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1. IASG Romanian Society of Surgery Romtransplant The V-th Symposium and P ost graduate Course of IASG Bucharest 9-11 April 2003
2. “ DOMINO” LIVER TRANSPLANTATION WITH PARTICULAR INDICATION (FAMILIAL HYPERCHOLESTEROLEMIA) I POPESCU, M SIMIONESCU, D TULBURE, A SIMA, C CATANA, L NICULESCU, N HANCU, L GHEORGHE, M MIHAILA, S CIUREA, V VIDU, D HREHORET CENTER OF GENERAL SURGERY AND LIVER TRANSPLANTATION DEPARTMENT OF INTENSIVE CARE AND ANESTHESIOLOGY CENTER OF GASTROENTEROLOGY FU N DENI CLINICAL INSTITUTE INSTITUTE OF CELLULAR BIOLOGY AND PATHOLOGY "NICOLAE SIMIONESCU“ INSTITUTE OF DIABETES AND NUTRITIONAL DISEASES CLUJ-NAPOCA
37. Quantification of the expression of LDL-R on monocytes by flow-cytometry analysis, one year after the transplant operations *mean value given by LDL receptor kit 71 153.5 Patient B (Cirrhosis) 6.7 14.4 Patient A (FHC) 100.0 130 - 277 (mean 216) Control* LDL-R (%) Mean fluorescence (arbitrary units) Monocytes
38. Histogram showing quantification of LDL receptors on monocytes isolated from patient A (familial hypercholesterolemia) and B (cirrhosis) by flow cytometry, using an anti-LDL receptor monoclonal antibody
39. Gene expression of LDL-R of monocyte-derived macrophages isolated from patient A (familial hypercholesterolemia) and B (cirrhosis); agarose gel electrophoresis of RT-PCR products. Note the presence of LDL receptor (LDL-R) expression in both cases, although apparently reduced in the case of patient B due to the reduced activation of monocytes. GAPDH and LDL-R gene expression are identified by comparing with a ladder DNA (lane 1)
40. Gene expression of LDL-R in fibroblasts obtained from patient A (familial hypercholesterolemia) and B (cirrhosis). Agarose gel electrophoresis of RT-PCR products of RNA isolated from primary culture fibroblasts from the two subjects. Expression of mRNA coding for the LDL receptor (LDL-R) as compared with the GAPDH expression. Lane 1: ladder DNA
A girl aged 6 years 9 months with severe heart disease secondary to homozygous familial hypercholesterolaemia underwent orthotopic cardiac transplantation and her liver was replaced with the liver of the same donor. In the first 10 weeks after transplantation serum cholesterol fell to 270 mg/dl from preoperative concentrations of more than 1000 mg/dl
Flow-cytometry analysis allowed the quantification of LDL receptors on monocytes isolated from both patients as compared with the mean value given by the kit
The amplified sequences coding for LDL-R and GAPDH from each patient were visualized following their electrophoretic migration in 1% agarose gel. As shown in Figure 4, the MDM obtained from FHC patient presented normal LDL-R gene expression. The domino recipient patient expressed LDL-R as well, but the expression was apparently diminished due to the fact that, in this case, the activation of monocytes was slightly reduced and this may explain the apparently diminished LDL-R expression in MDM obtained from the recipient patient (Fig.4).
These results were confirmed by the RT-PCR products from RNA isolated from primary culture fibroblasts obtained from the skin explants of patients A and B. The data showed the positive expression of mRNA coding for LDL-R in the fibroblasts of the two patients (Fig. 5).
