Impact of contrast-enhanced ultrasound in patients with renal function impairment

To investigate the role of contrast enhanced ultrasound (CEUS) in evaluating patients with renal function impairment (RFI) showing: (1) acute renal failure (ARF) of suspicious vascular origin; or (2) suspicious renal lesions

DEVELOPMENT OF A LARGE EDDY SIMULATION MODEL FOR THE STUDY OF POLLUTANT DISPERSION IN URBAN AREAS

2008/2009In this thesis, a new large-eddy simulation solver, LES-AIR, has been developed, tested and applied to a practical situation of flow and pollutant dispersion in urban environments. The novelty of the present research resides in the application of a high resolution, accurate, CFD technique to the simulation of real-life flows. The code uses a body fitted curvilinear grid to account for the macro geometry such as terrain slopes, and is thus able to reproduce in detail the complex conditions typical of urban areas; by utilizing the technique of immersed boundaries, the code is also able to mimic the presence the micro complexities such as anthropic structures (i.e. buildings). The first part of the thesis presents a detailed description of the mathematical and numerical model on which the code is based. An extensive set of validation tests was performed in flow configurations having an increasing degree of complexity in terms of forcing and geometry. The numerical model thus validated is applied for obtaining flow and pollutant dispersion in the Servola-Valmaura suburban area of the city of Trieste in Italy. The pollutant was introduced into the domain from a line source near the ground, mimicking the emission from vehicular traffic. In spite of the idealizations inherent to the model, LES-AIR is able to predict the flow and dispersion patterns well, and has proven to be a reliable tool for adaptation in urban pollution studies.Nella presente tesi è stato sviluppato, testato ed applicato ad un caso studio applicativo un nuovo solutore numerico, chiamato codice LES-AIR, capace di predire i campi di vento e la dispersione di nquinanti in ambienti urbani. La maggiore novità di questo lavoro risiede nell’utilizzo di una tecnica fluidodinamica molto accurata e ad alta risoluzione per la simulazione di flussi reali. Il codice LES-AIR è capace di riprodurre con grande dettaglio le geometrie complesse tipiche delle aree urbane tramite l’utilizzo congiunto di una griglia curvilinea, che si adatta all’ orografia del terreno, e della tecnica dei corpi immersi, con la quale vengono riprodotti gli ostacoli antropici, quali gli edifici. Nella prima parte della tesi viene fornita una descrizione dettagliata del modello matematico e numerico su cui si basa il codice. Il modello è stato validato per mezzo di un esteso set di casi test, aventi un grado crescente di complessit à in termini di forzanti e di configurazione geometriche. Il modello così validato è stato applicato alla riproduzione di un caso applicativo nel quale i campi di vento e la dispersione di un inquinante nella zona di Servola-Valmaura, situata nella periferia di Trieste, sono stati simulati. L’ inquinante è stato introdotto da una sorgente lineare posta in prosimità del terreno e rappresentante l’emissione derivante dal traffico cittadino. Nonostante le condizioni idealizzate di vento considerate, il codice LES-AIR si è dimostrato molto efficace nella predizione del flusso e della dispersione dell’inquinante e quindi si è attestato essere un valido strumento negli studi d’ inquinamento urbani.XXII Ciclo198

Disease-associated missense mutations in GluN2B subunit alter NMDA receptor ligand binding and ion channel properties.

Genetic and bioinformatic analyses have identified missense mutations in GRIN2B encoding the NMDA receptor GluN2B subunit in autism, intellectual disability, Lennox Gastaut and West Syndromes. Here, we investigated several such mutations using a near-complete, hybrid 3D model of the human NMDAR and studied their consequences with kinetic modelling and electrophysiology. The mutants revealed reductions in glutamate potency; increased receptor desensitisation; and ablation of voltage-dependent Mg block. In addition, we provide new views on Mg and NMDA channel blocker binding sites. We demonstrate that these mutants have significant impact on excitatory transmission in developing neurons, revealing profound changes that could underlie their associated neurological disorders. Of note, the NMDAR channel mutant GluN2B unusually allowed Mg permeation, whereas nearby N615I reduced Ca permeability. By identifying the binding site for an NMDAR antagonist that is used in the clinic to rescue gain-of-function phenotypes, we show that drug binding may be modified by some GluN2B disease-causing mutations

CXCL12 inhibits expression of the NMDA receptor's NR2B subunit through a histone deacetylase-dependent pathway contributing to neuronal survival

Homeostatic chemokines, such as CXCL12, can affect neuronal activity by the regulation of inhibitory and excitatory neurotransmission, but the mechanisms involved are still undefined. Our previous studies have shown that CXCL12 protects cortical neurons from excitotoxicity by promoting the function of the gene-repressor protein Rb, which is involved in the recruitment of chromatin modifiers (such as histone deacetylases (HDACs)) to gene promoters. In neurons, Rb controls activity-dependent genes essential to neuronal plasticity and survival, such as the N-methyl--aspartic acid (NMDA) receptor's subunit NR2B, the expression of which in the tetrameric ion channel largely affects calcium signaling by glutamate. In this study, we report that CXCL12 differentially modulates intracellular responses after stimulation of synaptic and extrasynaptic NMDA receptors, by a specific regulation of the NR2B gene that involves HDACs. Our results show that CXCL12 selectively inhibits NR2B expression in vitro and in vivo altering NMDA-induced calcium responses associated with neuronal death, while promoting prosurvival pathways that depend on stimulation of synaptic receptors. Along with previous studies, these findings underline the role of CXCL12/CXCR4 in the regulation of crucial components of glutamatergic transmission. These novel effects of CXCL12 may be involved in the physiological function of the chemokine in both developing and mature brains

Reelin Secreted by GABAergic Neurons Regulates Glutamate Receptor Homeostasis

BACKGROUND: Reelin is a large secreted protein of the extracellular matrix that has been proposed to participate to the etiology of schizophrenia. During development, reelin is crucial for the correct cytoarchitecture of laminated brain structures and is produced by a subset of neurons named Cajal-Retzius. After birth, most of these cells degenerate and reelin expression persists in postnatal and adult brain. The phenotype of neurons that bind secreted reelin and whether the continuous secretion of reelin is required for physiological functions at postnatal stages remain unknown. METHODOLOGY/PRINCIPAL FINDINGS: Combining immunocytochemical and pharmacological approaches, we first report that two distinct patterns of reelin expression are present in cultured hippocampal neurons. We show that in hippocampal cultures, reelin is secreted by GABAergic neurons displaying an intense reelin immunoreactivity (IR). We demonstrate that secreted reelin binds to receptors of the lipoprotein family on neurons with a punctate reelin IR. Secondly, using calcium imaging techniques, we examined the physiological consequences of reelin secretion blockade. Blocking protein secretion rapidly and reversibly changes the subunit composition of N-methyl-D-aspartate glutamate receptors (NMDARs) to a predominance of NR2B-containing NMDARs. Addition of recombinant or endogenously secreted reelin rescues the effects of protein secretion blockade and reverts the fraction of NR2B-containing NMDARs to control levels. Therefore, the continuous secretion of reelin is necessary to control the subunit composition of NMDARs in hippocampal neurons. CONCLUSIONS/SIGNIFICANCE: Our data show that the heterogeneity of reelin immunoreactivity correlates with distinct functional populations: neurons synthesizing and secreting reelin and/or neurons binding reelin. Furthermore, we show that continuous reelin secretion is a strict requirement to maintain the composition of NMDARs. We propose that reelin is a trans-neuronal messenger secreted by GABAergic neurons that regulates NMDARs homeostasis in postnatal hippocampus. Defects in reelin secretion could play a major role in the development of neuropsychiatric disorders, particularly those associated with deregulation of NMDARs such as schizophrenia

Increased contribution of NR2A subunit to synaptic NMDA receptors in developing rat cortical neurons

Pharmacologically isolated miniature NMDA receptor-mediated excitatory postsynaptic currents (mN-EPSCs) were recorded in large visual cortical neurons in layer V of rat cortical slices. Haloperidol (100 μm) and CP101,606 (10 μm), two specific blockers of NMDA receptors comprising NR1/NR2B subunits, were tested on mN-EPSCs in rats at postnatal days 7 and 8 (P7–P8) and P13–P15. At both ages tested, no significant effects of these drugs were seen in the whole population of neurons, although in few neurons at both ages changes in amplitude were observed with haloperidol. Other dopamine receptor antagonists, spiperone and clozapine, failed to decrease mN-EPSCs in cortical neurons at P13–P15.CP101,606 (10 μm) significantly decreased the amplitude of evoked N-EPSCs (eN-EPSCs) in visual cortical slices from rats at P3–P5, a developmental stage at which mRNA studies have indicated the virtual absence of NR2A mRNA. CP101,606 failed to significantly change evoked AMPA-mediated EPSCs at P5 and eN-EPSCs at P7–P8 and P13–P15.NMDA receptor-mediated currents were also studied in somatic outside-out patches at P13–P15 with fast application of l-glutamate (1 mm). Haloperidol (50 μm) and CP101,606 (10 μm) blocked these currents in all patches tested. The effect of CP101,606 was concentration dependent.We suggest that rather early in development synaptic receptors comprising NR1/NR2B subunits could be associated with other subunits so that blockade by haloperidol and CP101,606 is prevented. Moreover, the consistent blockade seen in outside out patches might be ascribed to the confinement of NR1/NR2B receptors to an extrasynaptic population