The number of immunomarked MCs for each antibody

was separately determined for each group in histologic fields of higher density. Two previously calibrated and independent observers performed MC counting in 10 fields of control cases (5 fields at the epithelium–connective tissue junction and 5 in the reticular lamina propria), in 10 fields of ACs (5 at the epithelium–connective tissue junction and 5 in areas of solar elastosis), and in 15 fields of SCCs (5 at the epithelium–connective tissue junction, 5 in the tumor parenchyma, and 5 in the peritumoral stroma), at ×400 magnification (counting field area 0.20 mm2). MMP-9 immunohistochemical staining was evaluated through descriptive and semiquantitative Ku-0059436 clinical trial analysis. In the latter, we used scores adjusted from Franchi et al.25 for analysis of the epithelial tissue in ACs and control samples and of tumor cells in SCCs, based on the percentage of immunoreactive cells and their staining intensity Cyclopamine (Table II). The analysis was performed using a light microscope by 2 previously calibrated and independent observers. MC density descriptive analysis was expressed as confidence intervals (CIs) of the number of observations per mm2. Comparative analysis of means between groups and between histologic fields was performed using parametric 1-way analysis of variance (ANOVA). Paired multiple comparisons were then performed by using the Tukey test. MC migration was expressed as the ratio between c-Kit+

and tryptase+ densities, and the comparative analysis between groups was performed with 1-way anova and Tukey post hoc tests. Association between MC density and MMP-9 expression was assessed by using the Student t test. Differences were considered to be statistically significant when P < .05. The main histologic findings regarding the 20 specimens of ACs are summarized

in Table III. In the SCC analysis, 7 cases were classified as well differentiated, 3 as moderately differentiated, and 10 as undifferentiated. Resident aminophylline MCs were identified by the use of antitryptase antibody, and MC migration was evaluated with an anti–c-Kit antibody. Analysis of material submitted to immunohistochemistry showed that tryptase+ MCs were more strongly expressed in SCCs than in ACs and control samples (P < .001 [Tukey post hoc test]; Table IV). In these tumors, a significantly high density of tryptase+ MCs was found in the tumor stroma, surrounding the invasive epithelial nests and cords ( Fig. 1, a). A high expression of these cells was also observed near the lining epithelium adjacent to the tumor, though less than in the stroma (P = .007 [Tukey]). Moreover, a sparse density of these cells was observed in the lesion parenchyma compared with the tumor stroma (P < .001 [Tukey]). Regarding the immunostaining for c-Kit, we found a higher concentration of c-Kit+ MCs in SCCs than in ACs and control samples (P < .001 [Tukey]; Table IV), similar to what was found for tryptase.