Holistic approach to visualize and quantify collagen organization at macro, micro, and nano-scale.

BACKGROUND: There is scarcity of imaging and image processing techniques for accurate discrimination and quantitation of the dermal extracellular matrix (ECM), primarily collagen. The aim of this study was to develop and demonstrate a holistic imaging and image processing approach to visualize and quantify collagen remodeling at the macro-, micro- and nano-scale using histochemical imaging, Reflectance Confocal Microscopy (RCM), and Atomic Force Microscopy (AFM), respectively. MATERIAL AND METHODS: For proof-of-concept, a commercial anti-aging product known to induce collagen neo-synthesis and re-organization was tested ex vivo on human skin biopsies from two aged females. RESULTS: Relative to untreated skin, collagen fibers (RCM) and fibrils (AFM) were longer and aligned after treatment. The content of collagen and elastin (histochemical imaging and ELISA) statistically improved after treatment. CONCLUSION: Based on our findings, we can conclude: (1) AFM, RCM, and histochemical imaging can accurately discriminate collagen from other ECM components in the skin and (2) the image processing methods can enable quantitation and hence capture small improvements in collagen remodeling after treatment (commercial cosmetic product with collagen organizer technology as proof-of-concept). The reported holistic imaging approach has direct clinical implications for scientists and dermatologists to make quick, real-time, and accurate decisions in skin research and diagnostics.

pAKT Expression and Response to Sorafenib in Differentiated Thyroid Cancer.

Sorafenib has an antitumor activity in patients with radioactive iodine-refractory differentiated thyroid carcinoma (RAIR-DTC). Prior research has implicated signaling through the MAPK and AKT/PI3K pathways in the progression of DTC. To assess whether the activity of these pathways is predictive of response to sorafenib, we retrospectively studied molecular tumor markers from these two pathways from a phase 2 study of sorafenib in RAIR-DTC. Tumor samples from 40 of 53 DTC subjects obtained prior to initiation of sorafenib were immunostained with DAB-labeled antibodies to phospho-AKT (pAKT), phospho-ERK (pERK), and phospho-S6 (pS6). BRAFV600E genetic mutation analysis was performed on all samples. Expression levels and mutational status were compared to response and progression-free survival (PFS) for each patient. Low tumor expression of nuclear pAKT was associated with partial response to sorafenib (p < 0.01). Patients with nuclear pAKT expression that was below the median for our sample were more than three times as likely to have a partial response as patients with equal to or above median expression. There was no correlation between tumor expression of nuclear pERK or pS6 and response. Endothelial cell and pericyte expression of pERK, pAKT, and pS6 were not predictive of response. There was no correlation between BRAFV600E mutation status and partial response. No correlation was observed between either the expression of pAKT, pERK, or pS6, or the presence of the BRAFV600E mutation, and PFS. In conclusion, lower tumor expression of nuclear pAKT was associated with higher rate of response to sorafenib. This observation justifies evaluation of combination therapy with sorafenib and an inhibitor of the PI3K/AKT signaling pathway in RAIR-DTC.