pathogens.4 The specificity of the SELDI-TOF mass spec- trometry profile for IAI is suggested by the presence of the 10.8-kDa proteolytic fragment of IGFBP-1, which has not been described in other inflammatory conditions. During pregnancy, the decidua, fetal membranes, and amniotic fluid are rich sources of IGFBP-1.5 Thus, the presence of the unique IGFBP-1 proteolytic fragment together with the presence of calgranulins is consistent with intrauterine infection.

all cells. However, in atherosclerotic tissue its expression is markedly up-regulated, and the inflammatory media- tors produced by its activation exert various atherogenic effects.2,3 In this study, we evaluated whether prevalence of the TLR4 genotype may have an association with acute myocardial infarction (AMI) and with longevity.

Methods. We studied 105 young Sicilian men (mean age, 41 years; range, 20-46 years) consecutively admitted for AMI to the cardiac unit of Palermo University Hospital. Current smoking was present in 74% (former and current in 89%), family history of CVD in 58%, type 2 diabetes in 17%, obe- sity in 34%, hypertension in 29%, hypercholesterolemia in 61%, and hypertriglyceridemia in 43%.

We used pooled maternal serum samples to ascertain the possibility of detecting calgranulin B and IGFBP-1 proteolytic fragment noninvasively—without an amnio- centesis. In biomarker discovery, pooled samples are frequently used to optimize detection of potential bio- markers. We agree that larger prospective trials that test serum samples from patients are necessary to confirm our findings.

One control group consisted of 127 Sicilian men matched by age strata (mean age, 38 years; range, 20-50 years). These were students and staff personnel who were judged to be in good health based on their clinical history and on blood tests (complete blood cell count, erythrocyte sedimenta- tion rate, and levels of glucose, blood urea nitrogen, creati- nine, electrolytes, C-reactive protein, liver transaminases, iron, proteins, cholesterol, and triglycerides). Of these young controls, 20% were moderate smokers; no other major car- diac risk factor was present.

Michael G. Gravett, MD gravettm@ohsu.edu Department of Obstetrics & Srinivasa R. Nagalla, MD Department of Pediatrics Oregon Health & Science University Prote

enix Inc

A second control group consisted of 55 Sicilian oldest old men (mean age, 100 years; range, 96-104 years). By using the demographic lists of the registry offices of Palermo and surrounding municipalities, 166 potentially eligible oldest old men were identified in the Palermo area. Ninety-one (55%) agreed to participate. The control group was com- posed of the 55 (60%) who did not have any cardiac risk factors or major age-related diseases (eg, coronary heart dis- ease, severe cognitive impairment, severe physical impair- ment, clinically evident cancer, or renal insufficiency), al- though some had decreased auditory and visual acuity.4 Because immigration and intermarriage in Sicily have his- torically been rare, the Sicilian ethnicity of all participants was confirmed by having all 4 grandparents born in Sicily. The study was approved by the hospital ethics committee and written informed consent was obtained from all par- ticipants.

Financial Disclosure: Drs Gravett and Nagalla are the founders of ProteoGenix and retain stock options in the company.

1. Heine RP, Weisenfeld H, Mortimer L, Greig PC. Amniotic fluid defensins: po- tential markers of subclinical intrauterine infection. Clin Infect Dis. 1998;27:513-

2. SearleBC,DasariS,TurnerM,etal.High-throughputidentificationofproteins and unanticipated sequence modifications using a mass-based alignment algo- rithm for MS/MS de novo sequencing results. Anal Chem. 2004;76:2220-2230. 3. Vogl T, Propper C, Hartmann M, et al. S100A12 is expressed exclusively by granulocytesandactsindependentlyfromMRP8andMRP14.JBiolChem.1999; 274:25291-25296.

4. Roth J, Vogl T, Sorg C, Sunderkotter C. Phagocyte-specific S100 proteins: a novel group of proinflammatory molecules. Trends Immunol. 2003;24:155-158. 5. Hills FA, Gunn LK, Hardiman P, Thamaratnam S, Chard T. IGFBP-1 in the pla- centa, membranes and fetal circulation: levels at term and preterm delivery. Early

DNA was genotyped for the +896 A G (Asp299Gly) TLR4 single-nucleotide polymorphism (SNP) according to published methods.5 The Asp299Gly SNP has been shown to attenuate receptor signaling.3,5 Differences in SNP fre-   quency among the groups were evaluated by 2 test. We per- formed a logistic regression analysis to test the association of SNP with AMI after adjustment for smoking habits, fam- ily history of CVD, and the presence of type 2 diabetes, obe- sity, hypertension, hypercholesterolemia, and hypertriglyc- eridemia. A similar analysis was performed comparing young controls with oldest old controls.

Role of Toll-like Receptor 4 in Acute Myocardial Infarction and Longevity

To the Editor: A major trait characterizing centenarians is a relatively low frequency of cardiovascular disease (CVD). Since a proinflammatory genotype appears to contribute to the risk of ischemic heart disease, the prevalence of proinflammatory alleles would also be expected to be relatively low in people experiencing lon- gevity.1 Toll-like receptor 4 (TLR4), the activity of which may be modulated by genetic polymorphisms, is a signal- ing receptor in innate immune defense. It is activated by lipopolysaccharide from gram-negative bacteria and endogenous ligands, including those produced in response to tissue injury. TLR4 is expressed on virtually

Results. We found significant differences in the fre- quency of TLR genotypes among the 3 studied groups “AsP<.( (P .001). The Asp299Gly SNP was underrepresented in pa- tients with AMI and overrepresented in the oldest old men

(Reprinted) JAMA, November 17, 2004—Vol 292, No. 19 2339

Marco Caruso, MD Enrico Hoffmann, MD Department of Internal Medicine Cardiovascular and Nephro