Confronto fra invecchiamento accelerato e naturale di rivestimenti organici protettivi: uso di dati meteorologici

Confronto fra invecchiamento acceleratoe naturale di rivestimenti organiciprotettivi: uso di dati meteorologiciF. Deflorian, S. Rossi, L. Fedrizzi, C. ZanellaLe proprietà protettive dei rivestimenti organici per applicazioni per esterni sono tradizionalmentevalutate attraverso test accelerati di laboratorio. Una questione aperta rimane la reale correlazionefra questi test accelerati e l’esposizione naturale in ambienti diversi. In questo lavoro è descrittoun nuovo approccio basato sulla semplice idea di quantificare il grado di invecchiamento naturaleutilizzando un numero limitato di parametri ambientali che possono influenzare le proprietà protettive:l’energia totale nell’intervallo UV, il tempo di esposizione ad umidità relative superiori ad una certasoglia (tale da causare condensa superficiale), il tempo di esposizione ad una temperature superiorealla Tg del polimero, ecc. In questo lavoro si è cercato di applicare questo approccio a un rivestimentopoliestere ottenuto per coil-coating per applicazioni per esterni. I primi risultai sono molto interessantie incoraggiano ulteriori sviluppi che considerino un maggior numero di materiali e siti naturalidi esposizione

Valutazione della vita utile di acciaio zincato verniciato: esposizioni su campo e prove di laboratorio

Al fine di studiare a fondo le relazioni che intercorrono tra l’invecchiamento artificiale e quello naturale,sei diversi campioni zincati e rivestiti con primer e top coat sono stati esposti per 12 mesi in 3 diversi ambientinaturali: Trento (Italia), rappresentativo del clima continentale, Marsiglia (Francia) di clima mediterraneoe Daytona (USA) di ambiente tropicale umido. In tali siti sono stati raccolti, durante l’esposizione, i principalidati meteorologici. I rivestimenti sono stati testati in laboratorio mediante prove accelerate tradizionali(camera a nebbia salina, esposizione UV, etc). L’analisi delle proprietà protettive dei rivestimentie il monitoraggio dello stato di degrado è stato effettuato mediante misure elettrochimiche.I risultati dimostrano che il tempo trascorso in presenza di condensa e cloruri sembra il fattore determinanteil degrado del sistema di protezione. L'esposizione naturale induce un degrado misurabile, in gradodi classificare l'aggressività dei diversi siti di esposizione. Questo fatto è dovuto alla presenza di cloruri,alla maggiore energia di radiazione solare e al maggior tempo di condensa. L'ambiente urbano,con la presenza di inquinanti, può influenzare maggiormente il degrado del substrato metallico,più che influire sul deterioramento delle proprietà di barriera del rivestimento organico

Electrochemical properties and electrochemical impedance spectroscopy of polypyrrole-coated platinum electrodes

Polypyrrole (PPy) films of different thickness were characterized by electrochemical impedance spectroscopy (EIS) measurements in acetonitrile and aqueous solutions, containing 0.1 M NaClO4 or sodium dodecylsulfate as the dopant. The PPy films were electrochemically deposited on Pt, and their electrochemical properties studied by cyclic voltammetry. Impedance spectra were obtained at potentials ranging from 0 to 0.8 V/SCE. The EIS data were fitted using two different equivalent electrical circuits (depending on the nature of the dopant). They involve a diffusive capacitance, which increased with the passing charge during electrosynthesis (i.e. film thickness) for ClO4--doped PPy, but was practically unaffected by the film thickness in the case of SDS-doped PPy. Also, a double-layer capacitance was found only in the circuit of ClO4--doped PPy. It increased with the film thickness, and showed a minimum near the open-circuit potential. Finally the charge-transfer resistance (Rct) obtained with SDS is nearly 200-fold higher than that obtained with ClO4- in the same solvent (H2O). With the same dopant (ClO4-), Rct is about five times higher in acetonitrile relative to water. All these EIS results of the different types of PPy suggest a relation with the wettability of the polymer.KEY WORDS: Conducting polymers, Polypyrrole, Electrochemical impedance spectroscopy, Equivalent-electrical circuit, Micellar mediaBull. Chem. Soc. Ethiop. 2006, 20(2), 279-293

EFFETTO DI INCLUSIONI METALLICHE NEI PROCESSI DI ANODIZZAZIONE DI LEGHE DI ALLUMINIO

La necessità di lavorare mediante asportazione di truciolo leghe di alluminio ha da tempo portato allo sviluppo di leghe specifiche, far cui le più diffuse contengono piombo. Tali leghe devono ora essere superate a causa delle restrizioni legate alla presenza del piombo, motivate principalmente da ragioni ambientali. Diverse leghe sono state sviluppate cercando di sostituire il piombo con il manganese, il ferro, il nichel o l’indio. Un altro promettente sostituto del piombo per queste applicazioni è il berillio, oggetto di studio negli ultimi anni come alligante dell’alluminio. Molto spesso per applicazioni che spaziano dalla motoristica alla attrezzatura sportiva alla componentistica per l’elettronica, i componenti in lega di alluminio devono essere modificati superficialmente per migliorarne sia la resistenza alla corrosione che le proprietà tribologiche. Uno dei trattamenti più diffusi è l’anodizzazione. La presenza però di una particolare microstruttura, contenente isole di metallo elettrochimicamente più nobile (Pb o Bi) può notevolmente influenzare la qualità dell’ossido anodico e di conseguenza le proprietà di resistenza alla corrosione del componente anodizzato. Tale aspetto non è stato ancora sufficientemente analizzato scientificamente. In questo studio sono stati considerati campioni di diverse leghe di alluminio contenenti sia Pb che Bi, sottoposti a trattamenti di anodizzazione. Sono state studiate le difettosità riscontrate nell’ossido individuate mediante microscopia elettronica ed ottica, collegandole al comportamento corrosivo e alla microstruttura del materiale (dimensioni e distribuzione delle isole di Pb o di Bi). Sono state eseguite prove elettrochimiche di vario tipo al fine di comprendere il meccanismo di degrado. Lo scopo finale del lavoro sperimentale è quello di fornire informazioni utili all’ottimizzazione di nuove leghe di alluminio, lavorabili per asportazione di truciolo esenti piombo, che mantengano buone proprietà protettive dell’ossido anodico

Impact of protein-ligand solvation and desolvation on transition state thermodynamic properties of adenosine A2Aligand binding kinetics

Ligand-protein binding kinetic rates are growing in importance as parameters to consider in drug discovery and lead optimization. In this study we analysed using surface plasmon resonance (SPR) the transition state (TS) properties of a set of six adenosine A2Areceptor inhibitors, belonging to both the xanthine and the triazolo-triazine scaffolds. SPR highlighted interesting differences among the ligands in the enthalpic and entropic components of the TS energy barriers for the binding and unbinding events. To better understand at a molecular level these differences, we developed suMetaD, a novel molecular dynamics (MD)-based approach combining supervised MD and metadynamics. This method allows simulation of the ligand unbinding and binding events. It also provides the system conformation corresponding to the highest energy barrier the ligand is required to overcome to reach the final state. For the six ligands evaluated in this study their TS thermodynamic properties were linked in particular to the role of water molecules in solvating/desolvating the pocket and the small molecules. suMetaD identified kinetic bottleneck conformations near the bound state position or in the vestibule area. In the first case the barrier is mainly enthalpic, requiring the breaking of strong interactions with the protein. In the vestibule TS location the kinetic bottleneck is instead mainly of entropic nature, linked to the solvent behaviour

IRSp53 controls plasma membrane shape and polarized transport at the nascent lumen in epithelial tubules

It is unclear whether the establishment of apical\u2013basal cell polarity during the generation of epithelial lumens requires molecules acting at the plasma membrane/actin interface. Here, we show that the I-BAR-containing IRSp53 protein controls lumen formation and the positioning of the polarity determinants aPKC and podocalyxin. Molecularly, IRSp53 acts by regulating the localization and activity of the small GTPase RAB35, and by interacting with the actin capping protein EPS8. Using correlative light and electron microscopy, we further show that IRSp53 ensures the shape and continuity of the opposing plasma membrane of two daughter cells, leading to the formation of a single apical lumen. Genetic removal of IRSp53 results in abnormal renal tubulogenesis, with altered tubular polarity and architectural organization. Thus, IRSp53 acts as a membrane curvature-sensing platform for the assembly of multi-protein complexes that control the trafficking of apical determinants and the integrity of the luminal plasma membrane

IRSp53 controls plasma membrane shape and polarized transport at the nascent lumen in epithelial tubules

It is unclear whether the establishment of apical–basal cell polarity during the generation of epithelial lumens requires molecules acting at the plasma membrane/actin interface. Here, we show that the I-BAR-containing IRSp53 protein controls lumen formation and the positioning of the polarity determinants aPKC and podocalyxin. Molecularly, IRSp53 acts by regulating the localization and activity of the small GTPase RAB35, and by interacting with the actin capping protein EPS8. Using correlative light and electron microscopy, we further show that IRSp53 ensures the shape and continuity of the opposing plasma membrane of two daughter cells, leading to the formation of a single apical lumen. Genetic removal of IRSp53 results in abnormal renal tubulogenesis, with altered tubular polarity and architectural organization. Thus, IRSp53 acts as a membrane curvature-sensing platform for the assembly of multi-protein complexes that control the trafficking of apical determinants and the integrity of the luminal plasma membrane

Characterization of the Regulatory Region of the Zebrafish Prep1.1 Gene: Analogies to the Promoter of the Human PREP1

Prep1 is a developmentally essential TALE class homeodomain transcription factor. In zebrafish and mouse, Prep1 is already ubiquitously expressed at the earliest stages of development, with important tissue-specific peculiarities. The Prep1 gene in mouse is developmentally essential and has haploinsufficient tumor suppressor activity [1]. We have determined the human Prep1 transcription start site (TSS) by primer extension analysis and identified, within 20 bp, the transcription start region (TSR) of the zebrafish Prep1.1 promoter. The functions of the zebrafish 5′ upstream sequences were analyzed both by transient transfections in Hela Cells and by injection in zebrafish embryos. This analysis revealed a complex promoter with regulatory sequences extending up to −1.8, possibly −5.0 Kb, responsible for tissue specific expression. Moreover, the first intron contains a conserved tissue-specific enhancer both in zebrafish and in human cells. Finally, a two nucleotides mutation of an EGR-1 site, conserved in all species including human and zebrafish and located at a short distance from the TSS, destroyed the promoter activity of the −5.0 Kb promoter. A transgenic fish expressing GFP under the −1.8 Kb zebrafish promoter/enhancer co-expressed GFP and endogenous Prep1.1 during embryonic development. In the adult fish, GFP was expressed in hematopoietic regions like the kidney, in agreement with the essential function of Prep1 in mouse hematopoiesis. Sequence comparison showed conservation from man to fish of the sequences around the TSS, within the first intron enhancer. Moreover, about 40% of the sequences spread throughout the 5 Kbof the zebrafish promoter are concentrated in the −3 to −5 Kb of the human upstream region

Dynamic phosphorylation of Histone Deacetylase 1 by Aurora kinases during mitosis regulates zebrafish embryos development

Histone deacetylases (HDACs) catalyze the removal of acetyl molecules from histone and nonhistone substrates playing important roles in chromatin remodeling and control of gene expression. Class I HDAC1 is a critical regulator of cell cycle progression, cellular proliferation and differentiation during development; it is also regulated by many post-translational modifications (PTMs). Herein we characterize a new mitosis-specific phosphorylation of HDAC1 driven by Aurora kinases A and B. We show that this phosphorylation affects HDAC1 enzymatic activity and it is critical for the maintenance of a proper proliferative and developmental plan in a complex organism. Notably, we find that Aurora-dependent phosphorylation of HDAC1 regulates histone acetylation by modulating the expression of genes directly involved in the developing zebrafish central nervous system. Our data represent a step towards the comprehension of HDAC1 regulation by its PTM code, with important implications in unravelling its roles both in physiology and pathology

X‐Ray Crystallography and Free Energy Calculations Reveal the Binding Mechanism of A2A Adenosine Receptor Antagonists

We present a robust protocol based on iterations of free energy perturbation (FEP) calculations, chemical synthesis, biophysical mapping and X‐ray crystallography to reveal the binding mode of an antagonist series to the A2A adenosine receptor (AR). Eight A2AAR binding site mutations from biophysical mapping experiments were initially analyzed with sidechain FEP simulations, performed on alternate binding modes. The results distinctively supported one binding mode, which was subsequently used to design new chromone derivatives. Their affinities for the A2AAR were experimentally determined and investigated through a cycle of ligand‐FEP calculations, validating the binding orientation of the different chemical substituents proposed. Subsequent X‐ray crystallography of the A2AAR with a low and a high affinity chromone derivative confirmed the predicted binding orientation. The new molecules and structures here reported were driven by free energy calculations, and provide new insights on antagonist binding to the A2AAR, an emerging target in immuno‐oncologyThis work was financially supported by the Swedish Research Council (Grant 521‐2014‐2118); Consellería de Cultura, Educación e Ordenación Universitaria of the Galician Government (Grant ED431B2017/70); Centro Singular de Investigación de Galicia accreditation 2016–2019 (Grant ED431G/09), and the European Regional Development Fund (ERDF). Additional support from the Swedish strategic research program eSSENCE is acknowledged. The computations were performed on resources provided by the Swedish National Infrastructure for Computing (SNIC). This research program has been developed in the frame of the European COST action ERNEST (Grant CA 18133) and GLISTEN (Grant CA 1207)S