concentrations, which indicate a worse metabolic profile. Furthermore, no beneficial effects of the ER22/23EK polymorphism on the prevalence of cardiovascular pathologies, allcause and cardiovas- cular mortality were found. These result are at odds with results from the previously published studies (van Rossum et al. 2002, 2004a). It could be, that the discrepancy between these studies has arisen due to the age difference of the subjects. In our study, par- ticipants aged 85–90 years were analyzed compared to subjects aged 67–77 years in the other studies (van Rossum et al. 2002, 2004a). The analyzed polymor- phisms might have an effect before a certain age, and therefore the Leiden 85-plus cohort could consists of survivors. To test this, allele and genotype frequen- cies between the elderly and young participants in this study, and also between a young study group from the published literature (van Rossum et al. 2004b) were compared, revealing no differences. Thus, in contrast to an earlier report of an age- dependent enrichment for the ER22/23EK variation (van Rossum et al. 2002), no such enrichment was observed in our study.

mate several phenotypes in one cohort, and the prospective analysis with a high number of deaths, including the high number of cardiovascular deaths, during the follow-up. A weakness of the study is the lack of data on HPA-axis reactivity or on other parameters reflecting the stress system activity. However, in this aspect, we rely on the information presented in the published literature.

In conclusion, the results of this study show that in old age the ER22/23EK and N363S polymorphisms but not the BclI SNP in the GR gene have an influence on metabolic profile. However, the noticed differ- ences in metabolic profile do not affect the prevalence of cardiovascular pathologies and the risks for all- cause and cardiovascular disease mortality at old age.

come from the participant’s inclusion criteria. In order to apply similar inclusion criteria used in the other studies, only independently living participants were analyzed. However, this did not change our results, as well as stratification for gender. A more likely explanation could be that some of the obser- vations have arisen by chance, leading to false neg- ative or false positive associations. Recently it has been suggested that the role of genetic variations in common traits should be built up based on the evi- dence of many studies. Significant between-study diversity is frequent, and the results of the first study often correlate only modestly with subsequent re- search on the same association (Ioannidis et al. 2001).

Acknowledgements This work was supported by an IOP grant (Innovative Orientated Research) from the Dutch Min- istry of Economic Affairs (grant number IGE010114), and by the Centre for Medical Systems Biology (CMSB), which is a center of excellence approved by the Netherlands Genomics Initiative/Netherlands Organisation for Scientific Research (NWO). The study was also supported by a Marie Curie Fel- lowship of the European Community programme EURO- GENDIS ‘‘The Genetic Basis of Disease’’ for Maris Kuningas, under contract number QLGA-GH-00-60005-59. We thank Anton J. M. de Craen for the help in data analyses.

The results of this study raise doubts whether the analyzed polymorphisms, shown to result in either resistance or increased sensitivity to cortisol (van Rossum et al. 2002, 2003; Huizenga et al. 1998a), are really functional. For the ER22/23EK variant, it has been reported that it does not alter the in vitro capacity to activate transcription (de Lange et al. 1997) and no altered response to cortisol was ob- served (Koper et al. 1997). However, a more recent study shows, that the transcriptional activity of the ER22/23EK carriers is decreased, due to higher

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