Prediction of a New Ligand-Binding Site for Type 2 Motif based on the Crystal Structure of ALG-2 by Dry and Wet Approaches

ALG-2 is a penta-EF-hand Ca(2+)-binding protein and interacts with a variety of intracellular proteins. Two types of ALG-2-binding motifs have been determined: type 1, PXYPXnYP (X, variable; n = 4), in ALIX and PLSCR3; type 2, PXPGF, in Sec31A and PLSCR3. The previously solved X-ray crystal structure of the complex between ALG-2 and an ALIX peptide containing type 1 motif showed that the peptide binds to Pocket 1 and Pocket 2. Co-crystallization of ALG-2 and type 2 motif-containing peptides has not been successful. To gain insights into the molecular basis of type 2 motif recognition, we searched for a new hydrophobic cavity by computational algorithms using MetaPocket 2.0 based on 3D structures of ALG-2. The predicted hydrophobic pocket designated Pocket 3 fits with N-acetyl-ProAlaProGlyPhe-amide, a virtual penta-peptide derived from one of the two types of ALG-2-binding sites in PLSCR3 (type 2 motif), using the molecular docking software AutoDock Vina. We investigated effects of amino acid substitutions of the predicted binding sites on binding abilities by pulldown assays using glutathione-S-transferase -fused ALG-2 of wild-type and mutant proteins and lysates of cells expressing green fluorescent protein -fused PLSCR3 of wild-type and mutants. Substitution of either L52 with Ala or F148 with Ser of ALG-2 caused loss of binding abilities to PLSCR3 lacking type 1 motif but retained those to PLSCR3 lacking type 2 motif, strongly supporting the hypothesis that Pocket 3 is the binding site for type 2 motif.     

Novel Heneicosadienoic and Tricosadienoic Acid Isomers in Ovaries of Marine Archaeogastropods

To investigate unusual odd-chain fatty acids (FA) from gonads of archaeogastropods, limpets Cellana grata and Cellana toreuma, a subfraction enriched in FA with two double bonds extracted from the gonads was obtained by using argentation thin-layer chromatography. The resulting fraction was analyzed by using capillary gas chromatography-mass spectrometry of its methyl esters, 3-pyridylcarbinol esters and pyrrolidide derivatives. Six novel all-cis diene isomers were identified as 7,18-heneicosadienoic (21:2Δ7,18), 8,14-tricosadienoic (23:2Δ8,14), 9,15-tricosadienoic (23:2Δ9,15), 7,18-tricosadienoic (23:2Δ7,18), 9,18-tricosadienoic (23:2Δ9,18), and 9,20-tricosadienoic (23:2Δ9,20) acids. In the present study, the differences in the proportion of tricosadienoic acid isomers between the ovary lipids of C. grata and C. toreuma were recognized.     

Alumina–carbon composite fibers from poly[(acyloxy)aloxane]s

The alumina–carbon composite fibers were obtained from poly[(acyloxy)aloxane] (PAA) with 3-ethoxypropanoic (EPA) and m-anisic acids (m-AA) legands. This preceramic polymer can be dissolved in p-xylene-methanol-EPA mixed solvent, and the concentrated solution exhibited an excellent spinnability. During the pyrolysis and sintering processes, aliphatic carboxylate in the side groups was easily decomposed and eliminated. The aromatic carboxylate, however, seems to be converted and migrated to a carbon domain in the alumina matrix into which aloxane repetition was converted. The fibers pyrolyzed up to 800 and 1000°C have electrical conductivities that monotonically increase with increasing temperature. The fiber pyrolyzed up to 1200°C showed the electrical conductivity in a rather complicated manner.     

Effect of acid treatment on thermal extraction yield in ashless coal production

Coals of different ranks were acid-treated in aqueous methoxyethoxy acetic acid (MEAA), acetic acid (AA), and HCl. The acid-treated coals were extracted with polar N-methyl-2-pyrrolidinone (NMP) and nonpolar 1-methylnaphthalene (1MN) solvents at temperatures from 200 to 360 °C for 10-60 min. The thermal extraction yields with NMP for some acid-treated low-rank coals increased greatly; for example, the extraction yield for Wyodak coal (%C; 75.0%) increased from 58.4% for the raw coal to 82.9% for coal treated in 1.0 M MEAA. Conversely, the extraction yields changed minimally for all the acid-treated coals extracted in 1-MN. The type and concentration of acid affected the extraction yield when NMP was used as the extraction solvent. With increasing MEAA concentration from 0.01 to 0.1 M, the extraction yield for Wyodak coal increased from 66.3 to 81.4%, and subsequently did not change clearly with concentration. Similar changes in the extraction yield with acid concentration were also observed with AA and HCl. The de-ashing ratio for coals acid-treated in MEAA, AA, and HCl also increased greatly with concentration from 0.01 to 0.1 M, which corresponded to the change in the thermal extraction yield in NMP. For the acid-treated coals, high extraction yields were obtained at lower extraction temperatures and shorter extraction times than for the raw coal. The mechanisms for the acid treatment and thermal extraction are discussed.     

Effect of a low-fat diet enriched with oleic acid on postprandial lipemia in patients with type 2 diabetes mellitus

The aim of the present study was to compare the effects of a low-fat diet enriched with oleic acid to those of a low-fat diet enriched with linoleic acid on fasting lipids, postprandial lipemia, and oxidative susceptibility of low-density lipoprotein (LDL) in patients with type 2 diabetes mellitus (DM). In a 3-wk randomized crossover study, eight patients with type 2 DM were given an experimental low-fat diet enriched with either oleic acid or linoleic acid. The oleic-acid-enriched diet contained 5, 15, and 5% energy from saturated, monounsaturated, and polyunsaturated fatty acids, and the linoleic-acid-enriched diet contained 5, 5, and 15% energy from saturated, monounsaturated, and polyunsaturated fatty acids, respectively. In addition to evaluating the fasting lipids and oxidative susceptibility of LDL, we evaluated postprandial lipemia using an oral fat load at the end of each 3-wk dietary phase. There were no significant differences in fasting lipid profile or lag time of LDL oxidation between the two experimental dietary phases. The average and maximal increments of remnant-like particle (RLP) cholesterol levels during oral fat load were significantly higher after the oleic-acid-enriched dietary phase than after the linoleic-acid-enriched dietary phase. The area under the curve of RLP cholesterol was also significantly larger after the oleicacid-enriched dietary phase than after the linoleic-acid-enriched dietary phase. These results suggest that the oleic-acidenriched diet was associated with increased formation of postprandial chylomicron remnants compared with the linoleicacid-enriched diet.     

Centrifugal Filter for Aerosol Collection

Air filters collect particles by the mechanical collection mechanisms, namely, inertia, interception, gravitational settling, and Brownian diffusion. There exists the most penetrating particle size (MPPS) in submicron size range for which none of the collection mechanisms work effectively. In this study, we propose a new type of filter named as “centrifugal filter,” which collects aerosol particles by centrifugal force together with the conventional mechanical collection mechanisms. The centrifugal filter proposed in the present work may be rotated by a motor or compressed air. Air passes through the filter in the axial direction of filter rotation. The filter rotates so does the air embedded in the filter, and therefore centrifugal force exerts on particles. In addition to the mechanical collection mechanisms, small migration of particles due to the centrifugal force enhanced the collection efficiency of submicron particles significantly without increasing the pressure drop. The performance tests of centrifugal filter were conducted by changing the fiber diameter, the air flow velocity and the rotation speed. We found that the collection efficiency of filter is enhanced significantly by rotating the filter without increasing the pressure drop and that the filter efficiency is well predicted by the conventional filtration theory accounting for the centrifugal force.

Copyright 2015 American Association for Aerosol Research     

Immunological Effects of Oenothein B,an Ellagitannin Dimer,on Dendritic Cells

Oenothein B is a unique macrocyclic ellagitannin dimer that has been found in various medicinal plants belonging to Onagraceae, Lythraceae, and Myrtaceae, with diverse biological activities. The immunological effects of tannins in terms of cytokine-release from macrophages and monocytes have been discussed, while the effects on other immunocompetent cells have been the subject of minimal investigation. We evaluated the immunomodulatory effects induced by tannin treatment in human dendritic cells (DCs), which play a critical role in the initial immune response, by measuring the changes in cytokine production, cell differentiation, and cell viability. Oenothein B showed significant down-regulation of the expression of cell surface molecules, CD1a and CD83, suggesting the inhibition of DC differentiation and/or maturation. The suppressive effect on DCs was associated with the induction of apoptosis without the activation of caspase-3/7, 8, and 9, and this was supported by the morphological features indicating significant nuclear condensation. Oenothein B also markedly suppressed the production of inflammatory cytokines, such as IL-1β and IL-6, in a dose-dependent manner. These data may, in part, be able to explain the traditional use of tannin-containing medicinal plants for the treatment of a variety of inflammatory diseases, including inflammatory bowel disease, celiac disease, and rheumatoid arthritis.     

Effect of helix-coil transition on association behavior of both-ends hydrophobically-modified water-soluble polypeptide

Dodecyl and dodecanoyl groups (C12) are attached at the chain ends of poly[N⁵-(2-hydroxyethyl) l-glutamine] (PHEG), and association behavior of this both-ends hydrophobically modified water soluble polypeptide (C12-PHEG-C12) has been investigated by means of light scattering measurements. Water/ethylene glycol (EG) mixed solvents were used as selective solvent for PHEG block, and PHEG changed its structure from random-coil state to α-helix with increasing EG content in the mixed solvent (W_EG). When W_EG is less than 0.5, flower-like micelle with C12 associated core and PHEG corona in loop conformation was suggested to be formed. Increase of W_EG from 0.5 to 0.6 induced drastic increase of association number and size of the associate, in which many C12 associated cores may be connected by PHEG in bridge conformation. This structure change of associate is considered to be driven by the increase of helix content of PHEG with W_EG, which enhances the possibility to form bridge conformation because of its rigidity. Solution preparation method, i.e. order of addition of solvent, was found to influence the structure of associate, although its effect on the helix content of PHEG was negligible.     

Differences in Sterol Composition between Male and Female Gonads of Dominant Limpet Species

The present study demonstrated here for the first time that there are statistically significant differences in sterol composition between male and female gonads of the dominant limpets Cellana grata and Cellana toreuma, which are intertidal gastropods. Among 11 different sterols identified in this study, unusually high levels (11.2–19.8% of total sterols) of the Δ⁸-sterols 5α-cholest-8-en-3β-ol (zymostenol) and 5α-cholesta-8,24-dien-3β-ol (zymosterol), which have never been reported in aquatic invertebrate gonads, were present in only the male gonads.     

Water Vapor Adsorption and Desorption on Materials Hydrothermally Solidified from Clay Minerals

Water vapor isotherms were examined and ¹H-magic-angle spinning nuclear magnetic resonance (¹H-MAS NMR) analysis was conducted to determine the amount of water vapor adsorbed and desorbed on hydrothermally solidified materials derived from clay minerals. The amount of water vapor adsorbed and desorbed decreased with increasing mesopore volume of the materials, which disagreed with the estimation obtained using the Kelvin equation. Water vapor was physisorbed on alumina-like sites in the materials. An increase in the binding energy between water vapor and the materials led to an improvement in the water vapor adsorption–desorption properties of materials.