Strategies against nonsense: oxadiazoles as translational readthrough-inducing drugs (TRIDs)

This review focuses on the use of oxadiazoles as translational readthrough-inducing drugs (TRIDs) to rescue the functional full-length protein expression in mendelian genetic diseases caused by nonsense mutations. These mutations in specific genes generate premature termination codons (PTCs) responsible for the translation of truncated proteins. After a brief introduction on nonsense mutations and their pathological effects, the features of various classes of TRIDs will be described discussing differences or similarities in their mechanisms of action. Strategies to correct the PTCs will be presented, particularly focusing on a new class of Ataluren-like oxadiazole derivatives in comparison to aminoglycosides. Additionally, recent results on the efficiency of new candidate TRIDs in restoring the production of the cystic fibrosis transmembrane regulator (CFTR) protein will be presented. Finally, a prospectus on complementary strategies to enhance the effect of TRIDs will be illustrated together with a conclusive paragraph about perspectives, opportunities, and caveats in developing small molecules as TRIDs

In vitro and in silico studies of polycondensed diazine systems as anti-parasitic agents

Parasitic diseases caused by protozoarian agents are still relevant today more than ever. Recently, we synthesized several polycondensed diazine derivatives by means 1,3-dipolar cycloaddition reactions. A broad selection of these compounds were submitted to in vitro biological screening against Plasmodium falciparum, Leishmania infantum, Trypanosoma brucei, and Trypanosoma cruzi, resulting active at micromolar level. Induced Fit Docking/MM-GBSA studies were performed giving interesting indications about the probable mechanism of action of the most active compound

In Silico Design, Synthesis and Biological Evaluation of Anticancer Arylsulfonamide Endowed with Anti-Telomerase Activity

Telomerase, a reverse transcriptase enzyme involved in DNA synthesis, has a tangible role in tumor progression. Several studies have evidenced telomerase as a promising target for developing cancer therapeutics. The main reason is due to the overexpression of telomerase in cancer cells (85–90%) compared with normal cells where it is almost unexpressed. In this paper, we used a structure-based approach to design potential inhibitors of the telomerase active site. The MYSHAPE (Molecular dYnamics SHared PharmacophorE) approach and docking were used to screen an in-house library of 126 arylsulfonamide derivatives. Promising compounds were synthesized using classical and green methods. Compound 2C revealed an interesting IC50 (33 ± 4 µM) against the K-562 cell line compared with the known telomerase inhibitor BIBR1532 IC50 (208 ± 11 µM) with an SI ~10 compared to the BALB/3-T3 cell line. A 100 ns MD simulation of 2C in the telomerase active site evidenced Phe494 as the key residue as well as in BIBR1532. Each moiety of compound 2C was involved in key interactions with some residues of the active site: Arg557, Ile550, and Gly553. Compound 2C, as an arylsulfonamide derivative, is an interesting hit compound that deserves further investigation in terms of optimization of its structure to obtain more active telomerase inhibitors

Evaluation of the IKKβ Binding of Indicaxanthin by Induced-Fit Docking, Binding Pose Metadynamics, and Molecular Dynamics

Background: Indicaxanthin, a betaxanthin belonging to the betalain class of compounds, has been recently demonstrated to exert significant antiproliferative effects inducing apoptosis of human melanoma cells through the inhibition of NF-κB as the predominant pathway. Specifically, Indicaxanthin inhibited IκBα degradation in A375 cells. In resting cells, NF-κB is arrested in the cytoplasm by binding to its inhibitor protein IκBα. Upon stimulation, IκBα is phosphorylated by the IKK complex, and degraded by the proteasome, liberating free NF-κB into the nucleus to initiate target gene transcription. Inhibition of the IKK complex leads to the arrest of the NF-κB pathway. Methods: To acquire details at the molecular level of Indicaxanthin’s inhibitory activity against hIKKβ, molecular modeling and simulation techniques including induced-fit docking (IFD), binding pose metadynamics (BPMD), molecular dynamics simulations, and MM-GBSA (molecular mechanics-generalized Born surface area continuum solvation) have been performed. Results: The computational calculations performed on the active and inactive form, and the allosteric binding site of hIKKβ, revealed that Indicaxanthin inhibits prevalently the active form of the hIKKβ. MM-GBSA computations provide further evidence of Indicaxanthin’s stability inside the active binding pocket with a binding free energy of −22.2 ± 4.3 kcal/mol with respect to the inactive binding pocket with a binding free energy of −20.7 ± 4.7 kcal/mol. BPMD and MD simulation revealed that Indicaxanthin is likely not an allosteric inhibitor of hIKKβ. Conclusion: As a whole, these in silico pieces of evidence show that Indicaxanthin can inhibit the active form of the hIKKβ adding novel mechanistic insights on its recently discovered ability to impair NF-κB signaling in melanoma A375 cells. Moreover, our results suggest the phytochemical as a new lead compound for novel, more potent IKKβ inhibitors to be employed in the treatment of cancer and inflammation-related conditions

Ionospheric Observatory Development At Mario Zucchelli Station

Since 1995 Italian Ionospheric Antarctic Observatory at Terra Nova Bay, now “MARIO ZUCCHELLI”, station (geographic coordinates: 74.70°S, 164.11°E) performs continuous and systematic ionospheric vertical soundings. Long time series of continuous and accurate ionospheric observations (more than one solar cycle) are necessary for a deeper understanding of the complex phenomena occurring in the upper atmosphere at high latitude; furthermore high rate soundings (at least four soundings per hour or more) contribute to the short-time prediction of the radiopropagation conditions and to the Space Weather. During 2003–2004 Antarctic campaign a new digital ionosonde, recently developed at the Istituto Nazionale di Geofisica e Vulcanologia (INGV) in Rome, (Italy), has been installed the Ionospheric Observatory and preliminary tests have been carried out. This new Advanced Ionospheric Sounder-INGV, briefly AIS, is integrated in a stand alone system during winter time: the sounding, device settings and data sending to Rome are completely automatic and remote programmable. Ionograms are available on line at the INGV web and ftp server. The new features of the Ionospheric Observatory are presented and preliminary statistics on the reliability and validation of the experimental observation are shown and discussed

IN THE SEARCH OF LEPTIN AGONISTS AS ANTI-OBESITY DRUGS: PROTEIN/PROTEIN DOCKING, MOLECULAR DYNAMICS, AND VIRTUAL SCREENING

The body weight control is a mechanism thinly regulated by several hormonal, metabolic, and nervous pathways (1). Recessive homozygous mutations in the ob/ob and db/db mouse strain cause extreme obesity. The products of the ob and db genes are leptin and its receptor, respectively. The leptin receptor is crucial for energy homeostasis and regulation of food uptake. Leptin is a 16 kDa hormone that is mainly secreted by fat cells into the bloodstream. Under normal circumstances, circulating leptin levels are proportionate to the fat body mass. Sensing of elevated leptin levels by the hypothalamic neurocircutry activates a negative feedback loop resulting in reduced food intake and increased energy expenditure. Decreased leptin concentrations lead to opposite effectsTherefore rational design of leptin agonists could be an appealing challenge in the battle against obesity. Unfortunately only the crystal structure of leptin is available, but not that of the leptin receptor. In this work, first, we built, by homology modelling, the leptin receptor starting from FASTA sequence and the similarity search of templates. The obtained model was used to perform a protein-protein docking with the crystal structure of leptine by means Gramm-X server, with the aim to define the complementary surfaces of the two proteins. The complex of leptin/leptin receptor was then used as starting point to carry out molecular dynamics simulations in water solvent to characterize the key residues involved into the protein-protein interaction. Snapshots of leptin were used as template to build a pharmacophore hypothesis to carry out virtual screening on a large database of compounds. (1) Friedman, J. M., Halaas, J. L. Nature 1998, 395, 763-770 IN THE SEARCH OF LEPTIN AGONISTS AS ANTI-OBESITY DRUGS: PROTEIN/PROTEIN DOCKING, MOLECULAR DYNAMICS, AND VIRTUAL SCREENIN

New low power pulse compressed ionosonde at Gibilmanna Ionospheric Observatory

A digital low power pulse compressed ionosonde was developed at the Istituto Nazionale di Geofisica e Vulcanologia (INGV), Rome, Italy. The aim of this Advanced Ionospheric Sounder, AIS-INGV, is to reduce the transmitted power and, consequently, weight, size, power consumption and hardware complexity. To compensate the power reduction the most advanced HF radar techniques such as the pulse compression and a phase coherent integration are used. The ionosonde is completely programmable and a PC supports the data acquisition, control, storage and on-line processing. The first prototype was installed at Gibilmanna Ionospheric Observatory (Sicily), an interesting location in the center of Mediterranean area. The new ionosonde will contribute to ionospheric database and real time knowledge of South European ionospheric conditions for space weather applications. In this work the first results (ionograms and autoscaled characteristics) are presented and briefly discussed

Importance of a real-time monitoring of the Earth's ionosphere

The ionosphere affects the electromagnetic wave propagation and then its study is important for Earth-Earth, satellite-Earth, and satellite-satellite communication purposes. Diffractive and refractive processes due to irregular electron density structures cause signal fluctuations that can disrupt satellite-ground communications and represent a hazard for navigation systems. The study and the real-time monitoring of the ionosphere are important for Space Weather purposes. The ionospheric vertical sounding is described, together with the automatic scaling of the ionograms.UnpublishedRome2A. Fisica dell'alta atmosferaope

SUsceptibility and Resistance to Fosfomycin and other antimicrobial agents among pathogens causing lower urinary tract infections: findings of the SURF study

Background: Urinary tract infections (UTIs) are prevalent world-wide, particularly among women. Their incidence increases with age, and treatment is increasingly challenging owing to antibiotic resistance and the lack of new agents. We investigated the susceptibility of current urinary isolates to fosfomycin and other antibiotics across Europe. Methods: This cross-sectional study collected consecutive urinary isolates from non-hospitalised women at 20 centres in Belgium, UK, Italy, Spain and Russia. Bacteria were tested by disk diffusion with relevant antibiotics. As a quality control, a central laboratory re-tested, by agar dilution: (i) isolates found resistant to fosfomycin, and (ii) every tenth isolate; all non-Russian sites were included. Results: A total of 2848 isolates were analysed, principally Escherichia coli (2064, 72.5%), Klebsiella spp. (275, 9.7%) and 103 Proteus spp. (103, 3.6%). For E. coli, agents active against >90% of isolates were nitrofurantoin (98.5%), fosfomycin (96.4%), and mecillinam (91.8%). Fosfomycin and nitrofurantoin remained active against >90% of cephalosporin-resistant E. coli. Among 143 E. coli recorded as susceptible locally by disk tests, 138 (96.5%) were confirmed susceptible by MIC tests, however resistance was only confirmed in 29/58 (50%) of those reported resistant by local disk tests. Conclusion: E. coli was found to be the most common uropathogen isolated and was highly susceptible to fosfomycin, nitrofurantoin and mecillinam, all used effectively for more than 30 years. Guidelines advocating fosfomycin for uUTIs in women remain microbiologically valid