Citation for published version (APA): Sivapalaratnam, S. (2012). The molecular basis of early onset cardiovascular disease

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1 UvA-DARE (Digital Academic Repository) The molecular basis of early onset cardiovascular disease Sivapalaratnam, S. Link to publication Citation for published version (APA): Sivapalaratnam, S. (2012). The molecular basis of early onset cardiovascular disease General rights It is not permitted to download or to forward/distribute the text or part of it without the consent of the author(s) and/or copyright holder(s), other than for strictly personal, individual use, unless the work is under an open content license (like Creative Commons). Disclaimer/Complaints regulations If you believe that digital publication of certain material infringes any of your rights or (privacy) interests, please let the Library know, stating your reasons. In case of a legitimate complaint, the Library will make the material inaccessible and/or remove it from the website. Please Ask the Library: or a letter to: Library of the University of Amsterdam, Secretariat, Singel 425, 1012 WP Amsterdam, The Netherlands. You will be contacted as soon as possible. UvA-DARE is a service provided by the library of the University of Amsterdam ( Download date: 25 Jul 2018

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4 269 Scientist Come out of Your Lab! Jurriaan van Hall, April 2010, Amsterdam Painted at De Hoop during the first APROVE PhD Symposium Oil on Canvas, 150 x 250 cm Amsterdam, AMC, E2 PROLOGUE R1 R2 R3 R4 R5 R6 R7 R8 R9 R10 R11 R12 R13 R14 R15 R16 R17 R18 R19 R21 R22 R23 R24 R25 R26 R27 R28 R29 R30 R31 R32 R33 R34 R35 R36 R37 R38 R39

5 CHAPTER 4 Figure 2. Reclassification based on family history of premature coronary heart disease 270

6 CHAPTER 6 Figure 1 Liver Atherosclerotic plaque ANRIL PCSK9 SORT1 MRAS LPA LDL-R SH2B3 S LDL PHACTR1 P CLCX12 apob100 ob100 VLDL apoe ap apoc C Apo denotes apolipoprotein; LDL denotes low-density lipoprotein; VLDL denotes very low-density lipoprotein; LDL-R denotes low-density lipoprotein-receptor. R1 R2 R3 R4 R5 R6 R7 R8 R9 R10 R11 R12 R13 R14 R15 R16 R17 R18 R19 CHAPTER 9 Figure 2. Manhattan plots for HDL-C. LDL-C, TC and TG data from the International IBC lipid metaanalysis based on estimates obtained using a p-value based meta-analysis in METAL 271 R21 R22 R23 R24 R25 R26 R27 R28 R29 R30 R31 R32 R33 R34 R35 R36 R37 R38 R39

7 CHAPTER 10 Supplementary Figure 1. Results of parametric linkage analysis A significant LOD score of 3.31 was obtained on chromosome 12 after parametric linkage analysis by Allegro software using genotypes from 12 available family members. Figure 2. Keratocan is expressed in atherosclerotic but not normal human coronary arteries Macrophages are stained red and Keratocan is stained blue. 2a shows a normal coronary artery; macrophages and Keratocan are absent. 2b shows a fatty streak; macrophages entered into the vessel wall and Keratocan is present. 2c shows an atherosclerotic plaque; Keratocan is strongly expressed in the lipid core. 272

8 Supplementary Figure 2. Triple staining for Keratocan, CXCL-1 and CD3+. Triple staining for CXCL-1 (red); KERA (blue) and CD3+ cells (brown) demonstrate that these cells are positive for all three components. As described in the literature CXCL-1 is clearly present in the atherosclerotic vessel wall. Supplementary Figure 3. Western blot of Keratocan in human plaques and cornea Keratocan was seen as a ~53kDa band after western blotting using Keratocan antibody. Cornea tissue was used as positive control. 273 R1 R2 R3 R4 R5 R6 R7 R8 R9 R10 R11 R12 R13 R14 R15 R16 R17 R18 R19 R21 R22 R23 R24 R25 R26 R27 R28 R29 R30 R31 R32 R33 R34 R35 R36 R37 R38 R39

9 Supplementary Figure 4. Keratocan is present in atherosclerotic mice Early and advanced atherosclerotic tissue from murine carotid arteries were stained using Keratocan antibody (brown) and Hematoxylin as a counterstaining (blue). While present mainly near endothelial cells in early lesion, Keratocan is predominantly present in the matrix of the plaque in advanced lesions, suggesting production by endothelial cells and secretion in the matrix. 274

10 CHAPTER 11 Figure 1. Pedigree of the family with early onset CAD Arrow for the index case.the right half of a symbol denotes CVD status. If fully colored this indicates CVD. On the left half, the presence of mutation in ZC3HC1 and MCF2L is indicated with green (top corner) and yellow (bottom corner) colors respectively. The first number is an identifier within the generation. The number between brackets is the current age. AMI denotes Acute Myocardial Infarction; AP denotes Angina Pectoris, PTCA denotes Percutaneous Transluminal Coronary Angiogrpahy; CABG denotes Coronary Artery Bypass Graft; this line also contains the age of event in years. RF denotes Risk Factors at time of event; py denotes cigarette pack years; HT denotes Hypertension, None of the subjects suffered from diabetes mellitus or was obese. FH was excluded in the index case (arrow). Subsequently, the lipid profiles are displayed; TC, HDL-c, LDL-c and TG in mmol/l. On the final line Ator denotes atorvastatin; Simva denotes simvastatin and Ez denotes ezetimibe treatment. 275

11 R1 R2 R3 R4 R5 R6 R7 R8 R9 R10 R11 R12 R13 R14 R15 R16 R17 R18 R19 R20 R21 R22 R23 R24 R25 R26 R27 R28 R29 R30 R31 R32 R33 R34 R35 R36 R37 R38 R39 276

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