Download Supplemental Methods and Tables

SUPPLEMENTAL METHODS
Echocardiographic Assessment
In Olmsted County, all echocardiograms are performed and interpreted in the Mayo
Clinic Echocardiographic Laboratory. M-mode, 2-dimensional, Doppler, and Doppler tissue
imaging (DTI) were performed according to guidelines of the American Society of
Echocardiography (1). Digital echocardiographic data containing a minimum of 3 consecutive
beats (5 in atrial fibrillation) were acquired and transferred to a server for storage and archiving
(ProSolv Echo Management System, Problem Solving Concepts, Carmel, Ind). In each subject,
ejection fraction was measured by M-mode using the modified Quinones formula, quantitative
2D (biplane Simpson), and semiquantitative 2D (visual estimate) as described previously (1-4).
Two-dimensional (2-D) and color Doppler imaging were performed to screen for valvular
disease. Valvular disease was defined as greater than mild stenosis or regurgitation as defined by
the American Society of Echocardiography (5, 6).
Pulsed-wave Doppler examination of mitral (before and with Valsalva maneuver) and
pulmonary venous inflow, as well as Doppler tissue imaging of the mitral annulus, was
performed in each subject. Diastolic function was categorized as normal, mildly impaired
(defined as impaired relaxation without evidence of increase filling pressures), moderately
impaired (defined a impaired relaxation associated with moderate elevation of filling pressures or
pseudonormal filling), and severely impaired (defined as advanced reduction in compliance) (79). Diastolic dysfunction was defined as moderate or severe dysfunction.
Left ventricular mass was calculated according to the Devereux formula (10) and indexed
to body surface area. The presence of left ventricular hypertrophy (LVH) was defined on the
basis of left ventricular mass index >130 g/m2 for men and >100g/m2 for women (11). The
presence of left atrial (LA) enlargement was defined as LA volume index >33 mL/m2 in men
and >30 mL/m2 in women (12).
Biomarker Assessment
Laboratory methods and performance data for creatinine, high-density lipoprotein, and
total cholesterol are outlined in Supplement Table 1.
Supplemental Table 1 Methods for Total Cholesterol, High Density Cholesterol, and Creatinine
Method
Precision (%CV)
Precision (%CV)
Reportable Range
Limit of Detection
TC
Enzymatic;
colorimetric
cholesterol
oxidase method
1.4% (mean 158 mg/dL);
0.9% (mean 106 mg/dL);
0.8% (mean 260 mg/dL)
1.0% (mean 105 mg/dL);
1.6% (mean 158 mg/dL);
1.1% (mean 255 mg/dL)
10-7500 mg/dL
Did not test actual LOD;
manufacturer stated low end is 10
mg/dL; verified with serum pool
(7.5% CV, mean 11 mg/dL)
HDL-C
Homogenous,
enzymatic
colorimetric assay
(PEG-cholesterol)
1.3% (mean 45 mg/dL);
0.7% (mean 29 mg/dL)
2.4% (mean 44 mg/dL);
2.9% (mean 28 mg/dL)
5-240 mg/dL
7.0% CV at mean 3 mg/dL
Creatinine
Enzymatic
(IDMS-traceable)
1.2% (mean 1.1 mg/dL);
0.83% (mean 5.9 mg/dL)
1.3% (mean 1.1 mg/dL);
2.3% (mean 5.8 mg/dL)
0.1-92.0 mg/dL
0.1 mg/dL (7.0% CV mean 0.15
mg/dL)
TC, total cholesterol; HDL-C, high density cholesterol; CV, coefficient of variance; LOD, limit of detection
Supplemental Table 2 Number and Percentage of Subjects in the Total Population and Specific
Disease Cohorts Above the Gender Specific 95th and 99th Percent of the Upper Reference Limit*
Above 95th % URL
Number (%)
Above 99th % URL
Number (%)
Total Population
All
Female
Male
222 (12)
144 (15)
245 (28)
36 (2)
23 (2)
21 (2)
Hypertension
All (n=514)
Female (n=278)
Male (n=236)
92 (18)
61 (22)
103 (44)
15 (2.9)
8 (2.9)
9 (3.8)
Left Ventricular Hypertrophy
All (n=184)
49 (27)
Female (n=132)
39 (30)
Male (n=52)
38 (73)
5 (3)
5 (4)
4 (8)
Systolic Dysfunction (EF < 50 %)
All (n=80)
40 (50)
Female (n=18)
12 (67)
Male (n=62)
43 (69)
8 (10)
2 (11)
6 (10)
Glomerular Filtration Rate < 60 ml/min
All (n=262)
67 (26)
Female (n=113)
44 (39)
Male (n=149)
71 (48)
7 (3)
5 (4)
4 (3)
Mild Diastolic Dysfunction
All (n=367)
Female (n=176)
Male (n=191)
7 (2)
6 (3)
5 (3)
60 (16)
49 (28)
76 (40)
Moderate/Severe Diastolic Dysfunction
All (n=126)
32 (25)
Female (n=70)
15 (21)
Male (n=56)
30 (54)
6 (5)
2 (3)
4 (7)
Coronary Artery Disease
All (n=221)
Female (n=61)
Male (n=160)
11 (5)
3 (5)
8 (5)
67 (30)
25 (4)
88 (55)
*Gender specific upper reference limit (URL) defined from the healthy reference cohort (Figure
2). The 95th and 99th percentile URL values for females were 6 and 33 ng/L and for males were 17
and 55 ng/L, respectively.
Supplemental Table 3 Upper Reference Limits in the Healthy Reference Cohort
When Restricted to a NT-proBNP less than 100 pg/ml
Females (n=230)
Males (n=257)
Combined (n=487)
95th percentile
7
17
12
99th percentile
36
55
51
Healthy reference cohort was characterized as subjects without cardiovascular risk factors and no echocardiographic
abnormalities (see methods). In the above upper reference limits, the healthy reference cohort was further restricted
to only individuals with a NT-proBNP less than 100 pg/ml.
Supplemental Table 4 Effect of Body Mass Index, Left Ventricular Mass, and Glomerular
Filtration Rate on High Sensitivity Troponin I in the Healthy Reference Cohort (n=565)
Model
Parameters Included in the Model
Regression
Coefficient (SE)
p-value
Age‡
Female Gender
Body mass index†
0.021 (0.006)
-0.662 (0.089)
0.068 (0.044)
<.001
<.001
0.128
Age‡
Female Gender
LV Mass Index†
0.018 (0.006)
-0.582 (0.102)
0.193 (0.050)
0.005
<.001
<.001
Age‡
Female Gender
Estimated GFR†
0.019 (0.006)
-0.690 (0.087)
-0.049 (0.045)
0.003
<.001
0.272
Age‡
Female Gender
Body mass index†
LV Mass Index †
Estimated GFR†
0.017 (0.007)
-0.563 (0.103)
0.044 (0.051)
0.198 (0.051)
-0.066 (0.048)
0.012
<.001
0.390
<.001
0.173
Model 1
Model 2
Model 3
Model 4
Dependent variable in all models was log hs-cTnI value+1. † Per standard deviation (SD) change [body mass index
(BMI) SD = 4.7 kg/m2, glomerular filtration rate (GFR) SD = 14.4 ml/min, and left ventricular (LV) mass index SD
14.9 g/m2]. ‡ Age, expressed per 10-year change. All models include age and gender. Model 1 includes BMI; Model
2 includes LV mass index; Model 3 includes GFR; Model 4 includes BMI, LV mass index, and GFR. See methods
for criteria of the healthy reference cohort.
Supplement Figure Legend
Supplemental Figure 1 Distribution of high sensitivity troponin I (hs-cTnI) in diseased states including left ventricular hypertrophy,
ejection fraction < 50 percent, moderate / severe diastolic dysfunction, glomerular filtration rate (GFR) < 60 ml/min, hypertension,
and coronary artery disease. Solid arrow denotes the 80th percentile upper reference limit (URL), open arrow the 95th percentile
URL, and solid triangle the 99th percentile URL. Gender combined URL were defined from the healthy reference cohort.
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