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Cyclosporin (Sandimmun?) plasma concentrations were measured in the Department of Hematology and Oncology��s routine laboratory using a fluoroscopic polarization immunoassay (TDx/TDxFLx? Cyclosporin Monoclonal Whole Blood Assay; Abbott Laboratories, Abbott Park, IL, USA). Everolimus (Certican?) plasma concentrations were also measured using a fluoroscopic polarization immunoassay (INNOFLUOR?-CERTICAN?-Assay; https://www.selleckchem.com/products/obeticholic-acid.html Seradyn, Indianapolis, IN, USA). For both analyses, each 150?��l of mouse blood was harvested from the plexus retroorbitalis mixed with 50?��l ethylene diamine acetic acid (EDTA) and immediately transferred for analysis. Every sample was analysed in triplicates. The procedure was performed using a modified technique initially described by Koulack et?al. [16]. In brief, the donor thoracic aorta was isolated, resected and transferred to the recipient animal. The recipient aorta was clamped and then transected with sharp microvascular scissors. A proximal end to end anastomosis was performed. The aortic graft was then repositioned and the anastomosis continued with single interrupted sutures. Aortic grafts were removed under anaesthesia on day 30 after transplantation. Grafts were perfused with saline and were flash frozen in optimal cutting temperature (OCT) medium (Tissue-Tek?; Sakura, The Netherlands) in liquid nitrogen for morphometric analysis of 7?��m cryostat sections. A minimum of five transverse sections was analysed from each graft. Five sections from each graft harvested at day 30 were stained with Elastin/van Gieson and analysed at an original magnification of ��200 using https://www.selleck.cn/products/LY294002.html a conventional light microscope. A digitized image of each section was captured and the areas within the lumen and the internal elastic lamina were circumscribed manually and measured as previously described [17]. All image analyses were carried out on a colour display monitor using ANAlysis? Image Analysis software (Olympus, Hamburg, Germany). Results are given as the mean per group?��?SD which was derived from the mean per graft. Statistical analysis was performed using a one-way https://www.selleckchem.com/products/Adriamycin.html anova followed by a Bonferroni correction. Differences between groups are considered as significant when P?