Impact of phytosterols on liver and distal colon metabolome in experimental murine colitis model: an explorative study.
Publication Date: 01/12/2019, on Journal of enzyme inhibition and medicinal chemistry
by Iaccarino N, Amato J, Pagano B, Di Porzio A, Micucci M, Bolelli L, Aldini R, Novellino E, Budriesi R, Randazzo A
Phytosterols are known to reduce plasma cholesterol levels and thereby reduce cardiovascular risk. Studies conducted on human and animal models have demonstrated that these compounds have also anti-inflammatory effects. Recently, an experimental colitis model (dextran sulphate sodium-induced) has shown that pre-treatment with phytosterols decreases infiltration of inflammatory cells and accelerates mucosal healing. This study aims to understand the mechanism underlying the colitis by analysing the end-products of the metabolism in distal colon and liver excised from the same mice used in the previous work. In particular, an unsupervised gas chromatography-mass spectrometry (GC-MS) and NMR based metabolomics approach was employed to identify the metabolic pathways perturbed by the dextran sodium sulphate (DSS) insult (i.e. Krebs cycle, carbohydrate, amino acids, and nucleotide metabolism). Interestingly, phytosterols were able to restore the homeostatic equilibrium of the hepatic and colonic metabolome.
Clustering in the Golgi apparatus governs sorting and function of GPI-APs in polarized epithelial cells.
Publication Date: 10/08/2019, on FEBS letters
by Lebreton S, Paladino S, Zurzolo C
Glycosylphosphatidylinositol-anchored proteins (GPI-APs) are lipid-anchored proteins attached to the extracellular leaflet of the plasma membrane via a glycolipid anchor. GPI-APs are commonly associated with cholesterol- and sphingolipid-enriched membrane microdomains. These microdomains help regulating various biological activities, by segregating different proteins and lipids in (nanoscale) membrane compartments. In fibroblasts, GPI-APs form actin- and cholesterol-dependent nanoclusters directly at the plasma membrane (PM). By contrast, in polarized epithelial cells GPI-APs cluster in the Golgi apparatus, the major protein-sorting hub for the secretory pathway. Golgi clustering is required for the selective sorting of GPI-APs to the apical PM domain, but also regulates their organization and biological activities at the cell surface. In this review, we discuss recent advances in our understanding of the mechanism of GPI-AP sorting to the apical membrane. We focus on the roles of the protein moiety, lipids, and calcium ions in the regulation of the clustering of GPI-APs in the Golgi apparatus. This article is protected by copyright. All rights reserved.
A comparative assessment of metals and phthalates in commercial tea infusions: A starting point to evaluate their tolerance limits.
Publication Date: 01/08/2019, on Food chemistry
by Troisi J, Richards S, Symes S, Ferretti V, Di Maio A, Amoresano A, Daniele B, Aliberti F, Guida M, Trifuoggi M, De Castro O
Tea is one of the most consumed beverages in the word. Here we report the concentrations of metals and phthalates in 32 commercial tea packages. The data were used to estimate the average daily intake of metals and phthalates, and associated Hazard Quotients (HQ) were calculated in order to determine risk of non-cancerous health effects for adults consuming tea on a daily basis. Tea samples were chosen based on the sales network, the price, the marketing quality and the presence of filters in the packages. Relatively high median concentrations of Al (5240 µg/L), Ni (44 µg/L), and Mn (2919 µg/L) were detected. No metals or phthalates quantified in the tea infusions and soluble tea showed an HQ greater than 1, indicating no risk of non-cancerous health effects. The data presented herein may serve as a starting point to evaluate tolerance limits of metals and phthalate in the tea infusion.
A physicochemical investigation on the metal binding properties of TtSmtB, a thermophilic member of the ArsR/SmtB transcription factor family.
Publication Date: 26/07/2019, on International journal of biological macromolecules
by Gallo G, Antonucci I, Pirone L, Amoresano A, Contursi P, Limauro D, Pedone E, Bartolucci S, Fiorentino G
The transcription factors of the ArsR/SmtB family are widespread within the bacterial and archaeal kingdoms. They are transcriptional repressors able to sense a variety of metals and undergo allosteric conformational changes upon metal binding, resulting in derepression of genes involved in detoxification. So far, the molecular determinants of specificity, selectivity, and metal binding mechanism have been scarcely investigated in thermophilic microorganisms. TtSmtB, the only ArsR/SmtB member present in the genome of Thermus thermophilus HB27, was chosen as a model to shed light into such molecular mechanisms at high temperature. In the present study, using a multidisciplinary approach, a structural and functional characterization of the protein was performed focusing on its metal interaction and chemical-physical stability. Our data demonstrate that TtSmtB has two distinct metal binding sites per monomer and interacts with di-tri-penta-valent ions with different affinity. Detailed knowledge at molecular level of protein-metal interaction is remarkable to design metal binding domains as scaffolds in metal-based therapies as well as in metal biorecovery or biosensing in the environment.
Enactment effect in patients with Alzheimer's disease.
Publication Date: 16/07/2019, on Journal of clinical and experimental neuropsychology
by De Lucia N, Milan G, Conson M, Grossi D, Trojano L
: Subjects can improve their performance on memory for action phrases if, during the encoding condition, they self-perform actions associated with verbs (subject-performed condition), or if they perceive the actions carried out by experimenter (experimenter-performed condition), with respect to a verbal task condition in which they only read or listen to the stimuli. This facilitation is labeled "Enactment effect" (EE), and is thought to be associated with episodic integration processes binding actions and nouns together in a coherent representation. Only recently, studies addressed EE in AD individuals reporting significant improvements on memory tasks in the subject-performed encoding condition. However, no studies tried to explore the cognitive mechanisms supporting EE in AD individuals. : Performance on recognition and cued recall tasks for action phrases were assessed in a sample of 32 mild-to-moderate AD individuals and 30 healthy adults, in verbal, subject-performed and experimenter-performed encoding conditions. Moreover, a cognitive assessment was completed to explore the possible correlates of EE in our participants. : Results showed that both subject-performed and experimenter-performed encoding conditions produced similar advantages over the verbal condition, in both memory tasks in both groups. Moreover, these memory advantages were strongly associated to executive processes, in both AD and healthy adults. : The present study confirmed that EE is spared in mild to moderate AD. Our findings supported the role of episodic integration processes and suggested a contribution of executive processes in EE.
High openness and high extroversion are linked with better time-based prospective memory in multiple sclerosis.
Publication Date: 16/07/2019, on Journal of neurology
by Raimo S, Trojano L, Gaita M, Spitaleri D, Santangelo G
Prospective memory (PM) deficits are often reported in multiple sclerosis (MS), but their relationship with neuropsychological characteristics and personality traits remains to be explored.
MicroRNA-33 and SIRT1 influence the coronary thrombus burden in hyperglycemic STEMI patients.
Publication Date: 11/07/2019, on Journal of cellular physiology
by D'Onofrio N, Sardu C, Paolisso P, Minicucci F, Gragnano F, Ferraraccio F, Panarese I, Scisciola L, Mauro C, Rizzo MR, Mansueto G, Varavallo F, Brunitto G, Caserta R, Tirino V, Papaccio G, Barbieri M, Paolisso G, Balestrieri ML, Marfella R
Primary percutaneous coronary intervention (PPCI) is a pivotal treatment in ST-segment elevation myocardial infarction (STEMI) patients. However, in hyperglycemic-STEMI patients, the incidence of death is still significant. Here, the involvement of sirtuin 1 (SIRT1) and miR33 on the pro-inflammatory/pro-coagulable state of the coronary thrombus was investigated. Moreover, 1-year outcomes in hyperglycemic STEMI in patients subjected to thrombus aspiration before PPCI were evaluated. Results showed that hyperglycemic thrombi displayed higher size and increased miR33, reactive oxygen species, and pro-inflammatory/pro-coagulable markers. Conversely, the hyperglycemic thrombi showed a lower endothelial SIRT1 expression. Moreover, in vitro experiments on endothelial cells showed a causal effect of SIRT1 modulation on the pro-inflammatory/pro-coagulative state via hyperglycemia-induced miR33 expression. Finally, SIRT1 expression negatively correlated with STEMI outcomes. These observations demonstrate the involvement of the miR33/SIRT1 pathway in the increased pro-inflammatory and pro-coagulable state of coronary thrombi in hyperglycemic STEMI patients.
The microRNA-29a Modulates Serotonin 5-HT7 Receptor Expression and Its Effects on Hippocampal Neuronal Morphology.
Publication Date: 10/07/2019, on Molecular neurobiology
by Volpicelli F, Speranza L, Pulcrano S, De Gregorio R, Crispino M, De Sanctis C, Leopoldo M, Lacivita E, di Porzio U, Bellenchi GC, Perrone-Capano C
miRNAs are master regulators of gene expression in diverse biological processes, including the modulation of neuronal cytoarchitecture. The identification of their physiological target genes remains one of the outstanding challenges. Recently, it has been demonstrated that the activation of serotonin receptor 7 (5-HT7R) plays a key role in regulating the neuronal structure, synaptogenesis, and synaptic plasticity during embryonic and early postnatal development of the central nervous system (CNS). In order to identify putative miRNAs targeting the 3'UTR of 5-HT7R mouse transcript, we used a computational prediction tool and detected the miR-29 family members as the only candidates. Thus, since miR-29a is more expressed than other members in the brain, we investigated its possible involvement in the regulation of neuronal morphology mediated by 5-HT7R. By luciferase assay, we show that miR-29a can act as a post-transcriptional regulator of 5-HT7R mRNA. Indeed, it downregulates 5-HT7R gene expression in cultured hippocampal neurons, while the expression of other serotonin receptors is not affected. From a functional point of view, miR-29a overexpression in hippocampal primary cultures impairs the 5HT7R-dependent neurite elongation and remodeling through the inhibition of the ERK intracellular signaling pathway. In vivo, the upregulation of miR-29a in the developing hippocampus parallels with the downregulation of 5-HT7R expression, supporting the hypothesis that this miRNA is a physiological modulator of 5-HT7R expression in the CNS.
Bio-Inspired Dual-Selective <i>BCL-2</i>/<i>c-MYC</i> G-Quadruplex Binders: Design, Synthesis, and Anticancer Activity of Drug-like Imidazo[2,1-<i>i</i>]purine Derivatives.
Publication Date: 10/07/2019, on Journal of medicinal chemistry
by Pelliccia S, Amato J, Capasso D, Di Gaetano S, Massarotti A, Piccolo M, Irace C, Tron GC, Pagano B, Randazzo A, Novellino E, Giustiniano M
In the search for new drug-like selective G-quadruplex binders, a bioinspired design focused on the use of nucleobases as synthons in a multicomponent reaction was herein proved to be viable and successful. Hence, a new class of multifunctionalized imidazo[2,1-]purine derivatives, easily synthesized with a convergent approach, allowed for the identification of the first dual / gene promoter G-quadruplex ligand. Biophysical studies involving circular dichroism melting experiments, microscale thermophoresis measurements, NMR titrations, and computational docking calculations, as well as biological investigations including cytotoxicity and apoptotic assays, and quantitative polymerase chain reaction and Western blot analyses, were performed to assess the potency and to characterize the binding mode of the newly identified lead compound. The absence of toxicity toward normal cells, together with the small molecular weight (≅500 Da), the water solubility, the ease of functionalization, and the selectivity profile, are promising and desirable features to develop G-quadruplex binders as safe and effective anticancer agents.
Insights into the G-rich VEGF-binding aptamer V7t1: when two G-quadruplexes are better than one!
Publication Date: 05/07/2019, on Nucleic acids research
by Moccia F, Riccardi C, Musumeci D, Leone S, Oliva R, Petraccone L, Montesarchio D
The G-quadruplex-forming VEGF-binding aptamer V7t1 was previously found to be highly polymorphic in a K+-containing solution and, to restrict its conformational preferences to a unique, well-defined form, modified nucleotides (LNA and/or UNA) were inserted in its sequence. We here report an in-depth biophysical characterization of V7t1 in a Na+-rich medium, mimicking the extracellular environment in which VEGF targeting should occur, carried out combining several techniques to analyse the conformational behaviour of the aptamer and its binding to the protein. Our results demonstrate that, in the presence of high Na+ concentrations, V7t1 behaves in a very different way if subjected or not to annealing procedures, as evidenced by native gel electrophoresis, size exclusion chromatography and dynamic light scattering analysis. Indeed, not-annealed V7t1 forms both monomeric and dimeric G-quadruplexes, while the annealed oligonucleotide is a monomeric species. Remarkably, only the dimeric aptamer efficiently binds VEGF, showing higher affinity for the protein compared to the monomeric species. These findings provide new precious information for the development of improved V7t1 analogues, allowing more efficient binding to the cancer-related protein and the design of effective biosensors or theranostic devices based on VEGF targeting.
Association of human leukocyte antigen-G 14 bp polymorphism with recurrent pregnancy loss in European countries: a meta-analysis of literature studies.
Publication Date: 04/07/2019, on Fertility and sterility
by Monti M, Lupoli R, Sosa Fernandez LM, Cirillo F, Di Minno MND
To study the controversial association between human leukocyte antigen-G (HLA-G) 14 bp polymorphism and recurrent pregnancy loss (RPL). We performed a meta-analysis of studies in the literature that enrolled only women of European countries who experienced RPL spontaneously or after undergoing IVF.
Characterization of a Surface-Active Protein Extracted from a Marine Strain of <i>Penicillium chrysogenum</i>.
Publication Date: 02/07/2019, on International journal of molecular sciences
by Cicatiello P, Stanzione I, Dardano P, De Stefano L, Birolo L, De Chiaro A, Monti DM, Petruk G, D'Errico G, Giardina P
Marine microorganisms represent a reservoir of new promising secondary metabolites. Surface-active proteins with good emulsification activity can be isolated from fungal species that inhabit the marine environment and can be promising candidates for different biotechnological applications. In this study a novel surface-active protein, named Sap-, was purified from a marine strain of The effect of salt concentration and temperature on protein production was analyzed, and a purification method was set up. The purified protein, identified as Pc13g06930, was annotated as a hypothetical protein. It was able to form emulsions, which were stable for at least one month, with an emulsification index comparable to that of other known surface-active proteins. The surface tension reduction was analyzed as function of protein concentration and a critical micellar concentration of 2 μM was determined. At neutral or alkaline pH, secondary structure changes were monitored over time, concurrently with the appearance of protein precipitation. Formation of amyloid-like fibrils of SAP was demonstrated by spectroscopic and microscopic analyses. Moreover, the effect of protein concentration, a parameter affecting kinetics of fibril formation, was investigated and an on-pathway involvement of micellar aggregates during the fibril formation process was suggested.
Full-field electroretinography, visual acuity and visual fields in Usher syndrome: a multicentre European study.
Publication Date: 02/07/2019, on Documenta ophthalmologica. Advances in ophthalmology
by Stingl K, Kurtenbach A, Hahn G, Kernstock C, Hipp S, Zobor D, Kohl S, Bonnet C, Mohand-Saïd S, Audo I, Fakin A, Hawlina M, Testa F, Simonelli F, Petit C, Sahel JA, Zrenner E
Usher syndrome (USH) is a multisensory deficiency involving vision, hearing and the vestibular system. The purpose of this study is to report on the functional data (i.e. electroretinography, visual fields, visual acuity) of patients with retinitis pigmentosa (RP) due to Usher syndrome that were collected in a multicentre European study (TREATRUSH).
Prospective memory in Parkinson's disease: the role of the motor subtypes.
Publication Date: 29/06/2019, on Journal of neurology
by D'Iorio A, Maggi G, Vitale C, Amboni M, Di Meglio D, Trojano L, Santangelo G
Prospective memory (PM) is defined as memory for future intentions and it is typically divided into time-based and event-based PM. Deficit of PM has been reported in patients with Parkinson's disease (PD) but no study has yet explored the association between motor subtypes (tremor dominant and rigidity/bradykinesia dominant) and performance on PM tasks. The aim of the study was to explore the role of motor subtypes in the defect of PM.
Pioglitazone Improves Mitochondrial Organization and Bioenergetics in Down Syndrome Cells.
Publication Date: 28/06/2019, on Frontiers in genetics
by Mollo N, Nitti M, Zerillo L, Faicchia D, Micillo T, Accarino R, Secondo A, Petrozziello T, Calì G, Cicatiello R, Bonfiglio F, Sarnataro V, Genesio R, Izzo A, Pinton P, Matarese G, Paladino S, Conti A, Nitsch L
Mitochondrial dysfunction plays a primary role in neurodevelopmental anomalies and neurodegeneration of Down syndrome (DS) subjects. For this reason, targeting mitochondrial key genes, such as , is emerging as a good therapeutic approach to attenuate cognitive disability in DS. After demonstrating the efficacy of the biguanide metformin (a activator) in a cell model of DS, we extended the study to other molecules that regulate the pathway acting on genes. We, therefore, treated trisomic fetal fibroblasts with different doses of pioglitazone (PGZ) and evaluated the effects on mitochondrial dynamics and function. Treatment with PGZ significantly increased mRNA and protein levels of PGC-1α. Mitochondrial network was fully restored by PGZ administration affecting the fission-fusion mitochondrial machinery. Specifically, optic atrophy 1 () and mitofusin 1 () were upregulated while dynamin-related protein 1 () was downregulated. These effects, together with a significant increase of basal ATP content and oxygen consumption rate, and a significant decrease of reactive oxygen species (ROS) production, provide strong evidence of an overall improvement of mitochondria bioenergetics in trisomic cells. In conclusion, we demonstrate that PGZ is able to improve mitochondrial phenotype even at low concentrations (0.5 μM). We also speculate that a combination of drugs that target mitochondrial function might be advantageous, offering potentially higher efficacy and lower individual drug dosage.