Rapid Detection and Quantification of Triacylglycerol by HPLC–ELSD in Chlamydomonas reinhardtii and Chlorella Strains

Triacylglycerol (TAG) analysis and quantification are commonly performed by first obtaining a purified TAG fraction from a total neutral lipid extract using thin-layer chromatography (TLC), and then analyzing the fatty acid composition of the purified TAG fraction by gas chromatography (GC). This process is time-consuming, labor intensive and is not suitable for analysis of small sample sizes or large numbers. A rapid and efficient method for monitoring oil accumulation in algae using high performance liquid chromatography for separation of all lipid classes combined with detection by evaporative light scattering (HPLC–ELSD) was developed and compared to the conventional TLC/GC method. TAG accumulation in two Chlamydomonas reinhardtii (21 gr and CC503) and three Chlorella strains (UTEX 1230, CS01 and UTEX 2229) grown under conditions of nitrogen depletion was measured. The TAG levels were found to be 3–6 % DW (Chlamydomonas strains) and 7–12 % DW (Chlorella strains) respectively by both HPLC–ELSD and TLC/GC methods. HPLC–ELSD resolved the major lipid classes such as carotenoids, TAG, diacylglycerol (DAG), free fatty acids, phospholipids, and galactolipids in a 15-min run. Quantitation of TAG content was based on comparison to calibration curves of trihexadecanoin (16:0 TAG) and trioctadecadienoin (18:2 TAG) and showed linearity from 0.2 to 10 μg. Algal TAG levels >0.5 μg/g DW were detectable by this method. Furthermore TAG content in Chlorella kessleri UTEX 2229 could be detected. TAG as well as DAG and TAG content were estimated at 1.6 % DW by HPLC–ELSD, while it was undetectable by TLC/GC method.     

A cathode support structure for use in a magnetron oscillator experiment

A low temperature co-fired ceramic (LTCC) material system has been used to develop a protype field emission cathode structure for use in an experimental magnetron oscillator. The structure is designed for used with 30 gated field emission array (GFEA) die electrically connected through silver metal traces and electrical vias. To approximate a cylinder, the cathode structure (48 mm long and 13.7 mm in diameter) is comprised of 10 faceted plates which cover the GFEA dies. Slits in the facet plates allow electron injection. The GFEA die (3 mm × 8 mm) are placed in axial columns of 3 and spaced azimuthally around a cylindrical support structure in a staggered configuration resulting in 10 azimuthal locations. LTCC manufacturing techniques were developed in order to fabricate the newly designed cathode with seven layers wrapped to form the cylinder with electrical traces and vias. Two different cathode wrapping techniques and two different via filling techniques were studied and compared. Two different facet plate manufacturing techniques were studied. Finally, four different support stand configurations for firing the cylindrical structure were also compared with a square post stand having the best circularity and linearity measurements of the fired structure.     

Structural and functional analogy between pneumolysin and proaerolysin

Pneumolysin and proaerolysin are bacterial toxins that form pores in host cells by oligomerization. We propose that they may have similar structures despite a poor sequence identity. The crystal structure of proaerolysin reveals a protein composed of four domains, arranged in the shape of an elongated comma. Electron microscopy of the pneumolysin monomer shows a similar arrangement of domains. The sequence of pneumolysin recognizes the template of proaerolysin from a library of protein folds. A three-dimensional model of pneumolysin has been constructed by the comparative approach using the structure of proaerolysin. This model, together with results on the activity of site- specific mutants and the positions of antigenic sites, has been used to propose functional roles of individual domains.      

Production of human normal adult and fetal hemoglobins in Escherichia coli

A hemoglobin expression system in Escherichia coli is described. In order to produce authentic human hemoglobin, we need to co-express both methionine aminopeptidase and globin genes under the control of a strong promoter. We have constructed three plasmids, pHE2, pHE4 and pHE7, for the expression of human normal adult hemoglobin and a plasmid, pHE9, for the expression of human fetal hemoglobin, in high yields. The globin genes can be derived from either synthetic genes or human globin cDNAs. The extra amino-terminal methionine residues of the expressed globins can be removed by the co-expressed methionine aminopeptidase. The heme is inserted correctly into the expressed alpha- globin from our expression plasmids. A fraction (approximately 25%) of the heme is not inserted correctly into the expressed beta- or gamma- globin. However, the incorrectly inserted hemes can be converted into the correct conformation by carrying out a simple oxidation-reduction process on the purified hemoglobin molecule. We have investigated the functional properties of the expressed hemoglobins by measuring their oxygen-binding properties and their structural features by obtaining their 1H-NMR spectra. Our results show that authentic human normal adult and fetal hemoglobins can be produced from our expression plasmids in E. coli and in high yields. Our expression system allows us to design and to produce any recombinant hemoglobins needed for our research on the structure-function relationship in hemoglobin.      

Characterization of particle emissions from consumer fused deposition modeling 3D printers

Particle emissions from multiple fused deposition modeling consumer 3D printers were systematically quantified utilizing an established emission testing protocol (Blue Angel) to allow quantitative exposure assessments for printers operating in different environments. The data are consistent with particle generation from volatilization of the polymer filament as it is heated by the extruder. Typically, as printing begins, a burst of new particle formation leads to the smallest sizes and maximum number concentrations produced throughout the print job. For acrylonitrile butadiene styrene (ABS) filaments, instantaneous concentrations were up to 10⁶ #/cm³ with mean particle sizes of 20 to 40 nm when measured in a well mixed 1 m³ chamber with 1 air change per hour. Particles are continuously formed during printing and the size distribution evolves consistent with vapor condensation and particle coagulation. Particles emitted per mass of filament consumed (particle yield) varied widely due to factors including printer brand, and type and brand of filament. Higher extruder temperatures result in larger emissions. For filament materials tested, average particle number yields ranged from 7.3 × 10⁸ to 5.2 × 10¹⁰ g^?1 (approximately 0.65 to 24 ppm), with trace additives apparently driving the large variations. Nanoparticles (diameters less than 100 nm) dominate number distributions, whereas diameters in the range of 200 to 500 nm contribute most to estimated mass. Because 3D printers are often used in public spaces and personal residences, the general public and particularly susceptible populations, such as children, can be exposed to high concentrations of non-engineered nanoparticles of potential toxicity.

Copyright © 2017 American Association for Aerosol Research     

Dislocations at Prismatic Fracture Surfaces in Sapphire

Single-crystal sapphire exhibits a highly anisotropic fracture behavior. The surfaces of specimens fractured along prismatic planes are wavy, with fractographic features appearing as small areas of contrast under an optical microscope. Optical microscopy, atomic force microscopy, X-ray topography, and confocal microscopy are used to demonstrate a correlation between the areas of contrast and dislocations penetrating the sample surface. The surface features are argued to be a consequence of the stress field surrounding the dislocations, which deflect the crack approximately 10 nm normal to the surface as the crack cuts the dislocations. The lateral extent of measurable surface deflection is of the order of 5 μm. These images can be compared directly to show the equivalence of the position of the dislocations observed by X-ray topography and the surface contrast observed optically.     

Metallocene QACs: The Incorporation of Ferrocene Moieties into monoQAC and bisQAC Structures

Inspired by the incorporation of metallocene functionalities into a variety of bioactive structures, particularly antimicrobial peptides, we endeavored to broaden the structural variety of quaternary ammonium compounds (QACs) by the incorporation of the ferrocene moiety. Accordingly, 23 ferrocene-containing mono- and bisQACs were prepared in high yields and tested for activity against a variety of bacteria, including Gram-negative strains and a panel of clinically isolated MRSA strains. Ferrocene QACs were shown to be effective antiseptics with some displaying single-digit micromolar activity against all bacteria tested, demonstrating yet another step in the expansion of structural variety of antiseptic QACs.     

LIQUID PHASE CO-OXIDATION OF THIOPHENOL AND STYRENE BY OXYGEN AND t-BUTYL HYDROPEROXIDE

ABSTRACT

Instability problems in both shale and petroleum derived middle distillate jet fuels have been correlated with the presence of peroxidic species. Although a good body of knowledge exists concerning the formation of peroxides in the liquid phase, relatively little is known about the reaction/ decomposition pathways available when other functional groups are present, since sulfur is the most abundant heteroatam present in jet fuels, the reaction of t-butyl hydroperoxide (tBKP) and/or oxygen with thiophenol in the presence of the active olefin, styrene, was examined in deaerated benzene at 120°C. The complex product mixture was analyzed by combined capillary column GC/MS. Major products included acetone, t-butanol and isdbutylene from the tBHP. Thiophenol and styrene combined to form addition products. Phenyl disulfide was observed from the thiophenol. The results indicated that although the product slate was complex, it was possible to explain the product mix in terms of a few competing reactions.     

Particle shape control using pulse electrodeposition: Methanol oxidation as a probe reaction on Pt dendrites and cubes

Shape controlled Pt particles were synthesized onto tungsten monocarbide (WC) substrates using a pulse electrodeposition method. The particle shape was strongly influenced by the deposition potential, with cubic particles formed using 0.14 V vs. NHE (normal hydrogen electrode) and dendritic particles formed at 0.04 V vs. NHE. The crystalline orientation and active surface area of the Pt particles were estimated using Cu stripping voltammetry. Finally, cyclic voltammetry and chronoamperometry were used to determine the methanol electrooxidation activity, which revealed that the dendritic Pt showed much higher electrochemical activity than the cubic particles. These results demonstrated the possibility of more effectively utilizing Pt electrocatalysts by controlling the shape of Pt particles.     

EFFECT OF A MAJOR FLOOD ON BREEDING AND HABITAT OF THE CRIMSON FINCH (NEOCHMIA PHAETON): A RIPARIAN SPECIALIST

Riparian specialists, such as the Crimson Finch (Estrildidae: Neochmia phaeton), are vulnerable to declines in habitat quality, including alterations of flow regime associated with dam construction. Crimson Finches persisted and bred in substantial numbers following two large floods in the Isaac–Connors catchment in early 2008. Major flooding was not detrimental to nesting and breeding success, although the immediate post‐flood period was identified as a bottleneck in the availability of riparian grass seed. Crimson Finches nested at 16 m or more above the normal river level (compared with an average of 2–3 m elsewhere in Northern Australia); indicating that they have adapted to the extreme flood peaks in the Fitzroy River basin. The tall river‐associated grass Chionachne cyathopoda was a key habitat plant, retaining seed well into the dry season when most other grasses are expended. It also provided abundant seeds following floods, coinciding with peaks in abundance of dependent young finches. The adaptations of both Crimson Finches and Chionachne suggest that increased flooding, predicted by climate change, is unlikely to be a problem. In contrast, a proposed dam on the Connors River may be a threat. River regulation that reduces peaks of major floods may favour introduced riparian grasses over flood adapted native grasses such as Chionachne. Any change in the quantity of this species could be detrimental to persistence of Crimson Finches. Environmental managers should carefully consider the ecology of Chionachne when making decisions about river flows. Copyright © 2013 John Wiley & Sons, Ltd.