Demographic description of KC patient groups.

<p>Demographic description of KC patient groups.</p

Comparison of microvascular remodeling in the urinary bladders of control and KC patients under electron microscopy.

<p>(A) Controls, BM (arrowheads): (i) small and (ii) greater vessels. (B) KC, duplication of BM (arrowheads): (i) small and (ii) greater vessels. (C) Bar graph of the increased thickness (μm) of BM in the KC (<i>n</i> = 36) and control (<i>n</i> = 7) groups: (i) small and (ii) greater vessels; obtained using the Mann–Whitney U test for statistical analysis. (A–C) Red bars: 2 μm.</p

Quantitative PLA assay of co-expression of mesenchymal transition markers in urinary bladder.

<p>(A, B, D) Red PLA signal dots represent the co-expressions of endothelial markers CD31 and mesenchymal marker FSP1 with close proximity on endothelial cells. Endothelial cells with circular arrangement (white arrows) and their surrounding PLA signal dots in the submucosa layer of the urinary bladder of the control (A; <i>n</i> = 9) and KC (B; <i>n</i> = 11) groups were further selected and counted. (D) Higher-magnification view over the microvessels of KC from panel (B); white bars: 50 μm. (C) Bar graph of PLA dots per endothelial cell count, used for comparison of the two groups (mean PLA dots per cell: KC, 2.32; control, 0.16); the Mann–Whitney U test was applied for statistical analysis.</p

Correlation between increased thickness of BM of vessels and clinical parameters, obtained using Spearman’s correlation coefficient for statistical analysis.

<p>Correlation between increased thickness of BM of vessels and clinical parameters, obtained using Spearman’s correlation coefficient for statistical analysis.</p

Demographic and cystoscopic findings in the control and KC patient groups.

<p>Demographic and cystoscopic findings in the control and KC patient groups.</p

Microvascular remodeling in the urinary bladder.

<p>(A, B) Histology of bladder biopsy of patient with KC reveals many eosinophils (black arrowheads) and vascular wall thickening (long black arrow) in suburothelial tissue (hematoxylin and eosin stain): (A) reduced from x 100; (B) reduced from x 400. (C) Small vessel from a normal control subject revealing an endothelial cell (long red arrow) with a single layer of BM (red arrowheads). (D) Small vessel from a KC patient revealing onion-skin duplication of the BM (red double-ended arrow); the increasing thickness of the BM was measured after division into four quarters, using two straight lines intersecting at the center of the vascular lumen. (E) Schematic representation of the duplication of the BM in greater (upper panel) and small (lower panel) vessels. (A, B) White bars: 50 μm; (C, D) red bars: 2 μm.</p

Immunohistochemial comparison of the expression of NMDAR1 on vessels and EndoMT markers in the urinary bladder.

<p>Biopsy specimens of frozen sections from the urinary bladders of (A, C, E) normal control subjects and (B, D, F) KC patients. (A, B) Double-immunofluorescence staining with antibody to endothelial marker CD31 (green) and NMDAR1 (red); DAPI was used as a nuclear stain (blue); co-expression of CD31 and NMDAR1 was evident in two groups (yellow, denoted by white arrows). (C–F) Double-immunofluorescence staining with antibody to endothelial marker CD31 (green) and two mesenchymal markers: FSP1 (red, in panels C and D) and α-SMA (red, in panels E and F); DAPI was used as a nuclear stain (blue); co-expression of CD31 and FSP1 or α-SMA was evident in three groups (yellow, denoted by white arrows) (white bars: 50 μm).</p

Duration free from UTS (years) of subjects with and without UI. (N = 10,662).

Duration free from UTS (years) of subjects with and without UI. (N = 10,662).</p

Characteristics of subjects (study group and control group B^a) with/without development of upper UTS. (N = 10,662).

Characteristics of subjects (study group and control group Ba) with/without development of upper UTS. (N = 10,662).</p

Association between UI and development of upper UTS (N = 10,662).

<p>Association between UI and development of upper UTS (N = 10,662).</p