Alleles 694V (%) 694I (%) 726A (%) AMI patients 30 (12%) 0 (0%) 0 (0%) Young controls 25 (6%) 0 (0%) 0 (0%) Centenarians 5 (4%) 0 (0%) 0 (0%)

Significance was obtained by (cid:3)2 (3(cid:4)2 table) for M694V alleles, as the alleles were distributed unevenly among the three cohorts (P(cid:6)0.0033).

TABLE 3. Prevalence of the Heterozygous M694V Genotype in 121 AMI Patients, 196 Controls, and 68 Centenarians from Sicily

in early atherogenesis. Moreover, adhesion molecules on the leukocyte membrane are increased in patients with AMI with respect to control subjects, causing the recruitment of neutro- phils and monocytes to the involved myocardial region [28– 30].

Genotypes MM (%) MV (%) AMI patients 91 (75%) 30 (25%) Young controls 171 (87%) 25 (13%) Centenarians 63 (93%) 5 (7%)

The aim of this report was to investigate whether the proin- flammatory mutations of the MEFV gene play an opposite role in AMI and longevity. To this purpose, all the subjects were screened for M694V, V726A, and M694I, which are the most frequent FMF-associated mutations. Also, a fourth mutation, E148Q, is quite frequent in the Mediterranean area, but its penetrance is low [19, 20]. We found that only the M694V mutation, which is the most frequent in all Mediterranean populations, was present in all the groups studied [19, 20]. This proinflammatory mutation was over-represented signifi- cantly in patients with AMI and under-represented in cente- narians. The differences between patients and age-related con- trols are significant but relatively small, as demonstrated by OR values. However, as AMI is a multifactorial disease, any single mutation will only provide a small or modest contribu- tion to risk, also depending on interaction with a particular environment [31].

Significance was obtained by (cid:3)2 (3(cid:4)2 table) for the heterozygous genotype, which was distributed unevenly among the three cohorts (P(cid:6)0.0019).

was present in all three cohorts of study. It was most frequent in AMI patients (12%), intermediate in healthy, young controls (6%), and less frequent in centenarians (4%; Table 2, P(cid:6) 0.0033). All the genotypes were in HWE. As expected from the HWE of M694V genotypes, all subjects with the M694V mutation were heterozygous. The heterozygous genotype was also significantly overexpressed in AMI patients (30/121) and underexpressed in centenarians (5/68) compared with young controls (121/196; Table 3, P(cid:6) 0.0019).

By performing a series of 2 2 separate comparisons, high significance for the different frequencies of M694V genotypes and alleles was obtained between AMI patients and age- matched and centenarian controls. The frequency of M694V proinflammatory genotype and allele in centenarians was lower (but not significantly) than that observed in young, healthy controls. Finally, we performed a logistic regression analysis to test the association of genotypes with AMI after adjustment for smoking habits, family history of CHD, and the presence of Type 2 diabetes, obesity, hypertension, hypercholesterolemia, and hypertriglyceridemia. As a result of the different number of women in the groups under study, logistic regression was performed only in men. Taking into account the risk factors, significant differences in genotype frequency persisted be- tween male AMI patients and male young controls and between male AMI patients and male centenarians (Table 4). These results indicate that the M694V genotype is an independent risk factor for developing AMI in the Sicilian population.

From literature, several case-control studies show that MEFV mutations do not seem to predispose toward inflamma- tory diseases such as Crohn’s disease, multiple sclerosis, or rheumatoid arthritis, but patients having that inflammatory burden and at the same time, carrying one MEFV mutation appear to be highly susceptible to develop a more severe disease [17, 32, 33]. So, it may be possible that MEFV muta- tions are not increasing the susceptibility to develop athero- sclerosis, but they might affect its severity and complications, enhancing AMI occurrence.

Taking into account that several mechanisms have been proposed to explain the anti-inflammatory role of pyrin [7, 8, 34, 35], it may be intriguing to suggest a mechanism by which the mutated MEFV gene could be responsible for this process. One option is that the mutated pyrin allele is not able to limit the migration of neutrophils and activated monocytes into the site of inflammation, and although failing to down-regulate the inflammatory response of monocytes, instead augments the secretion of inflammatory molecules. Several findings are con- sistent with this role. First, the inflammatory attacks of FMF are prevented with colchicine, an agent that disrupts microtu- bules by blocking polymerization of tubulin [11]. Second, the wild-type pyrin is considered anti-inflammatory, as by induc- ing apoptosis of neutrophils and monocytes, it down-regulates the recruitment toward the inflammatory sites [7]. Moreover, recent studies have demonstrated that white blood cell (WBC)

DISCUSSION

The role of inflammatory responses in mediating all stages of atherosclerosis and CHD has been well established. Athero- sclerosis begins as a focal process at specific regions of the vasculature, so-called lesion-prone areas, where hemodynamic flow is altered. The arterial endothelium expresses numerous adhesion molecules. The specific up-regulation of adhesion molecules suggests that they may regulate leukocyte adhesion

 TABLE 4. 2 2 Comparisons between the Different Groups with OR and CI

Young controls Centenarians   AMI patients (M694V genotypes) P 0.009 OR 2.2 (1.3–4.1) P 0.003 OR 4.2 (1.5–11.3)   AMI patients (M694V alleles) P 0.009 OR 2.1 (1.2–3.6) P 0.004 OR 3.7 (1.4–9.8)