EPDR1 is a noncanonical effector of insulin-mediated angiogenesis regulated by an endothelial-specific TGF-ß receptor complex.

Insulin signaling in blood vessels primarily functions to stimulate angiogenesis and maintain vascular homeostasis through the canonical PI3K and MAPK signaling pathways. However, angiogenesis is a complex process coordinated by multiple other signaling events. Here, we report a distinct crosstalk between the insulin receptor and endoglin/activin receptor-like kinase 1 (ALK1), an endothelial cell-specific TGF-ß receptor complex essential for angiogenesis. While the endoglin-ALK1 complex normally binds to TGF-ß or bone morphogenetic protein 9 (BMP9) to promote gene regulation via transcription factors Smad1/5, we show that insulin drives insulin receptor oligomerization with endoglin-ALK1 at the cell surface to trigger rapid Smad1/5 activation. Through quantitative proteomic analysis, we identify ependymin-related protein 1 (EPDR1) as a major Smad1/5 gene target induced by insulin but not by TGF-ß or BMP9. We found endothelial EPDR1 expression is minimal at the basal state but is markedly enhanced upon prolonged insulin treatment to promote cell migration and formation of capillary tubules. Conversely, we demonstrate EPDR1 depletion strongly abrogates these angiogenic effects, indicating that EPDR1 is a crucial mediator of insulin-induced angiogenesis. Taken together, these results suggest important therapeutic implications for EPDR1 and the TGF-ß pathways in pathologic angiogenesis during hyperinsulinemia and insulin resistance.

Anatomical evaluation of CT-MRI combined femoral model.

BACKGROUND: Both CT and MRI are complementary to each other in that CT can produce a distinct contour of bones, and MRI can show the shape of both ligaments and bones. It will be ideal to build a CT-MRI combined model to take advantage of complementary information of each modality. This study evaluated the accuracy of the combined femoral model in terms of anatomical inspection. METHODS: Six normal porcine femora (180 +/- 10 days, 3 lefts and 3 rights) with ball markers were scanned by CT and MRI. The 3D/3D registration was performed by two methods, i.e. the landmark-based 3 points-to-3 points and the surface matching using the iterative closest point (ICP) algorithm. The matching accuracy of the combined model was evaluated with statistical global deviation and locally measure anatomical contour-based deviation. Statistical analysis to assess any significant difference between accuracies of those two methods was performed using univariate repeated measures ANOVA with the Turkey post hoc test. RESULTS: This study revealed that the local 2D contour-based measurement of matching deviation was 0.5 +/- 0.3 mm in the femoral condyle, and in the middle femoral shaft. The global 3D contour matching deviation of the landmark-based matching was 1.1 +/- 0.3 mm, but local 2D contour deviation through anatomical inspection was much larger as much as 3.0 +/- 1.8 mm. CONCLUSION: Even with human-factor derived errors accumulated from segmentation of MRI images, and limited image quality, the matching accuracy of CT-&-MRI combined 3D models was 0.5 +/- 0.3 mm in terms of local anatomical inspection.

Synthesis and evaluation of 3-aminopropionyl substituted fentanyl analogues for opioid activity.

An enkephalin analogue coupled to 'aminofentanyl' has been synthesized and tested for biological activities at the mu and delta opioid receptors. Aminofentanyl which represents a structural derivative of fentanyl has been synthesized by acylation of 1-(2-phenethyl)-4-(N-anilino)piperidine with phthaloyl protected beta-alaninyl chloride in the presence of DIPEA, followed by deprotection with hydrazine hydrate. Aminofentanyl has also been successfully acylated with ethyl isocyanate, various acid anhydrides, to further investigate structure-activity relationships of these new fentanyl derivatives. Among the new derivatives compound 7 which carries a Tyr-D-Ala-Gly-Phe opioid message sequence showed good opioid affinity (1 nM at both delta and mu opioid receptors) and bioactivity (34.9 nM in MVD and 42 nM in GPI/LMMP bioassays).