Note: OR ¼ odds ratio; CI ¼ confidence interval.

Table 3. Angiotensin-Converting Enzyme (ACE) Genotypes in Relation to Sex and Age in 1344 Healthy Elderly Persons and 64 Centenarians

Men (N ¼ 592) Women (N ¼752) All (N ¼ 1344) Age Range Group (N) (Mean Age 6 SD) D/ p OR (95% CI) I/ D/ p OR (95% CI) I/ D/ p OR (95% CI) I/ All 22–90 86.82% — — 56.59% 87.50% — — 57.18% 87.20% — — 56.92% (n ¼ 1344) (41.31 6 13.63) (n ¼ 514) (n ¼ 335) (n ¼ 658) (n ¼ 430) (n ¼ 1172) (n ¼ 765) Group 1 Age   30 86.54% .420 1.548 54.81% 86.70% .926 1.040 62.77% 86.84% .498 1.233 59.93% (n ¼ 292) (27.29 6 2.10) (n ¼ 90) (0.581–3.664) (n ¼ 57) (n ¼ 163) (0.459–2.353) (n ¼ 118) (n ¼ 253) (0.672–2.262) (n ¼ 175) Group 2 30 , age , 45 85.91% .288 1.538 55.37% 87.92% .858 0.932 54.68% 86.96% .521 1.199 55.01% (n ¼ 629) (36.76 6 3.94) (n ¼ 256) (0.692–3.419) (n ¼ 165) (n ¼ 291) (0.430–2.020) (n ¼ 181) (n ¼ 547) (0.688–2.089) (n ¼ 346) Group 3 45 - ~~SIA age , 60 86.92% .462 1.411 62.62% 87.73% .901 0.948 57.06% 87.41% .653 1.153 59.26% (n ¼ 270) (50.44 6 4.17) (n ¼ 93) (0.562–3.543) (n ¼ 67) (n ¼ 143) (0.408–2.200) (n ¼ 93) (n ¼ 236) (0.620–2.141) (n ¼ 160) Groups 1–3 Age , 60 86.25% .304 1.495 56.78% 87.54% .924 0.965 57.48% 86.99% .507 1.197 57.18% (n ¼ 1191) (37.54 6 8.78) (n ¼ 439) (0.691–3.233) (n ¼ 289) (n ¼ 597) (0.462–2.014) (n ¼ 392) (n ¼ 1036) (0.703–2.037) (n ¼ 681) Group 4 60 —VIA age  90 90.36% Reference 55.42% 87.14% Reference 54.29% 88.89% Reference 54.90% (n ¼ 153) (70.71 6 7.51) (n ¼ 75) (n ¼ 46) (n ¼ 61) (n ¼ 38) (n ¼ 136) (n ¼ 84) Centenarians 98   age   108 83.33% .386 0.533 61.11% 93.48% .273 2.115 50.00% 90.63% .705 1.208 53.13% (n ¼ 64) (100.28 6 1.86) (n ¼ 15) 0.127–2.247 (n ¼ 11) (n ¼ 43) (0.541–8.270) (n ¼ 23) (n ¼ 58) (0.453–3.220) (n ¼ 34)

Note: SD ¼ standard deviation; OR ¼ odds ratio; CI ¼ confidence interval.

(sex-adjusted OR values ¼ 1.581; 95% CI, 1.211–2.064 and 1.357; 95% CI, 1.048–1.759, respectively, for I and D alleles), Group 2 (sex-adjusted OR values ¼ 0.753; 95% CI, 0.600–0.945 and 0.775; 95% CI, 0.628–0.956, respectively, for I and D alleles), and Group 4 (sex-adjusted OR values ¼ 0.669; 95% CI, 0.455–0.983 and 0.637; 95% CI, 0.453– 0.897, respectively, for I and D alleles). No relationship was found between apoE and ACE alleles in the different age groups, even after adjustment for sex (Table 5).

significantly from the expected Hardy–Weinberg frequen- cies (p . .999): Expected allele frequencies were I ¼ 0.313 and D ¼ 0.688.

3), no significant differences were observed in the frequency of D participants in the oldest age group compared to participants less than 60 years of age, that is, Groups 1–3 (88.89% vs 86.99%, p¼.507), Group 3 (88.89% vs 87.41%, p ¼ .653), and Group 2 (88.89% vs 86.96%, p ¼ .521). Furthermore, no difference was found in the frequency of D participants in the oldest age group compared to centenar- ians (88.89% vs 90.63%, p ¼ .705). In a linear model, no

D ISCUSSION

This study investigated the role of sex in apoE and ACE polymorphism and their association with healthy aging and longevity. We studied two genetically homogeneous groups of Caucasians participants who lived in southern Italy, that is, a cohort of healthy participants and a group of cente- narians. In these populations, we found a significantly higher prevalence of the e2 allele in participants older than 60 years and in centenarians than in younger participants. These findings suggest a role of the e2 allele in healthy aging and longevity, and confirm previous studies reporting a significant increase in the e2 allele frequency with advancing age, including centenarians (26–28).

In a sex 3 genotype analysis, no significant differences were found in the 1344 participants (p ¼ .875) or in cente- narians (p ¼ .414). As expected, after dividing participants according to sex, no differences in the distribution of ACE allele among the age groups were found. Logistic regression analysis (Table 4) confirmed the lack of a significant association between the D allele and age. After adjustment for sex, however, a significant association between the D allele and age was observed in the oldest age group (crude OR ¼ 1.177; 95% CI, 0.693–2.001; sex-adjusted OR ¼ 0.453; 95% CI, 0.453–0.897) and centenarians (crude OR ¼ 1.419; 95% CI, 0.603–3.338; sex-adjusted OR¼2.135; 95% CI, 2.135–3.636).

An association of the e2 allele with aging may be ex- plained by two hypotheses. First, the progressively in- creased frequency of the e2 allele with aging may have been a consequence of the disappearance in our selected healthy participants of the e4 allele, which is reported to be linked to a higher risk of age-related diseases (9) and mortality (29).

Notably, heterosis was observed when ACE genotypes were classified according to sex. In fact, a significant association of both I and D alleles was shown in Group 1

Table 4. Association of Angiotensin-Converting Enzyme (ACE) Alleles With Age in 1344 Healthy Elderly Persons and 64 Centenarians

I D Crude Values Adjusted Values Crude Values Adjusted Values Group (N) p OR (95% CI) p OR (95% CI) p Crude OR (95% CI) p Adjusted R (95% Group 1 (n ¼ 292) .218 1.179 (0.907–1.533) .001 1.581 (1.211–2.064) .680 0.923 (0.630–1.351) .021 1.357 (1.048–1.759) Group 2 (n ¼ 629) .237 0.880 (0.712–1.088) .014 0.753 (0.600–0.945) .689 0.938 (0.684–1.285) .018 0.775 (0.628–0.956) Group 3 (n ¼ 270) .354 1.136 (0.867–1.487) .375 1.135 (0.858–1.503) .978 1.006 (0.674–1.500) .246 1.170 (0.897–1.527) Group 4 (n ¼ 153) .626 0.919 (0.656–1.288) .041 0.669 (0.455–0.983) .547 1.177 (0.693–2.001) .010 0.637 (0.453–0.897) Centenarians (n ¼ 64) .550 0.858 (0.519–1.418) .510 1.192 (0.707–2.012) .423 1.419 (0.603–3.338) .005 2.135 (1.253–3.636)

Notes: OR ¼ odds ratio; CI ¼ confidence interval.

5. Shared Frequencies of Apolipoprotein E (ApoE) and Angiotensin-Converting Enzyme (ACE) Alleles in a Cohort of 1344 Healthy Elderly Persons and 64 Centenarians