Discovery of Ubonodin,an Antimicrobial Lasso Peptide Active against Members of the Burkholderia cepacia Complex

We report the heterologous expression, structure, and antimicrobial activity of a lasso peptide, ubonodin, encoded in the genome of Burkholderia ubonensis. The topology of ubonodin is unprecedented amongst lasso peptides, with 18 of its 28 amino acids found in the mechanically bonded loop segment. Ubonodin inhibits RNA polymerase in vitro and has potent antimicrobial activity against several pathogenic members of the Burkholderia genus, most notably B. cepacia and B. multivorans, causative agents of lung infections in cystic fibrosis patients.     

Propellant cohesive fracture during the peel test of a propellant/liner structure

The integrity of propellant/liner structures in rocket motors is critical to ensure controlled combustion of the engine. In an effort to improve the bonding between the liner and the propellant, it is necessary to characterize it well. Therefore, a propellant–liner structure, bounded thanks to co-curing, has been submitted to a peel test while recording the macroscopic fracture energy and the local displacement field on the propellant-free surface. The experimental setup includes two cameras in order to record the displacement field on the propellant-free surface. Upon loading, the peel force stabilizes quickly due to a cohesive fracture in the propellant, providing access to the fracture energy. While the crack propagates through the propellant, it is observed that only a small localized area is submitted to strain, and most of the structure remains unstrained.     

Validation of a Method for Quantitation of Soybean Lectin in Commercial Varieties

Soybean agglutinin (SBA) protein, also known as soybean lectin, is regarded as an anti‐nutrient due to its negative effect on the ability of monogastric animals to gain weight following consumption of raw soybean seed. Historically, SBA has been measured using a time‐consuming and cumbersome hemagglutination procedure. The objective of our research was to obtain a validated methodology that is precise and accurate in the measurement of SBA while allowing minimally equipped laboratories to effectively conduct the analysis, thus our focus was on evaluating an existing commercially available ELISA, an enzyme‐linked‐lectin‐assay (ELLA), and a hybrid ELISA/ELLA. A new ELLA technique that can detect and quantify lectins was chosen and modified specifically for the quantitation of SBA in soybean seed. The proposed ELLA methodology is similar to a standard sandwich ELISA, and uses polyacrylamide‐linked N‐acetylgalactosamine (Gal–NAc–PAA) for a capture phase and the biotinylated version (Gal–NAc–PAA–Biotin) for detection. Based upon the validation data, the ELLA method can precisely and accurately determine soybean lectin levels in soybean seed. The validated ELLA method was used to quantify SBA in nine commercial soybean varieties introduced between 1972 and 2008 and demonstrated that the natural variability of SBA is subject to the effects of genotype and environment.     

Tetrahydroisoquinolines: New Inhibitors of Neutrophil Extracellular Trap (NET) Formation

Neutrophils are short‐lived leukocytes that migrate to sites of infection as part of the acute immune response, where they phagocytose, degranulate, and form neutrophil extracellular traps (NETs). During NET formation, the nuclear lobules of neutrophils disappear and the chromatin expands and, accessorized with neutrophilic granule proteins, is expelled. NETs can be pathogenic in, for example, sepsis, cancer, and autoimmune and cardiovascular diseases. Therefore, the identification of inhibitors of NET formation is of great interest. Screening of a focused library of natural‐product‐inspired compounds by using a previously validated phenotypic NET assay identified a group of tetrahydroisoquinolines as new NET formation inhibitors. This compound class opens up new avenues for the study of cellular death through NET formation (NETosis) at different stages, and might inspire new medicinal chemistry programs aimed at NET‐dependent diseases.     

A Blend of Ethanol and (−)-α-Pinene were Highly Attractive to Native Siricid Woodwasps (Siricidae,Siricinae) Infesting Conifers of the Sierra Nevada and the Allegheny Mountains

Woodwasps in Sirex and related genera are well-represented in North American conifer forests, but the chemical ecology of native woodwasps is limited to a few studies demonstrating their attraction to volatile host tree compounds, primarily monoterpene hydrocarbons and monoterpene alcohols. Thus, we systematically investigated woodwasp-host chemical interactions in California’s Sierra Nevada and West Virginia’s Allegheny Mountains. We first tested common conifer monoterpene hydrocarbons and found that (?)-α-pinene, (+)-3-carene, and (?)-β-pinene were the three most attractive compounds. Based on these results and those of earlier studies, we further tested three monoterpene hydrocarbons and four monoterpene alcohols along with ethanol in California: monoterpene hydrocarbons caught 72.3% of all woodwasps. Among monoterpene hydrocarbons, (+)-3-carene was the most attractive followed by (?)-β-pinene and (?)-α-pinene. Among alcohols, ethanol was the most attractive, catching 41.4% of woodwasps trapped. Subsequent tests were done with fewer selected compounds, including ethanol, 3-carene, and ethanol plus (?)-α-pinene in both Sierra Nevada and Allegheny Mountains. In both locations, ethanol plus (?)-α-pinene caught more woodwasps than other treatments. We discussed the implications of these results for understanding the chemical ecology of native woodwasps and invasive Sirex noctilio in North America. In California, 749 woodwasps were caught, representing five species: Sirex areolatus Cresson, Sirex behrensii Cresson, Sirex cyaneus Fabricius, Sirex longicauda Middlekauff, and Urocerus californicus Norton. In West Virginia 411 woodwasps were caught representing four species: Sirex edwardsii Brullé, Tremex columba Linnaeus, Sirex nigricornis F., and Urocerus cressoni Norton.     

Combined Extraction of Cesium and Strontium from Alkaline Nitrate Solutions

Abstract

The combined extraction of cesium and strontium from caustic wastes can be achieved by adding a crown ether and a carboxylic acid to the Caustic‐Side Solvent Extraction (CSSX) solvent. The ligand 4,4′(5′)‐di(tert‐butyl)cyclohexano‐18‐crown‐6 and one of four different carboxylic acids were combined with the components of the CSSX solvent optimized for the extraction of cesium, allowing for the simultaneous extraction of cesium and strontium from alkaline nitrate media simulating alkaline high level wastes present at the U.S. Department of Energy Savannah River Site. Extraction and stripping experiments were conducted independently and exhibited adequate results for mimicking waste simulant processing through batch contacts. The promising results of these batch tests showed that the system could reasonably be tested on actual waste.     

Stress Analysis of Adhesively Bonded Electroprimed Steel Lap Shear Joints

Structural applications for adhesive bonding have been increasing in recent years due to improvements in the types of adhesives available and in improved knowledge of bonding procedures. Consequently, there exists a demand for precise numerical modeling of adhesive joint behavior, particularly along bondline interfaces where low surface energy adhesives contact high surface energy metallic oxides. The purpose of the present study is to determine the effect of electrodeposited organic paint primer (ELPO) on the stress and strain distributions within an adhesively bonded single-lap-shear joint. Initial experimental studies have shown that bonding to ELPO-primed steel adherends has enhanced strength and durability characteristics compared to conventional bonds to unprimed steel surfaces. Recent studies based on finite element analysis of varied single-lap-shear joint moduli and thicknesses, and subsequent testing of joints with two different adhesive moduli, have indicated the mechanisms involved in this phenomenon. The presence of the ELPO-primer reduced peak peel and shear stresses and allowed for more uniform stress distribution throughout the joint.     

Comparison of the Chemical and Oxidative Characteristics of Particulate Matter (PM) Collected by Different Methods: Filters,Impactors, and BioSamplers

In this study, we compare the chemical and oxidative characteristics of concentration-enriched PM_2.5 samples simultaneously collected by a filter, a Nano-Micro-Orifice Uniform Deposition Impactor, and a BioSampler. Gravimetric measurements showed considerable agreement in particulate matter (PM) collection efficiency for all three samplers. Accordingly, samples from the three collectors exhibited similar chemical compositions. The mass fractions of their inorganic ions, labile and nonlabile, were comparable. Moreover, the organic carbon (OC) content of the BioSampler slurry was similar to that of the filter, while water-soluble OC levels of the filter and impactor samples were close to a 100% agreement. Lastly, linear regression analyses demonstrated that the water-soluble elements existed in similar proportions for the filter and impactor samples. Their respective total components were also in very good agreement. By contrast, the recoverable elements from the BioSampler slurry, determined by high-resolution magnetic sector inductively coupled plasma mass spectrometry, were in good agreement with the water-soluble elements of the filter and impactor samples but not their corresponding total components. In spite of the overall agreement among the samples on their chemical composition, findings from a macrophage reactive oxygen species (ROS), a dithiothreitol (DTT), and a dihydroxybenzoate (DHBA) assay revealed that the oxidative potential of aqueous extracts of the filter and impactor substrates was similar yet substantially lower than that of the BioSampler slurry. However, filtering of the BioSampler slurry, i.e., removal of insoluble PM components, attenuated its ROS activity to about the same level as that of the water extracts of the filter and impactor samples. These findings first indicate that insoluble PM species are potentially redox active, and second that particle collection by the BioSampler, which circumvents the need for PM extraction, constitutes a viable alternative for collecting concentrated particles for characterization of the oxidative properties of PM.