1. Smooth muscle–specific actin levels in the urine of renal transplant recipients: Correlation with cyclosporine or tacrolimus nephrotoxicity 
Mark Haas, MD, PhD, Shane M. Meehan, MB, BCh, Michelle A. Josephson, MD, Ernst-Jan C. Wit, PhD, E.Steve Woodle, MD, J.Richard Thistlethwaite, MD, PhD 
American Journal of Kidney Diseases 
Volume 34, Issue 1, Pages (July 1999)
DOI: /S (99)
Copyright © 1999 National Kidney Foundation, Inc Terms and Conditions

2. Fig 1
Histological lesions of CSA or tacrolimus nephrotoxicity on renal biopsy. (A) Hyaline arteriolopathy (periodic acid–Schiff stain; original magnification ×200). (B) Thrombotic microangiopathy (hematoxylin-eosin stain; original magnification ×200). (C) Isometric tubular epithelial cell vaculolization (hematoxylin-eosin stain; original magnification ×100).
American Journal of Kidney Diseases  , 69-84DOI: ( /S (99) )
Copyright © 1999 National Kidney Foundation, Inc Terms and Conditions

3. Fig 2
(A) Western blot and (B) Coomassie blue–stained SDS-polyacrylamide gel of proteins of homogenized aortic smooth muscle preparation used for ELISA standards (lane 2 of each gel), purified bovine skeletal muscle actin (lane 3), and purified human nonmuscle (platelet) actin (lane 4). On each gel, lane 1 contained molecular mass standards (Bio-Rad, Hercules, CA) corresponding to the kilodalton values noted to the left of each gel (TD = tracking dye). The amount of protein added to each gel was approximately 66 μg for lane 2 (corresponding to approximately 12 μg of actin), 15 μg for lane 3, and 12 μg for lane 4. Western blotting was performed with the same anti-SMA antibody used in the ELISA assay, described in Materials and Methods.
American Journal of Kidney Diseases  , 69-84DOI: ( /S (99) )
Copyright © 1999 National Kidney Foundation, Inc Terms and Conditions

4. Fig 2
(A) Western blot and (B) Coomassie blue–stained SDS-polyacrylamide gel of proteins of homogenized aortic smooth muscle preparation used for ELISA standards (lane 2 of each gel), purified bovine skeletal muscle actin (lane 3), and purified human nonmuscle (platelet) actin (lane 4). On each gel, lane 1 contained molecular mass standards (Bio-Rad, Hercules, CA) corresponding to the kilodalton values noted to the left of each gel (TD = tracking dye). The amount of protein added to each gel was approximately 66 μg for lane 2 (corresponding to approximately 12 μg of actin), 15 μg for lane 3, and 12 μg for lane 4. Western blotting was performed with the same anti-SMA antibody used in the ELISA assay, described in Materials and Methods.
American Journal of Kidney Diseases  , 69-84DOI: ( /S (99) )
Copyright © 1999 National Kidney Foundation, Inc Terms and Conditions

5. Fig 3
Indirect immunofluorescence staining of human renal cortex with the anti-SMA antibody. (A) Phase-contrast image of the tissue; (B) fluorescence image of the same field. (A) The lumen of a small artery is indicated by A and the lumens of two arterioles by a. (B) Fluorescence is limited to the muscle layers of these three blood vessels. Similar findings were noted in other areas of the tissue sample ([A, B] original magnification ×100).
American Journal of Kidney Diseases  , 69-84DOI: ( /S (99) )
Copyright © 1999 National Kidney Foundation, Inc Terms and Conditions

6. Fig 4
Urine SMA levels in patients with and without evidence of CSA or tacrolimus nephrotoxicity on renal biopsy and in six healthy control subjects. (○), A single urine sample; (•), mean values; error bars, 1 SD. For samples corresponding to biopsies with no evidence of nephrotoxicity, data are shown for all such cases (All; n = 49); for cases with acute rejection on the biopsy (including Banff borderline lesions; n = 31), and for those without acute rejection. Other diagnoses included acute tubular necrosis, hypertensive nephrosclerosis, chronic rejection, nonspecific chronic changes, acute pyelonephritis, viral-induced interstitial nephritis, recurrent focal-segmental glomerulosclerosis, posttransplant lymphoproliferative disease, suspected obstructive uropathy, and no histologic abnormality; n = 18. Breakdown of patients with CSA or tacrolimus toxicity is shown in Fig 5. For mean values, a = significantly different (P < 0.05) from patients with CSA or tacrolimus nephrotoxicity by t -test and Wilcoxon's rank-sum test; b = significantly different (P < 0.01) from patients without CSA or tacrolimus nephrotoxicity (All) by t -test and Wilcoxon's rank-sum test.
American Journal of Kidney Diseases  , 69-84DOI: ( /S (99) )
Copyright © 1999 National Kidney Foundation, Inc Terms and Conditions

7. Fig 5
Urine SMA levels in patients with different histopathologic lesions of CSA or tacrolimus nephrotoxicity on renal biopsy. (○), A single urine sample; (•), mean values; error bars, 1 SD. Categories include all urine samples corresponding to a biopsy with evidence of toxicity (All; n = 40), arteriolar lesions (n = 28), isometric tubular epithelial cell vacuolization but no arteriolopathy (tubular only; n = 12), thrombotic microangiopathy (n = 6), and no histopathologic evidence of CSA or tacrolimus toxicity before the current biopsy (first diagnosis; n = 17, including 7 patients with tubular toxicity only). For mean values, a = significantly different (P < 0.05) from patients without CSA or tacrolimus nephrotoxicity by t -test and Wilcoxon's rank-sum test; b = significantly different (P < 0.05) from patients with CSA or tacrolimus nephrotoxicity (All) by t -test and Wilcoxon's rank-sum test; c = significantly different (P = 0.04) from patients with CSA or tacrolimus nephrotoxicity (All) by Wilcoxon's rank-sum test and P = by t -test; d = significantly different (P < 0.05) from patients with tubular toxicity only by t -test and Wilcoxon's rank-sum test.
American Journal of Kidney Diseases  , 69-84DOI: ( /S (99) )
Copyright © 1999 National Kidney Foundation, Inc Terms and Conditions

8. Fig 6
Correlation of urinary SMA levels with severity and location of arteriolar lesions on the corresponding renal biopsy. Mild, moderate, and severe arteriolopathy are defined in Materials and Methods. (○), A single urine sample; (•), mean values; error bars, 1 SD. By ANOVA, mean SMA levels were significantly different between the three categories of severity (P < 0.01). Mean SMA levels in patients with mild arteriolopathy were significantly different from those in patients with moderate arteriolopathy (P < 0.05) and severe arteriolopathy (P < 0.02) by both t -test and Wilcoxon's rank-sum test. Mean SMA levels in patients with moderate and severe arteriolopathy were significantly different by t -test (P = 0.03) but not by Wilcoxon's rank-sum test (P = 0.10).
American Journal of Kidney Diseases  , 69-84DOI: ( /S (99) )
Copyright © 1999 National Kidney Foundation, Inc Terms and Conditions

9. Fig 7
Correlation of urinary SMA levels with whole-blood levels of (A) CSA (n = 21) and (B) tacrolimus (n = 67) on the day of the biopsy and urine sample. Efforts were routinely made to collect blood samples at a time corresponding to that of an expected trough level, based on the time of the previous CSA or tacrolimus dose. (○), A urine sample for which the corresponding biopsy showed no evidence of CSA or tacrolimus nephrotoxicity; (•), urine samples corresponding to biopsies with evidence of toxicity. For both A and B, data for all patients and for only those with biopsy evidence of toxicity were fit by linear regression analysis, with r as the correlation coefficient. For CSA, there was a significant correlation between urine SMA and blood CSA levels for all patients (r2 = 0.21; P = 0.037) and for the six patients with CSA nephrotoxicity (r2 = 0.81; P = 0.014). There was no significant correlation between urine SMA and blood tacrolimus levels (r2 = for all patients and r2 = for the 34 patients with tacrolimus nephrotoxicity; P > 0.60 in each instance).
American Journal of Kidney Diseases  , 69-84DOI: ( /S (99) )
Copyright © 1999 National Kidney Foundation, Inc Terms and Conditions

10. Fig 7
Correlation of urinary SMA levels with whole-blood levels of (A) CSA (n = 21) and (B) tacrolimus (n = 67) on the day of the biopsy and urine sample. Efforts were routinely made to collect blood samples at a time corresponding to that of an expected trough level, based on the time of the previous CSA or tacrolimus dose. (○), A urine sample for which the corresponding biopsy showed no evidence of CSA or tacrolimus nephrotoxicity; (•), urine samples corresponding to biopsies with evidence of toxicity. For both A and B, data for all patients and for only those with biopsy evidence of toxicity were fit by linear regression analysis, with r as the correlation coefficient. For CSA, there was a significant correlation between urine SMA and blood CSA levels for all patients (r2 = 0.21; P = 0.037) and for the six patients with CSA nephrotoxicity (r2 = 0.81; P = 0.014). There was no significant correlation between urine SMA and blood tacrolimus levels (r2 = for all patients and r2 = for the 34 patients with tacrolimus nephrotoxicity; P > 0.60 in each instance).
American Journal of Kidney Diseases  , 69-84DOI: ( /S (99) )
Copyright © 1999 National Kidney Foundation, Inc Terms and Conditions