Latest PUBLICATIONS

  • Structure and stability insights into tumour suppressor p53 evolutionary related proteins.
    Publication Date: 04/10/2013, on PloS one
    by Pagano B, Jama A, Martinez P, Akanho E, Bui TT, Drake AF, Fraternali F, Nikolova PV
    DOI: 10.1371/journal.pone.0076014

    The p53 family of genes and their protein products, namely, p53, p63 and p73, have over one billion years of evolutionary history. Advances in computational biology and genomics are enabling studies of the complexities of the molecular evolution of p53 protein family to decipher the underpinnings of key biological conditions spanning from cancer through to various metabolic and developmental disorders and facilitate the design of personalised medicines. However, a complete understanding of the inherent nature of the thermodynamic and structural stability of the p53 protein family is still lacking. This is due, to a degree, to the lack of comprehensive structural information for a large number of homologous proteins and to an incomplete knowledge of the intrinsic factors responsible for their stability and how these might influence function. Here we investigate the thermal stability, secondary structure and folding properties of the DNA-binding domains (DBDs) of a range of proteins from the p53 family using biophysical methods. While the N- and the C-terminal domains of the p53 family show sequence diversity and are normally targets for post-translational modifications and alternative splicing, the central DBD is highly conserved. Together with data obtained from Molecular Dynamics simulations in solution and with structure based homology modelling, our results provide further insights into the molecular properties of evolutionary related p53 proteins. We identify some marked structural differences within the p53 family, which could account for the divergence in biological functions as well as the subtleties manifested in the oligomerization properties of this family.

  • On the differential nature of induced and incidental echolalia in autism.
    Publication Date: 01/10/2013, on Journal of intellectual disability research : JIDR
    by Grossi D, Marcone R, Cinquegrana T, Gallucci M
    DOI: 10.1111/j.1365-2788.2012.01579.x

    Echolalia is a verbal disorder, defined as 'a meaningless repetition of the words of others'. It is pathological, automatic and non-intentional behaviour, often observed in a variety of neurological and psychiatric disorders and above all in autism. We assume that echolalia is an imitative behaviour that is due to difficulties in inhibiting automatic repetition as seen in patients with frontal lobe damage. Our aim is to study the occurrence of echolalia under experimental conditions to investigate the nature of the phenomenon and its relationship with the severity of autism.

  • A new anti-infective strategy to reduce the spreading of antibiotic resistance by the action on adhesion-mediated virulence factors in Staphylococcus aureus.
    Publication Date: 01/10/2013, on Microbial pathogenesis
    by Papa R, Artini M, Cellini A, Tilotta M, Galano E, Pucci P, Amoresano A, Selan L
    DOI: 10.1016/j.micpath.2013.05.003

    Staphylococcus aureus is a flexible microbial pathogen frequently isolated from community-acquired and nosocomial infections. S. aureus expresses a wide array of secreted and cell surface-associated virulence factors, including proteins that promote adhesion to damaged tissue and to the surface of host cells, and that bind proteins in blood to help evade immune responses. Furthermore, surface proteins have a fundamental role in virulence related properties of S. aureus, including biofilm formation. The present study evaluates the anti-infective capabilities of a secreted protein of Serratia marcescens (serratiopeptidase, SPEP), in impairing some staphylococcal virulence-related properties, such as attachment to inert surfaces and adhesion/invasion on eukaryotic cells. SPEP seems to exert its action by modulating specific proteins. It is not assessed if this action is due to the proteolytic activity of SPEP or to a specific mechanism which triggers an out/inside signal. Proteomic studies performed on surface proteins extracted from SPEP treated S. aureus cultures revealed that a number of proteins are affected by the treatment. Among these we found the adhesin/autolysin Atl, SdrD, Sbi, EF-Tu and EF-G. EF-Tu and EF-G are known to perform a variety of function, depending on their cytoplasmic or surface localization. All these factors can facilitate bacterial colonization, persistence and invasion of host tissues. Our results suggest that SPEP could be developed as a potential "anti-infective agent" capable to hinder the entry of S. aureus into human tissues, and also impairs the ability of this pathogen to adhere to prostheses, catheters and medical devices.

  • Predictors of recovery of responsiveness in prolonged anoxic vegetative state. Author reply.
    Publication Date: 01/10/2013, on Neurology
    by Estraneo A, Moretta P, Terme T, Trojano L
    DOI:

  • Disease activity in Graves' ophthalmopathy: diagnosis with orbital MR imaging and correlation with clinical score.
    Publication Date: 01/10/2013, on The neuroradiology journal
    by Tortora F, Cirillo M, Ferrara M, Belfiore MP, Carella C, Caranci F, Cirillo S
    DOI: 10.1177/197140091302600509

    In Graves' ophthalmopathy (GO) it is important to distinguish acute inflammation at an early stage, responsive to immunosuppressive treatment, from inactive fibrotic end stage disease, unresponsive to the same treatment. The purpose of this study was to identify the most relevant signal intensities on orbital MR imaging with contrast administration both to classify patients according to their clinical activity score (defined by a cut-off value of 3) and to make a prediction of patient's CAS. Such threshold was considered as widely used in literature. Sixteen consecutive patients with a diagnosis of GO in different phases of thyroid disease based on clinical and orbital MR imaging signs, and six normal volunteers were examined. Orbital MR imaging was performed on a 1.5 Tesla MR Unit. MR scans were assessed by an experienced neuroradiologist, blinded to the clinical examinations. We found a statistical correlation between CAS and both STIR and contrast enhanced T1-weighted sequences. There was also a statistically significant correlation between STIR and contrast-enhanced T1 images disclosing the possibility of avoiding the injection of contrast medium. Our study proved that signal intensity values on STIR sequence increase in the inflammatory oedematous phase of disease. We confirmed the correlation between signal intensities on this sequence and CAS, showing an increase in signal intensity proportional to the CAS value. So we validated MRI use to establish the activity phase of disease more sensitively than CAS alone.

  • Preamplification procedure for the analysis of ancient DNA samples.
    Publication Date: 25/09/2013, on TheScientificWorldJournal
    by Del Gaudio S, Cirillo A, Di Bernardo G, Galderisi U, Thanassoulas T, Pitsios T, Cipollaro M
    DOI: 10.1155/2013/734676

    In ancient DNA studies the low amount of endogenous DNA represents a limiting factor that often hampers the result achievement. In this study we extracted the DNA from nine human skeletal remains of different ages found in the Byzantine cemetery of Abdera Halkidiki and in the medieval cemetery of St. Spiridion in Rhodes (Greece). Real-time quantitative polymerase chain reaction (qPCR) was used to detect in the extracts the presence of PCR inhibitors and to estimate the DNA content. As mitochondrial DNA was detected in all samples, amplification of nuclear targets, as amelogenin and the polymorphism M470V of the transmembrane conductance regulator gene, yielded positive results in one case only. In an effort to improve amplification success, we applied, for the first time in ancient DNA, a preamplification strategy based on TaqMan PreAmp Master Mix. A comparison between results obtained from nonpreamplified and preamplified samples is reported. Our data, even if preliminary, show that the TaqMan PreAmp procedure may improve the sensitivity of qPCR analysis.

  • New-onset refractory status epilepticus mimicking herpes virus encephalitis.
    Publication Date: 19/09/2013, on Case reports in neurology
    by Puoti G, Elefante A, Saracino D, Capasso A, Cotrufo R, Anello CB
    DOI: 10.1159/000355273

    New-onset refractory status epilepticus (NORSE) is a recently defined clinical entity that describes patients who present with status epilepticus of unclear etiology that is highly refractory to therapy. Magnetic resonance imaging (MRI) of NORSE usually discloses no specific abnormalities except for an occasional mild T2/FLAIR hyperintense signal of the mesial temporal lobe. Here, we report a peculiar case of NORSE in which brain MRI showed massive alteration of both temporal lobes, with features strongly supporting the diagnosis of herpes virus encephalitis, but lacking any laboratory evidence of viral infection in the blood and cerebrospinal fluid. It showed also striking signal alterations in the thalamus, which got worse in the course of the disease. This report emphasizes the possibility that seizure activity alone plays a critical role in both determining the disease and whether it will be self-sustaining.

  • Use of NMR in profiling of cocaine seizures.
    Publication Date: 10/09/2013, on Forensic science international
    by Pagano B, Lauri I, De Tito S, Persico G, Chini MG, Malmendal A, Novellino E, Randazzo A
    DOI: 10.1016/j.forsciint.2013.04.028

    Cocaine is the most widely used illicit drug, and its origin is always the focus of intense investigation aimed at identifying the trafficking routes. Since NMR represents a unique methodology for performing chemical identification and quantification, here it is proposed a strategy based on (1)H NMR spectral analysis in conjunction with multivariate analysis to identify the chemical "fingerprint" of cocaine samples, and to link cocaine samples based on this information. The most relevant spectral regions containing the fingerprint have been identified: δH 0.86-0.96, 1.50-1.56, 5.90-5.93, 6.48-6.52, 7.31-7.34, 7.61-7.63, 7.68-7.72 ppm. The strategy has been applied on samples seized in different times and places in Naples (Italy). The chemical "fingerprint" depend on what plant they were extracted from, where it was cultivated, and which procedures were used for extraction and purification, thus adding significant information in the process toward identification of the trafficking routes for this drug.

  • Inhibition of cardiomyocytes late INa with ranolazine to prevent anthracyclines cardiotoxicity in experimental models in vitro and in vivo.
    Publication Date: 10/09/2013, on Journal of clinical oncology : official journal of the American Society of Clinical Oncology
    by Coppola C, Piscopo G, Cipresso C, Rea D, Tocchetti CG, De Laurentiis M, Arra C, Iaffaioli RV
    DOI: 10.1200/jco.2013.31.26_suppl.170

    170 Background: Anthracyclines are first line drugs against cancer, but produce a well-known cardiomyopathy through multiple mechanisms, which also include Ca2+ overload due to reduced SERCA2a activity and inappropriate opening of the RyR2, and impaired myocardial energetics. Anthracyclines generate Reactive Oxigen and Nitrogen Species, posing the heart at increased demand for oxygen, thus setting the stage for a metabolic ischemia that also activates late INa, the target of ranolazine (RAN). Here, we aim at assessing whether RAN, diminishing intracellular Ca2+ through its inhibition of late INa, and enhancing myocardial glucose utilization (and/or reverting impairment of glucose utilization caused by chemotherapy) blunts anthracyclines cardiotoxicity.

  • Utility of 2D-speckle tracking echocardiography in diagnosis of left ventricular dysfunction in anti-ErbB2 therapy.
    Publication Date: 10/09/2013, on Journal of clinical oncology : official journal of the American Society of Clinical Oncology
    by Piscopo G, Cipresso C, Rea D, Esposito E, Capasso I, Tocchetti CG, Arra C, Coppola C
    DOI: 10.1200/jco.2013.31.26_suppl.169

    169 Background: ErbB2 is overexpressed in about 25% of breast cancers; in the heart, it modulates myocardial development and function. Trastuzumab (T), an anti-ErbB2 inhibitor, has improved the prognosis of patients with breast cancer, but is related to an increased risk of asymptomatic left ventricular (LV) dysfunction (3-34%) and heart failure (2-4%). Conventional measures of ventricular function, such as fractional shortening (FS) and ejection fraction (FE) are insensitive in detecting early cardiomyopathy induced by antineoplastic therapy. Here, we aim at assessing whether myocardial strain by 2D-speckle tracking (ST) is able to identify early LV dysfunction in mice treated with doxorubicin (D) and T, alone or in combination (D+T) and to relate data of cardiac function with tissue alterations.

  • Vesicular and non-vesicular transport feed distinct glycosylation pathways in the Golgi.
    Publication Date: 05/09/2013, on Nature
    by D'Angelo G, Uemura T, Chuang CC, Polishchuk E, Santoro M, Ohvo-Rekilä H, Sato T, Di Tullio G, Varriale A, D'Auria S, Daniele T, Capuani F, Johannes L, Mattjus P, Monti M, Pucci P, Williams RL, Burke JE, Platt FM, Harada A, De Matteis MA
    DOI: 10.1038/nature12423

    Newly synthesized proteins and lipids are transported across the Golgi complex via different mechanisms whose respective roles are not completely clear. We previously identified a non-vesicular intra-Golgi transport pathway for glucosylceramide (GlcCer)--the common precursor of the different series of glycosphingolipids-that is operated by the cytosolic GlcCer-transfer protein FAPP2 (also known as PLEKHA8) (ref. 1). However, the molecular determinants of the FAPP2-mediated transfer of GlcCer from the cis-Golgi to the trans-Golgi network, as well as the physiological relevance of maintaining two parallel transport pathways of GlcCer--vesicular and non-vesicular--through the Golgi, remain poorly defined. Here, using mouse and cell models, we clarify the molecular mechanisms underlying the intra-Golgi vectorial transfer of GlcCer by FAPP2 and show that GlcCer is channelled by vesicular and non-vesicular transport to two topologically distinct glycosylation tracks in the Golgi cisternae and the trans-Golgi network, respectively. Our results indicate that the transport modality across the Golgi complex is a key determinant for the glycosylation pattern of a cargo and establish a new paradigm for the branching of the glycosphingolipid synthetic pathway.

  • Determination of five bisphenols in commercial milk samples by liquid chromatography coupled to fluorescence detection.
    Publication Date: 01/09/2013, on Journal of food protection
    by Grumetto L, Gennari O, Montesano D, Ferracane R, Ritieni A, Albrizio S, Barbato F
    DOI: 10.4315/0362-028X.JFP-13-054

    The presence of five bisphenols, i.e., bisphenol F, bisphenol A, bisphenol B, bisphenol F diglycidyl ether, and bisphenol A diglycidyl ether, was monitored in commercial milk packed in plastic bottles marketed in Italy. The new validated method includes a solid-phase extraction procedure followed by liquid chromatography with fluorescence detection. All positive results were confirmed by liquid chromatography-tandem mass spectrometry analysis. The limits of detection and quantification and the recovery percentages indicated that the method is suitable for detecting bisphenols in milk at concentrations far below the legal limits. Of 68 commercial milk samples analyzed, no bisphenol was found in 27 samples (39.7%), and 41 samples (60.3%) contained one or more bisphenols. The bisphenol most frequently found was bisphenol F (36 samples, 52.9%) followed by bisphenol A (20 samples, 29.4%) and bisphenol B (6 samples, 8.8%). Taking into consideration the limits of detection, no sample contained either bisphenol F diglycidyl ether or bisphenol A diglycidyl ether.

  • Endovascular treatment of CCSVI in patients with multiple sclerosis: clinical outcome of 462 cases.
    Publication Date: 01/09/2013, on Neurological sciences : official journal of the Italian Neurological Society and of the Italian Society of Clinical Neurophysiology
    by Ghezzi A, Annovazzi P, Cocco E, Coarelli G, Lugaresi A, Rovaris M, Patti F, Capello E, Rodegher ME, Moiola L, Malucchi S, Salemi G, De Rossi N, Provinciali L, Perini P, Bergamaschi R, Scarpini E, Lus G, Gallo A, Tola MR, Amato MP, Rottoli MR, Bianchi A, Comi G,
    DOI: 10.1007/s10072-013-1300-5

    Although it is still debated whether chronic cerebro-spinal venous insufficiency (CCSVI) plays a role in multiple sclerosis (MS) development, many patients underwent endovascular treatment (ET) of CCSVI. The objective of the study is to evaluate the outcome and safety of ET in Italian MS patients. Italian MS centers that are part of the Italian MS Study Group were all invited to participate to this retrospective study. A structured questionnaire was used to collect detailed clinical data before and after the ET. Data from 462 patients were collected in 33 centers. ET consisted of balloon dilatation (93 % of cases) or stent application. The mean follow-up duration after ET was 31 weeks. Mean EDSS remained unchanged after ET (5.2 vs. 4.9), 144 relapses occurred in 98/462 cases (21 %), mainly in RR-MS patients. Fifteen severe adverse events were recorded in 3.2 % of cases. Given the risk of severe adverse events and the lack of objective beneficial effects, our findings confirm that at present ET should not be recommended to patients with MS.

  • Vitamin D deficiency in myotonic dystrophy type 1.
    Publication Date: 01/09/2013, on Journal of neurology
    by Terracciano C, Rastelli E, Morello M, Celi M, Bucci E, Antonini G, Porzio O, Tarantino U, Zenobi R, Massa R
    DOI: 10.1007/s00415-013-6984-1

    Myotonic dystrophy type 1 (DM1) is a multisystemic disorder affecting, among others, the endocrine system, with derangement of steroid hormones functions. Vitamin D is a steroid recognized for its role in calcium homeostasis. In addition, vitamin D influences muscle metabolism by genomic and non-genomic actions, including stimulation of the insulin-like-growth-factor 1 (IGF1), a major regulator of muscle trophism. To verify the presence of vitamin D deficit in DM1 and its possible consequences, serum 25-hydroxyvitamin D (25(OH)D), calcium, parathormone (PTH), and IGF1 levels were measured in 32 DM1 patients and in 32 age-matched controls. Bone mineral density (BMD) and proximal muscle strength were also measured by DXA and a handheld dynamometer, respectively. In DM1 patients, 25(OH)D levels were reduced compared to controls, and a significant decrease of IGF1 was also found. 25(OH)D levels inversely correlated with CTG expansion size, while IGF1 levels and muscle strength directly correlated with levels of 25(OH)D lower than 20 and 10 ng/ml, respectively. A significantly higher percentage of DM1 patients presented hyperparathyroidism as compared to controls. Calcium levels and BMD were comparable between the two groups. Oral administration of cholecalciferol in 11 DM1 patients with severe vitamin D deficiency induced a normal increase of circulating 25(OH)D, ruling out defects in intestinal absorption or hepatic hydroxylation. DM1 patients show a reduction of circulating 25(OH)D, which correlates with genotype and may influence IGF1 levels and proximal muscle strength. Oral supplementation with vitamin D should be considered in DM1 and might mitigate muscle weakness.

  • Impulsivity and home-cage activity are decreased by lentivirus-mediated silencing of serotonin transporter in the rat hippocampus.
    Publication Date: 26/08/2013, on Neuroscience letters
    by Zoratto F, Tringle AL, Bellenchi G, Speranza L, Travaglini D, di Porzio U, Perrone-Capano C, Laviola G, Dreyer JL, Adriani W
    DOI: 10.1016/j.neulet.2013.05.076

    Brain serotonin (5-HT) systems modulate emotional, motivational and cognitive processes. Mutations in the serotonin transporter (SERT) gene have been associated with susceptibility towards the development of several psychiatric disorders, both in humans and animal models. Present approach exploited a bilateral intra-hippocampus stereotaxic inoculation of lentiviruses, for enduring in vivo silencing of SERT. Control rats were bilaterally inoculated with heat-inactivated lentiviruses. These Lenti-SERT vectors were intended to eventually manipulate the neurotransmitter reuptake at synaptic level, thus enhancing tonic 5-HT transmission. We investigated whether such manipulation could induce behavioural alterations relevant to the modelling of ADHD, in particular symptoms of hyperactivity and impulsivity. Wistar rats were monitored for spontaneous home-cage locomotor activity and studied for impulsivity (Intolerance-to-Delay task). Results show that rats inoculated with Lenti-SERT vectors exhibited less pronounced circadian peaks of activity than controls. Moreover, Lenti-SERT compared to control rats exhibited a transient increase in choice for a delayed-larger reward over an immediate-small reward. This suggests that enhanced hippocampal serotonergic transmission produced a profile of restfulness and a decrease in cognitive impulsivity. This phenotype is consistent with available data both on 5-HT manipulations and hippocampal lesions. In conclusion, present findings may possibly disclose novel avenues towards the development of innovative therapeutical approaches for behavioural symptoms relevant to ADHD.