Five-marker haplotype-specific test revealed that rs11568820-G:rs4516035-G:rs10735810-C:rs15444 10-G:rs17878969-S (GGCGS) haplotype was differ- entially distributed between groups (0.25% in controls versus 4.1% in octogenarians, OR: 11.5; 95% CI: 1.2– 621, P = 0.024; Fig. 3a). Again, these differences

Table 2 Distribution of carriers of at least one copy of selected haplotypes in controls and octogenarians

Carriersa for: Controls countsb (%) Octogenarians countsb (%) All subjects Males Females All subjects Males (n = 101) (n = 35) (n = 66) (n = 86) (n = 25) (n = 30UTR markers 1.6 (1.6) 0.2 (0.7) 1.4 (2.1) 11.4 (13.3) 6.1 (24.4) 5.3 haplotype GSc Five-marker haplotype 0.5 (0.5) 0.05 (0.1) 0.45 (0.7) 7.1 (8.3) 4.7 (18.9) 2.4 GGCGSd

Note: As the number of each haplotype carrier was statistically inferred values obtained for counts were in fractional numbers a Carriers were those bearing one or two copies of the corresponding haplotype b Counts refer to the number of haplotype carriers that were statistically inferred from PHASE 2.1.1 ‘‘out_pairs’’ output file c Haplotype rs1544410-G:rs17878969-S. Fisher exact test, octogenarians versus controls: all subjects, odds ratio (OR): 7.1; 95% confidence interval (CI): 1.5–66.3, P = 0.008; males, OR out of range, P = 0.013. Experiment-wide significance threshold required to keep Type I error rate at 5% was P = 0.036 d Haplotype rs11568820-G:rs4516035-G:rs10735810-C:rs1544410-G:rs17878969-S. Fisher exact test, octogenarians versus controls: all subjects, OR: 11.9; 95% CI: 1.3–650, P = 0.023; males, OR out of range, P = 0.012. Experiment-wide significance threshold required to keep Type I error rate at 5% was P = 0.011

d Haplotype rs11568820-G:rs4516035-G:rs10735810-C:rs1544410-G:rs17878969-S. Fisher exact test, octogenarians versus controls: all subjects, OR: 11.9; 95% CI: 1.3–650, P = 0.023; males, OR out of range, P = 0.012. Experiment-wide significance threshold required to keep Type I error rate at 5% was P = 0.011

sOLoVs929V‘SOlvysoovWLoVvwoovsolvs9199svoov‘SvLOWs92v91919¥WwiveWoy$999919990Wis.Wwo99WivoWwovo‘SvlvoSvLvV‘Div‘SVLODsvvV19199Tovsv299WDLvdsv19999WIVD ctr Ooct        Haplotypespecific testP=0.024=0.02  PHASE PAC-likelihood P:Rk Rk KC KR KKS84 SH SHSma AA{(s}Da/qns |je) aduajenaigExtended haplotypessoLovs909V‘SOLsoovwiovwoovsalvos9L99svooV‘SVLOVs9ov9WD19vWiWwovv9999IOVWwio9woooWwivoWwovoSvlvDSVLVlwSVLODSvovV19199199vVsv299Divosv‘19999‘92v9       =0.014Haplotypespecific testP:PHASE PAC-likelihood P=0.02$5 8%eRe &x(sayew) aduajenaigExtended haplotypes

Fig. 3 Five-marker haplotype prevalence in controls (CTR) and octogenarians (OCT). Histogram plot of five- marker 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 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 = 0.011

hormonal forms of vitamin-D where both hyper- and hypovitaminosis are risk factors for abnormal aging (Tuohimaa 2009). This bimodal effect of vitamin-D levels also has been described for vascular disease risk in children on dialysis (Shroff et al. 2008) and prostate cancer risk in Nordic countries (Tuohimaa et al. 2004). Therefore, it can be assumed that for good

hypervitaminosis-D due to increased 1-a-hydroxilase activity (Razzaque and Lanske 2006). At the same time, null mice for the vitamin-D receptor gene, in which vitamin-D action cannot occur, also show features related with premature aging (Keisala et al. 2009). According to this, it has been proposed that aging shows a hormetic U-shaped dependency on

the common 30UTR haplotypes GL and AS been linked to low and high mRNA stability, (Fang et al. 2005). However, no causa- effect has been assigned to rs1544410 or polymorphisms. The main resulting is to consider those polymorphisms as for an unknown causative element located in 30UTR region. In octogenarians, we found an of the GS haplotype prevalence. GS haplo- combines elements of the two common GL and haplotypes, from which we can expect an phenotype. Assuming the vitamin-D on the aging process follows a hormetic model (Tuohimaa 2009), haplotypes con- low (GL) and high (AS) VDR mRNA stability to an adverse vitamin-D response caused deficiency and toxicity, respectively. However, conferring a middle VDR mRNA stability GS) can be considered as protective, as vitamin- response can remain in the homeostatic region,

is responsible for its genomic action. Several VDR gene polymorphic variants have been described as affecting VDR gene expression (rs11568820 and rs4516035), protein structure (rs10735810) and mRNA stability (rs1544410 and rs17878969) (Fang et al. 2005; Uitterlinden et al. 2004a). Variability at the VDR locus has been associated with cancer, osteoporosis, cardiovascular diseases and diabetes (Uitterlinden et al. 2004b) as well as mycobacterium (Bellamy and Hill 1998) and viral infections (Fibla and Caruz 2010).