Volume 69, Issue 12, Pages (June 2006)

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Volume 69, Issue 12, Pages 2227-2235 (June 2006) Renal crystal deposits and histopathology in patients with cystine stones  A.P. Evan, F.L. Coe, J.E. Lingeman, Y. Shao, B.R. Matlaga, S.C. Kim, S.B. Bledsoe, A.J. Sommer, M. Grynpas, C.L. Phillips, E.M. Worcester  Kidney International  Volume 69, Issue 12, Pages 2227-2235 (June 2006) DOI: 10.1038/sj.ki.5000268 Copyright © 2006 International Society of Nephrology Terms and Conditions

Figure 1 Endoscopic views of papilla from cystine stone patients. Papillary morphology varies from (a) normal in patient 1 to (arrows) flattened with greatly enlarged openings of the (b and d) BD. (c) Some dilated BD contained protruding plugs of (double arrow) crystalline material. (b, arrowhead) Small sites of suburothelial white plaque, termed Randall's plaque, were noted. Kidney International 2006 69, 2227-2235DOI: (10.1038/sj.ki.5000268) Copyright © 2006 International Society of Nephrology Terms and Conditions

Figure 2 Endoscopic unroofing of intra-luminal cystine deposits. (a–d, arrow) In two patients, large masses of crystalline material were seen at the time of PNL to lie under the urothelium at a site marked by a dilated BD. (b, arrowheads) Upon unroofing the urothelium during the biopsy procedure, (c, double arrow) deposits were exposed that were located within a tubular lumen. (d) Lumen of the (asterisk) BD after the deposit had been removed. Kidney International 2006 69, 2227-2235DOI: (10.1038/sj.ki.5000268) Copyright © 2006 International Society of Nephrology Terms and Conditions

Figure 3 Histological images of papillary biopsies from cystine stone formers. The papillary histopathology of the cystine patients varied from (a) normal to (b–d) regions of plugging, dilation, and injury of IMCD. Intra-luminal plugging with crystals was noted in (b–d, arrows) thin loops of Henle and (d, double arrows) IMCD. (b and c, asterisks) Dilated IMCDs without crystalline material were commonly seen. Original magnification, (a) × 700, (b) × 600,(c) × 800, and (d) × 550. Kidney International 2006 69, 2227-2235DOI: (10.1038/sj.ki.5000268) Copyright © 2006 International Society of Nephrology Terms and Conditions

Figure 4 Sites of crystalline deposition in loops of Henle and BD. (a, arrows) The initial site of crystallization in the thin loops of Henle appears to be at the apical cell surface. (a, double arrow) A site of complete plugging of the thin loop is seen. (b and c, arrowheads) Intra-luminal plugging also occurs in the BD where occasionally we observed layers of epithelial cells that are seen surrounding masses of crystals. Examination of crystalline material in the lumen of the BD (b) without and (c) with polarization microscopy suggests that the deposits are cystine and not apatite. Original magnification, (a) × 900, (b) × 1100, and (c) × 1100. Kidney International 2006 69, 2227-2235DOI: (10.1038/sj.ki.5000268) Copyright © 2006 International Society of Nephrology Terms and Conditions

Figure 5 Basement membrane and interstitial changes in cystine papillary biopsies. (a–c, arrows, magnified inset to b) Yasue staining for calcium detected tiny crystalline deposits in the basement membranes of thin loops of Henle, a pattern identical to the interstitial plaque termed Randall's type I plaque. (b and c, double arrows) Additional interstitial changes included varying degrees of interstitial fibrosis surrounding sites of intra-tubular crystalline deposits in either (b, asterisk) loops of Henle or (c, asterisk) IMCD. Original magnification, (a) × 900, (b) × 1200, and (c) and × 1100. Kidney International 2006 69, 2227-2235DOI: (10.1038/sj.ki.5000268) Copyright © 2006 International Society of Nephrology Terms and Conditions

Figure 6 Transmission electron microscopic images of cellular injury to thin loops of Henle. Epithelial cell injury ranged from an unusual amount of cellular debris in the (a, arrows) tubular lumen to frank necrosis exposing the (b, double arrow) tubular basement membrane. (b and c, asterisks) Crystalline material was seen admixed with cellular debris within the tubular lumen. Note the presence of crystalline deposits within the (a–c, arrowheads) basement membranes and in the interstitial space. Original magnification, (a) × 5000, (b) × 6000, and (c) × 5500. Kidney International 2006 69, 2227-2235DOI: (10.1038/sj.ki.5000268) Copyright © 2006 International Society of Nephrology Terms and Conditions

Figure 7 Endoscopic and μCT images of papillary tissue with enlarged BD. (a) An endoscopic view of a papilla with an enlarged opening of a BD with a protruding plug of crystalline deposit (arrow). A papillary biopsy was obtained that included the dilated BD. (b) A three-dimensional reconstructed μCT image of that papillary biopsy and the arrow indicates the site of the protruding plug seen in (a). The large, dense triangular deposit was determined to have an attenuation value of 8500, a value consistent with cystine. Surrounding this large deposit are several smaller regions of (double arrow) mineral that all had an attenuation value of 22 000, a value consistent with apatite. A single μCT slice of the biopsy is seen in (c) where a portion of the (arrow) large and (double arrows) small deposits are clearly seen. This biopsy was divided in half. The upper half containing the large crystalline deposit was used for μ-Fourier transform infrared microspectroscopy analysis, whereas the lower half was embedded in paraffin for histopathogy. (d) A paraffin section from the lower half of the biopsy showing Yasue-positive material in a (double arrow) thin loop of Henle. Original magnification, (d) × 900. Kidney International 2006 69, 2227-2235DOI: (10.1038/sj.ki.5000268) Copyright © 2006 International Society of Nephrology Terms and Conditions

Figure 8 μ-Fourier transform infrared microspectroscopy spectra of crystal deposits in cystine stone former. This figure illustrates a series of infrared spectra obtained for a set of standards (calcium oxalate, cystine, and hydroxyapatite). The spectrum for intra-luminal crystals (1), obtained from a region of deposit collected from the upper half of the biopsy seen in Figure 7, matched that of cystine. The spectrum obtained from intra-luminal crystals (2) in the thin limb of Henle's loop matched that of apatite, whereas intra-luminal crystals (2) was from a inner medullary collecting also matched with apatite. Kidney International 2006 69, 2227-2235DOI: (10.1038/sj.ki.5000268) Copyright © 2006 International Society of Nephrology Terms and Conditions

Figure 9 TEM electron diffraction pattern of intra-luminal deposit in a cystine stone former. An area of intra-tubular crystal deposition (thin loop of Henle) from Case 3 was analyzed by electron diffraction and identified as biologic hydroxyapatite. The pattern shows the sharp 002 diffraction peak reflection, the elongated nature of the crystals, and the broad diffraction band containing the 211, 112, and 300 diffraction peaks, which is a characteristic of biologic apatite. Kidney International 2006 69, 2227-2235DOI: (10.1038/sj.ki.5000268) Copyright © 2006 International Society of Nephrology Terms and Conditions

Figure 10 Histopathology of cortical biopsies from cystine stone formers. Cortical biopsies from the cystine SF showed no changes (a) in patient 1 to a (b) moderate degree of glomerular obsolescence, tubular atrophy, and interstitial fibrosis . In addition, we observed Yasue-positive deposits in the basement membranes of the (c and d, arrows) parietal epithelial cells and (d, double arrows) distal convoluted tubules. Original magnification, (a) × 700, (b) × 700, (c) × 1100, and (d) × 1100. Kidney International 2006 69, 2227-2235DOI: (10.1038/sj.ki.5000268) Copyright © 2006 International Society of Nephrology Terms and Conditions