Study of cardiovascular disease biomarkers among tobacco consumers, part 2: biomarkers of biological effect

Figures & data

Table 1. Hematologic biomarkers.

		LS means^a			Group comparison p values
Biomarker	Age (years)	SMK	MSC	NTC	SMK versus MSC	SMK versus NTC	MSC versus NTC
Hemoglobin (g dL^−1)	All ages	15.91	15.53	15.35	0.1137	0.0027	0.9126
Hematocrit (%)	All ages	46.10	45.25	44.50	0.2409	0.0012	0.3678
Platelet count (thou μL^−1)	All ages	251.93	256.59	244.95	1.0000	1.0000	0.7050
RBC count (mil μL^−1)	All ages	5.06	5.08	5.06	1.0000	1.0000	1.0000
RDW (%)	All ages	12.81	12.85	12.77	1.0000	1.0000	1.0000
MCH (pg)	All ages	31.51	30.59	30.38	0.0007	<0.0001	1.0000
MCHC (g dL^−1)^b	26–31	34.49	34.32	34.24	1.0000	0.5217	1.0000
	32–37	34.59	34.61	34.84	1.0000	0.5565	0.7221
	38–43	34.40	34.49	34.48	1.0000	1.0000	1.0000
	44–49	34.55	33.83	34.44	0.0072	1.0000	0.0282
	All ages	34.51	34.31	34.50	0.1635	1.0000	0.1965
MCV (fL)	All ages	91.30	89.16	88.05	0.0036	<0.0001	0.2728
MPV (fL)	All ages	8.43	8.14	8.23	0.2931	0.6435	1.0000
WBC count (thou μL^−1)	All ages	8.48	7.53	6.75	0.0139	<0.0001	0.0591
Basophils (%)	All ages	0.44	0.45	0.52	1.0000	0.1383	0.3567
Eosinophils (%)	All ages	2.07	2.16	2.86	1.0000	0.0606	0.1767
Lymphocytes (%)	All ages	27.80	28.19	30.19	1.0000	0.1797	0.4422
Monocytes (%)	All ages	6.81	6.74	7.53	1.0000	0.1410	0.1383
Neutrophils (%)	All ages	62.89	62.45	58.90	1.0000	0.0252	0.0906

^aLeast square means.

^bAge main effect (p < 0.05).

Table 2. Serum lipid biomarkers.

		LS means^a			Group comparison p values
Biomarker	Age (years)	SMK	MSC	NTC	SMK versus MSC	SMK versus NTC	MSC versus NTC
Total cholesterol (mg dL^−1)^b	26–31	185.53	182.71	178.87	1.0000	1.0000	1.000
	32–37	199.93	194.33	214.00	1.0000	0.7515	0.3915
	38–43	202.27	199.40	206.07	1.0000	1.0000	1.0000
	44–49	188.20	208.29	222.60	0.5727	0.0162	1.0000
	All ages	193.98	196.18	205.38	1.0000	0.1902	0.5022
LDL-C (mg dL^−1)^b	26–31	117.93	112.64	108.80	1.0000	1.0000	1.0000
	32–37	126.67	110.82	134.20	0.5475	1.0000	0.1500
	38–43	126.13	125.40	130.87	1.0000	1.0000	1.0000
	44–49	120.13	136.29	150.60	0.7152	0.0174	0.8874
	All ages	122.72	121.29	131.12	1.0000	0.3744	0.3027
HDL-C (mg dL^−1)	All ages	41.93	42.30	46.83	1.0000	0.0858	0.1866
VLDL-C (mg dL^−1)	All ages	29.42	31.71	27.42	1.0000	1.0000	0.3654
ox-LDL (U L^−1)	All ages	76.42	78.93	77.40	1.0000	1.0000	1.0000
Lp(a) (mg dL^−1)	All ages	24.98	21.48	30.41	1.0000	1.0000	1.0000
Triglycerides (mg dL^−1)^b	26–31	156.47	156.57	125.40	1.0000	0.7338	0.7536
	32–37	153.33	222.17	157.87	0.0474	1.0000	0.072
	38–43	158.40	160.33	146.00	1.0000	1.0000	1.0000
	44–49	120.00	120.14	118.07	1.0000	1.0000	1.0000
	All ages	147.05	164.80	136.83	0.6645	1.0000	0.1647
Apo A1 (mg dL^−1)^b	26–31	110.47	113.07	124.87	1.0000	0.2310	0.4617
	32–37	124.33	118.92	138.87	1.0000	0.2229	0.0639
	38–43	127.93	123.93	132.87	1.0000	1.0000	0.8133
	44–49	126.33	130.43	139.60	1.0000	0.3090	1.0000
	All ages	122.27	121.59	134.05	1.0000	0.0123	0.0156
Apo A2 (mg dL^−1)	All ages	37.73	38.42	41.27	1.0000	0.0054	0.0600
Apo B100 (mg dL^−1)	All ages	92.40	93.05	94.20	1.0000	1.0000	1.0000

^aLeast square means. ^bAge main effect (p < 0.05).

Table 3. Serum BoBE.

		LS means^a			Group comparison p values
Biomarker	Age (years)	SMK	MSC	NTC	SMK versus MSC	SMK versus NTC	MSC versus NTC
AAT (mg dL^−1)	All ages	132.98	119.59	120.70	<0.0001	<0.0001	1.0000
Folate (nmol L^−1)	All ages	24.15	25.27	28.84	1.0000	0.0054	0.0840
TIMP1 (ng mL^−1)	All ages	82.88	77.88	73.39	0.1032	<0.0001	0.0942
Fibrinogen (mg dL^−1)	All ages	386.38	359.02	360.37	0.1095	0.0918	1.0000
IL-8 (pg mL^−1)	All ages	15.10	13.18	10.69	0.0924	<0.0001	0.0165
IL-12(p40) (ng mL^−1)	All ages	0.35	0.39	0.38	0.9201	1.0000	1.0000
IL-12(p70) (pg mL^−1)	All ages	60.53	50.26	49.02	<0.0001	<0.0001	1.0000
sICAM-1 (ng mL^−1)^b	26–31	149.87	128.00	109.67	0.0921	0.0003	0.2082
	32–37	162.53	124.58	109.87	0.0012	<0.0001	0.4830
	38–43	143.00	126.93	105.93	0.3147	0.0006	0.1038
	44–49	124.80	99.57	115.80	0.1281	1.0000	0.5721
	All ages	145.05	119.77	110.32	<0.0001	<0.0001	0.2385
sVCAM (ng mL^−1)	All ages	580.35	567.78	513.12	1.0000	0.0123	0.0933
sSELE (ng mL^−1)^b	All ages	47.33	46.96	44.47	1.0000	1.0000	1.0000
sSELP (ng mL^−1)	All ages	64.33	62.54	55.46	1.0000	0.0738	0.2919
MCP-1 (pg mL^−1)	All ages	397.48	356.93	286.97	0.3696	<0.0001	0.0249
sCD40L (ng mL^−1)	All ages	0.72	0.76	0.70	1.0000	1.0000	1.0000
VEGF (pg mL^−1)^b	26–31	594.27	589.14	445.37	1.0000	0.4203	0.4842
	32–37	626.80	552.75	256.87	1.0000	0.0009	0.0180
	38–43	462.87	601.60	499.00	0.5040	1.0000	0.9216
	44–49	522.67	644.29	582.80	1.0000	1.0000	1.0000
	All ages	551.65	596.94	446.10	1.0000	0.1101	0.0189
PDGF (pg mL^−1)	26–31	1799.5	3909.3	3942.2	0.0002	0.0001	0.9532
	32–37	2963.3	3204.2	1821.7	0.6803	0.0396	0.0190
	38–43	3155.3	3304.9	2689.3	0.7861	0.3982	0.2649
	44–49	2769.3	3504.3	2457.8	0.2882	0.5720	0.1312
	All ages	2671.9	3480.7	2727.8	0.0228	1.0000	0.0387
hsCRP (μg mL^−1)	All ages	3.06	2.71	1.79	1.0000	0.1413	0.5583
RANTES (ng mL^−1)	All ages	16.05	17.26	15.50	1.0000	1.0000	0.8643
MMP-3 (ng mL^−1)	All ages	5.24	5.24	4.96	1.0000	1.0000	1.0000
vWF (μg mL^−1)	All ages	27.18	22.95	19.68	0.0627	<0.0001	0.2220
Arg (μg mL^−1)	All ages	13.46	12.21	12.80	0.2289	0.9126	1.0000
ADMA (μg mL^−1)^b	26–31	0.27	0.17	0.25	<0.0001	0.2805	0.0005
	32–37	0.16	0.19	0.22	0.3652	0.0068	0.0954
	38–43	0.15	0.20	0.18	0.0498	0.2928	0.3583
	44–49	0.23	0.18	0.18	0.0895	0.0442	0.9284
	All ages	0.20	0.18	0.21	0.2199	1.0000	0.1070
SDMA (μg mL^−1)	All ages	0.43	0.42	0.49	1.0000	0.2094	0.1314
L-NMMA (μg mL^−1)^b	26–31	0.064	0.044	0.053	<0.0001	0.0003	0.0066
	32–37	0.048	0.047	0.051	0.7508	0.4169	0.2793
	38–43	0.046	0.048	0.047	0.5076	0.7162	0.7646
	44–49	0.055	0.047	0.049	0.0538	0.0809	0.5886
	All ages	0.053	0.047	0.050	0.0003	0.1011	0.1443
Hcy (μg mL^−1)	All ages	0.07	0.07	0.06	1.0000	0.6363	1.0000
Cit (μg mL^−1)^b	26–31	4.54	5.50	4.92	0.1980	1.0000	0.8070
	32–37	6.02	6.27	5.18	1.0000	0.3069	0.1407
	38–43	5.47	4.95	5.78	0.9180	1.0000	0.3150
	44–49	6.13	5.79	5.88	1.0000	1.0000	1.0000
	All ages	6.13	5.63	5.44	1.0000	1.0000	1.0000
Met (μg mL^−1)		3.79	3.78	4.27	1.0000	0.0624	0.0858

IL-1β, IL-6, TNFα, INF-γ, MMP-2 and MMP-9 were not analyzed because values were below the limit of detection.

^aLeast square means. ^bAge main effect (p < 0.05).

Figure 1. PCA. The case-wise scores of the top two principal components were plotted onto proximity maps. The top candidate analytes for BoBE that best differentiated the three study groups in this study were IL-12p70, ICAM-1 and IL-8. The concept of differentiation, or separation, in this analysis means that these biomarkers correlate and vary in such a way that identifies SMK from non-smokers (NTC and MSC) at the individual level, as opposed to an aggregate or mean level. (A) The top three biomarkers plotted for all three groups, showing a moderate separation for SMK and little separation of MSC and NTC overall. (B) A clearer separation of SMK to NTC with few individual exceptions. (C) Comparatively greater amount of overlap between SMK and MSC, while in (D), no separation is observed between MSC and NTC.

Table 4. Urine BoBE.

		LS means^a			Group comparison p values
Biomarker	Age (years)	SMK	MSC	NTC	SMK versus MSC	SMK versus NTC	MSC versus NTC
Day 1
iPF2α-III (pg mg^−1 CRE)	All ages	447.61	207.74	215.29	<0.0001	<0.0001	1.0000
iPF2α-VI (pg mg^−1 CRE)	All ages	3257.27	2403.48	2745.55	0.0423	0.1761	0.9771
TXB2 (ng mg^−1 CRE)	All ages	0.45	0.31	0.29	<0.0001	<0.0001	1.0000
Day 2
iPF2α-III (pg mg^−1 CRE)	All ages	407.01	201.80	191.68	<0.0001	<0.0001	1.0000
iPF2α-VI (pg mg^−1 CRE)	All ages	2760.76	2050.14	2267.35	0.0006	0.0159	0.7455
TXB2 (ng mg^−1 CRE)	All ages	0.66	0.48	0.39	0.0002	<0.0001	0.1137

^aLeast square means. CRE, creatinine.

Table 5. Physiological assessments of biological effect.

		LS means^a			Cohort comparison p values
Assessment	Age (Years)	SMK	MSC	NTC	SMK versus MSC	SMK versus NTC	MSC versus NTC
Day 1
FMD (%)	All ages	8.59	6.57	8.64	0.4134	1.0000	0.3895
ABI	All ages	1.12	1.14	1.15	0.3252	0.0056	0.5830
ECO (ppm)	All ages	34.32	1.63	1.67	<0.0001	<0.0001	1.0000
Day 2
FMD (%)	All ages	10.19	9.97	8.26	1.0000	0.5935	0.8773
ABI	All ages	1.16	1.15	1.17	1.0000	1.0000	0.5853
ECO (ppm)	All ages	12.87	2.12	1.75	<0.0001	<0.0001	1.0000
CIMT (mm)^b	26–31	0.56	0.58	0.56	1.0000	1.0000	1.0000
	32–37	0.61	0.63	0.61	1.0000	1.0000	1.0000
	38–43	0.65	0.66	0.63	1.0000	1.0000	0.6399
	44–49	0.73	0.63	0.69	0.0174	0.3822	0.3501
	All ages	0.64	0.63	0.62	1.0000	0.6381	1.0000

^aLeast square means. ^bAge main effect (p < 0.05).