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1.
J Immunol ; 198(8): 3109-3117, 2017 04 15.
Artículo en Inglés | MEDLINE | ID: mdl-28264972

RESUMEN

The development of rheumatoid arthritis (RA) is linked to functional changes in synovial fibroblasts (SF) and local infiltration of T lymphocytes. Fibroblasts possess the capacity to suppress T cell responses, although the molecular mechanisms of this suppression remain incompletely understood. In this study, we aimed to define the mechanisms by which noninflammatory SF modulate Th cell responses and to determine the immunosuppressive efficacy of RASF. Hence, the influence of SF from osteoarthritis or RA patients on total Th cells or different Th cell subsets of healthy donors was analyzed in vitro. We show that SF strongly suppressed the proliferation of Th cells and the secretion of IFN-γ in a cell contact-independent manner. In cocultures of SF and Th cells, tryptophan was completely depleted within a few days, resulting in eukaryotic initiation factor 2α phosphorylation, TCRζ-chain downregulation, and proliferation arrest. Blocking IDO1 activity completely restored Th cell proliferation, but not IFN-γ production. Interestingly, only the proliferation of Th1 cells, but not of Th2 or Th17 cells, was affected. Finally, RASF had a significantly lower IDO1 expression and a weaker Th cell suppressive capacity compared with osteoarthritis SF. We postulate that the suppression of Th cell growth by SF through tryptophan catabolism may play an important role in preventing inappropriate Th cell responses under normal conditions. However, expansion of Th17 cells that do not induce IDO1-mediated suppression and the reduced capacity of RASF to restrict Th cell proliferation through tryptophan metabolism may support the initiation and propagation of synovitis in RA patients.


Asunto(s)
Artritis Reumatoide/inmunología , Fibroblastos/inmunología , Indolamina-Pirrol 2,3,-Dioxigenasa/metabolismo , Células TH1/inmunología , Triptófano/metabolismo , Diferenciación Celular/inmunología , Cromatografía Líquida de Alta Presión , Técnicas de Cocultivo , Fibroblastos/metabolismo , Humanos , Immunoblotting , Indolamina-Pirrol 2,3,-Dioxigenasa/inmunología , Activación de Linfocitos/inmunología , Osteoartritis/inmunología , Reacción en Cadena de la Polimerasa , Membrana Sinovial/inmunología , Células Th17/inmunología , Células Th2/inmunología , Triptófano/inmunología
2.
Eur Heart J ; 39(38): 3521-3527, 2018 10 07.
Artículo en Inglés | MEDLINE | ID: mdl-29069365

RESUMEN

A large number of cardiovascular events are not prevented by current therapeutic regimens. In search for additional, innovative strategies, immune cells have been recognized as key players contributing to atherosclerotic plaque progression and destabilization. Particularly the role of innate immune cells is of major interest, following the recent paradigm shift that innate immunity, long considered to be incapable of learning, does exhibit immunological memory mediated via epigenetic reprogramming. Compelling evidence shows that atherosclerotic risk factors promote immune cell migration by pre-activation of circulating innate immune cells. Innate immune cell activation via metabolic and epigenetic reprogramming perpetuates a systemic low-grade inflammatory state in cardiovascular disease (CVD) that is also common in other chronic inflammatory disorders. This opens a new therapeutic area in which metabolic or epigenetic modulation of innate immune cells may result in decreased systemic chronic inflammation, alleviating CVD, and its co-morbidities.


Asunto(s)
Aterosclerosis/inmunología , Reprogramación Celular , Epigénesis Genética , Células Madre Hematopoyéticas/inmunología , Inmunidad Innata , Inflamación/inmunología , Monocitos/inmunología , Animales , Aterosclerosis/diagnóstico por imagen , Enfermedad Crónica , Humanos , Memoria Inmunológica , Inflamación/diagnóstico por imagen , Imagen Multimodal
3.
Ophthalmologica ; 238(4): 205-216, 2017.
Artículo en Inglés | MEDLINE | ID: mdl-28866675

RESUMEN

OBJECTIVE: To determine whether intraocular treatment with vascular endothelial growth factor (VEGF) inhibitors change systemic endothelial function (EF) in patients with neovascular age-related macular degeneration (AMD). METHODS: In this prospective, randomized, 2-center, double-masked controlled interventional trial, patients with neovascular and dry AMD were enrolled. Eligible neovascular AMD patients received 2 intravitreal loading doses of either ranibizumab 0.5 mg or bevacizumab 1.25 mg at 4-week intervals and were subsequently followed every 4 weeks and treated according to a pro re nata regime for up to 1 year. Patients with dry AMD served as controls. The primary endpoint was the change in EF assessed by flow-mediated dilatation (FMD) after 2 months of treatment with VEGF inhibitors in patients with AMD compared to patients with dry AMD. FMD was assessed with B-mode high-resolution ultrasonography of the left brachial artery. RESULTS: 24 patients with neovascular AMD and 26 patients with dry ADM were included in the trial. Treatment with VEGF inhibitors did not significantly change FMD (from 4.7 ± 2.4 to 3.9 ± 1.9% after 8 weeks, p = 0.07, and to 5.1 ± 2.0% after 1 year; p = 0.93 vs. baseline, respectively). CONCLUSIONS: EF did not significantly differ between patients with neovascular AMD treated with intravitreal VEGF inhibition and patients with dry AMD.


Asunto(s)
Bevacizumab/administración & dosificación , Ranibizumab/administración & dosificación , Factor A de Crecimiento Endotelial Vascular/antagonistas & inhibidores , Degeneración Macular Húmeda/tratamiento farmacológico , Anciano , Inhibidores de la Angiogénesis , Relación Dosis-Respuesta a Droga , Método Doble Ciego , Femenino , Estudios de Seguimiento , Humanos , Inyecciones Intravítreas , Mácula Lútea/patología , Masculino , Estudios Prospectivos , Tomografía de Coherencia Óptica , Agudeza Visual , Degeneración Macular Húmeda/diagnóstico
4.
J Immunol ; 193(10): 4945-51, 2014 Nov 15.
Artículo en Inglés | MEDLINE | ID: mdl-25320281

RESUMEN

In this study, we analyzed the methylation status of human promoters in rheumatoid arthritis synovial fibroblasts (RASF). Differentially methylated genes between RASF and osteoarthritis synovial fibroblasts (OASF) were identified by methylated DNA immunoprecipitation and hybridization to human promoter tiling arrays. The methylation status was confirmed by pyrosequencing. Gene and protein expression of differentially methylated genes was evaluated with real-time PCR, Western blot, and immunohistochemistry. Chromatin immunoprecipitation was used to measure the gene promoter-associated acetylation and methylation of histones. Transcription factor-specific targets were identified with microarray and luciferase assays. We found that the transcription factor T-box transcription factor 5 (TBX5) was less methylated in rheumatoid arthritis (RA) synovium and RASF than in osteoarthritis (OA) samples. Demethylation of the TBX5 promoter in RASF and RA synovium was accompanied by higher TBX5 expression than in OASF and OA synovium. In RA synovium, TBX5 expression was primarily localized to the synovial lining. In addition, the TBX5 locus was enriched in activating chromatin marks, such as histone 4 lysine 4 trimethylation and histone acetylation, in RASF. In our functional studies, we observed that 790 genes were differentially expressed by 2-6-fold after overexpression of TBX5 in OASF. Bioinformatic analysis of these genes revealed that the chemokines IL-8, CXCL12, and CCL20 were common targets of TBX5 in OASF. Taken together, our data show that TBX5 is a novel inducer of important chemokines in RASF. Thus, we conclude that RASF contribute to the inflammatory processes operating in the pathogenesis of RA via epigenetic control of TBX5.


Asunto(s)
Artritis Reumatoide/metabolismo , Epigénesis Genética , Fibroblastos/metabolismo , Membrana Sinovial/metabolismo , Proteínas de Dominio T Box/metabolismo , Acetilación , Artritis Reumatoide/inmunología , Artritis Reumatoide/patología , Quimiocina CCL20/genética , Quimiocina CCL20/inmunología , Quimiocina CCL20/metabolismo , Quimiocina CXCL12/genética , Quimiocina CXCL12/inmunología , Quimiocina CXCL12/metabolismo , Cromatina/inmunología , Cromatina/metabolismo , Biología Computacional , Fibroblastos/inmunología , Fibroblastos/patología , Humanos , Interleucina-8/genética , Interleucina-8/inmunología , Interleucina-8/metabolismo , Metilación , Regiones Promotoras Genéticas , Transducción de Señal , Membrana Sinovial/inmunología , Membrana Sinovial/patología , Proteínas de Dominio T Box/genética , Proteínas de Dominio T Box/inmunología , Transcripción Genética
5.
J Virol ; 88(2): 838-45, 2014 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-24173233

RESUMEN

Collagens, the most abundant proteins in animals, also occur in some recently described nucleocytoplasmic large DNA viruses such as Mimiviridae, which replicate in amoebae. To clarify the impact of viral collagens on the immune response of animals exposed to Mimiviridae, we have investigated the localization of collagens in Acanthamoeba polyphaga mimivirus particles and the response of mice to immunization with mimivirus particles. Using protein biotinylation, we have first shown that viral collagen encoded by open reading frame L71 is present at the surface of mimivirus particles. Exposure to mimivirus collagens elicited the production of anti-collagen antibodies in DBA/1 mice immunized intradermally with mimivirus protein extracts. This antibody response also targeted mouse collagen type II and was accompanied by T-cell reactivity to collagen and joint inflammation, as observed in collagen-induced arthritis following immunization of mice with bovine collagen type II. The broad distribution of nucleocytoplasmic large DNA viruses in the environment suggests that humans are constantly exposed to such large virus particles. A survey of blood sera from healthy human subjects and from rheumatoid arthritis patients indeed demonstrated that 30% of healthy-subject and 36% of rheumatoid arthritis sera recognized the major mimivirus capsid protein L425. Moreover, whereas 6% of healthy-subject sera recognized the mimivirus collagen protein L71, 22% of rheumatoid arthritis sera were positive for mimivirus L71. Accordingly, our study shows that environmental exposure to mimivirus represents a risk factor in triggering autoimmunity to collagens.


Asunto(s)
Artritis/virología , Colágeno/inmunología , Mimiviridae/inmunología , Proteínas Virales/inmunología , Animales , Anticuerpos Antivirales/inmunología , Artritis/inmunología , Bovinos , Colágeno/genética , Modelos Animales de Enfermedad , Humanos , Inmunización , Ratones , Ratones Endogámicos DBA , Mimiviridae/genética , Proteínas Virales/genética
6.
Rheumatology (Oxford) ; 53(8): 1520-6, 2014 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-24643522

RESUMEN

OBJECTIVES: S100A4 has been implicated in cancer and several inflammatory diseases, including RA. The aim of the present study was to determine whether S100A4 can stimulate proinflammatory cytokine production in mononuclear cells. METHODS: Peripheral blood mononuclear cells (PBMCs) isolated from patients with RA were stimulated with S100A4, S100A8, S100A9 and S100A12. The production of IL-1ß, IL-6 and TNF-α was measured by ELISA. Receptor for advanced glycation end products (RAGEs) and Toll-like receptor 4 (TLR4) signalling were examined. For signalling pathway blocking studies, inhibitors of myeloid differentiation primary response gene 88 (MyD88), nuclear factor kappa B (NF-κB) and the mitogen activated protein (MAP) kinases p38, extracellular signal-regulated kinase 1/2 (ERK1/2) and Jun N-terminal kinase (JNK) were used. MAP kinase activation was evaluated by western blotting. RESULTS: Stimulation of PBMCs with S100A4 significantly up-regulated IL-1ß, IL-6 and TNF-α production compared with unstimulated cells (P < 0.001). Importantly, the production of these cytokines was markedly enhanced in response to S100A4 compared with S100A8 and S100A12; however, it was less pronounced compared with S100A9. Furthermore, enhanced production of proinflammatory cytokines in S100A4-stimulated PMBCs was at least partly mediated via TLR4, but not RAGEs, and by activation of the transcription factor NF-κB and the MAP kinases p38 and ERK1/2. CONCLUSION: This is the first study to demonstrate that S100A4 can induce an inflammatory response mediated by TLR4 and by the activation of NF-κB and the kinases p38 and ERK1/2 in mononuclear cells from patients with RA. Therefore S100A4 may be a potential therapeutic target for immune-mediated diseases.


Asunto(s)
Artritis Reumatoide/metabolismo , Leucocitos Mononucleares/metabolismo , Proteínas S100/metabolismo , Transducción de Señal/fisiología , Receptor Toll-Like 4/metabolismo , Adulto , Anciano , Citocinas/metabolismo , Femenino , Humanos , Inflamación/metabolismo , Masculino , Persona de Mediana Edad , FN-kappa B/metabolismo , Proteína de Unión al Calcio S100A4 , Regulación hacia Arriba/fisiología
7.
Ann Rheum Dis ; 72(8): 1400-6, 2013 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-23234648

RESUMEN

OBJECTIVE: Fibrin deposits are characteristic of the synovial tissues in rheumatoid arthritis (RA). Once citrullinated, fibrin becomes an autoantigen and is thought to contribute in this way to perpetuate the disease. Our study aimed to analyse the responses of RA synovial fibroblasts (RASF) to native and citrullinated fibrin. METHODS: The transcriptome induced by fibrin in RASF was approached with whole-genome-based gene expression arrays. The upregulation of selected pro-inflammatory genes by fibrin was confirmed in additional primary cell cultures using quantitative PCR and ELISA. Citrullination reactions were carried out with recombinant human peptidylarginine deiminases (PAD) 2 and 4. RESULTS: In the whole-genome array native fibrin was found to modulate the gene expression profile of RASF, particularly upregulating mRNA levels of several pro-inflammatory cytokines. The induction of interleukin (IL)-6 and IL-8 by fibrin was confirmed in additional samples at both the mRNA and the protein level. Blocking and knockdown experiments showed the participation of toll-like receptor (TLR)4 in the induction of both cytokines. As compared with the native macromolecule, PAD2-citrullinated fibrin induced significantly higher expression of the pro-inflammatory cytokines in these cells. CONCLUSIONS: Our results suggest that fibrin mediates inflammatory responses in RASF via a TLR4 pathway. In this way, fibrin and particularly its citrullinated form may contribute to sustain the cytokine burst in RA.


Asunto(s)
Artritis Reumatoide/metabolismo , Fibrina/farmacología , Fibroblastos/efectos de los fármacos , Regulación de la Expresión Génica/efectos de los fármacos , Regulación hacia Arriba/efectos de los fármacos , Artritis Reumatoide/patología , Supervivencia Celular , Células Cultivadas , Citrulina/metabolismo , Citocinas/genética , Citocinas/metabolismo , Fibroblastos/metabolismo , Fibroblastos/patología , Expresión Génica , Perfilación de la Expresión Génica , Técnicas de Silenciamiento del Gen , Humanos , Hidrolasas/metabolismo , Análisis de Secuencia por Matrices de Oligonucleótidos , Arginina Deiminasa Proteína-Tipo 2 , Arginina Deiminasa Proteína-Tipo 4 , Desiminasas de la Arginina Proteica , ARN Mensajero/metabolismo , ARN Interferente Pequeño/farmacología , Proteínas Recombinantes , Membrana Sinovial/efectos de los fármacos , Membrana Sinovial/metabolismo , Membrana Sinovial/patología , Receptores Toll-Like/genética , Receptores Toll-Like/metabolismo
8.
Am J Pathol ; 180(5): 2144-55, 2012 May.
Artículo en Inglés | MEDLINE | ID: mdl-22449947

RESUMEN

A key feature of granulomatosis with polyangiitis (GPA; or Wegener's granulomatosis) is the granulomatous inflammation of the upper respiratory tract, which leads to the subsequent destruction of adjacent tissues. The aim of our work was to study the histopathological and cellular components of tissue destruction of human GPA tissue transplanted into immunodeficient mice. Biopsy specimens from patients with active GPA (n = 10) or sinusitis (controls, n = 6) were s.c. co-implanted with healthy allogeneic human nasal cartilage into immunodeficient pfp/rag2(-/-) mice. Transplants were examined for their destructive capability of the allografted human cartilage. In addition, nasal fibroblasts from patients with GPA (n = 8) and control healthy nasal fibroblasts (n = 5) were cultured, and cell proliferation and apoptosis were quantified. mRNA and protein levels of matrix metalloproteinases and cytokines were evaluated at baseline and after proinflammatory stimulation. GPA implants showed massive destruction of the co-implanted human cartilage, whereas cartilage destruction was only marginal in control samples. Destruction was mediated by human fibroblasts and could be inhibited by corticoid treatment. The up-regulated production of matrix metalloproteinases 1, 3, and 13 and cytokines IL-6 and IL-8 was found in vivo and in vitro. Although proliferation of isolated fibroblasts was comparable between GPA and controls, GPA samples showed a significant delay of apoptosis. The destruction of nasal cartilage in GPA is mainly mediated by fibroblasts that can be blocked by corticosteroids, and this tissue destruction is not dependent on the influx of leukocytes.


Asunto(s)
Fibroblastos/fisiología , Granulomatosis con Poliangitis/patología , Cartílagos Nasales/patología , Adulto , Anciano , Animales , Apoptosis/fisiología , Proliferación Celular , Células Cultivadas , Citocinas/biosíntesis , Dexametasona/farmacología , Dexametasona/uso terapéutico , Femenino , Fibroblastos/efectos de los fármacos , Fibroblastos/metabolismo , Perfilación de la Expresión Génica/métodos , Glucocorticoides/farmacología , Glucocorticoides/uso terapéutico , Granulomatosis con Poliangitis/complicaciones , Granulomatosis con Poliangitis/tratamiento farmacológico , Humanos , Tolerancia Inmunológica , Masculino , Metaloproteinasas de la Matriz/biosíntesis , Ratones , Ratones Noqueados , Persona de Mediana Edad , Cartílagos Nasales/trasplante , Mucosa Nasal/patología , Mucosa Nasal/trasplante , Deformidades Adquiridas Nasales/etiología , Deformidades Adquiridas Nasales/patología , ARN Mensajero/genética , Reacción en Cadena en Tiempo Real de la Polimerasa/métodos , Adulto Joven
9.
Arthritis Rheum ; 64(6): 1809-17, 2012 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-22170508

RESUMEN

OBJECTIVE: Global DNA hypomethylation in rheumatoid arthritis synovial fibroblasts (RASFs) contributes to their intrinsic activation. The aim of this study was to investigate whether increased polyamine metabolism is associated with a decreased level of S-adenosyl methionine (SAM), causing global DNA hypomethylation. METHODS: Synovial fibroblasts were isolated from synovial tissue obtained from 12 patients with RA and from 6 patients with osteoarthritis (OA). The cells were stained for S-adenosyl methionine decarboxylase (AMD), spermidine/spermine N1-acetyltransferase (SSAT1), polyamine-modulated factor 1-binding protein 1 (PMFBP1), solute carrier family 3 member 2 (SLC3A2), DNA methyltransferase 1 (DNMT-1), α9 integrin, and ß1 integrin and analyzed by flow cytometry. Nuclear 5-methylcytosine (5-MeC) was measured by flow cytometry, the expression of diacetylspermine (DASp) in cell culture supernatants and cell extracts was determined by enzyme-linked immunosorbent assay, and SAM expression in cell extracts was measured by fluorometry. RESULTS: The expression of SSAT1, AMD, and PMFBP1 was significantly increased in RASFs compared with OASFs. The expression of DASp in cell culture supernatants and the expression of SLC3A2 were significantly elevated in RASFs. The levels of SAM in cell culture extracts, as well as the levels of DNMT-1 protein and 5-MeC, were significantly reduced in RASFs. Parameters of polyamine metabolism were negatively correlated with the expression of SAM, DNMT-1, and 5-MeC. CONCLUSION: These data clearly show that intrinsic elevations of PMFBP1 and SSAT1 enhance the catabolism and recycling of polyamines in RASFs and suggest that high consumption of SAM via this pathway is an important factor contributing to global DNA hypomethylation in these cells.


Asunto(s)
Artritis Reumatoide/metabolismo , Metilación de ADN , Fibroblastos/metabolismo , Poliaminas/metabolismo , Membrana Sinovial/metabolismo , Anciano , Artritis Reumatoide/genética , Artritis Reumatoide/patología , Células Cultivadas , ADN/genética , ADN/metabolismo , Femenino , Fibroblastos/patología , Citometría de Flujo , Expresión Génica , Humanos , Masculino , Persona de Mediana Edad , Membrana Sinovial/patología
10.
Arthritis Rheum ; 64(6): 1771-9, 2012 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-22161761

RESUMEN

OBJECTIVE: To investigate the expression and effect of the microRNA-34 (miR-34) family on apoptosis in rheumatoid arthritis synovial fibroblasts (RASFs). METHODS: Expression of the miR-34 family in synovial fibroblasts with or without stimulation with Toll-like receptor (TLR) ligands, tumor necrosis factor α (TNFα), interleukin-1ß (IL-1ß), hypoxia, or 5-azacytidine was analyzed by real-time polymerase chain reaction (PCR). Promoter methylation was studied by combined bisulfite restriction analysis. The effects of overexpression and silencing of miR-34a and miR-34a* on apoptosis were analyzed by annexin V/propidium iodide staining. Production of X-linked inhibitor of apoptosis protein (XIAP) was assessed by real-time PCR and immunohistochemistry analysis. Reporter gene assay was used to study the signaling pathways of miR-34a*. RESULTS: Basal expression levels of miR-34a* were found to be reduced in synovial fibroblasts from RA patients compared to osteoarthritis patients, whereas levels of miR-34a, miR-34b/b*, and miR-34c/c* did not differ. Neither TNFα, IL-1ß, TLR ligands, nor hypoxia altered miR-34a* expression. However, we demonstrated that the promoter of miR-34a/34a* was methylated and showed that transcription of the miR-34a duplex was induced upon treatment with demethylating agents. Enforced expression of miR-34a* led to an increased rate of FasL- and TRAIL-mediated apoptosis in RASFs. Moreover, levels of miR-34a* were highly correlated with expression of XIAP, which was found to be up-regulated in RA synovial cells. Finally, we identified XIAP as a direct target of miR-34a*. CONCLUSION: Our data provide evidence of a methylation-specific down-regulation of proapoptotic miR-34a* in RASFs. Decreased expression of miR- 34a* results in up-regulation of its direct target XIAP, thereby contributing to resistance of RASFs to apoptosis.


Asunto(s)
Apoptosis/fisiología , Artritis Reumatoide/metabolismo , Regulación hacia Abajo , Fibroblastos/metabolismo , MicroARNs/metabolismo , Membrana Sinovial/metabolismo , Artritis Reumatoide/genética , Artritis Reumatoide/patología , Azacitidina/farmacología , Células Cultivadas , Fibroblastos/patología , Humanos , Hipoxia/metabolismo , Hipoxia/patología , Interleucina-1beta/farmacología , MicroARNs/genética , Membrana Sinovial/patología , Factor de Necrosis Tumoral alfa/farmacología , Proteína Inhibidora de la Apoptosis Ligada a X/genética , Proteína Inhibidora de la Apoptosis Ligada a X/metabolismo
11.
Eur Heart J ; 33(13): 1589-97, 2012 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-22240497

RESUMEN

AIMS: Extracts from pine tree bark containing a variety of flavonoids have been used in traditional medicine. Pycnogenol is a proprietary bark extract of the French maritime pine tree (Pinus pinaster ssp. atlantica) that exerts antioxidative, anti-inflammatory, and anti-platelet effects. However, the effects of Pycnogenol on endothelial dysfunction, a precursor of atherosclerosis and cardiovascular events, remain still elusive. METHODS AND RESULTS: Twenty-three patients with coronary artery disease (CAD) completed this randomized, double-blind, placebo-controlled cross-over study. Patients received Pycnogenol (200 mg/day) for 8 weeks followed by placebo or vice versa on top of standard cardiovascular therapy. Between the two treatment periods, a 2-week washout period was scheduled. At baseline and after each treatment period, endothelial function, non-invasively assessed by flow-mediated dilatation (FMD) of the brachial artery using high-resolution ultrasound, biomarkers of oxidative stress and inflammation, platelet adhesion, and 24 h blood pressure monitoring were evaluated. In CAD patients, Pycnogenol treatment was associated with an improvement of FMD from 5.3 ± 2.6 to 7.0 ± 3.1 (P < 0.0001), while no change was observed with placebo (5.4 ± 2.4 to 4.7 ± 2.0; P = 0.051). This difference between study groups was significant [estimated treatment effect 2.75; 95% confidence interval (CI): 1.75, 3.75, P < 0.0001]. 15-F(2t)-Isoprostane, an index of oxidative stress, significantly decreased from 0.71 ± 0.09 to 0.66 ± 0.13 after Pycnogenol treatment, while no change was observed in the placebo group (mean difference 0.06 pg/mL with an associated 95% CI (0.01, 0.11), P = 0.012]. Inflammation markers, platelet adhesion, and blood pressure did not change after treatment with Pycnogenol or placebo. CONCLUSION: This study provides the first evidence that the antioxidant Pycnogenol improves endothelial function in patients with CAD by reducing oxidative stress.


Asunto(s)
Antioxidantes/administración & dosificación , Enfermedad de la Arteria Coronaria/tratamiento farmacológico , Endotelio Vascular/efectos de los fármacos , Flavonoides/administración & dosificación , Vasodilatación/efectos de los fármacos , Adulto , Anciano , Antiinflamatorios/administración & dosificación , Antihipertensivos/administración & dosificación , Arginina/análogos & derivados , Arginina/metabolismo , Biomarcadores/metabolismo , Plaquetas/efectos de los fármacos , Presión Sanguínea/efectos de los fármacos , Proteína C-Reactiva/metabolismo , Enfermedad de la Arteria Coronaria/fisiopatología , Estudios Cruzados , Método Doble Ciego , Endotelina-1/efectos de los fármacos , Femenino , Humanos , Masculino , Persona de Mediana Edad , Estrés Oxidativo/efectos de los fármacos , Extractos Vegetales , Estudios Prospectivos , Resistencia al Corte , Resultado del Tratamiento , Vasodilatadores/administración & dosificación
12.
Circulation ; 122(18): 1789-96, 2010 Nov 02.
Artículo en Inglés | MEDLINE | ID: mdl-20956208

RESUMEN

BACKGROUND: Because traditional nonsteroidal antiinflammatory drugs are associated with increased risk for acute cardiovascular events, current guidelines recommend acetaminophen as the first-line analgesic of choice on the assumption of its greater cardiovascular safety. Data from randomized clinical trials prospectively addressing cardiovascular safety of acetaminophen, however, are still lacking, particularly in patients at increased cardiovascular risk. Hence, the aim of this study was to evaluate the safety of acetaminophen in patients with coronary artery disease. METHODS AND RESULTS: The 33 patients with coronary artery disease included in this randomized, double-blind, placebo-controlled, crossover study received acetaminophen (1 g TID) on top of standard cardiovascular therapy for 2 weeks. Ambulatory blood pressure, heart rate, endothelium-dependent and -independent vasodilatation, platelet function, endothelial progenitor cells, markers of the renin-angiotensin system, inflammation, and oxidative stress were determined at baseline and after each treatment period. Treatment with acetaminophen resulted in a significant increase in mean systolic (from 122.4±11.9 to 125.3±12.0 mm Hg P=0.02 versus placebo) and diastolic (from 73.2±6.9 to 75.4±7.9 mm Hg P=0.02 versus placebo) ambulatory blood pressures. On the other hand, heart rate, endothelial function, early endothelial progenitor cells, and platelet function did not change. CONCLUSIONS: This study demonstrates for the first time that acetaminophen induces a significant increase in ambulatory blood pressure in patients with coronary artery disease. Thus, the use of acetaminophen should be evaluated as rigorously as traditional nonsteroidal antiinflammatory drugs and cyclooxygenase-2 inhibitors, particularly in patients at increased cardiovascular risk. CLINICAL TRIAL REGISTRATION: URL: http://www.clinicaltrials.gov. Unique identifier: NCT00534651.


Asunto(s)
Acetaminofén/efectos adversos , Acetaminofén/farmacología , Antiinflamatorios no Esteroideos/efectos adversos , Antiinflamatorios no Esteroideos/farmacología , Presión Sanguínea/efectos de los fármacos , Enfermedad de la Arteria Coronaria/fisiopatología , Hipertensión/inducido químicamente , Anciano , Presión Sanguínea/fisiología , Ritmo Circadiano/efectos de los fármacos , Ritmo Circadiano/fisiología , Estudios Cruzados , Relación Dosis-Respuesta a Droga , Método Doble Ciego , Endotelio Vascular/efectos de los fármacos , Endotelio Vascular/fisiopatología , Femenino , Frecuencia Cardíaca/efectos de los fármacos , Frecuencia Cardíaca/fisiología , Humanos , Hipertensión/epidemiología , Hipertensión/fisiopatología , Masculino , Persona de Mediana Edad , Adhesividad Plaquetaria/efectos de los fármacos , Adhesividad Plaquetaria/fisiología , Estudios Prospectivos , Factores de Riesgo , Vasodilatación/efectos de los fármacos , Vasodilatación/fisiología
13.
Rheumatology (Oxford) ; 50(10): 1766-72, 2011 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-21712367

RESUMEN

OBJECTIVES: The S100A4 protein is known as a metastasis promoting factor; however, its involvement in non-malignant diseases such as RA and psoriasis has been recently described. The aim of this study was to investigate the expression and possible role of S100A4 in idiopathic inflammatory myopathies. METHODS: S100A4 protein expression was detected by immunohistochemistry in muscle tissue from control individuals (n = 11) and patients with PM and DM (n = 8/6). IF staining was used to co-localize S100A4 with selected cells. Cytokine expression and protein synthesis in S100A4-treated cells were analysed by RT-PCR and ELISA. RESULTS: S100A4 protein was significantly up-regulated in muscle tissue of patients with inflammatory myopathies compared with control individuals and was associated particularly with the presence of mononuclear infiltrates. Only few regenerating muscle fibres in PM/DM expressed S100A4. Then we analysed the effect of S100A4 on human myocytes and peripheral blood mononuclear cells (PBMCs). Although S100A4 did not affect myocytes, stimulation of PBMCs with S100A4 significantly induced the expression and synthesis of TNF-α, IL-1ß and IL-6, but not of IFN-α. We showed that S100A4 is not directly involved in perforin/granzyme B-induced apoptosis and that it does not modulate the expression of Bax and Bcl2 mRNA in myocytes and PBMCs. CONCLUSION: Increased expression of S100A4 in inflamed muscle tissue highlights its potential role in the pathogenesis of inflammatory myopathies. S100A4 may act as a cytokine-like factor indirectly promoting muscle fibre damage by stimulating mononuclear cells to increase the synthesis of pro-inflammatory cytokines.


Asunto(s)
Dermatomiositis/metabolismo , Polimiositis/metabolismo , Proteínas S100/metabolismo , Adulto , Anciano , Biomarcadores/metabolismo , Citocinas/genética , Citocinas/metabolismo , Dermatomiositis/genética , Dermatomiositis/patología , Femenino , Expresión Génica , Humanos , Leucocitos Mononucleares/efectos de los fármacos , Leucocitos Mononucleares/metabolismo , Lipopolisacáridos/farmacología , Masculino , Persona de Mediana Edad , Fibras Musculares Esqueléticas/efectos de los fármacos , Fibras Musculares Esqueléticas/metabolismo , Fibras Musculares Esqueléticas/patología , Músculo Esquelético/metabolismo , Músculo Esquelético/patología , Polimiositis/genética , Polimiositis/patología , ARN Mensajero/metabolismo , Proteínas Recombinantes/farmacología , Proteína de Unión al Calcio S100A4 , Proteínas S100/genética , Proteínas S100/farmacología , Regulación hacia Arriba/efectos de los fármacos
14.
Arthritis Rheum ; 62(9): 2673-9, 2010 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-20496420

RESUMEN

OBJECTIVE: To explore whether the increased expression of long interspersed nuclear element 1 (LINE-1; L1) messenger RNA (mRNA) and protein in rheumatoid arthritis synovial fibroblasts (RASFs) is associated with decreased expression of Trex-1, an exonuclease involved in the metabolization of L1 DNA:RNA hybrids. METHODS: Chromatin immunoprecipitation was used to detect L1-related p40 protein (L1-ORF1p) binding sequences in RASFs. Luciferase activity was measured in the synovial fibroblasts following cotransfection of the episomal plasmid with pJM105 expressing L1-ORF1p and pGL3-TS3 carrying the target sequence for L1-ORF1p. This luciferase reporter assay was used to compare the activity between RASFs and osteoarthritis synovial fibroblasts (OASFs) and to assess correlations of luciferase activity with the expression of Trex-1 measured by flow cytometry. The expression of Trex-1 mRNA and protein was also compared using real-time polymerase chain reaction, immunohistochemistry, and Western blot analyses. The role of Trex-1 in the L1-ORF1p-mediated luciferase activity assay was studied using interfering RNAs (iRNA) and a Trex-1 expression vector. RESULTS: Increased luciferase activity occurred after cotransfection of synovial fibroblasts with pJM105 and pGL3-TS3. L1-ORF1p activity was increased in RASFs as compared with OASFs, and this was correlated inversely with the expression of Trex-1. Levels of Trex-1 mRNA and protein were lower in RASFs than in OASFs. After transfection of the L1 expression plasmid, Trex-1 mRNA levels increased in OASFs, but not in RASFs. The addition of iRNA against Trex-1, however, resulted in an enhancement of L1-ORF1p activity in OASFs to the levels measured in RASFs. Overexpression of Trex-1 inhibited 5-azacytidine-induced expression of p38δ MAPK, a gene carrying the TS3 sequence. CONCLUSION: The deficiency of Trex-1 in RASFs allows a longer half-life of gene products encoded by active endogenous L1 retrotransposons. This pathway may play a role in diseases in which the cells exhibit a "spontaneous" aggressive behavior.


Asunto(s)
Artritis Reumatoide/metabolismo , Exodesoxirribonucleasas/metabolismo , Fibroblastos/metabolismo , Elementos de Nucleótido Esparcido Largo/genética , Osteoartritis/metabolismo , Fosfoproteínas/metabolismo , Membrana Sinovial/metabolismo , Artritis Reumatoide/patología , Azacitidina/farmacología , Células Cultivadas , Regulación hacia Abajo , Exodesoxirribonucleasas/genética , Fibroblastos/efectos de los fármacos , Fibroblastos/patología , Citometría de Flujo , Expresión Génica , Silenciador del Gen , Humanos , Inmunohistoquímica , Análisis de Secuencia por Matrices de Oligonucleótidos , Osteoartritis/patología , Fosfoproteínas/genética , ARN Mensajero/metabolismo , ARN Interferente Pequeño/genética , ARN Interferente Pequeño/metabolismo , Membrana Sinovial/efectos de los fármacos , Membrana Sinovial/patología , Transfección/métodos
15.
Adv Exp Med Biol ; 711: 137-49, 2011.
Artículo en Inglés | MEDLINE | ID: mdl-21627047

RESUMEN

In this chapter, we discuss the current understanding of the possible epigenetics changes that occur in rheumatoid arthritis. In particular, we describe that deregulation ofDNA methylation and histone modifications can occur in the immune system and lead to rheumatoid arthritis. In addition, we discuss the role of rheumatoid arthritis synovial fibroblasts in autoimmunity. Examples of changes in DNA methylation and histone modification occurring in synovial fibroblasts during the disease process are reviewed in this chapter. In conclusion, we discuss the possible use of epigenetic therapy and describe future experiments that can elucidate further the epigenetic changes observed in the disease.


Asunto(s)
Artritis Reumatoide/genética , Epigénesis Genética , Envejecimiento , Animales , Artritis Reumatoide/etiología , Artritis Reumatoide/inmunología , Autoinmunidad , Ambiente , Fibroblastos/metabolismo , Humanos , Membrana Sinovial/citología , Membrana Sinovial/metabolismo , Inactivación del Cromosoma X
16.
Eur Heart J ; 31(12): 1457-69, 2010 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-20447947

RESUMEN

AIMS: Inflammation plays a key role in acute coronary syndromes (ACS). Toll-like receptors (TLR) on leucocytes mediate inflammation and immune responses. We characterized leucocytes and TLR expression within coronary thrombi and compared cytokine levels from the site of coronary occlusion with aortic blood (AB) in ACS patients. METHODS AND RESULTS: In 18 ACS patients, thrombi were collected by aspiration during primary percutaneous coronary intervention. Thrombi and AB from these patients as well as AB from 10 age-matched controls without coronary artery disease were assessed by FACS analysis for cellular distribution and TLR expression. For further discrimination of ACS specificity, seven non-coronary intravascular thrombi and eight thrombi generated in vitro were analysed. In 17 additional patients, cytokine levels were determined in blood samples from the site of coronary occlusion under distal occlusion and compared with AB. In coronary thrombi from ACS, the percentage of monocytes related to the total leucocyte count was greater than in AB (47 vs. 20%, P = 0.0002). In thrombi, TLR-4 and TLR-2 were overexpressed on CD14-labelled monocytes, and TLR-2 was increased on CD66b-labelled granulocytes, in comparison with leucocytes in AB. In contrast, in vitro and non-coronary thrombi exhibited no overexpression of TLR-4. Local blood samples taken under distal occlusion revealed elevated concentrations of chemokines (IL-8, MCP-1, eotaxin, MIP-1alpha, and IP-10) and cytokines (IL-1ra, IL-6, IL-7, IL-12, IL-17, IFN-alpha, and granulocyte-macrophage colony-stimulating factor) regulating both innate and adaptive immunity (all P < 0.05). CONCLUSION: In ACS patients, monocytes accumulate within thrombi and specifically overexpress TLR-4. Together with the local expression patterns of chemokines and cytokines, the increase of TLR-4 reflects a concerted activation of this inflammatory pathway at the site of coronary occlusion in ACS.


Asunto(s)
Síndrome Coronario Agudo/metabolismo , Oclusión Coronaria/metabolismo , Trombosis Coronaria/metabolismo , Citocinas/metabolismo , Monocitos/metabolismo , Receptores Toll-Like/metabolismo , Síndrome Coronario Agudo/patología , Anciano , Aorta , Estudios de Casos y Controles , Trombosis Coronaria/patología , Femenino , Humanos , Inmunohistoquímica , Recuento de Leucocitos , Masculino , Persona de Mediana Edad , Factores de Riesgo
17.
Circulation ; 120(5): 391-9, 2009 Aug 04.
Artículo en Inglés | MEDLINE | ID: mdl-19620501

RESUMEN

BACKGROUND: Intravascular ultrasound of drug-eluting stent (DES) thrombosis (ST) reveals a high incidence of incomplete stent apposition (ISA) and vessel remodeling. Autopsy specimens of DES ST show delayed healing and hypersensitivity reactions. The present study sought to correlate histopathology of thrombus aspirates with intravascular ultrasound findings in patients with very late DES ST. METHODS AND RESULTS: The study population consisted of 54 patients (28 patients with very late DES ST and 26 controls). Of 28 patients with very late DES ST, 10 patients (1020+/-283 days after implantation) with 11 ST segments (5 sirolimus-eluting stents, 5 paclitaxel-eluting stents, 1 zotarolimus-eluting stent) underwent both thrombus aspiration and intravascular ultrasound investigation. ISA was present in 73% of cases with an ISA cross-sectional area of 6.2+/-2.4 mm(2) and evidence of vessel remodeling (index, 1.6+/-0.3). Histopathological analysis showed pieces of fresh thrombus with inflammatory cell infiltrates (DES, 263+/-149 white blood cells per high-power field) and eosinophils (DES, 20+/-24 eosinophils per high-power field; sirolimus-eluting stents, 34+/-28; paclitaxel-eluting stents, 6+/-6; P for sirolimus-eluting stents versus paclitaxel-eluting stents=0.09). The mean number of eosinophils per high-power field was higher in specimens from very late DES ST (20+/-24) than in those from spontaneous acute myocardial infarction (7+/-10), early bare-metal stent ST (1+/-1), early DES ST (1+/-2), and late bare-metal stent ST (2+/-3; P from ANOVA=0.038). Eosinophil count correlated with ISA cross-sectional area, with an average increase of 5.4 eosinophils per high-power field per 1-mm(2) increase in ISA cross-sectional area. CONCLUSIONS: Very late DES thrombosis is associated with histopathological signs of inflammation and intravascular ultrasound evidence of vessel remodeling. Compared with other causes of myocardial infarction, eosinophilic infiltrates are more common in thrombi harvested from very late DES thrombosis, particularly in sirolimus-eluting stents, and correlate with the extent of stent malapposition.


Asunto(s)
Enfermedad de la Arteria Coronaria/terapia , Trombosis Coronaria , Stents Liberadores de Fármacos/efectos adversos , Vasculitis , Anciano , Angioplastia Coronaria con Balón , Biomarcadores/metabolismo , Trombosis Coronaria/diagnóstico por imagen , Trombosis Coronaria/inmunología , Trombosis Coronaria/patología , Vasos Coronarios/diagnóstico por imagen , Vasos Coronarios/inmunología , Vasos Coronarios/patología , Eosinófilos/patología , Femenino , Humanos , Masculino , Persona de Mediana Edad , Trombectomía , Ultrasonografía Intervencional , Vasculitis/diagnóstico por imagen , Vasculitis/etiología , Vasculitis/patología
18.
Ann Rheum Dis ; 69(5): 898-902, 2010 May.
Artículo en Inglés | MEDLINE | ID: mdl-19497915

RESUMEN

OBJECTIVE: To evaluate the decrease of cartilage destruction by a novel orally active and specific matrix metalloproteinase 13 (MMP-13) inhibitor in three different animal models of rheumatoid arthritis (RA). MATERIALS AND METHODS: The SCID mouse co-implantation model of RA, the collagen-induced arthritis (CIA) model in mice and the antigen-induced arthritis model (AIA) in rabbits were used. RESULTS: In the SCID mouse co-implantation model, the MMP-13 inhibitor reduced cartilage destruction by 75%. In the CIA model of RA, the MMP-13 inhibitor resulted in a significant and dose-dependent decrease in clinical symptoms as well as of cartilage erosion by 38% (30 mg/kg), 28% (10 mg/kg) and 21% (3 mg/kg). No significant effects were seen in the AIA model. No toxic effects were seen in all three animal models. CONCLUSION: Although several MMPs in concert with other proteinases have a role in the process of cartilage destruction, there is a need for highly selective MMP inhibitors to reduce severe side effects that occur with non-specific inhibitors. Significant inhibition of MMP-13 reduced cartilage erosions in two of three tested animal models of RA. These results strongly support the development of this class of drugs to reduce or halt joint destruction in patients with RA.


Asunto(s)
Antirreumáticos/uso terapéutico , Artritis Experimental/tratamiento farmacológico , Artritis Reumatoide/tratamiento farmacológico , Inhibidores Enzimáticos/uso terapéutico , Inhibidores de la Metaloproteinasa de la Matriz , Administración Oral , Animales , Antirreumáticos/administración & dosificación , Artritis Experimental/enzimología , Artritis Experimental/patología , Artritis Reumatoide/enzimología , Artritis Reumatoide/patología , Cartílago Articular/patología , Modelos Animales de Enfermedad , Relación Dosis-Respuesta a Droga , Evaluación Preclínica de Medicamentos/métodos , Inhibidores Enzimáticos/administración & dosificación , Femenino , Humanos , Masculino , Metaloproteinasa 1 de la Matriz/metabolismo , Metaloproteinasa 13 de la Matriz/metabolismo , Ratones , Ratones SCID , Conejos , Membrana Sinovial/enzimología
19.
J Autoimmun ; 35(1): 15-22, 2010 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-20079608

RESUMEN

The aggressive phenotype of RA synovial fibroblasts (RASF) is characterised by the increased expression of matrix metalloproteinase (MMP)-1 as well as the small ubiquitin like modifier (SUMO)-1 and decreased expression of SUMO-specific protease SENP1. Since we showed an increased activity of acetyltransferases in this autoimmune disease, we wanted to analyze whether this affects the expression of MMP-1 and can be reversed by the reconstitution of SENP1. In RASF, the acetylation of histone H4 was significantly increased in the distal region of the MMP-1 promoter by 274 +/- 36% compared to OASF. Most interestingly, overexpression of SENP1 in RASF decreased acetylation specifically in this region by 51 +/- 0.5% and globally by 73 +/- 11%. Furthermore, the overexpression of SENP1 resulted in a downregulation of MMP-1 at both the mRNA (58 +/- 7%) and protein levels (28 +/- 6%), significantly reduced the invasiveness of RASF (from 34 +/- 9% to 2 +/- 2%) and led to an accumulation of histone deacetylase 4 (HDAC4) on the MMP-1 promoter (197 +/- 36%). Interestingly, SENP1 failed to modulate the expression of MMP-1 in the cells silenced for HDAC4. This is the first study linking the SUMOylation pathway and the production of MMP-1 to an epigenetic control mechanism mediated through histone acetylation which has a functional consequence for the invasiveness of RASF.


Asunto(s)
Artritis Reumatoide/genética , Endopeptidasas/metabolismo , Fibroblastos/metabolismo , Metaloproteinasa 1 de la Matriz/metabolismo , Invasividad Neoplásica/genética , Acetilación , Animales , Artritis Reumatoide/metabolismo , Línea Celular , Clonación Molecular , Cisteína Endopeptidasas , Perros , Endopeptidasas/genética , Endopeptidasas/inmunología , Epigénesis Genética , Fibroblastos/inmunología , Fibroblastos/patología , Histona Desacetilasas/genética , Histona Desacetilasas/metabolismo , Histonas/genética , Histonas/metabolismo , Humanos , Metaloproteinasa 1 de la Matriz/genética , Metaloproteinasa 1 de la Matriz/inmunología , Regiones Promotoras Genéticas , ARN Interferente Pequeño/genética , Proteínas Represoras/genética , Proteínas Represoras/metabolismo , Membrana Sinovial/patología , Transgenes/genética
20.
Circulation ; 117(17): 2262-9, 2008 Apr 29.
Artículo en Inglés | MEDLINE | ID: mdl-18427133

RESUMEN

BACKGROUND: The excess in cardiovascular risk in patients with rheumatoid arthritis provides a strong rationale for early therapeutical interventions. In view of the similarities between atherosclerosis and rheumatoid arthritis and the proven benefit of angiotensin-converting enzyme inhibitors in atherosclerotic vascular disease, it was the aim of the present study to delineate the impact of ramipril on endothelial function as well as on markers of inflammation and oxidative stress in patients with rheumatoid arthritis. METHODS AND RESULTS: Eleven patients with rheumatoid arthritis were included in this randomized, double-blind, crossover study to receive ramipril in an uptitration design (2.5 to 10 mg) for 8 weeks followed by placebo, or vice versa, on top of standard antiinflammatory therapy. Endothelial function assessed by flow-mediated dilation of the brachial artery, markers of inflammation and oxidative stress, and disease activity were investigated at baseline and after each treatment period. Endothelial function assessed by flow-mediated dilation increased from 2.85+/-1.49% to 4.00+/-1.81% (P=0.017) after 8 weeks of therapy with ramipril but did not change with placebo (from 2.85+/-1.49% to 2.84+/-2.47%; P=0.88). Although systolic blood pressure and heart rate remained unaltered, diastolic blood pressure decreased slightly from 78+/-7 to 74+/-6 mm Hg (P=0.03). Tumor necrosis factor-alpha showed a significant inverse correlation with flow-mediated dilation (r=-0.408, P=0.02), and CD40 significantly decreased after ramipril therapy (P=0.049). CONCLUSIONS: Angiotensin-converting enzyme inhibition with 10 mg/d ramipril for 8 weeks on top of current antiinflammatory treatment markedly improved endothelial function in patients with rheumatoid arthritis. This finding suggests that angiotensin-converting enzyme inhibition may provide a novel strategy to prevent cardiovascular events in these patients.


Asunto(s)
Inhibidores de la Enzima Convertidora de Angiotensina/administración & dosificación , Artritis Reumatoide/complicaciones , Aterosclerosis/prevención & control , Ramipril/administración & dosificación , Vasculitis/prevención & control , Adulto , Anciano , Artritis Reumatoide/epidemiología , Aterosclerosis/epidemiología , Biomarcadores , Células Cultivadas , Estudios Cruzados , Células Endoteliales/citología , Células Endoteliales/efectos de los fármacos , Células Endoteliales/fisiología , Femenino , Humanos , Masculino , Persona de Mediana Edad , Análisis de Secuencia por Matrices de Oligonucleótidos , Estrés Oxidativo/efectos de los fármacos , Sistema Renina-Angiotensina/efectos de los fármacos , Factores de Riesgo , Resultado del Tratamiento , Venas Umbilicales/citología , Vasculitis/epidemiología , Vasodilatación/efectos de los fármacos
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