Fig. 1 Changes in biochemical assessment of CKD-MBD over time: Cy/+ rats were treated with diet only (CKD CTL), R-568, R-568 + Ca or calcium alone (Ca).

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Fig. 1 Changes in biochemical assessment of CKD-MBD over time: Cy/+ rats were treated with diet only (CKD CTL), R-568, R-568 + Ca or calcium alone (Ca). (A) PTH increased over time in the control animals but was equally suppressed by all three treatments (P < 0.0001). (B) The calcium level was increased in the Ca-treated animals and decreased in the R-568-treated animals (each P < 0.001). In contrast, the addition of calcium to R-568 normalized the calcium to that of the control animals. (C) Phosphorus levels decreased very slightly in the control animals until after 34 weeks when there was an increase with further decline in renal function and increase in PTH. The animals treated with R-568 + Ca or Ca alone had a significant decrease in phosphorus compared to control at 38 weeks (P < 0.001). In contrast, the R-568-treated animals had an increase in the plasma phosphorus at 29 and 34 weeks compared to control, with a return to control levels at 38 weeks (P < 0.001). Solid black bars = CKD animals without treatment (control); open white bars = CKD given R-568; hatched bars = CKD animals given R-568 plus 2% calcium gluconate in the drinking water; grey bars = CKD animals given only 2% calcium gluconate in the drinking water. The number of animals at each time point was 31–33 at 20 weeks, 15–16 at 29 weeks, 29–32 at 34 weeks and 15–16 at 38 weeks. From: R-568 reduces ectopic calcification in a rat model of chronic kidney disease-mineral bone disorder (CKD-MBD) Nephrol Dial Transplant. 2009;24(8):2371-2377. doi:10.1093/ndt/gfp078 Nephrol Dial Transplant | Published by Oxford University Press on behalf of ERA-EDTA [2009]. All rights reserved. For Permissions, please e-mail: journals.permissions@oxfordjournals.orgOxford University Press

Fig. 2 Thoracic aorta calcification Fig. 2 Thoracic aorta calcification. At the time of sacrifice, the thoracic aorta was removed and its calcium and phosphorus content determined biochemically. At 34 weeks, there was no difference in the calcium content (A), but the phosphorus content (B) was greater in the calcium-treated animals than all of the other groups (P = 0.002). At 38 weeks, the calcium-treated animals had greater calcium (P = 0.04; A) and phosphorus content (P = 0.03; B) compared with the R-568-treated animals. At 38 weeks, animals treated with R-568 + Ca had higher calcium content than R-568 alone. Solid black bars = CKD animals without treatment (control); open white bars = CKD given R-568; hatched bars = CKD animals given R-568 plus 2% calcium gluconate in the drinking water; grey bars = CKD animals given only 2% calcium gluconate in the drinking water. From: R-568 reduces ectopic calcification in a rat model of chronic kidney disease-mineral bone disorder (CKD-MBD) Nephrol Dial Transplant. 2009;24(8):2371-2377. doi:10.1093/ndt/gfp078 Nephrol Dial Transplant | Published by Oxford University Press on behalf of ERA-EDTA [2009]. All rights reserved. For Permissions, please e-mail: journals.permissions@oxfordjournals.orgOxford University Press

Fig. 3 Aortic arch calcification Fig. 3 Aortic arch calcification. The aortic arch was dissected from above the aortic valve to the ligamentum arteriosum from either 34- or 38-week animals and its calcium (A) and phosphorus (B) content analysed biochemically (n = 6–8 per group). The results are expressed as μmol/g of tissue. At 34 and 38 weeks (A), there was a trend for a reduction in calcium content in R-568-treated animals compared with calcium alone, but this did not reach statistical significance. At 34 weeks (B), there was no difference in the phosphorus content. In contrast, at 38 weeks, there was increased phosphorus content in the rats treated with calcium alone compared with the other treatment groups (P = 0.004). Solid black bars = CKD animals without treatment (control); open white bars = CKD given R-568; hatched bars = CKD animals given R-568 plus 2% calcium gluconate in the drinking water; grey bars = CKD animals given only 2% calcium gluconate in the drinking water. From: R-568 reduces ectopic calcification in a rat model of chronic kidney disease-mineral bone disorder (CKD-MBD) Nephrol Dial Transplant. 2009;24(8):2371-2377. doi:10.1093/ndt/gfp078 Nephrol Dial Transplant | Published by Oxford University Press on behalf of ERA-EDTA [2009]. All rights reserved. For Permissions, please e-mail: journals.permissions@oxfordjournals.orgOxford University Press

Fig. 4 Heart calcification Fig. 4 Heart calcification. The upper atrium/aortic valve (A) and left ventricle (B) from either 34- or 38-week animals was isolated and the calcium content analysed biochemically (n = 6–8 per group). The results are expressed as μmol/g of tissue. At 34 weeks for the aortic valve (A), there was a trend towards less calcium (P = 0.053) in the R-568 alone treated animals, and by 38 weeks, both calcium-treated groups had more calcium content than the R-568-treated animals and controls (P = 0.035). The left ventricle (B) had less calcium at 34 weeks (P = 0.008) and 38 weeks (P = 0.005) in the R-568 alone treated animals than in the other treatment groups. Solid black bars = CKD animals without treatment (control); open white bars = CKD given R-568; hatched bars = CKD animals given R-568 plus 2% calcium gluconate in the drinking water; grey bars = CKD animals given only 2% calcium gluconate in the drinking water. From: R-568 reduces ectopic calcification in a rat model of chronic kidney disease-mineral bone disorder (CKD-MBD) Nephrol Dial Transplant. 2009;24(8):2371-2377. doi:10.1093/ndt/gfp078 Nephrol Dial Transplant | Published by Oxford University Press on behalf of ERA-EDTA [2009]. All rights reserved. For Permissions, please e-mail: journals.permissions@oxfordjournals.orgOxford University Press