8 results on '"Angela Amoruso"'
Search Results
2. Modulation of human monocyte/macrophage activity by tocilizumab, abatacept and etanercept: An in vitro study
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Angela Amoruso, Sandra Brunelleschi, Daniele Sola, L. Fresu, Joyce Afrakoma Obeng, and Gian Luca Ermanno Camaschella
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musculoskeletal diseases ,0301 basic medicine ,Cell Survival ,Peroxisome proliferator-activated receptor ,Inflammation ,Pharmacology ,Antibodies, Monoclonal, Humanized ,Gene Expression Regulation, Enzymologic ,Monocytes ,Etanercept ,Abatacept ,03 medical and health sciences ,chemistry.chemical_compound ,Tocilizumab ,medicine ,Humans ,skin and connective tissue diseases ,Receptor ,chemistry.chemical_classification ,business.industry ,Macrophages ,medicine.disease ,PPAR gamma ,Phenotype ,030104 developmental biology ,Matrix Metalloproteinase 9 ,chemistry ,Mechanism of action ,Rheumatoid arthritis ,Immunology ,medicine.symptom ,business ,Biomarkers ,medicine.drug - Abstract
Tocilizumab, etanercept and abatacept are biological drugs used in the therapy of Rheumatoid Arthritis (RA). Their mechanism of action is well documented but their direct effects on human monocytes/macrophages have not been fully investigated. The objective of this study was to evaluate in vitro the influence of these drugs on monocytes/macrophages from healthy volunteers. Human monocytes were isolated from healthy anonymous volunteers and cultured as such or differentiated to monocyte-derived macrophages (MDMs). The effect of tocilizumab, etanercept and abatacept (at concentrations similar to those in plasma of patients) on superoxide anion production, matrix metalloproteinase-9 (MMP-9) gene expression and activity, Peroxisome Proliferator-Activated Receptor (PPAR)γ expression and cell phenotype was evaluated. Exposure of monocytes/macrophages to tocilizumab, etanercept or abatacept resulted in a significant decrease of the PMA-induced superoxide anion production. Interestingly, the expression of PPARγ was significantly increased only by tocilizumab, while etanercept was the only one able to significantly reduce MMP-9 gene expression and inhibit the LPS-induced MMP-9 activity in monocytes. When etanercept and abatacept were added to the differentiating medium, both significantly reduced the amount of CD206(+)MDM. This study demonstrates that etanercept, abatacept and tocilizumab affect differently human monocytes/macrophages. In particular, the IL-6 antagonist tocilizumab seems to be more effective in inducing an anti-inflammatory phenotype of monocytes/macrophages compared to etanercept and abatacept, also in light of the up-regulation of PPARγ whose anti-inflammatory effects are well recognised.
- Published
- 2016
3. Particulate matter induces prothrombotic microparticle shedding by human mononuclear and endothelial cells
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Silvia Petrini, Laura Pergoli, Valentina Bollati, Angela Amoruso, Roberto Pedrinelli, Lotte Gravendonk, Valentina Scalise, Pierluigi Paggiaro, Cristina Balia, Alessandro Celi, and Tommaso Neri
- Subjects
0301 basic medicine ,Endothelial cells ,Inflammation ,Phosphatidylserines ,Microparticles ,030204 cardiovascular system & hematology ,Toxicology ,Peripheral blood mononuclear cell ,Calcium in biology ,Cell-Derived Microparticles ,03 medical and health sciences ,chemistry.chemical_compound ,Tissue factor ,0302 clinical medicine ,Human Umbilical Vein Endothelial Cells ,medicine ,Humans ,Microparticle ,Cells, Cultured ,Air Pollutants ,Thrombosis ,General Medicine ,Phosphatidylserine ,Particulates ,Cell biology ,030104 developmental biology ,Mononuclear cells ,chemistry ,Immunology ,Leukocytes, Mononuclear ,Particulate matter ,Calcium ,medicine.symptom - Abstract
Particulate airborne pollution is associated with increased cardiopulmonary morbidity. Microparticles are extracellular vesicles shed by cells upon activation or apoptosis involved in physiological processes such as coagulation and inflammation, including airway inflammation. We investigated the hypothesis that particulate matter causes the shedding of microparticles by human mononuclear and endothelial cells. Cells, isolated from the blood and the umbilical cords of normal donors, were cultured in the presence of particulate from a standard reference. Microparticles were assessed in the supernatant as phosphatidylserine concentration. Microparticle-associated tissue factor was assessed by an one-stage clotting assay. Nanosight technology was used to evaluate microparticle size distribution. Particulate matter induces a dose- and time- dependent, rapid (1h) increase in microparticle generation in both cells. These microparticles express functional tissue factor. Particulate matter increases intracellular calcium concentration and phospholipase C inhibition reduces microparticle generation. Nanosight analysis confirmed that upon exposure to particulate matter both cells express particles with a size range consistent with the definition of microparticles (50-1000 nm). Exposure of mononuclear and endothelial cells to particulate matter upregulates the generation of microparticles at least partially mediated by calcium mobilization. This observation might provide a further link between airborne pollution and cardiopulmonary morbidity.
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- 2016
4. New insights in enumeration methodologies of probiotic cells in finished products
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Chiara Foglia, Annachiara De Prisco, Angela Amoruso, Marco Pane, and Serena Allesina
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Microbiology (medical) ,Combined use ,Cell ,Colony Count, Microbial ,Context (language use) ,Biology ,Microbiology ,law.invention ,Flow cytometry ,03 medical and health sciences ,Probiotic ,Lactobacillus rhamnosus ,law ,Enumeration ,medicine ,Food science ,Molecular Biology ,030304 developmental biology ,0303 health sciences ,Microbial Viability ,medicine.diagnostic_test ,Lacticaseibacillus rhamnosus ,030306 microbiology ,Probiotics ,Replicate ,Flow Cytometry ,biology.organism_classification ,medicine.anatomical_structure - Abstract
The number of bacterial cells is currently recognized as the most important parameter for the efficacy and quality of finished probiotic or live biotherapeutic products (LBP). Cell enumeration is generally performed by culture-dependent methodologies like plate count (PC). These techniques are able to reveal the number of viable cells able to replicate and generate a colony. However, they are limited by their dependence on the combination of culture conditions (e.g. nutrients, temperature) selected for cell recovery. Additionally, they do not provide information on the heterogeneity of a bacterial culture, namely they do not detect the cells in a viable but not cultivable (VBNC) status. Flow-cytometry (FC) is a culture-independent methodology having the potential to enumerate selectively live and damaged or dead cells. FC relies on the use of specific probes for different cell targets (e.g. membrane, enzymes) to unveil information on the cell structure and physiological statuses within a bacterial population. In this context, we monitored three batches of freeze-dried Lactobacillus rhamnosus GG (ATCC 53103) during a 3 year of storage at different conditions of temperature and relative humidity, according to ICH guidelines, by means of PC and FC. The Arrhenius model was applied to assess the suitability of the model to predict the mortality of probiotic cells in finished products. The higher destruction rate (k) obtained by PC data compared to FC data suggests a faster reduction of cultivability compared to membrane integrity, probably representing a dynamic shift of the bacterial population into a VBNC/dormant status during storage time. Interestingly, this mechanistic approach works both for PC and FC methodologies increasing the chances to monitor biological phenomenon within a mathematical modelling. The combined use of PC and FC shed lights on the true bacterial potency within a closed system like a finished product and the complexity of its heterogeneity.
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- 2020
5. Neurokinin (NK)-1 receptor expression in monocytes from bipolar disorder patients: A pilot study
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Angela Amoruso, Carlo Ignazio Cattaneo, Claudio Bardelli, Luigia Grazia Fresu, Elena Manzetti, and Sandra Brunelleschi
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Adult ,Male ,Bipolar Disorder ,P50 ,Receptor expression ,Blotting, Western ,Pilot Projects ,Substance P ,Disease ,Monocytes ,Tobacco smoke ,chemistry.chemical_compound ,Western blot ,medicine ,Humans ,Bipolar disorder ,Receptor ,medicine.diagnostic_test ,business.industry ,NF-kappa B ,Middle Aged ,Receptors, Neurokinin-1 ,medicine.disease ,Psychiatry and Mental health ,Clinical Psychology ,Gene Expression Regulation ,chemistry ,Immunology ,Female ,Sodium-Potassium-Exchanging ATPase ,business ,Signal Transduction - Abstract
Neurokinin 1 receptors (NK-1R) have been involved in several psychiatric disorders including major depression, but less is known for bipolar disorder (BD).We compared NK-1R expression and Substance P (SP) ability to induce NF-κB activation in monocytes from BD patients and healthy donors (HD), also looking for the effects of tobacco smoke. After informed written consent, 20 euthymic BD patients, either bipolar type 1 (BDI) or type 2 (BDII), and 14 age-matched healthy donors (HD) were enrolled. NK-1R expression in monocytes was evaluated by Western blot and expressed as the ratio between NK-1R and Na(+)/K(+)-ATPase protein expressions. NF-κB activation was assessed by measuring the nuclear content of the p50 subunit (ELISA kit).NK-1R expression was significantly reduced (P0.001) in monocytes from BD patients as compared to HD, with no major differences between BDI and BDII patients. Tobacco smoke enhanced NK-1R expression in HD, but not in BD patients. Un-stimulated monocytes from BD patients presented a constitutively higher (P0.05) content of nuclear p50 subunit as compared to HD. SP and an NK-1R agonist induced NF-κB activation, with a higher effect in HD: this effect was receptor-mediated as it was abrogated by an NK-1R antagonist.As a pilot study enrolling 20 BD patients, an obvious limitation is the sample size.Our results show the existence of a relevant alteration in NK-1R expression in BD patients and further suggest SP involvement in BD, so improving our understanding of the underlying mechanisms of this disease.
- Published
- 2015
6. Characterization of the anti-inflammatory properties of NCX 429, a dual-acting compound releasing nitric oxide and naproxen
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Jesmond Dalli, Mauro Perretti, Luigia Grazia Fresu, Sandra Brunelleschi, Daniela Miglietta, Donata Federici Canova, Angela Amoruso, and Claudio Bardelli
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Naproxen ,medicine.drug_class ,Interleukin-1beta ,Analgesic ,Drug Evaluation, Preclinical ,Inflammation ,Peritonitis ,Pharmacology ,Nitric Oxide ,Dinoprostone ,General Biochemistry, Genetics and Molecular Biology ,Anti-inflammatory ,Nitric oxide ,Mice ,chemistry.chemical_compound ,Phagocytosis ,In vivo ,medicine ,Animals ,Humans ,Nitric Oxide Donors ,General Pharmacology, Toxicology and Pharmaceutics ,Nitrates ,biology ,Anti-Inflammatory Agents, Non-Steroidal ,NF-kappa B ,General Medicine ,In vitro ,Disease Models, Animal ,Matrix Metalloproteinase 9 ,chemistry ,biology.protein ,Cyclooxygenase ,medicine.symptom ,medicine.drug - Abstract
Aims Cyclooxygenase (COX)-inhibiting nitric oxide donors (CINODs) are a new class of drugs that structurally combine a COX inhibitor with a nitric oxide (NO) donating moiety. This combination reduces potential toxicity of the non-steroidal anti-inflammatory drugs (NSAIDs) whilst maintaining the analgesic and anti-inflammatory effects. The present study was undertaken to investigate the anti-inflammatory effects of NCX 429, a naproxen-based CINOD, and to assess the additional properties of NO donation beyond those related to naproxen. Main methods We evaluated the in vitro effects of NCX 429 on oxy-radical production, phagocytosis, cytokine release, MMP-9, PPARγ expression and NF-κB activation in human monocytes/MDM and compared to naproxen. Moreover, we compared the in vivo efficacy of NCX 429 and naproxen in a murine model of peritonitis. Key findings In all the experiments performed in vitro, NCX 429 reduced the inflammatory responses with equal or higher efficacy compared to naproxen. Moreover, in in vivo experiments, NCX 429, at the lowest dose tested, was able to significantly inhibit cell influx in response to IL-1β administration although naproxen was found to be more potent than NCX 429 at reducing PGE 2 in inflammatory exudates. Significance These results demonstrate that both in vitro and in vivo – in a murine model of peritonitis – NCX 429 elicits significant anti-inflammatory activity, beyond the simple COX inhibition or pure NO release. Therefore, NO donation along with COX inhibition may represent a strategy for investigating inflammatory diseases in which pain and function are not fully resolved by analgesics/anti-inflammatory drugs.
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- 2015
7. Montelukast prevents microparticle-induced inflammatory and functional alterations in human bronchial smooth muscle cells
- Author
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Sandra Brunelleschi, Claudio Bardelli, Alessandro Celi, Maria Cristina Breschi, Angela Amoruso, Fabio Stefanelli, Stefano Fogli, and Tommaso Neri
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Cyclopropanes ,Cell signaling ,medicine.medical_treatment ,Myocytes, Smooth Muscle ,Active Transport, Cell Nucleus ,Heterologous ,Bronchi ,Acetates ,Sulfides ,Pharmacology ,Monocytes ,Cell Line ,Pathogenesis ,Cell-Derived Microparticles ,Receptors, Adrenergic, beta ,Gene expression ,medicine ,Humans ,Anti-Asthmatic Agents ,Autocrine signalling ,Cells, Cultured ,Montelukast ,business.industry ,Interleukin-8 ,NF-kappa B ,Cytokine ,Gene Expression Regulation ,Cyclooxygenase 2 ,Quinolines ,business ,Intracellular ,Signal Transduction ,medicine.drug - Abstract
Microparticles (MPs) are membrane fragments that may play a role in the pathogenesis of chronic respiratory diseases. We aimed to investigate whether human monocytes/macrophage-derived MPs could induce a pro-inflammatory phenotype in human bronchial smooth muscle cells (BSMC) and the effect of montelukast in this setting. Experimental methods included isolation of human monocytes/macrophages and generation of monocyte-derived MPs, RT-PCR analysis of gene expression, immunoenzymatic determination of pro-inflammatory factor release, bioluminescent assay of intracellular cAMP levels and electromobility shift assay analysis of NF-κB nuclear translocation. Stimulation of human BSMC with monocyte-derived MPs induced a pro-inflammatory switch in human BSMC by inducing gene expression (COX-2 and IL-8), protein release in the supernatant (PGE2 and IL-8), and heterologous β2-adrenoceptor desensitization. The latter effect was most likely related to autocrine PGE2 since pre-treatment with COX inhibitors restored the ability of salbutamol to induce cAMP synthesis in desensitized cells. Challenge with MPs induced nuclear translocation of NF-κB and selective NF-κB inhibition decreased MP-induced cytokine release in the supernatant. Montelukast treatment prevented IL-8 release and heterologous β2-adrenoceptor desensitization in human BSMC exposed to monocyte-derived MPs by blocking NF-κB nuclear translocation. These findings provide evidence on the role of human monocyte-derived MPs in the airway smooth muscle phenotype switch as a novel potential mechanism in the progression of chronic respiratory diseases and on the protective effects by montelukast in this setting.
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- 2013
8. Quantification of PPAR-γ protein in monocyte/macrophages from healthy smokers and non-smokers: A possible direct effect of nicotine
- Author
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Claudio Bardelli, L. Fresu, Sandra Brunelleschi, Gabriele Gunella, Valeria Ferrero, and Angela Amoruso
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Adult ,Male ,Nicotine ,medicine.medical_specialty ,alpha7 Nicotinic Acetylcholine Receptor ,medicine.medical_treatment ,Receptors, Nicotinic ,Gene Expression Regulation, Enzymologic ,Monocytes ,General Biochemistry, Genetics and Molecular Biology ,Ciglitazone ,Internal medicine ,medicine ,Humans ,Macrophage ,General Pharmacology, Toxicology and Pharmaceutics ,Interleukin 6 ,biology ,Interleukin-6 ,Tumor Necrosis Factor-alpha ,Macrophages ,Monocyte ,Smoking ,General Medicine ,Actins ,Up-Regulation ,PPAR gamma ,Endocrinology ,medicine.anatomical_structure ,Cytokine ,biology.protein ,Female ,Thiazolidinediones ,Cytokine secretion ,Tumor necrosis factor alpha ,medicine.drug - Abstract
Previous observations demonstrated that Peroxisome Proliferator-Activated Receptor-gamma (PPAR-gamma), a key regulator of adipocyte differentiation, is expressed in a large variety of cells, including cells of the monocyte/macrophage lineage. This study was aimed to quantify both the constitutive and ligand-induced PPAR-gamma expression in monocytes and monocyte-derived macrophages (MDM) isolated from healthy smokers and non-smokers, and to evaluate the possible direct effect of nicotine. PPAR-gamma protein was detected by Western blot and quantification was performed by calculating the ratio between PPAR-gamma and beta-actin protein expression. Cytokine release was measured with enzyme-linked immunoassay kits. Constitutive PPAR-gamma protein was detected in human monocytes and its expression was up-regulated along with differentiation to MDM. The endogenous ligand 15-deoxy-delta(12,14)-prostaglandin J(2) and the synthetic agonist ciglitazone enhanced PPAR-gamma expression, the former being effective also at low micromolar concentrations. Both agonists significantly inhibited the basal secretion of pro-inflammatory cytokines (e.g., TNF-alpha, IL-6), ciglitazone being more potent. Monocytes and MDM from healthy smokers presented a significantly enhanced (4-fold and 2.5-fold, respectively) constitutive PPAR-gamma expression, as compared to those from healthy non-smokers. However, ligand-induced PPAR-gamma expression and inhibition of cytokine secretion were similar in healthy smokers and non-smokers. Nicotine dose-dependently enhanced PPAR-gamma expression with a maximum at 10 muM, and inhibited release of pro-inflammatory cytokines; these effects were reversed by alpha-bungarotoxin. Nicotine and PPAR-gamma agonists did not exert synergistic effects. In conclusion, monocytes and MDM from healthy smokers present a constitutively enhanced PPAR-gamma expression; this effect is reproduced, to some extent, by nicotine in vitro.
- Published
- 2007
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