, Materials ; ; and | | methods

$,2, Samples

southern | ee SNItaly | Was ¢ ; almost 1 | the s : same | : (about : 0F7 ““r*) In ¢ | centenarians < wesand « | controls. ' . The : : sample (cid:11)* ENDED DIDI OS‘centenarians ; comprised | | 2191 meni Maeunrelated : eu MEM| subjects | 6A(cid:30) males ; and (cid:31)DA females) « ) older | “eethan (cid:31)00 ) years, « , and i EEE TEE| included | all t | those | 6(cid:31)3@7 — een) analyzed i  lint Aa a\ the | : previous ; study « has | for’ SF ENN NE LE‘ TH/longevity SO Gh ENOassociation 6$(cid:2) Benedictis et (cid:17)(cid:10)!& (cid:31)3327!  we EE Iyounger   ON IEEcomprised  25 @ CEO EE Iunrelated subjects | 6(cid:31)(cid:31)3 | males | and (cid:31)/A EE EDfemales, | (cid:30)0(cid:23)A0 =“ Jeyears | (cid:11)* —_ erage) | vb Nene| randomly | ‘retrieved - *(cid:21)(cid:11)" _ the eecontrol me Mae| group ( 6/ 2 } subjects) « (cid:11)* F the | we ee: previous DNL I OE IE ES; above-cited s “ows eee| study. * . The : veg ee> Same » study | provides  SEEN” &detailed | oer Awe| description « (cid:11)* Ine“the « epee |criteria « (cid:11)* recruitment. . All | the : ey ee- subjects | Were | free | (cid:11)* revereclinically overt | EN OEEt pathologies. | In} 1 particular, t , the « > centenarians | ; were I in ¢ af es\ fairly | / good | | health s | Status < sand « oe)| did not peesuffer “oemajor  rspathologies  or disabilities (categories > and ) (cid:11)*  the classification reported in aes peeFranceschi | weeet | oNal. 6(cid:30)000(cid:19)77! All wee AXthe eossubjects had been invited to take part in oP”the | study | by 3 an | | appropriate |  KE EOcampaign ; feouee. and | [had | oo I| given | their | informed « | consent.

$,$, (cid:1)#( analyses

$:> . Was ¢ extracted *(cid:21)(cid:11)" | blood | | buffy. coats | following : - standard | | procedures. | _INS-FokI- ((cid:29)%9 -(Lucassen | et (cid:17)(cid:10)!& (cid:31)33/7 and NF OOD II IID II IIISIGF2-AvalI-RFLP tO IE(Schneid “et (cid:17)(cid:10)!& (cid:31)3237& which lie ame eee Oe eeedownstream « (cid:11)* the | eee-INS | 2 ea) gene wsand {in 1 \ the « - exon ! 3 (cid:11)* eerIGF2 | eee) gene, | NEN NE OOD A Arespectively, '  | were | - analyzed by restriction analysis on safter $:> DEEN ERIE ADIL IE ESPCR-amplification. SNE AMPPrimers beneand 9.( NN ESE! YE ISEOAEexperimental EEconditions | » are | NEE POLEavailable | / On | | request | NN OI OD IDI IIA II ILL INE. (g.debenedictis @unical.it).

Table :  

IGFII-Avall ((cid:29)%9 | genotypic | and | allelic | counts | and frequencies | 6(cid:29)(cid:21)(cid:2);7 in (cid:30)0(cid:23)A0 | year. old»   and in centenarians  >B9 > indicates Pe Oe ON Ees absence/presence « (cid:11)* F Avall 1 5 alk eee[ restriction s wee1 site

(cid:30)0(cid:23)A0  ) year «  r old s | subjects Centenarians  Males Females Males Females Counts  Freq Count  Freq Count  Freq Count  Freq Genotype >> D3 0!D(cid:31)(cid:30) @ 0!DAD // 0!D 2 @ 0!/2(cid:31) >9 @(cid:30) 0! 521 0!D0(cid:31) / 0!D2@ A/ 0!D3A 99 2 0!@A (cid:31)A 0!(cid:31)(cid:30)D D 0! @ (cid:31)2 0.122 Allele > (cid:31)@0 0!@A(cid:30) (cid:31)2 0!@A 101 0!A0(cid:31) (cid:31)2 0!@(cid:30)3 9 A2 0!/(cid:30)2 23 0!/(cid:30) D/ 0!(cid:30)33 (cid:31)03 0!/A(cid:31)

$,(cid:25), Statistical | analyses

For single markers, allele frequencies were computed by counting genes *(cid:21)(cid:11)" the observed | COE EEE| genotypes | wag SMsand | | exact | “eetests) were « - applied |  ———————) verify | BMF EEE IIS OE Hardy—Weinberg « - equilibrium 6(cid:22)-=7 6-(cid:2)(cid:8)(cid:21)& (cid:31)33@7! Homogeneity (cid:11)* “Pree oNgenotypic os Jdand yooallelic a —frequency Sp Se eedistributions en Mt NXbetween : branes”| sample | AMET- groups | , Was | oetested | ae| using | al Sees J ee| chi-square | statistics.

between pairs | (cid:11)* “markers  and to ACStest OMEthe    hypothesis     0(cid:1) oN eedMaximum  Likelihood ee EEEstimation | 6#%=7 | Was | used | to. estimate | ~—as I- haplotypic - frequencies.  Homogeneity | (cid:11)* haplo- typic | sewn Mannefrequency « ap NN eeedistributions | ; between : — Pe| sample | eT Pew- groups | —_—: Was | tested | by NE Eo/ means ¢ (cid:11)* ae MA eee’ Arlequin1.1 “J Pew feetsoftware | pe Fe(exact | tests | (cid:11)* population « NEE OIIII LEIS EIEN| differentiation).

 _ Results