6 research outputs found

    Characterization of Carotid Smooth Muscle Cells during Phenotypic Transition

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    Vascular smooth muscle cells (VSMCs) are central players in carotid atherosclerosis plaque development. Although the precise mechanisms involved in plaque destabilization are not completely understood, it is known that VSMC proliferation and migration participate in plaque stabilization. In this study, we analyzed expression patterns of genes involved in carotid atherosclerosis development (e.g., transcription factors of regulation of SMC genes) of VSMCs located inside or outside the plaque lesion that may give clues about changes in phenotypic plasticity during atherosclerosis. VSMCs were isolated from 39 carotid plaques extracted from symptomatic and asymptomatic patients by endarterectomy. Specific biomarker expression, related with VSMC phenotype, was analyzed by qPCR, western immunoblot, and confocal microscopy. MYH11, CNN1, SRF, MKL2, and CALD1 were significantly underexpressed in VSMCs from plaques compared with VSMCs from a macroscopically intact (MIT) region, while SPP1, KLF4, MAPLC3B, CD68, and LGALS3 were found significantly upregulated in plaque VSMCs versus MIT VSMCs. The gene expression pattern of arterial VSMCs from a healthy donor treated with 7-ketocholesterol showed high similarity with the expression pattern of carotid plaque VSMCs. Our results indicate that VSMCs isolated from plaque show a typical SMC dedifferentiated phenotype with macrophage-like features compared with VSMCs isolated from a MIT region of the carotid artery. Additionally, MYH11, KLF5, and SPP1 expression patterns were found to be associated with symptomatology of human carotid atherosclerosis.This work was supported by Spanish Institute for Health Carlos III, RETICS program (Grant Number RD16/0019/0007) and by Basque Government, Education Department, Consolidated Groups program (Grant Number IT512-10)

    Pathophysiology of Atherosclerosis

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    Atherosclerosis is the main risk factor for cardiovascular disease (CVD), which is the leading cause of mortality worldwide. Atherosclerosis is initiated by endothelium activation and, followed by a cascade of events (accumulation of lipids, fibrous elements, and calcification), triggers the vessel narrowing and activation of inflammatory pathways. The resultant atheroma plaque, along with these processes, results in cardiovascular complications. This review focuses on the different stages of atherosclerosis development, ranging from endothelial dysfunction to plaque rupture. In addition, the post-transcriptional regulation and modulation of atheroma plaque by microRNAs and lncRNAs, the role of microbiota, and the importance of sex as a crucial risk factor in atherosclerosis are covered here in order to provide a global view of the disease.This work was supported by the Basque Government (Grupos Consolidados IT-1264-19). A.B.-V. was supported by Programa de especializaci├│n de Personal Investigador Doctor en la UPV/EHU (2019) 2019/2020; U.G-G. was supported by Margarita Salas Grant; and S.J. and A.L-S were supported by a grant PIF (2017ÔÇô2018) and PIF (2019ÔÇô2020) Gobierno Vasco, respectively

    Cholesterol Efflux Efficiency of Reconstituted HDL Is Affected by Nanoparticle Lipid Composition

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    Cardiovascular disease (CVD), the leading cause of mortality worldwide is primarily caused by atherosclerosis, which is promoted by the accumulation of low-density lipoproteins into the intima of large arteries. Multiple nanoparticles mimicking natural HDL (rHDL) have been designed to remove cholesterol excess in CVD therapy. The goal of this investigation was to assess the cholesterol efflux efficiency of rHDLs with different lipid compositions, mimicking different maturation stages of high-density lipoproteins (HDLs) occurring in vivo. Methods: the cholesterol efflux activity of soybean PC (Soy-PC), 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC), DPPC:Chol:1-palmitoyl-2-hydroxy-sn-glycero-3-phosphocholine (LysoPC) and DPPC:18:2 cholesteryl ester (CE):LysoPC rHDLs was determined in several cell models to investigate the contribution of lipid composition to the effectiveness of cholesterol removal. Results: DPPC rHDLs are the most efficient particles, inducing cholesterol efflux in all cellular models and in all conditions the effect was potentiated when the ABCA1 transporter was upregulated. Conclusions: DPPC rHDLs, which resemble nascent HDL, are the most effective particles in inducing cholesterol efflux due to the higher physical binding affinity of cholesterol to the saturated long-chain-length phospholipids and the favored cholesterol transfer from a highly positively curved bilayer, to an accepting planar bilayer such as DPPC rHDLs. The physicochemical characteristics of rHDLs should be taken into consideration to design more efficient nanoparticles to promote cholesterol efflux.This work was supported by the Basque Government (Grupos Consolidados IT-1264-19). U.G.-G. was supported by Fundaci├│n Biof├şsica Bizkaia. A.B.-V. was supported by Programa de especializaci├│n de Personal Investigador Doctor en la UPV/EHU (2019) 2019ÔÇô2020. S.J.-B. and A.L.-S. were supported by a grant PIF (2017ÔÇô2018) and (2019ÔÇô2020), Gobierno Vasco, respectively. A.L.-S. was partially supported by Fundaci├│n Biof├şsica Bizkaia

    BIRC6 Is Associated with Vulnerability of Carotid Atherosclerotic Plaque

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    Carotid atherosclerotic plaque rupture can lead to cerebrovascular accident (CVA). By comparing RNA-Seq data from vascular smooth muscle cells (VSMC) extracted from carotid atheroma surgically excised from a group of asymptomatic and symptomatic subjects, we identified more than 700 genomic variants associated with symptomatology (p < 0.05). From these, twelve single nucleotide polymorphisms (SNPs) were selected for further validation. Comparing genotypes of a hospital-based cohort of asymptomatic with symptomatic patients, an exonic SNP in the BIRC6 (BRUCE/Apollon) gene, rs35286811, emerged as significantly associated with CVA symptomatology (p = 0.002; OR = 2.24). Moreover, BIRC6 mRNA levels were significantly higher in symptomatic than asymptomatic subjects upon measurement by qPCR in excised carotid atherosclerotic tissue (p < 0.0001), and significantly higher in carriers of the rs35286811 risk allele (p < 0.0001). rs35286811 is a proxy of a GWAS SNP reported to be associated with red cell distribution width (RDW); RDW was increased in symptomatic patients (p < 0.03), but was not influenced by the rs35286811 genotype in our cohort. BIRC6 is a negative regulator of both apoptosis and autophagy. This work introduces BIRC6 as a novel genetic risk factor for stroke, and identifies autophagy as a genetically regulated mechanism of carotid plaque vulnerability.This work was financially supported by grants from the Departments of Education (Ref. PIBA2018-67) and Health (Ref. RIS3-2019222038) of the Basque Government, Vitoria-Gasteiz, Spain; by the Spanish Neurovascular Network (INVICTUSplus) (Ref. RD16/0019/0007) funded by the Instituto de Salud Carlos III, Ministerio de Ciencia e Innovaci├│n, Madrid, Spain; and by the research project grant (IKERIKTUS) funded by the RefbioII Trans-Pyrenean Cooperation Network for Biomedical Research financed by Horizon 2020. I.A. is supported by the Marat├│n EiTB 2017 for Funding of Research into Stroke, Bilbao, Spain (Ref. BIO18/IC/005); R.T.N. is the recipient of a fellowship from the Secretar├şa Nacional de Ciencia y Tecnolog├şa e Innovaci├│n (SENACYT; Convocatoria Doctorado de Investigaci├│n Ronda III, 2018; Ref. BIDP-III-2018-12) of the Gobierno Nacional, Rep├║blica de Panam├í

    Interactome of the Autoimmune Risk Protein ANKRD55

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    The ankyrin repeat domain-55 (ANKRD55) gene contains intronic single nucleotide polymorphisms (SNPs) associated with risk to contract multiple sclerosis, rheumatoid arthritis or other autoimmune disorders. Risk alleles of these SNPs are associated with higher levels of ANKRD55 in CD4(+) T cells. The biological function of ANKRD55 is unknown, but given that ankyrin repeat domains constitute one of the most common protein-protein interaction platforms in nature, it is likely to function in complex with other proteins. Thus, identification of its protein interactomes may provide clues. We identified ANKRD55 interactomes via recombinant overexpression in HEK293 or HeLa cells and mass spectrometry. One hundred forty-eight specifically interacting proteins were found in total protein extracts and 22 in extracts of sucrose gradient-purified nuclei. Bioinformatic analysis suggested that the ANKRD55-protein partners from total protein extracts were related to nucleotide and ATP binding, enriched in nuclear transport terms and associated with cell cycle and RNA, lipid and amino acid metabolism. The enrichment analysis of the ANKRD55-protein partners from nuclear extracts is related to sumoylation, RNA binding, processes associated with cell cycle, RNA transport, nucleotide and ATP binding. The interaction between overexpressed ANKRD55 isoform 001 and endogenous RPS3, the cohesins SMC1A and SMC3, CLTC, PRKDC, VIM, beta-tubulin isoforms, and 14-3-3 isoforms were validated by western blot, reverse immunoprecipitaton and/or confocal microscopy. We also identified three phosphorylation sites in ANKRD55, with S436 exhibiting the highest score as likely 14-3-3 binding phosphosite. Our study suggests that ANKRD55 may exert function(s) in the formation or architecture of multiple protein complexes, and is regulated by (de)phosphorylation reactions. Based on interactome and subcellular localization analysis, ANKRD55 is likely transported into the nucleus by the classical nuclear import pathway and is involved in mitosis, probably via effects associated with mitotic spindle dynamics.This work was supported by the following grants to KV: Grupos de Investigacion (IT512-10, PPG17/44) and MINECO (SAF2016-74891R). NU is recipient of a predoctoral studentship from the Gobierno Vasco (Reference PRE-2013-1-891). CIC bioGUNE is accredited with the Severo Ochoa Excellence award by the Spanish Ministerio de Econom i a y Competitividad, MINECO (SEV-2016-0644)

    Evaluation Of The Cases With Intracranial Hypertension

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    Ama├ž: D├╝zce ├ťniversitesi Ara┼čt─▒rma ve Uygulama Hastanesi, N├Âroloji klini─činde ─░HH tan─▒s─▒ konulmu┼č hastalar─▒n klinik bulgu ve tedavilerinin prognoz ile ili┼čkilerinin incelenmesi ama├žlanm─▒┼čt─▒r. Gere├ž ve Y├Ântem: ─░─░H tan─▒s─▒ ile tedavi ve takip edilen 23 hasta incelendi. ├çal─▒┼čmada Modifiye Dandy Kriterleri esas al─▒nd─▒. Kranial g├Âr├╝nt├╝leme yap─▒larak, lomber ponksiyonlar─▒ ger├žekle┼čtirildi. Bulgular: ─░─░H tan─▒l─▒ hastalar─▒n %78,3'si kad─▒n, %21,7 si erkekti. En s─▒k ba┼čvuru nedeni olan ba┼č a─čr─▒s─▒na, bulan─▒k g├Ârme, bak─▒┼č k─▒s─▒tl─▒l─▒─č─▒, ge├žici g├Ârme kayb─▒, g├Âz a─čr─▒s─▒, ├žift g├Ârme, bulant─▒, ─▒┼č─▒ktan rahats─▒z olma, ba┼č d├Ânmesi ve kulak ├ž─▒nlamas─▒ e┼člik ediyordu. Hastalar─▒n %60.9'u obezdi. Be┼č hastada papil ├Âdem g├Âzlenmeksizin ─░─░H tan─▒s─▒ saptand─▒.Empty sella d─▒┼č─▒nda kranial g├Âr├╝nt├╝leme normal s─▒n─▒rlardayd─▒. Tedavide, asetozolamid, metilprednisolon ve/veya topiramat verildi. Takip s├╝resi 3-6 ay olarak d├╝zenlendi. Bu s├╝re├ž i├žinde g├Ârme kayb─▒ ya┼čayan hastam─▒z olmad─▒. Sonu├ž: Devaml─▒l─▒k g├Âsteren atipik ba┼č a─čr─▒ vakalar─▒nda, obezite varsa ─░─░H tan─▒s─▒ d├╝┼č├╝n├╝lerek ileri tetkik yap─▒lmal─▒d─▒r. Erken tedavi, olas─▒ g├Ârme kay─▒plar─▒n─▒ ├Ânlemede ├Ânem ta┼č─▒maktad─▒r.Objective: In this study clinical findings of patients, who diagnosed with IIH in Duzce Univesity Research and Teaching Hospital Neurology Clinic were investigated. Materials and Methods: Treatment and follow-up of 23 patients were examined with diagnosis of IIH from the records. The study was based on modified Dandy criteria. The patients who were performed lumbar puncture and were done cranial imaging included in the study. Results: The patients diagnosed with IIH were 78.3% female and 21.7% male. The most common reason for admission was headache and it was accompanied by blurred vision, visual of limitation, temporary loss of vision, eye pain, double vision, nausea, dislike of light, dizziness and tinnitus. 60.9% of the patients were obese. Five patients without papilledema were diagnosed with IIH. The cranial imagings were in normal limits except for empty cella. In treatment, the patients were given acetozolamide, methylprednisolone and/or topiramate. The follow-up period was arranged in 3-6 months. In the process, there were not any patients who had loss of vision. Conclusion: Continuity in atypical cases of headache, if they have obesity, there should be further examination in mind of an IIH diagnosis. Early diagnosis and treatment are import to prevent the possible loss of vision