Nanoscale characterization of single Au nanorods by confocal microscopy

We analyzed the scattering patterns of individual Au nanorods detected by means of confocal interference scattering microscopy in combination with a higher order laser mode. We placed the Au nanorods at the interface between two dielectric media and examined the influence of different interfaces on the signal amplitude, the signal-to-noise ratio as well as on the precision of topological measurements. Approaching the index matching regime allows for topological measurements with high accuracy minimizing the acquisition time. We were also able to track the position and the orientation of particles embedded in water even when they were not thoroughly sticking to the glass surface. These results underscore the potential of the presented technique for applications in life sciences.     

Poly(glyceryl glycerol): A multi‐functional hydrophilic polymer for labeling with boronic acids

The synthesis of poly(glyceryl glycerol) (PGG), a polymer featuring a polyethylene oxide backbone and 1,2‐diol groups in every repeating unit, is presented. PGG was prepared by monomer‐activated ring‐opening polymerization of (dl ?1,2‐isopropylidene glyceryl) glycidyl ether, introducing a functional azido‐ or bromo‐head group to each chain. The 1,2‐diol groups, which were released by acidic deprotection, readily reacted with boronic acid derivatives, enabling the attachment of functional moieties under mild aqueous conditions. PGG was conjugated to poly(l ‐lactide) (PLLA) via azide‐alkyne cycloaddition and the resulting copolymer assembled into nanoparticles of 70 nm diameter in aqueous solution. Labeling of the PGG–PLLA particles was achieved by simple mixing with a boronic acid‐functional fluorophore. The labeling efficiency was determined by fluorescence spectroscopy to be 85.5% for boronic acid‐functional rhodamine B compared with 0.2% for plain rhodamine B. The strong interaction of PGG with boronic acids is ascribed to its polyol structure. This study demonstrates the usefulness and versatility of PGG as a hydrophilic polymer for possible biomedical applications. © 2017 Wiley Periodicals, Inc. J. Polym. Sci., Part A: Polym. Chem. 2017 , 55 , 1822–1830     

Comparison of beaked whale detection algorithms

Due to recent advances in passive acoustic monitoring techniques, beaked whales are now more effectively detected acoustically than visually during vessel-based (e.g. line-transect) surveys. Beaked whales signals can be discriminated from those of other cetaceans by the unique characteristics of their echolocation clicks (e.g. duration >175 μs, center frequencies between 30 and 40 kHz, inter-click intervals between 0.2 and 0.4 s and frequency upsweeps). Furthermore, these same characteristics make these signals ideal candidates for testing automated detection and classification algorithms. There are several different beaked whale automated detectors currently available for use. However, no comparative analysis of detectors exists. Therefore, comparison between studies and datasets is difficult. The purpose of this study was to test, validate, and compare algorithms for detection of beaked whales in acoustic line-transect survey data. Six different detection algorithms (XBAT, Ishmael, PAMGUARD, ERMA, GMM and FMCD) were evaluated and compared. Detection trials were run on three sample days of towed-hydrophone array recordings collected by NOAA Southwest Fisheries Science Center (SWFSC) during which were confirmed visual sightings of beaked whales (Ziphius cavirostris and Mesoplodon peruvianus). Detections also were compared to human verified acoustic detections for a subset of these data. In order to measure the probabilities of false detection, each detector was also run on three sample recordings containing clicks from another species: Risso’s dolphin (Grampus griseus). Qualitative and quantitative comparisons and the detection performance of the different algorithms are discussed.     

Proteomic analysis of Pseudomonas aeruginosa grown under magnesium limitation

In this study, large-scale qualitative and quantitative proteomic technology was applied to the analysis of the opportunistic bacterial pathogen Pseudomonas aeruginosa grown under magnesium limitation, an environmental condition previously shown to induce expression of various virulence factors. For quantitative analysis, whole cell and membrane proteins were differentially labeled with isotope-coded affinity tag (ICAT) reagents and ICAT reagent-labeled peptides were separated by two-dimensional chromatography prior to analysis by electrospray ionization-tandem mass spectrometry (ESI-MS/MS) in an ion trap mass spectrometer (ITMS). To increase the number of protein identifications, gas-phase fractionation (GPF) in the m/z dimension was employed for analysis of ICAT peptides derived from whole cell extracts. The experiments confirmed expression of 1331 P. aeruginosa proteins of which 145 were differentially expressed upon limitation of magnesium. A number of conserved Gram-negative magnesium stress-response proteins involved in bacterial virulence were among the most abundant proteins induced in low magnesium. Comparative ICAT analysis of membrane versus whole cell protein indicated that growth of P. aeruginosa in low magnesium resulted in altered subcellular compartmentalization of large enzyme complexes such as ribosomes. This result was confirmed by 2-D PAGE analysis of P. aeruginosa outer membrane proteins. This study shows that large-scale quantitative proteomic technology can be successfully applied to the analysis of whole bacteria and to the discovery of functionally relevant biologic phenotypes.     

On the syntheses of 8-Heteroaryl-substituted 9-(β-D-Ribofuranosyl)-2,6-diaminopurines through Pd-catalyzed coupling in the presence of cupric oxide

Convenient methods for the preparation of 9-(β-D-ribofuranosyl) derivatives of 8-(2- and 3-thienyl)-2,6-diaminopurine and of 8-(2- and 3-furyl)-2,6-diaminopurine, which are potential antiviral agents has been worked out. The key step was a Pd(0)-catalyzed Stille coupling between 2- and 3-tributylstannylthiophene and 2- and 3-tributylstannylfuran and trimethylsilyl protected 9-(β-D-ribofuranosyl)-2,6-diamino-8-bro-mopurine. The use of N,N-dimethylformamide as solvent at 110° and dichloro(diphenylphosphine-propane)palladium(II) [PdC1₂(dppp)] with cupric oxide as co-reagent was essential in order to obtain a fast reaction and high yields.     

Remarkably efficient removal of toxic bromate from drinking water with a porphyrin–viologen covalent organic framework

The presence of carcinogenic bromate (BrO₃⁻) in drinking water became a global concern and efforts towards its removal mainly focused on addressing the source. Herein, we rationally designed a porphyrin-based covalent organic framework (PV-COF) with a cationic surface to provide electrostatic interactions and a porphyrin core to induce hydrogen bonding interactions for the efficient removal of BrO₃⁻ from water. Through H-bonding and electrostatic interactions, PV-COF exhibited an exceptional bromate removal efficiency (maximum adsorption capacity, Q_max: 203.8 mg g⁻¹) with the fastest uptake rate (k_ads) of 191.45 g mg⁻¹ min⁻¹. The bromate concentration was reduced to far below the allowed concentration in drinking water (10 ppb) within 20 minutes. We studied the relationship between bromate adsorption and COF surface modification by metalation of the porphyrinic core or neutralization of the viologen linkers by chemical reduction. The bromate adsorption mechanism was studied by EDAX mapping and molecular simulations, and it was found that ion exchange and hydrogen bonding formation drive the adsorption. Importantly, PV-COF could be easily recycled several times without compromising its adsorption efficiency.

A cationic COF removes carcinogenic bromate with a remarkable rate constant of 191.45 g mg⁻¹ min⁻¹.     

Preparation of analyte-sensitive polymeric supports for biochemical sensors

The preparation and use of multiple polymers attached to a surface plasmon resonance (SPR) sensor for optimization of signal enhancement and minimization of fouling during sensing of biological species has been achieved. These polymers are advantageous compared to the current practice of carboxymethylated-dextran (CM-dextran). The polymers offer a wide range of functionalities and different molecular weights. Using these polymers, the SPR sensors can be fabricated as fast or faster than the CM-dextran sensor. In this study, we investigated the use of nine polymers for SPR biosensors. Polysaccharides, including CM-dextran, CM-hyaluronic acid, hyaluronic acid, and alginic acid, were investigated. Humic acid, polylactic acid, polyacrylic acid, orthopyridyldisulfide-polyethyleneglycol-N-hydroxysuccinimide (OPSS-PEG-NHS) and a synthesized polymer; polymethacrylic-acid-co-vinyl-acetate (PMAVA), were also used. The polymers were chemically attached to a thiol monolayer on the SPR biosensor using carbodiimide chemistry. The polymers were functionalized for binding of anti-myoglobin (anti-MG). The sensor performance was measured using myoglobin (MG) at 25 ng ml⁻¹, a biologically relevant level for myocardial infarction detection. Most polymers offered similar performance to CM-dextran for MG detection in HEPES buffer saline pH 7.4 (HBS). In preliminary studies in bovine serum, each of the candidate polymers demonstrated better performance than CM-dextran.     

Capillary electrophoretic and micellar electrokinetic separations of asymmetric dimethyl-L-arginine and structurally related amino acids: quantitation in human plasma

We report the development of efficient electrophoretic methods for the separation and quantification of L-arginine and six naturally occurring derivatives that are structurally and functionally related. Capillary electrophoresis (CE) employing a concentrated borate buffer at pH 9.4 achieves the separation of mixtures containing dimethyl-L-arginine, NG-monomethyl-L-arginine, L-arginine, L-homoarginine, L-ornithine, and L-citrulline as 4-fluoro-7-nitrobenzofurazan derivatives. In addition, the separation of the isomeric dimethyl-L-arginine derivatives (symmetric and asymmetric) is attained with baseline resolution by micellar electrokinetic chromatography (MEKC) when a high concentration of deoxycholic acid is added as a surfactant to the same running buffer. The influence of buffer type, concentration, and pH on the separation was studied to optimize separation conditions. The limit of quantitation (LOQ) for asymmetric dimethyl-L-arginine in aqueous solution was determined to be 20 microM using UV absorption in a CE separation and 0.1 microM using laser induced fluorescence (LIF) detection in an MEKC separation. This newly developed method was successfully applied for the quantitation of asymmetric dimethyl-L-arginine and L-arginine in human plasma samples at levels that might be used as a clinical diagnostic for cardiovascular disease (0.125 microM LOQ).     

193-nm Photodissociation of ions from saturated and unsaturated aliphatic molecules

To determine the analytical utility of photodissociation as a general fragmentation technique for tandem mass spectrometry of organic ions, the ability to fragment those ions considered least likely to absorb photons efficiently was investigated. To this end, the ability to photodissociate ions of aliphatic compounds by using 193-nm photons has been studied. Three fragment ions, the C₄H ₉ ⁺ ion from n-hexane, the C₄H ₇ ⁺ ion from 2-hexene, and C₄H ₅ ⁺ from 2-hexyne, have been photodissociated. The fragmentation efficiencies for all three ions studied were between 25 and 45%. The photofragment ion spectrum for each precursor ion studied is made up of characteristic fragments. These spectra demonstrate the ability to photodissociate aliphatic ions that originate from both saturated and unsaturated molecules. This provides substantial hope that virtually all organic ions will be able to be photodissociated by using 193-nm photons.