Role of the H-bond between L53 and T56 for Aquaporin-4 epitope in Neuromyelitis Optica

Aquaporin-4 (AQP4) is the CNS water channel organized into well-ordered protein aggregates called Orthogonal Arrays of Particles (OAPs). Neuromyelitis Optica (NMO) is an autoimmune disease caused by anti-OAP autoantibodies (AQP4-IgG). Molecular Dynamics (MD) simulations have identified an H-bond between L53 and T56 as the key for AQP4 epitope and therefore of potential interest for drug design in NMO field. In the present study, we have experimentally tested this MD-prediction using the classic mutagenesis approach. We substituted T56 with V56 and tested this mutant for AQP4 aggregates and AQP4-IgG binding. gSTED super-resolution microscopy showed that the mutation does not affect AQP4 aggregate dimension; immunofluorescence and cytofluorimetric analysis demonstrated its unaltered AQP4-IgG binding, therefore invalidating the MD-prediction. We later investigated whether AQP4, expressed in Sf9 insect and HEK-293F cells, is able to correctly aggregate before and after the purification steps usually applied to obtain AQP4 crystal. The results demonstrated that AQP4-IgG recognizes AQP4 expressed in Sf9 and HEK-293F cells by immunofluorescence even though BN-PAGE analysis showed that AQP4 forms smaller aggregates when expressed in insect cells compared to mammalian cell lines. Notably, after AQP4 purification, from both insect and HEK-293F cells, no aggregates are detectable by BN-PAGE and AQP4-IgG binding is impaired in sandwich ELISA assays. All together these results indicate that 1) the MD prediction under analysis is not supported by experimental data and 2) the procedure to obtain AQP4 crystals might affect its native architecture and, as a consequence, MD simulations. In conclusion, given the complex nature of the AQP4 epitope, MD might not be the suitable for molecular medicine advances in NMO

AQP4-independent TRPV4 modulation of plasma membrane water permeability

: Despite of the major role of aquaporin (AQP) water channels in controlling transmembrane water fluxes, alternative ways for modulating water permeation have been proposed. In the Central Nervous System (CNS), Aquaporin-4 (AQP4) is reported to be functionally coupled with the calcium-channel Transient-Receptor Potential Vanilloid member-4 (TRPV4), which is controversially involved in cell volume regulation mechanisms and water transport dynamics. The present work aims to investigate the selective role of TRPV4 in regulating plasma membrane water permeability in an AQP4-independent way. Fluorescence-quenching water transport experiments in Aqp4-/- astrocytes revealed that cell swelling rate is significantly increased upon TRPV4 activation and in the absence of AQP4. The biophysical properties of TRPV4-dependent water transport were therefore assessed using the HEK-293 cell model. Calcein quenching experiments showed that chemical and thermal activation of TRPV4 overexpressed in HEK-293 cells leads to faster swelling kinetics. Stopped-flow light scattering water transport assay was used to measure the osmotic permeability coefficient (Pf, cm/s) and activation energy (Ea, kcal/mol) conferred by TRPV4. Results provided evidence that although the Pf measured upon TRPV4 activation is lower than the one obtained in AQP4-overexpressing cells (Pf of AQP4 = 0.01667 ± 0.0007; Pf of TRPV4 = 0.002261 ± 0.0004; Pf of TRPV4 + 4αPDD = 0.007985 ± 0.0006; Pf of WT = 0.002249 ± 0.0002), along with activation energy values (Ea of AQP4 = 0.86 ± 0.0006; Ea of TRPV4 + 4αPDD = 2.73 ± 1.9; Ea of WT = 8.532 ± 0.4), these parameters were compatible with a facilitated pathway for water movement rather than simple diffusion. The possibility to tune plasma membrane water permeability more finely through TRPV4 might represent a protective mechanism in cells constantly facing severe osmotic challenges to avoid the potential deleterious effects of the rapid cell swelling occurring via AQP channels

Group B Streptococcus early-onset disease and observation of well-appearing newborns

Background International guidelines lack a substantial consensus regarding management of asymptomatic full-term and late preterm neonates at risk for early-onset disease (EOS). Large cohorts of newborns are suitable to increase the understanding of the safety and efficacy of a given strategy. Methods This is a prospective, area-based, cohort study involving regional birth facilities of Emilia-Romagna (Italy). We compared cases of EOS (at or above 35 weeks\u2019 gestation) registered in 2003\u20132009 (baseline period: 266,646 LBs) and in 2010\u20132016, after introduction of a new strategy (serial physical examinations, SPEs) for managing asymptomatic neonates at risk for EOS (intervention period: 265,508 LBs). Results There were 108 cases of EOS (baseline period, n = 60; intervention period, n = 48). Twenty-two (20.4%) remained asymptomatic through the first 72 hours of life, whereas 86 (79.6%) developed symptoms, in most cases (52/86, 60.5%) at birth or within 6 hours. The median age at presentation was significantly earlier in the intrapartum antibiotic prophylaxis (IAP)exposed than in the IAP-unexposed neonates (0 hours, IQR 0.0000\u20130.0000 vs 6 hours, IQR 0.0000\u201315.0000, p<0.001). High number of neonates (n = 531) asymptomatic at birth, exposed to intrapartum fever, should be treated empirically for each newborn who subsequently develops sepsis. IAP exposed neonates increased (12% vs 33%, p = 0.01), age at presentation decreased (median 6 vs 1 hours, p = 0.01), whereas meningitis, mechanical ventilation and mortality did not change in baseline vs intervention period. After implementing the SPEs, no cases had adverse outcomes due to the strategy, and no cases developed severe disease after 6 hours of life. Conclusions Infants with EOS exposed to IAP developed symptoms at birth in almost all cases, and those who appeared well at birth had a very low chance of having EOS. The risk of EOS in neonates (asymptomatic at birth) exposed to intrapartum fever was low. Although definite conclusions on causation are lacking, our data support SPEs of asymptomatic newborns at risk for EOS. SPEs seems a safe and effective alternative to laboratory screening and empirical antibiotic therapy

Annali storici di Principato Citra, A. 6, n. 2.2 (2008)

A. 6, n. 2.2 (2008): G. Guardia, Editoriale, P. 3 ; A. Trezza, Il Cilento in semiotica del paesaggio: testo della cultura e testo paesaggistico, P. 5 ; M.L. Mautone, Un cofanetto nuziale della bottega degli Embrachi al Museo Diocesano di Vallo della Lucania, P. 35 ; E. Granito, Lotte operaie nel Salernitano dall’Unità all’Età Giolittiana. Menechella la scapricciata e i suoi compagni, P. 48 ; A. Gargano, Affreschi oligarchici: lettere anonime, ricorsi e inchieste prefettizie nella vita socio-amministrativa di Pagani nel tardo Ottocento, P. 69 ; E. Bianco, L’incastellamento medievale nel Cilento interno: il borgo murato di Stio (XI-XVI sec.). Prime indagini, P. 96 ; E. Frescani, L’arte di imparare. Note sull’apprendistato nella valle dell’Irno nel XVII secolo, P. 109 ; E. Catone, I marchesi Forcella a Buccino. Brevi note genealogiche, P. 114 ; G. Pepe, Francesco Durelli: uno storico ignorato, P. 124 ; A. La Greca, “In diem Christi conscientiam onerantes”. L’atto di rifondazione della confraternita del SS. Rosario di Montecorice e del suo trasferimento dalla cappella di S. Sofia alla nuova chiesa parrocchiale di S. Biagio (6 febbraio 1589), P. 139 ; A. Capano, Note di toponomastica in margine al Catasto provvisorio di San Giovanni a Piro (1815) e di Bosco sua frazione (20 sett. 1828), P. 146 ; Nota di Redazione, Pioppi: « culla » della Dieta Mediterranea. Celebrati i 25 anni di “Cronache Cilentane” e la XVIII Edizione degli “Incontri Mediterranei”, P. 180 ; M. Serra, Un’avvincente iniziativa editoriale del Centro di Promozione Culturale per il Cilento, P. 183 ; G. De Vita, “Oltre la torre d’avorio”. Un libro degli allievi per i sessant’anni del prof. Luigi Rossi, P. 188

Serum Albumin Is Inversely Associated With Portal Vein Thrombosis in Cirrhosis

We analyzed whether serum albumin is independently associated with portal vein thrombosis (PVT) in liver cirrhosis (LC) and if a biologic plausibility exists. This study was divided into three parts. In part 1 (retrospective analysis), 753 consecutive patients with LC with ultrasound-detected PVT were retrospectively analyzed. In part 2, 112 patients with LC and 56 matched controls were entered in the cross-sectional study. In part 3, 5 patients with cirrhosis were entered in the in vivo study and 4 healthy subjects (HSs) were entered in the in vitro study to explore if albumin may affect platelet activation by modulating oxidative stress. In the 753 patients with LC, the prevalence of PVT was 16.7%; logistic analysis showed that only age (odds ratio [OR], 1.024; P = 0.012) and serum albumin (OR, -0.422; P = 0.0001) significantly predicted patients with PVT. Analyzing the 112 patients with LC and controls, soluble clusters of differentiation (CD)40-ligand (P = 0.0238), soluble Nox2-derived peptide (sNox2-dp; P < 0.0001), and urinary excretion of isoprostanes (P = 0.0078) were higher in patients with LC. In LC, albumin was correlated with sCD4OL (Spearman's rank correlation coefficient [r(s)], -0.33; P < 0.001), sNox2-dp (r(s), -0.57; P < 0.0001), and urinary excretion of isoprostanes (r(s), -0.48; P < 0.0001) levels. The in vivo study showed a progressive decrease in platelet aggregation, sNox2-dp, and urinary 8-iso prostaglandin F2 alpha-III formation 2 hours and 3 days after albumin infusion. Finally, platelet aggregation, sNox2-dp, and isoprostane formation significantly decreased in platelets from HSs incubated with scalar concentrations of albumin. Conclusion: Low serum albumin in LC is associated with PVT, suggesting that albumin could be a modulator of the hemostatic system through interference with mechanisms regulating platelet activation

Visita all'Osservatorio della Biodiversità marina e terrestre della Regione Sicilia - ORBS

Con il taglio inaugurale del nastro il 16 dicembre 2015, prende vita la struttura museale permanente dell'Osservatorio della Biodiversità marina e terrestre della Regione Sicilia che porta lo stesso nome del Progetto di Ricerca "ORBS – Sistema di comunicazione, informazione e diffusione dell'Osservatorio Regionale della Sicilia", intitolata il 21 dicembre 2018 al Dott. Sandro Fiorelli. Ad oggi, la struttura, è operativa presso la Sede Secondaria dell'Istituto per lo studio degli impatti Antropici e Sostenibilità in ambiente marino del Consiglio Nazionale delle Ricerche (IAS – CNR) di Capo Granitola. Il progetto ORBS, finanziato da Regione Siciliana - Assessorato alla Cooperazione, Commercio, Artigianato e Pesca - Dipartimento Pesca, con periodo di attività 2013 - 2015, si è concluso proprio con la realizzazione della struttura museale; l'Osservatorio è stato istituito dall'Assessorato del Territorio e dell'Ambiente della Regione Siciliana nell'ambito di un accordo quadro con ARPA, ISPRA e CNR. Grazie al progetto ORBS, docenti e allievi dell'Accademia di Belle Arti di Palermo e il personale CNR – IAS (ex IAMC) S. S. di Capo Granitola, hanno collaborato sinergicamente permettendo di realizzare delle azioni didattiche e creative di valore scientifico espresse con straordinaria forza e bellezza. Ricercatori e professori si sono confrontati al fine di combinare le proprie competenze riuscendo nel progetto ambizioso di coinvolgere e fondere i diversi ambiti scientifici sensibilizzando gli artisti ai temi della Biodiversità. Le opere prodotte, corredate da schede scientifiche, hanno oltre al valore artistico un aggiunto valore didattico. L'apertura della sezione espositiva dedicata alla diffusione e alla comunicazione della biodiversità rappresenta da un lato l'importante tappa conclusiva del progetto, dall'altro l'inizio di un percorso mirato alla diffusione della biodiversità verso il mondo giovanile, le scuole e per tutto il territorio. Questa strepitosa collaborazione "CNR – Accademia di Belle Arti di Palermo" conferma l'importanza e l'opportunità di unire arte e scienza per esaltare la percezione della ricerca scientifica da parte della comunità. La divulgazione della scienza è un'attività complessa e sicuramente necessita di competenze e attitudini multidisciplinari oltreché di motivazione ed entusiasmo. La comunicazione delle tematiche scientifiche, di per sé ardua nella traduzione al grande pubblico, grazie alla forza esplicativa dell'arte, diviene opportunità di riflessione, osservazione, confronto per le comunità di visitatori. Il coordinamento delle visite delle scuole di ogni ordine e grado, Enti Pubblici, Comunità Scientifica, Cariche Istituzionali, Delegazioni di Politici Italiani e Stranieri, Associazioni Culturali, Associazioni No-Profit di Volontariato, Associazioni di Promozione Sociale, Organizzazioni di Volontariato, Onlus, pubblico in generale, presso ORBS, è affidato al qualificato personale (tecnici, tecnologi e ricercatori) dell'IAS – CNR S. S. di Capo Granitola, che gestisce in prima persona i visitatori nel percorso didattico e promuove il valore della divulgazione scientifica perseguendo la terza missione degli Enti di Ricerca, attraverso l'applicazione diretta, la valorizzazione e l'impiego della conoscenza

Multi-wavelength observations of blazar AO 0235+164 in the 2008-2009 flaring state

The blazar AO 0235+164 (z = 0.94) has been one of the most active objects observed by Fermi Large Area Telescope (LAT) since its launch in Summer 2008. In addition to the continuous coverage by Fermi, contemporaneous observations were carried out from the radio to γ-ray bands between 2008 September and 2009 February. In this paper, we summarize the rich multi-wavelength data collected during the campaign (including F-GAMMA, GASP-WEBT, Kanata, OVRO, RXTE, SMARTS, Swift, and other instruments), examine the cross-correlation between the light curves measured in the different energy bands, and interpret the resulting spectral energy distributions in the context of well-known blazar emission models. We find that the γ-ray activity is well correlated with a series of near-IR/optical flares, accompanied by an increase in the optical polarization degree. On the other hand, the X-ray light curve shows a distinct 20 day high state of unusually soft spectrum, which does not match the extrapolation of the optical/UV synchrotron spectrum. We tentatively interpret this feature as the bulk Compton emission by cold electrons contained in the jet, which requires an accretion disk corona with an effective covering factor of 19% at a distance of 100 R g. We model the broadband spectra with a leptonic model with external radiation dominated by the infrared emission from the dusty torus. © 2012. The American Astronomical Society. All rights reserved