)*+* . Ross ¢ yet ¢ tal. / , / Experimental ( | Gerontology - /0 123345 4404644(cid:30)7

ED EEE ELpolymorphisms s within « Nr *1 our ¢ ‘oe 8r cohort « t of « f control ¢ | and « | aged « | cases ¢ eesand | | permit t t the « EO EEN> construction « 1 of  ee Peetable “Reeof genetic haplotypes incorporating aM SOthe —_S eevarious ENE EEE EEpolymorphic restriction sites “eR SSencountered, \ iinwhen | coop. bands « Vs J Peo; deviated 1 oe ee| from t \ the | ie MeN Teepredicted | “ieee pe| patterns i en J Oe ees; inferred | | from t 1 the - Anderson sequence ( OD NN ENED OE®- (Anderson ¢ Let tal., +;:+>(cid:28) . Such ; 1 an ¢ ~ eee| approach | ~_“ Pee. would | | facilitate « aC | comparative - analy- eetsis imo—™that “vr erewould ONE NEE NAN BEYdetermine a fan he Mewhether  pe OE OEEspecific  ree NE OShaplotype/group | of “related epee eeehaplotypes _ was more < > associated | with \ the aged « EEE RB{ community t than t np1 the ¢ swe PMS> control § mur NS| group. | . Secondly, i “hb, it ¢ pas J Pee”t contains s the +3(3%(cid:21) ‘ polymorphic 1 ——— ooorestriction : site, nee uewhich « | defines the | we Ew sre *- European . & EINE J AM| haplogroup ¢ y and 1 | would “Ns © b*therefore < ped ee> allow sus s tO ¢ ) determine | > whether t r the J & | haplogroup i ) is ¢ treealso i “ee EEE) increased i Lin « vb1 Our ¢ raged | Irish population. ’ _ Thirdly, | the | - absence of 1 ENE IN IIE ED‘recombination | or| with 3 yp ae eee| respect | to! ) the 1 » mtDNA : ee—oou_infers | that rerany « other CEE ALLL INDE DS‘ancestral | JN ONIN NE| polymorphism « | outside « of t the 1 : region a aN ees| analysed | and | | possibly ‘involved 1 {in wee pSageing ' would | | be t “eee pee: tightly | alinked I to « ) one ¢ > OF other « r of t “iveF the \ =- various | ee IE FE LI DO; haplotypes/groups « ; created { | for “Oothe | ’3*4   eS OO) region   A| this :   and « | contribute indirectly | in| abethe | SIP oeoverall ;  | analysis.

In “EN MP. addition | it —| to | ) this ; ; analysis, | the | - frequency « of | * the 1 COE NAR POE EE- mitochondrial 1 LLL LIST| nucleotide - polymorph- isms, 1 (cid:17)(cid:3)%+,: < to, (cid:14)> meeeyin. SPE ON ES| Japanese « TN FOE> centenarians sand 1 (cid:17)(cid:3);(%% <@ tO. (cid:14)> in| NEEM| French nD EEN| centenar- OOO ONDians, yaawere | “eM Xsalso ee ape) examined | “er| in renee| OUF ‘control ; | and | aged | Se ee eA| populations : in | ee A. an | it ia| attempt to. verify | the menawy Neesprevious | ; findings « of s f significant t associations ( ™ Soe; (Tanaka ¢ Let < tal., +;;:) ; Ivanova « Let < tal., +;;:> ) and to ¢  y establish \ ee Eee. whether t oethese |  - polymorphisms « a> are > also s  significantly increased | int eo =\ the | pT- Trish ¢ °F1 aged community.

  , Materials ; ; and | | methods

2*4* Subjects

were ¢ : chosen lat NE EEt random | pee \~| from t the | > DNA I _ bank « of 1 f normal | ae EES”| healthy ARNO“aucasian i AE Ieee| individuals | eM P; from the Northern Ireland population stored at the Northern BONE DOORIreland Histocompatibility and Immunogenetics “a PM peeLaboratory <(cid:13)(cid:24)9(cid:24)1> of "Belfast | city Hospital. | The isstudy ‘pee DPinvolved +’; aged « ENED| consecutive : > samples | <,(= “anna» female, ‘ 4(= » male \ ay- with ¢ | an ¢ pee| age-range |  :(A;, | years ¢ ; old) Coo_etfrom the Belfast ETElderly NE NE AMEE AINAYLongitudinal Free-living Ageing “ow eeeStudy | “ov(Rea | et. neal., ’(((>(cid:28) MeeeAll samples \ ; were | > healthy ( nog No“aucasian 1 individuals | NS; from t Oo **.the 1 ‘ono Nee> Northern | med XeIreland | | population ¢ i: display- —"*ing reeno signs of BO OIE NELLIEage-related disorders, still living NDI II NIELSindependently vNbreeesand ED I OTEapproximate ?7(cid:13)(cid:24)7D(cid:29) oN NSStatus | S SMe~w tues, (Ligthart | et cal., +;:*>(cid:28) Inv | the | Case | of | ‘the 1 (cid:17)(cid:3)%+,: we Oe Peepolymorphism : | study, DNA samples we eee”derived from “neo 2 AMEcohort of 4( unrelated Asian iS peeweeindividuals EE EEE INE YD(Singapore ~hinese, eeeage-range +:A*% years old) were accessed from the DNA eearchival SOPbank at NIHIL | 7,and ; ones| also | oOused i } in i I| order 1 to | a MENA ONS) provide | : positive : -and 1 | negative « controls | for | ‘this 1 ; particular polymorphism | -and 1 | therefore ae eeeverify | OUT | ™ pene he ®sequence NE EIN ESspecific | probe results.

2*2* PCR « amplification « Of | mtDNA | region . ) spanning | ‘nucleotides 4/(cid:29)37840<<4

mtDNA | es peegenome were |   pes Jae- essentially. those | eee eeedescribed by’ -Torroni | “MEand | ON NE NEcolleagues | <+;;’> but DERE NES ©extended ; olat t t the : %(cid:4) end t | to 1 IEDy improve t > the : » specificity « y and ~— 2s oro &| sensitivity « of t F the | / “O°OR | Seen(Table | +>(cid:28)  selected 1 | region « | of 1 F the 1 oN EEE TES» mtDNA 1 we Re Psincludes 1 ; the « BONEN JODentire | Sytb y and | ay NS| ND6 | ) genes, ;  _~ karen. small : ape an aS| section « | of the. / NDS | oegene | <’*(  ) and | | almost | half "of 1 * the | Oe DO DNhypervariable | “we oeD-loop, which | | contains | the

  s defined 1 according t   ——) the F EO> published 1 1 mitochondrial ¢ | genome s  > sequence ¢   f Anderson « et Z t al. ( <+;:+>(cid:28) cing | y primers i sin 1 Table | + ene oN| were | ee  he1 to i ) identify t the ¢ > exact I Ce ONt nucleotide \ ————> variation | causing | y presence ( ae Me> OF é yn ase ye wesr absence ¢ > of t EES OEEpolymorphic 1 > restriction : 1 sites