Light-Responsive Elastin-Like Peptide-Based Targeted Nanoparticles for Enhanced Spheroid Penetration

We describe here a near infrared light-responsive elastin-like peptide (ELP)-based targeted nanoparticle (NP) that can rapidly switch its size from 120 to 25 nm upon photo-irradiation. Interestingly, the targeting function, which is crucial for effective cargo delivery, is preserved after transformation. The NPs are assembled from (targeted) diblock ELP micelles encapsulating photosensitizer TT1-monoblock ELP conjugates. Methionine residues in this monoblock are photo-oxidized by singlet oxygen generated from TT1, turning the ELPs hydrophilic and thus trigger NP dissociation. Phenylalanine residues from the diblocks then interact with TT1 via π-π stacking, inducing the re-formation of smaller NPs. Due to their small size and targeting function, the NPs penetrate deeper in spheroids and kill cancer cells more efficiently compared to the larger ones. This work could contribute to the design of “smart” nanomedicines with deeper penetration capacity for effective anticancer therapies.     

Promoter-Controlled Synthesis and Conformational Analysis of Cyclic Mannosides up to a 32-mer

Cyclodextrins are widely used as carriers of small molecules for drug delivery owing to their remarkable host properties and excellent biocompatibility. However, cyclic oligosaccharides with different sizes and shapes are limited. Cycloglycosylation of ultra-large bifunctional saccharide precursors is challenging due to the constrained conformational spaces. Herein we report a promoter-controlled cycloglycosylation approach for the synthesis of cyclic α-(1→6)-linked mannosides up to a 32-mer. Cycloglycosylation of the bifunctional thioglycosides and (Z)-ynenoates was found to be highly dependent on the promoters. In particular, a sufficient amount of a gold(I) complex played a key role in the proper preorganization of the ultra-large cyclic transition state, providing a cyclic 32-mer polymannoside, which represents the largest synthetic cyclic polysaccharide to date. NMR experiments and a computational study revealed that the cyclic 2-mer, 4-mer, 8-mer, 16-mer, and 32-mer mannosides adopted different conformational states and shapes.     

An environmental friendly method for the automatic determination of hypochlorite in commercial products using multisyringe flow injection analysis

A multisyringe flow injection analysis system was used for the determination of hypochlorite in cleaning agents, by measurement of the native absorbance of hypochlorite at 292 nm. The methodology was based on the selective decomposition of hypochlorite by a cobalt oxide catalyst giving chloride and oxygen. The difference of the absorbance of the sample before and after its pass through a cobalt oxide column was selected as analytical signal. As no further reagent was required this work can be considered as a contribution to environmental friendly analytical chemistry. The entire analytical procedure, including in-line sample dilution in three steps was automated by first, dilution in a stirred miniature vessel, second by dispersion and third by in-line addition of water using multisyringe flow injection technique. The dynamic concentration range was 0.04-0.78 g L⁻¹ (relative standard deviation lower than 3%), where the extension of the hypochlorite decomposition was of 90 ± 4%. The proposed method was successfully applied to the analysis of commercial cleaning products. The accuracy of the method was established by iodometric titration.     

Tumor Carbohydrate Associated Antigen Analogs as Potential Binders for Siglec-7

We investigated two recently synthesized and characterized sialyl derivatives, bearing the Neu5Ac-α-(2-6)-Gal epitope, as promising binders for Siglec-7, an inhibitory Siglec mainly found on natural killer cells. A variety of sialoglycan structures can be recognized by Siglec-7 with implications in the modulation of immune responses. Notably, overexpression of sialylated glycans recognized by Siglec-7 can be associated with the progression of several tumors, including melanoma and renal cell carcinoma. NOE-based NMR techniques, including Saturation Transfer Difference and transferred-NOESY NMR, together with molecular docking and dynamic simulations were combined to shed light on the molecular basis of Siglec-7 recognition of two conformationally constrained Sialyl-Tn antigen analogs. We, therefore, identify the ligands epitope mapping and their conformational features and propose 3D models accurately describing the protein-ligand complexes. We found that the binding site of Siglec-7 can accommodate both synthetic analogs, with the sialic acid mainly involved in the interaction. Moreover, the flexibility of Siglec-7 loops allows a preferred accommodation of the more rigid compound bearing a biphenyl moiety at position 9 of the sialic acid that contributed to the interaction to a large extent. Our findings provided insights for developing potential novel high affinity ligands for Siglec-7 to hinder tumor evasion.     

Using biased randomization for solving the two-dimensional loading vehicle routing problem with heterogeneous fleet

This paper discusses the two-dimensional loading capacitated vehicle routing problem (2L-CVRP) with heterogeneous fleet (2L-HFVRP). The 2L-CVRP can be found in many real-life situations related to the transportation of voluminous items where two-dimensional packing restrictions have to be considered, e.g.: transportation of heavy machinery, forklifts, professional cleaning equipment, etc. Here, we also consider a heterogeneous fleet of vehicles, comprising units of different capacities, sizes and fixed/variable costs. Despite the fact that heterogeneous fleets are quite ubiquitous in real-life scenarios, there is a lack of publications in the literature discussing the 2L-HFVRP. In particular, to the best of our knowledge no previous work discusses the non-oriented 2L-HFVRP, in which items are allowed to be rotated during the truck-loading process. After describing and motivating the problem, a literature review on related work is performed. Then, a multi-start algorithm based on biased randomization of routing and packing heuristics is proposed. A set of computational experiments contribute to illustrate the scope of our approach, as well as to show its efficiency.     

Simple and rapid determination of biogenic amines in wine by liquid chromatography-electrospray ionization ion trap mass spectrometry

A rapid liquid chromatographic-electrospray ionisation ion trap mass spectrometry (LC-ESI-ITMS) method has been developed for the routine analysis of eight of the most oenologically important biogenic amines in wine without any sample pre-treatment. The method involves addition of heptylamine as an internal standard (IS) and the direct injection of filtered wine samples previously diluted with ultra high purity (UHP) water. The full-scan MS-MS spectra and the identical retention times to those of reference standards were used for unequivocal identification of the analytes. For most amines, the most abundant ions were derived from the loss of an ammonia group, while in the case of spermine and the I.S. the major product ions arose from the loss of 1,3-propyldiamine and the production of adduct with water, respectively. Detection was achieved in positive ionisation with an ion trap mass spectrometer operating in multiple-reaction monitoring (MRM) mode. The method allowed accurate determination of the analytes in the range 0.5-40 ng mL⁻¹. Within-day and between-day relative standard deviation percentages were <8% and <12%, respectively. The overall process was successfully applied to identify and quantify biogenic amines in Rioja red wines. The new method is sensitive, rapid, cheap and less labour intensive.     

The York map as a Shanmugadhasan canonical transformation in tetrad gravity and the role of non-inertial frames in the geometrical view of the gravitational field

A new parametrization of the 3-metric allows to find explicitly a York map by means of a partial Shanmugadhasan canonical transformation in canonical ADM tetrad gravity. This allows to identify the two pairs of physical tidal degrees of freedom (the Dirac observables of the gravitational field have to be built in term of them) and 14 gauge variables. These gauge quantities, whose role in describing generalized inertial effects is clarified, are all configurational except one, the York time, i.e. the trace of the extrinsic curvature of the instantaneous 3-spaces (corresponding to a clock synchronization convention) of a non-inertial frame centered on an arbitrary observer. In the Dirac Hamiltonian is the sum of the weak ADM energy (whose density is coordinate-dependent, containing the inertial potentials) and of the first-class constraints. The main results of the paper, deriving from a coherent use of constraint theory, are: (i) The explicit form of the Hamilton equations for the two tidal degrees of freedom of the gravitational field in an arbitrary gauge: a deterministic evolution can be defined only in a completely fixed gauge, i.e. in a non-inertial frame with its pattern of inertial forces. The simplest such gauge is the 3-orthogonal one, but other gauges are discussed and the Hamiltonian interpretation of the harmonic gauges is given. This frame-dependence derives from the geometrical view of the gravitational field and is lost when the theory is reduced to a linear spin 2 field on a background space-time. (ii) A general solution of the super-momentum constraints, which shows the existence of a generalized Gribov ambiguity associated to the 3-diffeomorphism gauge group. It influences: (a) the explicit form of the solution of the super-momentum constraint and then of the Dirac Hamiltonian; (b) the determination of the shift functions and then of the lapse one. (iii) The dependence of the Hamilton equations for the two pairs of dynamical gravitational degrees of freedom (the generalized tidal effects) and for the matter, written in a completely fixed 3-orthogonal Schwinger time gauge, upon the gauge variable , determining the convention of clock synchronization. The associated relativistic inertial effects, absent in Newtonian gravity and implying inertial forces changing from attractive to repulsive in regions with different sign of , are completely unexplored and may have astrophysical relevance in the interpretation of the dark side of the universe.     

Bioactive Betalain Extracts from Cactus Pear Fruit Pulp,Beetroot Tubers,and Amaranth Leaves

Natural food items and the additional benefits they provide have received considerable attention in recent years. Betalains are nutritious pigments which have valuable biological properties, e.g., antioxidant and free radical scavenging activity. In this study, aqueous betalain extracts were obtained from different coloured cactus pears (purple, red/pink, and orange), amaranth, and beetroot, with and without the addition of ascorbic acid, microwave-heated, and freeze-dried and subsequently analysed by thin layer chromatography (TLC). Beetroot samples without the addition of ascorbic acid (AA) had lower phenols, flavonoids, and ascorbic acid content than beetroot samples extracted with the addition of AA. Amaranth had significantly higher contents of antioxidants than all the other plants. Results for phenolic compounds showed that there were no significant differences between cactus pear cultivars, however, significant differences were seen between the two beetroot samples (microwave-heated with and without AA) as well as amaranth. For flavonoid compounds, amaranth had significantly higher values than all other samples. The lowest flavonoid content was found in beetroot without AA (0.49 mgCE/g). For ascorbic acid, significant differences were noticed between amaranth (71.71 mg/100 g) and samples from cactus pear and beetroot. TLC results showed that purple and red cactus pear samples had the most vivid colours, a reflection of the high betacyanin and betaxanthin contents in the cultivars. Moreover, extracts from cactus pear, beetroot, and amaranth were classified according to a decision tree which was designed by the Code of Federal Regulations/Food Additives Regulation of the EU. The classification of betalain pigment extracts as colouring foods was achieved through enrichment factor calculations and the colourant decision tree. The results showed that the betalain pigment extraction method used is inexpensive, time-saving, energy-saving, non-toxic, and chemical solvent free and yields high concentrations of betalains.     

A mild oxidative method for the preparation of γ-hydroxy-α-nitroolefins from α,β-epoxyketoximes using IBX

An efficient method for the preparation of γ-hydroxy-α-nitroolefins from α,β-epoxyketoximes has been developed using IBX. The reaction occurs at room temperature without the formation of over-oxidation products and also has an easy work-up procedure.     

Identification and quantification of glucosinolates in rapeseed using liquid chromatography–ion trap mass spectrometry

A rapid and sensitive method for the speciation and quantification of glucosinolates in rapeseed is described. The method combines liquid chromatography (LC) with ion trap mass spectrometry (ITMS) detection. Electrospray ionization (ESI) has been chosen as the ionization technique for the on-line coupling of LC with ITMS. Glucosinolates are extracted from different rapeseeds with MeOH and the extracts are cleaned-up by solid phase extraction with Florisil cartridges. Aqueous extracts are injected into LC system coupled to an ITMS, leading to accurately quantify eight of the most important glucosinolates in rapeseed, by MS² mode and confirming their structure by MS³ acquisition. All the glucosinolates found in rapeseeds provide good signals corresponding to the deprotonated precursor ion [M-H]⁻. The method is reliable and reproducible, and detection limits range from 0.5 nmol g⁻¹ to 3.7 nmol g⁻¹ when 200 mg of dried seeds of certified reference material are analyzed. Within-day and between-day RSD percentages range between 2.4–14.1% and 3.9–16.9%, respectively. The LC-ESI-ITMS-MS method described here allows for a rapid assessment of these metabolites in rapeseed without a desulfatation step. The overall process has been successfully applied to identify and quantify glucosinolates in rapeseed samples.