Angiogenesis and cancer invasiveness greatly contribute to cancer malignancy.Arf6 and its effector, AMAP1, are frequently overexpressed in breast cancer, and constitute a central pathway to induce the invasion and metastasis. In this pathway, Arf6 is activated by EGFR via GEP100. Arf6 is highly expressed also in human umbilical vein endothelial cells (HUVECs) and is implicated in angiogenesis. Here, we found that HUVECs also highly express AMAP1, and that vascular endothelial growth factor receptor-2 (VEGFR2) recruits GEP100 to activate Arf6. AMAP1 functions by binding to cortactin in cancer invasion and metastasis. We demonstrate that the same GEP100-Arf6-AMAP1-cortactin pathway is essential for angiogenesis activities, including cell migration and tubular formation, as well as for the enhancement of cell permeability and VE-cadherin endocytosis of VEGF-stimulated HUVECs. Components of this pathway are highly expressed in pathologic angiogenesis, and blocking of this pathway effectively inhibits VEGF- or tumor-induced angiogenesis and choroidal neovascularization. The GEP100-Arf6-AMAP1-cortactin pathway, activated by receptor tyrosine kinases, appears to be common in angiogenesis and cancer invasion and metastasis, and provides their new therapeutic targets.

PurposeThe aims of this study were to analyze optical coherence tomography (OCT) imaging of large macular holes (MHs) treated with inverted internal limiting membrane (ILM) flap technique and to perform a histological examination of an ILM-like membrane tissue obtained during vitrectomy.Patients and methodsThis is a retrospective observational case study. Nine patients, comprising of five males and four females, showing large and myopic MHs, underwent pars plana vitrectomy (PPV) with inverted ILM flap technique assisted by brilliant blue G (BBG) staining. Ophthalmological findings including visual acuity and OCT were investigated based on medical records. Formalin-fixed paraffin-embedded tissue section of an ILM-like membrane was submitted for immunohistochemistry with glial fibrillary acidic protein (GFAP).ResultsILM was clearly stained with BBG in eight patients, whereas the ILM in one case revealed no staining with BBG during PPV. Visual acuities improved to >0.2 LogMAR in six patients. The complete closure of MH following PPV with inverted ILM technique was eventually achieved in all patients determined by OCT imaging (100%). Only one patient showed recovery of ellipsoid zone and interdigitation zone following the surgery. Elongation of outer nuclear layer was noted in three eyes. The ILM-like membrane not stained with BBG histologically revealed an amorphous structure admixed with GFAP-positive mononuclear cell infiltration.ConclusionPPV with inverted ILM flap technique achieved 100% closure rates with favorable configuration at an initial surgery in large MHs. Our histopathological data also suggest that even BBG staining-negative membrane may be a useful material for autologous transplantation to the hole.

Our results indicate the pathological role of sVAP-1/SSAO to generate hydrogen peroxide and toxic aldehyde ACR, both of which are associated with oxidative stress, as a consequence of spermine oxidation in eyes with PDR.

1Purpose: Vascular endothelial growth factor C (VEGF-C) and its receptor 2 VEGFR-3 mediate lymphangiogenesis. In this study, we analyzed the 3 expression of VEGF-C and VEGFR-3 as well as lymphatic vessels in the 4 pterygium and normal conjunctiva of humans. 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 Most studies have supported the hypothesis that VEGF-C is of prognostic 9 value, and promotes lymphatic tumor progression in various human cancers, 10 including gastric, breast, colon, lung, head and neck, and ovarian 11 carcinomas. (Herrmann et al., 2007; Jüttner et al., 2006; Miyata et al., 2006; 12 Trojan et al., 2006) In the ophthalmology field, VEGF-C and its receptor, Dewaxed paraffin sections were immunostained using the complex method. 2Formalin-fixed, paraffin-embedded serial tissue sections were cut at a 4 µm 3 thickness and endogenous peroxidase activity was inhibited by immersing 4 the slides in 3% hydrogen peroxide in methanol for 10 min. As a 5 pretreatment, microwave-based antigen retrieval was performed in PBS. antibody (1:500 dilution; Invitrogen). All slides were examined using a 5Keyenece BZ-9000 (Keyence, Osaka, Japan) microscope. (1:500; described above) and anti-VEGFR-3 polyclonal antibody (1:500; Results 2LVD is high in pterygium 3 D2-40-positive lymphatic vessels were clearly seen in the stroma of the 4 pterygium and normal conjunctiva (Figure 1, arrows). The LVD of the 5 pterygium and normal conjunctiva was 8.2±2.8 and 4.8±0.3, respectively. 6The LVD was significantly higher in the pterygium than in the normal 7 conjunctiva (P < 0.001, Table 1). In contrast, the LVD/BVD ratio of the 8 pterygium (0.86±0.26) was not significant difference compared to that of 9 the normal conjunctiva (0.71±0.05) (P > 0.05, Table 1). 11 VEGF-C is expressed in pterygium 12The expression of VEGF-C in the pterygium and normal conjunctiva was Figure 4A, B). The 6 number of VEGF-A-immunopositive cells (54%) was significantly higher 7 than that of VEGF-C (44%) (n=9, P < 0.05, Table 3). Immunoreactivity for p<0.05, Figure 6A ). VEGF-C is secreted in serum-starved cultured conjunctival epithelial VEGF-C promotes lymphatic endothelial cell proliferation 9To investigate the role of VEGF-C in pterygium pathogenesis, we studied at 24 hours (n=4, p<0.05, Figure 6B). In this study, we demonstrated that VEGF-C was expressed in pterygial 1 tissues using Western blot and imm...

Among spontaneous neoplasms of the rat central nervous system, the discrimination between astrocytoma and malignant reticulosis (MR) is sometimes difficult because of their similar cell morphology and infiltration patterns. In the present study, we carried out histological and immunohistochemical analyses on a total of sixty-four cases in Sprague-Dawley and F344 rats. These cases were diagnosed as benign/malignant astrocytoma containing no neoplastic oligodendroglial elements or MR according to the diagnostic criteria of the World Health Organization International Classification of Rodent Tumors (Mohr et al. 1994). Astrocytomas were divided into three types and MR into two types based on the number of lesions, cellularity and infiltration patterns, and so on. Although the neoplastic cells from all types showed various immunoreactivities for RM-4 (anti-rat macrophages and dendritic cells), ED-1, and/or vimentin, there were no distinctive differences among these types, and most cells that were positive for RM-4 were also positive for ED-1. None of the tumor types showed any reactivity for GFAP or S-100 protein. From the results of morphological and immunohistochemical examinations, it was indicated that there are no distinctive differences between spontaneous astrocytomas and MR in rats, and they are probably derived from the same cell lineage, that is, microglia, macrophage, or radial glia.