Liposomal FRET Assay Identifies Potent Drug-Like Inhibitors of the Ceramide Transport Protein (CERT)

Ceramide transfer protein (CERT) mediates non-vesicular transfer of ceramide from endoplasmic reticulum to Golgi apparatus and thus catalyzes the rate-limiting step of sphingomyelin biosynthesis. Usually, CERT ligands are evaluated in tedious binding assays or non-homogenous transfer assays using radiolabeled ceramides. Herein, a facile and sensitive assay for CERT, based on Förster resonance energy transfer (FRET), is presented. To this end, we mixed donor and acceptor vesicles, each containing a different fluorescent ceramide species. By CERT-mediated transfer of fluorescent ceramide, a FRET system was established, which allows readout in 96-well plate format, despite the high hydrophobicity of the components. Screening of a 2 000 compound library resulted in two new potent CERT inhibitors. One is approved for use in humans and one is approved for use in animals. Evaluation of cellular activity by quantitative mass spectrometry and confocal microscopy showed inhibition of ceramide trafficking and sphingomyelin biosynthesis.     

Catalyst-free direct synthesis of β-enaminones through reaction of benzohydrazonamides with cyclic 1,3-diketones: Access to exocyclic β-enaminones

Abstract

The reaction of benzohydrazonamides with cyclic 1,3-diketones affords β-enaminones in good to excellent yields (14 examples, 69???89%). This transformation proceeds under mild reaction conditions (ethyl acetate, 25?°C, 12???24?h) in the absence of any catalyst and gives access to novel nitrogen-rich organic scaffolds.     

Synthesis,characterization, antimicrobial activity, 3D-QSAR,DFT, and molecular docking of some ciprofloxacin derivatives and their copper(II) complexes

Six new derivatives of ciprofloxacin compounds and their copper(II) complexes were synthesized, characterized by spectroscopic methods (ultraviolet–visible [UV–vis], Fourier transform infrared [FTIR], nuclear magnetic resonance [NMR], mass spectrometry [MS], and electron paramagnetic resonance [EPR]), and tested for antibacterial activities against gram-negative and gram-positive bacteria. The data showed that ciprofloxacin derivatives act as bidentate ligands and the metal ions coordinate through the pyridone carbonyl and the carboxylate oxygen atoms. Tetragonally distorted octahedral ligand fields were assumed for all complexes based on their spectral studies. Copper(II) complexes of the synthesized ciprofloxacin derivatives revealed higher antibacterial activities against gram-positive and gram-negative bacterial species than the parent ciprofloxacin antibiotic. Furthermore, three-dimensional quantitative structure–activity relationship (3D-QSAR) models were evaluated by studying 30 antibiotic compounds of the quinolone class. Density function theory (DFT) calculations were applied to evaluate the optimized geometrical structures using the B3LYP method and 6-311G(d,p) basis set. The 3D-QSAR study revealed that there are eight optimum parameters that give the best predictive modulation with good reliability (R² = 0.996, F = 12.004, sigma = 0.426). In silico molecular docking was also performed on the derivatives, and the results revealed the presence of two types of interactions between the Escherichia coli and the derivatives, H-bonding and Van der Waals interactions, and an effective inhibition at the docked site.     

Mass Distributions and Charge Dispersion for the Nuclear Fission

The theory of nuclear fission is reconsidered by introducing the charge asymmetry in the asymmetric two center shell model as a dynamical collective coordinate. The quantum mechanical fluctuations, which are accompanied with the collective motion as a function of the mass asymmetry, are responsible for the mass distributions in nuclear fission. Numerical calculations are carried out for the mass distributions and charge dispersions for the nuclear fission of the ²³⁶U and ²³⁸U nuclei. The present obtained theoretical calculations are in good agreement with the experimental measurements.     

Collective mass parameters in the nuclear fission process

The theory of nuclear fission process is considered. The effective mass parameters are calculated as a function of the following collective coordinates: the separation between the centres of harmonic oscillators, the mass asymmetry parameter and the necking parameter. Numerical calculations are carried out for the fission of²³⁶U and²³⁸U nuclei. Symmetric and asymmetric cases are considered. From the present calculation we see the importance of taking into account the necking parameter as a dynamical collective coordinate.     

Two new cytotoxic furoquinoline alkaloids isolated from Aegle marmelos (Linn.) Correa

Two new cytotoxic furoquinoline alkaloids were isolated from the leaves of Aegle marmelos (Linn.) Correa; one from the total alkaloidal fraction (acid/base shake-out method) of the CHCl₃ extract and identified as 7,8-dihydroxy-4-hydrofuroquinoline and named trivially as Aegelbine-A. The other new alkaloid isolated from the pet. ether extract and identified as 4-hydro-7-hydroxy-8-prenyloxyfuroquinoline and named trivially as Aegelbine-B, together with a known alkaloid; aegeline and a known phenolic acid; ρ-hydroxybenzoic acid. The structures of all the isolated compounds were established based on 1D and 2D NMR spectroscopy and HR-ESI/MS. The cytotoxic activity of the isolated compounds was evaluated in vitro against HepG-2, PC3, A549 and MCF-7 cell lines. The obtained results revealed promising activity with structure-based relationship which is discussed briefly.     

Effect of electrolytes on the retention behavior of some benzenesulfonates in electrochemically modulated liquid chromatography

The effects of electrolytes on the retention behavior of some benzenesulfonates in electrochemically modulated liquid chromatography were studied. Both cations and anions were found to have considerable effects on retention. As cation size increases, retention decreases, while anions show more complicated effects were anionic size and charge distribution contribute to the overall behavior of anions. Large anions with a delocalized negative charge on the whole species result in lower retention times, and vice versa. Also, electrolyte concentration plays an important role in the retention behavior observed. Initially, as electrolyte concentration was increased retention increased due to electrostatic interactions of cations with the negatively charged stationary phase. However, retention starts to slightly decrease or increase after some specific electrolyte concentration depending on the nature of the electrolytic species. Finally, an interesting behavior of double peak appearance of a single solute was observed at low electrolyte concentrations and was attributed to the presence of other active sites on the carbon stationary phase.     

Synthesis,DFT Calculations,Antiproliferative, Bactericidal Activity and Molecular Docking of Novel Mixed-Ligand Salen/8-Hydroxyquinoline Metal Complexes

Despite the common use of salens and hydroxyquinolines as therapeutic and bioactive agents, their metal complexes are still under development. Here, we report the synthesis of novel mixed-ligand metal complexes (MSQ) comprising salen (S), derived from (2,2′-{1,2-ethanediylbis[nitrilo(E) methylylidene]}diphenol, and 8-hydroxyquinoline (Q) with Co(II), Ni(II), Cd(II), Al(III), and La(III). The structures and properties of these MSQ metal complexes were investigated using molar conductivity, melting point, FTIR, ¹H NMR, ¹³C NMR, UV–VIS, mass spectra, and thermal analysis. Quantum calculation, analytical, and experimental measurements seem to suggest the proposed structure of the compounds and its uncommon monobasic tridentate binding mode of salen via phenolic oxygen, azomethine group, and the NH group. The general molecular formula of MSQ metal complexes is [M(S)(Q)(H₂O)] for M (II) = Co, Ni, and Cd or [M(S)(Q)(Cl)] and [M(S)(Q)(H₂O)]Cl for M(III) = La and Al, respectively. Importantly, all prepared metal complexes were evaluated for their antimicrobial and anticancer activities. The metal complexes exhibited high cytotoxic potency against human breast cancer (MDA-MB231) and liver cancer (Hep-G2) cell lines. Among all MSQ metal complexes, CoSQ and LaSQ produced IC₅₀ values (1.49 and 1.95 µM, respectively) that were comparable to that of cisplatin (1.55 µM) against Hep-G2 cells, whereas CdSQ and LaSQ had best potency against MDA-MB231 with IC₅₀ values of 1.95 and 1.43 µM, respectively. Furthermore, the metal complexes exhibited significant antimicrobial activities against a wide spectrum of both Gram-positive and -negative bacterial and fungal strains. The antibacterial and antifungal efficacies for the MSQ metal complexes, the free S and Q ligands, and the standard drugs gentamycin and ketoconazole decreased in the order AlSQ > LaSQ > CdSQ > gentamycin > NiSQ > CoSQ > Q > S for antibacterial activity, and for antifungal activity followed the trend of LaSQ > AlSQ > CdSQ > ketoconazole > NiSQ > CoSQ > Q > S. Molecular docking studies were performed to investigate the binding of the synthesized compounds with breast cancer oxidoreductase (PDB ID: 3HB5). According to the data obtained, the most probable coordination geometry is octahedral for all the metal complexes. The molecular and electronic structures of the metal complexes were optimized theoretically, and their quantum chemical parameters were calculated. PXRD results for the Cd(II) and La(III) metal complexes indicated that they were crystalline in nature.     

Spectrophotometric determination of enzymatically generated hydrogen peroxide using Sol-Gel immobilized horseradish peroxidase

Peroxidase entrapment in different Sol-Gel matrices was successful. The enzyme did not show a decrease in activity for at least 2 months as well as storage at room temperature and dry condition for periods exceeding 3 weeks. It was evident that the enzymatic activity was a function in the type of the alkoxysilane precursor. In addition, the optimum temperature which resulted in maximum enzymatic activity was also dependent on the type of Sol-Gel matrix. Excellent results were obtained for the determination of glucose in serum samples using soluble glucose oxidase in conjunction with the Sol-Gel entrapped peroxidase. The enzymatically produced hydrogen peroxide is oxidized by the entrapped peroxidase yielding oxygen which oxidizes the faint blue variamine blue into the intensely violet colored species (the molar absorptivity is about 1.8 x 10(4) 1 mol(-1) cm(-1)). The characteristics of this chromogenic system as well as optimized conditions for its use in the spectrophotometric determination of enzymatically generated hydrogen peroxide were investigated. Excellent agreement between the results obtained by the proposed method and the widely used standard method, utilizing a commercial reagents kit, was always observed.     

Three dimensional texture analysis in MRI: a preliminary evaluation in gliomas

The discrimination of tumor boundaries from normal tissue, as well as the evaluation of tissue heterogeneity and tumor grading often continue to pose a challenge in MRI. Although yielding promising results in various fields of medical imaging, two- dimensional (2D) texture analysis in MRI has, until now, demonstrated a lack of specificity in brain tumor classification. A new three-dimensional (3D) approach using Cooccurrence Matrix analysis is proposed to increase the sensitivity and specificity of brain tumor characterization. A preliminary comparative evaluation of 2D and 3D texture analysis was performed on T(1)-weighted MRI of seven gliomas for characterization of solid tumor, necrosis, edema and surrounding white matter. With 3D compared to 2D method, a better discrimination is obtained between necrosis and solid tumor as well as between edema and solid tumor. Using both methods, peritumoral white matter overlaps with edema, but is completely separated from far homo-lateral matter. This latter shows a complete overlapping with contra-lateral matter. The 3D texture analysis approach could provide a new tool for tumor grading and treatment follow-up, as well as for surgery or radiation therapy planning.