Cytological and molecular features of the 25 cytology samples.

<p><b>Note:</b> NT = non-tumor; T = tumor; H = >10,000 cells; L = >5,000 and <10,000 cells.</p

Selection of the cytology specimens.

<p>Only cases with >5,000 well-preserved and well-visualized urothelial cells (as observed in 10 fields at 4x magnification) were selected. Cases largely constituted by red blood cells or inflammatory cells were excluded.</p

Total RNA extracted from archival urine cytology smears is suitable for miRNA expression profiling.

<p>Box plots show differences in miRNAs expression between non-tumor (NT) and low grade urothelial carcinoma samples (T). *<i>p</i><0.05.</p

RNA amount cut-off values for qRT-PCR analysis using archival cytology smears.

<p>RNA amount cut-off values for qRT-PCR analysis using archival cytology smears.</p

Identification of the minimal RNA quantity to obtain an adequate qRT-PCR reaction from cytology specimens.

<p>(<b>A</b>) The relationship between Ct and ldose for the two cell lines (BxPc3 and Capan-1) was analyzed as covariance (ANCOVA) model and the minimal RNA quantity was identified in terms of the estimated slope. For RNU6B, the ANCOVA analysis (R2 = 0.99) revealed that the slope of the relationship between ldose and Ct is the same in the two cells lines (p = 0.55) and it is equal to −3.73 (±0.07, p<0.0001), with efficiency <i>e</i> = 1.70. The estimated models are Ct = 27.65–3.55*ldose for BxPc3 and Ct = 24.97–3.55*ldose for Capan-1. (<b>B</b>) For miR-205 ANCOVA analysis (R2 = 0.98) revealed that the slope of the relationship between ldose and Ct is the same in the two cell lines and is equal to −4.00 (±0.12, p<0.000), with efficiency <i>e</i> = 1.50. The estimated model is Ct = 29.09–3.64*ldose both for BxPc3 and Capan-1. (<b>C</b>) To estimate the relationship between Ct and ldose on the 15 non-neoplastic patients, we used linear mixed effect models including a random effect for the subject. (<b>D</b>) The minimal RNA quantity was selected in terms of the fixed effect slope. For miR-205, when considering all the dilutions, the LME analysis (R2 = 0.92) revealed that the fixed effect slope of the relationship between ldose and Ct is −1.73 (±0.13, p<0.000), with efficiency <i>e</i> = 4.78.</p

Immunohistochemistry results.

<p>Overall, grade, stage, vascular and pleural invasion distribution of immunohistochemical staining scores for lumican in lung adenocarcinoma (<b>A</b>) and in the surrounding stroma (<b>B</b>).</p

Representative images from the considered series.

<p>In normal lung (<b>A and B</b>), lumican immunoreaction was detectable not only in the cytoplasm of pneumocytes, columnar cells of the bronchi, fibroblasts (<i>inset</i>), and vascular smooth muscle cells, but also in collagen fibers of the perivascular and peribronchial connective tissues. In lung adenocarcinoma (<b>C and D</b>), lumican immunostaining was low in tumor cells and more diffuse in the surrounding tumor stroma and extracellular space. (Original magnification 200x in A and D, 100x in B, and 40x in C).</p

Immunoblotting results.

<p>Immunoblotting analysis of lumican in Nutridoma and both cell lysate and supernatant of A549 lung adenocarcinoma cell line and fibroblast culture.</p

Proteomic analysis results.

<p>Proteins with different expression between lung adenocarcinoma and non-neoplastic pleural effusions.</p><p>Proteomic analysis results.</p

Representative images from the tumor cell-blocks.

<p>Lumican immunostaining was absent in lung adenocarcinoma cells (<b>A</b>), whereas it was strong and diffuse in breast carcinoma (invasive carcinoma of no special type) cells (<b>B</b>). (Original magnification 400x in A and B).</p