Synthesis and screening of 6‐alkoxy purine analogs as cell type‐selective apoptotic inducers in Jurkat cells

Ministerio de Ciencia e Innovacion, Grant/Award Number: RTC-2017-6620; Ministerio de Educacion, Cultura y Deporte, Grant/Award Number: FPU 14/00818Purines are ubiquitous structures in cell biology involved in a multitude of cellular processes, because of which substituted purines and analogs are considered excellent scaffolds in drug design. In this study, we explored the key structural features of a purine‐based proapoptotic hit, 8‐tert‐butyl‐9‐phenyl‐6‐benzyloxy‐9Hpurine (1), by setting up a library of 6‐alkoxy purines with the aim of elucidating the structural requirements that govern its biological activity and to study the cell selectivity of this chemotype. This was done by a phenotypic screening approach based on cell cycle analysis of a panel of six human cancer cell lines, including T cell leukemia Jurkat cells. From this study, two derivatives (12 and 13) were identified as Jurkat‐selective proapoptotic compounds, displaying superior potency and cell selectivity than hit 1.Instituto de Salud Carlos III Spanish Government European Commission RTC-2017-6620Ministerio de Educacion, Cultura y Deporte FPU 14/0081

Randomised multicentre clinical trial to evaluate voriconazole pre-emptive genotyping strategy in patients with risk of aspergillosis: vorigenipharm study protocol.

Introduction Invasive aspergillosis is the most important cause of morbidity and mortality in patients with haematological diseases. At present, voriconazole is the first-line treatment for invasive fungal disease. The pharmacokinetic interindividual variability of voriconazole depends on genetic factors. CYP450 is involved in 70%–75% of total metabolism of voriconazole, mainly CYP3A4 and CYP2C19, with the remaining 25%–30% of metabolism conducted by monooxygenase flavins. CYP2C19 single nucleotide polymorphisms could explain 50%–55% of variability in voriconazole metabolism. Materials and methods The main objective is to compare efficiency of pre-emptive voriconazole genotyping with routine practice. The primary outcome is serum voriconazole on the fifth day within the therapeutic range. The secondary outcome is the combined variables of therapeutic failure and adverse events within 90 days of first administration, associated with voriconazole. A total of 146 patients at risk of invasive aspergillosis who will potentially receive voriconazole will be recruited, and CYP2C19 will be genotyped. If the patient ultimately receives voriconazole, they will be randomised (1:1 experimental/control). In the experimental arm, patients will receive a dose according to a pharmacogenetic algorithm, including CYP2C19 genotype and clinical and demographic information. In the control arm, patients will receive a dose according to clinical practice guidelines. In addition, a Spanish National Healthcare System (NHS) point-of-view cost-effectiveness evaluation will be performed. Direct cost calculations for each arm will be performed. Conclusion This trial will provide information about the viability and cost-effectiveness of the mplementation of a pre-emptive voriconazole genotyping strategy in the Spanish NHS. Ethics and dissemination A Spanish version of this protocol has been evaluated and approved by the La Paz University Hospital Ethics Committee and the Spanish Agency of Medicines and Medical Devices. Trial results will be submitted for publication in an open peer-reviewed medical speciality-specific publication. Trial registration number Eudra-CT: 2019-000376-41 and NCT04238884; Pre-results.post-print441 K

RNA-binding proteins regulate cell respiration and coenzyme Q biosynthesis by post-transcriptional regulation of COQ7

et al.Coenzyme Q (CoQ) is a key component of the mitochondrial respiratory chain carrying electrons from complexes I and II to complex III and it is an intrinsic component of the respirasome. CoQ concentration is highly regulated in cells in order to adapt the metabolism of the cell to challenges of nutrient availability and stress stimuli. At least 10 proteins have been shown to be required for CoQ biosynthesis in a multi-peptide complex and COQ7 is a central regulatory factor of this pathway. We found that the first 765 bp of the 3′-untranslated region (UTR) of COQ7 mRNA contains cis-acting elements of interaction with RNA-binding proteins (RBPs) HuR and hnRNP C1/C2. Binding of hnRNP C1/C2 to COQ7 mRNA was found to require the presence of HuR, and hnRNP C1/C2 silencing appeared to stabilize COQ7 mRNA modestly. By contrast, lowering HuR levels by silencing or depriving cells of serum destabilized and reduced the half-life of COQ7 mRNA, thereby reducing COQ7 protein and CoQ biosynthesis rate. Accordingly, HuR knockdown decreased oxygen consumption rate and mitochondrial production of ATP, and increased lactate levels. Taken together, our results indicate that a reduction in COQ7 mRNA levels by HuR depletion causes mitochondrial dysfunction and a switch toward an enhanced aerobic glycolysis, the characteristic phenotype exhibited by primary deficiency of CoQ. Thus HuR contributes to efficient oxidative phosphorylation by regulating of CoQ biosynthesis.This research was supported by grants from the Spanish Ministry of Health, Instituto de Salud Carlos III (ISCIII), FIS PI14–01962 to PN, the Spanish Ministry of Economy and Competitively, SAF2013–41945-R to JMC, and the Intramural Research Program of the NIA, NIH (KA, MG).Peer Reviewe

Decreased oxytocin plasma levels and oxytocin receptor expression in borderline personality disorder

Introduction: Borderline personality disorder (BPD) is characterized by intense affective reactions with underlying social and interpersonal cognitive deficits. Oxytocin has largely been associated with both stress regulation and social cognition in psychiatric patients and in non-clinical populations in previous studies. Finally, abnormal oxytocin levels have been preliminary reported in BPD patients. Methods: 53 patients with moderate-severe BPD and 31 healthy control subjects were investigated for plasma levels of oxytocin and protein expression of oxytocin receptor in blood mononuclear cells. Clinical assessments were made for severity, functionality, and comorbidity with axis I and II conditions. Results: Oxytocin plasma levels were significantly lower in BPD patients compared with controls. In addition, protein expression of oxytocin receptor was significantly reduced in the BPD group. A positive correlation was found between plasma oxytocin levels and the activity index score of the Zuckerman-Kuhlman Personality Questionnaire (ZKPQ). Oxytocin receptor protein expression, on the contrary, had a negative correlation with the ZKPQ sociability index score. Conclusions: Results support the evidence of a dysfunction of the oxytocin system in borderline personality disorder, which could be involved in emotional dysregulation and interpersonal disturbances in these patients.Instituto de Salud Carlos IIIFEDERDepto. de Farmacología y ToxicologíaDepto. de Medicina Legal, Psiquiatría y PatologíaFac. de MedicinaTRUEpu

Transdiagnostic study of impulsivity and self-injurious behaviour in unstable and impulsive disorders

Introduction. High comorbidity between borderline personality disorder (BPD) and eating disorders (ED) shows the necessity of developing transdiagnostic models, where impulsivity could play a relevant role in the manifestations of self-injurious behaviour. Objectives: 1) To compare the levels of impulsivity and self-injurious behaviour among people with BPD, people with ED and controls. 2) To predict the presence of self-injurious behaviour based on impulsivity and other relevant clinical variables, such as the experience of traumatic events and sensitivity to rejection. Methods. 108 women (23 controls; 29 with a diagnosis of restrictive ACT (rED); 21 with a diagnosis of purgative ACT (pED); and 35 with a diagnosis of BPD) were assessed using the Barratt Impulsivity Scale, the Traumatic Experiences Questionnaire and the Sensitivity to Rejection Questionnaire. Information about self-injurious behaviour was collected through interview and clinical history. Results. Differences in impulsivity and self-injurious behaviour scores were found between the groups, with higher impulsivity in the BPD and pED groups, and higher rates of self-injurious behaviour in the BPD group followed by both ED groups. On the other hand, impulsivity predicted suicide attempts, and together with sensitivity to interpersonal rejection predicted nonsuicidal self-harm (NSSH). Conclusion. Impulsivity is a dimensional variable in BPD and ED, which in turn plays a relevant role in the prediction of self-injurious behaviour.Acknowledgements and conflicts of interest. This work has been supported by the project PI16/01949, integrated in the Plan Nacional de I+D+I, AES 2013-2016; funded by the ISCIII and co-financed by the Fondo Europeo de Desarrollo Regional (FEDER). “Una forma de hacer Europa

Dielectron and heavy-quark production in inelastic and high-multiplicity proton–proton collisions at √s = 13 TeV

The measurement of dielectron production is presented as a function of invariant mass and transverse momentum (pT) at midrapidity (|ye| < 0.8) in proton–proton (pp) collisions at a centre-of-mass energy of √s = 13 TeV. The contributions from light-hadron decays are calculated from their measured cross sections in pp collisions at √s = 7 TeV or 13 TeV. The remaining continuum stems from correlated semileptonic decays of heavy-flavour hadrons. Fitting the data with templates from two different MC event generators, PYTHIA and POWHEG, the charm and beauty cross sections at midrapidity are extracted for the first time at this collision energy: dσcc¯/dy|y=0 = 974 ± 138 (stat.) ± 140 (syst.) ± 214(BR) μb and dσbb¯ /dy|y=0 = 79 ± 14 (stat.) ± 11 (syst.) ± 5(BR) μb using PYTHIA simulations and dσcc¯/dy|y=0 = 1417 ± 184 (stat.) ± 204 (syst.) ± 312(BR) μb and dσbb¯ /dy|y=0 = 48 ± 14 (stat.) ± 7 (syst.) ± 3(BR) μb for POWHEG. These values, whose uncertainties are fully correlated between the two generators, are consistent with extrapolations from lower energies. The different results obtained with POWHEG and PYTHIA imply different kinematic correlations of the heavy-quark pairs in these two generators. Furthermore, comparisons of dielectron spectra in inelastic events and in events collected with a trigger on high charged-particle multiplicities are presented in various pT intervals. The differences are consistent with the already measured scaling of light-hadron and open-charm production at high charged-particle multiplicity as a function of pT. Upper limits for the contribution of virtual direct photons are extracted at 90% confidence level and found to be in agreement with pQCD calculations

Charged-particle multiplicity distributions over a wide pseudorapidity range in proton-proton collisions at √s = 0.9, 7 and 8 TeV

We present the charged-particle multiplicity distributions over a wide pseudorapidity range (−3.4<η<5.0) for pp collisions at s√= 0.9, 7, and 8 TeV at the LHC. Results are based on information from the Silicon Pixel Detector and the Forward Multiplicity Detector of ALICE, extending the pseudorapidity coverage of the earlier publications and the high-multiplicity reach. The measurements are compared to results from the CMS experiment and to PYTHIA, PHOJET and EPOS LHC event generators, as well as IP-Glasma calculations

Constraining the magnitude of the Chiral Magnetic Effect with Event Shape Engineering in Pb-Pb collisions at sNN\sqrt{s_{\rm NN}} = 2.76$ TeV

In ultrarelativistic heavy-ion collisions, the event-by-event variation of the elliptic flow $v_2$ reflects fluctuations in the shape of the initial state of the system. This allows to select events with the same centrality but different initial geometry. This selection technique, Event Shape Engineering, has been used in the analysis of charge-dependent two- and three-particle correlations in Pb-Pb collisions at $\sqrt{s_{_{\rm NN}}} =2.76$ TeV. The two-particle correlator $\langle \cos(\varphi_\alpha - \varphi_\beta) \rangle$, calculated for different combinations of charges $\alpha$ and $\beta$, is almost independent of $v_2$ (for a given centrality), while the three-particle correlator $\langle \cos(\varphi_\alpha + \varphi_\beta - 2\Psi_2) \rangle$ scales almost linearly both with the event $v_2$ and charged-particle pseudorapidity density. The charge dependence of the three-particle correlator is often interpreted as evidence for the Chiral Magnetic Effect (CME), a parity violating effect of the strong interaction. However, its measured dependence on $v_2$ points to a large non-CME contribution to the correlator. Comparing the results with Monte Carlo calculations including a magnetic field due to the spectators, the upper limit of the CME signal contribution to the three-particle correlator in the 10-50% centrality interval is found to be 26-33% at 95% confidence level

Constraining the magnitude of the chiral magnetic effect with event shape engineering in Pb–Pb collisions at √sNN=2.76 TeV

In ultrarelativistic heavy-ion collisions, the event-by-event variation of the elliptic flow v2 reflects fluctuations in the shape of the initial state of the system. This allows to select events with the same centrality but different initial geometry. This selection technique, Event Shape Engineering, has been used in the analysis of charge-dependent two- and three-particle correlations in Pb–Pb collisions at √sNN=2.76 TeV. The two-particle correlator 〈cos⁡(φα−φβ)〉, calculated for different combinations of charges α and β, is almost independent of v2 (for a given centrality), while the three-particle correlator 〈cos⁡(φα+φβ−2Ψ2)〉 scales almost linearly both with the event v2 and charged-particle pseudorapidity density. The charge dependence of the three-particle correlator is often interpreted as evidence for the Chiral Magnetic Effect (CME), a parity violating effect of the strong interaction. However, its measured dependence on v2 points to a large non-CME contribution to the correlator. Comparing the results with Monte Carlo calculations including a magnetic field due to the spectators, the upper limit of the CME signal contribution to the three-particle correlator in the 10–50% centrality interval is found to be 26–33% at 95% confidence level