No effect of the polymorphism on aggregate CVD phenotype was found (Fig. 1D). Nevertheless, a significant effect was observed for more homogeneous phenotype of myocardial infarction (MI) (Fig. 1E) that is in line with prior studies (Heckbert et al., 2003). We also hypothesized that MI could share common genetic background with intermittent claudication (IC) because both conditions could be linked to the development of systemic atherosclerosis. Indeed, IC is a symptom of peripheral arterial disease (PAD) and the latter is often a sign of widespread accumulation of fatty deposits in arteries, i.e., atherosclerosis. This condition may reduce blood flow not only to legs but also to heart and brain. Atherosclerosis may thus be a common risk factor for MI and PAD/IC (Murabito et al., 2002). As expected, the detrimental role of the Gln27Gln homozygote attains better significance for the compound phenotype of MI and IC, especially in men (Fig. 1F).

3.3. Genetic effects on longevity

Our analyses did not reveal significant association of the Gln27Glu polymorphism with mortality risk. For both sexes combined the RRs were RR = 1.20 (CI = 0.72–2.00) for the Gln27Gln homozygote and RR = 1.25 (CI = 0.78–2.03) for the Gln27Glu heterozygote compared to the Glu27Glu homozygote.

Because the Gln27Glu polymorphism is associated with risks of cancers and MI, and because these diseases are among the leading causes of death in developed societies, this polymorphism might

Logarithm of Odds Ratios(A) Gin?’Glu              72 74 76 78 80Cut-off for longevity, years    

Probability of staying free of diseases0.960.920.880.840.80.950.90.85        M&W; CancerN=1489 (252)—— Gly*GlyiGiu?Glu; Refae Arg'*GlyiGin27GiuRR=1.75"; Cl=1.13-2.70_._ Gly8GiyiGineGiuL RR=1.83"; Cl=1.15-2.91Arg'sGly /GIn??GInPr RR=2.11"; Cl=1.32-3.38oe Arg'sArg /GIn?7GIn[- RR=2.14"; Cl=1.32-3.48_ _ _ Gly'®Gly /GIn?7GInRR=2.20°, CI=1.12-4.34 ‘(A)      Men; MI&ICN=754 (103)—-— Gly'8Gly/Gin??Glu; Refa Arg'sGlyiGin?7Glufs RR=1.04; Cl=0.56-1.93Gly*GiyiGlu?GluRR=1.06; Cl=0.53-2.12[____ Arg'*Arg /GIn2"GInRR=1.33; Cl=0.67-2.63_ _ Gly'8Gly /GIn?’Ginf= RR=1.45; Cl=0.53-3.96Arg*8Gly /GIn?7GIn(B)_RR=1.95", Cl=1.04-3.660 100 200 300 400Time, months 

Fig. 3. Patterns of chances to live long life for carriers of the (A) Gln27Glu and (B) Glu27Glu genotypes and (C) Glu27 allele compared to the Gln27Gln genotype (horizontal line). The x-axis shows varying cut-off for longevity (see Section 2.2). Thelogarithmofoddsratiosisshownformodelswithadjustmentfor(i)ageandsex (crosses); (ii) age, sex, systolic and diastolic blood pressures, total and high-density lipoprotein cholesterol, body-mass index, smoking, and diabetes (filled dots), and (iii) life span without cancer (solid line). Bars show 95% confidence interval.

analysis) decreases because living individuals who are of cut-off ages or younger have to be censored out due to unknown life span. This results in a sample of N = 135 SL subjects and N = 30 LL subjects at cut-off = 86 years. These constraints limit flexibility of the cut-off ages.

Fig. 2. Probability of staying free of (A) cancer and (B) compound phenotype of myocardial infarction (MI) and intermittent claudication (IC) for (A) men and women and (B) men carrying compound genotypes of the ADRB2 Arg16Gly and Gln27Glu polymorphisms (patterns of probability are ordered according to increasing relative risks; see insets). Protective Gly16Glu27 haplotype is italicized andunderlined(giventhattheArg16Glu27haplotypeisvirtuallylacking,seeTable1, haplotypes are uniquely determined from the respective genotypes). The letter ‘‘N’’ intheinsetdenotessamplesizewithnon-missinginformationonallcovariates.The numbers in parentheses are counts of individuals with onsets of respective condition within the follow-up period (months). The RR (CI) denotes relative risks (confidence intervals) calculated using the Cox regression model adjusted for age, sex (whennecessary), systolic and diastolic blood pressures, totaland high-density lipoprotein cholesterol, body-mass index, smoking, and diabetes. *0.01 < p   0.05; **p D<C 0.01.

Carriers of the Gln27Glu and Glu27Glu genotypes and the Glu27 allele tend to systematically live shorter lives when longevity is defined as reaching young-old ages (e.g., cut-off = 72 years) and longer lives when longevity is defined as reaching oldest-old ages (e.g., cut-off = 83 years) compared to carriers of the Gln27Gln genotype. These patterns are consistent and essentially the same for overall longevity and healthy life span (solid line in Fig. 3 shows representative pattern for longevity without cancer). They also remain the same irrespective of adjustment for health-related risk factors. Estimates of the associations of compound genotypes with longevity are not reliable due to small sample sizes.

4. Discussion and conclusions

be involved in characterization of overall and healthy longevity (see definition of the longevity phenotypes in Section 2.2). Given that incidence rate of these diseases begin to notably increase in adulthood (at about 50 years), cut-off for longevity was left flexible conditional on the available sample size (see next paragraph). This technique helps also offset the chance nature of findings provided that the revealed associations exhibit consistent (but not stochastic) pattern across age.

In this work, we investigated associations of genetic variants of two common polymorphisms of the ADRB2 gene, Arg16Gly and Gln27Glu, with the risks of cancer, CVDs, mortality, and longevity. Our analyses show that the Gln27Glu polymorphism, but not the Arg16Gly polymorphism is associated with selected aging-associ- ated phenotypes. The analyses suggest two insights on: (i) the systemic role of the ADRB2 gene in healthy aging and (ii) mechanisms linking systemic genetic health effects with overall and healthy longevity in evolutionary context.