Logistic regression of the allele distributions of all markers confirmed the previous findings, with only the minor allele of rs275653 being significantly associated

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one of the most long-lived and includes a high number of centenarians. Data of 1000 Genomes project in Japanese population indicate that rs275653 and rs422858 are in complete LD as in Italian subjects, so we chose to geno- type rs275653 because of the availability of a commercial TaqMan assay. In accordance to 1000 Genomes and Hap- Map data, in Japanese population the minor allele of rs275653 had a lower frequency (0.084) than in Italian – controls (0.158). Case control analysis of rs275653 in Japanese cohort revealed that the GG genotype was sig- nificantly associated to longevity under the recessive and 0 0 additive models (P 0.046 and P 0.047, respectively, 0 Table 2). Genotype distribution evidenced a trend (P 0.077) while no difference was observed with the domi- nant model and at allele level (Table 2).

of the minor rs275653 G allele between Italian and Japanese cohorts, an enrichment of the GG genotype was evident in both groups of centenarians as com- pared to controls, we performed further analyses merging the two populations. In the combined analy- sis, association of rs275653 with longevity was 0 observed using the recessive (P 0.007) and the addi- 0 tive models (P 0.009) (Table 2). Also, genotypic frequencies were significantly different between cen- tenarians and controls in the overall population 0 (P 0.015). After adjusting for confounding factors, namely gender, antihypertensive treatment and popu- 0 lation of origin, logistic regression gave P 0.00034 0 and P 0.00151 for the association between longevity and rs275653 considering either genotype distribution or the recessive model, respectively (Table 3).

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FACS analysis of AT R on leukocyte populations 1

Given that the rs275653 SNP and the linked rs422858 are localized in the promoter region, we hypothesized that they might affect AGTR1 expression. We conducted in silico analyses to evaluate whether the allelic variants of rs275653 and rs422858 were associated with the presence/absence of DNA binding factors in AGTR1 promoter. Using Tess, we found that the region includ- ing the major allele variant A of rs422858 contains an E-box element (CANNTG), which can be the target of helix-loop-helix transcription factors, including Tal1 and NF-uEA. By contrast, the C allele, which is more frequent in our centenarians, causes the loss of the E-box site. No putative binding sites were predicted for allele variants of SNP rs275653 which, however, is in

Table 3 Logistic regression analysis of rs275653 and longevity in the combined group

included in the analysis Overall OR 95% CI P Model fit 2.04 1.38–3.02 0.00034 <0.00001 model 1.96 1.29–2.96 0.00151 model – nr (female vs. male) 2.76 1.94–3.91 <0.00001 treatment 3.62 2.59–5.06 <0.00001 – nr

OR odds ratio, CI confidence interval, nr not retained in the model