Novel Regioselective Synthesis of 1,3,4,5-Tetrasubstituted Pyrazoles and Biochemical Valuation on F1FO-ATPase and Mitochondrial Permeability Transition Pore Formation

An efficient, eco-compatible, and very cheap method for the construction of fully substituted pyrazoles (Pzs) via eliminative nitrilimine-alkene 1,3-dipolar cycloaddition (ENAC) reaction was developed in excellent yield and high regioselectivity. Enaminones and nitrilimines generated in situ were selected as dipolarophiles and dipoles, respectively. A deep screening of the employed base, solvent, and temperature was carried out to optimize reaction conditions. Recycling tests of ionic liquid were performed, furnishing efficient performance until six cycles. Finally, a plausible mechanism of cycloaddition was proposed. Then, the effect of three different structures of Pzs was evaluated on the F1FO-ATPase activity and mitochondrial permeability transition pore (mPTP) opening. The Pz derivatives’ titration curves of 6a, 6h, and 6o on the F1FO-ATPase showed a reduced activity of 86%, 35%, and 31%, respectively. Enzyme inhibition analysis depicted an uncompetitive mechanism with the typical formation of the tertiary complex enzyme-substrate-inhibitor (ESI). The dissociation constant of the ESI complex (Ki’) in the presence of the 6a had a lower order of magnitude than other Pzs. The pyrazole core might set the specific mechanism of inhibition with the F1FO-ATPase, whereas specific functional groups of Pzs might modulate the binding affinity. The mPTP opening decreased in Pz-treated mitochondria and the Pzs’ inhibitory effect on the mPTP was concentration-dependent with 6a and 6o. Indeed, the mPTP was more efficiently blocked with 0.1 mM 6a than with 1 mM 6a. On the contrary, 1 mM 6o had stronger desensitization of mPTP formation than 0.1 mM 6o. The F1FO-ATPase is a target of Pzs blocking mPTP formation

Process design for the manufacturing of soft X-ray gratings in single-crystal diamond by high-energy heavy-ion irradiation

Artículo con 9 figurasThis paper describes in detail a novel manufacturing process for optical gratings suitable for use in the UV and soft X-ray regimes in a single-crystal diamond substrate based on highly focused swift heavy-ion irradiation. This type of grating is extensively used in light source facilities such as synchrotrons or free electron lasers, with ever-increasing demands in terms of thermal loads, depending on beamline operational parameters and architecture. The process proposed in this paper may be a future alternative to current manufacturing techniques, providing the advantage of being applicable to single-crystal diamond substrates, with their unique properties in terms of heat conductivity and radiation hardness. The paper summarizes the physical principle used for the grating patterns produced by swift heavy-ion irradiation and provides full details for the manufacturing process for a specific grating configuration, inspired in one of the beamlines at the ALBA synchrotron light source, while stressing the most challenging points for a potential implementation. Preliminary proof-of-concept experimental results are presented, showing the practical implementation of the methodology proposed herein.The authors acknowledge funding support by the following projects: PID2020-112770RB-C22 from the Spanish Ministry of Science and Innovation, TechnoFusión (III)-CM (S2018/EMT-4437) from Comunidad de Madrid (cofinanced by ERDF and ESF), agreement between Community of Madrid and Universidad Autónoma de Madrid (item “Excellence of University Professorate”). M.L.C. acknowledges financial support from the research project “Captacion de Talento UAM” Ref: #541D300 supervised by the Vice-Chancellor of Research of Universidad Autónoma de Madrid (UAM). LOREA beamline at ALBA is a project co-funded by the European Regional Development Fund (ERDF) within the Framework of the Smart Growth Operative Programme 2014-2020. The authors acknowledge the support from The Centro de Microanálisis de Materiales (CMAM)—Universidad Autónoma de Madrid, for the beam time proposal (demonstration of a grating profile for soft X-rays in diamond via ion lithography) with code IuB-005/21, and its technical staff for their contribution to the operation of the accelerator. We also acknowledge P. Olivero for very useful comments on the manuscript draf

Process design for the manufacturing of soft X-ray gratings in single-crystal diamond by high-energy heavy-ion irradiation

This paper describes in detail a novel manufacturing process for optical gratings suitable for use in the UV and soft X-ray regimes in a single-crystal diamond substrate based on highly focused swift heavy-ion irradiation. This type of grating is extensively used in light source facilities such as synchrotrons or free electron lasers, with ever-increasing demands in terms of thermal loads, depending on beamline operational parameters and architecture. The process proposed in this paper may be a future alternative to current manufacturing techniques, providing the advantage of being applicable to single-crystal diamond substrates, with their unique properties in terms of heat conductivity and radiation hardness. The paper summarizes the physical principle used for the grating patterns produced by swift heavy-ion irradiation and provides full details for the manufacturing process for a specific grating configuration, inspired in one of the beamlines at the ALBA synchrotron light source, while stressing the most challenging points for a potential implementation. Preliminary proof-of-concept experimental results are presented, showing the practical implementation of the methodology proposed herein

The Joint Action of Sesquiterpene Lactones from Leaves as an Explanation for the Activity of Cynara cardunculus

The work described herein is a continuation of a previous study centered on the bioprospect of cardoon (Cynara cardunculus) leaf extracts through the isolation of secondary metabolites with phytotoxic activity. Chromatographic fractionations of the ethyl acetate extract and spectroscopic analysis showed that the majority of the components were sesquiterpene lactones. Of these compounds, aguerin B, grosheimin, and cynaropicrin were very active on etiolated wheat coleoptile, standard target species, and weed growth. The joint action of binary mixtures of these three active sesquiterpene lactones and one nonactive compound (11,13-dihydroxy-8-desoxygrosheimin) was studied. The activities of fixed-ratio mixtures were assessed on wheat coleoptile. The results can be interpreted with respect to a reference model by considering dose−response analyses and isobolograms with linear regression analyses. A total of 17 binary mixtures at different levels of inhibition (ED25, ED50, and ED75) were studied, and predominantly they responded additively (25). Deviations from additivity included seven synergistic responses and two antagonistic responses. The joint action of major sesquiterpene lactones isolated from C. cardunculus can explain the activities observed in extracts and fractions. The results reported here reiterate the utility of the wheat coleoptile bioassay as a quick tool to detect potential synergistic effects in binary mixtures

Oral Ethanol Self-Administration in Rhesus Monkeys: Behavioral and Neurochemical Correlates

Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/66306/1/j.1530-0277.1999.tb04357.x.pd

Perifosine as a Potential Novel Anti-Cancer Agent Inhibits EGFR/MET-AKT Axis in Malignant Pleural Mesothelioma

PI3K/AKT signalling pathway is aberrantly active and plays a critical role for cell cycle progression of human malignant pleural mesothelioma (MMe) cells. AKT is one of the important cellular targets of perifosine, a novel bio-available alkylphospholipid that has displayed significant anti-proliferative activity in vitro and in vivo in several human tumour model systems and is currently being tested in clinical trials.We tested Perifosine activity on human mesothelial cells and different mesothelioma cell lines, in order to provide evidence of its efficacy as single agent and combined therapy.We demonstrate here that perifosine, currently being evaluated as an anti-cancer agent in phase 1 and 2 clinical trials, caused a dose-dependent reduction of AKT activation, at concentrations causing MMe cell growth arrest. In this study we firstly describe that MMe cells express aside from AKT1 also AKT3 and that either the myristoylated, constitutively active, forms of the two proteins, abrogated perifosine-mediated cell growth inhibition. Moreover, we describe here a novel mechanism of perifosine that interferes, upstream of AKT, affecting EGFR and MET phosphorylation. Finally, we demonstrate a significant increase in cell toxicity when MMe cells were treated with perifosine in combination with cisplatin.This study provides a novel mechanism of action of perifosine, directly inhibiting EGFR/MET-AKT1/3 axis, providing a rationale for a novel translational approach to the treatment of MMe

Electronic structure and lattice dynamics of 1T-VSe2_2: origin of the 3D-CDW

In order to characterize in detail the charge density wave (CDW) transition of 1$T$-VSe$_2$, its electronic structure and lattice dynamics are comprehensively studied by means of x-ray diffraction, angle resolved photoemission (ARPES), diffuse and inelastic x-ray scattering (IXS), and state-of-the-art first principles density functional theory calculations. Resonant elastic x-ray scattering (REXS) does not show any resonant enhancement at either V or Se K-edges, indicating that the CDW peak describes a purely structural modulation of the electronic ordering. ARPES identifies (i) a pseudogap at T$>$T$_{CDW}$, which leads to a depletion of the density of states in the $ML-M'L'$ plane at T$<$T$_{CDW}$, and (ii) anomalies in the electronic dispersion reflecting a sizable impact of phonons on it. A diffuse scattering precursor, characteristic of soft phonons, is observed at room temperature (RT) and leads to the full collapse of the low-energy phonon ($\omega_1$) with propagation vector (0.25 0 -0.3) r.l.u. We show that the frequency and linewidth of this mode are anisotropic in momentum space, reflecting the momentum dependence of the electron-phonon interaction (EPI), hence demonstrating that the origin of the CDW is, to a much larger extent, due to the momentum dependence EPI with a small contribution from nesting. The pressure dependence of the $\omega_1$ soft mode remains nearly constant up to 13 GPa at RT, with only a modest softening before the transition to the high-pressure monoclinic $C2/m$ phase. The wide set of experimental data are well captured by our state-of-the art first-principles anharmonic calculations with the inclusion of van der Waals (vdW) corrections in the exchange-correlation functional. The description of the electronics and dynamics of VSe$_2$ reported here adds important pieces of information to the understanding of the electronic modulations of TMDs