@article {10798, title = {Imaging and therapy of ovarian cancer: clinical application of nanoparticles and future perspectives.}, journal = {Theranostics}, volume = {8}, year = {2018}, month = {2018}, pages = {4279-4294}, abstract = {

Despite significant advances in cancer diagnostics and treatment, ovarian cancers (OC) continue to kill more than 150,000 women every year worldwide. Due to the relatively asymptomatic nature and the advanced stage of the disease at the time of diagnosis, OC is the most lethal gynecologic malignancy. The current treatment for advanced OC relies on the synergistic effect of combining surgical cytoreduction and chemotherapy; however, beside the fact that chemotherapy resistance is a major challenge in OC management, new imaging strategies are needed to target microscopic lesions and improve both cytoreductive surgery and patient outcomes. In this context, nanostructured probes are emerging as a new class of medical tool that can simultaneously provide imaging contrast, target tumor cells, and carry a wide range of medicines resulting in better diagnosis and therapeutic precision. Herein we summarize several exemplary efforts in nanomedicine for addressing unmet clinical needs.

}, issn = {1838-7640}, doi = {10.7150/thno.26345}, author = {Di Lorenzo, Giovanni and Ricci, Giuseppe and Severini, Giovanni Maria and Romano, Federico and Biffi, Stefania} } @article {10550, title = {Synthesis of Lipophilic Core-Shell FeO@SiO@Au Nanoparticles and Polymeric Entrapment into Nanomicelles: A Novel Nanosystem for in Vivo Active Targeting and Magnetic Resonance-Photoacoustic Dual Imaging.}, journal = {Bioconjug Chem}, volume = {28}, year = {2017}, month = {2017 05 17}, pages = {1382-1390}, abstract = {

In this work, iron/silica/gold core-shell nanoparticles (FeO@SiO@Au NPs) characterized by magnetic and optical properties have been synthesized to obtain a promising theranostic platform. To improve their biocompatibility, the obtained multilayer nanoparticles have been entrapped in polymeric micelles, decorated with folic acid moieties, and tested in vivo for photoacoustic and magnetic resonance imaging detection of ovarian cancer.

}, keywords = {Animals, Cell Proliferation, Female, Ferric Compounds, Folic Acid, Gold, Humans, Image Processing, Computer-Assisted, Magnetic Resonance Imaging, Magnetite Nanoparticles, Mice, Mice, Inbred BALB C, Mice, Nude, Micelles, Multimodal Imaging, Ovarian Neoplasms, Photoacoustic Techniques, Polymers, Silicon Dioxide, Tumor Cells, Cultured, Xenograft Model Antitumor Assays}, issn = {1520-4812}, doi = {10.1021/acs.bioconjchem.7b00076}, author = {Monaco, Ilaria and Arena, Francesca and Biffi, Stefania and Locatelli, Erica and Bortot, Barbara and La Cava, Francesca and Marini, Giada Maria and Severini, Giovanni Maria and Terreno, Enzo and Comes Franchini, Mauro} } @article {8348, title = {Clinical Spectrum of PRKAG2 Syndrome.}, journal = {Circ Arrhythm Electrophysiol}, volume = {9}, year = {2016}, month = {2016 Jan}, pages = {e003121}, keywords = {AMP-Activated Protein Kinases, DNA, Heart Conduction System, Humans, Mutation, Wolff-Parkinson-White Syndrome}, issn = {1941-3084}, doi = {10.1161/CIRCEP.115.003121}, author = {Porto, Andrea Giuseppe and Brun, Francesca and Severini, Giovanni Maria and Losurdo, Pasquale and Fabris, Enrico and Taylor, Matthew R G and Mestroni, Luisa and Sinagra, Gianfranco} } @article {3610, title = {F402L variant in NLRP12 in subjects with undiagnosed periodic fevers and in healthy controls.}, journal = {Clin Exp Rheumatol}, volume = {32}, year = {2014}, month = {2014 Nov-Dec}, pages = {993-4}, keywords = {Cryopyrin-Associated Periodic Syndromes, Female, Humans, Intracellular Signaling Peptides and Proteins, Male, Mutation}, issn = {0392-856X}, author = {De Pieri, Carlo and Vuch, Josef and Athanasakis, Emmanouil and Severini, Giovanni Maria and Crovella, Sergio and Bianco, Anna Monica and Tommasini, Alberto} } @article {1799, title = {Β-hexosaminidase over-expression affects lysosomal glycohydrolases expression and glycosphingolipid metabolism in mammalian cells.}, journal = {Mol Cell Biochem}, volume = {363}, year = {2012}, month = {2012 Apr}, pages = {109-18}, abstract = {

Lysosomes are not only degrading organelles but also involved in other critical cellular processes. In addition, active lysosomal glycohydrolases have been detected in an extra-lysosomal compartment: the presence of glycohydrolases on the plasma membrane (PM) has been widely demonstrated, and a possible role on the modification of the cell surface glycosphingolipids (GSL) participating in the modulation of cell functions such as cell-to-cell interactions and signal transduction pathways has been proposed. On this basis, the coordinated expression of lysosomal glycohydrolases and their translocation to the PM appear to be crucial for many cellular events. In this paper, we report evidence for the existence of a coordinated mechanism regulating the expression/activity of both lysosomal and PM-associated glycohydrolases. We show that the over-expression of the acidic glycohydrolase β-hexosaminidase α-subunit in mouse NIH/3T3 fibroblasts induces the increased expression of the Hex β-subunit necessary to form the active isoenzyme dimers as well as of other glycohydrolases participating in the GSL catabolism, such as β-galactosidase and β-glucocerebrosidase. More interestingly, this regulatory effect was also extended to the PM-associated hydrolases. In addition, transfected cells displayed a rearrangement of the GSL expression pattern that cannot be simply explained by the increased activity of a single enzyme. These observations clearly indicate that the expression level of metabolically related glycohydrolases is regulated in a coordinated manner and this regulation mechanism also involves the PM-associated isoforms.

}, keywords = {Animals, beta-Hexosaminidase alpha Chain, Cell Membrane, Exocytosis, Fibroblasts, Glycoside Hydrolases, Glycosphingolipids, Humans, Lysosomes, Mice, NIH 3T3 Cells, Transfection}, issn = {1573-4919}, doi = {10.1007/s11010-011-1163-0}, author = {Tancini, Brunella and Magini, Alessandro and Bortot, Barbara and Polchi, Alice and Urbanelli, Lorena and Sonnino, Sandro and Severini, Giovanni Maria and Emiliani, Carla} } @article {1779, title = {High-throughput genotyping robot-assisted method for mutation detection in patients with hypertrophic cardiomyopathy.}, journal = {Diagn Mol Pathol}, volume = {20}, year = {2011}, month = {2011 Sep}, pages = {175-9}, abstract = {

Hypertrophic cardiomyopathy (HCM) is the most frequent autosomal dominant genetic heart muscle disease and the most common cause of sudden cardiac death in young people (under 30 y of age), who are often unaware of their underlying condition. Genetic screening is now considered a fundamental tool for clinical management of HCM families. However, the high genetic heterogeneity of HCM makes genetic screening very expensive. Here, we propose a new high-throughput genotyping method based on a HCM 96-well sequencing plate for the analysis of 8 of the most frequent HCM-causing sarcomeric genes by automating several processes required for direct sequencing, using a commercially available robotic systems and routinely used instruments. To assess the efficiency of the robot-assisted method, we have analyzed the entire coding sequence and flanking intronic sequences of the 8 sarcomeric genes in samples from 18 patients affected by HCM and their relatives, which revealed 9 different mutations, 3 of which were novel. The automated, robot-assisted assembling of polymerase chain reaction, purification of polymerase chain reaction products, and assembly of sequencing reactions resulted in a substantial saving of time, reagent costs, and reduction of human errors, and can therefore be proposed as a primary strategy for mutation identification in HCM genetic screening in many medical genetic laboratories.

}, keywords = {Cardiomyopathy, Hypertrophic, DNA Mutational Analysis, Genetic Predisposition to Disease, Genetic Testing, Genotyping Techniques, High-Throughput Nucleotide Sequencing, Humans, Muscle Proteins, Mutation, Robotics}, issn = {1533-4066}, doi = {10.1097/PDM.0b013e31820b34fb}, author = {Bortot, Barbara and Athanasakis, Emmanouil and Brun, Francesca and Rizzotti, Diego and Mestroni, Luisa and Sinagra, Gianfranco and Severini, Giovanni Maria} } @article {1722, title = {Quantification of heteroplasmic mitochondrial DNA mutations for DNA samples in the low picogram range by nested real-time ARMS-qPCR.}, journal = {Diagn Mol Pathol}, volume = {20}, year = {2011}, month = {2011 Jun}, pages = {117-22}, abstract = {

In many mitochondrial diseases, different clinical manifestations are related to tissue-specific distribution of mutated mitochondrial DNA (mtDNA). In this study, we describe an assay for the determination of mutated mtDNA copy number in small clinical samples, using standard polymerase chain reaction (PCR) followed by SYBR Green real-time allelic-specific PCR [amplification refractory mutation system-quantitative PCR (ARMS-qPCR)]. To assess the degree of heteroplasmy in a patient harboring 2 cosegregating mtDNA mutations (4415A>G and 9922A>C) starting from picogram amounts of DNA, we first amplified the mutated target sequence by standard PCR, and then analyzed it by real-time ARMS-qPCR. To validate this method, we analyzed by real-time ARMS-qPCR the PCR amplification products derived from different mixtures containing known proportions of mutant and wild-type cloned mtDNA fragments. The correlation coefficient of 0.994 between expected and observed values for the percentage of mutant A4415G confirms that the relative proportion of mutated and wild-type mtDNA was maintained after the first PCR amplification. This method allows the precise quantification of heteroplasmic mutations in DNA samples extracted from hairs, urine, small stomach biopsies, and, more importantly, single-muscle fiber, with a limit of detection close to 0.5\%. This nested real-time ARMS-PCR represents a rapid, efficient, and less expensive method for the detection and quantification of heteroplasmic mutant mtDNA, even in very small clinical samples.

}, keywords = {Child, DNA, Mitochondrial, Humans, Mitochondrial Diseases, Mutation, Polymerase Chain Reaction, Sensitivity and Specificity}, issn = {1533-4066}, doi = {10.1097/PDM.0b013e3181efe2c6}, author = {Biffi, Stefania and Bortot, Barbara and Carrozzi, Marco and Severini, Giovanni Maria} }