method;agreementofgenotypedistributionwithHardy– Weinbergexpectationswasverifiedbyanexacttest.Link- /ott/eh.htm) (37). The differences in allele and frequencies between long-lived participants and regression. Comparison between the mean number

Table1. ESR1andCYP19GenotypeDistributionsinLong-Lived ParticipantsandControls

Table2. EstimatesoftheEffectsofESR1andCYP19Genotypeson ProbabilityofSurvivingtoAgeMoreThan90years

Controls LongLived ESR1genotypes  PP(%) 16(18.8) 32(29.6)  Pp(%) 51(60.0) 50(46.3)  pp(%) 18(21.2) 26(24.1)  N 85 108  XX(%) 17(20.0) 29(27.6)  Xx(%) 49(57.6) 56(53.3)  xx(%) 19(22.4) 20(19.0)  N 85 105  PPXX(%) 15(17.6) 29(27.6)  PPXx(%) 1(1.2) 3(2.9)  PpXX(%) 2(2.4) 0  PpXx(%) 44(51.8) 47(44.8)  Ppxx(%) 5(5.9) 3(2.9)  ppXx(%) 4(4.7) 6(5.7)  ppxx(%) 14(16.5) 17(16.2)  N 85 105 CYP19genotypes  GG(%) 36(45.0) 33(30.0)  GT(%) 36(45.0) 59(54.6)  TT(%) 8(10.0) 16(14.8)  N 80 108

Variables OR(95%CI) pValue Gender(male) 0.62(0.34–1.13) .12 ESR1PPgenotype 2.20(1.06–4.59) .035 CYP19Tcarryinggenotypes 1.92(1.04–3.55) .038

Note:CI=confidenceinterval;OR=oddsratio.

longlivedandthecontrols(PPXX:27.6%and17.6%,re- groups, the genotype frequencies of the CYP19 polymorphism agreed with those expected according Hardy–Weinbergequilibrium.Thers4646Gallelefrequen- cies in the long lived and in the controls were withthecontrols(p=.04).

from http://biomedgerontology.oxfordjournals.org/ at New York Medical College on November 16, 2014

AsbothESR1andCYP19areinvolvedinestrogenfunc- tion,tobetterevaluatetheiroverallactiononlongevityand tostudytheeffectofeachgenotypeadjustedfortheother variables,thedataweretestedbylogisticregression.Inthe analysis,thedependentvariablewashavingsurvivedtoage 90yearsandolder(≥90years)ornot,andtheexplanatory variableswereESR1andCYP19genotypesandgender.Asit hasbeenshownthattheESR1PvuIIpolymorphismmight modify ESR1 transcription and thus estrogen action (31), only ESR1 PvuII genotypes were included together with CYP19genotypesinaninitialmodel(Table2).Theanalysis indicatesthat,afteradjustingforgender,boththeESR1and the CYP19 polymorphisms may be independent determi- nants of longevity because the ESR1 PP and CYP19 T allele–carryinggenotypessignificantlyassociatedwithsur- vivingtomorethan90yearsofage(ESR1PPgenotype: adjustedOR=2.20andCYP19Tallele–carryinggenotypes: OR=1.92).Inasubsequentanalysis,similarresultswereob- tainedwhencombinedESR1PPXXwereconsidered(ESR1 PPXXgenotype:adjustedOR=2.17,95%confidenceinter- val:1.01–4.64,p=.046andCYP19Tallele–carryinggeno- types OR=1.92, 95% confidence interval: 1.02–3.56, p=.039).ESR1XXgenotypesseemtohavelessinfluence on survival (adjusted OR=1.84, 95% confidence interval: 0.88–3.81,p=.10).NosignificantinteractionofESR1and CYP19genotypeswithgenderwasobserved.

children associated with different CYP19 genotypes was performed using a nonparametric test (Kruskal–Wallis). Statistical analyses were performed using Statistix 8.0 AnalyticalSoftware.

Results

To evaluate the involvement of the ESR1 and CYP19 polymorphisms in longevity, ESR1 and CYP19 genotype frequencies observed in the long-lived participants were comparedwiththoseobservedintheparticipantswhohad diedbeforetheageof90years(controls).Theresultsare reportedinTable1.Inboththecontrolandthelong-lived groups,thegenotypefrequenciesofESR1PvuIIandXbaI polymorphisms agreed with those expected according to Hardy–Weinbergequilibrium.TheESR1*Pallelefrequen- ciesinthecontrolsandthelonglivedwere0.488and0.528, respectively.TheESR1*Xallelefrequenciesinthecontrols andinthelonglivedwere=0.488and0.543,respectively. BecauseoftheproximityofthetwoESR1RFLPs,thelink- agedisequilibriumbetweenthemwasanalyzed.Theywere instronglinkagedisequilibriuminbothsamples(p<.0001), withatendencyoftheESR1*Palleletobeassociatedwith ESR1*XandESR1*ptobeassociatedwithESR1*x.Conse- quently, as shown by the combined distribution of ESR1 genotypes(Table1),morethan90%ofESR1PPhomozy- gotesofbothgroupswereXXhomozygotesaswell.Com- parison of genotype distributions between the long lived andthecontrols(Table1)showsthattherewasanexcessof PP(29.6%vs.18.8%,p=.08)andXX(27.7%vs.20.0%, p=.22)genotypesinthelonglivedversusthecontrols,although thecomparisons(Pearson’sgoodness-of-fitchi-square)did

To investigate the involvement of the two genes in polymorphismsonpastfertilityofthesamesamplepopula- ppxxgenotypeswerefoundtobeassociatedwithanabove- among the men, whereas among the women, the

Table3.

isthattheinvolvementofthetwogenesinlongevitycould dependontheprotectionestrogensconferagainstagingby upregulatingtheexpressionofantioxidantlongevitygenes, suchasselenium-dependentglutathioneperoxidase(GPx) andMn-superoxidedismutase(Mn-SOD)(41).

Gene Genotype Number No.ofChildren CYP19rs4646 GG 58(0.37) 3.5±0.3 GT 78(0.49) 3.8±0.3 TT 22(0.14) 3.7±0.5

ESR1polymorphismsonbothlifespanandcompletefertil- tohavenoeffectontheactualnumberofchildrenapartici- pant had had. In contrast, the ESR1 genotypes we be associated with a reduced reproductive success in antagonistpleiotropy.Thesamepatternofantagonisticplei- otropywasobservedfortheinterleukin-10gene(16).Con- versely, the peroxisome proliferator–activated apositivepleiotropiceffectinagender-specificway(14), amixtureoftrade-offpatterns(14).

Note:Kruskal–Wallispvalue=.34.

reducedreproductiveefficiency.Inthepresentstudy,asimi- differenceinthenumberofchildrenamongthethreegeno- andnumberofspontaneousabortions(datanotreported).

Discussion

was to avoid the “cohort effect” observed in studies on human longevity when the control sample composed of younger participants belonging to birth co- environmentaleffectsthedistinctcohortsexperienced.Fur- “younger”controls,whoselifespanisunknown.

genes(andestrogen)areinvolvednotonlyinreproduction butalsoinagingandthattheymayexertapleiotropicaction acrossthewholecourseofhumanlife.Thepossibleimpor- tanceofthesestudiesisthattheymayhelptoelucidatethe agingprocessandtoidentifytheproperstrategiesforslow- ingand/ormitigatingitseffects.