Supplemental Methods
Biomarker Analysis
Venous blood was collected in tubes containing EDTA and was maintained at 40 C for 4
hours or less. Plasma was then removed and frozen at -800 C until assays were performed.
High sensitivity C-reactive protein (hs-CRP) was analyzed on thawed samples by the
Roche/Hitachi 912 System, Tina-quant assay (Roche Diagnostics, Indianapolis, IN). The
detection range of this assay was 0.1 mg/liter to 20 mg/liter. Values above the upper limit were
designated as 20 mg/liter.(1) Cystatin-C was analyzed by a BNII nephelometer (Dade Behring,
Inc, Deerfield, Ill; now Siemens Healthcare Diagnostics, Inc) with a particle enhanced
immunonephelometric assay (N Latex Cystatin C, Dade Behring, Inc). Monoclonal antibodies
coated on polystyrene particles that agglutinate were used to magnify the amount of scattered
light based on cystatin-C concentrations. The detection range of this assay was 0.195 mg/L to
7.330 mg/L.(2) NT-pro-BNP was analyzed by Elecsys pro-BNP platform (Roche Diagnostics,
Indianapolis, Indiana).(3) Adiponectin level was measured by a commercial sandwich enzymelinked immunosorbent assay (Millipore, Billerica, MA, USA).(4) Leptin was analyzed by a
commercially available radioimmunoassay (Linco Research Inc., St. Charles, Missouri). The
lowest detectable level was 0.5 μg/L, and all lower values were designated as 0.5 μg/L.(5) IL-6
concentrations were determined using a Luminex sandwich assay with a detection range
between 10-15,000 pg/mL. Homeostatic model assessment-insulin resistance (HOMA-IR) was
calculated from fasting glucose and insulin levels using the following formula: fasting plasma
insulin x fasting plasma glucose / 22.5.(6)
References
(1) Khera A, Vega GL, Das SR et al. Sex differences in the relationship between C-reactive
protein and body fat. J Clin Endocrinol Metab 2009;94:3251-8.
(2) Patel PC, Ayers CR, Murphy SA et al. Association of cystatin C with left ventricular structure
and function: the Dallas Heart Study. Circulation Heart failure 2009;2:98-104.
(3) Das SR, Abdullah SM, Leonard D et al. Association between renal function and circulating
levels of natriuretic peptides (from the Dallas Heart Study). The American journal of cardiology
2008;102:1394-8.
(4) Turer AT, Khera A, Ayers CR et al. Adipose tissue mass and location affect circulating
adiponectin levels. Diabetologia 2011;54:2515-24.
(5) Abdullah SM, Khera A, Leonard D et al. Sex differences in the association between leptin
and CRP: results from the Dallas Heart Study. Atherosclerosis 2007;195:404-10.
(6) Wallace TM, Levy JC, Matthews DR. Use and abuse of HOMA modeling. Diabetes Care
2004;27:1487-95.
Supplemental Methods
Statistical Analysis
Variance inflation factors and correlations coefficients of the adipose tissue and
anthropomorphic variables used in the multivariate model.
(a) Spearman correlation matrix of BMI and adiposity measures:
RP fat
BMI
IP fat
VAT
SAT
RP fat
BMI
0.57
p<.0001
IP fat
0.84
p<.0001
0.63
p<.0001
VAT
0.93
p<.0001
0.63
0.98
p<.0001 p<.0001
SAT
0.43
p<.0001
0.86
0.52
p<.0001 p<.0001
0.51
p<.0001
LBF
0.13
p=0.0001
0.67
0.21
p<.0001 p<.0001
0.19
0.81
p<.0001 p<.0001
(b) Variance inflation factors from the full model
Adiposity
Measure
VIF
BMI
6.8
RP fat
1.7
IP fat
2.1
SAT
5.9
Liver fat
1.4
LBF
4.8
LBF
Supplemental Figure 1:Forest plot demonstrating that the relative risk (95% CI) of VAT on the
development of hypertension is independent of circulating markers of renal function (cystatin-C),
inflammatory markers (hs-CRP, IL-6), adipocytokines (leptin, adiponectin), insulin-resistance
(HOMA-IR) or NT-pro-BNP.
Supplemental Figure 2: An increased incidence of hypertension was noted across sex- and
race-specific tertiles of (a) VAT or (b) RP even among those participants among the lowest
baseline sex- and race-specific quartile of SAT.