polyacrylamide gel'®. 2, The ACE diallelic polymorphism wasgenotyped by amplification of the variable segment and resolution ofthe 190 and 490 bp alleles on a 1.5% agarose gel'*. 3, APOBpolymorphisms were analyzed as described previously” byamplification of the two DNA segments, direct visualization of theamplified product on a 10% non-denaturing polyacrylamide gel forins/del and visualization of the Xbal digestion products on a 1.5%agarose gel for the Thr,,,, substitution. 4, The polymorphism atD4S95 was revealed by Southern hybridization with probe 674 andAccl digested samples”. 5, The BCL-2 microsatellite alleles weretyped as described by Fougerousse et al. (personal communicaton).Primers 5'-TGTGTGTGTGTAGCGCGTGT-3' and 5'-CTGGCCGTGTGAGTGTGTGT-3' were used for amplification inareaction mix containing 200 ng genomic DNA, 10 pmoles of eachprimer and 0.75 U of Taq polymerase in a final volume of 50 pl.Samples were subjected to 30 cycles consisting of 40 s at 92 °C, 30sat 62 °C and 30 s at 72 °C. The alleles were revealed by Southernhybridization with a (CA), probe after resolution of the fragmentson a 10% denaturing polyacrylamide gel.

MethodologyDec dcetee en nme

 Populations. Centenarians were recruited from all over France(regional distribution: 40% Paris region, 18% central, 4% northern,5% eastern, 15% western, 18% southern), at their home or ininstitutions, through media advertising and retirement organisations.They were all Caucasians residing in France. The criterium forinclusion in the study was to be at least 99 years old on the day ofblood collection. The population of 338 centenarians is composed of11% men and 89% women with a mean age of 100.71 + 0.13.Controls were chosen among unrelated normal individuals fromFrench CEPH pedigrees.

Acknowledgements 

———————eWe thank Fondation IPSEN, ARRCO, AGIRC and the participatingclinicians as well as E. Sénard and J. Boirot for help in collecting thecentenarians, F. Lethrosne, F. Dufour, C. Habita, M. Legrand, L.Gressin, C. Massart, C. Billon for technical assistance, M.R. Hayden forprobe 674; F. Cambien, T. Kirkwood and R. Moulias for support anddiscussion.

Genotypes. 1, The APOE genotypes were analyzed by Hhal digestionofa244 bp PCR- amplified fragmentcontaining the two polymorphicsites, with the restriction fragments resolved ona 10% non denaturing

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