European Institute of Oncology

6 articles published in JoVE

• Biochemistry

  
  A Mass Spectrometry-Based Proteomics Approach for Global and High-Confidence Protein R-Methylation Analysis Marianna Maniaci*^1,2, Federica Marini*¹, Enrico Massignani^1,2, Tiziana Bonaldi^{1 1}Department of Experimental Oncology, European Institute of Oncology IRCCS (IEO), ²European School of Molecular Medicine (SEMM), c/o Campus IFOM-IEO Protein Arginine (R)-methylation is a wide-spread post-translational modification regulating multiple biological pathways. Mass spectrometry is the best technology to globally profile the R-methyl-proteome, when coupled to biochemical approaches for modified peptide enrichment. The workflow designed for the high confidence identification of global R-methylation in human cells is described here.

• Medicine

  
  A Whole Body Dosimetry Protocol for Peptide-Receptor Radionuclide Therapy (PRRT): 2D Planar Image and Hybrid 2D+3D SPECT/CT Image Methods Maria Luisa Belli¹, Emilio Mezzenga¹, Valentina Di Iorio², Monica Celli³, Paola Caroli³, Elisabeth Canali³, Federica Matteucci³, Elisa Tardelli³, Ilaria Grassi³, Maddalena Sansovini³, Silvia Nicolini³, Stefano Severi³, Marta Cremonesi⁴, Mahila Ferrari⁵, Giovanni Paganelli³, Anna Sarnelli^{1 1}Medical Physics Unit, Istituto Scientifico Romagnolo per lo Studio e la Cura dei Tumori (IRST) IRCCS, Meldola, Italy, ²Oncology Pharmacy, Istituto Scientifico Romagnolo per lo Studio e la Cura dei Tumori (IRST) IRCCS, Meldola, Italy, ³Nuclear Medicine Unit, Istituto Scientifico Romagnolo per lo Studio e la Cura dei Tumori (IRST) IRCCS, Meldola, Italy, ⁴Radiation Research Unit, Istituto Europeo di Oncologia (IEO) IRCCS, Milano, Italy, ⁵Medical Physics Unit, Istituto Europeo di Oncologia (IEO) IRCCS, Milano, Italy This method estimates the absorbed dose of different structures for peptide-receptor-radionuclide-therapy (PRRT) with the possibility of avoiding organ overlap on 2D-projections. Serial whole-body planar images permit estimation of mean absorbed doses along the whole body, while the hybrid approach, combining planar images and 3D-SPECT/CT image, overcomes the limitations of structure overlapping.

• Biology

  
  Oligomerization Dynamics of Cell Surface Receptors in Living Cells by Total Internal Reflection Fluorescence Microscopy Combined with Number and Brightness Analysis Moreno Zamai¹, Antonio Trullo², Elvira Arza¹, Ugo Cavallaro³, Valeria R. Caiolfa^{1,2 1}Unit of Microscopy and Dynamic Imaging, Centro Nacional de Investigaciones Cardiovasculares (CNIC), Madrid, Spain, ²Centro di Imaging Sperimentale, Ospedale San Raffaele, Milan, Italy, ³Unit of Gynecological Oncology Research, European Institute of Oncology IRCCS, Milan, Italy We describe an imaging approach for the determination of the average oligomeric state of mEGFP-tagged-receptor oligomers induced by ligand binding in the plasma membrane of living cells. The protocol is based on Total Internal Reflection Fluorescence (TIRF) microscopy combined with Number and Brightness (N&B) analysis.

• Immunology and Infection

  
  Using Robotic Systems to Process and Embed Colonic Murine Samples for Histological Analyses Fulvia Milena Cribiù*¹, Claudia Burrello*², Roberta Tacchi¹, Francesca Boggio¹, Dario Ricca¹, Flavio Caprioli^3,4, Stefano Ferrero¹, Federica Facciotti^{2 1}Pathology Unit, Fondazione IRCCS Cà Granda, Ospedale Maggiore Policlinico, ²Department of Experimental Oncology, European Institute of Oncology, ³ Lack of standardization for murine tissue processing reduces the quality of murine histopathological analysis as compared to human specimens. Here, we present a protocol to perform histopathological examination of murine inflamed and uninflamed colonic tissues to show the feasibility of robotic systems routinely used for processing and embedding human samples.

• Biology

  
  The ChroP Approach Combines ChIP and Mass Spectrometry to Dissect Locus-specific Proteomic Landscapes of Chromatin Monica Soldi¹, Tiziana Bonaldi^{1 1}Department of Experimental Oncology, European Institute of Oncology By combining native and crosslinking chromatin immunoprecipitation with high-resolution Mass Spectrometry, ChroP approach enables to dissect the composite proteomic architecture of histone modifications, variants and non-histonic proteins synergizing at functionally distinct chromatin domains.

• Biology

  
  A Novel Method for the Culture and Polarized Stimulation of Human Intestinal Mucosa Explants Katerina Tsilingiri¹, Angelica Sonzogni², Flavio Caprioli³, Maria Rescigno^{1 1}Department of Experimental Oncology, European Institute of Oncology, ²Department of Pathology and Laboratory Medicine, European Institute of Oncology, ³U.O. Gastroenterologia 2, IRCCS Ca' Granda, Ospedale Policlinico di Milano We introduce a novel method for the maintenance of human intestinal mucosa in culture and monitoring of the response to various types of stimuli over at least 24 hrs. With our method, the polarity of the tissue is maintained, allowing for a physiological stimulation via the apical route.