Genetic, genomic and physiological state studies on single-needle bio-electrosprayed human cells

Bio-electrospraying, a non-contact jet-based direct cell engineering approach, was recently pioneered and demonstrated for handling a wide range of primary living cells. In those studies, post-treated cells were biologically assessed in comparison to several controls by way of flow cytometry. Although flow cytometry accurately assesses those viable populations of cells, subtle effects at a sub-cellular level could have been missed. Therefore, in the present study we demonstrate metaphase chromosome breakage studies carried out on single-needle bio-electrosprayed human T-lymphocytes, which are compared with several controls. The results indicate that post-treated T-lymphocytes do not demonstrate any increase in chromosome damage in comparison to control cells. These studies further validate bio-electrospraying as a technique with potential for clinical utility.     

Nickel‐Catalyzed Esterification of Aliphatic Amides

Recent studies have demonstrated that amides can be used in nickel‐catalyzed reactions that lead to cleavage of the amide C?N bond, with formation of a C?C or C?heteroatom bond. However, the general scope of these methodologies has been restricted to amides where the carbonyl is directly attached to an arene or heteroarene. We now report the nickel‐catalyzed esterification of amides derived from aliphatic carboxylic acids. The transformation requires only a slight excess of the alcohol nucleophile and is tolerant of heterocycles, substrates with epimerizable stereocenters, and sterically congested coupling partners. Moreover, a series of amide competition experiments establish selectivity principles that will aid future synthetic design. These studies overcome a critical limitation of current Ni‐catalyzed amide couplings and are expected to further stimulate the use of amides as synthetic building blocks in C?N bond cleavage processes.     

Neoglycopeptide dendrimer libraries as a source of lectin binding ligands

[structure: see text] A 15 625-membered peptide dendrimer combinatorial library was acylated with an alpha-C-fucosyl residue at its four N-termini and screened for binding to fucose-specific lectins. Dendrimer FD2 (Fuc-alpha-CH2CO-Lys-Pro-Leu)4(Lys-Phe-Lys-Ile)2Lys-His-Ile-NH2 was identified as a potent ligand against Ulex europaeus lectin UEA-I (IC50 = 11 microM) and Pseudomonas aeruginosa lectin PA-IIL (IC50 = 0.14 microM).     

General method for ultrathin free-standing films of nanofibrous composite materials

A simple and general method for the preparation of ultrathin, large-area, free-standing films of nanofibrous composite materials was developed. First, positively charged cadmium hydroxide nanostrands of 1.9 nm in diameter and micrometers in length were prepared by mixing dilute aqueous solutions of cadmium chloride and aminoethanol. Negatively charged dye molecules, proteins, nanoparticles, and water-soluble fullerene or carbon nanotubes were mixed with the nanostrands to give a well-dispersed solution of the corresponding nanofibrous composites. After filtration of the dispersions with a polycarbonate membrane filter, uniform films with a thickness of a few tens to hundreds of nanometers and a diameter of a few centimeters were obtained. The films were readily peeled off from the membrane filter by immersion in ethanol. The resultant free-standing films could be further transferred to other substrates, such as quartz plate, gold electrode, and porous alumina membrane, and were characterized by scanning and transmission electron microscopies. We provide herein various nanofibrous free-standing films with optical, biological, metallic, and magnetic properties.     

Determination of reducing ends with flow injection analysis with amperometric detection: application to enzyme-hydrolysed methyl cellulose

A novel method for detection of reducing ends of sugars is proposed, based on the use of as the oxidant in combination with amperometric detection and flow injection analysis (FIA). The method is very sensitive, giving values of <10 μM for the limit of detection for a series of mono- and oligosaccharides. Samples can be analysed every 30 s, and injection can be made fully automated, making it possible to perform on-line analysis of polysaccharide samples subjected to hydrolysis. Three methylcelluloses (MC) of different qualities were hydrolysed with three different glucanases, and the concentrations of reducing ends prior to, during and after hydrolysis were determined. Differences were observed between the results obtained using different combinations of enzymes and MCs, which revealed different selectivities of the various enzymes for the different substrates. One MC was also hydrolysed and analysed in real-time for three hours. The method proposed is superior to many of the standard methods used today, which require manual labour and have a lower sensitivity. Figure Set-up used for the instrumentation in the FIA system with automated injection. A pump delivers the reaction solution to the autosampler, where the samples are injected; the sample and solution react in a temperature-controlled random coil and the response is detected using an amperometric detection cell     

Conformational analysis of lipid molecules by self-organizing maps

The authors have studied the use of the self-organizing map (SOM) in the analysis of lipid conformations produced by atomic-scale molecular dynamics simulations. First, focusing on the methodological aspects, they have systematically studied how the SOM can be employed in the analysis of lipid conformations in a controlled and reliable fashion. For this purpose, they have used a previously reported 50 ns atomistic molecular dynamics simulation of a 1-palmitoyl-2-linoeayl-sn-glycero-3-phosphatidylcholine (PLPC) lipid bilayer and analyzed separately the conformations of the headgroup and the glycerol regions, as well as the diunsaturated fatty acid chain. They have elucidated the effect of training parameters on the quality of the results, as well as the effect of the size of the SOM. It turns out that the main conformational states of each region in the molecule are easily distinguished together with a variety of other typical structural features. As a second topic, the authors applied the SOM to the PLPC data to demonstrate how it can be used in the analysis that goes beyond the standard methods commonly used to study the structure and dynamics of lipid membranes. Overall, the results suggest that the SOM method provides a relatively simple and robust tool for quickly gaining a qualitative understanding of the most important features of the conformations of the system, without a priori knowledge. It seems plausible that the insight given by the SOM could be applied to a variety of biomolecular systems and the design of coarse-grained models for these systems.     

A method of test for residual isophorone diisocyanate trimer in new polyester-polyurethane coatings on light metal packaging using liquid chromatography with tandem mass spectrometric detection

A method of test for residual isophorone diisocyanate (IPDI) trimer in experimental formulation polyester-polyurethane (PEPU) thermoset coatings on metal food packaging is described. The method involves extraction of coated panels using acetonitrile containing dibutylamine for concurrent derivatisation, and then high performance liquid chromatography with electrospray ionisation tandem mass spectrometric detection (LC-MS/MS). Single laboratory validation was carried out using three different experimental PEPU-based coatings. The calibrations were linear, the analytical recovery was good, no interferences were seen, and substance identification criteria were met. The detection limit of the method is around 0.02 micro g/100 cm(2) of coating, which for a typical sized can and assuming complete migration of any residual IPDI trimer, corresponds to about 0.2 micro g/kg food or beverage. Separate studies indicated that, even if migration occurred at such low levels, the IPDI trimer would not be expected to persist in canned aqueous or fatty foodstuffs as it would hydrolyse to the corresponding aliphatic amine or react with food components to destroy the isocyanate moiety. The method of test developed here for residual IPDI trimer in thermoset polyester-polyurethane coatings should prove to be a valuable tool for investigating the cure kinetics of these novel coatings and help to guide the development of enhanced formulations.     

Asymmetric alpha-alkynylation of piperidine via N-sulfinyliminium salts

Piperidine was stereoselectively alpha-alkynylated in a four-step sequence made up of transformation to a chiral nonracemic N-sulfinylpiperidine, anodic oxidation to N-sulfinyliminium ion equivalent, alkynylation through addition of a mixed organoaluminum derivative, and final acidic deprotection of the sulfoxide. Overall yields are around 50%, and the diastereoselectivity of the nucleophilc addition was between 92 and 99% de, allowing isolation of the final product with 99% enantiomeric purity.