According to the observed LD pattern, were independently inferred from 50 to

Table 1 Genotype and allele distribution

Markers rs Genotypes Controls Octogenarians code (alias) alleles All subjects Males Females All subjects Males Females n (%) n (%) n (%) n (%) n (%) n (%) rs11568820 (Cdx) GG 61 (59.2) 21 (60) 40 (58.8) 63 (69.2) 18 (69.2) 45 (69.2) GA 38 (36.9) 12 (34.3) 26 (38.2) 25 (27.5) 6 (23.1) 19 (29.3) AA 4 (3.9) 2 (5.7) 2 (3) 3 (3.3) 2 (7.7) 1 (1.5) G 160 (77.7) 54 (77.1) 54 (77.1) 151 (83) 42 (80.8) 109 (83.8) A 46 (22.3) 16 (22.9) 16 (22.9) 31 (17.0) 10 (19.2) 21 (16.2) Genotype failurea 7 (6.4) 0 (0) 7 (9.3) 13 (12.5) 6 (18.8) 7 (9.7) rs4516035 (A1012G) GG 41 (37.6) 14 (40) 27 (36.5) 29 (29.3) 9 (30) 20 (29) GA 56 (51.4) 18 (51.4) 38 (51.4) 55 (55.5) 15 (50) 40 (58) AA 12 (11) 3 (8.6) 9 (12.1) 15 (15.2) 6 (20) 9 (13) A 138 (63.3) 46 (65.7) 46 (65.7) 113 (57.1) 33 (55) 80 (58) G 80 (36.7) 24 (34.3) 24 (34.3) 85 (42.9) 27 (45) 58 (42) Genotype failure 1 (0.9) 0 (0) 1 (1.3) 5 (4.8) 3 (9.4) 2 (2.8) rs2228570 (FokI) CC 43 (39.1) 13 (37.1) 30 (40) 46 (46.5) 15 (46.9) 31 (46.3) CT 58 (52.7) 18 (51.4) 40 (53.3) 39 (39.4) 10 (31.2) 29 (43.3) TT 9 (8.2) 4 (11.5) 5 (6.7) 14 (14.1) 7 (21.9) 7 (10.4) C 144 (65.5) 44 (62.9) 44 (62.9) 131 (66.2) 40 (62.5) 91 (67.9) T 76 (34.5) 26 (37.1) 26 (37.1) 67 (33.8) 24 (37.5) 43 (32.1) Genotype failure 0 (0) 0 (0) 0 (0) 5 (4.8) 0 (0) 5 (6.9) rs1544410 (BsmI) GG 37 (33.6) 10 (28.6) 27 (36) 30 (29.2) 10 (31.2) 20 (28.2) GA 50 (45.5) 17 (48.5) 33 (44) 56 (54.3) 20 (62.5) 36 (50.7) AA 23 (20.9) 8 (22.9) 15 (20) 17 (16.5) 2 (6.3) 15 (21.1) G 124 (56.4) 37 (52.9) 37 (52.9) 116 (56.3) 40 (62.5) 76 (53.5) A 96 (43.6) 33 (47.1) 33 (47.1) 90 (43.7) 24 (37.5) 66 (46.5) Genotype failure 0 (0) 0 (0) 0 (0) 1 (1.0) 0 (0) 1 (1.4) rs17878969 (PolyA) LL 43 (39.8) 10 (28.6) 33 (45.2) 36 (35.3) 11 (34.4) 25 (35.7) LS 48 (44.5) 17 (48.6) 31 (42.5) 44 (43.1) 12 (37.5) 32 (45.7) SS 17 (15.7) 8 (22.8) 9 (12.3) 22 (21.6) 9 (28.1) 13 (18.6) L 134 (62) 37 (52.9) 37 (52.9) 116 (56.9) 34 (53.1) 82 (58.6) S 82 (38) 33 (47.1) 33 (47.1) 88 (43.1) 30 (46.9) 58 (41.4) Genotype failure 2 (1.8) 0 (0) 2 (2.7) 2 (1.9) 0 (0) 2 (2.8) Global genotype failure 10 (1.8) 0 (0) 10 (2.6) 26 (5.0) 9 (5.6) 17 (4.7)

Number of subjects with missing genotypes

Prevalence (all subjects) >g60%50%40%30%20%10%PHASE PAC-likelinood P=0.01GLAS ALHaplotypes at 3' region  GSPrevalence (males)PHASE PAC-likelinood P=0.0170%60%50%40% neck30% P=0,00620%10%0%GL AS AL 6sHaplotypes at 3' region

Prevalence (males)PHASE PAC-likelinood P=0.0170%60%50%40% neck30% P=0,00620%10%0%GL AS AL 6sHaplotypes at 3' region

implemented in PHASE 2.1.1 (Li and Stephens 2003). Haplotype-specific test was performed by 2 9 2 contingency table analysis. Experiment-wide significance threshold required to keep Type I error rate at 5% was P = 0.036

Fig. 2 30UTR haplotype prevalence in controls (CTR) and octogenarians (OCT). Histograms plot of the 30UTR haplotype prevalence in all subjects (a) and in males (b). Bars indicate 95% confidence interval of the proportion. Global distribution of haplotypes was compared by the PAC-likelihood test

were mainly due to males (0.07% in controls versus 9.5% in octogenarians, OR out of range; P = 0.014; Fig. 3b). The distribution of carriers of at least one copy of GGCGS haplotype (0.5% in controls and 8.3% in octogenarians, P = 0.023) is presented in Table 2. Males carrying at least one copy of GGCGS haplotype represent just 0.1% in controls but 18.9% in octoge- narians (P = 0.012).

(PHASE PAC-likelihood test P = 0.01) (Fig. 2b). Haplotype-specific test revealed that rs1544410- G:rs17878969-S (GS) haplotype was overrepresented in octogenarians (7.3 versus 0.8%; odds ratio (OR): 15.3, 95% confidence interval (CI): 2.3–637; P \ 0.001) (Fig. 2a). This difference was greater in males (14.2% in octogenarians versus 0.3% in controls, OR out of range, P = 0.006) (Fig. 2b). We also estimated the haplotype carrier status of each individual. The distribution of carriers of at least one copy of 30UTR GS haplotype is presented in Table 2. Carriers of at least one copy of GS haplotype were common in octogenarians (13.3%) but rare in controls (1.6%) (P = 0.008). Among males, nearly a quarter of the octogenarians carried at least one copy of GS haplotype, whereas GS carriers represent less than 1% of controls (24.4 versus 0.7%, respec- tively; P = 0.013).

Discussion

Long-lived people escaped or delayed major age- related illnesses such as cardiovascular diseases and cancer, as well as life-threatening infections and immune-related diseases. It is noteworthy that vita- min-D plays a key role in these impairing diseases (Holick 2005). A general concern exists about the vitamin-D insufficiency that has been found in older persons (Holick 2006). Recent data indicates that low serum levels of 25-hydroxyvitamin-D are associated with all-cause mortality in the general population (Dobnig et al. 2008) as well as in older men and women (Pilz et al. 2009). However, the question of whether vitamin-D deficiency is a cause or a conse- quence of poor health status remains to be answered.

Extended five-marker haplotypes were also esti- mated. Prevalence in controls and octogenarians is presented in Fig. 3. Extended five-marker haplotype counts and frequencies were compared between groups. Global five-marker haplotype distribution showed statistically significant differences (PHASE PAC-likelihood test P = 0.020) (Fig. 3a), again mainly contributed by differences in males (PHASE PAC-likelihood test P = 0.020) (Fig. 3b).

in the aging process (Lanske and Razzaque 2007; Tuohimaa 2009). Studies in animal models have revealed a paradoxical effect of vitamin-D on aging. Genetically modified mice lacking FGF23 and Klotho genes exhibit features of premature aging that have been related with an altered mineral homeostasis and