PROCESSED TEXT:

0.309 0.219 Constant 
0.346 0.328 0.291 Diplotype A-C/G-A 
Age 0.014 0.006 0.014 0.026 to 0.003 [Sex=males] 
0.376 0.190 0.048 0.003 0.748 Constant 
0.903 0.452 0.045 Diplotype A-C/G-C 
Age 0.001 0.005 0.892 0.009 
[Sex=males] 0.166 0.161 0.300 0.481 0.148 Constant 
1.227 0.389 0.002 Diplotype G-A/G-A 
Age 0.010 0.019 0.614 0.048 
[Sex=males] 0.343 0.650 0.599 1.617 0.932 Constant 
3.615 1.505 0.016 Diplotype G-A/G-C 
Age 0.007 0.013 0.585 0.032 
[Sex=males] 0.194 0.427 0.649 1.031 0.643 Constant 
3.
035
1.
009
		3.
051
1.
076
Diplotype G-C/G-C Age 
		activity.
Each transfection experiment was carried out three times in duplicate.
Statistical analyses were performed using R statistical software for Windows (www.cran.com).
One-way analysis of variance (ANOVA) and Student's t-test with a significance level of 0.05 were adopted to check the significance of the difference between the fold induction values of the constructs containing the different haplotypes at basal conditions and after hormone stimulation.
Fig. 2 shows the observed and estimated frequency for diplotype A-C/A-C (A) and A (B) carriers.
harmful haplotype.
The slope parameters for both A-C and G-C haplotypes were not statistically different from 0, suggesting that their frequencies were independent of age.
Fig. 1 shows the frequency patterns for the G-A haplotype carriers.
As expected, both the observed and fitted frequencies indicated that there was a constant decline of this haplotype with increasing age.
In order to check if the detrimental effect of the G-A haplotype observed in both sexes involved a particular diplotype, we used the same model to analyze the frequency of the carriers of each of the six reconstructed diplotypes as a function of age.
Table 4 reports the parameter estimates for all the reconstructed diplotypes.
Diplotype A-C/A-C showed a significant positive slope parameter (=0.009; p=0.032), indicating that the frequency of subjects with this diplotype increases with age.
The diplotype A-C/G-A also showed a significant negative slope parameter (=0.014; p=0.014), which indicates that the frequency of subjects with this diplotype decreases with age (Fig. 2).
As for the haplotypes, if one considers an age interval of 15 years, the corresponding ORs from the relevant fitted models were 1.144 and 0.811 for diplotype A-C/A-C and A-C/G-A, respectively.
The diplotype A-C/G-C did not show any significant variation with age.
The other diplotypes failed to reach the level of significance probably due to their low frequencies.
On the whole, these results indicate that the A-C haplotype has a recessive and beneficial effect, while the G-A haplotype has a dominant and deleterious effect on human aging and longevity.

3. Results

3.1. Haplotype/diplotype analyses

On a sample of 910 subjects categorized by age and sex as reported in Passarino et al. (2006).
In each group, there was no evidence of departure from Hardy-Weinberg equilibrium (p > 0.05).
The two SNPs, which are very close to each other, resulted in linkage disequilibrium (D=0.99, r2=0.58).
Frequency distributions of two-marker haplotypes and diplotypes are summarized in Tables 1 and 2, respectively.
We modeled the frequency of the carriers of each of the reconstructed haplotype/diplotype as a function of age by using the binary logistic regression model.
Table 3 reports the maximum likelihood estimations of the parameters of the fitted haplotype-basedindividuals: effects of cold exposure and adiposity. Diabetes  , 

The A-C and G-A haplotypes have a positive effect on UCP1 transcriptional activity compared to the GC haplotype. However, they have opposite effects on longevity, which may be explained by considering the haplotype-specific effect after hormonal stimuli. Progesterone up-regulates while estradiol down-regulates the transcriptional activity of the AC haplotype, while neither changes the transcriptional activity of GA. Therefore, it is likely that the real in vivo effect of these haplotypes on complex phenotypes such as aging and longevity is mediated by the intricacy of different cellular and physiological stimuli. Similarly, this may explain why the G-C haplotype, lacking any transcriptional activity, did not show significant changes in its frequency during aging. This observation calls for a better understanding of the different stimuli affecting UCP1 transcription.

Human aging is accompanied by changes in energy balance and body composition which are consequent to hormonal changes (Hughes et al., 2004). In this context, the role of BAT is particularly important since it regulates energy expenditure in the elderly. Although the amount of BAT in human adults is low, it is responsible for 12% of energy expenditure, which is enough to prevent weight gain of about 2 kg per year (Lean, 1989). Consistently, an inverse relation between BAT and BMI, body fat, and visceral fat is observed in aging (Mattson, 2010; Vijgen et al., 2011). UCP1, which is expressed in BAT, has been observed to influence metabolism and fat presence of consensus sex steroid responsive elements within the UCP1 sequence is very intriguing. Indeed, there are several reports considering sex steroid hormones as modulators of the thermogenic capacity and activity of BAT (Monjo et al., 2003; Valle et al., 2008). The different effects of the UCP1 haplotypes could be explained by considering changes in the regulation of metabolism with aging, possibly mediated by age-related variations in hormonal status. Moreover, as suggested by Fessele et al. (2002), the importance of the sequence context in determining the functionality of a regulatory region clearly emerges. In fact, although the two polymorphisms did not alter either the binding site for progesterone or estradiol, a clear haplotype-specific effect on transcriptional activity was observed following hormonal stimulation.