Key Words: ADAMs Ⅲ metalloproteinase-disintegrins Ⅲ TNF␣-convertase Ⅲ proliferative retinopathy Ⅲ pathological neovascularization P athological neovascularization has a critical role in diseases such as cancer, 1,2 rheumatoid arthritis 3 and proliferative retinopathies, including retinopathy of prematurity, diabetic retinopathy and the wet form of macular degeneration. 4,5 Therefore molecules with roles in pathological neovascularization are considered potential targets for treatment of these conditions. Previous studies have identified a role for the cell surface metalloproteinase ADAM17 (a disintegrin and metalloproteinase 17, also referred to as TACE [tumor necrosis factor ␣-converting enzyme]) in crosstalk between the VEGFR2 and extracellular signal-regulated kinase 1/2 in endothelial cells, and in processing several receptors with key functions in angiogenesis, including the VEGFR2 and Tie2. 6 The goal of the present study was to determine whether ADAM17 has a role in angiogenesis or pathological neovascularization in vivo by subjecting conditional knockout mice carrying floxed alleles of ADAM17 7 and a Cre-recombinase expressed either in endothelial cells (Tie2-Cre) or in smooth muscle cells and pericytes (␣-smooth muscle actin [␣sma] Cre) to mouse models of pathological neovascularization.ADAM17 was first discovered as the converting enzyme for tumor necrosis factor (TNF)␣, 8,9 a potent proinflammatory cytokine that is a causative factor in autoimmune diseases such as rheumatoid arthritis and Crohn's disease as well as in septic shock in mice. 10 Once mice lacking ADAM17 were generated, it became clear that ADAM17 is Original received August 14, 2009; revision received January 13, 2010; accepted January 19, 2010 The ability of ADAM17 to release endothelial cell membrane proteins on stimulation with VEGF-A raised questions about what role ADAM17 has during developmental angiogenesis and in pathological neovascularization in adult animals. Although mice lacking ADAM17 die perinatally, most likely as a consequence of their severe heart valve defects, 11,12 there have been no reports of defects in developmental angiogenesis in these animals. To address whether ADAM17 has a role in angiogenesis or pathological neovascularization or both, we conditionally inactivated ADAM17 in endothelial cells or in smooth muscle cells such as pericytes, and then determined how lack of ADAM17 affects two mouse models for pathological neovascularization, the oxygen induced retinopathy model for retinopathy of prematurity, and growth of heterotopically injected tumor cells. Moreover, we assessed proliferation and chord formation of endothelial cells lacking ADAM17, and evaluated the role of ADAM17 in the proteolytic release of membrane proteins with known roles in angiogenesis and pathological neovascularization. Methods Reagents and Cell LinesPorcine aortic endothelial cells expressing VEGFR2/KDR (PAE/ KDR cells) and mouse embryonic fibroblasts (mEFs) lacking ADAM17 have been described previously. 6,15 Reagents were from...

The angiotensin II (AngII) type 1 receptor (AT(1)) plays a critical role in hypertrophy of vascular smooth muscle cells (VSMCs). Although it is well known that G(q) is the major G protein activated by the AT(1) receptor, the requirement of G(q) for AngII-induced VSMC hypertrophy remains unclear. By using cultured VSMCs, this study examined the requirement of G(q) for the epidermal growth factor receptor (EGFR) pathway, the Rho-kinase (ROCK) pathway, and subsequent hypertrophy. AngII-induced intracellular Ca(2+) elevation was completely inhibited by a pharmacological G(q) inhibitor as well as by adenovirus encoding a G(q) inhibitory minigene. AngII (100nm)-induced EGFR transactivation was almost completely inhibited by these inhibitors, whereas these inhibitors only partially inhibited AngII (100nm)-induced phosphorylation of a ROCK substrate, myosin phosphatase target subunit-1. Stimulation of VSMCs with AngII resulted in an increase of cellular protein and cell volume but not in cell number. The G(q) inhibitors completely blocked these hypertrophic responses, whereas a G protein-independent AT(1) agonist did not stimulate these hypertrophic responses. In conclusion, G(q) appears to play a major role in the EGFR pathway, leading to vascular hypertrophy induced by AngII. Vascular G(q) seems to be a critical target of intervention against cardiovascular diseases associated with the enhanced renin-angiotensin system.

A metalloprotease, ADAM17, mediates the generation of mature ligands for the epidermal growth factor receptor (EGFR). This is the key signaling step by which angiotensin II (AngII) induces EGFR transactivation leading to hypertrophy and migration of vascular smooth muscle cells (VSMCs). However, the regulatory mechanism of ADAM17 activity remains largely unclear.Here we hypothesized that caveolin-1 (Cav1), the major structural protein of a caveolae, a membrane microdomain, is involved in the regulation of ADAM17. In cultured VSMCs, infection of adenovirus encoding Cav1 markedly inhibited AngII-induced EGFR ligand shedding, EGFR transactivation, ERK activation, hypertrophy and migration, but not intracellular Ca 2+ elevation. Methyl-β-cyclodextrin and filipin, reagents that disrupt raft structure, both stimulated an EGFR ligand shedding and EGFR transactivation in VSMCs. In addition, non-detergent sucrose gradient membrane fractionations revealed that ADAM17 cofractionated with Cav1 in lipid rafts. These results suggest that lipid rafts and perhaps caveolae provide a negative regulatory environment for EGFR transactivation linked to vascular remodeling induced by AngII. These novel findings may provide important information to target cardiovascular diseases under the enhanced renin angiotensin system.

To develop a wearable multi-lead electrocardiogram (ECG) measuring system, we fabricated the electrodes and wires by using electrostatic flocking technology on a textile. By using this technology, it was possible to fabricate many electrodes and wires, simultaneously. Also the flocked electrodes and wires had stretchability and washing resistance properties. To use dry electrodes, it is important to reduce the influence of motion artifacts (MAs). The results of the experiment with the skin phantom revealed that the contact pressure between the skin and the electrode is an important factor in MA reduction. Then, we conducted experiments with a human body to determine the relationship between the contact pressure and the MAs. Under the pressures of 200 Pa and 500 Pa, MAs were observed. Meanwhile, under the pressures of 1000 Pa, 2000Pa and 4000 Pa, the ECG signals under rest and deep breathing conditions were able to be measured without MAs. Considering the comfortability, the contact pressure from 1000 Pa to 2000 is preferable. Finally, we fabricated the wearable ECG measuring system and succeeded in measuring 18-lead ECG signals. The measured ECG waveform is in good agreement with the ECG waveform measured by a commercial system.

Through our study, greater improvement of postoperative fecal continence after LAARP has not been shown. LAARP was at higher risk for mucosal prolapse and PUD. However, precise dissection of the urethral fistula could be performed after the introduction of urethroscopy.

Background-Although, endothelial nitric oxide (NO) synthase (eNOS) is believed to antagonize vascular remodeling induced by the angiotensin II (AngII) type-1 receptor, the exact signaling mechanism remains unclear. Methods and Results-By expressing eNOS to vascular smooth muscle cells (VSMCs) via adenovirus, we investigated a signal transduction mechanism of the eNOS gene transfer in preventing vascular remodeling induced by AngII. We found marked inhibition of AngII-induced Rho/Rho-kinase activation and subsequent VSMC migration by eNOS gene transfer whereas G q -dependent transactivation of the epidermal growth factor receptor by AngII remains intact. This could be explained by the specific inhibition of G 12/13 activation by eNOS-mediated G 12/13 phosphorylation. Conclusion-The eNOS/NO cascade specifically targets the Rho/Rho-kinase system via inhibition of G 12/13 to prevent vascular migration induced by AngII, representing a novel signal cross-talk in cardiovascular protection by NO.

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Summary The requirement of a metalloprotease ADAM17 (a disintegrin and metalloprotease 17) for the growth of cultured vascular smooth muscle cells has been demonstrated in vitro. However, whether this metalloprotease is responsible for vascular remodeling in vivo remains unanswered. Rat carotid arteries were analyzed 2 weeks after a balloon angioplasty. The neointimal cells were strongly positive for ADAM17 immunostaining. Marked inhibition of intimal hyperplasia was observed in dominant negative ADAM17 adenovirus treated carotid artery. Proliferating cell nuclear antigen positive cells and phospho-epidermal growth factor receptor positive cells in the neointima were reduced by dominant negative ADAM17 as well. In contrast, the neointima formation, proliferating cell nuclear antigen positive cells, and phospho-epidermal growth factor receptor positive cells were markedly enhanced by wild-type ADAM17 adenovirus. In conclusion, ADAM17 activation is involved in epidermal growth factor receptor activation and subsequent neointimal hyperplasia after vascular injury. ADAM17 could be a novel therapeutic target for pathophysiological vascular remodeling.