Neutron-capture element deficiency of the Hercules dwarf spheroidal galaxy

We present an assessment of the barium abundance ratios for red giant member stars in the faint Hercules dwarf spheroidal (dSph) galaxy. Our results are drawn from intermediate-resolution FLAMES/GIRAFFE spectra around the Ba II 6141.71 AA absorption line at low signal-to-noise ratios. For three brighter stars we were able to gain estimates from direct equivalent-width measurements, while for the remaining eight stars only upper limits could be obtained. These results are investigated in a statistical manner and indicate very low Ba abundances of log epsilon (Ba) < 0.7 dex (3 sigma). We discuss various possible systematic biasses, first and foremost, a blend with the Fe I 6141.73 AA-line, but most of those would only lead to even lower abundances. A better match with metal-poor halo and dSph stars can only be reached by including a large uncertainty in the continuum placement. This contrasts with the high dispersions in iron and calcium (in excess of 1 dex) in this galaxy. While the latter spreads are typical of the very low luminosity, dark-matter dominated dSphs, a high level of depletion in heavy elements suggests that chemical enrichment in Hercules was governed by very massive stars, coupled with a very low star formation efficiency. While very low abundances of some heavy elements are also found in individual stars of other dwarf galaxies, this is the first time that a very low Ba abundance is found within an entire dSph over a broad metallicity range.Comment: 10 pages, 6 figures, accepted for publication in Astronomy & Astrophysic

Effect of Wakame and Carob Pod Snacks on Non-Alcoholic Fatty Liver Disease

Snacks combining different functional ingredients could represent a useful therapeutic strategy against NAFLD. The present study aimed to analyze the effect of two snack formulations based on carob and wakame flour in the treatment for NAFLD in rats. For this purpose, metabolic syndrome was induced in 50 adult rats by a high-fat high-fructose diet over eight weeks. After this period, rats were fed either normal calorie diets supplemented or not with snack A (1/50 wakame/carob pod) and snack B (1/5 wakame/carob pod) for four additional weeks. After sacrifice, liver composition and serum parameters were analyzed. Different pathways of triacylglycerol metabolism in liver were studied including fatty acid oxidation, fatty acid synthesis, triglyceride assembly and release, fatty acid uptake and glucose uptake. Oxidative stress was also measured. Snack treatment, and mainly B snack, reduced liver triacylglycerol levels by increasing fat oxidation. Moreover, this snack reduced oxidative stress. Therefore, this snack formulation could represent an interesting tool useful for fatty liver treatment.This study has been supported by the National Institute for Agricultural and Food Research and Technology of Spain (INIA: RTA2014-0037-C02), Instituto de Salud Carlos III (CIBERobn) and Basque Government (IT-572-13)

Transient Expression of Tetrameric Recombinant Human Butyrylcholinesterase in Nicotiana benthamiana.

To optimize the expression, extraction and purification of plant-derived tetrameric recombinant human butyrylcholinesterase (prBChE), we describe the development and use of plant viral amplicon-based gene expression system; Tobacco Mosaic Virus (TMV) RNA-based overexpression vector (TRBO) to express enzymatically active FLAG-tagged plant made recombinant butyrylcholinesterase (rBChE) in Nicotiana benthamiana leaves using transient agroinfiltration. Two gene expression cassettes were designed to express the recombinant protein in either the ER or to the apoplastic compartment. Leaf homogenization was used to isolate ER-retained recombinant butyrylcholinesterase (prBChE-ER) while apoplast-targeted rBChE was isolated by either leaf homogenization (prBChE) or vacuum-extraction of apoplastic wash fluid (prBChE-AWF). rBChE from apoplast wash fluid had a higher specific activity but lower enzyme yield than leaf homogenate. To optimize the isolation and purification of total recombinant protein from leaf homogenates, an acidic extraction buffer was used. The acidic extraction buffer yielded &gt;95% enzymatically active tetrameric rBChE as verified by Coomassie stained and native gel electrophoresis. Furthermore, when compared to human butyrylcholinesterase, the prBChE was found to be similar in terms of tetramerization and enzyme kinetics. The N-linked glycan profile of purified prBChE-ER was found to be mostly high mannose structures while the N-linked glycans on prBChE-AWF were primarily complex. The glycan profile of the prBChE leaf homogenates showed a mixture of high mannose, complex and paucimannose type N-glycans. These findings demonstrate the ability of plants to produce rBChE that is enzymatically active and whose oligomeric state is comparable to mammalian butyrylcholinesterase. The process of plant made rBChE tetramerization and strategies for improving its pharmacokinetics properties are also discussed

Leucine supplementation differentially enhances pancreatic cancer growth in lean and overweight mice

Kristyn A Liu1†, Laura M Lashinger1†, Audrey J Rasmussen1† and Stephen D Hursting12* Author Affiliations 1 Department of Nutritional Sciences, University of Texas at Austin, Austin, TX 78723, USA 2 Department of Molecular Carcinogenesis, University of Texas M.D. Anderson Cancer Center, 1808 Park Road 1c, Smithville, TX 78957, USABackground: The risk of pancreatic cancer, the 4th deadliest cancer for both men and women in the United States, is increased by obesity. Calorie restriction (CR) is a well-known dietary regimen that prevents or reverses obesity and suppresses tumorigenesis in a variety of animal models, at least in part via inhibition of mammalian target of rapamycin (mTOR) signaling. Branched-chain amino acids (BCAA), especially leucine, activate mTOR and enhance growth and proliferation of myocytes and epithelial cells, which is why leucine is a popular supplement among athletes. Leucine is also increasingly being used as a treatment for pancreatic cancer cachexia, but the effects of leucine supplementation on pancreatic tumor growth have not been elucidated. Results: Supplementation with leucine increased pancreatic tumor growth in both lean (104 ± 17 mm3 versus 46 ± 13 mm3; P <0.05) and overweight (367 ± 45 mm3 versus 230 ± 39 mm3; P <0.01) mice, but tumor enhancement was associated with different biological outcomes depending on the diet. In the lean mice, leucine increased phosphorylation of mTOR and downstream effector S6 ribosomal protein, but in the overweight mice, leucine reduced glucose clearance and thus increased the amount of circulating glucose available to the tumor. Conclusion: These findings show that leucine supplementation enhances tumor growth in both lean and overweight mice through diet-dependent effects in a murine model of pancreatic cancer, suggesting caution against the clinical use of leucine supplementation for the purposes of skeletal muscle enhancement in cachectic patients.Nutritional Science

Cloning, Expression, Sequence Analysis and Homology Modeling of the Prolyl Endoprotease from Eurygaster integriceps Puton

Eurygaster integriceps Puton, commonly known as sunn pest, is a major pest of wheat in Northern Africa, the Middle East and Eastern Europe. This insect injects a prolyl endoprotease into the wheat, destroying the gluten. The purpose of this study was to clone the full length cDNA of the sunn pest prolyl endoprotease (spPEP) for expression in E. coli and to compare the amino acid sequence of the enzyme to other known PEPs in both phylogeny and potential tertiary structure. Sequence analysis shows that the 5ꞌ UTR contains several putative transcription factor binding sites for transcription factors known to be expressed in Drosophila that might be useful targets for inhibition of the enzyme. The spPEP was first identified as a prolyl endoprotease by Darkoh et al., 2010. The enzyme is a unique serine protease of the S9A family by way of its substrate recognition of the gluten proteins, which are greater than 30 kD in size. At 51% maximum identity to known PEPs, homology modeling using SWISS-MODEL, the porcine brain PEP (PDB: 2XWD) was selected in the database of known PEP structures, resulting in a predicted tertiary structure 99% identical to the porcine brain PEP structure. A Km for the recombinant spPEP was determined to be 210 ± 53 µM for the zGly-Pro-pNA substrate in 0.025 M ethanolamine, pH 8.5, containing 0.1 M NaCl at 37 °C with a turnover rate of 172 ± 47 µM Gly-Pro-pNA/s/µM of enzyme

Acetylene terminated matrix resins

The synthesis of resins with terminal acetylene groups has provided a promising technology to yield high performance structural materials. Because these resins cure through an addition reaction, no volatile by-products are produced during the processing. The cured products have high thermal stability and good properties retention after exposure to humidity. Resins with a wide variety of different chemical structures between the terminal acetylene groups are synthesized and their mechanical properties studied. The ability of the acetylene cured polymers to give good mechanical properties is demonstrated by the resins with quinoxaline structures. Processibility of these resins can be manipulated by varying the chain length between the acetylene groups or by blending in different amounts of reactive deluents. Processing conditions similar to the state-of-the-art epoxy can be attained by using backbone structures like ether-sulfone or bis-phenol-A. The wide range of mechanical properties and processing conditions attainable by this class of resins should allow them to be used in a wide variety of applications

Complete element abundances of nine stars in the r-process galaxy Reticulum II

We present chemical abundances derived from high-resolution Magellan/MIKE spectra of the nine brightest known red giant members of the ultra-faint dwarf galaxy Reticulum II. These stars span the full metallicity range of Ret II (-3.5 < [Fe/H] < -2). Seven of the nine stars have extremely high levels of r-process material ([Eu/Fe]~1.7), in contrast to the extremely low neutron-capture element abundances found in every other ultra-faint dwarf galaxy studied to date. The other two stars are the most metal-poor stars in the system ([Fe/H] < -3), and they have neutron-capture element abundance limits similar to those in other ultra-faint dwarf galaxies. We confirm that the relative abundances of Sr, Y, and Zr in these stars are similar to those found in r-process halo stars but ~0.5 dex lower than the solar r-process pattern. If the universal r-process pattern extends to those elements, the stars in Ret II display the least contaminated known r-process pattern. The abundances of lighter elements up to the iron peak are otherwise similar to abundances of stars in the halo and in other ultra-faint dwarf galaxies. However, the scatter in abundance ratios is large enough to suggest that inhomogeneous metal mixing is required to explain the chemical evolution of this galaxy. The presence of low amounts of neutron-capture elements in other ultra-faint dwarf galaxies may imply the existence of additional r-process sites besides the source of r-process elements in Ret II. Galaxies like Ret II may be the original birth sites of r-process enhanced stars now found in the halo.Comment: 14 pages, 5 figures, 5 tables. Accepted to Ap

Grifonin-1: A Small HIV-1 Entry Inhibitor Derived from the Algal Lectin, Griffithsin

Background: Griffithsin, a 121-residue protein isolated from a red algal Griffithsia sp., binds high mannose N-linked glycans of virus surface glycoproteins with extremely high affinity, a property that allows it to prevent the entry of primary isolates and laboratory strains of T- and M-tropic HIV-1. We used the sequence of a portion of griffithsin's sequence as a design template to create smaller peptides with antiviral and carbohydrate-binding properties. Methodology/Results: The new peptides derived from a trio of homologous β-sheet repeats that comprise the motifs responsible for its biological activity. Our most active antiviral peptide, grifonin-1 (GRFN-1), had an EC50 of 190.8±11.0 nM in in vitro TZM-bl assays and an EC50 of 546.6±66.1 nM in p24gag antigen release assays. GRFN-1 showed considerable structural plasticity, assuming different conformations in solvents that differed in polarity and hydrophobicity. Higher concentrations of GRFN-1 formed oligomers, based on intermolecular β-sheet interactions. Like its parent protein, GRFN-1 bound viral glycoproteins gp41 and gp120 via the N-linked glycans on their surface. Conclusion: Its substantial antiviral activity and low toxicity in vitro suggest that GRFN-1 and/or its derivatives may have therapeutic potential as topical and/or systemic agents directed against HIV-1