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

Characterization of a New Zinc Fixed-Point Cell for ITS-90 Realization

A new zinc fixed-point cell for the dissemination of International Temperature Scale (ITS-90) was realized at the Italian National Metrological Institute (Istituto Nazionale di Ricerca Metrologica, INRiM). This paper presents the results of its characterization, including fabrication details. In particular, immersion effects and influences of impurities on the freezing point of zinc were studied. The new open-type cell was prepared using a high purity sample, chemically analyzed, and the depression of the fixed-point temperature was calculated using the method of Sum of Individual Estimates (SIE). The new cell presents a smaller freezing point depression compared with the national reference for which the Overall Maximum Estimate (OME) method was applied. This behavior was confirmed also by the direct comparison of the two cells. These results provide confidence on the agreement between the experimental comparison and the SIE/OME evaluation. Finally, the improvement of the new zinc cell is reflected also in a lower uncertainty budget for the fixed-point realization

Realisation of the triple-point of Argon: comparison between two devices

A new cryostat for the realization of the triple-point of the argon (83.8058 K), a defining fixed point of the International Temperature Scale of 1990 (ITS-90), was acquired at Italian National Metrological Institute (INRiM). The new system, manufactured by Fluke, is intended to substitute the current National reference, a model developed at BNM-INM in the 1975. The main difference between the two system is in the way to control the temperature. In the BNM-INM device the temperature is controlled adjusting the pressure of liquid nitrogen bath, in the Fluke system instead, an electrical heater wrapped around the argon cell is used, following cryogenic practice. This paper describes the result of the direct comparison and shows typical phase transitions obtained with the two argon systems. Then, a complete uncertainty budget is evaluated for the new Fluke system and compared with the National standard

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