The Cellular Interactions of PEGylated Gold Nanoparticles: Effect of PEGylation on Cellular Uptake and Cytotoxicity

Poly(ethylene glycol) (PEG) is frequently used to coat various medical nanoparticles (NPs). As PEG is known to minimize NP interactions with biological specimens, the question remains whether PEGylated NPs are intrinsically less toxic or whether this is caused by reduced NP uptake. In the present work, the effect of gold NP PEGylation on uptake by three cell types is compared and evaluated the effect on cell viability, oxidative stress, cell morphology, and functionality using a multiparametric methodology. The data reveal that PEGylation affects cellular NP uptake in a cell‐type‐dependent manner and influences toxicity by different mechanisms. At similar intracellular NP numbers, PEGylated NPs are found to yield higher levels of cell death, mostly by induction of oxidative stress. These findings reveal that PEGylation significantly reduces NP uptake, but that at similar functional (= cell‐associated) NP levels, non‐PEGylated NPs are better tolerated by the cells.     

Benign reduction of carbon nanotube agglomerates using a supercritical carbon dioxide process

A method was developed to deagglomerate commercially available multi-walled carbon nanotube (MWCNT) bundles while maintaining the carbon nanotube aspect ratio. The process utilizes the rapid expansion of a supercritical carbon dioxide/MWCNT mixture to separate large primary carbon nanotube agglomerates. High levels of deagglomeration of Baytubes^® C 150 P and Nanocyl? NC-7000 MWCNT bundles were observed on the macroscale and nanoscale, resulting in 30-fold and 50-fold decreases in bulk density, respectively, with median agglomerate sizes <8 μm in diameter. These results were obtained while retaining the aspect ratio of the as-received nanomaterial, irrespective of the MWCNT agglomerate morphology. It was found that a temperature and pressure of 40 °C and 7.86 MP resulted in maximum deagglomeration without damage to the MWCNTs. Thermodynamic principles were applied to describe the effect of processing variables on the efficiency of the deagglomeration. These results suggest that combining this process with a composite processing step, such as melt compounding, will result in nanocomposites with enhanced electrical properties.     

Isomeric discrimination of arginine-containing dipeptides using electrospray ionization-ion trap mass spectrometry and the kinetic method

The Kinetic Method (KM), applied commonly for thermochemical determinations, is used here for sterically-controlled isomeric determination of N- versus C-terminal Arg-containing dipeptide isomers (ArgX versus XArg; where X = His, Leu, Lys, Pro, Ser, Phe, and Tyr). The KM is offered as an alternate approach to direct collision-induced dissociation (CID) procedures. Through formation, isolation, and dissociation of a sterically-encumbered, metal-centered complex with electrospray ionization ion trap mass spectrometry technology, reference dipeptide molecules are screened to quantitatively differentiate a mixture of isomers based on their arrangement about the metal center. Arg-containing dipeptide molecules are chosen because of their contribution in a wide array of protein and peptide functions. Additionally, problems cited previously for evaluation of systems containing Arg residues (due to the incorporation of the guanidinium moiety) by the KM are addressed. The method is shown to be successful for highlighting favorable reference analytes (e.g., ArgPhe, ArgLeu, ProArg, PheArg, among others) for exceptional discrimination (R(iso) > 2.0) of the majority of N- and C-terminal Arg-containing peptides tested.     

High throughput screening of library compounds against an oligonucleotide substructure of an RNA target

Approximately 300,000 compounds from selected libraries were screened against a subdomain of a hepatitis C viral (HCV) RNA using a high throughput flow injection mass spectrometry (FIA-MS) method with automated data storage and analysis. Samples contained 2 microM RNA target and 10 microM of each of up to ten ligands. Preliminary studies to optimize operational parameters used the binding of aminoglycosides to the A44 subdomain of bacterial RNA. Binding (confirmed by titration) and sensitivity were maximized within the constraints of the library and throughput. The mobile phase of 5 mM ammonium acetate in 50% isopropanol maintained the noncovalent complexes and provided good detection by electrospray mass spectrometry. Additionally, this composition maximized general solubility of the various classes of compounds including the oligonucleotide and organic library molecules. Cation adduction was insignificant in this screen although some solute and target dependent acetate adduction was observed. The ion trap mass spectrometer provided sufficient mass resolution to identify complexes of RNA with known components of the library. Converted mass spectral data (netCDF) were subjected to two types of statistical evaluation based on binding. The first algorithm identified noncovalent complexes that correlated with the molecular weights of the injected compounds. The second yielded the largest peak in the noncovalent complex region of the spectrum; this spectrum may or may not correlate with expected well components. Sixty-three compounds were confirmed to bind by more stringent secondary testing. Titrations, which were carried out with selected binding compounds, yielded a range of dissociation constants. Biological activity was observed for eleven confirmed binders.     

Quantitative determination of the peptide retention of polymeric substrates using matrix-assisted laser desorption/ionization mass spectrometry

Polymer surface-peptide binding interactions have been shown previously to lead to reductions in peptide matrix assisted laser desorption/ionization (MALDI) ion signals. In previous studies, increases in surface-peptide binding were characterized by the increases in both the initially adsorbed and retained quantities of 125I-radiolabeled peptides. The present studies establish a specific correlation between the peptide retention properties of the polymer surface and the reduction in the peptide MALDI ion signal. This correlation is demonstrated by obtaining MALDI mass spectra of angiotensin I applied to various polymer surfaces having a range of peptide adsorption and retention properties. In addition, the use of a MALDI based method of standard additions is shown to allow the quantitation of the polymer surface-peptide retention affinity for angiotensin I and porcine insulin. The MALDI standard additions method for measurement of surface-peptide retention affinities offers a number of significant advantages over conventional radiolabeled peptide binding methods and promises to be a valuable tool for the determination of this important biomaterial characteristic.     

A combined solid-state NMR and X-ray crystallography study of the bromide ion environments in triphenylphosphonium bromides

Multinuclear ((31)P and (79/81)Br), multifield (9.4, 11.75, and 21.1 T) solid-state nuclear magnetic resonance experiments are performed for seven phosphonium bromides bearing the triphenylphosphonium cation, a molecular scaffold found in many applications in chemistry. This is undertaken to fully characterise their bromine electric field gradient (EFG) tensors, as well as the chemical shift (CS) tensors of both the halogen and the phosphorus nuclei, providing a rare and novel insight into the local electronic environments surrounding them. New crystal structures, obtained from single-crystal X-ray diffraction, are reported for six compounds to aid in the interpretation of the NMR data. Among them is a new structure of BrPPh(4), because the previously reported one was inconsistent with our magnetic resonance data, thereby demonstrating how NMR data of non-standard nuclei can correct or improve X-ray diffraction data. Our results indicate that, despite sizable quadrupolar interactions, (79/81)Br magnetic resonance spectroscopy is a powerful characterisation tool that allows for the differentiation between chemically similar bromine sites, as shown through the range in the characteristic NMR parameters. (35/37)Cl solid-state NMR data, obtained for an analogous phosphonium chloride sample, provide insight into the relationship between unit cell volume, nuclear quadrupolar coupling constants, and Sternheimer antishielding factors. The experimental findings are complemented by gauge-including projector-augmented wave (GIPAW) DFT calculations, which substantiate our experimentally determined strong dependence of the largest component of the bromine CS tensor, δ(11), on the shortest Br-P distance in the crystal structure, a finding that has possible application in the field of NMR crystallography. This trend is explained in terms of Ramsey's theory on paramagnetic shielding. Overall, this work demonstrates how careful NMR studies of underexploited exotic nuclides, such as (79/81)Br, can afford insights into structure and bonding environments in the solid state.     

A macrocyclic β-iodoallenolate intermediate is key: synthesis of the ABD core of phomactin A

An enantioselective strategy for the synthesis of phomactin natural products is described. The Lewis acid triggered cyclization of a β-iodoallenolate embedded in a 12-membered macrocycle was used to obtain a highly functionalized bicyclo[9.3.1]pentadecane in good yield and high diastereoselectivity. This iodoenone contains the substituents of the AD ring system of the phomactin family of natural products, appropriate for further functionalization. Synthesis of the oxadecalin core of phomactin A from the AD iodoenone intermediate was achieved. In this unusual strategy, rings A and B are both fashioned within a macrocyclic precursor.     

Generation of [(IPr)Pd(PR2Cl)] complexes via P-Cl reductive elimination

Treatment of [(IPr)Pd(Cl)(2)(PR(2)H)] (IPr = 1,3-bis(2,6-diisopropylphenyl)imidazol-2-ylidene; R = Cy, tBu, or 1-Ad) with NaN(SiMe(3))(2) generated isolable [(IPr)Pd(PR(2)Cl)] complexes (68-75%) that have been crystallographically characterized. The formation of these mixed-ligand Pd(0) species in this manner corresponds to an unusual net dehydrohalogenation/P-Cl reductive elimination sequence.     

A Luminescent and Sublimable Dy(III) -Based Single-Molecule Magnet

The reaction of [Ln(hfac)(3) ]?2?H(2) O and pyridine-N-oxide (PyNO) leads to isostructural dimers of the formula [Ln(hfac)(3) (PyNO)](2) (Ln=Eu, Gd, Tb, Dy). The Dy derivative shows a remarkable single-molecule magnet behavior with complex hysteresis at 1.4?K. The dynamics of the magnetization features are two relaxation regimes: a thermally activated one at high temperature (τ(0) =(5.62±0.4)×10(-11) s and Δ=(167±1)?K) and a quantum tunneling regime at low temperature with a tunneling frequency of 0.42?Hz. The analysis of the Gd derivative evidences intradimer antiferromagnetic interactions (J=(-0.034±0.001)?cm(-1) ). Moreover, the Eu, Tb, and Dy derivatives are luminescent with quantum yield of 51, 53, and 0.1?%, respectively. The thermal investigation of [Dy(hfac)(3) (PyNO)](2) shows that the dimers can be sublimated intact, suggesting their possible exploit as active materials for surface-confined nanostructures to be investigated by fluorimetry methods.     

Two‐Photon Absorption and Time‐Resolved Stimulated Emission Depletion Spectroscopy of a New Fluorenyl Derivative

The synthesis, comprehensive linear photophysical characterization, two‐photon absorption (2PA), steady‐state and time‐resolved stimulated emission depletion properties of a new fluorene derivative, (E)‐1‐(2‐(di‐p‐tolylamino)‐9,9‐diethyl‐9H‐fluoren‐7‐yl)‐3‐(thiophen‐2‐yl)prop‐2‐en‐1‐one ( 1 ), are reported. The primary linear spectral properties, including excitation anisotropy, fluorescence lifetimes, and photostability, were investigated in a number of aprotic solvents at room temperature. The degenerate 2PA spectra of 1 were obtained with open‐aperture Z‐scan and two‐photon induced fluorescence methods, using a 1 kHz femtosecond laser system, and maximum 2PA cross‐sections of ～400–600 GM were obtained. The nature of the electronic absorption processes in 1 was investigated by DFT‐based quantum chemical methods implemented in the Gaussian 09 program. The one‐ and two‐photon stimulated emission spectra of 1 were measured over a broad spectral range using a femtosecond pump–probe‐based fluorescence quenching technique, while a new methodology for time‐resolved fluorescence emission spectroscopy is proposed. An effective application of 1 in fluorescence bioimaging was demonstrated by means of one‐ and two‐photon fluorescence microscopy images of HCT 116 cells containing dye encapsulated micelles.