Screening an In-House Isoquinoline Alkaloids Library for New Blockers of Voltage-Gated Na+ Channels Using Voltage Sensor Fluorescent Probes: Hits and Biases

Voltage-gated Na⁺ (Na_V) channels are significant therapeutic targets for the treatment of cardiac and neurological disorders, thus promoting the search for novel Na_V channel ligands. With the objective of discovering new blockers of Na_V channel ligands, we screened an In-House vegetal alkaloid library using fluorescence cell-based assays. We screened 62 isoquinoline alkaloids (IA) for their ability to decrease the FRET signal of voltage sensor probes (VSP), which were induced by the activation of Na_V channels with batrachotoxin (BTX) in GH3b6 cells. This led to the selection of five IA: liriodenine, oxostephanine, thalmiculine, protopine, and bebeerine, inhibiting the BTX-induced VSP signal with micromolar IC₅₀. These five alkaloids were then assayed using the Na⁺ fluorescent probe ANG-2 and the patch-clamp technique. Only oxostephanine and liriodenine were able to inhibit the BTX-induced ANG-2 signal in HEK293-hNa_V1.3 cells. Indeed, liriodenine and oxostephanine decreased the effects of BTX on Na⁺ currents elicited by the hNa_V1.3 channel, suggesting that conformation change induced by BTX binding could induce a bias in fluorescent assays. However, among the five IA selected in the VSP assay, only bebeerine exhibited strong inhibitory effects against Na⁺ currents elicited by the hNav1.2 and hNav1.6 channels, with IC₅₀ values below 10 µM. So far, bebeerine is the first BBIQ to have been reported to block Na_V channels, with promising therapeutical applications.     

The Ameliorative Effect of Empagliflozin in Vigabatrin-Induced Cerebellar/Neurobehavioral Deficits: Targeting mTOR/AMPK/SIRT-1 Signaling Pathways

Introduction. Vigabatrin (VGB) is an antiepileptic drug that acts to irreversibly inhibit the γ-aminobutyric acid (GABA) transaminase enzyme, elevating GABA levels. Broad studies have established that long-term treatment and/or high doses of VGB lead to variable visual defects. However, little attention has been paid to its other side effects, especially those demonstrating cerebellar involvement. Sodium glucose-linked co-transporter 2 (SGLT2) inhibitors are antidiabetic agents with protective effects far greater than expected based on their anti-hyperglycemic effect. Method. Our study herein was designed to investigate the possible ameliorative effect of empagliflozin, the SGLT2 inhibitors, in VGB-induced cerebellar toxicity. A total of 40 male Wistar rats were allocated equally into 4 groups: Group I: control group; Group II: VGB group; Group III empagliflozin treated VGB group; and Group IV: empagliflozin treated group. All groups were subjected to the detection of cerebellar messenger RNA gene expression of silent mating type information regulation 2 homolog 1 (SIRT1) and Nucleoporin p62 (P62). Mammalian target of rapamycin (mTOR), adenosine monophosphate-activated protein kinase (AMPK), and beclin1 levels were assessed by the ELISA technique while malondialdehyde (MDA) level and superoxide dismutase (SOD) activity were detected spectrophotometrically. Immuno-histochemical studies, focusing on glial fibrillary acidic protein (GFAP) and S100 were performed, and the optical color density and the mean area percentage of GFAP positive astrocytes and the number of S 100 positive cells were also counted. Results. Following empagliflozin treatment, we documented significant upregulation of both SIRT1 and P62 mRNA gene expression. Additionally, AMPK, Beclin1 levels, and SOD activity were significantly improved, while both mTOR and MDA levels were significantly reduced. Conclusions. We concluded for the first time that empagliflozin efficiently ameliorated the VGB-induced disrupted mTOR/AMPK/SIRT-1 signaling axis with subsequent improvement of the autophagy machinery and mitigation of the oxidative and inflammatory cellular environment, paving the way for an innovative therapeutic potential in managing VGB-induced neurotoxicity.     

Polyfluorene Thin Films Synthesized by a Novel Plasma Polymerization Method

The synthesis of polyfluorene (PF) thin films by simultaneously superposing a continuous and pulsed discharge and the characterizations of these samples are presented. The double discharge plasma system is constructed by superposing two discharges; namely, a low pressure dc glow one and a high current pulsed one. The fluorene monomer in powder form was vaporized in the system at argon plasma without any modification, at 0.5 mbar operating pressure. The structure of the thin films was investigated via XPS, UV–visible, FTIR, XRD and SEM. The FTIR and the UV–visible results revealed that the fluorene structure was retained at the produced samples. Semi-conducting behavior was established, and upon the iodine doping, the optical energy band gap (E _g ) dropped down from 3.7 to 2.4 eV. The morphology of the synthesized PF thin films was amorphous, with granular structures of different sizes depending on the location of the substrate.     

Thermal stability and UV–Vis-NIR spectroscopy of a new erbium-doped fluorotellurite glass

A new transparent bulk glass from the system 76TeO₂?·?10ZnO?·?9.0PbO?·?1.0PbF₂?·?3.0Na₂O doped with Er³⁺ (TZPPN doped with Er³⁺) has been prepared using the conventional melt-quenching method. Results of differential thermal analysis (DTA) measurements indicate good thermal stability of this glass. The refractive indices at different wavelengths, the optical energy gap, the Sellmeier gap energy and the dispersion energy have been estimated. The Judd–Ofelt parameters, Ω _t (t?=?2,?4,?6) of Er³⁺ were evaluated from optical absorption spectra. Electric dipole, magnetic dipole type transition probabilities, spectroscopic quality factors, branching ratio and radiative lifetimes of several excited states of Er³⁺ have been predicted using intensity Judd–Ofelt parameters. The spectroscopic properties indicate that TZPPN glass doped with Er³⁺ is a promising candidate for laser applications and may be suitable for upconversion fibre optical devices.     

The improvement of InAs/GaAs quantum dot properties capped by Graphene

InAs self‐assembled quantum dots (QDs) were grown by molecular beam epitaxy on (001) GaAs substrate. Uncapped and capped QDs with GaAs and graphene layers were studied using atomic force microscopy and Raman spectroscopy. Graphene multi‐layer was grown by chemical vapor deposition and transferred on InAs/GaAs QDs. It is well known that the presence of a cap layer modifies the size, shape, and density of the QDs. According to the atomic force microscopy study, in contrast to the GaAs capped sample, which induce a dramatic decrease of the density and height of dots, graphene cap layer sample presents a slight influence on the surface morphology and the density of the islands compared with the uncapped one. The difference shown in the Raman spectra of the samples is due to change of strain and alloy disorder effects on the QDs. Residuals strain and the relaxation coefficients have been investigated. All results confirm the best crystalline quality of the graphene cap layer dots sample relative to the GaAs capped one. So graphene can be used to replace GaAs in capping InAs/GaAs dots. To our knowledge, such study has not been carried out until now. Copyright © 2013 John Wiley & Sons, Ltd.     

Ultrafast X‐ray imaging of laser–metal additive manufacturing processes

The high‐speed synchrotron X‐ray imaging technique was synchronized with a custom‐built laser‐melting setup to capture the dynamics of laser powder‐bed fusion processes in situ. Various significant phenomena, including vapor‐depression and melt‐pool dynamics and powder‐spatter ejection, were captured with high spatial and temporal resolution. Imaging frame rates of up to 10 MHz were used to capture the rapid changes in these highly dynamic phenomena. At the same time, relatively slow frame rates were employed to capture large‐scale changes during the process. This experimental platform will be vital in the further understanding of laser additive manufacturing processes and will be particularly helpful in guiding efforts to reduce or eliminate microstructural defects in additively manufactured parts.     

Radiolabelling of TiO<Subscript>2</Subscript> nanoparticles for radiotracer studies

Industrially manufactured titanium dioxide nanoparticles have been successfully radiolabelled with ⁴⁸V by irradiation with a cyclotron-generated proton beam. Centrifugation tests showed that the ⁴⁸V radiolabels were stably bound within the nanoparticle structure in an aqueous medium, while X-ray diffraction indicated that no major structural modifications to the nanoparticles resulted from the proton irradiation. In vitro tests of the uptake of cold and radiolabelled nanoparticles using the human cell line Calu-3 showed no significant difference in the uptake between both batches of nanoparticles. The uptake was quantified by Inductively Coupled Plasma Mass Spectrometry and high resolution γ-ray spectrometry for cold and radiolabelled nanoparticles, respectively. These preliminary results indicate that alterations to the nanoparticles’ properties introduced by proton bombardment can be controlled to a sufficient extent that their further use as radiotracers for biological investigations can be envisaged and elaborated.     

Numerical and experimental studies of molten pool formation during an interaction of a pulse laser (Nd:YAG) with a magnesium alloy

A pulse laser (Nd:YAG) interaction with an AZ91 magnesium alloy has been experimentally and numerically studied. A two-dimensional (2D) axisymmetric model of a molten pool created by a laser heat source has been developed. The elaborated model solves the coupled equations of a laminar fluid flow and heat transfer to demonstrate the flow behavior in the pool. This model takes into account the coupled effects of buoyancy and Marangoni forces, the thermophysic variation properties with temperature, and the radiation and convection heat losses. Concerning numerical results, the molten temperature distribution, velocity field and molten shape were discussed. It was noted that the Marangoni flow significantly alters the characteristics of the thawing and solidifying processes, and makes the molten pool wider and shallower. On the other hand, the experimental results showed that the material thermal properties have significant effects on the transport phenomena which takes place in the molten pool, and consequently on the formation as well as the shape of the pool. Finally, a comparison between the numerical and experimental results exhibited a good agreement.