SDEC Audit. Progetto per la verifica applicativa dello Schema di Sviluppo dello Spazio Europeo attraverso reti interistituzionali nell'ambito di Interreg III. Rapporto finale

Rapporto operativo per il Ministero dei lavori pubblic

ESPON Project 2.3.1., Application and effects of the ESDP in Member States. First Interim Report

This First Interim Report includes the initial results of the project “Application and Effects of the ESDP in the Member States” within the ESPON Programme 2000-2006. The focus of the study is the application of the European Spatial Development Perspective (ESDP), which was adopted at the Potsdam European Council meeting in May 1999

Calcineurin controls expression of EAAT1/GLAST in mouse and human cultured astrocytes through dynamic regulation of protein synthesis and degradation

Alterations in the expression of glutamate/aspartate transporter (GLAST) have been associated with several neuropathological conditions including Alzheimer\u2019s disease and epilepsy. However, the mechanisms by which GLAST expression is altered are poorly understood. Here we used a combination of pharmacological and genetic approaches coupled with quantitative PCR and Western blot to investigate the mechanism of the regulation of GLAST expression by a Ca2+ /calmodulin-activated phosphatase calcineurin (CaN). We show that treatment of cultured hippocampal mouse and fetal human astrocytes with a CaN inhibitor FK506 resulted in a dynamic modulation of GLAST protein expression, being downregulated after 24\u201348 h, but upregulated after 7 days of continuous FK506 (200 nM) treatment. Protein synthesis, as assessed by puromycin incorporation in neo-synthesized polypeptides, was inhibited already after 1 h of FK506 treatment, while the use of a proteasome inhibitor MG132 (1 \ub5M) shows that GLAST protein degradation was only suppressed after 7 days of FK506 treatment. In astrocytes with constitutive genetic ablation of CaN both protein synthesis and degradation were significantly inhibited. Taken together, our data suggest that, in cultured astrocytes, CaN controls GLAST expression at a posttranscriptional level through regulation of GLAST protein synthesis and degradation

Quantification of the Chemical Chaperone 4-Phenylbutyric Acid (4-PBA) in Cell Culture Media via LC-HRMS: Applications in Fields of Neurodegeneration and Cancer

In recent years, 4-phenylbutyric acid (4-PBA), an FDA-approved drug, has increasingly been used as a nonspecific chemical chaperone in vitro and in vitro, but its pharmacodynamics is still not clear. In this context, we developed and validated a Liquid Chromatography–High Resolution Mass Spectrometry (LC-HRMS) method to quantify 4-PBA in NeuroBasal-A and Dulbecco’s Modified Eagle widely used cell culture media. Samples were injected on a Luna® 3 µm PFP(2) 100 Å (100 × 2.0 mm) column maintained at 40 °C. Water and methanol both with 0.1% formic acid served as mobile phases in a step gradient mode. The mass acquisition was performed by selected ion monitoring (SIM) in negative mode for a total run time of 10.5 min at a flow rate of 0.300 mL/min. The analogue 4-(4-Nitrophenyl)-Butyric Acid served as internal standard. Validation parameters were verified according to FDA and EMA guidelines. The quantification ranges from 0.38–24 µM. Inter and intraday RSDs (Relative Standard Deviations) were within 15%. The developed LC-HRMS method allowed the estimation of 4-PBA absorption and adsorption kinetics in vitro in two experimental systems: (i) 4-PBA improvement of protein synthesis in an Alzheimer’s disease astrocytic cell model; and (ii) 4-PBA reduction of endoplasmic reticulum stress in thapsigargin-treated melanoma cell lines. © 2023 by the authors

Equilibrium and nonequilibrium description of negative temperature states in a one-dimensional lattice using a wave kinetic approach

We predict negative temperature states in the discrete nonlinear Schödinger (DNLS) equation as exact solutions of the associated wave kinetic equation. Within the wave kinetic approach, we define an entropy that results monotonic in time and reaches a stationary state, that is consistent with classical equilibrium statistical mechanics. We also perform a detailed analysis of the fluctuations of the actions at fixed wave numbers around their mean values. We give evidence that such fluctuations relax to their equilibrium behavior on a shorter timescale than the one needed for the spectrum to reach the equilibrium state. Numerical simulations of the DNLS equation are shown to be in agreement with our theoretical results. The key ingredient for observing negative temperatures in lattices characterized by two invariants is the boundedness of the dispersion relation

Modelling Discharge Rates and Ground Settlement Induced by Tunnel Excavation

Interception of aquifers by tunnel excavation results in water inflow and leads to drawdown of the water table which may induce ground settlement. In this work, analytical and numerical models are presented which specifically address these groundwater related processes in tunnel excavation. These developed models are compared and their performance as predictive tools is evaluated. Firstly, the water inflow in deep tunnels is treated. It is shown that introducing a reduction factor accounting for the effect of effective stress on hydrodynamic parameters avoids overestimation. This effect can be considered in numerical models using effective stress-dependent parameters. Then, quantification of ground settlement is addressed by a transient analytical solution. These solutions are then successfully applied to the data obtained during the excavation of the La Praz exploratory tunnel in the Western Alps (France), validating their usefulness as predictive tools