The assessment of the potential hepatotoxicity of new drugs by in vitro metabolomics

Drug hepatotoxicity assessment is a relevant issue both in the course of drug development as well as in the post marketing phase. The use of human relevant in vitro models in combination with powerful analytical methods (metabolomic analysis) is a promising approach to anticipate, as well as to understand and investigate the effects and mechanisms of drug hepatotoxicity in man. The metabolic profile analysis of biological liver models treated with hepatotoxins, as compared to that of those treated with non-hepatotoxic compounds, provides useful information for identifying disturbed cellular metabolic reactions, pathways, and networks. This can later be used to anticipate, as well to assess, the potential hepatotoxicity of new compounds. However, the applicability of the metabolomic analysis to assess the hepatotoxicity of drugs is complex and requires careful and systematic work, precise controls, wise data preprocessing and appropriate biological interpretation to make meaningful interpretations and/or predictions of drug hepatotoxicity. This review provides an updated look at recent in vitro studies which used principally mass spectrometry-based metabolomics to evaluate the hepatotoxicity of drugs. It also analyzes the principal drawbacks that still limit its general applicability in safety assessment screenings. We discuss the analytical workflow, essential factors that need to be considered and suggestions to overcome these drawbacks, as well as recent advancements made in this rapidly growing field of research

Low-lying magnetic excitations of doubly-closed-shell nuclei and nucleon-nucleon effective interactions

We have studied the low lying magnetic spectra of 12C, 16O, 40Ca, 48Ca and 208Pb nuclei within the Random Phase Approximation (RPA) theory, finding that the description of low-lying magnetic states of doubly-closed-shell nuclei imposes severe constraints on the spin and tensor terms of the nucleon-nucleon effective interaction. We have first made an investigation by using four phenomenological effective interactions and we have obtained good agreement with the experimental magnetic spectra, and, to a lesser extent, with the electron scattering responses. Then we have made self-consistent RPA calculations to test the validity of the finite-range D1 Gogny interaction. For all the nuclei under study we have found that this interaction inverts the energies of all the magnetic states forming isospin doublets.Comment: 19 pages, 13 figures, 7 tables, accepted for publication in Phys. Rev.

Factors that influence the quality of metabolomics data in in vitro cell toxicity studies: a systematic survey

REACH (Registration, Evaluation, Authorization and Restriction of Chemicals) is a global strategy and regulation policy of the EU that aims to improve the protection of human health and the environment through the better and earlier identification of the intrinsic properties of chemical substances. It entered into force on 1st June 2007 (EC 1907/2006). REACH and EU policies plead for the use of robust high-throughput 'omic' techniques for the in vitro investigation of the toxicity of chemicals that can provide an estimation of their hazards as well as information regarding the underlying mechanisms of toxicity. In agreement with the 3R's principles, cultured cells are nowadays widely used for this purpose, where metabolomics can provide a real-time picture of the metabolic effects caused by exposure of cells to xenobiotics, enabling the estimations about their toxicological hazards. High quality and robust metabolomics data sets are essential for precise and accurate hazard predictions. Currently, the acquisition of consistent and representative metabolomic data is hampered by experimental drawbacks that hinder reproducibility and difficult robust hazard interpretation. Using the differentiated human liver HepG2 cells as model system, and incubating with hepatotoxic (acetaminophen and valproic acid) and non-hepatotoxic compounds (citric acid), we evaluated in-depth the impact of several key experimental factors (namely, cell passage, processing day and storage time, and compound treatment) and instrumental factors (batch effect) on the outcome of an UPLC-MS metabolomic analysis data set. Results showed that processing day and storage time had a significant impact on the retrieved cell's metabolome, while the effect of cell passage was minor. Meta-analysis of results from pathway analysis showed that batch effect corrections and quality control (QC) measures are critical to enable consistent and meaningful estimations of the effects caused by compounds on cells. The quantitative analysis of the changes in metabolic pathways upon bioactive compound treatment remained consistent despite the concurrent causes of metabolomic data variation. Thus, upon appropriate data retrieval and correction and by an innovative metabolic pathway analysis, the metabolic alteration predictions remained conclusive despite the acknowledged sources of variability

Balancing porosity and mechanical properties of titanium samples to favor cellular growth against bacteria

Two main problems limit the success of titanium implants: bacterial infection, which restricts their osseointegration capacity; and the stiffness mismatch between the implant and the host cortical bone, which promotes bone resorption and risk of fracture. Porosity incorporation may reduce this difference in stiffness but compromise biomechanical behavior. In this work, the relationship between the microstructure (content, size, and shape of pores) and the antibacterial and cellular behavior of samples fabricated by the space-holder technique (50 vol % NH4HCO3 and three ranges of particle sizes) is established. Results are discussed in terms of the best biomechanical properties and biofunctional activity balance (cell biocompatibility and antibacterial behavior). All substrates achieved suitable cell biocompatibility of premioblast and osteoblast in adhesion and proliferation processes. It is worth to highlighting that samples fabricated with the 100–200 μm space-holder present better mechanical behavior—in terms of stiffness, microhardness, and yield strength—which make them a very suitable material to replace cortical bone tissues. Those results exposed the relationship between the surface properties and the race of bacteria and mammalian cells for the surface with the aim to promote cellular growth over bacteria.University of Seville (Spain) VI Plan Propio de Investigación y Transferencia—US 2018, I.3A

Comparing Targeted vs. Untargeted MS2 Data-Dependent Acquisition for Peak Annotation in LC-MS Metabolomics

One of the most widely used strategies for metabolite annotation in untargeted LCMS is based on the analysis of MSn spectra acquired using data-dependent acquisition (DDA), where precursor ions are sequentially selected from MS scans based on user-selected criteria. However, the number of MSn spectra that can be acquired during a chromatogram is limited and a trade-off between analytical speed, sensitivity and coverage must be ensured. In this research, we compare four different strategies for automated MS2 DDA, which can be easily implemented in the frame of standard QA/QC workflows for untargeted LC-MS. These strategies consist of (i) DDA in the MS working range; (ii) iterated DDA split into several m/z intervals; (iii) dynamic iterated DDA of (pre)selected potentially informative features; and (iv) dynamic iterated DDA of (pre)annotated metabolic features using a reference database. Their performance was assessed using the analysis of human milk samples as model example by comparing the percentage of LC-MS features selected as the precursor ion for MS2, the number, and class of annotated features, the speed and confidence of feature annotation, and the number of LC runs required

A beam-beam monitoring detector for the MPD experiment at NICA

The Multi-Purpose Detector (MPD) is to be installed at the Nuclotron Ion Collider fAcility (NICA) of the Joint Institute for Nuclear Research (JINR). Its main goal is to study the phase diagram of the strongly interacting matter produced in heavy-ion collisions. These studies, while providing insight into the physics of heavy-ion collisions, are relevant for improving our understanding of the evolution of the early Universe and the formation of neutron stars. In order to extend the MPD trigger capabilities, we propose to include a high granularity beam-beam monitoring detector (BE-BE) to provide a level-0 trigger signal with an expected time resolution of 30 ps. This new detector will improve the determination of the reaction plane by the MPD experiment, a key measurement for flow studies that provides physics insight into the early stages of the reaction. In this work, we use simulated Au+Au collisions at NICA energies to show the potential of such a detector to determine the event plane resolution, providing further redundancy to the detectors originally considered for this purpose namely, the Fast Forward Detector (FFD) and the Hadron Calorimeter (HCAL). We also show our results for the time resolution studies of two prototype cells carried out at the T10 beam line at the CERN PS complex.Comment: 16 pages, 12 figures. Updated to published version with added comments and correction

MAGIC Observations of the Nearby Short Gamma-Ray Burst GRB 160821B

Acciari, A. V., et al.The coincident detection of GW170817 in gravitational waves and electromagnetic radiation spanning the radio to MeV gamma-ray bands provided the first direct evidence that short gamma-ray bursts (GRBs) can originate from binary neutron star (BNS) mergers. On the other hand, the properties of short GRBs in high-energy gamma-rays are still poorly constrained, with only ∼20 events detected in the GeV band, and none in the TeV band. GRB 160821B is one of the nearest short GRBs known at z = 0.162. Recent analyses of the multiwavelength observational data of its afterglow emission revealed an optical-infrared kilonova component, characteristic of heavy-element nucleosynthesis in a BNS merger. Aiming to better clarify the nature of short GRBs, this burst was automatically followed up with the MAGIC telescopes, starting from 24 s after the burst trigger. Evidence of a gamma-ray signal is found above ∼0.5 TeV at a significance of ∼ 3σ during observations that lasted until 4 hr after the burst. Assuming that the observed excess events correspond to gamma-ray emission from GRB 160821B, in conjunction with data at other wavelengths, we investigate its origin in the framework of GRB afterglow models. The simplest interpretation with one-zone models of synchrotron-self-Compton emission from the external forward shock has difficulty accounting for the putative TeV flux. Alternative scenarios are discussed where the TeV emission can be relatively enhanced. The role of future GeV-TeV observations of short GRBs in advancing our understanding of BNS mergers and related topics is briefly addressed.We would like to thank the Instituto de Astrofísica de Canarias for the excellent working conditions at the Observatorio del Roque de los Muchachos in La Palma. The financial support of the German BMBF and MPG; the Italian INFN and INAF; the Swiss National Fund SNF; the ERDF under the Spanish MINECO (FPA2017-87859-P, FPA2017-85668-P, FPA2017- 82729-C6-2-R, FPA2017-82729-C6-6-R, FPA2017-82729-C6-5- R, AYA2015-71042-P, AYA2016-76012-C3-1-P, ESP2017- 87055-C2-2-P, FPA2017-90566-REDC); the Indian Department of Atomic Energy; the Japanese ICRR, the University of Tokyo, JSPS, and MEXT; the Bulgarian Ministry of Education and Science, National RI Roadmap Project DO1-268/16.12.2019 and the Academy of Finland grant No. 320045 is gratefully acknowledged. This work was also supported by the Spanish Centro de Excelencia “Severo Ochoa” SEV-2016-0588, SEV2015-0548 and SEV-2012-0234, the Unidad de Excelencia “María de Maeztu” MDM-2014-0369 and the “la Caixa” Foundation (fellowship LCF/BQ/PI18/11630012), by the Croatian Science Foundation (HrZZ) Project IP-2016-06-9782 and the University of Rijeka Project 13.12.1.3.02, by the DFG Collaborative Research Centers SFB823/C4 and SFB876/C3, the Polish National Research Centre grant UMO-2016/22/M/ST9/ 00382 and by the Brazilian MCTIC, CNPq and FAPERJ. K.N. is thankful for the support by Marie Skłodowska-Curie actions (H2020-MSCA-COFUND-2014, Project P-Sphere GA 665919), and JSPS KAKENHI grant No. JP20KK0067 from MEXT, Japan. L.N. acknowledges funding from the European Union’s Horizon 2020 research and innovation program under the Marie Skłodowska-Curie grant agreement No. 664931. S.I. is supported by JSPS KAKENHI grant No. JP17K05460 from MEXT, Japan, and the RIKEN iTHEMS program

Search for Very High-energy Emission from the Millisecond Pulsar PSR J0218+4232

Acciari, V. A., et al. (MAGIC Collaboration)PSR J0218+4232 is one of the most energetic millisecond pulsars known and has long been considered as one of the best candidates for very high-energy (VHE; >100 GeV) ?-ray emission. Using 11.5 yr of Fermi Large Area Telescope (LAT) data between 100 MeV and 870 GeV, and ~90 hr of Major Atmospheric Gamma Imaging Cherenkov (MAGIC) observations in the 20 GeV to 20 TeV range, we searched for the highest energy ?-ray emission from PSR J0218+4232. Based on the analysis of the LAT data, we find evidence for pulsed emission above 25 GeV, but see no evidence for emission above 100 GeV (VHE) with MAGIC. We present the results of searches for ?-ray emission, along with theoretical modeling, to interpret the lack of VHE emission. We conclude that, based on the experimental observations and theoretical modeling, it will remain extremely challenging to detect VHE emission from PSR J0218+4232 with the current generation of Imaging Atmospheric Cherenkov Telescopes, and maybe even with future ones, such as the Cherenkov Telescope Array.The financial support of the German BMBF, MPG and HGF; the Italian INFN and INAF; the Swiss National Fund SNF; the ERDF under the Spanish Ministerio de Ciencia e Innovación (MICINN) (FPA2017-87859-P, FPA2017-85668-P, FPA2017-82729-C6-5-R, FPA2017-90566- REDC, PID2019-104114RB-C31, PID2019-104114RB-C32, PID2019-105510GB-C31,PID2019-107847RB-C41, PID2019- 107847RB-C42, PID2019-107988GB-C22); the Indian Department of Atomic Energy; the Japanese ICRR, the University of Tokyo, JSPS, and MEXT; the Bulgarian Ministry of Education and Science, National RI Roadmap Project DO1-268/16.12.2019 and the Academy of Finland grant No. 320045 is gratefully acknowledged. This work was also supported by the Spanish Centro de Excelencia “Severo Ochoa” SEV-2016-0588 and CEX2019-000920-S, and “María de Maeztu” CEX2019-000918- M, the Unidad de Excelencia “María de Maeztu” MDM-2015- 0509-18-2 and the “la Caixa” Foundation (fellowship LCF/BQ/ PI18/11630012) and by the CERCA program of the Generalitat de Catalunya; by the Croatian Science Foundation (HrZZ) Project IP2016-06-9782 and the University of Rijeka Project 13.12.1.3.02; by the DFG Collaborative Research Centers SFB823/C4 and SFB876/C3; the Polish National Research Centre grant UMO2016/22/M/ST9/00382; and by the Brazilian MCTIC, CNPq and FAPERJ