Monitoring metabolites consumption and secretion in cultured cells using ultra-performance liquid chromatography quadrupole–time of flight mass spectrometry (UPLC–Q–ToF-MS)

Here we present an ultra-performance liquid chromatography–mass spectrometry (UPLC–MS) method for extracellular measurements of known and unexpected metabolites in parallel. The method was developed by testing 86 metabolites, including amino acids, organic acids, sugars, purines, pyrimidines, vitamins, and nucleosides, that can be resolved by combining chromatographic and m/z dimensions. Subsequently, a targeted quantitative method was developed for 80 metabolites. The presented method combines a UPLC approach using hydrophilic interaction liquid chromatography (HILIC) and MS detection achieved by a hybrid quadrupole–time of flight (Q–ToF) mass spectrometer. The optimal setup was achieved by evaluating reproducibility and repeatability of the analytical platforms using pooled quality control samples to minimize the drift in instrumental performance over time. Then, the method was validated by analyzing extracellular metabolites from acute lymphoblastic leukemia cell lines (MOLT-4 and CCRF-CEM) treated with direct (A-769662) and indirect (AICAR) AMP activated kinase (AMPK) activators, monitoring uptake and secretion of the targeted compound over time. This analysis pointed towards a perturbed purine and pyrimidine catabolism upon AICAR treatment. Our data suggest that the method presented can be used for qualitative and quantitative analysis of extracellular metabolites and it is suitable for routine applications such as in vitro drug screening.     

Electrochemical milling and faceting: size reduction and catalytic activation of palladium nanoparticles

Improved performance through milling: A method for enhancing the catalytic activity of supported metal nanoparticles is reported. This method enhances the activity for the ethanol electro-oxidation of a supported palladium catalyst. The much higher catalytic performance is ascribed to the increased electrochemically active surface area as well as the generation of high-index facets at the milled nanoparticle surface.     

Experimental electron density of sumanene, a bowl-shaped fullerene fragment; comparison with the related corannulene hydrocarbon

The experimental electron density of sumanene, C(21)H(12), was extracted from a high resolution X-ray data set measured at 100 K and topologically analyzed. In addition to bond topological and atomic properties, information about the density distribution between adjacent molecules, which show close C···C approaches of ~3.4 ? within the columnar π-stacks in the crystal lattice, are discussed. A comparison is made with the electron density of the related corannulene molecule based also on the analysis of Electron Localizability Indicator (ELI-D) calculations.     

Development and validation of a robust capillary electrophoresis method for impurity profiling of etomidate including the determination of chiral purity using a dual cyclodextrin system

An enantioselective CE assay for the simultaneous determination of the enantiomeric purity and of related substances of etomidate has been developed and validated using a binary chiral selector system employing 30 mg/mL beta-CD and 4.6 mg/mL sulfated-beta-CD in a 150 mM potassium phosphate buffer, pH 2.1. The method was validated with respect to specificity, range, linearity, LOQ and LOD, precision and accuracy. The assay allowed the detection and determination of related substances including (S)-etomidate at the 0.05% w/w level, the reporting threshold as defined by the International Conference on Harmonisation guidelines as well as the European Pharmacopoeia. Robustness testing was carried out by an "Augmented Plackett-Burman" design. Quantitation of the compounds was performed by calibration graphs with respect to lidocaine hydrochloride as internal standard and by peak area normalization, the procedure usually applied by pharmacopoeias. Although data obtained from the calibration graphs constructed with the aid of the internal standard were more accurate based on compound recovery, peak area normalization may also be used without significant loss of accuracy and precision.     

Solid-phase synthesis of terminal oligonucleotide-phosphoramidate conjugates

A novel phosphoramidite, N,N-diisopropylamino-2-cyanoethyl-9-anthracenemethyl phosphoramidite 1, was prepared and coupled with the terminal 5′-hydroxyl of support-bound T₁₀ and the putative phosphite triester intermediate was subsequently reacted with iodine in the presence of either water or a series of primary and secondary amines. The reactivity of 1 compared to a previously reported benzyl phosphoramidite 2 was also investigated: oxidation of the product of coupling 2 with CPG-T₁₀-5′OH under aqueous conditions resulted in greater than 30% of the benzyl moiety being retained. In contrast, essentially complete loss of the 9-anthracenemethyl group was observed using 1 under the same conditions. Oligonucleotides modified with a terminal phosphate monoester, lipophilic, fluorescent or cationic groups were thus prepared.     

Backward stochastic differential equations with jumps and related non-linear expectations

In this paper, we are interested in real-valued backward stochastic differential equations with jumps together with their applications to non-linear expectations. The notion of non-linear expectations has been studied only when the underlying filtration is given by a Brownian motion and in this work the filtration will be generated by both a Brownian motion and a Poisson random measure. We study at first backward stochastic differential equations driven by a Brownian motion and a Poisson random measure and then introduce the notions of f$f$-expectations and of non-linear expectations in this set-up.     

Modelling and optimization of least-cost water distribution networks with multiple supply sources and users

The proper allocation of water resources is a very important practical problem in the field of water network planning. Optimization models that are expeditious and easy to use for all stakeholders of the sector play an important role for water resource management. The present work resumes and reviews a least-cost optimization model proposed by our group (Maiolo and Pantusa in Water Sci Tech-W Sup.  https://doi.org/10.2166/ws.2015.114, 2016), able to design a water distribution network with multiple supply sources and multiple users. This approach requires of solving an optimization problem based on a nonlinear objective function which is proportional to the cost of the water distribution network. The cost of pre-existing pipelines is considered null. A more realistic scenario, able to consider the maximum flow rate allowed for existing sources-users connections, is considered here. In order to illustrate the usefulness and flexibility of the proposed approach, an application of the model to the real case of the province of Croton, Southern Italy, is presented.     

Erbium- and chlorine-doped fluorozirconate-based glasses for up-converted fluorescence

A series of glass ceramics was prepared and the structure–property relationships evaluated. The base composition of the series was comprised of fluorozirconate (FZ) and fluorochlorozirconate (FCZ) glass ceramics. Thermal processing of as-made FCZ-based glass ceramics produced sharp peaks in X-ray diffraction patterns, which was attributed to BaCl₂ nanocrystals formed in the material; this was not observed in the FZ glass. Up-conversion fluorescence experiments were carried out for both the FZ and FCZ samples doped with erbium. The fact that an increase in the up-conversion efficiency was not observed indicates that erbium ions were not embedded in the BaCl₂ nanocrystals. A comparison is made between this series and a similar series that was doped with neodymium.     

The thermal study of oxyfluoride glass with strontium fluoride

A thulium (Tm 3+) doped oxyfluoride glass ceramic containing SrF2 nanocrystals has been presented. Transparent glass ceramic was obtained by heat treating the glass from the SiO2-Al2O3-ZnO-SrF2 system at the first crystallization temperature. Cerammization of glass was studied by DTA/DSC, XRD and SEM methods. It has been found that nanocrystallization of SrF2 strongly depends on the ratio between the components and amount of SrF2. Moreover the rare earth dopant Tm3+ influenced on the thermal properties of glass. The formation of SrF2 nanocrystals in glass ceramic was confirmed by X-ray diffraction. X-ray diffraction analysis of the transparent glass ceramic revealed that the SrF2 nanocrystals are precipitated in the glass matrix. Analysis of the local atomic interactions in the structure of oxyfluoride glass has been used to explain the course of the crystallization.