Supplementary Materials: A Preliminary Link between Hydroxylated Metabolites of Polychlorinated Biphenyls and Free Thyroxin in Humans

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S1 of S11 Supplementary Materials: A Preliminary Link between Hydroxylated Metabolites of Polychlorinated Biphenyls and Free Thyroxin in Humans Eveline Dirinck, Alin C. Dirtu, Govindan Malarvannan, Adrian Covaci, Philippe G. Jorens and Luc F. Van Gaal Table S1. PCB levels in serum (n = 180). Wet Weight Basis (ng/l) Lipid Weight Basis (ng/g Lipid) PCB28 1 0 174 0.2 0 111 PCB52 0 0 20 0 0 20 PCB74 18 2 353 2.9 0.3 122 PCB95 1 1 14 0.2 0.1 5 PCB99 22 2 121 0.2 0.1 5 PCB101 1 0 17 0.2 0.1 6 PCB105 7 2 102 1.1 0.3 18 PCB118 41 4 448 6.9 0.7 165 PCB149 5 1 39 0.7 0.2 39 PCB146 26 2 163 4.2 0.3 65 PCB153 259 16 1262 43.1 2.5 624 PCB138 135 2 703 22.5 0.3 317 PCB187 37 2 327 6.2 0.4 95 PCB183 17 2 327 2.8 0.3 69 PCB128 0 0 42 0.1 0 42 PCB167 22 2 161 3.7 0.3 66 PCB174 2 0 9 0.4 0.0 7 PCB177 11 2 64 1.8 0.2 26 PCB171 6 2 33 1.0 0.2 17 PCB172 6 2 51 1.0 0.2 18 PCB156 23 2 217 3.8 0.3 71 PCB180 175 10 1136 27 1.6 432 PCB170 69 4 436 10.5 0.7 167 PCB199 9 2 85 1.4 0.2 26 PCB196/203 15 2 82 2.3 0.2 40 PCB194 14 2 111 2.3 0.2 43 PBC206 2 1 14 0.4 0.1 7.0 PCB209 1 1 10 0.2 0.1 6.0 Results are expressed as median (minimum maximum).

S2 of S11 Table S2. PCB metabolite levels in serum (n = 180). Median Range (min max) 4 HO PCB79 0.0 (0.0 52) 4 HO PCB120 1.0 (0.3 5) 3 HO PCB118 5.0 (0.9 49) 4 HO PCB107 9.0 (1.0 77) 3 HO PCB153 3.0 (0.7 23) 4 HO PCB146 15.0 (1.0 131) 4 HO PCB127 1.0 (0.3 11) 3 HO PCB138 6.0 (0.8 51) 4 HO PCB130 0.0 (0.0 2) 4 HO PCB163 2.0 (0.7 12) 4 HO PCB187 16.0 (1.0 81) 4 HO PCB162 3.0 (0.9 17) 4 HO PCB177 0.0 (0.0 3) 3 HO PCB180 2.0 (0.6 19) 4 HO PCB172 4.0 (0.8 29) 4 HO PCB193 0.0 (0.0 4) 4 diho PCB202 0.0 (0.0 4) 4 HO PCB208 0.0 (0.0 5) Results are expressed in ng/l and represented as median (minimum maximum). Table S3. Linear regression for FT4 with lipid adjusted PCBs. PCB28 0.096 0.052 PCB52 0.094 0.053 PCB74 0.068 0.057 Gender 0.027 ( 0.118 0.063) 0.0552 Age 0.001 ( 0.005 0.002) 0.491 Smoking status 0.053 ( 0.061 0.168) 0.359 PCB28 0.038 ( 0.093 0.168) 0.568 Gender 0.030 ( 0.121 0.061) 0.518 Age 0.001 ( 0.004 0.002) 0.587 Smoking status 0.047 ( 0.066 0.161) 0.412 PCB52 0.083 ( 0.340 0.174) 0.525 Gender 0.038 ( 0.114 0.068) 0.617 Age 0.116 ( 0.007 0.002) 0.242 BMI 0.193 (0.001 0.011) 0.011 Smoking status 0.069 ( 0.060 0.166) 0.356 PCB74 0.107 ( 0.060 0.218) 0.264

S3 of S11 Table S3. Cont. PCB95 0.024 0.072 Gender 0.024 ( 0.113 0.066) 0.604 Age 0.001 ( 0.004 0.003) 0.754 BMI 0.007 (0.002 0.012) 0.004 Smoking status 0.041 ( 0.071 0.154) 0.470 PCB95 0.299 ( 0.595 0.002) 0.048 PCB99 0.024 0.071 Gender 0.026 ( 0.116 0.064) 0.566 Age 0.003 ( 0.007 0.001) 0.108 BMI 0.006 (0.001 0.011) 0.012 Smoking status 0.046 ( 0.066 0.158) 0.421 PCB99 0.143 (0.000 0.285) 0.049 PCB101 0.017 0.076 Gender 0.014 ( 0.104 0.077) 0.768 Age 0.001 ( 0.004 0.003) 0.676 BMI 0.006 (0.002 0.011) 0.009 Smoking status 0.037 ( 0.075 0.150) 0.516 PCB101 0.317 ( 0.601 0.033) 0.029 PCB105 0.109 0.050 Gender 0.028 ( 0.119 0.063) 0.547 Age 0.001 ( 0.005 0.003) 0.599 Smoking status 0.049 ( 0.065 0.164) 0.396 PCB105 0.002 ( 0.204 0.208) 0.986 PCB118 0.105 0.050 Gender 0.028 ( 0.119 0.063) 0.547 Age 0.001 ( 0.005 0.003) 0.599 Smoking status 0.049 ( 0.065 0.164) 0.396 PCB118 0.002 ( 0.204 0.208) 0.986 PCB149 0.017 0.076 Gender 0.030 ( 0.121 0.062) 0.586 Age 0.001 ( 0.005 0.004) 0.720 BMI 0.006 (0.002 0.011) 0.009 Smoking status 0.046 ( 0.069 0.161) 0.505 PCB149 0.248 ( 0.471 0.025) 0.030

S4 of S11 Table S3. Cont. PCB146 0.109 0.050 Gender 0.028 ( 0.119 0.063) 0.542 Age 0.001 ( 0.006 0.004) 0.653 BMI 0.006 (0.002 0.011) 0.011 Smoking status 0.050 ( 0.064 0.163) 0.391 PCB146 0.005 ( 0.172 0.181) 0.958 PCB153 0.101 0.051 Gender 0.028 ( 0.118 0.063) 0.547 Age 0.000 ( 0.005 0.005) 0.948 BMI 0.006 (0.001 0.011) 0.015 Smoking status 0.048 ( 0.066 0.162) 0.406 PCB153 0.034 ( 0.186 0.118) 0.660 PCB138 0.089 0.053 Gender 0.027 ( 0.118 0.063) 0.550 Age 0.000 ( 0.004 0.005) 0.960 BMI 0.006 (0.002 0.011) 0.009 Smoking status 0.048 ( 0.065 0.162) 0.401 PCB138 0.053 ( 0.196 0.090) 0.468 PCB187 0.108 0.050 Gender 0.028 ( 0.119 0.063) 0.540 Age 0.001 ( 0.006 0.004) 0.612 BMI 0.007 (0.002 0.012) 0.010 Smoking status 0.050 ( 0.064 0.163) 0.389 PCB187 0.011 ( 0.153 0.176) 0.892 PCB183 0.109 0.050 Gender 0.028 ( 0.119 0.063) 0.544 Age 0.001 ( 0.006 0.003) 0.638 Smoking status 0.049 ( 0.064 0.163) 0.392 PCB183 0.003 ( 0.183 0.188) 0.975 PCB128 0.105 0.051 Gender 0.028 ( 0.118 0.063) 0.548 Age 0.001 ( 0.004 0.002) 0.578 Smoking status 0.048 ( 0.067 0.162) 0.412 PCB128 0.040 ( 0.292 0.212) 0.754

S5 of S11 Table S3. Cont. PCB167 0.104 0.051 Gender 0.029 ( 0.120 0.062) 0.526 Age 0.000 ( 0.005 0.004) 0.861 BMI 0.006 (0.001 0.011) 0.016 Smoking status 0.046 ( 0.069 0.161) 0.434 PCB167 0.031 ( 0.208 0.147) 0.733 PCB174 0.067 0.057 Gender 0.026 ( 0.116 0.064) 0.571 Age 0.001 ( 0.004 0.002) 0.535 BMI 0.006 (0.001 0.011) 0.015 Smoking status 0.042 ( 0.071 0.156) 0.463 PCB174 0.224 ( 0.614 0.167) 0.260 PCB177 0.108 0.050 Gender 0.028 ( 0.119 0.062) 0.539 Age 0.001 ( 0.006 0.003) 0.571 BMI 0.007 (0.002 0.011) 0.008 Smoking status 0.050 ( 0.064 0.164) 0.388 PCB177 0.016 ( 0.182 0.213) 0.874 PCB171 0.103 0.051 Gender 0.028 ( 0.019 0.062) 0.540 Age 0.002 ( 0.006 0.003) 0.484 BMI 0.007 (0.002 0.011) 0.007 Smoking status 0.051 ( 0.063 0.165) 0.375 PCB171 0.047 ( 0.203 0.298) 0.711 PCB172 0.102 0.051 Gender 0.029 ( 0.120 0.062) 0.532 Age 0.002 ( 0.006 0.003) 0.459 BMI 0.007 (0.002 0.012) 0.008 Smoking status 0.052 ( 0.062 0.167) 0.368 PCB172 0.051 ( 0.180 0.182) 0.664 PCB156 0.109 0.050 Gender 0.028 ( 0.119 0.063) 0.541 Age 0.001 ( 0.006 0.004) 0.645 BMI 0.007 (0.001 0.012) 0.012 Smoking status 0.050 ( 0.064 0.164) 0.389 PCB156 0.009 ( 0.177 0.195) 0.926

S6 of S11 Table S3. Cont. PCB180 0.106 0.051 Gender 0.027 ( 0.118 0.063) 0.552 Age 0.000 ( 0.006 0.005) 0.868 BMI 0.006 (0.001 0.011) 0.020 Smoking status 0.048 ( 0.066 0.162) 0.406 PCB180 0.021 ( 0.174 0.132) 0.785 PCB170 0.107 0.050 Gender 0.028 ( 0.118 0.063) 0.549 Age 0.001 ( 0.006 0.005) 0.812 BMI 0.006 (0.001 0.011) 0.016 Smoking status 0.048 ( 0.065 0.162) 0.402 PCB170 0.016 ( 0.182 0.151) 0.851 PCB199 0.092 0.053 Gender 0.029 ( 0.120 0.062) 0.528 Age 0.002 ( 0.007 0.002) 0.366 BMI 0.007 (0.002 0.012) 0.006 Smoking status 0.054 ( 0.060 0.168) 0.351 PCB199 0.071 ( 0.134 0.275) 0.497 PCB196/203 0.107 0.051 Gender 0.028 ( 0.119 0.062) 0.539 Age 0.001 ( 0.006 0.003) 0.580 BMI 0.007 (0.002 0.012) 0.011 Smoking status 0.051 ( 0.064 0.165) 0.384 PCB196/203 0.019 ( 0.179 0.218) 0.847 PCB194 0.108 0.050 Gender 0.028 ( 0.119 0.063) 0.539 Age 0.001 ( 0.006 0.004) 0.628 BMI 0.007 (0.001 0.012) 0.014 Smoking status 0.050 ( 0.064 0.164) 0.389 PCB194 0.010 ( 0.187 0.208) 0.917 PCB206 0.106 0.051 Gender 0.028 ( 0.119 0.062) 0.538 Age 0.001 ( 0.005 0.003) 0.509 BMI 0.007 (0.002 0.011) 0.008 Smoking status 0.052 ( 0.063 0.167) 0.374 PCB206 0.042 ( 0.260 0.345) 0.785

S7 of S11 Table S3. Cont. PCB209 0.107 0.051 SumPCB 0.107 0.051 Gender 0.028 ( 0.119 0.062) 0.538 Age 0.001 ( 0.005 0.002) 0.524 BMI 0.007 (0.002 0.011) 0.008 Smoking status 0.051 ( 0.064 0.165) 0.381 PCB209 0.040 ( 0.322 0.401) 0.829 Gender 0.028 ( 0.119 0.063) 0.545 Age 0.001 ( 0.006 0.004) 0.805 BMI 0.006 (0.001 0.011) 0.012 Smoking status 0.048 ( 0.065 0.162) 0.402 SumPCB 0.016 ( 0.163 0.131) 0.829 The model included gender, age, BMI, current smoking behavior, and one serum PCB level (lipid adjusted); FT4 was used as a square root transformed variable; All PCB levels were used after log transformation. Table S4. Linear regression for FT4 with hydroxylated PCB metabolites and total lipids. 4HO-PCB79 0.023 0.082 4HO-PCB120 0.022 0.083 3HO-PCB118 0.002 0.112 Gender 0.030 ( 0.120 0.061) 0.519 Age 0.001 ( 0.003 0.004) 0.766 BMI 0.008 (0.003 0.013) 0.002 Smoke 0.077 ( 0.039 0.193) 0.192 Total lipids 0.000 ( 0.001 0.000) 0.019 4HO-PCB79 0.028 ( 0.135 0.078) 0.602 Gender 0.030 ( 0.121 0.060) 0.507 Age 0.001 ( 0.003 0.005) 0.634 BMI 0.007 (0.003 0.012) 0.002 Smoke 0.072 ( 0.042 0.186) 0.216 Total lipids 0.000 ( 0.001 0.000) 0.018 4HO-PCB120 0.121 ( 0.507 0.266) 0.539 Gender 0.024 ( 0.113 0.065) 0.592 Age 0.001 ( 0.002 0.005) 0.442 BMI 0.005 (0.000 0.010) 0.061 Smoke 0.060 ( 0.053 0.173) 0.293 Total lipids 0.000 ( 0.001 0.000) 0.024 3HO-PCB118 0.147 ( 0.267 0.028) 0.016

S8 of S11 Table S4. Cont. 4HO-PCB107 0.021 0.083 3HO-PCB153 0.017 0.086 4HO-PCB146 0.015 0.088 4HO-PCB127 0.025 0.081 3HO-PCB138 0.016 0.087 Gender 0.029 ( 0.120 0.061) 0.520 Age 0.000 ( 0.005 0.004) 0.852 BMI 0.008 (0.003 0.013) 0.002 Smoke 0.065 ( 0.051 0.181) 0.268 Total lipids 0.000 ( 0.001 0.000) 0.024 4HO-PCB107 0.060 ( 0.109 0.229) 0.484 Gender 0.026 ( 0.117 0.064) 0.567 Age 0.001 ( 0.006 0.004) 0.593 BMI 0.009 (0.003 0.015) 0.002 Smoke 0.074 ( 0.041 0.188) 0.205 Total lipids 0.000 ( 0.001 0.000) 0.027 3HO-PCB153 0.0120 ( 0.115 0.355) 0.315 Gender 0.030 ( 0.120 0.060) 0.511 Age 0.002 ( 0.007 0.004) 0.489 BMI 0.009 (0.004 0.015) 0.001 Smoke 0.072 ( 0.042 0.186) 0.213 Total lipids 0.000 ( 0.001 0.000) 0.019 4HO PCB146 0.118 ( 0.086 0.323) 0.255 Gender 0.031 ( 0.121 0.060) 0.504 Age 0.001 ( 0.003 0.004) 0.740 BMI 0.007 (0.002 0.012) 0.004 Smoke 0.072 ( 0.042 0.187) 0.216 Total lipids 0.000 ( 0.001 0.000) 0.018 4HO-PCB127 0.040 ( 0.333 0.253) 0.787 Gender 0.025 ( 0.116 0.065) 0.584 Age 0.001 ( 0.007 0.004) 0.562 BMI 0.009 (0.004 0.014) 0.001 Smoke 0.074 ( 0.041 0.188) 0.205 Total lipids 0.000 ( 0.001 0.000) 0.025 3HO-PCB138 0.106 ( 0.089 0.302) 0.285

S9 of S11 Table S4. Cont. 4HO-PCB130 0.025 0.081 4HO-PCB163 0.014 0.089 4HO-PCB187 0.023 0.082 4HO-PCB162 0.024 0.082 4HO-PCB177 0.022 0.083 Gender 0.030 ( 0.120 0.061) 0.519 Age 0.000 ( 0.003 0.004) 0.811 BMI 0.008 (0.003 0.012) 0.002 Smoke 0.072 ( 0.042 0.187) 0.215 Total lipids 0.000 ( 0.001 0.000) 0.020 4HO-PCB130 0.044 ( 0.493 0.581) 0.871 Gender 0.032 ( 0.122 0.058) 0.485 Age 0.002 ( 0.006 0.003) 0.519 BMI 0.009 (0.004 0.014) 0.001 Smoke 0.072 ( 0.042 0.186) 0.216 Total lipids 0.000 ( 0.001 0.000) 0.023 4HO-PCB163 0.184 ( 0.109 0.478) 0.217 Gender 0.032 ( 0.123 0.059) 0.486 Age 0.000 ( 0.004 0.004) 0.935 BMI 0.008 (0.003 0.014) 0.003 Smoke 0.073 ( 0.042 0.187) 0.212 Total lipids 0.000 ( 0.001 0.000) 0.018 4HO-PCB187 0.057 ( 0.159 0.272) 0.603 Gender 0.031 ( 0.121 0.059) 0.499 Age 0.000 ( 0.005 0.004) 0.942 BMI 0.008 (0.003 0.013) 0.003 Smoke 0.067 ( 0.050 0.184) 0.262 Total lipids 0.000 ( 0.001 0.000) 0.023 4HO-PCB162 0.056 ( 0.199 0.312) 0.665 Gender 0.029 ( 0.119 0.061) 0.528 Age 0.000 ( 0.004 0.004) 0.993 BMI 0.008 (0.003 0.013) 0.002 Smoke 0.073 ( 0.041 0.188) 0.209 Total lipids 0.000 ( 0.001 0.000) 0.020 4HO-PCB177 0.091 ( 0.199 0.381) 0.536

S10 of S11 Table S4. Cont. 3HO-PCB180 0.003 0.110 4HO-PCB172 0.010 0.094 4HO-PCB193 0.002 0.112 4diMeO-PCB202 0.025 0.081 4HO-PCB208 0.020 0.084 Gender 0.041 ( 0.130 0.049) 0.381 Age 0.002 ( 0.007 0.002) 0.257 BMI 0.010 (0.005 0.015) 0.000 Smoke 0.083 ( 0.030 0.196) 0.148 Total lipids 0.000 ( 0.001 0.000) 0.020 3HO-PCB180 0.287 (0.047 0.527) 0.020 Gender 0.037 ( 0.127 0.053) 0.420 Age 0.003 ( 0.008 0.003) 0.334 BMI 0.010 (0.004 0.016) 0.001 Smoke 0.073 ( 0.040 0.187) 0.205 Total lipids 0.000 ( 0.001 0.000) 0.024 4HO-PCB172 0.203 ( 0.051 0.457) 0.116 Gender 0.034 ( 0.123 0.055) 0.447 Age 0.001 ( 0.005 0.003) 0.541 BMI 0.009 (0.004 0.014) 0.000 Smoke 0.071 ( 0.042 0.184) 0.215 Total lipids 0.000 ( 0.001 0.000) 0.025 4HO-PCB193 0.365 (0.071 0.658) 0.015 Gender 0.030 ( 0.120 0.061) 0.517 Age 0.001 ( 0.003 0.004) 0.796 BMI 0.008 (0.003 0.004) 0.002 Smoke 0.072 ( 0.043 0.187) 0.217 Total lipids 0.000 ( 0.001 0.000) 0.019 4diMeO-PCB202 0.007 ( 0.330 0.315) 0.964 Gender 0.029 ( 0.120 0.061) 0.521 Age 0.000 ( 0.003 0.004) 0.882 BMI 0.008 (0.003 0.013) 0.002 Smoke 0.073 ( 0.042 0.187) 0.211 Total lipids 0.000 ( 0.001 0.000) 0.018 4HO-PCB208 0.121 ( 0.169 0.410) 0.412

S11 of S11 Table S4. Cont. sum HO-PCBs 0.020 0.084 Gender 0.031 ( 0.121 0.059) 0.500 Age 0.001 ( 0.006 0.004) 0.722 BMI 0.009 (0.003 0.014) 0.003 Smoke 0.070 ( 0.045 0.184) 0.231 Total lipids 0.000 ( 0.001 0.000) 0.020 Sum HO-PCBs 0.091 ( 0.148 0.329) 0.455 2016 by the authors; licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons by Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).