Table 3 Genotypic and allelic frequencies of the APOA4- HincII-RFLP in samples of individuals (males, 3a; females, 3b) of different ± × ages. Relative frequencies standard errors ( 100) are given in parentheses. A/B denotes Absence/Presence of the restriction site. HWE denotes values expected at Hardy-Weinberg Equilibrium

3a Males (n = 327) 18–45 years 46–80 years 81–109 years Genotypes Counts (120) HWE Counts (155) HWE Counts (52) ± ± ± AA 2 (1.7 1.2) 2.6 1 (0.6 0.6) 3.1 3 (5.8 3.2) 2.1 ± ± ± AP 31 (25.8 4.0) 29.9 42 (27.1 3.6) 37.8 15 (28.8 6.3) 16.8 ± ± ± PP 87 (72.5 4.1) 87.6 112 (72.3 3.6) 114.1 34 (65.4 6.6) 33.1 Alleles Counts (240) Counts (310) Counts (104) A 35 (14.6) 44 (14.2) 21 (20.2) ± ± ± P 205 (85.4 2.3) 266 (85.8 2.0) 83 (79.8 3.9) 3b Females (n = 473) 18–45 years 46–80 years 81–109 years Genotypes Counts (184) HWE Counts (112) HWE Counts (177) ± ± ± AA 2 (1.1 0.8) 4.1 1 (0.9 0.9) 1.2 1 (0.6 0.6) 3.2 ± ± ± AP 51 (27.7 3.3) 46.8 21 (18.8 3.7) 20.6 46 (26.0 3.3) 41.5 ± ± ± PP 131 (71.2 3.3) 133.1 90 (80.4 3.8) 90.2 130 (73.4 3.3) 132.3 Alleles Counts (368) Counts (224) Counts (354) A 55 (14.9) 23 (10.3) 48 (13.6) ± ± ± P 313 (85.1 1.9) 201 (89.7 2.0) 306 (86.4 1.8)

 Table 4 Haplotypes of APOA1-MspI-RFLP (5 ) and APOC3- 3!) SstI-RFLP (3 ) in samples of individuals of different ages. A/P indicates Absence/Presence of the restriction site. Haplotypic fre- × quencies ( 100) are estimated by Maximum Likelihood method. Standard errors are in parentheses

 Table 5 Haplotypes of APOC3-SstI-RFLP (5 ) and APOA4- ae3/) HincII-RFLP (3 ) in samples of individuals of different ages. A/P indicates Absence/Presence of the restriction site. Haplotypic fre- × quencies ( 100) are estimated by Maximum Likelihood method. Standard errors are in parentheses

Males Haplotypes 18–45 years 46–80 years 81–109 years     ± ± ± 5 AA3 6.9 ( 1.7) 5.6 ( 1.3) 11.7 ( 3.2)    ± ± ± 5 AP3 83.5 ( 2.4) 83.7 ( 2.1) 76.8 ( 4.2)    ± ± ± 5 PA3 7.6 ( 1.7) 8.6 ( 1.6) 10.5 ( 3.1)    ± ± ± 5 PP3 2.0 ( 0.9) 2.1 ( 0.8) 1.1 ( 1.1) Females Haplotypes 18–45 years 46–80 years 81–109 years  3! ± ± ± 5 AA3 5.3 ( 1.2) 5.5 ( 1.6) 6.5 ( 1.3)    ± ± ± 5 AP3 83.7 ( 2.0) 86.9 ( 2.3) 86.1 ( 1.8)    ± ± ± 5 PA3 9.8 ( 1.6) 4.8 ( 1.5) 7.0 ( 1.4)    ± ± ± 5 PP3 1.2 ( 0.6) 2.8 ( 1.2) 0.3 ( 0.3)

Males Haplotypes 18–45 years 46–80 years 81–109 years  3! ± ± ± 5 AA3 26.4 ( 3.1) 30.7( 2.8) 18.3 ( 3.8)   ± ± 5 AP3 1.3 ( 1.1) 1.9 ( 1.1) 0   ± ± ± 5 PA3 64.0 ( 3.3) 58.7 ( 2.9) 70.2 ( 4.6)   ± ± ± 5 PP3 8.3 ( 2.0) 8.7 ( 1.7) 11.5 ( 3.2) Females Haplotypes 18–45 years 46–80 years 81–109 years  3! ± ± ± 5 AA3 20.8 ( 2.1) 26.3 ( 3.0) 25.4 ( 2.3)   ± 5 AP3 0 0.9 ( 0.9) 0   ± ± ± 5 PA3 68.2 ( 2.5) 66.1 ( 3.2) 67.2 ( 2.6)   ± ± ± 5 PP3 11.0 ( 1.8) 6.7 ( 1.9) 7.3 ( 1.4)

significant effect was found on any of the lipidemic pa- rameters (Table 6b).

termediate age-class (allele A) had a specific LDL-C de- creasing effect. On the contrary, the allele which reached the lowest frequency in the intermediate age-class (allele P) had a specific LDL-C increasing effect. Since

On the whole, this data showed that, in males the allele which reached the highest frequency in the in

 C University College London 2003

S. Garasto et al.

α α Table 6 Allele average effects and estimated for APOA1 A and P alleles respectively in 45–80 year old subjects. Average effects A P are expressed in mg/dL. Confidence intervals (c.i.) are computed by bootstrap

6a Males α α 95% c.i. 95% c.i. A P − − − Total Cholesterol 6.25 14.83 : +2.31 +3.74 1.31 : +8.73 − − − HDL-Cholesterol +0.74 1.77 : +0.75 0.42 1.82 : +1.03 − − − LDL-Cholesterol 8.19 15.99 : 0.26 +4.70 +0.14 : +9.23 − − − Triglycerides (Log ) +0.02 0.02 : + 0.07 0.01 0.04 : + 0.01 10 6b Females α α 95% c.i. 95% c.i. A P − − Total Cholesterol +0.09 10.70 : +10.21 +0.05 6.70 : +5.50 − − − HDL-Cholesterol +2.0 1.68 : +5.58 1.08 3.13 : +0.84 − − − LDL-Cholesterol 0.51 11.80 : +10.60 +0.28 5.60 : +6.10 − − Triglycerides (Log ) 0.07 0.05 : +0.06 +0.01 0.03 : +0.03 10

the samples we were analysing had been selected for good health and normal lipidemic parameters, this finding was reasonable. In line with these results we found that the frequency of the P allele was significantly higher in 46–80 year old male patients affected by CD with high level of serum LDL-C (CDL patients, aver- age serum LDL-C = 206.53 mg/dL, with 29.66 mg/dL SD) than in sex- and age-matched healthy persons. In ± fact, the P allele frequency was 0.814 ( 0.047 standard ± error) in CDL patients versus 0.674 ( 0.027 standard error) in age-matched healthy males. As expected, a dif- ferent result was found in 46–80 year old male patients affected by CD with low levels of serum HDL-C (CDH patients, average serum HDL-C = 27.79 mg/dL, with 3.15 mg/dL SD), where the P allele frequency was 0.679 ± ( 0.051 standard error).