Zn(II)-curc targets p53 in thyroid cancer cells

P53 mutation is a common event in many cancers, including thyroid carcinoma. Defective p53 activity promotes cancer resistance to therapies and a more malignant phenotype, acquiring oncogenic functions. Rescuing the function of mutant p53 (mutp53) protein is an attractive anticancer therapeutic strategy. Zn(II)-curc is a novel small molecule that has been shown to target mutp53 protein in several cancer cells, but its effect in thyroid cancer cells remains unclear. Here, we investigated whether Zn(II)-curc could affect p53 in thyroid cancer cells with both p53 mutation (R273H) and wild-type p53. Zn(II)-curc induced mutp53H273 downregulation and reactivation of wild-type functions, such as binding to canonical target promoters and target gene transactivation. This latter effect was similar to that induced by PRIMA-1. In addition, Zn(II)-curc triggered p53 target gene expression in wild-type p53-carrying cells. In combination treatments, Zn(II)-curc enhanced the antitumor activity of chemotherapeutic drugs, in both mutant and wild-type-carrying cancer cells. Taken together, our data indicate that Zn(II)-curc promotes the reactivation of p53 in thyroid cancer cells, providing in vitro evidence for a potential therapeutic approach in thyroid cancers

Experimental investigation of the productivity of a wet separation process of traditional and bio-plastics

The separation process within a mechanical recycling plant plays a major role in the context of the production of high-quality secondary raw materials and the reduction of extensive waste disposal in landfills. Traditional plants for plastic separation employ dry or wet processes that rely on the different physical properties among the polymers. The hydraulic separator is a device employing a wet technology for particle separation. It allows the separation of two-polymer mixtures into two products, one collected within the instrument and the other one expelled through its outlet ducts. Apparatus performance were analyzed as a function of fluid and solid flow rates, flow patterns developing within the apparatus, in addition to the density, shape, and size of the polymers. For the hydraulic configurations tested, a two-way coupling takes place where the fluid exerts an influence on the plastic particles and the opposite occurs too. The interaction between the solid and liquid phases determines whether a certain polymer settles within the device or is expelled from the apparatus. Tests carried out with samples of increasing volumes of solid particles demonstrate that there are no significant differences in the apparatus effectiveness as far as a two-way interaction takes place. Almost pure concentrates of Polyethylene Terephthalate (PET), Polyvinyl Chloride (PVC), and Polycarbonate (PC) can be obtained from a mixture of traditional polymers. Tests conducted on Polylactic Acid (PLA) and Mater-Bi® samples showed that the hydraulic separator can be effectively employed to separate bio-plastics from conventional plastics with remarkable grade and recovery

Transitions in auditory rehabilitation with bone conductive implant (bci)

Background: The bone conductive implants (BCI) are nowadays a reliable alternative for rehabilitation of specific forms of hearing loss, i.e. conductive, mixed or single sided deafness (SSD). Aims/Objective: To analyse the various factors in play when considering an auditory rehabilitation with a bone-conductive device (BCI). Materials and Methods: The clinical charts of subjects who underwent BCI application at the same Implanting Center from 2005 to 2018 were retrieved analysing also the reason for eventual explantation and the alternative option (transition) for hearing rehabilitation. Results: Nine BAHA Compact, 4 BAHA Intenso, 21 BAHA Divino, 3 BAHA BP100, 4 Ponto, 2 Sophono, 5 Bonebridge, 5 BAHA5 Attract; 11 BAHA5 Connect were used in 12 unilateral COM; 16 bilateral COM; 3 unilateral cholesteatoma; 6 bilateral cholesteatoma; 2 unilateral otosclerosis; 5 bilateral otosclerosis; 9 congenital malformations; 6 major otoneurosurgical procedures; 5 sudden deafness. Explantation was necessary for five subjects. Conclusions: Middle ear pathology and sequels from surgery represent the most common reason for BCI implantation, both in unilateral and in bilateral cases. Transition from one implantable device to another one can be predictable, mostly when explantation is necessary. Significance: The role of BCI for rehabilitation in middle ear pathology may be extremely important

Branched-chain amino acids, mitochondrial biogenesis, and healthspan: an evolutionary perspective

Malnutrition is common among older persons, with important consequences increasing frailty and morbidity and reducing health expectancy. On the contrary, calorie restriction (CR, a low-calorie dietary regimen with adequate nutrition) slows the progression of age-related diseases and extends the lifespan of many species. Identification of strategies mimicking key CR mechanisms – increased mitochondrial respiration and reduced production of oxygen radicals – is a hot topic in gerontology. Dietary supplementation with essential and/or branched chain amino acids (BCAAs) exerts a variety of beneficial effects in experimental animals and humans and has been recently demonstrated to support cardiac and skeletal muscle mitochondrial biogenesis, prevent oxidative damage, and enhance physical endurance in middle-aged mice, resulting in prolonged survival. Here we review recent studies addressing the possible role of BCAAs in energy metabolism and in the longevity of species ranging from unicellular organisms to mammals. We also summarize observations from human studies supporting the exciting hypothesis that dietary BCAA enriched mixture supplementation might be a health-promoting strategy in aged patients at risk

Unveiling the folding mechanism of the bromodomains

Bromodomains (BRDs) are small protein domains often present in large multidomain proteins involved in transcriptional regulation in eukaryotic cells. They currently represent valuable targets for the development of inhibitors of aberrant transcriptional processes in a variety of human diseases. Here we report urea-induced equilibrium unfolding experiments monitored by circular dichroism (CD) and fluorescence on two structurally similar BRDs: BRD2(2) and BRD4(1), showing that BRD4(1) is more stable than BRD2(2). Moreover, we report a description of their kinetic folding mechanism, as obtained by careful analysis of stopped-flow and temperature-jump data. The presence of a high energy intermediate for both proteins, suggested by the non-linear dependence of the folding rate on denaturant concentration in the millisec time regime, has been experimentally observed by temperature-jump experiments. Quantitative global analysis of all the rate constants obtained over a wide range of urea concentrations, allowed us to propose a common, three-state, folding mechanism for these two BRDs. Interestingly, the intermediate of BRD4(1) appears to be more stable and structurally native-like than that populated by BRD2(2). Our results underscore the role played by structural topology and sequence in determining and tuning the folding mechanism

Repurposing designed mutants: a valuable strategy for computer-aided laccase engineering – the case of POXA1b

The broad specificity of laccases, a direct consequence of their shallow binding site, makes this class of enzymes a suitable template to build specificity toward putative substrates. In this work, a computational methodology that accumulates beneficial interactions between the enzyme and the substrate in productive conformations is applied to oxidize 2,4-diamino-benzenesulfonic acid with POXA1b laccase. Although the experimental validation of two designed variants yielded negative results, most likely due to the hard oxidizability of the target substrate, molecular simulations suggest that a novel polar binding scaffold was designed to anchor negatively charged groups. Consequently, the oxidation of three such molecules, selected as representative of different classes of substances with different industrial applications, significantly improved. According to molecular simulations, the reason behind such an improvement lies in the more productive enzyme–substrate binding achieved thanks to the designed polar scaffold. In the future, mutant repurposing toward other substrates could be first carried out computationally, as done here, testing molecules that share some similarity with the initial target. In this way, repurposing would not be a mere safety net (as it is in the laboratory and as it was here) but rather a powerful approach to transform laccases into more efficient multitasking enzymes.This work was funded by INDOX (KBBE-2013-7-613549) European project and CTQ2013-48287-R Spanish National Project. V. G. and E. M. acknowledge Università degli Studi di Napoli and Generalitat de Catalunya for their respective predoctoral fellowships.Peer ReviewedPostprint (author's final draft

Effect of bet missense mutations on bromodomain function, inhibitor binding and stability

Lysine acetylation is an important epigenetic mark regulating gene transcription and chromatin structure. Acetylated lysine residues are specifically recognized by bromodomains, small protein interaction modules that read these modification in a sequence and acetylation dependent way regulating the recruitment of transcriptional regulators and chromatin remodelling enzymes to acetylated sites in chromatin. Recent studies revealed that bromodomains are highly druggable protein interaction domains resulting in the development of a large number of bromodomain inhibitors. BET bromodomain inhibitors received a lot of attention in the oncology field resulting in the rapid translation of early BET bromodomain inhibitors into clinical studies. Here we investigated the effects of mutations present as polymorphism or found in cancer on BET bromodomain function and stability and the influence of these mutants on inhibitor binding. We found that most BET missense mutations localize to peripheral residues in the two terminal helices. Crystal structures showed that the three dimensional structure is not compromised by these mutations but mutations located in close proximity to the acetyl-lysine binding site modulate acetyl-lysine and inhibitor binding. Most mutations affect significantly protein stability and tertiary structure in solution, suggesting new interactions and an alternative network of protein-protein interconnection as a consequence of single amino acid substitution. To our knowledge this is the first report studying the effect of mutations on bromodomain function and inhibitor binding

Valutazione mediante tecnica fotografica dell'oscuramento della radiazione solare dovuto alle ostruzioni

Si propone un nuovo metodo che consente di utilizzare l’immagine fotografica di ostacoli ombreggianti per valutarne l’effetto oscurante nei confronti della radiazione solare diretta incidente su di una superficie. Il metodo proposto è simile a quello che prevede l’impiego del diagramma cilindrico dei percorsi solari, calcolati in funzione dell’azimuth e dell’altezza solare, e la sovrapposizione su di esso del poligono raffigurante l’oggetto ostruente costruito in base alle coordinate cilindriche dei suoi punti più significativi. La nuova procedura prevede invece la sovrapposizione sulla fotografia del percorso solare opportunamente trasformato secondo le regole della prospettiva. E’ così possibile tracciare direttamente sulla foto la posizione assunta dal sole agli orari richiesti per qualunque giorno dell’anno e località, utilizzando a tale scopo la conoscenza di un numero estremamente limitato di dati, peraltro facilmente reperibili. Le porzioni del percorso intercettate dal contorno degli ostacoli rappresentati nell’immagine fotografica indicano le fasce orarie durante le quali il punto della superficie ricevente risulta ombreggiato. Viceversa i restanti tratti del percorso solare evidenziano la presenza della componente diretta radiazione. Sono presentati, inoltre, i risultati del confronto tra i dati calcolati e quelli sperimentali condotti presso il Dipartimento di Ricerche Energetiche e Ambientali (DREAM) di Palermo