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A. Rabbit Polyclonal to INTS2 development of vascular remodeling. deficient (wild-type (test using the SPSS 17.0 software. mice at the age of 12 weeks. The left external jugular vein from a C57 mouse was dissected from the main trunk. A segment of the jugular vein (5 mm long) was sutured into place to repair a longitudinal defect (about the same length as the jugular vein patch) in the left carotid artery from mouse. The grafts, together with a short segment of the native carotid artery, were harvested at 4 weeks after surgery by cutting at the center of the graft. One portion was processed for paraffin embedding and the other for cryostat sectioning. A. Schema of the vein graft and cross-section of the composite vessel at the center of the graft. B. Verhoeff-van Gieson (VVG) staining in paraffin-embedded cross-section from center of the graft of mice at 4 weeks post-operation. 3C4 JNJ-42165279 layers of elastic lamina mark the artery. The vein is greatly enlarged in response to arterial blood pressure. C. -smooth muscle actin (-SMA) immunostaining in vein graft cryostat section from mice at 4 weeks post-operation. Vascular smooth muscle cell appears red. D. Verhoeff-van Gieson (VVG) staining in paraffin embedded cross-section of aorta from normal mice. Notice that normal artery has well-organized and circumferentially-oriented elastin. Open in a separate window Number 2 Severe HHcy promotes vascular redesigning and elastin production in the neointima in the vein graft in miceThe mouse vein graft surgery process was performed on mice at the age of 12 weeks as explained in Number 1 and in the section of Methods. The vein patch grafts, together with a short section of the native carotid artery, were harvested at 4 weeks after surgery and processed JNJ-42165279 for paraffin embedding (VVG) and cryostat sections (Cactin). Mouse plasma was collected at the end of each experiment (at age 16 weeks). A. Photomicrographs of morphological analysis. Sections are oriented having a vein patch on top and artery on bottom. VVG staining shows elastin as black. Anti–mouse clean muscle mass (-SMA) antibody staining shows clean muscle mass cell as reddish. B. Quantitative analysis. Neointimal area and thickness were analyzed by computerized planimetry using the updated Image-Pro Plus system. We identified the elastin positive area by computer-assisted color gated measurement (Image-Pro Plus) and indicated the data as a percentage. The neointima was defined as the region between the internal elastic lamina (IEL) and the lumen. The percentage luminal narrowing was determined as 100 x (the difference between area inside the IEL and area of the lumen the area inside the IEL. Hcy significantly improved neointima formation, luminal narrowing and neointima elastin JNJ-42165279 deposition. C. Plasma level of Hcy. Hcy concentration was measured by liquid chromatography electrospray tandem mass spectrometry methods. Values represent meanSEM, n=9, ideals from self-employed t test * p 0.05 versus group, #p 0.001 versus group. VVG, Verhoeff-van Gieson staining. 4.2 HHcy increases neointimal elastin deposition in miceThe mouse vein graft surgery process was performed on Cbs mice at the age of 12 weeks as explained in Number 1 and in the section of Methods. The grafts, together with a short section of the native carotid artery, were harvested at 4 weeks after surgery by trimming at the center of the graft. One portion was processed for paraffin embedding and the additional for cryostat.