Modulating cholesteryl ester transfer protein activity maintains efficient pre-��-HDL formation and increases reverse cholesterol transport

The mechanism by which cholesteryl ester transfer protein (CETP) activity affects HDL metabolism was investigated using agents that selectively target CETP (dalcetrapib, torcetrapib, anacetrapib). In contrast with torcetrapib and anacetrapib, dalcetrapib requires cysteine 13 to decrease CETP activity, measured as transfer of cholesteryl ester (CE) from HDL to LDL, and does not affect transfer of CE from HDL3 to HDL2. Only dalcetrapib induced a conformational change in CETP, when added to human plasma in vitro, also observed in vivo and correlated with CETP activity. CETP-induced pre-β-HDL formation in vitro in human plasma was unchanged by dalcetrapib ≤3 µM and increased at 10 µM. A dose-dependent inhibition of pre-β-HDL formation by torcetrapib and anacetrapib (0.1 to 10 µM) suggested that dalcetrapib modulates CETP activity. In hamsters injected with [³H]cholesterol-labeled autologous macrophages, and given dalcetrapib (100 mg twice daily), torcetrapib [30 mg once daily (QD)], or anacetrapib (30 mg QD), only dalcetrapib significantly increased fecal elimination of both [³H]neutral sterols and [³H]bile acids, whereas all compounds increased plasma HDL-[³H]cholesterol. These data suggest that modulation of CETP activity by dalcetrapib does not inhibit CETP-induced pre-β-HDL formation, which may be required to increase reverse cholesterol transport.     

<Emphasis Type="Italic">AtMRP6</Emphasis>/<Emphasis Type="Italic">AtABCC6</Emphasis>, an ATP-Binding Cassette transporter gene expressed during early steps of seedling development and up-regulated by cadmium in <Emphasis Type="Italic">Arabidopsis thaliana</Emphasis>

Background  

ABC proteins constitute one of the largest families of transporters found in all living organisms. In Arabidopsis thaliana, 120 genes encoding ABC transporters have been identified. Here, the characterization of one member of the MRP subclass, AtMRP6, is described.     

Long-term evolutionary stability of bacterial endosymbiosis in curculionoidea: additional evidence of symbiont replacement in the dryophthoridae family

Bacterial intracellular symbiosis (endosymbiosis) is well documentedin the insect world where it is believed to play a crucial rolein adaptation and evolution. However, although Coleopteran insectsare of huge ecological and economical interest, endosymbiontmolecular analysis is limited to the Dryophthoridae family.Here, we have analyzed the intracellular symbiotic bacteriain 2 Hylobius species belonging to the Molytinae subfamily (Curculionoideasuperfamily) that exhibit different features from the Dryophthoridaeinsects in terms of their ecology and geographical spanning.Fluorescence in situ hybridization has shown that both Hylobiusspecies harbor rod-shaped pleiomorphic symbiotic bacteria inthe oocyte and in the bacteria-bearing organ (the bacteriome),with a shape and location similar to those of the Dryophthoridaebacteriome. Phylogenetic analysis of the 16S ribosomal DNA genesequences, using the heterogeneous model of DNA evolution, hasplaced the Hylobius spp. endosymbionts (H-group) at the basalposition of the ancestral R-clade of Dryophthoridae endosymbiontsnamed Candidatus Nardonella but relatively distant from theS-clade of Sitophilus spp. endosymbionts. Endosymbionts fromthe H-group and the R-clade evolved more quickly compared withfree-living enteric bacteria and endosymbionts from the S- andD-clades of Dryophthoridae. They are AT biased (58.3% A + T),and they exhibit AT-rich insertions at the same position aspreviously described in the Candidatus Nardonella 16S rDNA sequence.Moreover, the host phylogenetic tree based on the mitochondrialCOI gene was shown to be highly congruent with the H-group andthe R-clade, the divergence of which was estimated to be around125 MYA. These new molecular data show that endosymbiosis isold in Curculionids, going back at least to the common ancestorof Molytinae and Dryophthoridae, and is evolutionary stable,except in 2 Dryophthoridae clades, providing additional andindependent supplementary evidence for endosymbiont replacementin these taxa.     

Selective cell adhesion inhibitors: Barbituric acid based alpha4beta7--MAdCAM inhibitors

A novel series of barbituric acid derivatives were identified as selective inhibitors of alpha4beta7 MAdCAM (mucosal addressin cell adhesion molecule-1) interactions via a high throughput screening exercise. These inhibitors were optimized to submicromolar potencies in whole cell adhesion assays, retaining their selectivity over alpha4beta1 VCAM.     

Synthesis and biological evaluation of bengacarboline derivatives

Novel derivatives of the marine alkaloid bengacarboline have been synthesized. The seco derivatives 11 and 12 were evaluated for topoisomerase inhibition, DNA damages, cytotoxicity and cell cycle perturbation. The two synthetic analogs are more potent cytotoxic agents than bengacarboline and they both induce an accumulation of cells in the S phase of DNA synthesis. They do not function as topoisomerase inhibitors but trigger DNA damages in cells.     

THE EFFECT OF VESSEL NOISE ON THE VOCAL BEHAVIOR OF BELUGAS IN THE ST. LAWRENCE RIVER ESTUARY,CANADA

During June-July 1991, we monitored the vocal behavior of belugas before, during, and after exposure to noise from a small motorboat and a ferry to determine if there were any consistent patterns in their vocal behavior when exposed to these two familiar, but different sources of potential disturbance. Vocal responses were observed in all trials and were more persistent when whales were exposed to the ferry than to the small boat. These included (1) a progressive reduction in calling rate from 3.4–10.5 calls/whale/min to 0.0 or <1.0 calls/whale/min while vessels were approaching; (2) brief increases in the emission of falling tonal calls and the theree pulsed-tone call types; (3) at distances <1 km, an increase in the repetition of specific calls, and (4) a shift in frequency bands used by vocalizing animals from a mean frequency of 3.6 kHz prior to exposure to noise to frequencies of 5.2-8.8 kHz when vessels were close to the whales.     

Molecular modeling and site-directed mutagenesis of plant chloroplast monogalactosyldiacylglycerol synthase reveal critical residues for activity

Monogalactosyldiacylglycerol (MGDG), the major lipid of plant and algal plastids, is synthesized by MGD (or MGDG synthase), a dimeric and membrane-bound glycosyltransferase of the plastid envelope that catalyzes the transfer of a galactosyl group from a UDP-galactose donor onto a diacylglycerol acceptor. Although this enzyme is essential for biogenesis, and therefore an interesting target for herbicide design, no structural information is available. MGD monomers share sequence similarity with MURG, a bacterial glycosyltransferase catalyzing the transfer of N-acetyl-glucosamine on Lipid 1. Using the x-ray structure of Escherichia coli MURG as a template, we computed a model for the fold of Spinacia oleracea MGD. This structural prediction was supported by site-directed mutagenesis analyses. The predicted monomer architecture is a double Rossmann fold. The binding site for UDP-galactose was predicted in the cleft separating the two Rossmann folds. Two short segments of MGD (beta2-alpha2 and beta6-beta7 loops) have no counterparts in MURG, and their structure could not be determined. Combining the obtained model with phylogenetic and biochemical information, we collected evidence supporting the beta2-alpha2 loop in the N-domain as likely to be involved in diacylglycerol binding. Additionally, the monotopic insertion of MGD in one membrane leaflet of the plastid envelope occurs very likely at the level of hydrophobic amino acids of the N-terminal domain.     

Effects of nonlipolytic ligand function of endothelial lipase on high density lipoprotein metabolism in vivo

Endothelial lipase (EL) influences high density lipoprotein (HDL) metabolism in vivo and mediates bridging and uptake of HDL particles independent of its lipolytic activity in vitro. To determine whether EL has a nonlipolytic ligand function in HDL metabolism in vivo, 1 x 1011 particles of a recombinant adenovirus encoding human EL (AdEL), catalytically inactive human EL (AdELS149A), or control (Adnull) were injected into wild-type, apoA-I transgenic, and hepatic lipase knockout mice. ELS149A protein was expressed at higher levels than wild-type EL. EL and ELS149A protein were both substantially increased in the postheparin plasma compared with preheparin, indicating that both the wild-type and mutant EL were bound to cell-surface heparan sulfate proteoglycans. Overexpression of wild-type EL was associated with a significantly increased postheparin-plasma phospholipase activity and dramatically decreased levels of total cholesterol, HDL cholesterol, phospholipids, and apoA-I. Injection of AdELS149A did not result in increased phospholipase activity confirming that ELS149A was catalytically inactive. Expression of ELS149A did not decrease lipid or apoA-I levels in wild-type and apoA-I transgenic mice yet led to an intermediate reduction of total cholesterol, HDL cholesterol, and phospholipids in hepatic lipase-deficient mice compared with control and EL-expressing mice. Our study demonstrates for the first time that EL has both a lipolytic and nonlipolytic function in HDL metabolism in vivo. Lipolytic activity of EL, however, seems to be most important for its effects on systemic HDL metabolism.