Supplemental Material

Association of Parental Obesity with Concentrations of Select Systemic
Biomarkers in Non-obese Offspring: The Framingham Heart Study
Wolfgang Lieb, MD; Michael J. Pencina, PhD; Katherine J. Lanier, MA; Geoffrey H. Tofler,
MD; Daniel Levy, MD; Caroline S. Fox, MD, MPH; Thomas J. Wang, MD, MPH; Ralph B.
D’Agostino Sr., PhD; Ramachandran S. Vasan, MD
From the Framingham Heart Study, 73 Mount Wayte Avenue, Framingham, MA (WL, MJP,
DL, CSF, TJW, RSV); Department of Mathematics (MJP, KJL, RBDA Sr.), and Preventive
Medicine and Cardiology Sections (RSV), Boston University School of Medicine, Boston,
MA; Royal North Shore Hospital, Sydney, Australia (GHT); The Center for Population
Studies of the National Heart, Lung, and Blood Institute, Bethesda, MD (DL, CSF);
Endocrinology Division, Brigham and Women's Hospital (CSF) and Division of Cardiology,
Department of Medicine, Massachusetts General Hospital, Harvard Medical School, Boston,
Study sample. In 1948, 2336 men and 2873 women were enrolled in the original cohort of
the Framingham Heart Study, a longitudinal epidemiological study to investigate risk factors
for cardiovascular disease. Starting in 1971, at total of 5,124 offspring of the original cohort
and their spouses were enrolled in the Framingham Offspring study (1). Offspring study
participants who were non-obese (BMI <30 kg/m²), free of prevalent cardiovascular disease at
examination cycle 6 (1995-1998) and who had both parents in the original cohort were
eligible for the present analysis (n=1,272). Offspring who did not have both parents in the
original cohort were older, and had higher systolic blood pressure and lower total cholesterol
levels. The study was conducted in accordance with the Helsinki Declaration and the study
protocols were approved by the Boston University Medical Center Institutional Review Board.
All participants gave written informed consent.
Biomarker Measurements. We analyzed a set of 7 biomarkers that have been previously
associated with adiposity-related traits or with obesity in our cohort. At examination cycle 6,
blood was drawn after an overnight fast with the participant being in a supine position for 5 to
10 minutes. After venipuncture, blood was centrifuged immediately, and plasma and serum
were stored at -80°C until the biomarkers were measured. High-sensitivity CRP was
determined using a nephelometer (Dade Behring BN100). Serum aldosterone was measured
with a radioimmunoassay (Quest Diagnostics) and plasma renin was measured using an
immunochemiluminometric assay (Nichols assay, Quest Diagnostics). Plasma B-type
natriuretic peptide and N-terminal proatrial natriuretic peptide were measured with highsensitivity immunoradiometric assays (Shionogi, Japan). Plasma levels of fibrinogen were
assessed with the Claus method and PAI-1 antigen was measured by ELISA (TintElize PAI-1,
Biopool AB). The average interassay coefficients of variation for the biomarkers were as
follows: C-reactive protein, 2.2%; aldosterone, 4.0% (for high concentrations) and 9.8% (for
low concentrations); renin, 2.0% (for high concentrations) and 10.0% (for low concentrations);
Plasma B-type natriuretic peptide, 12.2%; N-terminal proatrial natriuretic peptide, 12.7%;
fibrinogen, 2.6% and PAI-1, 7.7%.
1. Kannel WB, Feinleib M, McNamara PM, Garrison RJ, Castelli,WP: An investigation of
coronary heart disease in families. The Framingham offspring study. Am.J.Epidemiol.
110:281-290, 1979