2*<* Statistical | analysis

Fisher’s « exact | | probability 1 tests ; were | - performed 1 | to: ) ASSESS | ; whether 1 the | - frequency «  differ- ences observed eeebetween ae”the aged aim |and | hee—e—Xcontrol groups for haplotypes, ~oweengroups | eeof “related haplotypes : sand t | the 1 (cid:17)(cid:3)%+,:B(cid:17)(cid:3);(%% NE IEE DIDIER II) polymorphisms ' peaswere $ ETE I- statistically s Ss of re ™significant | fusing 1 r the eee Icomputer ‘ package - ‘microstat’. ' a |The | reine/ males ; “heeand | | females \ ; within | -each | en — ie| group | ) were | NSalso “oe indi- eeevidually © Pees onassessed for statistically significant frequency nee ENE Emdifferences “pe esusing the same —)approach. Multiple comparisons oe“oewere ann eewenestaken into Spee J”account by employing ONBonferroni’s  “Preecorrection, oe} + = ( ((cid:1);%+(cid:2)(cid:7) (where  is equal to the eeenumber J ™“of eee Pes FEN Oecomparisons), and the level | of | 7 rls* statistical Me Au| significance | adjusted ; eee| accordingly.

2*0* Phylogenetic analysis

ns waanalysed 1 wd ON”| using | / maximum | EEE EEE Ee| parsimony « criteria. ' This: eeewas | VJ bee eee; performed | | by 1 using | the « eycluster | of EE PONEprograms | in a —the | | PHYLIP | ee ipackage | ND ENN(Version | 4(cid:28)%(cid:5)) oe pee-Felstenstein, +;;4>(cid:28) J eee =>First, | the 1 restriction resomatrix | was | EIN ED EOE EP; bootstrapped +((( SoM VT) times | » by 1 using “wp Athe discrete ED A A A”- character « xen ney‘ option i in | . the : seqboot program. | NE IOS. Maximum | rPree™ eee| parsimony 1 trees | sys using | ae **the’ Wagner | ‘method | | were 1 “Peeweethen | DO IEE EN OST| generated { eee| from eeethis | oo"data by using oe pethe ee IIDdefault option in a —the oe OE Se aemixprogram. Finally,  NEE EOEconsensus tree was obtained, | Foousing | am *the |  CONSENSE | ae Spee- program.

2*(cid:30)* Analysis |  (cid:18)(cid:4)<4(cid:30)7 and | (cid:18)(cid:4)(cid:29)3<< | polymorphisms

French .and | | Japanese DEE IEE NINE IIS EE» centenarians, pee, the +(( pene) control s | samples ¢ sand t | the +’; ) aged : | samples \ > were amplified t | by | / OR 1 using | Se RE DREN MEEspecifically « designed | OE OEY| primers | pee; for t ‘the t > two! aA oO) regions ~—*; of I pTinterest, |  + meenand | ree| Mi‘ ’ “Msfor | the 1 (cid:17)(cid:3)%+,: “Peeregion —)and | | Mi | 4 seeand | ree| Mi « * | for | ‘the 1 (cid:17)(cid:3);(%% “oes) region | | (Table +>(cid:28) / oR amplification | 1 for « reach 1 Fi Ao”| region | 1 was ¢ y again | | performed i | in +(( ) pls ee MAM MAME| reaction ' “one KA. volumes ¢ AS ea pe es; outlined Meee pen”above | with | essentially 1 the : » Same « mee Sohcycling « Se: conditions « “ae eeexcept 1 eo: the ; annealing ; temperature » Was

increased | | to ( 3((cid:2) and 1 | the « - extension | | time 1 : reduced 1 | to + min. 1 . Amplification | | Was ° verified |  += agarose | gel « | electrophoresis | | stained | | with | _ ethidium | DELIA EDIT| bromide | <((cid:28)% ) g/ml).

SEEN ™ &Amplified ] o_rn*-e—~—| DNA ( <’ arepl) 1 ) was ; applied 1 I to ¢ pac i charged 1 {nylon 1 IIIA NID OE| membrane < ~ eee> aS | Olen Ee; previously 1 / reported Ee(Middleton « et i cal., +;;%>(cid:28) Jue “Pree_ Hybridisations, : we EEE Ee_ stringent | BE POEL OE| washes yand « IN ONE EE EEN FE| chemiluminescence « ———e—e—e—e: detec- YeeueVGue~tion of the ED ED A A |hybridised ENdigoxigenin IID ONE IID EON(DIG)-labelled probes were carried out as recently reported EN EF OLIN(Williams et moOal., +;;;>(cid:28) ae NRAStringent wash © peetemperatures and concentrations ae Ddfor apeeach of the probes wereoo 9 tA AA eeeempirically oN eeedetermined aneand eeare indicated in Table ’(cid:30) Mito +  4 } for 1 (cid:17)(cid:3);(%% and | | Mito : ’ rb”+. * ed| for 1 (cid:17)(cid:3)%+,:(cid:28) Int | the « > Case ( of 1 the 1 (cid:17)(cid:3)%+,:(cid:14) CAN ys aoa“. polymorphism, 4( ) DNA : EER NE. samples « ; derived | | from , | Asian 1 | individuals \ >; were ¢ also i y included i | in « mes Pe| order | “to. oN pee) verify t “eeethe results, Fe eeesince | re | positive - - hybridisation | result | for | this | ; polymorphism - | Was |} | not | found | aedin| | the Irish | ee RENN| population.

 , Results

/*4* Forty-six | /mtDNA | haplotypes