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Title:           Targeting LDL aggregation decreases atherosclerotic lipid burden in a humanized mouse model of familial hypercholesterolemia: Crucial role of ApoB100 conformational stabilization
Subject:         Atherosclerosis, 410 (2025) 118630. doi:10.1016/j.atherosclerosis.2024.118630
Keywords:        Atherosclerosis,LDL aggregation,LRP1-Based peptides,ApoB100 conformation,Therapeutic peptides
Author:          A. Benitez-Amaro
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      Title: Targeting LDL aggregation decreases atherosclerotic lipid burden in a humanized mouse model of familial hypercholesterolemi ...
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        Title: 1 Introduction
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        Title: 2 Materials and methods
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          Title: 2.1 Animal studies
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            Title: 2.1.1 Generation of the animal model
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            Title: 2.1.2 Experimental design for induction and treatment of atherosclerosis in vivo
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            Title: 2.1.3 In vivo biodistribution of DP3 peptide
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            Title: 2.1.4 Anesthesia and euthanasia
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          Title: 2.2 Characteristics of patients with hypercholesterolemia
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          Title: 2.3 Peptide synthesis, labeling, and purification
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          Title: 2.4 Blood collection from FH patients
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          Title: 2.5 Blood collection and monocyte quantification in the in vivo model
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          Title: 2.6 Culture of smooth muscle cells (SMC)
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          Title: 2.7 Biochemical studies
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          Title: 2.8 Polyacrylamide gradient gel electrophoresis
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          Title: 2.9 Reverse transcription and real-time qPCR analysis
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          Title: 2.10 Thin layer chromatography of neutral lipid content in smooth muscle cells (SMCs) and murine livers
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          Title: 2.11 Bioimaging evaluation of atherosclerotic lipid burden
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          Title: 2.12 Studies about the susceptibility of LDL to aggregation
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            Title: 2.12.1 Lipoprotein isolation and purification
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            Title: 2.12.2 Study of passive and sphingomyelinase (SMase)-induced LDL aggregation
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            Title: 2.12.3 Particle size distribution (PSD) measuraments in human LDL samples from patients with familial hypercholesterolemia
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            Title: 2.12.4 Study of proteoglycan (PG)-induced LDL aggregation
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            Title: 2.12.5 High-performance thin layer chromatography (HPTLC) of phospholipid lipid content in murine and human LDL samples
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            Title: 2.12.6 Agarose gel electrophoresis of murine and human LDL
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            Title: 2.12.7 Z-potential measurements to identify changes in LDL particle charge induced by DP3 binding
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            Title: 2.12.8 Transmission electron microscopy (TEM) of murine LDL samples
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            Title: 2.12.9 Circular Dichroism of murine LDL to identify conformational ApoB100 changes induced by peptides
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            Title: 2.12.10 SDS-PAGE and Western blot analysis of murine LDL
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          Title: 2.13 Statistical analysis
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        Title: 3 Results
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          Title: 3.1 Characterization of lipid and lipoprotein profile of Ldlr−/− hApoB100 mice
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          Title: 3.2 Imaging monitoring of TAMRA-DP3 biodistribution in Ldlr−/− hApoB100 mice
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          Title: 3.3 Effects of DP3 in HFD-induced vascular lipid accumulation in Ldlr−/− hApoB100 mice
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          Title: 3.4 Effect of peptide administration in systemic and hepatic inflammation
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          Title: 3.5 Susceptibility of LDL isolated from DP3-treated (compared to that from control groups) to passive (no SMase) and SMase- ...
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          Title: 3.6 Effect of DP3 on ApoB100 structural conformation in LDL
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          Title: 3.7 Effects of DP3 ex-vivo on human LDL tendency to aggregation. Studies in LDL isolated from patients with familiar hyperc ...
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        Title: 4 Discussion
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        Title: Author contributions
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        Title: Data availability
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        Title: Financial support
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        Title: Declaration of competing interest
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        Title: Acknowledgements
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        Title: Appendix A Supplementary data
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        Title: References
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