307 results on '"Cuda G"'
Search Results
52. Skeletal muscle expression and abnormal function of beta-myosin in hypertrophic cardiomyopathy.
- Author
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Cuda, G, primary, Fananapazir, L, additional, Zhu, W S, additional, Sellers, J R, additional, and Epstein, N D, additional
- Published
- 1993
- Full Text
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53. Modeling and designing a proteomics application on PROTEUS.
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Cannataro, M., Cuda, G., and Veltri, P.
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BIOINFORMATICS ,BREAST cancer ,PROTEOMICS ,MASS spectrometry ,COMPUTERS in biology ,GENETIC mutation ,ALGORITHMS ,COMPARATIVE studies ,DATABASES ,INFORMATION retrieval ,INTERNATIONAL relations ,INTERNET ,MANAGEMENT information systems ,RESEARCH methodology ,MEDICAL cooperation ,MEDICAL informatics ,PROBLEM solving ,RESEARCH ,SYSTEM integration ,EVALUATION research ,HUMAN services programs - Abstract
Objectives: Biomedical applications, such as analysis and management of mass spectrometry proteomics experiments, involve heterogeneous platforms and knowledge, massive data sets, and complex algorithms. Main requirements of such applications are semantic modeling of the experiments and data analysis, as well as high performance computational platforms. In this paper we propose a software platform allowing to model and execute biomedical applications on the Grid.Methods: Computational Grids offer the required computational power, whereas ontologies and workflow help to face the heterogeneity of biomedical applications. In this paper we propose the use of domain ontologies and workflow techniques for modeling biomedical applications, whereas Grid middleware is responsible for high performance execution. As a case study, the modeling of a proteomics experiment is discussed.Results: The main result is the design and first use of PROTEUS, a Grid-based problem-solving environment for biomedical and bioinformatics applications.Conclusion: To manage the complexity of biomedical experiments, ontologies help to model applications and to identify appropriate data and algorithms, workflow techniques allow to combine the elements of such applications in a systematic way. Finally, translation of workflow into execution plans allows the exploitation of the computational power of Grids. Along this direction, in this paper we present PROTEUS discussing a real case study in the proteomics domain. [ABSTRACT FROM AUTHOR]- Published
- 2005
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54. Malignant ventricular arrhythmia in the wolff-parkinson-white syndrome during amiodarone treatment.
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Perticone, F., Cuda, G., Spadea, F., Pintaudi, C., and Tropea, R.
- Published
- 1987
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55. Transcriptional regulation of the mismatch repair gene hMLH1
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Quaresima, B., Faniello, M. C., Baudi, F., Cuda, G., Grandinetti, C., Tassone, P., Costanzo, F., and Venuta, S.
- Published
- 2001
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56. Effects of glucocorticoids on activation of c-jun N-terminal, extracellular signal-regulated, and p38 MAP kinases in human pulmonary endothelial cells
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Pelaia, G., Cuda, G., Vatrella, A., Grembiale, R. D., Sarro, G. De, Maselli, R., Costanzo, F. S., Avvedimento, V. E., Rotiroti, D., and Marsico, S. A.
- Published
- 2001
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57. Changes in myocardial cytoskeletal intermediate filaments and myocyte contractile dysfunction in dilated cardiomyopathy: an in vivo study in humans
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Vivo, F. de, Somma, S. Di, Marotta, M., Salvatore, G., Cudemo, G., Benedetto, M.P. Di, Caputo, G., Divitiis, O. de, Cuda, G., and Ciaramella, F.
- Abstract
AimTo investigate in vivo the intermediate cytoskeletal filaments desmin and vimentin in myocardial tissues from patients with dilated cardiomyopathy, and to determine whether alterations in these proteins are associated with impaired contractility.MethodsEndomyocardial biopsies were performed in 12 patients with dilated cardiomyopathy and in 12 controls (six women with breast cancer before anthracycline chemotherapy and six male donors for heart transplantation). Biopsy specimens were analysed by light microscopy and immunochemistry (desmin, vimentin). Myocyte contractile protein function was evaluated by the actin-myosin in vitro motility assay. Left ventricular ejection fraction was assessed by echocardiography and radionuclide ventriculography.ResultsPatients with dilated cardiomyopathy had a greater cardiomyocyte diameter than controls (p < 0.01). The increase in cell size was associated with a reduction in contractile function, as assessed by actin-myosin motility (r = -0.643; p < 0.01). Quantitative immunochemistry showed increased desmin and vimentin contents (p < 0.01), and the desmin distribution was disturbed in cardiomyopathy. There was a linear relation between desmin distribution and actin-myosin sliding in vitro (r = 0.853; p < 0.01) and an inverse correlation between desmin content and ejection fraction (r = -0.773; p < 0.02). Negative correlations were also found between myocardial vimentin content and the actin-myosin sliding rate (r = -0.74; p < 0.02) and left ventricular ejection fraction (r = -0.68; p < 0.01).ConclusionsCompared with normal individuals, the myocardial tissue of patients with dilated cardiomyopathy shows alterations of cytoskeletal intermediate filament distribution and content associated with reduced myocyte contraction.
- Published
- 2000
58. Detection of microsatellite instability and loss of heterozygosity in serum DNA of small and non-small cell lung cancer patients: a tool for early diagnosis?
- Author
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Cuda, G., Gallelli, A., Nistico, A., Tassone, P., Barbieri, V., Tagliaferri, P. S., Costanzo, F. S., Tranfa, C. M., and Venuta, S.
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- 2000
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59. Influence of the cardiac myosin hinge region on contractile activity.
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Margossian, S S, Krueger, J W, Sellers, J R, Cuda, G, Caulfield, J B, Norton, P, and Slayter, H S
- Abstract
The participation of cardiac myosin hinge in contractility was investigated by in vitro motility and ATPase assays and by measurements of sarcomere shortening. The effect on contractile activity was analyzed using an antibody directed against a 20-amino acid peptide within the hinge region of myosin. This antibody bound specifically at the hinge at a distance of 55 nm from the S1/S2 junction, was specific to human, dog, and rat cardiac myosins, did not crossreact with gizzard or skeletal myosin, and had no effect on ATPase activity of purified S1 and myofibrils. However, it completely suppressed the movement of actin filaments in in vitro motility assays and reduced active shortening of sarcomeres of skinned cardiac myocytes by half. Suppression of motion by the anti-hinge antibody may reflect a mechanical constraint imposed by the antibody upon the mobility of the S2 region of myosin. The results suggest that the steps in the mechanochemical energy transduction can be separately influenced through S2.
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- 1991
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60. In situ microfluidic biofunctionalisation to form multivalent interactions and investigate cell rolling and phenotype modification
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Perozziello, G., Simone, G., La Rocca, R., Pardeo, F., Candeloro, P., Natalia MALARA, Liberale, C., Angelis, F., Cuda, G., Carbone, E., and Di Fabrizio, E.
61. Effects of endothelin-1 on MAPK activation, IL-6 production and cell proliferation in normal human lung fibroblasts
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Gallelli, L., Pelaia, G., Cuda, G., Alessandro Vatrella, Fratto, D., Gioffrè, V., D’agostino, B., Mastruzzo, C., Sofia, M., Maniscalco, M., Crimi, N., Maselli, R., Rossi, F., Costanzo, F. S., Marsico, S. A., and Vancheri, C.
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IL-6 ,lung fibroblasts ,Endothelin ,MAPK
62. PACLITAXEL RESISTANCE OF BRCA1 MUTATED HCC1937 BREAST CANCER CELLS CORRELATES WITH CHANGES IN THE WHOLE CELL PROTEOME
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Blotta, S., Casadonte, R., Camillo Palmieri, Quaresima, B., Pietragalla, A., Lotti, Lv, Terracciano, R., Cuda, G., Tagliaferri, P., Tassone, P., Costantini, Rm, and Venuta, S.
63. Proteomic profiling of inherited breast cancer: Identification of molecular targets for early detection, prognosis and treatment, and related bioinformatics tools
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Cuda, G., Cannataro, M., Quaresima, B., Baudi, F., Casadonte, R., Faniello, M. C., Tagliaferri, P., Pierangelo VELTRI, Costanzo, F., and Venuta, S.
64. Effects of TGF-beta and budesonide on MAPK phosphorylation, IL-6/IL-11 secretion, and cell proliferation in human lung fibroblasts
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Pelaia, G., Gallelli, L., D’agostino, B., Alessandro Vatrella, Cuda, G., Fratto, D., Renda, T., Galderisi, U., Piegari, E., Crimi, N., Rossi, F., Caputi, M., Costanzo, F. S., Vancheri, C., Maselli, R., and Marsico, S. A.
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IL-6 ,tgf-beta ,budesonide ,mapk ,IL-11 ,lung fibroblasts
65. An interactive tool for the management and visualization of mass-spectrometry proteomics data
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Mario Cannataro, Cuda, G., Gaspari, M., and Veltri, P.
66. Microfluidic polyimide chips fabricated by lamination processes for X-ray scattering applications
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Perozziello, G., Catalano, R., Simone, G., Candeloro, P., Malara, N., Santoriello, S., La Rocca, R., Angelis, F., Angelo Accardo, Burghammer, M., Di Cola, E., Cuda, G., Riekel, C., and Di Fabrizio, E.
67. Inhibition of myocardial crossbridge cycling by hypoxic endothelial cells: A potential mechanism for matching oxygen supply and demand?
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Ajay Shah, Mebazaa, A., Yang, Z. -K, Cuda, G., Lankford, E. B., Pepper, C. B., Sollott, S. J., Sellers, J. R., Robotham, J. L., and Lakatta, E. G.
68. Effects of MAPK inhibitors on H2O2-induced IL-8 production and cell death in primary cultures of human bronchial epithelial cells
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Pelaia, G., Cuda, G., Alessandro Vatrella, Gallelli, L., Fratto, D., Gioffrè, V., Matera, M. G., D’agostino, B., Caputi, M., Cazzola, M., Maselli, R., Rossi, F., Costanzo, F. S., and Marsico, S. A.
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IL-8 ,H2O2 ,airway epithelial cell ,MAPK
69. The EIPeptiDi tool: enhancing peptide discovery in ICAT-based LC MS/MS experiments
- Author
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Tradigo Giuseppe, Greco Sergio, Gaspari Marco, Cuda Giovanni, Cannataro Mario, and Veltri Pierangelo
- Subjects
Computer applications to medicine. Medical informatics ,R858-859.7 ,Biology (General) ,QH301-705.5 - Abstract
Abstract Background Isotope-coded affinity tags (ICAT) is a method for quantitative proteomics based on differential isotopic labeling, sample digestion and mass spectrometry (MS). The method allows the identification and relative quantification of proteins present in two samples and consists of the following phases. First, cysteine residues are either labeled using the ICAT Light or ICAT Heavy reagent (having identical chemical properties but different masses). Then, after whole sample digestion, the labeled peptides are captured selectively using the biotin tag contained in both ICAT reagents. Finally, the simplified peptide mixture is analyzed by nanoscale liquid chromatography-tandem mass spectrometry (LC-MS/MS). Nevertheless, the ICAT LC-MS/MS method still suffers from insufficient sample-to-sample reproducibility on peptide identification. In particular, the number and the type of peptides identified in different experiments can vary considerably and, thus, the statistical (comparative) analysis of sample sets is very challenging. Low information overlap at the peptide and, consequently, at the protein level, is very detrimental in situations where the number of samples to be analyzed is high. Results We designed a method for improving the data processing and peptide identification in sample sets subjected to ICAT labeling and LC-MS/MS analysis, based on cross validating MS/MS results. Such a method has been implemented in a tool, called EIPeptiDi, which boosts the ICAT data analysis software improving peptide identification throughout the input data set. Heavy/Light (H/L) pairs quantified but not identified by the MS/MS routine, are assigned to peptide sequences identified in other samples, by using similarity criteria based on chromatographic retention time and Heavy/Light mass attributes. EIPeptiDi significantly improves the number of identified peptides per sample, proving that the proposed method has a considerable impact on the protein identification process and, consequently, on the amount of potentially critical information in clinical studies. The EIPeptiDi tool is available at http://bioingegneria.unicz.it/~veltri/projects/eipeptidi/ with a demo data set. Conclusion EIPeptiDi significantly increases the number of peptides identified and quantified in analyzed samples, thus reducing the number of unassigned H/L pairs and allowing a better comparative analysis of sample data sets.
- Published
- 2007
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70. B006: Molecular and survival effects of amlodipine on endothelial cells stressed with h2o2.
- Author
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Perticone, F., Ceravolo, R., Candigliota, M., Mongiardo, A., and Cuda, G.
- Published
- 2000
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71. B006: Molecular and survival effects of amlodipine on endothelial cells stressed with h2o2
- Author
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Perticone, F., Ceravolo, R., Candigliota, M., Mongiardo, A., and Cuda, G.
- Abstract
Cellular stress elicits biochemical responses that either enhance cells survival or lead to cells death or apoptosis. The Ca2+ involvement in apoptosis was demonstrated by several experimental studies using calcium channel blockers (CCBs). Thus, Ca2+ increasing could be an important trigger mechanism for apoptosis. However, the CCBs effects on apoptosis, cells proliferation, cells death induced by oxidative stress remains still unclear. The aim of our study was to assess the effects of amlodipine (AML) (a dydropyridine CCBs) on humbelical vein endothelial cells (HUVEc) stressed with H
2 O2 (2 mM). We tested the cells death by triphan bleu, and apoptosis by annexin and caspases. The oxidative stress (2 hrs of H2 O2 administration) on HUVEc induced a 22% of cells death (Fig. A ) and a 20% of apoptosis (Fig. C ). On the other hand, AML overnight pretreatment induced a significant reduction of cells death (Fig. B ) and apoptosis (Fig. C ). The free radical formation (dyclo-roflurescein) in unstressed HUVEc was also significantly reduced by AML treatment (p < 0.0001, by ANOVA). In conclusion, the AML pretreatment reduces cells death and apoptosis, probably, by its antioxidant effects.- Published
- 2000
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72. Effects of TGF-beta and glucocorticoids on map kinase phosphorylation, IL-6/IL-11 secretion and cell proliferation in primary cultures of human lung fibroblasts
- Author
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Francesco Rossi, Alessandro Vatrella, Teresa Renda, Serafino A. Marsico, Rosario Maselli, Nunzio Crimi, Francesco Costanzo, Elena Piegari, Mario Caputi, Umberto Galderisi, D. Fratto, Luca Gallelli, Carlo Vancheri, Girolamo Pelaia, Giovanni Cuda, Bruno D'Agostino, Pelaia, G, Gallelli, L, D'Agostino, Bruno, Vatrella, A, Cuda, G, Fratto, D, Renda, T, Galderisi, Umberto, Piegari, Elena, Crimi, N, Rossi, Francesco, Caputi, M, Costanzo, F, Vancheri, C, Maselli, R, Marsico, Sa, Pelaia, G., Gallelli, L., D'Agostino, B., Vatrella, Alessandro, Cuda, G., Fratto, D., Renda, T., Galderisi, U., Piegari, E., Crimi, N., Rossi, F., Caputi, M., Costanzo, F. S., Vancheri, C., Maselli, R., and Marsico, S. A.
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MAPK/ERK pathway ,mitogen-activated protein kinase ,budesonide ,glucocorticoid ,interleukin 11 ,interleukin 6 ,transforming growth factor beta ,mapk ,Physiology ,medicine.medical_treatment ,Clinical Biochemistry ,TGF-beta ,Enzyme Inhibitors ,Phosphorylation ,Lung ,Cells, Cultured ,interstitial lung disease ,Mitogen-Activated Protein Kinase 1 ,interleukin ,Interleukin-11 ,Up-Regulation ,Human Lung Fibroblast ,Map Kinase ,medicine.medical_specialty ,Cell Survival ,MAP Kinase Signaling System ,Biology ,Transforming Growth Factor beta1 ,Internal medicine ,TGF beta signaling pathway ,medicine ,Humans ,Viability assay ,Protein kinase A ,Glucocorticoids ,Cell Proliferation ,Cell growth ,Interleukin-6 ,Growth factor ,Interleukins ,Cell Biology ,Transforming growth factor beta ,Fibroblasts ,Molecular biology ,TGF-b ,Enzyme Activation ,Endocrinology ,biology.protein - Abstract
Transforming growth factor-beta1 (TGF-beta1) is crucially involved in the fibrotic events characterizing interstitial lung diseases (ILDs), as well as in the airway remodeling process typical of asthma. Within such a context, the aim of our study was to investigate, in primary cultures of normal and fibrotic human lung fibroblasts (HLFs), the effects of TGF-beta1 on mitogen-activated protein kinase (MAPK) phosphorylation, cell proliferation, and production of interleukins 6 (IL-6) and 11 (IL-11), in the presence or absence of a pretreatment with budesonide (BUD). MAPK phosphorylation was detected by Western blotting, cell viability and proliferation were evaluated using Trypan blue staining and [(3)H]-thymidine incorporation assay, respectively, and the release of IL-6 and IL-11 into cell culture supernatants was assessed by ELISA. TGF-beta1 (10 ng/ml) significantly stimulated MAPK phosphorylation (P < 0.01), and also enhanced cell proliferation as well as the secretion of both IL-6 and IL-11, which reached the highest increases at the 72nd h of cell exposure to this growth factor. All such effects were prevented by BUD (10(-8) M) and, with the exception of IL-6 release, also by a mixture of MAPK inhibitors. Therefore, our findings suggest that the fibrotic action exerted by TGF-beta1 in the lung is mediated at least in part by MAPK activation and by an increased synthesis of the profibrogenic cytokines IL-6 and IL-11; all these effects appear to be prevented by corticosteroids via inhibition of MAPK phosphorylation.
- Published
- 2007
73. Moving beyond the Tip of the Iceberg: DJ-1 Implications in Cancer Metabolism
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Erika Olivo, Marina La Chimia, Jessica Ceramella, Alessia Catalano, Ferdinando Chiaradonna, Maria Stefania Sinicropi, Giovanni Cuda, Domenico Iacopetta, Domenica Scumaci, Olivo, E, Chimia, M, Ceramella, J, Catalano, A, Chiaradonna, F, Sinicropi, M, Cuda, G, Iacopetta, D, and Scumaci, D
- Subjects
PARK7 ,Oxidative Stress ,DJ-1 ,Neoplasms ,Protein Deglycase DJ-1 ,Humans ,cancer metabolism ,Neurodegenerative Diseases ,Parkinson Disease ,General Medicine ,Reactive Oxygen Species ,Oxidation-Reduction ,ferroptosi - Abstract
DJ-1, also called Parkinson’s protein 7 (PARK7), is ubiquitously expressed and plays multiple actions in different physiological and, especially, pathophysiological processes, as evidenced by its identification in neurodegenerative diseases and its high expression in different types of cancer. To date, the exact activity of DJ-1 in carcinogenesis has not been fully elucidated, however several recent studies disclosed its involvement in regulating fundamental pathways involved in cancer onset, development, and metastatization. At this purpose, we have dissected the role of DJ-1 in maintaining the transformed phenotype, survival, drug resistance, metastasis formation, and differentiation in cancer cells. Moreover, we have discussed the role of DJ-1 in controlling the redox status in cancer cells, along with the ability to attenuate reactive oxygen species (ROS)-dependent cell death, as well as to mediate ferropotosis. Finally, a mention to the development of therapeutic strategies targeting DJ-1 has been done. We have reported the most recent studies, aiming to shed light on the role played by DJ-1 in different cancer aspects and create the foundation for moving beyond the tip of the iceberg.
- Published
- 2022
74. Statins Stimulate New Myocyte Formation After Myocardial Infarction by Activating Growth and Differentiation of the Endogenous Cardiac Stem Cells
- Author
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Mariangela Scalise, Bernardo Nadal-Ginard, Fabiola Marino, Liberato Berrino, Daniele Torella, Teresa Mancuso, Elvira Immacolata Parrotta, Francesco Rossi, Eleonora Cianflone, Donato Cappetta, Giovanni Cuda, Konrad Urbanek, Alessandro Salatino, Michele Albanese, Antonella De Angelis, Jolanda Sabatino, Cianflone, E., Cappetta, D., Mancuso, T., Sabatino, J., Marino, F., Scalise, M., Albanese, M., Salatino, A., Parrotta, E. I., Cuda, G., De Angelis, A., Berrino, L., Rossi, F., Nadal-Ginard, B., Torella, D., and Urbanek, K.
- Subjects
0301 basic medicine ,Simvastatin ,Myocardial Infarction ,030204 cardiovascular system & hematology ,Cardiac stem cell ,lcsh:Chemistry ,Mice ,0302 clinical medicine ,3-hydroxy-3-methylglutaryl coenzyme A ,Myocardial infarction ,Phosphorylation ,Rosuvastatin Calcium ,lcsh:QH301-705.5 ,Cells, Cultured ,Spectroscopy ,Pravastatin ,Cultured ,Akt ,Cardiac stem cells ,Myocardial regeneration ,Statins ,Animals ,Cell Differentiation ,Cell Proliferation ,Disease Models, Animal ,Female ,Hydroxymethylglutaryl-CoA Reductase Inhibitors ,Muscle Cells ,Myocardium ,Proto-Oncogene Proteins c-akt ,Rats ,Stem Cells ,cardiac stem cells ,General Medicine ,Computer Science Applications ,myocardial regeneration ,Stem cell ,medicine.drug ,Cells ,Article ,Catalysis ,Inorganic Chemistry ,03 medical and health sciences ,medicine ,Rosuvastatin ,cardiovascular diseases ,Physical and Theoretical Chemistry ,Progenitor cell ,Molecular Biology ,Protein kinase B ,Animal ,business.industry ,Cell growth ,Organic Chemistry ,nutritional and metabolic diseases ,medicine.disease ,030104 developmental biology ,lcsh:Biology (General) ,lcsh:QD1-999 ,Disease Models ,Cancer research ,business - Abstract
The 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase inhibitors (statins) exert pleiotropic effects on cardiac cell biology which are not yet fully understood. Here we tested whether statin treatment affects resident endogenous cardiac stem/progenitor cell (CSC) activation in vitro and in vivo after myocardial infarction (MI). Statins (Rosuvastatin, Simvastatin and Pravastatin) significantly increased CSC expansion in vitro as measured by both BrdU incorporation and cell growth curve. Additionally, statins increased CSC clonal expansion and cardiosphere formation. The effects of statins on CSC growth and differentiation depended on Akt phosphorylation. Twenty-eight days after myocardial infarction by permanent coronary ligation in rats, the number of endogenous CSCs in the infarct border zone was significantly increased by Rosuvastatin-treatment as compared to untreated controls. Additionally, commitment of the activated CSCs into the myogenic lineage (c-kitpos/Gata4pos CSCs) was increased by Rosuvastatin administration. Accordingly, Rosuvastatin fostered new cardiomyocyte formation after MI. Finally, Rosuvastatin treatment reversed the cardiomyogenic defects of CSCs in c-kit haploinsufficient mice, increasing new cardiomyocyte formation by endogenous CSCs in these mice after myocardial infarction. In summary, statins, by sustaining Akt activation, foster CSC growth and differentiation in vitro and in vivo. The activation and differentiation of the endogenous CSC pool and consequent new myocyte formation by statins improve myocardial remodeling after coronary occlusion in rodents. Similar effects might contribute to the beneficial effects of statins on human cardiovascular diseases.
- Published
- 2020
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75. A passive microfluidic device for chemotaxis studies
- Author
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Maria Antonia D'Attimo, Ennio Carbone, Costanza Maria Cristiani, Enzo Di Fabrizio, Gerardo Perozziello, Ulrich Krühne, Giovanni Cuda, Francesco Guzzi, Elvira Immacolata Parrotta, Patrizio Candeloro, Elisabetta Dattola, Maria Laura Coluccio, E. Lamanna, Coluccio, M. L., D'Attimo, M. A., Cristiani, C. M., Candeloro, P., Parrotta, E., Dattola, E., Guzzi, F., Cuda, G., Lamanna, E., Carbone, E., Kruhne, U., Di Fabrizio, E., and Perozziello, G.
- Subjects
Materials science ,Microscope ,Gravity force ,Diffusion ,lcsh:Mechanical engineering and machinery ,Microfluidics ,02 engineering and technology ,Mini incubator ,01 natural sciences ,Passive microfluidic device ,Article ,law.invention ,law ,lcsh:TJ1-1570 ,Electrical and Electronic Engineering ,chemotaxis ,business.industry ,Mechanical Engineering ,Chemotaxis ,010401 analytical chemistry ,Incubator ,Chemotaxi ,021001 nanoscience & nanotechnology ,0104 chemical sciences ,Volumetric flow rate ,Control and Systems Engineering ,Optoelectronics ,0210 nano-technology ,business ,Concentration gradient - Abstract
This work presents a disposable passive microfluidic system, allowing chemotaxis studies, through the generation of a concentration gradient. The device can handle liquid flows without an external supply of pressure or electric gradients, but simply using gravity force. It is able to ensure flow rates of 10 µ, L/h decreasing linearly with 2.5% in 24 h. The device is made of poly(methylmethacrylate) (PMMA), a biocompatible material, and it is fabricated by micro-milling and solvent assisted bonding. It is assembled into a mini incubator, designed properly for cell biology studies in passive microfluidic devices, which provides control of temperature and humidity levels, a contamination-free environment for cells with air and 5% of CO2. Furthermore, the mini incubator can be mounted on standard inverted optical microscopes. By using our microfluidic device integrated into the mini incubator, we are able to evaluate and follow in real-time the migration of any cell line to a chemotactic agent. The device is validated by showing cell migration at a rate of 0.36 µ, m/min, comparable with the rates present in scientific literature.
- Published
- 2019
76. H ferritin silencing induces protein misfolding in K562 cells: A Raman analysis
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Fabiana Zolea, Anna Di Vito, Francesco Costanzo, Giovanni Cuda, Francesca Trecroci, Anna Cozzi, Nadia Lobello, Flavia Biamonte, Enzo Di Fabrizio, Maddalena Di Sanzo, Barbara Quaresima, Sonia Levi, Patrizio Candeloro, Zolea, F, Biamonte, F, Candeloro, P, Di Sanzo, M, Cozzi, A, Di Vito, A, Quaresima, B, Lobello, N, Trecroci, F, Di Fabrizio, E, Levi, SONIA MARIA ROSA, Cuda, G, and Costanzo, F.
- Subjects
Protein Folding ,DNA damage ,Protein subunit ,Fluorescent Antibody Technique ,Spectrum Analysis, Raman ,medicine.disease_cause ,Biochemistry ,Physiology (medical) ,medicine ,Homeostasis ,Humans ,Gene silencing ,chemistry.chemical_classification ,Reactive oxygen species ,biology ,Reverse Transcriptase Polymerase Chain Reaction ,Cell biology ,Ferritin ,Oxidative Stress ,chemistry ,Gene Knockdown Techniques ,Apoferritins ,biology.protein ,Protein folding ,K562 Cells ,Reactive Oxygen Species ,Oxidation-Reduction ,Intracellular ,Oxidative stress - Abstract
The redox state of the cell is involved in the regulation of many physiological functions as well as in the pathogenesis of several diseases, and is strictly dependent on the amount of iron in its catalytically active state. Alterations of iron homeostasis determine increased steady-state concentrations of Reactive Oxygen Species (ROS) that cause lipid peroxidation, DNA damage and altered protein folding. Ferritin keeps the intracellular iron in a non-toxic and readily available form and consequently plays a central role in iron and redox homeostasis. The protein is composed by 24 subunits of the H- and L-type, coded by two different genes, with structural and functional differences. The aim of this study was to shed light on the role of the single H ferritin subunit (FHC) in keeping the native correct protein three-dimensional structure. To this, we performed Raman spectroscopy on protein extracts from K562 cells subjected to FHC silencing. The results show a significant increase in the percentage of disordered structures content at a level comparable to that induced by H2O2 treatment in control cells. ROS inhibitor and iron chelator were able to revert protein misfolding. This integrated approach, involving Raman spectroscopy and targeted-gene silencing, indicates that an imbalance of the heavy-to-light chain ratio in the ferritin composition is able to induce severe but still reversible modifications in protein folding and uncovers new potential pathogenetic mechanisms associated to intracellular iron perturbation.
- Published
- 2015
77. Superhydrophobic lab-on-chip measures secretome protonation state and provides a personalized risk assessment of sporadic tumour
- Author
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S. Bonacci, Ivan Presta, Rosario Sacco, Elisabetta Ferraro, Nicola Coppedè, M. Greco, Natalia Malara, Ugo Bottoni, Domenica Scumaci, Giuseppe Donato, Gianni Cuda, Roksana Majewska, Volpentesta G, Francesco Gentile, A. Donato, Valentina Trunzo, Giusy Guzzi, Nadia Innaro, Domenico Augusto Francesco Maisano, Valentina Onesto, A. Castellini, C. K. Pirrone, P. Candeloro, Chiara Mignogna, Gerardo Perozziello, Francesco Amato, F. Casale, Maria Laura Coluccio, F. Givigliano, Lorenzo Ferrara, C. Voci, M. Renne, E. Di Fabrizio, Vincenzo Mollace, Marco Giannetto, Giuseppe Sena, Angelo Lavano, Elisabetta Scali, Maria Careri, Malara, N., Gentile, F., Coppedè, N., Coluccio, M. L., Candeloro, P., Perozziello, G., Ferrara, L., Giannetto, M., Careri, M., Castellini, A., Mignogna, C., Presta, I., Pirrone, C. K., Maisano, D., Donato, A., Donato, G., Greco, M., Scumaci, D., Cuda, G., Casale, F., Ferraro, E., Bonacci, S., Trunzo, V., Mollace, V., Onesto, V., Majewska, R., Amato, F., Renne, M., Innaro, N., Sena, G., Sacco, R., Givigliano, F., Voci, C., Volpentesta, G., Guzzi, G., Lavano, A., Scali, E., Bottoni, U., and Di Fabrizio, E.
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0301 basic medicine ,Oncology ,Cancer Research ,medicine.medical_specialty ,tumor early detection ,lab-on-a-chip ,business.industry ,lcsh:Neoplasms. Tumors. Oncology. Including cancer and carcinogens ,Cancer Early Detection ,lcsh:RC254-282 ,Article ,03 medical and health sciences ,030104 developmental biology ,0302 clinical medicine ,Text mining ,030220 oncology & carcinogenesis ,Internal medicine ,oect ,medicine ,False positive paradox ,Cancer risk ,business ,Risk assessment ,Sporadic cancer - Abstract
Secretome of primary cultures is an accessible source of biological markers compared to more complex and less decipherable mixtures such as serum or plasma. The protonation state (PS) of secretome reflects the metabolism of cells and can be used for cancer early detection. Here, we demonstrate a superhydrophobic organic electrochemical device that measures PS in a drop of secretome derived from liquid biopsies. Using data from the sensor and principal component analysis (PCA), we developed algorithms able to efficiently discriminate tumour patients from non-tumour patients. We then validated the results using mass spectrometry and biochemical analysis of samples. For the 36 patients across three independent cohorts, the method identified tumour patients with high sensitivity and identification as high as 100% (no false positives) with declared subjects at-risk, for sporadic cancer onset, by intermediate values of PS. This assay could impact on cancer risk management, individual’s diagnosis and/or help clarify risk in healthy populations., Diagnostics: Proton state of secreted proteins in blood helps identify cancer A blood test that measures whether molecules secreted by cells contain titratable proton atoms can accurately discriminate between patients who have cancer and those who don’t. Titratable species may in turn influence the protonation state of a solution, i.e. the number of protons added to and the net charge of that solution. A team led by Natalia Malara from University Magna Graecia in Catanzaro, Italy and Enzo Di Fabrizio from the King Abdullah University of Science and Technology in Thuwal, Saudi Arabia, Francesco Gentile from the University Federico II in Naples, Italy, and Nicola Coppedè from the Institute of Materials for Electronics and Magnetism in Parma, Italy, created an eletrochemical device that can detect faulty metabolism by quantifying the proportion of secreted proteins with and without extra protons—an indicator of abnormal cell division, proliferation and invasion. The researchers tested the device on blood samples from patients with solid tumors and healthy controls. The method identified cancer patients with a high degree of accuracy. If the findings are confirmed in larger trials, the test could help with the screening, diagnosis and management of cancer.
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- 2018
78. Proteomic analysis of protein purified derivative of Mycobacterium bovis
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Marco Gaspari, Franco Roperto, Valeria Russo, Roberta De Luca, Dora Maria Ceccarelli, Giovanni Cuda, Monica Cagiola, Sante Roperto, Mariaconcetta Varano, Roperto, Sante, Varano, M, Russo, Valeria, Luca', Roberta, Cagiola, M, Gaspari, M, Ceccarelli, DORA MARIA, Cuda, G, and Roperto, FRANCO PEPPINO
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Purify Protein Derivative ,0301 basic medicine ,Proteomics ,Proteomic Profile ,Protein digestion ,T-Lymphocytes ,lcsh:Medicine ,Tuberculin ,General Biochemistry, Genetics and Molecular Biology ,Microbiology ,03 medical and health sciences ,Bacterial Proteins ,Tandem Mass Spectrometry ,Tuberculosis ,Medicine ,Nanotechnology ,Tuberculin Skin Test ,Medicine(all) ,Mycobacterium bovis ,Antigens, Bacterial ,biology ,Molecular mass ,Staining and Labeling ,Biochemistry, Genetics and Molecular Biology(all) ,business.industry ,Research ,lcsh:R ,General Medicine ,biology.organism_classification ,030104 developmental biology ,Mycobacterium tuberculosis complex ,Proteome ,Tuberculosis Complex ,business ,Chromatography, Liquid - Abstract
Background Tuberculin skin test based on in vivo intradermal inoculation of purified protein derivative from Mycobacterium bovis (bPPD) is the diagnostic test for the control and surveillance of bovine tuberculosis (bTB). Methods Proteomic analysis was performed on different bPPD preparations from M. bovis, strain AN5. Proteins were precipitated from bPPD solutions by TCA precipitation. The proteome of bPPD preparations was investigated by bottom-up proteomics, which consisted in protein digestion and nano-LC–MS/MS analysis. Mass spectrometry analysis was performed on a Q-exactive hybrid quadrupole-Orbitrap mass spectrometer coupled online to an Easy nano-LC1000 system. Results Three hundred and fifty-six proteins were identified and quantified by at least 2 peptides (99% confidence per peptide). One hundred and ninety-eight proteins, which had not been previously described, were detected; furthermore, the proteomic profile shared 80 proteins with previous proteomes from bPPDs from the United Kingdom and Brazil and 139 protein components from bPPD from Korea. Locus name of M. bovis (Mb) with orthologs from M. tuberculosis H37Rv, comparative gene and protein length, molecular mass, functional categories, gene name and function of each protein were reported. Ninety-two T cell mycobacterial antigens responsible for delayed-type hypersensitivity were detected, fifty-two of which were not previously reported in any bPPD proteome. Data are available via ProteomeXchange with identifier PXD005920. Conclusions This study represents the highest proteome coverage of bPPD preparations to date. Since proteins perform cellular functions essential to health and/or disease, obtaining knowledge of their presence and variance is of great importance in understanding disease states and for advancing translational studies. Therefore, to better understand Mycobacterium tuberculosis complex biology during infection, survival, and persistence, the reproducible evaluation of the proteins that catalyze and control these processes is critically important. More active and more specific tuberculins would be desirable. Indeed, many antigens contained within bPPD are currently responsible for the cross-reactivity resulting in false-positive results as they are shared between non-tuberculous and tuberculous mycobacteria. Electronic supplementary material The online version of this article (doi:10.1186/s12967-017-1172-1) contains supplementary material, which is available to authorized users.
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- 2017
79. Microfluidics & nanotechnology: towards fully integrated analytical devices for the detection of cancer biomarkers
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Giovanni Cuda, E. Di Fabrizio, Patrizio Candeloro, Francesca Pardeo, Rossella Catalano, Horacio D. Espinosa, Andrea Adamo, Maria Laura Coluccio, Annalisa Nicastri, Elvira Immacolata Parrotta, Angela Mena Perri, Francesco Gentile, Gerardo Perozziello, Perozziello, G., Candeloro, P., Gentile, Francesco, Nicastri, A., Perri, A., Coluccio, M. L., Adamo, A., Pardeo, F., Catalano, R., Parrotta, E., Espinosa, H. D., Cuda, G., and Di Fabrizio, E.
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Materials science ,Resolution (mass spectrometry) ,General Chemical Engineering ,Chemistry (all) ,Microfluidics ,Nanotechnology ,General Chemistry ,symbols.namesake ,Small peptide ,symbols ,Coming out ,Chemical Engineering (all) ,Raman spectroscopy ,Biosensor ,Hydrodynamic flow ,Raman scattering - Abstract
In this paper, we describe an innovative modular microfluidic platform allowing filtering, concentration and analysis of peptides from a complex mixture. The platform is composed of a microfluidic filtering device and a superhydrophobic surface integrating surface enhanced Raman scattering (SERS) sensors. The microfluidic device was used to filter specific peptides (MW 1553.73 D) derived from the BRCA1 protein, a tumor-suppressor molecule which plays a pivotal role in the development of breast cancers, from albumin (66.5 KD), the most represented protein in human plasma. The filtering process consisted of driving the complex mixture through a porous membrane having a cut-off of 12–14 kD by hydrodynamic flow. The filtered samples coming out of the microfluidic device were subsequently deposited on a superhydrophobic surface formed by micro pillars on top of which nanograins were fabricated. The nanograins coupled to a Raman spectroscopy instrument acted as a SERS sensor and allowed analysis of the filtered sample on top of the surface once it evaporated. By using the presented platform, we demonstrate being able to sort small peptides from bigger proteins and to detect them by using a label-free technique at a resolution down to 0.1 ng μL−1. The combination of microfluidics and nanotechnology to develop the presented microfluidic platform may give rise to a new generation of biosensors capable of detecting low concentration samples from complex mixtures without the need for any sample pretreatment or labelling. The developed devices could have future applications in the field of early diagnosis of severe illnesses, e.g. early cancer detection.
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- 2014
80. pEGFR-Tyr 845 expression as prognostic factors in oral squamous cell carcinoma
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Lorenzo Lo Muzio, Valentina Cozza, Carolina Sbordone, Michele Caraglia, Alfredo De Rosa, Giuseppina Liguori, Giovanni Cuda, Marina Di Domenico, Gianluca Florio, Giuseppe Pannone, Simona Losito, Francesco Longo, Stefania Staibano, Anna Grimaldi, Angela Santoro, Gerardo Botti, Gabriella Aquino, Marilena Mattoni, Pantaleo Bufo, Rosario Serpico, Renato Franco, Aquino, G, Pannone, G, Santoro, A, Liguori, G, Franco, Renato, Serpico, R, Florio, G, DE ROSA, Alfredo, Mattoni, M, Cozza, V, Botti, G, Losito, S, Longo, F, Staibano, S, Cuda, G, Lo Muzio, L, Sbordone, C, Bufo, P, Grimaldi, A, Caraglia, Michele, DI DOMENICO, Marina, Franco, R, De Rosa, A, Staibano, Stefania, Caraglia, M, and Di Domenico, M.
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Adult ,Male ,Cancer Research ,Pathology ,medicine.medical_specialty ,EGFR ,medicine.medical_treatment ,Targeted therapy ,Phosphorylated EGFR ,medicine ,Carcinoma ,Humans ,Epidermal growth factor receptor ,Phosphorylation ,Grading (tumors) ,Aged ,Neoplasm Staging ,Retrospective Studies ,Aged, 80 and over ,Pharmacology ,Mouth neoplasm ,Paraffin Embedding ,Tissue microarray ,biology ,medicine.diagnostic_test ,Middle Aged ,Prognosis ,medicine.disease ,Immunohistochemistry ,ErbB Receptors ,stomatognathic diseases ,Oncology ,Tissue Array Analysis ,Carcinoma, Squamous Cell ,Clinical Study ,Cancer research ,biology.protein ,Tyrosine ,Molecular Medicine ,Female ,Mouth Neoplasms ,OSCC ,Fluorescence in situ hybridization - Abstract
The EGFR (epidermal growth factor receptor) a member of the family of transmembrane protein kinase receptors known as the erbB family shows a significant correlation with the presence of metastases and poorly differentiated oral cancer. Aim of the present work is to define the key-role of EGFR in oral cancer prognosis. We have analyzed the EGFR expression on 149 cases of oral squamous cell cancers (OSCC) and we have found that it was poorly expressed in normal oral epithelium, but its expression was significantly increased in OSCCs. Moreover, we have recorded that both pEGFR-Tyr 845 and pEGFR-Tyr 1068 were mainly distributed in high histological grading and in advanced stages. Western blotting has confirmed the total absence of EGFR phosphorylation in normal oral epithelium and the higher level of protein phosphorylation in representative cases of OSCCs. The EGF-R amplification was found by fluorescence in situ hybridization (FISH) in 14% of OSCC; interestingly, EGF-R amplification was mainly observed in OSCC with higher histological grading (G2 and G3) and advanced stage (pT4) sub-groups. Kaplan-Meyer survival analysis suggested that patients with positive pEGFR-Tyr 845 tumors had a worse prognosis and were bad responders to chemotherapy. These results confirm the central role of EGF-R activation status as a prognostic biomarker in OSCC The EGFR (epidermal growth factor receptor) a member of the family of transmembrane protein kinase receptors known as the erbB family shows a significant correlation with the presence of metastases and poorly differentiated oral cancer. Aim of the present work is to define the key-role of EGFR in oral cancer prognosis. We have analyzed the EGFR expression on 149 cases of oral squamous cell cancers (OSCC) and we have found that it was poorly expressed in normal oral epithelium, but its expression was significantly increased in OSCCs. Moreover, we have recorded that both pEGFR-Tyr 845 and pEGFR-Tyr 1068 were mainly distributed in high histological grading and in advanced stages. Western blotting has confirmed the total absence of EGFR phosphorylation in normal oral epithelium and the higher level of protein phosphorylation in representative cases of OSCCs. The EGF-R amplification was found by fluorescence in situ hybridization (FISH) in 14% of OSCC; interestingly, EGF-R amplification was mainly observed in OSCC with higher histological grading (G2 and G3) and advanced stage (pT4) sub-groups. Kaplan-Meyer survival analysis suggested that patients with positive pEGFR-Tyr 845 tumors had a worse prognosis and were bad responders to chemotherapy. These results confirm the central role of EGF-R activation status as a prognostic biomarker in OSCC. © 2012 Landes Bioscience.
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- 2012
81. Breaking the diffusion limit with super-hydrophobic delivery of molecules to plasmonic nanofocusing SERS structures
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Remo Proietti Zaccaria, Francesco Gentile, Gheorghe Cojoc, Federico Mecarini, Roberto Cingolani, Andrea Toma, Carlo Liberale, E. Di Fabrizio, F. De Angelis, Maria Laura Coluccio, Gerardo Perozziello, Gobind Das, Giovanni Cuda, Manola Moretti, Patrizio Candeloro, Luca Tirinato, Angelo Accardo, De Angelis, F., Gentile, Francesco, Mecarini, F., Das, G., Moretti, M., Candeloro, P., Coluccio, M. L., Cojoc, G., Accardo, A., Liberale, C., Zaccaria, R. P., Perozziello, G., Tirinato, L., Toma, A., Cuda, G., Cingolani, R., and Di Fabrizio, E.
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Materials science ,Electronic, Optical and Magnetic Material ,Evaporation ,Nanotechnology ,02 engineering and technology ,010402 general chemistry ,021001 nanoscience & nanotechnology ,01 natural sciences ,Atomic and Molecular Physics, and Optics ,0104 chemical sciences ,Electronic, Optical and Magnetic Materials ,symbols.namesake ,symbols ,Molecule ,Diffusion limit ,0210 nano-technology ,Spectroscopy ,Raman spectroscopy ,Plasmon - Abstract
The detection of a few molecules in a highly diluted solution is of paramount interest in fields including biomedicine, safety and eco-pollution in relation to rare and dangerous chemicals. Nanosensors based on plasmonics are promising devices in this regard, in that they combine the features of high sensitivity, label-free detection and miniaturization. However, plasmonic-based nanosensors, in common with general sensors with sensitive areas on the scale of nanometres, cannot be used directly to detect molecules dissolved in femto- or attomolar solutions. In other words, they are diffusion-limited and their detection times become impractical at such concentrations. In this Article, we demonstrate, by combining super-hydrophobic artificial surfaces and nanoplasmonic structures, that few molecules can be localized and detected even at attomolar (10−18 mol l−1) concentration. Moreover, the detection can be combined with fluorescence and Raman spectroscopy, such that the chemical signature of the molecules can be clearly determined.
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- 2011
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82. Bilateral cataract in a subject carrying a C to A transition in the L ferritin promoter region
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Antonia Nisticò, Maddalena Di Sanzo, Giovanni Cuda, Annalisa Fregola, Francesco Costanzo, Michela Grosso, Barbara Quaresima, Maria Concetta Faniello, Faniello, M. C., Di Sanzo, M., Quaresima, B., Nisticò, A., Fregola, A., Grosso, Michela, Cuda, G., and Costanzo, F.
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Bilateral ctaract ,Clinical Biochemistry ,medicine.disease_cause ,Polymerase Chain Reaction ,Polymorphism, Single Nucleotide ,Cataract ,Pathogenesis ,Transcriptional regulation ,medicine ,Humans ,Promoter Regions, Genetic ,Gene ,Ferritin ,Mutation ,biology ,Transition (genetics) ,Promoter ,General Medicine ,Molecular biology ,Real-time polymerase chain reaction ,Italy ,Gene promoter ,Ferritins ,biology.protein ,HeLa Cells - Abstract
Objectives The aim of this study is to evaluate the potential impact of mutations in the promoter region of the L ferritin gene on its transcriptional activity. Design and methods To search for the presence of mutations in the promoter of the L gene, we amplified by PCR a DNA region of about 385 n.t. in 100 healthy subjects from Southern Italy. Results A subject carrying a C to A transition in position − 216 was identified. This transition causes an increased transcriptional activity in vitro. This finding was substantiated by Real Time Quantitative PCR, which showed increased levels of L ferritin mRNA. Conclusions A previously unidentified mutation in the promoter region of the L ferritin gene was detected in an individual. Interestingly, this subject is affected by bilateral cataract, a disease that has been correlated, in a subset of patients, with high levels of circulating ferritin. We hypothesize that changes in the expression of the L ferritin might be linked, at least to a certain extent, to the pathogenesis of this rare eye disease.
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- 2009
83. Mechanical Stress Downregulates MHC Class I Expression on Human Cancer Cell Membrane
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Rosanna, La Rocca, Rossana, Tallerico, Almosawy, Talib Hassan, Gobind, Das, Tadepally, Lakshmikanth, Lakshmikanth, Tadepally, Marco, Matteucci, Carlo, Liberale, Maria, Mesuraca, Domenica, Scumaci, Francesco, Gentile, Gheorghe, Cojoc, Gerardo, Perozziello, Antonio, Ammendolia, Adriana, Gallo, Klas, Kärre, Giovanni, Cuda, Patrizio, Candeloro, Enzo, Di Fabrizio, Ennio, Carbone, La Rocca, Rosanna, Tallerico, Rossana, Hassan, Almosawy Talib, Das, Gobind, Tadepally, Lakshmikanth, Matteucci, Marco, Liberale, Carlo, Mesuraca, Maria, Scumaci, Domenica, Gentile, Francesco, Cojoc, Gheorghe, Perozziello, Gerardo, Ammendolia, Antonio, Gallo, Adriana, Kärre, Kla, Cuda, Giovanni, Candeloro, Patrizio, Di Fabrizio, Enzo, Carbone, Ennio, La Rocca, R, Tallerico, R, Talib Hassan, A, Das, G, Tadepally, L, Matteucci, M, Liberale, C, Mesuraca, M, Scumaci, D, Gentile, F, Cojoc, G, Perozziello, G, Ammendolia, A, Gallo, A, Kärre, K, Cuda, G, Di Fabrizio, E, Candeloro, P, and Carbone, E
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Lymphocyte ,Spectrum Analysis, Raman ,Cell-Mediated Immunity ,Spectrum Analysis Techniques ,HEK293 Cell ,Neoplasms ,Medicine and Health Sciences ,Cytotoxic T cell ,Cells, Cultured ,Multidisciplinary ,biology ,Medicine (all) ,Physics ,Classical Mechanics ,Cell biology ,Killer Cells, Natural ,medicine.anatomical_structure ,Phenotype ,Physical Sciences ,Medicine ,Mechanical Stress ,Human ,Research Article ,Tumor Immunology ,Raman Spectroscopy ,Science ,Immunology ,Down-Regulation ,Major histocompatibility complex ,Research and Analysis Methods ,SDG 3 - Good Health and Well-being ,MHC class I ,medicine ,Humans ,Antigen-presenting cell ,Biochemistry, Genetics and Molecular Biology (all) ,Cell growth ,MHC Class I Gene ,Cell Membrane ,Histocompatibility Antigens Class I ,Immunity ,Correction ,Biology and Life Sciences ,HEK293 Cells ,Agricultural and Biological Sciences (all) ,Cancer cell ,biology.protein ,Neoplasm ,Tumor Escape ,Clinical Immunology ,Stress, Mechanical - Abstract
In our body, cells are continuously exposed to physical forces that can regulate different cell functions such as cell proliferation, differentiation and death. In this work, we employed two different strategies to mechanically stress cancer cells. The cancer and healthy cell populations were treated either with mechanical stress delivered by a micropump (fabricated by deep X-ray nanolithography) or by ultrasound wave stimuli. A specific down-regulation of Major Histocompatibility Complex (MHC) class I molecules expression on cancer cell membrane compared to different kinds of healthy cells (fibroblasts, macrophages, dendritic and lymphocyte cells) was observed, stimulating the cells with forces in the range of nano-newton, and pressures between 1 and 10 bar (1 bar5100.000 Pascal), depending on the devices used. Moreover, Raman spectroscopy analysis, after mechanical treatment, in the range between 700–1800 cm-1, indicated a relative concentration variation of MHC class I. PCA analysis was also performed to distinguish control and stressed cells within different cell lines. These mechanical induced phenotypic changes increase the tumor immunogenicity, as revealed by the related increased susceptibility to Natural Killer (NK) cells cytotoxic recognition.
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- 2014
84. An alternative model of H ferritin promoter transactivation by c-Jun
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Faniello, Maria C, Chirico, Giuseppa, Quaresima, Barbara, Cuda, Giovanni, Allevato, Giovanna, Bevilacqua, Maria A, Baudi, Francesco, Colantuoni, Vittorio, Cimino, Filiberto, Venuta, Salvatore, Avvedimento, Vittorio E, Costanzo, Francesco, Faniello, Mc, Chirico, G, Quaresima, B, Cuda, G, Allevato, G, Bevilacqua, MARIA ASSUNTA, Baudi, F, Colantuoni, V, Cimino, F, Venuta, S, Avvedimento, Ve, and Costanzo, F.
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Chloramphenicol O-Acetyltransferase ,Transcriptional Activation ,Binding Sites ,Transcription, Genetic ,Proto-Oncogene Proteins c-jun ,Recombinant Fusion Proteins ,Blotting, Western ,Nuclear Proteins ,Cell Biology ,Transfection ,Precipitin Tests ,Biochemistry ,Recombinant Proteins ,Cell Line ,Protein Structure, Tertiary ,Ferritins ,Cyclic AMP ,Trans-Activators ,Humans ,Promoter Regions, Genetic ,Molecular Biology ,Research Article ,HeLa Cells ,Protein Binding - Abstract
c-Jun is a member of the activator protein 1 family, and its interaction with different nuclear factors generates a wide spectrum of complexes that regulate transcription of different promoters. H ferritin promoter transcription is tightly dependent on nuclear factor Y (NFY). Ferritin transcription is activated by c-Jun, although the promoter does not contain a canonical binding site. NFY, on the other hand, does not bind c-Jun in vitro, whereas in vivo c-Jun is found in the complex containing NFY. Moreover, a c-Jun—GCN4 chimaeric construct containing only the transactivation domain of Jun and the basic-region leucine-zipper domain of GCN4 stimulates the H ferritin promoter. A synthetic GAL4 promoter and the cognate activator, the fusion protein NFY—GAL4, are potently activated by c-Jun. Titration of p300 by co-expressing E1A abolishes the stimulatory effect. Moreover, another p300-dependent promoter, the cAMP-response element, can be superactivated by c-Jun using the same mechanism. These data indicate that c-Jun, when activated or overexpressed, is recruited to the H ferritin promoter by p300, which links NFY, bound to DNA, to the complex. These results add a new level of complexity to transcriptional regulation by c-Jun, which can activate p300-dependent promoters without binding directly to the target DNA.
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- 2002
85. Detection of microsatellite instability and loss of heterozygosity in serum DNA of small and non-small cell lung cancer patients: a tool for early diagnosis?
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Piero S Tagliaferri, A. Gallelli, Vito Barbieri, Carmelindo M.E Tranfa, Pierfrancesco Tassone, Giovanni Cuda, Antonia Nisticò, Salvatore Venuta, Francesco Costanzo, Cuda, G, Gallelli, A, Nistico, A, Tassone, P, Barbieri, V, Tagliaferri, P, Costanzo, F, Tranfa, Carmelindo Mario Enrico, and Venuta, S.
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Pulmonary and Respiratory Medicine ,Cancer Research ,Pathology ,medicine.medical_specialty ,business.industry ,Respiratory disease ,Microsatellite instability ,medicine.disease ,Loss of heterozygosity ,Oncology ,Lung disease ,medicine ,Cancer research ,Microsatellite ,Non small cell ,Lung cancer ,business ,Serum dna - Published
- 2000
86. Identification of H ferritin-dependent and independent genes in K562 differentiating cells by targeted gene silencing and expression profiling
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Maddalena Di Sanzo, Heather M. Bond, Giuseppe Viglietto, Giorgio Giurato, Carlo Cosentino, Domenica Scumaci, Giovanni Cuda, Claudia Stellato, Giovanni Morrone, Francesco Romeo, Alessandro Weisz, R. Misaggi, Francesco Costanzo, Maria Concetta Faniello, Tullio Barni, Barbara Quaresima, Francesco Amato, Misaggi, R., Di Sanzo, M., Cosentino, C., Bond, H. M., Scumaci, D., Romeo, F., Stellato, C., Giurato, G., Weisz, A., Quaresima, B., Barni, T., Amato, F., Viglietto, G., Morrone, G., Cuda, G., Faniello, M. C., and Costanzo, F.
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Cellular differentiation ,Microarray ,Biology ,Ferritin Light Chain ,Small hairpin RNA ,shRNA ,RNA interference ,quantitative Real Time Polymerase Chain Reaction ,Gene expression ,K562 Cell ,Genetics ,Cluster Analysis ,Humans ,short hairpin RNA ,Gene silencing ,Gene Regulatory Networks ,Neoplastic transformation ,Gene Silencing ,cardiovascular diseases ,Melanoma cells ,Protein Subunit ,Ferritin ,Cluster Analysi ,Gene Regulatory Network ,Gene Expression Regulation, Leukemic ,Gene Expression Profiling ,Computational Biology ,Cell Differentiation ,General Medicine ,Molecular biology ,FLC ,Gene expression profiling ,Ferritin light chain ,Protein Subunits ,qPCR ,Differentiation ,Ferritins ,Hemin ,RNA Interference ,K562 Cells ,FHC ,Ferritin Heavy Chain ,Human ,Signal Transduction - Abstract
Ferritin is best known as the key molecule in intracellular iron storage, and is involved in several metabolic processes such as cell proliferation, differentiation and neoplastic transformation. We have recently demonstrated that the shRNA silencing of the ferritin heavy subunit (FHC) in a melanoma cell line is accompanied by a consistent modification of gene expression pattern leading to a reduced potential in terms of proliferation, invasiveness, and adhesion ability of the silenced cells. In this study we sought to define the repertoire of genes whose expression might be affected by FHC during the hemin-induced differentiation of the erythromyeloid cell line K562. To this aim, gene expression profiling was performed in four different sets of cells: i) wild type K562; ii) sh-RNA FHC-silenced K562; iii) hemin-treated wild-type K562; and iv) hemin-treated FHC-silenced K562. Statistical analysis of the gene expression data, performed by two-factor ANOVA, identified three distinct classes of transcripts: a) Class 1, including 657 mRNAs whose expression is modified exclusively during hemin-induced differentiation of K562 cells, independently from the FHC relative amounts; b) Class 2, containing a set of 70 mRNAs which are consistently modified by hemin and FHC-silencing; and c) Class 3, including 128 transcripts modified by FHC-silencing but not by hemin. Our data indicate that FHC may function as a modulator of gene expression during erythroid differentiation and add new findings to the knowledge of the complex gene network modulated during erythroid differentiation.
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- 2013
87. Biomarker discovery by plasma proteomics in familial Brugada Syndrome
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M. Di Domenico, Vincenzo Lorenzo Pascali, C Romano Carratelli, S Grasso, P. Ricci, Domenica Scumaci, Annamaria Chiara Santini, Giovanni Cuda, D Conti, Tomei, Carmela Ricciardi, C Di Nunzio, Francesco Ausania, R La Montagna, Ciro Indolfi, FA Rizzo, Francesco Costanzo, Marco Gaspari, Malafoglia, Monica Lamberti, Antonio Oliva, Antonio Curcio, DI DOMENICO, Marina, Scumaci, D, Grasso, S, Gaspari, M, Curcio, A, Oliva, Adriana, Ausania, Af, Di Nunzio, C, Ricciardi, C, Santini, Mario, Rizzo, Antonietta, Romano Carratelli, C, Lamberti, Monica, Conti, D, La Montagna, R, Tomei, V, Malafoglia, V, Pascali, Vl, Ricci, P, Indolfi, C, Costanzo, F, and Cuda, G.
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Male ,Apolipoprotein E ,Proteomics ,medicine.medical_specialty ,Proteome ,Antithrombin III ,Gene mutation ,Fibrinogen ,Sudden death ,NAV1.5 Voltage-Gated Sodium Channel ,Electrocardiography ,Apolipoproteins E ,Tandem Mass Spectrometry ,Internal medicine ,Mass spectrometry Send ,Medicine ,Humans ,Electrophoresis, Gel, Two-Dimensional ,Brugada syndrome ,Biomarker discovery ,Clusterin ,biology ,Mass spectrometry ,business.industry ,Serum proteomics ,Biomarker ,Settore MED/43 - MEDICINA LEGALE ,Early diagnosi ,medicine.disease ,Early diagnosis ,Pedigree ,Surgery ,Endocrinology ,alpha 1-Antitrypsin ,biology.protein ,Female ,Prothrombin ,business ,Protein Processing, Post-Translational ,SUDDEN CARDIAC DEATH ,Biomarkers ,medicine.drug - Abstract
Brugada Syndrome (BS) is a polygenic inherited cardiac disease characterized by life-threatening arrhythmias and high incidence of sudden death. In this study, two-dimensional gel electrophoresis (2D-PAGE) coupled to mass spectrometry (LC-MS/MS) was used to investigate specific changes in the plasma proteome of BS patients and family members sharing the same gene mutation (SCN5AQ1118X), with the aim to identify novel disease biomarkers. Our data demonstrate that the levels of several proteins were significantly altered in BS patients compared with controls. In particular, apolipoprotein E, prothrombin, vitronectin, complement-factor H, vitamin-D-binding protein, voltage-dependent anion-selective channel protein 3 and clusterin were considerably increased in plasma sample of BS patients, whereas alpha-1-antitrypsin, fibrinogen and angiotensinogen were considerably decreased; moreover, post-translational modifications of antithrombin-III were detected in all affected individuals. On the light of these results, we hypothesize that these proteins might be considered as potential markers for the identification of disease status in BS.
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- 2013
88. Characterization of breast cancer interstitial fluids by TmT labeling, LTQ-Orbitrap Velos mass spectrometry, and pathway analysis
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Nuria Vadalà, Cinzia Raso, Xuemei Han, John R. Yates, Giovanni Cuda, Sung Kyu Park, M. Renne, Ubaldo Prati, Natalia Malara, Vincenzo Mollace, Carlo Cosentino, Marco Gaspari, Daniel B. McClatchy, Francesco Amato, Raso, C., Cosentino, C., Gaspari, M., Malara, N., Han, X., Mcclatchy, D., Park, S. K., Renne, M., Vadala, N., Prati, U., Cuda, G., Mollace, V., Amato, F., and Yates, J. R.
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Stromal cell ,Proteome ,Breast Neoplasms ,Computational biology ,Biology ,Tandem mass tag ,Bioinformatics ,Orbitrap ,Tandem mass spectrometry ,Biochemistry ,Article ,law.invention ,03 medical and health sciences ,0302 clinical medicine ,Breast cancer ,law ,Phyllodes Tumor ,Tandem Mass Spectrometry ,Protein Interaction Mapping ,medicine ,Humans ,Protein Interaction Maps ,030304 developmental biology ,Epigenomics ,0303 health sciences ,Staining and Labeling ,Carcinoma, Ductal, Breast ,Cancer ,Extracellular Fluid ,General Chemistry ,medicine.disease ,3. Good health ,Molecular Weight ,030220 oncology & carcinogenesis ,Female ,Protein Interaction Map ,Breast Neoplasm ,Human - Abstract
Cancer is currently considered as the end point of numerous genomic and epigenomic mutations and as the result of the interaction of transformed cells within the stromal microenvironment. The present work focuses on breast cancer, one of the most common malignancies affecting the female population in industrialized countries. In this study, we perform a proteomic analysis of bioptic samples from human breast cancer, namely, interstitial fluids and primary cells, normal vs disease tissues, using tandem mass tags (TmT) quantitative mass spectrometry combined with the MudPIT technique. To the best of our knowledge, this work, with over 1700 proteins identified, represents the most comprehensive characterization of the breast cancer interstitial fluid proteome to date. Network analysis was used to identify functionally active networks in the breast cancer associated samples. From the list of differentially expressed genes, we have retrieved the associated functional interaction networks. Many different signaling pathways were found activated, strongly linked to invasion, metastasis development, proliferation, and with a significant cross-talking rate. This pilot study presents evidence that the proposed quantitative proteomic approach can be applied to discriminate between normal and tumoral samples and for the discovery of yet unknown carcinogenesis mechanisms and therapeutic strategies.
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- 2012
89. Proteomics in Ménière disease
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Milena Saccomanno, Ettore Cassandro, Giuseppe Chiarella, Claudia Cassandro, Giovanni Cuda, Barbara Quaresima, Claudio Petrolo, Carmela Ricciardi, Marina Di Domenico, Marco Gaspari, Maria Concetta Faniello, Francesco Costanzo, Domenica Scumaci, Chiarella, G, Saccomanno, M, Scumaci, D, Gaspari, M, Faniello, Mc, Quaresima, B, DI DOMENICO, Marina, Riccardi, C, Petrolo, C, Cassandro, C, Costanzo, F, Cuda, G, and Cassandro, E.
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Electrophoresis ,Male ,Proteomics ,Physiology ,Vitamin D-binding protein ,Clinical Biochemistry ,Fibrinogen ,Mass Spectrometry ,medicine ,Humans ,Electrophoresis, Gel, Two-Dimensional ,blood/genetics/physiopathology ,Endolymphatic hydrops ,Meniere Disease ,Whole blood ,chemistry.chemical_classification ,Chromatography ,Liquid ,Gel ,business.industry ,Albumin ,Cell Biology ,analysis/blood ,Middle Aged ,medicine.disease ,Blood proteins ,chemistry ,Transferrin ,Factor H ,Immunology ,Two-Dimensional ,Biological Markers ,Female ,business ,analysis/blood, Chromatography ,Liquid, Electrophoresis ,Two-Dimensional, Female, Humans, Male, Mass Spectrometry, Meniere Disease ,blood/genetics/physiopathology, Middle Aged, Proteomics ,Biomarkers ,medicine.drug ,Chromatography, Liquid - Abstract
Meniere's disease (MD) is a disorder of the inner ear characterized by an insidious onset and aspecific symptoms, such as dizziness, vertigo, tinnitus, and hearing loss, that may become very debilitating. The presence of endolymphatic hydrops is a common feature in MD patients, but the pathophysiology is still largely unknown. In this study, we have used a proteomics-driven approach to identify potential biomarkers of MD. To this end, plasma was obtained from whole blood of 16 individuals previously diagnosed as suffering from MD and compared to plasma from healthy donors. A depletion of the highly abundant proteins (i.e., albumin, IgG, transferrin, etc.) was performed in order to enhance the chance of detection of the less represented ones, therefore reducing the noise-background. Two-dimensional gel electrophoresis, followed by in-gel tryptic digestion of the selected spots and LC-MS/MS analysis, allowed us to identify a set of proteins whose expression appears to be differentially modulated in patients versus controls. In particular: complement factor H and B, fibrinogen alpha and gamma chains, beta-actin and pigment epithelium derived factor are over expressed; on the other hand, the levels of beta-2 glycoprotein-1, vitamin D binding protein and apolipoprotein-1 are significantly decreased in the plasma of MD-affected individuals. Even though preliminary and not necessarily linked directly to the molecular pathogenesis of the disease, our original findings suggest that a molecular signature, represented by the plasma protein profile previously described, might represent a potentially powerful, innovative and not invasive tool for early diagnosis and clinical management of MD patients. J. Cell. Physiol. 227: 308-312, 2012. © 2011 Wiley Periodicals, Inc.
- Published
- 2011
90. Calpain3 is expressed in a proteolitically active form in papillomavirus-associated urothelial tumors of the urinary bladder in cattle
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Franco Roperto, Gianni Cuda, Monica Averna, Marco Gaspari, Roberta De Tullio, Valeria Russo, Sante Roperto, Cinzia Raso, Giuseppe Borzacchiello, Roberto Stifanese, Orlando Paciello, Roperto, Sante, De Tullio, R, Raso, C, Stifanese, R, Russo, Valeria, Gaspari, M, Borzacchiello, Giuseppe, Averna, M, Paciello, Orlando, Cuda, G, and Roperto, FRANCO PEPPINO
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Urinary system ,Science ,Biology ,medicine.disease_cause ,Virology/Emerging Viral Diseases ,Infectious Diseases/Viral Infections ,medicine ,Animals ,Neoplasms, Glandular and Epithelial ,Urothelium ,Bovine papillomavirus 1 ,Bovine papillomavirus ,Multidisciplinary ,Bladder cancer ,Urinary bladder ,Calpain ,Reverse Transcriptase Polymerase Chain Reaction ,Papillomavirus Infections ,medicine.disease ,biology.organism_classification ,Molecular biology ,Pathology/Molecular Pathology ,Gene Expression Regulation, Neoplastic ,Reverse transcription polymerase chain reaction ,medicine.anatomical_structure ,Urinary Bladder Neoplasms ,E2F3 Transcription Factor ,Virology/Animal Models of Infection ,Immunohistochemistry ,Medicine ,Calcium ,Cattle ,Carcinogenesis ,Research Article ,Virology/Viruses and Cancer - Abstract
BackgroundCalpain 3 (Capn3), also named p94, is a skeletal muscle tissue-specific protein known to be responsible for limb-girdle muscular dystrophy type 2A (LGMD2A). Recent experimental studies have hypothesized a pro-apoptotic role of Capn3 in some melanoma cell lines. So far the link between calpain3 and tumors comes from in vitro studies. The objective of this study was to describe Capn3 activation in naturally occurring urothelial tumors of the urinary bladder in cattle.Methods and findingsHere we describe, for the first time in veterinary and comparative oncology, the activation of Capn3 in twelve urothelial tumor cells of the urinary bladder of cattle. Capn3 protein was initially identified with nanoscale liquid chromatography coupled with tandem mass spectrometry (nano LC-MS/MS) in a co-immunoprecipitation experiment on E2F3, known to be a transcription factor playing a crucial role in bladder carcinogenesis in humans. Capn3 expression was then confirmed by reverse transcription polymerase chain reaction (RT-PCR). Finally, the Ca(2+)-dependent proteolytic activity of Capn3 was assayed following ion exchange chromatography. Morphologically, Capn3 expression was documented by immunohistochemical methods. In fact numerous tumor cells showed an intracytoplasmic immunoreactivity, which was more rarely evident also at nuclear level. In urothelial tumors, bovine papillomavirus type 2 (BPV-2) DNA was amplified by PCR and the expression of E5 protein, the major oncogenic protein of BVP-2, was detected by western blotting, immunohistochemistry, and immunofluorescence. E2F3 overexpression and pRb protein downregulation were shown by western blotting.ConclusionThe role of capn3 protein in urothelial cancer of the urinary bladder remains to be elucidated: further studies would be required to determine the precise function of this protease in tumor development and progression. However, we suggest that activated Capn3 may be involved in molecular pathways leading to the overexpression of E2F3, which in turn could be responsible for urothelial tumor cell proliferation also in cattle, though other mechanisms are likely to exist. If further studies corroborate the important role of Capn3 in urothelial tumors of the urinary bladder, cattle with urinary tumors may prove useful as animal model for bladder carcinogenesis.
- Published
- 2010
91. p53-Mediated downregulation of H ferritin promoter transcriptional efficiency via NF-Y
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Francesco Costanzo, Barbara Quaresima, Giovanni Spinelli, Francesco Baudi, Salvatore Venuta, Giovanni Morrone, Maddalena Di Sanzo, Maria Concetta Faniello, Giannino Del Sal, Giovanni Cuda, Valentina Di Caro, Faniello, Mc, DI SANZO, M, Quaresima, B, Baudi, F, DI CARO, V, Cuda, G, Morrone, G, DEL SAL, Giannino, Spinelli, G, Venuta, S, Costanzo, F., Faniello, C, Di Sanzo, M, Di Caro, V, Del Sal, G, and Costanzo, F
- Subjects
Chromatin Immunoprecipitation ,Multiprotein complex ,Transcription, Genetic ,Down-Regulation ,Biology ,Biochemistry ,Transcriptional regulation ,Downregulation and upregulation ,Transcription (biology) ,Ferritin gene ,Humans ,Electrophoretic mobility shift assay ,p300-CBP Transcription Factors ,Promoter Regions, Genetic ,Transcription factor ,Gene ,Transcriptional factor ,Cell Biology ,HCT116 Cells ,Molecular biology ,Gene Expression Regulation, Neoplastic ,CCAAT-Binding Factor ,Doxorubicin ,Apoferritins ,Tumor Suppressor Protein p53 ,Chromatin immunoprecipitation ,HeLa Cells ,Protein Binding - Abstract
The tumor suppressor protein p53 triggers many of the cellular responses to DNA damage by regulating the transcription of a series of downstream target genes. p53 acts on the promoter of the target genes by interacting with the trimeric transcription factor NF-Y. H ferritin promoter activity is tightly dependent on a multiprotein complex called Bbf; on this complex NF-Y plays a major role. The aim of this work was to study the modulation of H ferritin expression levels by p53. CAT reporter assays indicate that: (i) p53 overexpression strongly downregulates the transcriptional efficiency driven by an H ferritin promoter construct containing only the NF-Y recognition sequence and that the phenomenon is reverted by p53 siRNA; (ii) the p53 C-terminal region is sufficient to elicitate this regulation and that a correct C-terminal acetylation is also required. The H ferritin promoter displays no p53-binding sites; chromatin immunoprecipitation assays indicate that p53 is recruited on this promoter by NF-Y. The p53–NF-Y interaction does not alter the NF-Y DNA-binding ability as indicated by electrophoretic mobility shift assay (EMSA) analysis. These results demonstrate that the gene coding for the H ferritin protein belongs to the family of p53-regulated genes, therefore adding a new level of complexity to the regulation of the H ferritin transcription and delineate a role for this protein in a series of cellular events triggered by p53 activation.
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- 2008
92. Endothelin-1 induces proliferation of human lung fibroblasts and IL-11 secretion through an ET(A) receptor-dependent activation of MAP kinases
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Luca Gallelli, Marilisa De Nardo, Carlo Vancheri, Francesco Costanzo, Mario Caputi, Mauro Maniscalco, Giovanni Cuda, Bruno D'Agostino, V. Gioffrè, C. Mastruzzo, Serafino A. Marsico, Matteo Sofia, Francesco Rossi, Girolamo Pelaia, Alessandro Vatrella, Umberto Galderisi, Elisa Trovato Salinaro, Nunzio Crimi, Rosario Maselli, D. Fratto, Gallelli, Luca, Pelaia, Girolamo, D'Agostino, Bruno, Cuda, Giovanni, Vatrella, Alessandro, Fratto, Donatella, Gioffrè, Vincenza, Galderisi, Umberto, De Nardo, Marilisa, Mastruzzo, Claudio, Salinaro, Elisa Trovato, Maniscalco, Mauro, Sofia, Matteo, Crimi, Nunzio, Rossi, Francesco, Caputi, Mario, Costanzo, Francesco S, Maselli, Rosario, Marsico, Serafino A, Vancheri, Carlo, Gallelli, L, Pelaia, G, Cuda, G, Vatrella, A, Fratto, D, Gioffre, V, DE NARDO, M, Mastruzzo, C, Salinaro, Et, Maniscalco, M, Sofia, M, Crimi, N, Rossi, F, Caputi, M, Costanzo, F, Maselli, R, Marsico, Sa, and Vancheri, C.
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MAPK/ERK pathway ,Time Factors ,Cell Survival ,Endothelin A Receptor Antagonists ,Biology ,Mitogen-Activated Protein Kinase ,Biochemistry ,Endothelin B Receptor Antagonist ,Humans ,Secretion ,RNA, Messenger ,Phosphorylation ,Receptor ,Molecular Biology ,Lung ,Cell Proliferation ,IL-6 ,Endothelin-1 ,Kinase ,Cell growth ,Interleukin-6 ,lung fibroblasts ,Cell Biology ,ET-1 receptors ,Fibroblasts ,Interleukin-11 ,Receptor, Endothelin A ,Endothelin 1 ,MAPK ,Receptor, Endothelin B ,ET-1 ,Cell biology ,Endothelin B Receptor Antagonists ,Intracellular signal transduction ,IL-11 ,Cell culture ,Fibroblast ,Mitogen-Activated Protein Kinases ,Endothelin A Receptor Antagonist ,Human - Abstract
Endothelin-1 (ET-1) is implicated in the fibrotic responses characterizing interstitial lung diseases, as well as in the airway remodeling process occurring in asthma. Within such a context, the aim of our study was to investigate, in primary cultures of normal human lung fibroblasts (NHLFs), the ET-1 receptor subtypes, and the intracellular signal transduction pathways involved in the proliferative effects of this peptide. Therefore, cells were exposed to ET-1 in the presence or absence of an overnight pre-treatment with either ETA or ETB selective receptor antagonists. After cell lysis, immunoblotting was performed using monoclonal antibodies against the phosphorylated, active forms of mitogen-activated protein kinases (MAPK). ET-1 induced a significant increase in MAPK phosphorylation pattern, and also stimulated fibroblast proliferation and IL-6/IL-11 release into cell culture supernatants. All these effects were inhibited by the selective ETA antagonist BQ-123, but not by the specific ETB antagonist BQ-788. The stimulatory influence of ET-1 on IL-11, but not on IL-6 secretion, was prevented by MAPK inhibitors. Therefore, such results suggest that in human lung fibroblasts ET-1 exerts a profibrogenic action via an ETA receptor-dependent, MAPK-mediated induction of IL-11 release and cell proliferation. © 2005 Wiley-Liss, Inc.
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- 2005
93. Mitogen-activated protein kinases and asthma
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Alberto Abbruzzese, Francesco Costanzo, Michele Caraglia, Girolamo Pelaia, Luca Gallelli, Mario Caputi, Rosario Maselli, M. Marra, Alessandro Vatrella, Giovanni Cuda, Serafino A. Marsico, Pelaia, G., Cuda, G., Vatrella, S., Gallelli, L., Caraglia, Michele, Marra, M., ABBRUZZESE SACCARDI, A., Caputi, M., Maselli, R., Costanzo, F., and Marsico, S. A.
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MAPK/ERK pathway ,MAP Kinase Signaling System ,Physiology ,T cell ,Clinical Biochemistry ,MAPK ,asthma ,Bronchi ,Biology ,Anti-asthmatic Agent ,Adrenal Cortex Hormones ,medicine ,Extracellular ,Humans ,Anti-Asthmatic Agents ,Enzyme Inhibitors ,Map KINASE ,Molecular Structure ,Kinase ,Cell Biology ,Eosinophil ,medicine.disease ,respiratory tract diseases ,Cell biology ,Enzyme Activation ,medicine.anatomical_structure ,Bronchial hyperresponsiveness ,Immune System ,Immunology ,Mitogen-Activated Protein Kinases ,Signal transduction - Abstract
Mitogen-activated protein kinases (MAPKs) are evolutionary conserved enzymes which play a key role in signal transduction mediated by cytokines, growth factors, neurotransmitters and various types of environmental stresses. In the airways, these extracellular stimuli elicit complex inflammatory and structural changes leading to the typical features of asthma including T cell activation, eosinophil and mast cell infiltration, as well as bronchial hyperresponsiveness and airway remodelling. Because MAPKs represent an important point of convergence for several different signalling pathways, they affect multiple aspects of normal airway function and also significantly contribute to asthma pathophysiology. Therefore, this review focuses on the crucial involvement of MAPKs in asthma pathogenesis, thus also discussing their emerging role as molecular targets for anti-asthma drugs.
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- 2005
94. Mitogen-activated protein kinases: new molecular targets for pharmacological treatment of inflammatory lung diseases
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Girolamo Pelaia, Sa Marsico, Alessandro Vatrella, D. Fratto, Rosario Maselli, Giovanni Cuda, Pierosandro Tagliaferri, Francesco Costanzo, Pelaia, G., Cuda, G., Vatrella, Alessandro, Fratto, D., Tagliaferri, P., Maselli, R., Costanzo, F. S., and Marsico, S. A.
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Pharmacology ,Lung ,Kinase ,business.industry ,Extracellular signal-regulated kinases ,Mitogen-activated protein ,Immunology ,Pharmacological treatment ,medicine.anatomical_structure ,Molecular targets ,medicine ,Cancer research ,Immunology and Allergy ,ASK1 ,business - Published
- 2003
95. Effects of transfoming growth factor-beta and budesonide on mitogen-activated protein kinase activation and apoptosis in airway epithelial cells
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D. Fratto, Girolamo Pelaia, Rosa Daniela Grembiale, Francesco Costanzo, Rosario Maselli, Giovanni Cuda, Pierosandro Tagliaferri, Alessandro Vatrella, Serafino A. Marsico, Pelaia, G., Cuda, G., Vatrella, Alessandro, Fratto, D., Grembiale, R. D., Tagliaferri, P., Maselli, R., Costanzo, F. S., and Marsico, S. A.
- Subjects
Pulmonary and Respiratory Medicine ,MAPK/ERK pathway ,Programmed cell death ,budesonide ,Pyridines ,p38 mitogen-activated protein kinases ,Administration, Topical ,Clinical Biochemistry ,Anti-Inflammatory Agents ,Bronchi ,Apoptosis ,airway epithelial cell ,chemistry.chemical_compound ,Transforming Growth Factor beta ,Humans ,Propidium iodide ,Enzyme Inhibitors ,Phosphorylation ,tgf-beta ,MAPK ,Molecular Biology ,Glucocorticoids ,Cells, Cultured ,Anthracenes ,Flavonoids ,biology ,Kinase ,Caspase 3 ,Imidazoles ,Epithelial Cells ,Cell Biology ,Transforming growth factor beta ,Cell biology ,Enzyme Activation ,chemistry ,Mitogen-activated protein kinase ,Caspases ,biology.protein ,Signal transduction ,Mitogen-Activated Protein Kinases ,Signal Transduction - Abstract
Airway epithelial cells play a central role in the inflammatory, apoptotic, and remodeling processes associated with asthma. Within this context, a key function is exerted by transforming growth factor-beta (TGF-beta), whose biological effects are mediated at least in part by mitogen-activated protein kinases (MAPKs). The aim of our study was to investigate, in primary cultures of human bronchial epithelial cells (HBEC), the effects of TGF-beta (10 ng/ml) on both MAPK activation and apoptosis, in the presence or absence of a pretreatment with budesonide (10-8 M). MAPK activation was detected by Western blotting, using anti-phospho-MAPK monoclonal antibodies, which specifically recognize the phosphorylated, active forms of these enzymes. Apoptosis was assayed by caspase-3 activation and fluorescence microscopy, using annexin-V (An-V) and propidium iodide (PI) as markers of cell death. Our results show that TGF-beta induced a marked ( reverse similar 9-fold) increase in p38 MAPK phosphorylation, and also dramatically enhanced cell death, which was completely prevented by specific MAPK inhibitors. Both MAPK activation and apoptosis were effectively inhibited by budesonide (BUD), thereby suggesting that the powerful antiapoptotic action of inhaled glucocorticoids may be very important for their protective role against epithelial injury, which represents a key pathogenic event in asthma.
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- 2003
96. New possible role of statins in age-related diseases
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Mariarosaria Santillo, Giovanni Cuda, Antonio Ruocco, Alfredo Postiglione, Enrico V. Avvedimento, Rosalba Serù, Roberto Paternò, Ruocco, A, Postiglione, A, Santillo, Mariarosaria, Seru', R, Avvedimento, Ev, Cuda, G, and Paterno', Roberto
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Gerontology ,Apoptosis ,Coronary Disease ,Pharmacology ,Coronary disease ,In Vitro Techniques ,Rats sprague dawley ,Rats, Sprague-Dawley ,Age related ,medicine ,Dementia ,Animals ,Humans ,Stroke ,Serpins ,business.industry ,Blood Proteins ,medicine.disease ,Blood proteins ,Rats ,Hydroxymethylglutaryl-CoA Reductase Inhibitors ,Geriatrics and Gerontology ,business - Published
- 2002
97. Protection of human endothelial cells from oxidative stress: role of Ras-ERK1/2 signaling
- Author
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Giovanni Cuda, Roberto Ceravolo, Maurizio Bifulco, Filippo Schepis, Enrico V. Avvedimento, Mariarosaria Santillo, M. Candigliota, Roberto Paternò, Evelina Mele, Antonio Ruocco, Nicola Perrotti, Susanna Cassano, Francesco Perticone, Maria Concetta Faniello, Cuda, G, Paterno', Roberto, Ceravolo, R, Candigliota, M, Perrotti, N, Perticone, F, Faniello, Mc, Schepis, F, Ruocco, A, Mele, E, Cassano, S, Bifulco, M, Santillo, Mariarosaria, and Avvedimento, VITTORIO ENRICO
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MAPK/ERK pathway ,Endothelium ,Mitogen-Activated Protein Kinase 3 ,Protein Prenylation ,Apoptosis ,Oncogene Protein p21(ras) ,Biology ,Transfection ,medicine.disease_cause ,Structure-Activity Relationship ,Methionine ,Physiology (medical) ,Anti-apoptotic Ras signalling cascade ,medicine ,Humans ,Protein Isoforms ,Small GTPase ,Enzyme Inhibitors ,human endothelial cells ,oxidative stress ,Ras-ERK1/2 signaling ,Cells, Cultured ,Genes, Dominant ,Mitogen-Activated Protein Kinase 1 ,chemistry.chemical_classification ,Reactive oxygen species ,Hydrogen Peroxide ,Cell biology ,Endothelial stem cell ,Oxidative Stress ,Genes, ras ,medicine.anatomical_structure ,Amino Acid Substitution ,chemistry ,Cytoprotection ,Immunology ,Endothelium, Vascular ,Mitogen-Activated Protein Kinases ,Reactive Oxygen Species ,Cardiology and Cardiovascular Medicine ,Oxidative stress ,Signal Transduction - Abstract
Background — Reactive oxygen species play a critical role in inducing apoptosis. The small GTPase p21 Ras and the ERK1/2 MAPK have been proposed as key regulators of the signaling cascade triggered by oxidative stress (H 2 O 2 ). Harvey-Ras (Ha-Ras) and Kirsten-Ras (Ki-Ras) isoforms are so far functionally indistinguishable, because they activate the same downstream effectors, including ERK1/2. Moreover, ERK1/2 signaling has been involved in both protection and induction of apoptosis. Methods and Results — Human umbilical vein endothelial cells (HUVECs) were subjected to H 2 O 2 , and apoptosis was detected by fluorescence-activated cell sorting analysis, fluorescence microscopy, and caspase-3 activation. Transfection of Ha- Ras and Ki- Ras genes in HUVECs was performed to evaluate the response to H 2 O 2 . We have found that, whereas Ha- Ras decreases tolerance to oxidative stress, Ki- Ras has a potent antiapoptotic activity. Both effects are mediated by ERK1/2. Tolerance to H 2 O 2 is encoded by a unique stretch of lysines at the COOH terminus of the Ki-Ras, lacking in Ha-Ras, and it is relatively independent of the farnesylated anchor. Inhibition of p21 Ras signaling by farnesylation inhibitors increased the resistance to apoptosis in Ha-Ras–expressing cells. Conclusions — These findings explain the opposite effects of ERK1/2 stimulation on apoptosis found in different cell types and suggest that local activation of ERK1/2 signaling may account for the opposing response to oxidative stress by Ha-Ras or Ki-Ras–expressing cells. Modulation of cell reactivity to oxidative stress by p21 Ras points to the specific and predictive effects of Ras inhibitors in vivo as potential therapeutic drugs in disorders produced by increase of reactive oxygen species inside the cells.
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- 2002
98. Changes in myocardial cytoskeletal intermediate filaments and myocyte contractile dysfunction in dilated cardiomyopathy: an in vivo study in humans
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Giovanni Cuda, S Di Somma, G Caputo, F. De Vivo, M P Di Benedetto, F Ciaramella, G. Cudemo, M Marotta, G Salvatore, O. De Divitiis, DI SOMMA, S., Marotta, Marcello, Salvatore, G., Cudemo, G., Cuda, G., DE VIVO, F., DI BENEDETTO, M. P., Ciaramella, F., Caputo, G., DE DIVITIIS, O., Di Somma, S., Marotta, M., DI BENEDETTO, M., Ciaramella, S., Caputo, Giuseppe, and DE DIVITIIS, O. .
- Subjects
Adult ,Cardiomyopathy, Dilated ,Male ,medicine.medical_specialty ,Pathology ,Biopsy ,Cardiomyopathy ,desmin ,Vimentin ,Radionuclide ventriculography ,macromolecular substances ,In Vitro Techniques ,Myosins ,Immunoenzyme Techniques ,cardiac biopsy ,vimentin ,Intermediate Filament Proteins ,Internal medicine ,dilated cardiomyopathy, desmin, vimentin, cardiac biopsy, actin-myosin ,actin-myosin ,Humans ,Medicine ,Intermediate filament ,Cytoskeleton ,Aged ,Ejection fraction ,biology ,business.industry ,Myocardium ,Dilated cardiomyopathy ,Middle Aged ,medicine.disease ,Actins ,dilated cardiomyopathy ,Cytoskeletal Proteins ,Basic Research ,Heart failure ,biology.protein ,Cardiology ,Female ,Desmin ,Cardiology and Cardiovascular Medicine ,business - Abstract
AIM—To investigate in vivo the intermediate cytoskeletal filaments desmin and vimentin in myocardial tissues from patients with dilated cardiomyopathy, and to determine whether alterations in these proteins are associated with impaired contractility. METHODS—Endomyocardial biopsies were performed in 12 patients with dilated cardiomyopathy and in 12 controls (six women with breast cancer before anthracycline chemotherapy and six male donors for heart transplantation). Biopsy specimens were analysed by light microscopy and immunochemistry (desmin, vimentin). Myocyte contractile protein function was evaluated by the actin-myosin in vitro motility assay. Left ventricular ejection fraction was assessed by echocardiography and radionuclide ventriculography. RESULTS—Patients with dilated cardiomyopathy had a greater cardiomyocyte diameter than controls (p
- Published
- 2000
99. Co-existence of frataxin and cardiac troponin T gene mutations in a child with Friedreich Ataxia and familial hypertrophic cardiomyopathy
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Giovanni Cuda, Pietro Strisciuglio, Andrea Mussari, Francesco Costanzo, Daniela Concolino, Cuda, G., Mussari, A., Concolino, D., Costanzo, F. S., and Strisciuglio, Pietro
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Male ,Ataxia ,TNNT2 ,DNA Mutational Analysis ,Cardiomyopathy ,Biology ,Gene mutation ,Osteochondrodysplasias ,Sudden death ,Diagnosis, Differential ,Electrocardiography ,Troponin T ,Iron-Binding Proteins ,Genetics ,medicine ,Humans ,Allele ,Genetics (clinical) ,Hypertrophic cardiomyopathy ,Cardiomyopathy, Hypertrophic ,medicine.disease ,Pedigree ,Phosphotransferases (Alcohol Group Acceptor) ,Echocardiography ,Friedreich Ataxia ,Child, Preschool ,Mutation ,Frataxin ,biology.protein ,Female ,medicine.symptom - Abstract
Friedreich Ataxia (FA) is a neurodegenerative disorder characterised by progressive gait disturbance, dysarthria, dysmetria and other coordination disorders. The genetic defect is represented by an expansion of GAA repeats in the frataxin gene (FRDA or X25). Hypertrophic cardiomyopathy is a common finding in FA, and it is widely recognised as specific for the diagnosis of disease status. In this study, we report the co-existence, in a 5-year old boy with FA, of a double mutation in two distinct genes [X25 (A allele: 850 triplets; B allele: 1000 triplets), and cardiac troponin T (TNNT2) (287G>A)]. TNNT2 gene mutations have been previously identified in individuals with a familial form of hypertrophic cardiomyopathy (FHC), an autosomal dominant inherited disease characterised by unexplained cardiac hypertrophy and high incidence of sudden death. Although we cannot rule out the impact of each gene defect on cardiac morphology, it is of interest that these two mechanisms may be acting in a synergistic fashion to produce the extreme degree of cardiac hypertrophy detected in the child. This is, to our knowledge, the first description of a double gene defect in individuals with FA and FHC.
- Published
- 2002
100. Chemotherapy-induced cardiotoxicity: An animal model
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Giovanni Cuda, F. Fortunato, Alfredo Marinelli, Gaetano Salvatore, F. Russo, Luigi Elio Adinolfi, Roberto Ciarcia, S. De Placido, C. Romano, Ar Bianco, C., Romano, Marinelli, Alfredo, F., Fortunato, L., Adinolfi, G., Salvatore, Ciarcia, Roberto, F., Russo, G., Cuda, S., De Placido, A. R., Bianco, Romano, C, Fortunato, F, Adinolfi, L, Salvatore, G, Russo, F, Cuda, G, DE PLACIDO, Sabino, and Bianco, A. R.
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Cancer Research ,Cardiotoxicity ,Chemotherapy ,Cyclophosphamide ,business.industry ,medicine.medical_treatment ,Intraperitoneal injection ,Animal model ,Blood pressure ,Oncology ,Chemotherapy induced ,Anesthesia ,medicine ,business ,medicine.drug ,Epirubicin - Abstract
9673 Background:Cardiac side effects of several antineoplastic drugs are dose-limiting and dose-dependent, thus they are a limitation to the optimal delivery of dose-intensive chemotherapy. Aim of present study was to construct a model to evaluate cardiac damage after chemotherapy. Methods: 45 male Sprague-Dawley rats, about 7 weeks old and weighing 200–225 g were divided into two groups: one group of 30 rats, each animal receiving 5Fluoruracil, 20 mg/kg/wk by intraperitoneal injection, Epirubicin, 3 mg/kg/wk by intravenous injection, Cyclophosphamide, 20 mg/kg/wk by intraperitoneal injection once weekly for 3 consecutive weeks (FEC group) and a second group of 15 rats receiving no drugs (Control group). Observation for mortality and clinical signs was carried out daily, body weight and blood pressure were recorded basally, before each drug administration and weekly until sacrifice. 15 animals, 10 in the FEC group and 5 in the control group, were sacrificed at weeks 3, 5 and 6 after the end of treatment. ...
- Published
- 2004
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