• Specific Expression of a New Bruton Tyrosine Kinase Isoform (p65BTK) in the Glioblastoma Gemistocytic Histotype.

    Publication Date: 24/01/2019, on Frontiers in molecular neuroscience
    by Sala L, Cirillo G, Riva G, Romano G, Giussani C, Cialdella A, Todisco A, Virtuoso A, Cerrito MG, Bentivegna A, Grassilli E, Ardizzoia A, Bonoldi E, Giovannoni R, Papa M, Lavitrano M
    DOI: 10.3389/fnmol.2019.00002

    Bruton's tyrosine-kinase (BTK) is a non-receptor tyrosine kinase recently associated with glioma tumorigenesis and a novel prognostic marker for poor survival in patients with glioma. The p65BTK is a novel BTK isoform involved in different pathways of drug resistance of solid tumors, thus we aimed to investigate the expression and the putative role of p65BTK in tumors of the central nervous system (CNS). We selected a large cohort of patients with glial tumors ( = 71) and analyzed the expression of p65BTK in different histotypes and correlation with clinical parameters. Sections were stained with glial fibrillary acidic protein (GFAP), p53, epidermal growth factor receptor (EGFR), S100, vimentin, and epithelial membrane antigen (EMA) antibodies. Glioma stem cell (GSC) lines, isolated from glioblastoma multiforme (GBM), were treated with different concentrations of ibrutinib, a specific inhibitor of BTK, in order to evaluate their metabolic activity, mitotic index and mortality. Moreover, an orthotopic xenotransplant of GSC from human GBM was used to evaluate the expression of p65BTK in the brain of immunodeficient mice. p65BTK was expressed in GSC and in gemistocytes in human gliomas at different histological grade. We found a significant correlation between BTK expression and low-grade (LG) tumors ( ≤ 0.05) and overall survival (OS) of patients with grade III gliomas ( ≤ 0.05), suggestive of worst prognosis. Interestingly, the expression of p65BTK remained restricted exclusively to gemistocytic cells in the xenograft mouse model. Ibrutinib administration significantly reduced metabolic activity and mitotic index and increased mortality in GSC, highlighting the specific role of p65BTK in cell proliferation and survival. In conclusion, our data demonstrated that p65BTK is expressed in glioma tumors, restricted to gemistocytic cells, has a key role in GSC and has a bad prognostic value, thus highlighting the importance of future research for targeted therapy of human gliomas.

  • Encapsulation of the dinuclear trithiolato-bridged arene ruthenium complex diruthenium-1 in an apoferritin nanocage: structure and cytotoxicity.

    Publication Date: 23/01/2019, on ChemMedChem
    by Petruk G, Monti DM, Ferraro G, Pica A, D'Elia L, Pane F, Amoresano A, Furrer J, Kowalski K, Merlino A
    DOI: 10.1002/cmdc.201800805

    The effects of the encapsulation of the cytotoxic dinuclear trithiolato-bridged arene Ru complex [(η6-p-MeC6H4Pri)2Ru2(μ2-S-p-C6H4But)3]Cl (DiRu-1) within the apoferritin (AFt) nanocage were investigated. The DiRu-1-AFt nanocarrier was characterized by UV-Vis spectroscopy, ICP MS, CD and X-ray crystallography. In contrast to previously reported Au- and Pt- based drug-loaded AFt carriers, no direct interactions between DiRu-1 and AFt were evidenced. DiRu-1-AFt is cytotoxic towards immortalized murine fibroblast BALB/c-3T3 transformed with SV40 virus (SVT2) and human epidermoid carcinoma A431 malignant cells and exhibits a moderate selectivity for these cancer cells over the normal BALB/c-3T3 cells. DiRu-1-AFt triggers ROS production, depolarization of mitochondrial membrane potential and induces cell death via p53-mediated apoptosis. The comparison between our data and previous results suggest that the existence of specific interactions between a metal-based drug and AFt within the protein cage is not essential for drug encapsulation.

  • Neutralization of interleukin-17 rescues amyloid-β-induced neuroinflammation and memory impairment.

    Publication Date: 23/01/2019, on British journal of pharmacology
    by Cristiano C, Volpicelli F, Lippiello P, Buono B, Raucci F, Piccolo M, Iqbal AJ, Irace C, Miniaci MC, Perrone Capano C, Calignano A, Mascolo N, Maione F
    DOI: 10.1111/bph.14586

    Alzheimer's disease (AD) is a common neurodegenerative disease characterized by a neuroinflammatory state and to date, there is no cure and its treatment represents a large unmet clinical need. The involvement of T helper 17 cells in the pathogenesis of AD-related neuroinflammation has been reported in several studies, however the role of the main cytokine, IL-17, has not been well addressed. Herein, we investigate the effects of IL-17 neutralizing antibody (IL-17Ab) injected by intracerebroventricular (ICV) or intranasal (IN) routes on amyloid-β-induced neuroinflammation and memory impairment in mice.

  • Antioxidant and Anti-Inflammatory Activities of Buffalo Milk δ-Valerobetaine.

    Publication Date: 20/01/2019, on Journal of agricultural and food chemistry
    by D'Onofrio N, Balestrieri A, Neglia G, Monaco A, Tatullo M, Casale R, Limone A, Balestrieri ML, Campanile G
    DOI: 10.1021/acs.jafc.8b07166

    δ-Valerobetaine (δVB), a constitutive metabolite of ruminant milk, is produced in the rumen from free dietary Nε-trimethyllysine occurring ubiquitously in vegetable kingdom. Biological role of δVB is poorly known. Here, the antioxidant and anti-inflammatory potential of buffalo milk δVB was tested in vitro during high-glucose (HG)-induced endothelial damage. Results indicated that δVB (0.5 mM) ameliorated the HG cytotoxicity (0.57±0.02 vs. 0.41±0.018 O.D. (P<0.01). Buffalo milk extracts enriched with δVB showed significant efficacy in decreasing reactive oxygen species, lipid peroxidation, and cytokine release during HG treatment compared to milk extracts alone (P<0.05). δVB reduced the HG-activated inflammatory signal by modulating SIRT1 (0.96±0.05 vs. 0.85±0.04 AU), SIRT6 (0.82±0.04 vs. 0.61±0.03 AU), and NF-κB (0.85±0.03 vs. 1.23±0.03 AU) (P<0.05). On the whole, our data show the first evidence of δVB efficacy in reducing endothelial oxidative stress and inflammation, suggesting a potential role of this betaine as a novel dietary compound with health-promoting properties.

  • Molecular determinants of ER-Golgi contacts identified through a new FRET-FLIM system.

    Publication Date: 18/01/2019, on The Journal of cell biology
    by Venditti R, Rega LR, Masone MC, Santoro M, Polishchuk E, Sarnataro D, Paladino S, D'Auria S, Varriale A, Olkkonen VM, Di Tullio G, Polishchuk R, De Matteis MA
    DOI: 10.1083/jcb.201812020

    ER-TGN contact sites (ERTGoCS) have been visualized by electron microscopy, but their location in the crowded perinuclear area has hampered their analysis via optical microscopy as well as their mechanistic study. To overcome these limits we developed a FRET-based approach and screened several candidates to search for molecular determinants of the ERTGoCS. These included the ER membrane proteins VAPA and VAPB and lipid transfer proteins possessing dual (ER and TGN) targeting motifs that have been hypothesized to contribute to the maintenance of ERTGoCS, such as the ceramide transfer protein CERT and several members of the oxysterol binding proteins. We found that VAP proteins, OSBP1, ORP9, and ORP10 are required, with OSBP1 playing a redundant role with ORP9, which does not involve its lipid transfer activity, and ORP10 being required due to its ability to transfer phosphatidylserine to the TGN. Our results indicate that both structural tethers and a proper lipid composition are needed for ERTGoCS integrity.

  • Ultra-short term HRV features as surrogates of short term HRV: a case study on mental stress detection in real life.

    Publication Date: 17/01/2019, on BMC medical informatics and decision making
    by Castaldo R, Montesinos L, Melillo P, James C, Pecchia L
    DOI: 10.1186/s12911-019-0742-y

    This paper suggests a method to assess the extent to which ultra-short Heart Rate Variability (HRV) features (less than 5 min) can be considered as valid surrogates of short HRV features (nominally 5 min). Short term HRV analysis has been widely investigated for mental stress assessment, whereas the validity of ultra-short HRV features remains unclear. Therefore, this study proposes a method to explore the extent to which HRV excerpts can be shortened without losing their ability to automatically detect mental stress.

  • Milk From Cow Fed With High Forage/Concentrate Ratio Diet: Beneficial Effect on Rat Skeletal Muscle Inflammatory State and Oxidative Stress Through Modulation of Mitochondrial Functions and AMPK Activity.

    Publication Date: 17/01/2019, on Frontiers in physiology
    by Trinchese G, Cavaliere G, Penna E, De Filippo C, Cimmino F, Catapano A, Musco N, Tudisco R, Lombardi P, Infascelli F, Messina G, Muredda L, Banni S, Monda M, Crispino M, Mollica MP
    DOI: 10.3389/fphys.2018.01969

    Milk and dairy products are relevant components of daily diet and are part of dietary recommendation in many countries due to their content of key nutrients. However, the relatively high content of saturated fat of the milk and its extensive usage for every age group raises concerns about its potential negative health effects. Therefore, in the last years, several researchers dedicated their attention to milk production and quality. Milk fatty acids profile depend on cow feeding and in particular on the type of forage and concentrate and forage/concentrate ratio. It was demonstrated that feeding dairy cows with a 70/30 forage/concentrate ratio yields milk with a low ω6:ω3 ratio and high CLA levels. In this work, we demonstrated that the supplementation of rats diet with this high forage milk (HFM) results, in the skeletal muscle of these animals, in a reduced lipid content and inflammation levels, and an improved mitochondrial lipid oxidation, and redox status through modulation of AMPK activity.

  • Crowding and conformation interplay on human DNA G-quadruplex by ultraviolet resonant Raman scattering.

    Publication Date: 14/01/2019, on Physical chemistry chemical physics : PCCP
    by Di Fonzo S, Bottari C, Brady JW, Tavagnacco L, Caterino M, Petraccone L, Amato J, Giancola C, Cesàro A
    DOI: 10.1039/c8cp04728f

    The G-quadruplex-forming telomeric sequence (TTAGGG)4TT was investigated by polarized Ultraviolet Resonance Raman Scattering (UVRR) at 266 nm. The presence of 40% poly(ethylene glycol) and the so-called "self-crowding" condition were used to induce the hybrid-to-parallel topology transition. Analysis of frequency shifts with temperature showed the role of several functional groups in the topological transitions and provides structural dynamical information. Circular dichroism under similar conditions was used as a reference. UVRR shed light on the effect of intramolecular interactions and of local and environmental dynamics in promoting different G-quadruplex topologies, induced by solution conditions or by temperature changes. Overall, these findings showed the enormous potential of this spectroscopy for G-quadruplex conformational studies.

  • Comparison of alternate and original forms of the Montreal Cognitive Assessment (MoCA): an Italian normative study.

    Publication Date: 14/01/2019, on Neurological sciences : official journal of the Italian Neurological Society and of the Italian Society of Clinical Neurophysiology
    by Siciliano M, Chiorri C, Passaniti C, Sant'Elia V, Trojano L, Santangelo G
    DOI: 10.1007/s10072-019-3700-7

    The Montreal Cognitive Assessment (MoCA) is a screening test widely used in clinical practice and suited for detection of Mild Cognitive Impairment. Alternate forms of the MoCA were developed to avoid "learning effect" in serial assessments, and the present study aimed at investigating inter-form parallelism and at providing normative values for the Italian versions of MoCAs 2 and 3.

  • Is Shouldice the best NON-MESH inguinal hernia repair technique? A systematic review and network metanalysis of randomized controlled trials comparing Shouldice and Desarda.

    Publication Date: 09/01/2019, on International journal of surgery (London, England)
    by Bracale U, Melillo P, Piaggio D, Pecchia L, Cuccurullo D, Milone M, De Palma GD, Cavallaro G, Campanelli G, Merola G, Stabilini C
    DOI: 10.1016/j.ijsu.2019.01.001

    Current guidelines state that the Shouldice technique has lower recurrence rates than other suture repairs and therefore is strongly recommended in non-mesh inguinal hernia repair. Recently a new tissue repair technique has been proposed by Desarda and studied in trials against Lichtenstein technique.

  • On the pH-Modulated Ru-Based Prodrug Activation Mechanism.

    Publication Date: 07/01/2019, on Inorganic chemistry
    by Caterino M, Herrmann M, Merlino A, Riccardi C, Montesarchio D, Mroginski MA, Musumeci D, Ruffo F, Paduano L, Hildebrandt P, Kozuch J, Vergara A
    DOI: 10.1021/acs.inorgchem.8b02667

    The Ru-based prodrug AziRu efficiently binds to proteins, but the mechanism of its release is still disputed. Herein, in order to test the hypothesis of a reduction-mediated Ru release from proteins, a Raman-assisted crystallographic study on AziRu binding to a model protein (hen egg white lysozyme), in two different oxidation states, Ru and Ru, was carried out. Our results indicate Ru reduction, but the Ru release upon reduction is dependent on the reducing agent. To better understand this process, a pH-dependent, spectroelectrochemical surface-enhanced Raman scattering (SERS) study was performed also on AziRu-functionalized Au electrodes as a surrogate and simplest model system of Ru- and Ru-based drugs. This SERS study provided a p K of 6.0 ± 0.4 for aquated AziRu in the Ru state, which falls in the watershed range of pH values separating most cancer environments from their physiological counterparts. These experiments also indicate a dramatic shift of the redox potential E by >600 mV of aquated AziRu toward more positive potentials upon acidification, suggesting a selective AziRu reduction in cancer lumen but not in healthy ones. It is expected that the nature of the ligands (e.g., pyridine vs imidazole, present in well-known Ru complex NAMI-A) will modulate the p K and E, without affecting the underlying reaction mechanism.

  • A novel SLC20A2 gene mutation causing primary familial brain calcification in an Ukrainian patient.

    Publication Date: 03/01/2019, on Neurological sciences : official journal of the Italian Neurological Society and of the Italian Society of Clinical Neurophysiology
    by Oliva M, Capaldo G, D'Amico A, Colavito D, Elefante A, Straccia G, Ugga L, Puoti G
    DOI: 10.1007/s10072-018-3684-8

  • Methods for Monitoring Macroautophagy in Pancreatic Cancer Cells.

    Publication Date: 01/01/2019, on Methods in molecular biology (Clifton, N.J.)
    by Vidoni C, Ferraresi A, Seca C, Secomandi E, Isidoro C
    DOI: 10.1007/978-1-4939-8879-2_18

    Macroautophagy is a catabolic process through which redundant, aged, or damaged cellular structures are first enclosed within double-membrane vesicles (called autophagosomes), and thereafter degraded within lysosomes. Macroautophagy provides a primary route for the turnover of macromolecules, membranes and organelles, and as such plays a major role in cell homeostasis. As part of the stress response, autophagy is crucial to determine the cell fate in response to extracellular or intracellular injuries. Autophagy is involved in cancerogenesis and in cancer progression. Here we illustrate the essential methods for monitoring autophagy in pancreatic cancer cells.

  • Early posterior vitreous detachment is associated with LAMA5 dominant mutation.

    Publication Date: 27/12/2018, on Ophthalmic genetics
    by Napolitano F, Di Iorio V, Di Iorio G, Melone MAB, Gianfrancesco F, Simonelli F, Esposito T, Testa F, Sampaolo S
    DOI: 10.1080/13816810.2018.1558261

    Extracellular matrix molecular components, previously linked to multisystem syndromes include collagens, fibrillins and laminins. Recently, we described a novel multisystem syndrome caused by the c.9418G>A p.(V3140M) mutation in the laminin alpha-5 (LAMA5) gene, which affects connective tissues of all organs and apparatus in a three generation family. In the same family, we have also reported a myopic trait, which, however, was linked to the Prolyl 4-hydroxylase subunit alpha-2 (P4HA2) gene. Results of investigation on vitreous changes and their pathogenesis are reported in the present study.

  • Correction to: Thrombus aspiration in hyperglycemic ST-elevation myocardial infarction (STEMI) patients: clinical outcomes at 1-year follow-up.

    Publication Date: 27/12/2018, on Cardiovascular diabetology
    by Sardu C, Barbieri M, Balestrieri ML, Siniscalchi M, Paolisso P, Calabrò P, Minicucci F, Signoriello G, Portoghese M, Mone P, D'Andrea D, Gragnano F, Bellis A, Mauro C, Paolisso G, Rizzo MR, Marfella R
    DOI: 10.1186/s12933-018-0804-y

    Following publication of the original article [1], the authors reported an error in Acknowledgment section. The last sentence should read as "All authors have read and approval the submission to Cardiovascular Diabetology.