An Expanded Set of Fluorogenic Sulfatase Activity Probes

Fluorogenic probes that are activated by an enzymatic transformation are ideally suited for profiling enzyme activities in biological systems. Here, we describe two fluorogenic enzyme probes, 3‐O‐methylfluorescein‐sulfate and resorufin‐sulfate, that can be used to detect sulfatases in mycobacterial lysates. Both probes were validated with a set of commercial sulfatases and used to reveal species‐specific sulfatase banding patterns in a gel‐resolved assay of mycobacterial lysates. The fluorogenic probes described here are suitable for various assays and provide a starting point for creating new sulfatase probes with improved selectivity for mycobacterial sulfatases.     

Large Particle Penetration During PM10 Sampling

The objective of the present study was to characterize the performance of a federal reference method (FRM) PM₁₀ size-selective inlet using analysis methods designed to minimize uncertainty in measured sampling efficiencies for large particles such as those most often emitted from agricultural operations. The performance of an FRM PM₁₀ inlet was characterized in a wind tunnel at a wind speed of 8 km/h. Data were also collected for 20 and 25 μm particles at wind speeds of 2 and 24 km/h. Results of the present sampler evaluation compared well with those of previous studies for a similar inlet near the cutpoint, and the sampler passed the criteria required for certification as a FRM sampler when tested at 8 km/h. Sampling effectiveness values for particles with nominal diameters of 20 and 25 μm exceeded 3% for 8 and 24 km/h wind speeds in the present study and were statistically higher than both the “ideal” PM₁₀ sampler (as defined in 40 CFR 53) and the ISO (1995) standard definition of thoracic particles (p < 0.05) for 25 μm particles leading to the potential for significant sampling bias relative to the “ideal” PM₁₀ sampler when measuring large aerosols.

Copyright 2014 American Association for Aerosol Research     

Colourful questions of an Indian village

How we understand and recall colour in the environments we encounter is reliant on the context. Drawing on a repeated experience of the author in a rural Indian village, a number of questions were raised regarding visual distinctiveness and its impact on the identity of place embedded in the memories of the village. A photographic walk‐through was undertaken to determine the existing colour palette and its relationship to the character of the memories of the village. Combined with observations and theory drawn from the literature, it is proposed that considering the experience from a pluralist perspective enables insights to emerge. In this case, colour moments and colour contrast are key attributes of memories and identity construction for the village visitor.     

The Polymerization of Metal-Containing Vinylic Monomers of Iron and Tungsten with Metal–Carbon σ Bonds

A series of metal-containing vinylic monomers of the type and was homopolymerized using 2,2-azobisisobutyronitrile (AIBN) as the free-radical initiator. These monomers were also copolymerized with styrene in the presence of AIBN. These compounds represent a class of organometallic polymers in which the metal is bonded to the polymer backbone via a metal–carbon bond. The new compounds were characterized by IR and ¹H NMR spectroscopy as well as scanning electron microscopy, gel permeation chromatography, and thermoanalytical studies (DSC and TGA). The properties of the new organometallic polymers are discussed.     

Tape Casting of Lanthanum Chromite

The effects of process additives, ball milling, and solids loading were evaluated for tape casting suspensions of glycinenitrate-synthesized La_0.7Ca_0.31CrO₃ powder. An optimized formulation was obtained based on rheological characterization, electrokinetic sonic amplitude measurements, qualitative examination of green tapes, and the sintered microstructure. The tape casting formulation incorporated 66:34 methyl ethyl ketone/ethyl alcohol solvent, an aliphatic phosphate ester dispersant, and 80 wt% (35 vol%) solids. The best binder/plasticizer system was 12 wt% (15 vol%) poly(isobutyl methacrylate) and 5 wt% (6.3 vol%) benzyl butyl phthalate plasticizer (binder:plasticizer = 2.3). Cast tapes were sintered at 1300°C for 2 h, producing a bulk density of 96.2% theoretical, with linear shrinkage of 22% and an approximate grain size of 1.3 μm.     

Subcellular Localisation of a Quinoline-Containing Fluorescent Cyclometallated IrIII Complex in Plasmodium falciparum

A fluorescent analogue of a previously synthesised N,N-chelated Ir^III complex was prepared by coordination of the organic ligand to an extrinsic bis(2-phenylpyridine)iridium(III) fluorophore. This cyclometallated Ir^III complex in itself displays good, micromolar activity against the chloroquine-sensitive NF54 strain of Plasmodium falciparum. Live-cell confocal microscopy found negligible localisation of the fluorescent complex within the digestive vacuole of the parasite. This eliminated the haem detoxification pathway as a potential mechanism of action. Similarly, no localisation of the complex within the parasitic nucleus was found, thus suggesting that this complex probably does not interfere with the DNA replication process. A substantial saturation of fluorescence from the complex was found near phospholipid structures such as the plasma and nuclear membranes but not in neutral lipid bodies. This indicates that an association with these membranes, or organelles such as the endoplasmic reticulum or branched mitochondrion, could be essential to the efficacies of these types of antimalarial compounds.     

Pyrazole-Thiazole Core-Containing Analogs Exhibit Adjunctive Activity with Meropenem against Carbapenem-Resistant Enterobacteriaceae (CRE)

Pyrazole-thiazole core-containing compound KP-40 and 20 novel derivatives were designed and synthesized through traditional SAR analysis. These molecules displayed adjunctive activity with meropenem against Gram-negative bacteria evidenced by a range of fractional inhibitory concentration (FIC=0.5–0.25) and minimum adjunctive concentration (MAC=128–32 μM) values. Of this series of molecules, four compounds displayed notable adjunctive potential, with FIC and MAC values of 0.25 and 32 μM, respectively. Moreover, the solubility of these compounds was improved to an acceptable range. Further analysis using our “in house” permeation and efflux multi parameter optimization (PEMPO) algorithm revealed key physicochemical properties that may be critical for the development of active Gram-negative antibacterials. Taking PEMPO scores into consideration prior to executing synthesis of analogs may be a simple, yet rapid and effective strategy that can be used in conjunction with traditional SAR approaches to aid in the design of potent Gram-negative antibacterials.     

The effect of fertilizer placement on nitrogen uptake and yield of wheat and maize in Chinese loess soils

Field trials were carried out to study the fate of¹⁵N-labelled urea applied to summer maize and winter wheat in loess soils in Shaanxi Province, north-west China. In the maize experiment, nitrogen was applied at rates of 0 or 210 kg N ha^–1, either as a surface application, mixed uniformly with the top 0.15 m of soil, or placed in holes 0.1 m deep adjacent to each plant and then covered with soil. In the wheat experiment, nitrogen was applied at rates of 0, 75 or 150 kg N ha^–1, either to the surface, or incorporated by mixing with the top 0.15 m, or placed in a band at 0.15 m depth. Measurements were made of crop N uptake, residual fertilizer N and soil mineral N. The total above-ground dry matter yield of maize varied between 7.6 and 11.9 t ha^–1. The crop recovery of fertilizer N following point placement was 25% of that applied, which was higher than that from the surface application (18%) or incorporation by mixing (18%). The total grain yield of wheat varied between 4.3 and 4.7 t ha^–1. In the surface applications, the recovery of fertilizer-derived nitrogen (25%) was considerably lower than that from the mixing treatments and banded placements (33 and 36%). The fertilizer N application rate had a significant effect on grain and total dry matter yield, as well as on total N uptake and grain N contents. The main mechanism for loss of N appeared to be by ammonia volatilization, rather than leaching. High mineral N concentrations remained in the soil at harvest, following both crops, demonstrating a potential for significant reductions in N application rates without associated loss in yield.     

Statistical studies of branched ester latex and paint properties

Vinyl versatate, butyl acrylate and 2-ethylhexyl acrylate were compared as comonomer, for colloid-stabilized vinyl acetate latex polymers to assess their impact on latex coating properties. Vinyl versatate afforded advantages in scrub resistance, gloss, hydrophobicity and higher glass transition temperatures. Butyl acrylate developed better wet adhesion efficiency, gel content and hiding efficiency. 2-Ethylhexyl acrylate offered performance comparable to butyl acrylate in hiding efficiency, lowering copolymer glass transition temperatures, and gloss development. Combinations of these monomers, in many instances, afforded a better balance of performance properties than individual monomers alone. In the subject study, a simplex-centroid design was utilized to statistically map polymer compositions for determining the effect of comonomer composition on latex and paint film properties. In addition to developing contour maps of polymer and paint properties as a function of composition, a technique for studying structure-property relationships is presented.