Latest PUBLICATIONS

  • Plasma nitroproteome of kidney disease patients.
    Publication Date: 01/02/2011, on Amino acids
    by Piroddi M, Palmese A, Pilolli F, Amoresano A, Pucci P, Ronco C, Galli F
    DOI: 10.1007/s00726-010-0693-1

    3'-Nitrotyrosine (3NT) is a post-translational modification (PTM) of body fluids and tissues that is sustained by chronic inflammation and oxidative stress, two main clinical traits of chronic kidney disease (CKD). Despite this background, protein targets and their differential susceptibility to in vivo nitration remain almost completely unexplored in CKD. This study reports a first investigation of plasma nitroproteome in these patients, carried out by both immunorecognition and LC-MS/MS techniques. Plasma proteins of chronic and end-stage KD patients showed a higher burden of nitration than in healthy controls, but main nitration targets appeared to be the same in these populations. Immunoblotting data showed that uremic albumin is largely represented in the uremic nitroproteome together with fibrinogen chains (A, B and C), transferrin, α1-antitrypsin, complement factor D, haptoglobin, and IgG light and heavy chains. However, immunopurification and affinity chromatography experiments demonstrated that the relative content of 3NT on the albumin molecule was very low when compared with that of the remaining plasma proteins. The uremic nitroproteome was investigated using also plasma proteins obtained by in vivo ultrafiltration from patients treated with protein leaking or standard high-flux hemodialyzers. The study of these samples revealed the possibility to selectively remove protein nitration products during hemodialysis. Identification of intramolecular sites of nitration was preliminarily obtained in IgG chains isolated by 2D PAGE and assessed by bidimensional tandem mass spectrometry after chemoselective tagging. Further studies are needed to confirm at the molecular level the presence of nitrated Tyr residues in other proteins tentatively identified as nitration targets in this study and to explore the biological meaning of such a selective modification of plasma proteins by reactive nitrogen species in uremia and dialysis patients.

  • Identification of a new mutation in the gene coding for hairless protein responsible for alopecia universalis: The importance of direct gene sequencing.
    Publication Date: 15/01/2011, on Dermatology online journal
    by Nucara S, Colao E, Mangone G, Baudi F, Fabiani F, Nocera D, Passafaro G, Longo T, Laria AE, Malatesta P, Amato R, Trapasso F, Perrotti N
    DOI:

    Mutations in the gene HR coding for the hairless protein are associated with atrichia with papular lesions (APL), an autosomal recessive form of alopecia universalis that is characterized by generalized scalp and body atrichia with papular lesions. We here describe a South Italian family of ancient Albanian heritage. The full phenotype with complete atrichia was expressed in 2 siblings, whereas the parents and one sister were unaffected. Direct sequencing of the gene coding for the hairless protein allowed the identification of a new mutation in exon 17. Consistent with the recessive inheritance of the disease, both the siblings were homozygous for the mutation, whereas the parents and the unaffected sister where heterozygous. A relevant discrepancy with a haplotype linkage study is reported, stressing the importance of gene sequencing in genetic diagnosis and counseling because linkage studies can be biased by recombination events.

  • Proline derivatives in fruits of bergamot (Citrus bergamia Risso et Poit): presence of N-methyl-L-proline and 4-hydroxy-L-prolinebetaine.
    Publication Date: 12/01/2011, on Journal of agricultural and food chemistry
    by Servillo L, Giovane A, Balestrieri ML, Cautela D, Castaldo D
    DOI: 10.1021/jf102833v

    The content of proline and various compounds deriving from its metabolism (4-hydroxy-L-proline, N-methyl-L-proline, N,N-dimethylproline, and 4-hydroxy-L-prolinebetaine) was determined in fruits and seeds of Bergamot (Citrus bergamia Risso et Poit), growing in the Calabria region (South Italy). A HPLC-ESI-tandem mass spectrometry method, which allowed rapid determination of L-proline, 4-hydroxy-L-proline, N-methyl-L-proline, N,N-dimethylproline, and 4-hydroxy-L-prolinebetaine in juice and extracts of bergamot fruit with minimum sample preparation and short analysis time (about 10 min), is presented. Proline and 4-hydroxy-L-proline levels in the samples were also determined by HPLC analysis with fluorescence detection and the results compared to those obtained with HPLC-ESI-tandem mass spectrometry. For the first time, the presence of N-methyl-L-proline and 4-hydroxy-L-prolinebetaine in the fruits of a plant of the Citrus genus is reported.

  • Drawing with oblique coordinates: on a single case.
    Publication Date: 01/01/2011, on Behavioural neurology
    by Grossi D, Santangelo G, Carbone G, Giordano F, Angelillo VG, Trojano L
    DOI: 10.3233/BEN-2011-0333

    We describe a patient with right hemisphere damage affected by mild left visuo-spatial neglect and constructional apraxia. During the rehabilitation, he failed to draw a draught-board using horizontal and vertical trajectories, but he performed it successfully using oblique trajectories. These observations suggested an impairment of vertical/horizontal spatial coordinates system. In copying tasks including figure elements in different orientations he drew more accurately components in oblique orientation, whereas failed to reproduce components in horizontal orientation. The patient performed visuospatial perceptual and perceptual-imaginative tasks successfully. From these findings, it is possible to suggest that the oblique coordinate system of reference operates independently of vertical and horizontal coordinate systems in building a complex figure and that, therefore, cardinal orientation do not constitute a reference norm to define oblique orientation, as previously suggested.

  • Cognitive rehabilitation in non-communicative brain-damaged patients.
    Publication Date: 01/01/2011, on Functional neurology
    by Trojano L, Moretta P, Cozzolino A, Saltalamacchia A, Estraneo A
    DOI:

    Conscious patients with severe motor and speech disorders have great difficulty interacting with the environment and communicating with other people. Several augmentative communication devices are now available to exploit these patients' expressive potential, but their use often demands considerable cognitive effort. Non-communicative patients with severe brain lesions may have, in addition, specific cognitive deficits that hinder the efficient use of augmentative communication methods. Some neuropsychological batteries are now available for testing these patients. On the basis of such cognitive assessments, cognitive rehabilitation training can now be applied, but we underline that this training must be tailored to single patients in order to allow them to communicate autonomously and efficiently.

  • Knock-down of cathepsin D affects the retinal pigment epithelium, impairs swim-bladder ontogenesis and causes premature death in zebrafish.
    Publication Date: 01/01/2011, on PloS one
    by Follo C, Ozzano M, Mugoni V, Castino R, Santoro M, Isidoro C
    DOI: 10.1371/journal.pone.0021908

    The lysosomal aspartic protease Cathepsin D (CD) is ubiquitously expressed in eukaryotic organisms. CD activity is essential to accomplish the acid-dependent extensive or partial proteolysis of protein substrates within endosomal and lysosomal compartments therein delivered via endocytosis, phagocytosis or autophagocytosis. CD may also act at physiological pH on small-size substrates in the cytosol and in the extracellular milieu. Mouse and fruit fly CD knock-out models have highlighted the multi-pathophysiological roles of CD in tissue homeostasis and organ development. Here we report the first phenotypic description of the lack of CD expression during zebrafish (Danio rerio) development obtained by morpholino-mediated knock-down of CD mRNA. Since the un-fertilized eggs were shown to be supplied with maternal CD mRNA, only a morpholino targeting a sequence containing the starting ATG codon was effective. The main phenotypic alterations produced by CD knock-down in zebrafish were: 1. abnormal development of the eye and of retinal pigment epithelium; 2. absence of the swim-bladder; 3. skin hyper-pigmentation; 4. reduced growth and premature death. Rescue experiments confirmed the involvement of CD in the developmental processes leading to these phenotypic alterations. Our findings add to the list of CD functions in organ development and patho-physiology in vertebrates.

  • Structure-cytotoxicity relationships in bovine seminal ribonuclease: new insights from heat and chemical denaturation studies on variants.
    Publication Date: 01/01/2011, on The FEBS journal
    by Giancola C, Ercole C, Fotticchia I, Spadaccini R, Pizzo E, D'Alessio G, Picone D
    DOI: 10.1111/j.1742-4658.2010.07937.x

    Bovine seminal ribonuclease (BS-RNase), a homodimeric protein displaying selective cytotoxicity towards tumor cells, is isolated as a mixture of two isoforms, a dimeric form in which the chains swap their N-termini, and an unswapped dimer. In the cytosolic reducing environment, the dimeric form in which the chains swap their N-termini is converted into a noncovalent dimer (termed NCD), in which the monomers remain intertwined through their N-terminal ends. The quaternary structure renders the reduced protein resistant to the ribonuclease inhibitor, a protein that binds most ribonucleases with very high affinity. On the other hand, upon selective reduction, the unswapped dimer is converted in two monomers, which are readily bound and inactivated by the ribonuclease inhibitor. On the basis of these considerations, it has been proposed that the cytotoxic activity of BS-RNase relies on the 3D structure and stability of its NCD derivative. Here, we report a comparison of the thermodynamic and chemical stability of the NCD form of BS-RNase with that of the monomeric derivative, together with an investigation of the thermal dissociation mechanism revealing the presence of a dimeric intermediate. In addition, we report that the replacement of of Arg80 by Ser significantly decreases the cytotoxic activity of BS-RNase and the stability of the NCD form with respect to the parent protein, but does not affect the ribonucleolytic activity or the dissociation mechanism. The data show the importance of Arg80 for the cytotoxicity of BS-RNase, and also support the hypothesis that the reduced derivative of BS-RNase is responsible for its cytotoxic activity.

  • MicroRNA-restricted transgene expression in the retina.
    Publication Date: 01/01/2011, on PloS one
    by Karali M, Manfredi A, Puppo A, Marrocco E, Gargiulo A, Allocca M, Corte MD, Rossi S, Giunti M, Bacci ML, Simonelli F, Surace EM, Banfi S, Auricchio A
    DOI: 10.1371/journal.pone.0022166

    Gene transfer using adeno-associated viral (AAV) vectors has been successfully applied in the retina for the treatment of inherited retinal dystrophies. Recently, microRNAs have been exploited to fine-tune transgene expression improving therapeutic outcomes. Here we evaluated the ability of retinal-expressed microRNAs to restrict AAV-mediated transgene expression to specific retinal cell types that represent the main targets of common inherited blinding conditions.

  • Discrimination power of long-term heart rate variability measures for chronic heart failure detection.
    Publication Date: 01/01/2011, on Medical & biological engineering & computing
    by Melillo P, Fusco R, Sansone M, Bracale M, Pecchia L
    DOI: 10.1007/s11517-010-0728-5

    The aim of this study was to investigate the discrimination power of standard long-term heart rate variability (HRV) measures for the diagnosis of chronic heart failure (CHF). The authors performed a retrospective analysis on four public Holter databases, analyzing the data of 72 normal subjects and 44 patients suffering from CHF. To assess the discrimination power of HRV measures, an exhaustive search of all possible combinations of HRV measures was adopted and classifiers based on Classification and Regression Tree (CART) method was developed, which is a non-parametric statistical technique. It was found that the best combination of features is: Total spectral power of all NN intervals up to 0.4 Hz (TOTPWR), square root of the mean of the sum of the squares of differences between adjacent NN intervals (RMSSD) and standard deviation of the averages of NN intervals in all 5-min segments of a 24-h recording (SDANN). The classifiers based on this combination achieved a specificity rate and a sensitivity rate of 100.00 and 89.74%, respectively. The results are comparable with other similar studies, but the method used is particularly valuable because it provides an easy to understand description of classification procedures, in terms of intelligible "if … then …" rules. Finally, the rules obtained by CART are consistent with previous clinical studies.

  • Exercise dependence induced by pramipexole in Parkinson's Disease-a case report.
    Publication Date: 15/12/2010, on Movement disorders : official journal of the Movement Disorder Society
    by Vitale C, Santangelo G, Verde F, Amboni M, Sorrentino G, Grossi D, Barone P
    DOI: 10.1002/mds.23323

  • Design, synthesis and characterisation of a fluorescently labelled CyPLOS ionophore.
    Publication Date: 10/12/2010, on Chemistry (Weinheim an der Bergstrasse, Germany)
    by Coppola C, Paciello A, Mangiapia G, Licen S, Boccalon M, De Napoli L, Paduano L, Tecilla P, Montesarchio D
    DOI: 10.1002/chem.201000611

    A novel fluorescently labelled synthetic ionophore, based on a cyclic phosphate-linked disaccharide (CyPLOS) backbone and decorated with four tetraethylene glycol tails carrying dansyl units, has been synthesised in 12 steps in 26% overall yield. The key intermediate in the synthetic strategy is a novel glucoside building block, serving through its 2- and 3-hydroxy groups as the anchor point for flexible tetraethylene glycol tentacles with reactive azido moieties at their ends. To test the versatility of this glucoside scaffold, it was preliminarily functionalised with a set of diverse probes--as fluorescent, redox-active or hydrophobic tags--either by reduction of the azides followed by condensation with activated carboxylic acid derivatives, or by a direct coupling with a terminal alkyne in a Cu(I)-promoted 1,3-dipolar cycloaddition. Tagging of the monomeric building block with dansyl residues allowed us to prepare a fluorescent, amphiphilic macrocycle, which was investigated for its propensity to self-aggregate in CDCl(3)--studied by means of concentration-dependent (31)P NMR spectroscopy experiments--and in aqueous solution, in which combined dynamic light scattering (DLS) and small-angle neutron scattering (SANS) measurements provided a detailed physico-chemical analysis of the self-assembled systems, mainly organised in the form of large vesicles. Its ion-transport properties through phospholipid bilayers, determined by HPTS fluorescence assays, showed this compound to be more active than the previously synthesised CyPLOS congeners. Solvent-dependent fluorescence changes for the labelled ionophore in liposome suspension established that the dansyl moieties are dispersed in environments with polarity intermediate between those of CH(2)Cl(2) and propan-2-ol, suggesting that the CyPLOS tentacles infiltrate the mid-polar region of the membranes.

  • Women survive breast cancer but fall victim to heart failure: the shadows and lights of targeted therapy.
    Publication Date: 01/12/2010, on Journal of cardiovascular medicine (Hagerstown, Md.)
    by Maurea N, Coppola C, Ragone G, Frasci G, Bonelli A, Romano C, Iaffaioli RV
    DOI: 10.2459/JCM.0b013e328336b4c1

    In many cases, early-stage breast cancer is now curable, and metastatic disease can be chronic consequent to the advent of new therapeutic tools. Unfortunately, some treatments have been associated with adverse cardiovascular effects. Indeed, in many breast cancer survivors, the risk of cardiovascular disease is higher than the risk of cancer recurrence. The clinical challenge of preventing cardiovascular complications in patients undergoing antineoplastic treatment has two aims, more effective life-saving treatment of patients, and prevention of morbidity and cardiovascular mortality in the short term and long term. The aim of the present study is to review the rapidly evolving therapeutic strategies designed to treat early-stage breast cancer. The review highlights the need for more data on the impact of new biological drugs (targeted therapy) on the cardiovascular apparatus. Finally, given the complexity of targeted and other novel treatments, cancer patients are best managed through a multidisciplinary approach.

  • Physico-chemical features of the environment affect the protein conformation and the immunoglobulin E reactivity of kiwellin (Act d 5).
    Publication Date: 01/12/2010, on Clinical and experimental allergy : journal of the British Society for Allergy and Clinical Immunology
    by Bernardi ML, Picone D, Tuppo L, Giangrieco I, Petrella G, Palazzo P, Ferrara R, Tamburrini M, Mari A, Ciardiello MA
    DOI: 10.1111/j.1365-2222.2010.03603.x

    Allergy diagnostic systems sometimes give false positive or negative results. In this respect, the influence of protein conformational changes on the allergen-IgE interaction sites is worthy to be investigated.

  • DWI Reversibility after Intra-Arterial Thrombolysis. A Case Report and Literature Review.
    Publication Date: 01/12/2010, on The neuroradiology journal
    by Tortora F, Cirillo M, Ferrara M, Manto A, Briganti F, Cirillo S
    DOI: 10.1177/197140091002300618

    We report our case and review the literature on reversal DWI lesions, ADC thresholds and correlation between DWI lesion and outcome measured with clinical scales. A 30-years old woman was admitted to our hospital 18 hours after stroke onset. Considering the absence of alterations on CT and the worsening of symptomatology, the patient underwent MRI, which showed a slightly hyperintense signal in FLAIR images in the left portion of the pons and midbrain and a more evident bilateral DWI hyperintensity of the pons. The patient was treated with mechanical and pharmacological intra-arterial thrombolysis. The patient showed a rapid improvement of symptoms. Two weeks after the treatment her clinical conditions were characterized by a residual right hemiparesis and complete recovery of right motility, respiratory and swallowing difficulties. MR examination demonstrated a slight signal alteration of the pons left hemiportion and a disappearance of the mesencephalic signal alteration and of the right portion of the pons. DWI lesions represent irreversibly damaged tissue but new evidence suggests that DWI lesions may be reversible, especially with reperfusion, by now well demonstrated in animal models. Therefore acute DWI lesions probably contain not only irreversibly injured tissue but also parts of the penumbra. The debate on the capability of ADC maps to discriminate irreversibly from reversibly damaged tissue is a matter of controversy. ADC values in human stoke are not an independent indicator of tissue viability. The use of thresholds may improve reproducibility but not validity.

  • Role of Magnetic Resonance Imaging in Duane's Retraction Syndrome: Presence of the Abducens Nerve Depending on Type. A Clinical-Anatomical Study.
    Publication Date: 01/12/2010, on The neuroradiology journal
    by Taglialatela G, Conforti R, Notaro M, Cotticelli L, Caranci F, Cirillo S
    DOI: 10.1177/197140091002300610

    This study correlated anatomical findings and clinical evidence in Duane's Retraction Syndrome using brain MRI. The study included 11 patients with different types of Duane's Retraction Syndrome (DRS). Each patient underwent brain MRI. The VI(th) nerve was absent in all patients with DRS-I, present in all patients with DRS-II, while findings varied in patients with DRS-III. Our study confirms the literature on the usefulness of brain MRI in diagnosis of Duane's Retraction Syndrome in children.