NMR microscopy of single neurons at 500 MHZ

Previous NMR microimaging studies at 360 MHz have demonstrated a clear differentiation between the nucleus and cytoplasm in isolated single neurons. In particular, theT ₂ of the cell nucleus is 2.5 times larger than that of the cytoplasm. In order to determine the magnitude of possibleT ₂ ^* influences on these observations, images of single cells have been obtained at 500 MHz using spin-echo and line-narrowing sequences. Comparison of the images acquired by the two sequences, and of the spin-echo images at 360 and 500 MHz, imply that anyT ₂ ^* contributions are relatively small. Consequently, the measuredT ₂ differences in spin-echo imaging represent a true difference in theT ₂ relaxation in the two cellular compartments.     

Active screening in RF coil design

Active magnetic screening can be made to operate satisfactorily at RF frequencies up to approximately 8 MHz for small diameter short solenoids. The principles can be used to construct magnetically orthogonal coils which are geometrically coaxial. Such coils have a wide range of uses in NMR.     

Delignification of intact biomass and cellulosic coproduct of acid‐catalyzed hydrolysis

The kinetics of acid‐catalyzed hemicellulose removal and also alkaline delignification of oat hull biomass were investigated. All three operational parameters namely, catalyst concentration (0.10–0.55 N H₂SO₄), temperature (110–130°C), and residence time (up to 150 min) affected the efficiency of hemicellulose removal, with 100% of hemicellulose removed by appropriate selection of process parameters. Analysis of delignification kinetics (in the temperature range of 30–100°C) indicated that it can be expressed very well by a two‐phase model for the crude biomass and also for the hemicellulose‐prehydrolyzed material. The application of acid‐catalyzed prehydrolysis improved the capacity of lignin dissolution especially at lower temperatures (30 and 65°C) and accelerated the dissolution of lignin. This acceleration of delignification by prehydrolysis was possible at all levels of temperature in the bulk phase; however, results were more significant at the lower temperatures in the terminal phase. © 2015 American Institute of Chemical Engineers AIChE J, 61: 1783–1791, 2015     

Hydroxyoctadecadienoic Acids Regulate Apoptosis in Human THP‐1 Cells in a PPARγ‐Dependent Manner

Macrophage apoptosis, a key process in atherogenesis, is regulated by oxidation products, including hydroxyoctadecadienoic acids (HODEs). These stable oxidation products of linoleic acid (LA) are abundant in atherosclerotic plaque and activate PPARγ and GPR132. We investigated the mechanisms through which HODEs regulate apoptosis. The effect of HODEs on THP‐1 monocytes and adherent THP‐1 cells were compared with other C18 fatty acids, LA and α‐linolenic acid (ALA). The number of cells was reduced within 24 hours following treatment with 9‐HODE (p < 0.01, 30 μM) and 13 HODE (p < 0.01, 30 μM), and the equivalent cell viability was also decreased (p < 0.001). Both 9‐HODE and 13‐HODE (but not LA or ALA) markedly increased caspase‐3/7 activity (p < 0.001) in both monocytes and adherent THP‐1 cells, with 9‐HODE the more potent. In addition, 9‐HODE and 13‐HODE both increased Annexin‐V labelling of cells (p < 0.001). There was no effect of LA, ALA, or the PPARγ agonist rosiglitazone (1μM), but the effect of HODEs was replicated with apoptosis‐inducer camptothecin (10μM). Only 9‐HODE increased DNA fragmentation. The pro‐apoptotic effect of HODEs was blocked by the caspase inhibitor DEVD‐CHO. The PPARγ antagonist T0070907 further increased apoptosis, suggestive of the PPARγ‐regulated apoptotic effects induced by 9‐HODE. The use of siRNA for GPR132 showed no evidence that the effect of HODEs was mediated through this receptor. 9‐HODE and 13‐HODE are potent—and specific—regulators of apoptosis in THP‐1 cells. Their action is PPARγ‐dependent and independent of GPR132. Further studies to identify the signalling pathways through which HODEs increase apoptosis in macrophages may reveal novel therapeutic targets for atherosclerosis.     

Mechanistic Analysis of an Engineered Enzyme that Catalyzes the Formose Reaction

An enzyme that catalyzes the formose reaction, termed “formolase”, was recently engineered through a combination of computational protein design and directed evolution. We have investigated the kinetic role of the computationally designed residues and further characterized the enzyme's product profile. Kinetic studies illustrated that the computationally designed mutations were synergistic in their contributions towards enhancing activity. Mass spectrometry revealed that the engineered enzyme produces two products of the formose reaction—dihydroxyacetone and glycolaldehyde—with the product profile dependent on the formaldehyde concentration. We further explored the effects of this product profile on the thermodynamics and yield of the overall carbon assimilation from the formolase pathway to help guide future efforts to engineer this pathway.     

Effect of Oxime Ether Incorporation in Acyl Indole Derivatives on PPAR Subtype Selectivity

Compounds that simultaneously activate peroxisome proliferator‐activated receptor (PPAR) subtypes α and γ have the potential to effectively treat dyslipidemia and type 2 diabetes (T2D) in a single pharmaceutically active molecule. The frequently observed side effects of selective PPARγ agonists, such as edema and weight gain, were expected to be overcome by using additive PPARα activity, leading to dual PPARα/γ agonists with balanced activity for both subtypes. Herein we report the discovery, synthesis, and optimization of a new series of α‐ethoxyphenylpropionic acid bearing 5‐ or 6‐substituted indoles. The incorporation of oxime ethers on the carbonyl portion of the benzoyl group can bring the PPARα/γ potency ratio equal to or slightly greater than one, as is the case for compounds 20 c and 21 a . Compound 20 c shows high efficacy in an ob/ob mouse model of T2D and dyslipidemia, similar to that of rosiglitazone and tesaglitazar, but with a significant increase in body weight gain. In contrast, compound 21 a , less potent as a dual PPARα/γ activator than 20 c , showed an interesting pharmacological profile, as it elicits a decrease in body weight relative to reference compounds.     

Nitrogen fixation by pea and lentil green manures in a semi-arid agroecoregion: effect of planting and termination timing

Crop-fallow systems dominate many semi-arid agricultural regions despite fallow’s negative effects on soil and water quality. Annual legumes grown as a fallow-replacement crop, and terminated prior to maturity, can reduce these negative effects without substantially decreasing plant available water for the subsequent crop. Interest in growing legume green manures (LGMs) in synthetically-fertilized systems is increasing in the northern Great Plains of North America, partly due to the N-fixing capabilities of legumes; however, little is known about the effects of planting and termination time on N fixation amounts in the region. A 2-year field study was initiated in southwest Montana to determine the effects of planting time (spring or summer) and termination time (e.g. flower or pod) on the amount of N fixed by field pea (Pisum sativum cv. Arvika) and lentil (Lens culinaris cv. Richlea). Two methods, ¹⁵N natural abundance and N difference, were used to quantify N fixation, with wheat or in-crop weeds as reference plants. In 2009, N fixed by spring-planted lentil was higher by pod than flower (P = 0.03). Termination time did not affect the amount of N fixed by spring-planted pea, despite more biomass by pod than flower. In 2010, both spring-planted crops fixed more N by pod than flower (P < 0.01) and more N was fixed by spring-planted than summer-planted crops (P < 0.01). These results should prove useful to growers interested in selecting management practices that optimize N fixation of LGMs.     

Recovery and Functional Properties of Soy Storage Proteins from Lab- and Pilot-Plant Scale Oleosome Production

The aim of this study was to investigate soy protein recovery feasibility after lab- and pilot-plant scale oleosome isolation. The proteins were isolated by isoelectric precipitation and by ultrafiltration. The functional properties of the recovered proteins were compared to soy protein isolate produced in our laboratory. The residual lipid content in the aqueous supernatant affected the protein recovery yields and purities. Ethanol precipitation and ultrafiltration resulted in the best protein yields, which were 25 and 26% greater than protein yield obtained by isoelectric precipitation with distilled water dilution. The protein content of the isoelectric precipitated pilot-plant supernatant was higher (98%) than the protein content of ethanol-precipitated proteins (80%). Sodium dodecyl sulfate-polyacrylamide gel electrophoresis showed similar peptide profiles for laboratory and pilot-plant supernatants. Protein solubility curves between pH 3 and 8 were typical for soy protein isolate with higher solubilities for proteins obtained from pilot-plant supernatant. The soy protein isolate and ethanol-precipitated protein had the highest emulsification capacity on a dry-weight basis. These desirable functional properties of proteins recovered as co-products after oleosome isolation suggest they are highly suitable for industrial application as food ingredients and their recovery would contribute to the economics of the overall oleosome fractionation process.     

Additive-free hydrogelation of graphene oxide by ultrasonication

Graphene oxide hydrogels have been prepared by ultrasonication of precursor aqueous dispersions. The ultrasonication fractures the nanosheets, reducing their dimensions and exposing new sheet edges that do not possess the stabilizing carboxyl functional groups found along the edge of the as-prepared material. Ultrasonication does not affect the overall chemical functionality of the graphene oxide nanosheets, as spectra (carbon-13 nuclear magnetic resonance spectroscopy, infrared spectroscopy, and X-ray photoelectron spectroscopy) of samples before and after ultrasonication are nearly identical. Gelation is induced after only 30 min of ultrasonication to achieve a relatively weak gel with a shear modulus of 0.3 kPa; however, extension of ultrasonic treatment to 120 min yields a more robust hydrogel with a shear modulus of 1.6 kPa. Such enhancement in the gel’s physical properties can be attributed to the lack of stabilizing carboxyl groups on newly generated nanosheet fragments from the interior regions of the original nanosheets. As prepared, these hydrogels exhibit exceptionally low critical gelation concentrations ranging from ～0.050 to ～0.125 mg mL^?1 that can be tuned according to the extent of ultrasonic treatment.