Metabolomics reveals increased isoleukotoxin diol (12,13-DHOME) in human plasma after acute Intralipid infusion

Intralipid is a fat emulsion that is regularly infused into humans and animals. Despite its routine use, Intralipid infusion can cause serious adverse reactions, including immunosuppression. Intralipid is a complex mix of proteins, lipids, and other small molecules, and the effect of its infusion on the human plasma metabolome is unknown. We hypothesized that untargeted metabolomics of human plasma after an Intralipid infusion would reveal novel insights into its effects. We infused Intralipid and saline into 10 healthy men in a double-blind, placebo-controlled experiment and used GC/MS, LC/MS, and NMR to profile the small-molecule composition of their plasma before and after infusion. Multivariate statistical analysis of the 40 resulting plasma samples revealed that after Intralipid infusion, a less-well-characterized pathway of linoleic acid metabolism had resulted in the appearance of (9Z)-12,13-dihydroxyoctadec-9-enoic acid (12,13-DHOME, P < 10(-3)), a leukotoxin that has powerful physiological effects and is known to inhibit the neutrophil respiratory burst. Intralipid infusion caused increased plasma 12,13-DHOME. Given that 12,13-DHOME is known to directly affect neutrophil function, we conclude that untargeted metabolomics may have revealed a hitherto-unknown mechanism of intralipid-induced immunosuppression.     

First record of preserved soft parts in a Palaeozoic podocopid (Metacopina) ostracod, Cytherellina submagna: phylogenetic implications

The metacopines represent one of the oldest and most important extinct groups of ostracods, with a fossil record from the Mid-Ordovician to the Early Jurassic. Herein, we report the discovery of a representative of the group with three-dimensionally preserved soft parts. The specimen--a male of Cytherellina submagna--was found in the Early Devonian (416 Ma) of Podolia, Ukraine. A branchial plate (Bp) of the cephalic maxillula (Mx), a pair of thoracic appendages (walking legs), a presumed furca (Fu) and a copulatory organ are preserved. The material also includes phosphatized steinkerns with exceptionally preserved marginal pore canals and muscle scars. The morphology of the preserved limbs and valves of C. submagna suggests its relationship with extant Podocopida, particularly with the superfamilies Darwinuloidea and Sigillioidea, which have many similar characteristic features, including a large Bp on the Mx, the morphology of walking legs, Fu with two terminal claws, internal stop-teeth in the left valve, adductor muscle scar pattern, and a very narrow fused zone along the anterior and posterior margins. More precise determination of affinities will depend on the soft-part morphology of the cephalic segment, which has not been revealed in the present material.     

Computational modeling-based discovery of novel classes of anti-inflammatory drugs that target lanthionine synthetase C-like protein 2

Background

Lanthionine synthetase component C-like protein 2 (LANCL2) is a member of the eukaryotic lanthionine synthetase component C-Like protein family involved in signal transduction and insulin sensitization. Recently, LANCL2 is a target for the binding and signaling of abscisic acid (ABA), a plant hormone with anti-diabetic and anti-inflammatory effects.

Methodology/Principal Findings

The goal of this study was to determine the role of LANCL2 as a potential therapeutic target for developing novel drugs and nutraceuticals against inflammatory diseases. Previously, we performed homology modeling to construct a three-dimensional structure of LANCL2 using the crystal structure of lanthionine synthetase component C-like protein 1 (LANCL1) as a template. Using this model, structure-based virtual screening was performed using compounds from NCI (National Cancer Institute) Diversity Set II, ChemBridge, ZINC natural products, and FDA-approved drugs databases. Several potential ligands were identified using molecular docking. In order to validate the anti-inflammatory efficacy of the top ranked compound (NSC61610) in the NCI Diversity Set II, a series of in vitro and pre-clinical efficacy studies were performed using a mouse model of dextran sodium sulfate (DSS)-induced colitis. Our findings showed that the lead compound, NSC61610, activated peroxisome proliferator-activated receptor gamma in a LANCL2- and adenylate cyclase/cAMP dependent manner in vitro and ameliorated experimental colitis by down-modulating colonic inflammatory gene expression and favoring regulatory T cell responses.

Conclusions/Significance

LANCL2 is a novel therapeutic target for inflammatory diseases. High-throughput, structure-based virtual screening is an effective computational-based drug design method for discovering anti-inflammatory LANCL2-based drug candidates.     

Bitter taste of saccharin and acesulfame-K.

The relationships among suprathreshold taste responses to acesulfame-K, Na-saccharin and 6-n-propylthiouracil (PROP) were examined in two studies. In the first study, the labeled magnitude scale was used with the high anchor labeled as 'strongest imaginable oral sensation' and in the second study, it was labeled as 'strongest imaginable sensation of any kind'. Results from the two procedures were similar. Individual differences among 65 subjects were seen in bitter responses to acesulfame-K and saccharin. Bitter responses to acesulfame-K ands accharin were positively correlated, but showed no significant relationship with responses to PROP bitterness or with PROP taster groups. Saccharin and acesulfame-K may share a common mechanism for bitter taste reception and transduction, one that varies across individuals and is different from mechanisms mediating bitter responses to PROP. Changing the instructions of the labeled magnitude scale induced a context effect. Ratings of sweetness referenced to the 'strongest imaginable sensationof any kind' were lower than ratings referenced to just oral sensations.     

Comparison of the biochemical and kinetic properties of the type 1 receptor tyrosine kinase intracellular domains. Demonstration of differential sensitivity to kinase inhibitors.

Epidermal growth factor receptor (EGFR), ErbB-2, and ErbB-4 are members of the type 1 receptor tyrosine kinase family. Overexpression of these receptors, especially ErbB-2 and EGFR, has been implicated in multiple forms of cancer. Inhibitors of EGFR tyrosine kinase activity are being evaluated clinically for cancer therapy. The potency and selectivity of these inhibitors may affect the efficacy and toxicity of therapy. Here we describe the expression, purification, and biochemical comparison of EGFR, ErbB-2, and ErbB-4 intracellular domains. Despite their high degree of sequence homology, the three enzymes have significantly different catalytic properties and substrate kinetics. For example, the catalytic activity of ErbB-2 is less stable than that of EGFR. ErbB-2 uses ATP-Mg as a substrate inefficiently compared with EGFR and ErbB-4. The three enzymes have very similar substrate preferences for three optimized peptide substrates, but differences in substrate synergies were observed. We have used the biochemical and kinetic parameters determined from these studies to develop an assay system that accurately measures inhibitor potency and selectivity between the type 1 receptor family. We report that the selectivity profile of molecules in the 4-anilinoquinazoline series can be modified through specific aniline substitutions. Moreover, these compounds have activity in whole cells that reflect the potency and selectivity of target inhibition determined with this assay system.     

Prediction of protein helix content from an autocorrelation analysis of sequence hydrophobicities

It is demonstrated that protein α-helix content can be predicted from an autocorrelation analysis of the protein hydrophobicity sequence. The Fourier transform of the autocorrelation function yields the spectral densities or weights of the various frequencies contributing to the autocorrelation function. Using sequence and secondary structure data from more than 160 proteins and domains, a linear relationship was found between spectral density at periodicity 3.7 and protein α-helix content (r = 0.83). This relation permits prediction of the helix content (x) of proteins of known sequence to within ± 15%, i.e., as (x ± 15)%. Predictions based on the autocorrelation procedure are compared with values obtained by other methods.     

Kinetics of aggregation of αs1 -casein/ca2+ mixtures: charge and temperature effects

Time-dependent light-scattering studies have been made on mixtures of α_s1 -casein and Ca²⁺ at fixed temperature over a range of [Ca²⁺] and [α_s1 -casein], and also as functions of temperature- Measurements were also made of the extent of precipitate formation in the casein/Ca²⁺ mixtures, using centrifugation. The results are analysed in terms of a monomeroctamer equilibrium between calcium caseinate particles followed by a Smoluchowski aggregation in which only the octamers can participate. The equilibrium constant is dependent upon the charge on the protein/Ca²⁺ particles, and hence can be related to the extent of binding of Ca²⁺ to the α_s1 -casein. The Smoluchowski constant is likewise shown to be charge-dependent. The variation of the reaction rate with temperature can be ascribed solely to the changing charge of the α_s1 -casein/Ca²⁺ complex caused by changed binding of Ca²⁺ at different temperatures.