Two Novel Fish Paralogs Provide Insights Into the Rid Family of Imine Deaminases Active in Pre-Empting enamine/imine Metabolic Damage

Reactive Intermediate Deaminase (Rid) protein superfamily includes eight families among which the RidA is conserved in all domains of life. RidA proteins accelerate the deamination of the reactive 2-aminoacrylate (2AA), an enamine produced by some pyridoxal phosphate (PLP)-dependent enzymes. 2AA accumulation inhibits target enzymes with a detrimental impact on fitness. As a consequence of whole genome duplication, teleost fish have two ridA paralogs, while other extant vertebrates contain a single-copy gene. We investigated the biochemical properties of the products of two paralogs, identified in Salmo salar. SsRidA-1 and SsRidA-2 complemented the growth defect of a Salmonella enterica ridA mutant, an in vivo model of 2AA stress. In vitro, both proteins hydrolyzed 2-imino acids (IA) to keto-acids and ammonia. SsRidA-1 was active on IA derived from nonpolar amino acids and poorly active or inactive on IA derived from other amino acids tested. In contrast, SsRidA-2 had a generally low catalytic efficiency, but showed a relatively higher activity with IA derived from L-Glu and aromatic amino acids. The crystal structures of SsRidA-1 and SsRidA-2 provided hints of the remarkably different conformational stability and substrate specificity. Overall, SsRidA-1 is similar to the mammalian orthologs whereas SsRidA-2 displays unique properties likely generated by functional specialization of a duplicated ancestral gene

Repurposing the trypanosomatidic gsk kinetobox for the inhibition of parasitic pteridine and dihydrofolate reductases

Three open-source anti-kinetoplastid chemical boxes derived from a whole-cell phenotypic screening by GlaxoSmithKline (Tres Cantos Anti-Kinetoplastid Screening, TCAKS) were exploited for the discovery of a novel core structure inspiring new treatments of parasitic diseases targeting the trypansosmatidic pteridine reductase 1 (PTR1) and dihydrofolate reductase (DHFR) enzymes. In total, 592 compounds were tested through medium-throughput screening assays. A subset of 14 compounds successfully inhibited the enzyme activity in the low micromolar range of at least one of the enzymes from both Trypanosoma brucei and Lesihmania major parasites (pan-inhibitors), or from both PTR1 and DHFR-TS of the same parasite (dual inhibitors). Molecular docking studies of the protein–ligand interaction focused on new scaffolds not reproducing the well-known antifolate core clearly explaining the experimental data. TCMDC-143249, classified as a benzenesulfonamide derivative by the QikProp descriptor tool, showed selective inhibition of PTR1 and growth inhibition of the kinetoplastid parasites in the 5 µM range. In our work, we enlarged the biological profile of the GSK Kinetobox and identified new core structures inhibiting selectively PTR1, effective against the kinetoplastid infectious protozoans. In perspective, we foresee the development of selective PTR1 and DHFR inhibitors for studies of drug combinations

The GINGER Project and status of the ring-laser of LNGS

A ring-laser attached to the Earth measures the absolute angular velocity of the Earth summed to the relativistic precessions, de Sitter and Lense-Thirring. GINGER (Gyroscopes IN GEneral Relativity) is a project aiming at measuring the LenseThirring effect with a ground based detector; it is based on an array of ring-lasers. Comparing the Earth angular velocity measured by IERS and the measurement done with the GINGER array, the Lense-Thirring effect can be evaluated. Compared to the existing space experiments, GINGER provides a local measurement, not the averaged value and it is unnecessary to model the gravitational field. It is a proposal, but it is not far from being a reality. In fact the GrossRing G of the Geodesy Observatory of Wettzell has a sensitivity very close to the necessary one. G ofWettzell is part of the IERS system which provides the measure of the Length Of the DAY (LOD); G provides information on the fast component of LOD. In the last few years, a roadmap toward GINGER has been outlined. The experiment G-GranSasso, financed by the INFN Commission II, is developing instrumentations and tests along the roadmap of GINGER. In this short paper the main activities of G-GranSasso and some results will be presented. The first results of GINGERino will be reported, GINGERino is the large ring-laser installed inside LNGS and now in the commissioning phase. Ring-lasers provide as well important informations for geophysics, in particular the rotational seismology, which is an emerging field of science. GINGERino is one of the three experiments of common interest between INFN and INGV

Measurement results and improvements on an open EPR system

Electron spin resonance (ESR) is a spectroscopic method that allows to measure stable radicals induced by ionizing radiation. The EPR measurements can help to estimate the dose absorbed by people exposed during a nuclear disaster, detecting the number of radicals induced in their mobile phones due to the exposition [1]. Using conventional closed microwave cavities, the phone display must be fragmented in order to be introduced inside the resonator, becoming no more usable. The aim of this work is to develop a system, compatible with the spectrometer Bruker Elexys E500, able to preserve the sample integrity. The system uses an X-band resonant metallic cavity with a slit, realized on one side, for the leak of the excitation magnetic field and a Helmotz coil pair. The resonator allows measuring a sample lodged outside the cavity, while the coils produce a 100 kHz modulated field that encodes the output signal at a particular frequency and increases the SNR