The text appears to be a scientific article discussing the association of Apolipoprotein E and Angiotensin-Converting Enzyme gene polymorphisms with healthy aging and longevity. Here is the processed text:

Title: Differences in the Association of Apolipoprotein E and Angiotensin-Converting Enzyme Gene Polymorphisms With Healthy Aging and Longevity: A Population-Based Study From Southern Italy

Authors: Davide Seripa, Marilisa Franceschi, Maria G. Matera, Francesco Panza, Patrick G. Kehoe, Carolina Gravina, Giuseppe Orsitto, Vincenzo Solfrizzi, Giovanni Di Minno, Bruno Dallapiccola, and Alberto Pilotto

Affiliations: 
1. Laboratory of Geriatrics and Gerontology, Research Department
2. Geriatric Unit, Medical Sciences Department, IRCCS
3. Department of Geriatrics, Center for Aging Brain, Memory Unit, University of Bari, Italy.
4. Care of the Elderly, Department of Clinical Science at North Bristol, University of Bristol, United Kingdom.
5. Laboratory of Hemostasis and Thrombosis, Research Department, IRCCS Casa Sollievo della Sofferenza
6. Department of Clinical and Experimental Medicine, Federico II University, Naples, Italy.
7. Department, IRCCS Casa Sollievo della Sofferenza, San Giovanni Rotondo (FG), Italy.
8. CSS-Mendel Institute, Rome, Italy.

Introduction:
The article aims to investigate the association between sex and age with Apolipoprotein E (apoE) and Angiotensin-Converting Enzyme (ACE) genotypes in healthy aging and longevity among a population-based sample of individuals living in Southern Italy.

Materials and Methods:
A total of 1710 individuals were included in the study, 1344 were healthy Caucasians, and 64 were centenarians. The ACE genotypes were identified using previously described methods. The observed genotype frequencies were compared with Hardy-Weinberg frequencies using Arlequin Software. Relative allele frequencies were estimated by the gene counting method, and the 95% confidence intervals (CIs) expressing variances of the estimated allele frequencies were also calculated.

Results:
Compared to participants younger than 60 years, a significantly higher frequency of the apoE/e2 was observed in men aged 60-90 years (p<.001) and in centenarians (p<.001). Logistic regression analysis confirmed this outcome in both participants aged 60-90 years (odds ratio [OR] 1.897; 95% confidence interval [CI], 1.227-2.931) and centenarians (OR 3.263; 95% CI, 1.860-5.722). A further significant association of ACE/D allele and age was observed in centenarians (OR 2.135; 95% CI, 1.253-3.636). No relationship between apoE and ACE polymorphism was found.

Conclusion:
In conclusion, the findings suggest a role of sex in the association of apoE and ACE gene polymorphisms with healthy aging and longevity.Other chronic neurological diseases or psychiatric syndromes with cognitive impairment (16), and (c) no cerebrovascular disease, nephropathy, or end-stage renal disease, and no severe functional limitations. We considered genotypes according to the presence of at least one allele. For the ApoE polymorphism, we reported e2 (e2/) and e4 (e4/) carriers. e2 carriers were defined as those participants showing at least one e2 allele, that is, having an e2/e2, an e2/e3, or an e2/e4 genotype. Similarly, e4 carriers were defined as those participants showing at least one e4 allele, that is, having an e4/e2, an e4/e3, or an e4/e4 genotype. For ACE polymorphism, we reported D (D/) and I (I/) carriers. D carriers were defined as those participants showing at least one D allele, that is, having a D/D or a D/I genotype. Similarly, I carriers were those participants showing at least one I allele, that is, having an I/I or an I/D genotype.

ApoE and ACE Genotyping

Genomic DNA was manually purified from 4 mL of frozen blood samples by organic protein extraction and ethanol precipitation according to standard methods (17). The ApoE and ACE genotypes were determined by polymerase chain reaction (PCR) and agarose gel electrophoresis as follows.

Pearson's chi-square test and Fisher's Exact test were used to compare genotype and estimated allele frequencies. Binary logistic regression analysis was used to verify the associations between ApoE and ACE alleles with age and sex, as well as to verify interactions between ApoE and ACE alleles. All statistical analyses were made with SPSS statistical software (version 10.1.3; SPSS Inc., Chicago, IL). A value of p < .05 was considered to be statistically significant.

Briefly, the two transitions C3937 ! T (Arg112 ! Cys) and C4075 ! T (Arg158 ! Cys) resulting in the ApoE polymorphism, that is, the two single nucleotide polymorphisms rs429358 and rs7412, abolished two HhaI restriction sites (GCGC!GTGC), corresponding to codons 112 and 158 of the protein. We used the forward primer described by Houlston and colleagues (18) (TCC AAG GAG CTG CAG GCG GCG CA) and a reverse primer modified from Emi and colleagues (19) (CCC TCG CGG GCC CCG GCC TGG TAC AC). Primers were synthesized by Invitrogen (Invitrogen Corporation, Carlsbad, CA). A total of 100 ng of genomic DNA was amplified on an Applied Biosystems GeneAmp PCR System 9700 (Applied Biosystems Inc., Foster City, CA). Amplification conditions were 94°C for 2 minutes, followed by 35 cycles at 96°C for 30 seconds, 65°C for 45 seconds, and 72°C for 45 seconds. Reaction buffer included 1.5 U of Taq DNA polymerase (Platinum Taq; Invitrogen Corporation), 10 pmol of each primer, 100 μM of each deoxynucleotide triphosphate, and 1 mM MgCl2.

RESULTS