Label-Free Mass Spectrometry Proteomics Reveals Different Pathways Modulated in THP-1 Cells Infected with Therapeutic Failure and Drug Resistance Leishmania infantum Clinical Isolates

As the world is facing increasing difficulties to treat leishmaniasis with current therapies, deeper investigation into the molecular mechanisms responsible for both drug resistance and treatment failure (TF) is essential in drug discovery and development. So far, few available drugs cause severe side effects and have developed several resistance mechanisms. Drug resistance and TF parasite strains from clinical isolates may have acquired altered expression of proteins that characterize specific mechanisms leading to therapy inefficacy. This work aims to identify the biochemical pathways of THP-1 human monocytes infected by different Leishmania infantum clinical isolates from patients with either resistance or with TF outcome, using whole cell differential Mass Spectrometry proteomics. We have adopted network enrichment analysis to integrate the transcriptomics and the proteomic results of infected cells studies. Transferrin receptor C (TFRC) and nucleoside diphosphate kinase 3 (NDK3) were discovered as overexpressed proteins in THP-1 cells infected with paromomycin, antimony, and miltefosine resistant L. infantum lines. The overall achievements represent founding concepts to confirm new targets involved in the parasitic drug resistance and TF mechanisms, and to consider in perspective the importance of a dual host-guest pharmacological approach to treat the acute stage of the disease.We thank Dr. F. Javier Moreno from the WHO Collaborating Center for Leishmaniasis, Instituto de Salud Carlos III (ISCIII), for providing L. infantum lines LLM2070, LLM2165, LLM2255, and LLM2221, isolated from HIV-positive patients with visceral leishmaniasis and TF, and the paromomycin-resistant L. infantum line LEM2126 (L2126) used in this study. Also, we thank Dra. Laurence Lachaud from the Centre National de Référence des Leishmanioses, Université Montpellier (Montpellier, France), for providing the drug-resistant L. infantum lines used in this work: LEM3323 (L3323) and LEM5159 (L5159), which are SbIII- and Mil-resistant lines, respectively. The authors thank Dr. Stefania Ferrari for compiling the Leishmania database for protein search. L. infantum db was obtained from SwissProt (L. infantum entry, exported in FASTA format, updated January 2016, updates are ongoing). This work was supported in part by Grant RTI2018-097210-B-100 (to F.G.), funded by MCIN/AEI/10.13039/501100011033 and by “ERDF A Way of Making Europe” and by Grant FP7-HEALTH-2013-INNOVATION “New Medicine for Trypanosomatidic Infections” (Grant 603240). The authors acknowledge the “Fondazione Cassa di Risparmio di Modena” for funding the UHPLC-ESI-HRMS Q-Exactive system at the Centro Interdipartimentale Grandi Strumenti (CIGS) of the University of Modena and Reggio Emilia. The authors thank the COST Action “OneHealthdrugs” CA21111 for inspiring the research development

Search for Permanent Electric Dipole Moments at COSY Step 1: Spin coherence and systematic error studies

Permanent EDMs (Electric Dipole Moments) of fundamental particle violate both time invariance and parity. Assuming the CPT theorem this implies CP violation. The standard model predicts non-vanishing EDMs, their magnitudes, however, are expected to be unobservably small. Hence, the discovery of a non-zero EDM would be a signal for “new physics”. EDM experiments with charged particles are only possible at storage rings. As a first step towards EDM searches in storage rings, pursued by the recently founded JEDI collaboration, we propose research and development work to be carried out at COSY to maximize the spin coherence time and to reduce systematic spin rotations. Both aspects focus on a first direct measurement of a charged particle EDM in a storage ring at COSY aiming at an upper limit of ≈ 1024 e·cm, and on a longer time scale to reach an even higher sensitivity constructing a dedicated storage ring

Tocilizumab for patients with COVID-19 pneumonia. The single-arm TOCIVID-19 prospective trial

BackgroundTocilizumab blocks pro-inflammatory activity of interleukin-6 (IL-6), involved in pathogenesis of pneumonia the most frequent cause of death in COVID-19 patients.MethodsA multicenter, single-arm, hypothesis-driven trial was planned, according to a phase 2 design, to study the effect of tocilizumab on lethality rates at 14 and 30 days (co-primary endpoints, a priori expected rates being 20 and 35%, respectively). A further prospective cohort of patients, consecutively enrolled after the first cohort was accomplished, was used as a secondary validation dataset. The two cohorts were evaluated jointly in an exploratory multivariable logistic regression model to assess prognostic variables on survival.ResultsIn the primary intention-to-treat (ITT) phase 2 population, 180/301 (59.8%) subjects received tocilizumab, and 67 deaths were observed overall. Lethality rates were equal to 18.4% (97.5% CI: 13.6-24.0, P=0.52) and 22.4% (97.5% CI: 17.2-28.3, P<0.001) at 14 and 30 days, respectively. Lethality rates were lower in the validation dataset, that included 920 patients. No signal of specific drug toxicity was reported. In the exploratory multivariable logistic regression analysis, older age and lower PaO2/FiO2 ratio negatively affected survival, while the concurrent use of steroids was associated with greater survival. A statistically significant interaction was found between tocilizumab and respiratory support, suggesting that tocilizumab might be more effective in patients not requiring mechanical respiratory support at baseline.ConclusionsTocilizumab reduced lethality rate at 30 days compared with null hypothesis, without significant toxicity. Possibly, this effect could be limited to patients not requiring mechanical respiratory support at baseline.Registration EudraCT (2020-001110-38); clinicaltrials.gov (NCT04317092)