Ultrasensitive proteomics using high-efficiency on-line micro-SPE-nanoLC-nanoESI MS and MS/MS

Ultrasensitive nanoscale proteomics approaches for characterizing proteins from complex proteomic samples of <50 ng of total mass are described. Protein identifications from 0.5 pg of whole proteome extracts were enabled by ultrahigh sensitivity (<75 zmol for individual proteins) achieved using high-efficiency (peak capacities of approximately 10(3)) 15-microm-i.d. capillary liquid chromatography separations (i.e., using nanoLC, approximately 20 nL/min mobile-phase flow rate at the optimal linear velocity of approximately 0.2 cm/s) coupled on-line with a micro-solid-phase sample extraction and a nanoscale electrospray ionization interface to a 11.4-T Fourier transform ion cyclotron resonance (FTICR) mass spectrometer (MS). Proteome measurement coverage improved as sample size was increased from as little as 0.5 pg of sample. It was found that a 2.5-ng sample provided 14% coverage of all annotated open reading frames for the microorganism Deinococcus radiodurans, consistent with previous results for a specific culture condition. The estimated detection dynamic range for detected proteins was 10(5)-10(6). An improved accurate mass and LC elution time two-dimensional data analysis methodology, used to both speed and increase the confidence of peptide/protein identifications, enabled identification of 872 proteins/run from a single 3-h nanoLC/FTICR MS analysis. The low-zeptomole-level sensitivity provides a basis for extending proteomics studies to smaller cell populations and potentially to a single mammalian cell. Application with ion trap MS/MS instrumentation allowed protein identification from 50 pg (total mass) of proteomic samples (i.e., approximately 100 times larger than FTICR MS), corresponding to a sensitivity of approximately 7 amol for individual proteins. Compared with single-stage FTICR measurements, ion trap MS/MS provided a much lower proteome measurement coverage and dynamic range for a given analysis time and sample quantity.     

Electrochemical oxidation of adsorbed alkenoic acids as a function of chain length at Pt(111) electrodes. Studies by cyclic voltammetry,EELS and Auger spectroscopy

Studies of the electrochemical oxidation of a series of straight-chain terminal alkenoic acids adsorbed at a Pt(111) electrode surface are reported. Compounds adsorbed were: propenoic acid (acrylic acid, PPA); 3-butenoic acid (vinylacetic acid, 3BTA); 4-pentenoic acid (allylacetic acid, 4PTA); 6-heptenoic acid (6HPA); and 10-undecenoic acid (10UDA). Vibrational spectra of adsorbed layers were obtained by use of electron energy-loss spectroscopy (EELS). Molecular packing densities were measured by use of Auger spectroscopy. Electrochemical oxidation of each adsorbed layer was explored by means of cyclic voltammetry in aqueous inert electrolyte (KF/HF). As the analogous aliphatic acids are not chemisorbed at Pt under the same conditions, the alkenoic acids evidently adsorb at Pt(111) predominantly through the C=C double bond. Molecular packing densities indicate that the carboxylic acid moiety is in contact with the Pt surface only in the case of PPA. EELS spectra also indicate that the carboxylate groups (other than in PPA) are present as pendants. The carboxylic acid O-H stretching bands of most of the adsorbed acids are red-shifted and broadened, evidently due to extensive intermolecular hydrogen bonding; the exceptions are PPA, for which the interaction is primarily with the Pt surface, and 3BTA, for which intermolecular interaction between the carboxylic acid pendants is apparently prevented by steric considerations. The surface-attached carboxylic acid moieties react with KOH solution, leading to retention of K⁺ ions, detected by Auger spectroscopy, and to changes in the vibrational spectra indicative of carboxylate anions; reactivity toward KOH decreases with chain length. Adsorbed alkenoic acids at Pt(111) surfaces are stable in water and in vacuum. Oxidation of the adsorbed short-chain acids PPA and 3BTA proceeds to completion, forming CO₂ as the principal product. Oxidation of the adsorbed long-chain acids converts the C=C moiety to 2CO₂, and transforms the remainder of the molecule to an unadsorbed diacid (likely possibilities are malonic acid from 4PTA; glutaric acid from 6HPA; and heptane-1,7-dioic acid from 10UDA).     

Functionality of chitosan-halloysite nanocomposite films for sustained delivery of antibiotics: The effect of chitosan molar mass

This study was designed to investigate functionality of tetracycline-loaded chitosan-halloysite nanocomposite films, with focus on evaluating the influence of chitosan molar mass on films applicability for sustained local antibiotic delivery. The films were prepared by casting and solvent evaporation using low, medium, and high molar mass chitosan. SEM analysis revealed compact, nonporous and rough surface of the nanocomposite films due to the presence of halloysite agglomerates and tetracycline crystals. Increasing chitosan molar mass led to higher values of elongation at break (from 21.65 ± 2.65 to 34.48 ± 2.34%), tensile strength (from 134.8 ± 13.21 to 246.36 ± 14.69 MPa), and elastic modulus (from 633.79 ± 128.37 to 716.55 ± 60.76 MPa) of the nanocomposite films. FT-IR, XRPD, and thermal analyses confirmed molar mass dependent chitosan-halloysite interactions and improved thermal stability of the nanocomposite films in comparison with chitosan films. The nanocomposite films released tetracycline in a sustained manner, with the slowest release achieved from the films consisting of low molar mass chitosan. Chitosan molar mass was confirmed to be a functionality-related characteristic of chitosan-halloysite nanocomposite films as potential sustained-release carriers for topical delivery of antibiotics. © 2019 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2020 , 137, 48406.     

Properties of silver nanostructure-coated PTFE and its biocompatibility

Silver nanolayers were sputtered on polytetrafluoroethylene (PTFE) and subsequently transformed into discrete nanoislands by thermal annealing. The Ag/PTFE composites prepared under different conditions were characterized by several complementary methods (goniometry, UV-visible spectroscopy, X-ray photoelectron spectroscopy, and atomic force microscopy), and new data on the mechanism of Ag layer growth and Ag atom clustering under annealing were obtained. Biocompatibility of selected Ag/PTFE composites was studied in vitro using vascular smooth muscle cell (VSMC) cultures. Despite of the well-known inhibitory properties of silver nanostructures towards broad spectrum of bacterial strains and cells, it was found that very thin silver coating stimulates both adhesion and proliferation of VSMCs.