intakeof0.26

␮

mol/kgperdayL,0.24

␮

mol/kgperdayZ,and0.035

␮

mol/kgperday

␤

-carotene(meansoffouranalyses).Theseanimals

alsoreceivedsupplementalfruitsandvegetables(typically,onefourth

toonehalfanapple,oronehalfacarrotapproximatelythreetimesper

week),whichcontributedanestimatedmaximumadditionaldaily

averageofϳ3nmol/kgofLplusZorϽ1%oftheintakefromthestock

diet.

17

Theywerehousedunderthesameconditionsastheexperi-

sanalyzedfromcontrolmonkeysincluded

ltissuewas

analyzedfrom14controlmonkeys(2male,12female)including7for

4-mmmacularsamples,11for8-mmannularsamples,and8forpe-

ripheralsamples(seedescriptionofretinaltissuedissectioninthenext

section).

2024年5月12日发(作者：习婷美)

NutritionalManipulationofPrimateRetinas,III:Effects

ofLuteinorZeaxanthinSupplementationonAdipose

TissueandRetinaofXanthophyll-FreeMonkeys

n,

1

MarthaNeuringer,

2,3,4

l,

1

WolfgangSchalch,

5

and

dderly

6,7,8

P

URPOSE

.Macularpigment(MP)iscomposedofthexantho-

phyllslutein(L)andzeaxanthin(Z)andmayhelptoprevent

thisstudytheeffectsofLorZsupplementationoncarotenoid

levelswasexaminedinserum,adiposetissue,andretinain

rhesusmonkeyswithnopreviousintakeofxanthophylls.

M

ETHODS

.Frombirthto7to16yearsofage,18rhesusmon-

keyswerefedsemipurifieddietscontainingallessentialnutri-

esupplementedwithpureL

and6withpureZat3.9

␮

mol/kgperdayfor24to101weeks.

Atbaselineandat4-to12-weekintervals,carotenoidsinadi-

ycompletion,

carotenoidsinserumandretina(central4mm,8-mmannulus,

andtheperiphery)swerecompared

withdatafromcontrolmonkeysfedastandardlaboratorydiet.

R

ESULTS

.Monkeysfedxanthophyll-freedietshadnoLorZin

orZsupplementation,serumand

adiposetissueconcentrationssignificantlyincreasedinthe

nd3R,3ЈS-Z(RSZormeso-Z,not

presentinthediet)wereincorporatedintoretinasofmonkeys

supplementedwithL,withRSZpresentonlyinthemacula

(central4mm).All-transZ,butnoRSZ,accumulatedinretinas

ofmonkeyssupplementedwithZ.

Fromthe

1

JeanMayerUSDAHumanNutritionResearchCenteron

AgingatTuftsUniversity,Boston,Massachusetts;the

2

Divisionof

Neuroscience,OregonNationalPrimateResearchCenterandDepart-

mentsof

3

Medicineand

4

Ophthalmology,OregonHealthandScience

University,Portland,Oregon;

5

DSMNutritionLtd.(formerlyRoche

VitaminsLtd.),Basel,Switzerland;and

6

SchepensEyeResearchInsti-

tute,DepartmentofOphthalmologyand

7

PrograminNeuroscience,

HarvardMedicalSchool,Boston,Massachusetts.

8

Presentaffiliation:DepartmentofOphthalmology,MedicalCol-

legeofGeorgia,Augusta,Georgia.

Anyopinion,findings,conclusions,orrecommendationsex-

pressedinthispublicationarethoseoftheauthorsanddonotneces-

sarilyreflmentofAgriculture.

SupportedbyDSMNutritionalProducts,Ltd.(formerlyRoche

VitaminsLtd.,Basel,Switzerland);Grant581950-9-001fromtheU.S.

DepartmentofAgriculture;GrantDK-29930fromtheInstituteof

DiabetesandDigestiveandKidneyDiseases(MN);GrantRR-00163

fromtheDivisionofResearchResources,NationalInstitutesofHealth;

andagrantfromTheFoundationFightingBlindness(MN).

SubmittedforpublicationNovember21,2002;revisedJuly1,

2003andJanuary26andOctober6,2004;acceptedOctober18,2004.

Disclosure:n,DSMNutritionalProducts,Ltd.(F);M.

Neuringer,DSMNutritionalProducts,Ltd.(F);l,DSM

NutritionalProducts,Ltd.(F);h,DSMNutritionalProducts,

Ltd.(E,F);rly,DSMNutritionalProducts,Ltd.(F)

Thepublicationcostsofthisarticleweredefrayedinpartbypage

ticlemustthereforebemarked“advertise-

ment”inaccordancewith18U.S.C.§1734solelytoindicatethisfact.

Correspondingauthor:n,JeanMayerUSDA

HumanNutritionResearchCenteronAgingatTuftsUniversity,711

WashingtonStreet,Boston,MA02111;n@.

692

C

ONCLUSIONS

.ListheprecursorofRSZ,amajorcomponentof

phyll-freemonkeyscanaccumulate

retinalxanthophyllsandprovideavaluablemodelforexamin-

ingtheiruptakeandconversion.(InvestOphthalmolVisSci.

2005;46:692–702)DOI:10.1167/iovs.02-1192

L

utein(L)andzeaxanthin(Z)arexanthophylls(carotenoids

thatcontainoneormorepolarfunctionalgroups)that

selectivelyaccumulateintheretinaandareparticularlydense

inthefovealregion,ormacula,wheretheyarethemain

componentsofthemacular

2,3

pigment.

1

LandZareknownto

functionasantioxidantsandblue-lightfiltersandthereby

mayprotectthemacularretinaandretinalpigmentepithelium

fromlight-initiatedoxidativedamage.

4

Recentstudiesinquail

exposedtobrightlightprovideevidencethatlong-termzeax-

anthinsupplementationleadstoincreasedretinalzeaxanthin

andreducedphotoreceptordeath.

5,6

Boneetal.

7

havestudiedtheretinaldistributionofLandZ

in

ϳ

-to-Zratioincreasedfroman

in

1:2.4

theperiphery

inthecentral

(8.7–12.2

macula

mm

(0–0.25

eccentricity).

mmeccentricity)

average

7,8

These

to

investi-

Ͼ2:1

of

gatorshaveshownthecomponentsofhumanmacularpigment

tobeL[(3R,3ЈR,6ЈR)-

␤

,

⑀

-carotene-3,3Јdiol];Z[3R,3ЈR)-

␤

,

␤

car-

otene-3,3

␤

♦

␤

-carotene-3,3

Јdiolor

Јdiol].

RRZ];

9

andRSZormeso-Z,3R,3ЈS-Z[(3R,3ЈS)-

RSZisprimarilylocatedinthecenterof

themacula,whereitisfoundtobeinanapproximateratioof

1:otgenerallypresentinthediet,but

probablyresultsfromchemicalprocessesoccurringwithinthe

eye.

9,10

Someinvestigatorshavespeculatedthattheoriginof

RSZisdietaryL.

9,10

However,untilnow,provingthissugges-

tionwasdifficult,giventhelackofbothanappropriateanimal

,primateswithnomacularpigment)andsufficient

quantitiesofpureLandZforcontrolledfeedingstudies.

Thereisepidemiologicevidencethatintakeoffoodshighin

Land/orZ,aswellashighserumlevelsofLϩZ,arerelatedto

reducedriskofadvancedage-relatedmaculardegeneration

(AMD)

11,12

(SanGiovanniJP,2004;45:ARVOE-abstract

2242).

Thisfindinghasraisedthequestionofwhether

supplementalintakeofLandZmaybeeffectiveinreducing

theriskforAMDorslowingitsprogression.

12–14

However,

therearemanygapsintheknowledgeabouttheuptakeand

metabolismofdietaryLandZandtheireffectivenessinraising

macularpigmentlevels.

Thepresentstudywasmadepossiblebytheexistenceofa

nimals

werefedsemipurifiedxanthophyll-freedietsfrombirthand

thereforehadnodetectablexanthophyllsinserumandlittleor

nomacularpigment,asmeasuredwithaninvivophotographic

method.

15

Thus,theeffectofdietarysupplementationwith

examinedtheeffectsofdietarysupplementationofthesexan-

urcesof

thesetwocarotenoidsweremadeavailabletousbyDSM

NutritionalProducts,Ltd.(formerlyRocheVitamins,Ltd.,

Basel,Switzerland)toallowcomparisonsbetweentheretinal

InvestigativeOphthalmology&VisualScience,February2005,Vol.46,No.2

Copyright©AssociationforResearchinVisionandOphthalmology

IOVS,February2005,Vol.46,No.2

T

ABLE

teristicsofMonkeys

Group,AnimalID

Xanthophyll-free

Mean

SEM

Zeaxanthin-fed

642

567

224

217

586

398

Mean

SEM

Lutein-fed

602

585

362

397

636

463

Mean

SEM

Control

Mean

SEM

Sex

2M,4F

F

F

F

F

M

M

XanthophyllAccumulationandTransformationinMonkeys693

Age

(y)

13.7

1.6

9.0

10.9

18.3

18.4

11.6

15.4

13.9

1.6

10.0

10.5

15.0

14.6

10.2

13.8

12.4

1.0

13.8

1.6

Bodywt

(kg)*

7.5

0.6

7.1

7.9

7.6

6.9

12.7

11.1

8.9

1.0

6.7

5.3

6.8

8.6

11.5

12.0

8.5

1.1

7.0

0.7

Supplement†

0

0

14(12,2)

14(12,2)

24(10,14)

24(10,14)

8(4,4)

8(4,4)

11(9,3)

2(2,1)

13(12,1)

13(12,1)

15(10,5)

15(10,5)

6(4,2)

6(4,2)

15(9,7)

3(2,2)

0

0

n-3FattyAcid

Status

4,low

2,adequate

Low

Low

Low

Adequate

Low

Adequate

F

F

F

F

M

M

2M,12F

Low

Low

Low

Adequate

Low

Adequate

Adequate

*Bodyweightatstudy’send.

†Numbersinparenthesesindicatedurationofsupplementation(inmonths)at7days/wkand4

days/wk,respectively.

orpapersinthisseriesdescribe

thetimecourseofincreasesinserumxanthophyllsandmacu-

larpigmentopticaldensityinvivo,

15

morphologicchangesin

theretinalpigmentepithelium,

16

andtheeffectsofacute

resentpaper,wereportlongitu-

dinalmeasuresofadiposetissuexanthophyllsandtheanalysis

ofserumandretinalcarotenoidsattheendofsupplementa-

tion,includingthelevelsofLandZandtheirmetabolitesinthe

udyprovidedauniqueopportu-

nitytodeterminetheeffectivenessofpureLandZinincreas-

ingmacularpigmentandtoidentifythedietaryoriginofRSZ.

M

ETHODS

AnimalsandDiets

AllprocedureswereapprovedbytheInstitutionalAnimalCareandUse

CommitteeoftheOregonNationalPrimateResearchCenterandcon-

formedtoNIHguidelinesandtheARVOStatementfortheUseof

enrhesusmonkeys

(Macacamulatta)wererearedononeoftwosemipurifieddiets,both

ofwhichcontainedadequatelevelsofallknownnutrients,including

vitaminA(asvitaminAacetate)and

␣

-tocopherol,butnodetectable

xanthophylls,ribedinmore

detailinNeuringeretal.,

15

thetwodietsdifferedonlyintheirfat

sourcesandthereforeinfattyacidcomposition,withonecontaining

lowlevelsandoneadequatelevelsofn-3fattyacidsintheformof

␣

-enodifferencesrelatedtofattyacidstatuswere

foundinanyoftheanalysesreportedherein,thedataforthesetwodiet

malsalsoreceivedlimitedamounts

ofverylowxanthophyllfoodssuchaswheatorricecereals,whiterice,

sweeteneddrinks,gelatin,pineapple,andbanana.

Beginningat7to16yearsofage,thedietsofsixofthesemonkeys

weresupplementedwithpureLandsixwithpureZ,at3.9

␮

mol/kg

perday(2.2mg/kgperday).Thisdoserepresented7.7timesthe

averagedailyxanthophyllintakefromthestandardlaboratorydiet

(describedlater).TheLandZsupplementswerepurifiedorsynthe-

sizedbyDSMNutritionalProducts,Ltd.(formerlyRocheVitaminsLtd.,

Basel,Switzerland)f

beadletsweredeterminedforeachanimalbasedonouranalysisof

plementswere

insertedintomarshmallows,sweetenedgelatin,orsmallpiecesoffruit.

Beadletsandindividualsupplementdoseswerestoredat4°Cinthe

mentswereprovideddailyfor4to12monthsand,dueto

limitedsupplyofthepurexanthophylls,fourtimesperweekthereafter

ationofdailysupplementa-

tionvariedwithineachgroupbutwasmatchedbetweenthetwo

r,becauseLhadtobespeciallypurified,itssupplywas

morelimited,resultinginashorterdurationofsupplementationatfour

timesperweekfortheL-fedgroup(3Ϯ1monthcomparedto7Ϯ2

monthsfortheZ-fedgroup).TheL-andZ-fedgroupswerebalancedto

theextentpossiblebasedonsex,n-3dietgroup,

remainingsixanimalscontinuedontheirsemipurifieddietsbutre-

ceivednoxanthophyllsupplements(xanthophyll-free).TwoL-fedand

twoZ-fedanimalswerekilledafter6to8monthsofsupplementation,

twofromeachgroupat13to14monthsandtwofromeachgroup

eachat15to24months(seeTable1fordetails).

DatafromtheL-andZ-fedmonkeyswerecomparedwithdatafrom

normalcontrolmonkeysfedastandardstockdiet(Purina5047Mon-

keyChow;RalstonPurina,Richmond,IN)providingadailycarotenoid

intakeof0.26

␮

mol/kgperdayL,0.24

␮

mol/kgperdayZ,and0.035

␮

mol/kgperday

␤

-carotene(meansoffouranalyses).Theseanimals

alsoreceivedsupplementalfruitsandvegetables(typically,onefourth

toonehalfanapple,oronehalfacarrotapproximatelythreetimesper

week),whichcontributedanestimatedmaximumadditionaldaily

averageofϳ3nmol/kgofLplusZorϽ1%oftheintakefromthestock

diet.

17

Theywerehousedunderthesameconditionsastheexperi-

sanalyzedfromcontrolmonkeysincluded

ltissuewas

analyzedfrom14controlmonkeys(2male,12female)including7for

4-mmmacularsamples,11for8-mmannularsamples,and8forpe-

ripheralsamples(seedescriptionofretinaltissuedissectioninthenext

section).