Figure 1. A, Region in the apoC-III promoter containing a functional insulin response element. Dotted line represents functional insulin-element (C3IRE) from positions -490 to -449 (11,12). Solid line indicates the position of the sequence similar to the rat phospho-carboxykinase (PEPCK) insulin/phorbol ester response element (9,10), mapped to positions -463 to -454. Asterisks show two proximal polymorphic sites located inside the C3IRE. T/C polymorphism at position -455 is located inside the rat PEPCK insulin/phorbol ester response element-like region. Sequence of the entire area can be found in the GenBank: access number X13367. B, Amplification of the mismatch chain reaction (PCR) product with the formation of the MboI recognition site. Genomic DNA (i) is used for PCR, which generates -457G rather than -457C nucleotide via site-directed mutagenesis (ii). PCR product was digested with MboI restriction endonuclease. Thymine at the -455 position (T allele) forms the authentic recognition site for MboI (fl GATC) (iii); in the case of cytosine at position -455 (C allele), the restriction site is absent (iv). C, An example of ethidium bromide-stained polyacrylamide gel from which allele determinations were made. 3, 4, and 5 are samples after digestion with MboI (lane 3, heterozygous -455T/C; lane 4, homozygous -455C; lane 5, homozygous -455T); 2, PCR product; lanes 1 and 6, the marker pBR322/AluI. bp 5 base pair.

Interestingly, the observed distribution of alleles in young (6-17 years) St. Petersburg citizens closely matched those in the group of Italian schoolchildren (11-13 years; 0.596/0.404) (7) and young (9-17 years) Canadian Indians (0.556/0.444) (19), suggesting that the initial allele distribution is similar in different populations (Table 1).

0.619, respectively, suggesting that a gradual loss of the T-allele proportion, with a relative accumulation of the C-allele proportion occurs with age (p<0.05 and p<0.02, respectively, and p<0.002 for pooled groups 2 & 3 vs group 4). The reliability of subject group size was confirmed with aThere are several corrections to be made in the given text:

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There was a statistically significant difference in the distribution of alleles for group 3 (90-106 years old; mean age, 94.5 years) versus group 4 (control; p, .02). Table 3 shows the allele distribution as a function of age and arterial hypertension. The close distribution of allele frequencies in the group and those of subjects aged 70 to 79 years old (youngest group of elderly subjects studied) make the distribution of cross-sectionally studied groups reliable.

Geriatric populations allow assessment of polymorphic sites for longevity and survival. Control groups consisting of children provide valuable information regarding the original distribution of alleles of interest in a particular population. The use of young and older age groups in our study gave us the ability to detect the effect of relative accumulation with age of the apoC-III gene T-455C polymorphism C allele.

Although we show that in this population none of the polymorphism alleles is associated with IHD or AH, the question of the possible mechanisms involved is still open and should stimulate further investigations.

St. Petersburg has one of the most advanced systems of geriatric help in Russia, which, together with the size of its population, made our study possible. However, longitudinal cohort studies covering these wide age ranges are quite hard to perform.

We have to admit that the initial frequencies of respective alleles and genotypes could differ slightly in the elderly and children's groups, as there is no way to ensure genetic solidarity of these groups when taking into account the losses the St. Petersburg population suffered during the Siege of 1941 to 1944 and the resulting changes in population structure.

The proposed method of -455 apoC-III polymorphism identification based on the mismatch PCR-mediated site-directed mutagenesis and restriction fragment length polymorphism seems to be time- and cost-reducing compared with other nonradioactive methods, such as allele-specific amplification and allele-specific oligonucleotide hybridization, described previously.

Table 4 shows the allele distribution as a function of age and ischemic heart disease.

The role of insulin resistance and the sympathoadrenal system in metabolic abnormalities. N Engl J Med.; : 

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