Parasitism and mutualism in Wolbachia: what the phylogenomic trees can and cannot say

Ecological and evolutionary theories predict that parasitismand mutualism are not fixed endpoints of the symbiotic spectrum.Rather, parasitism and mutualism may be host or environmentdependent, induced by the same genetic machinery, and shifteddue to selection. These models presume the existence of geneticor environmental variation that can spur incipient changes insymbiotic lifestyle. However, for obligate intracellular bacteriawhose genomes are highly reduced, studies specify that discretesymbiotic associations can be evolutionarily stable for hundredsof millions of years. Wolbachia is an inherited obligate, intracellularinfection of invertebrates containing taxa that act broadlyas both parasites in arthropods and mutualists in certain roundworms.Here, we analyze the ancestry of mutualism and parasitism inWolbachia and the evolutionary trajectory of this variationin symbiotic lifestyle with a comprehensive, phylogenomic analysis.Contrary to previous claims, we show unequivocally that thetransition in lifestyle cannot be reconstructed with currentmethods due to long-branch attraction (LBA) artifacts of thedistant Anaplasma and Ehrlichia outgroups. Despite the use of1) site-heterogenous phylogenomic methods that can overcomesystematic error, 2) a taxonomically rich set of taxa, and 3)statistical assessments of the genes, tree topologies, and modelsof evolution, we conclude that the LBA artifact is serious enoughto afflict past and recent claims including the root lies inthe middle of the Wolbachia mutualists and parasites. We showthat different inference methods yield different results andhigh bootstrap support did not equal phylogenetic accuracy.Recombination was rare among this taxonomically diverse dataset, indicating that elevated levels of recombination in Wolbachiaare restricted to specific coinfecting groups. In conclusion,we attribute the inability to root the tree to rate heterogeneitybetween the ingroup and outgroup. Site-heterogenous models ofevolution did improve the placement of aberrant taxa in theingroup phylogeny. Finally, in the unrooted topology, the distributionof parasitism and mutualism across the tree suggests that atleast two interphylum transfers shaped the origins of nematodemutualism and arthropod parasitism. We suggest that the ancestryof mutualism and parasitism is not resolvable without more suitableoutgroups or complete genome sequences from all Wolbachia supergroups.     

Design,synthesis and biological evaluation of cyclic angiotensin II analogues with 3,5 side-chain bridges. Role of C-terminal aromatic residue and ring cluster for activity and implications in the drug design of AT1 non-peptide antagonists

The novel amide linked angiotensin II (ANG II) cyclic analogues: gamma, epsilon -cyclo(3, 5)-[Sar(1)-Glu(3)-Lys(5)-Ile(8)] ANG II (I) and gamma, epsilon -cyclo(3, 5)-[Sar(1)-Glu(3)-Lys(5)-Phe(8)] ANG II (II) have been designed, synthesized and bioassayed in anesthetized rabbits in order to unravel structural ring cluster characteristics important for receptor activation. Analogue I with Ile at position 8 was an inhibitor of Angiotensin II while analogue II with Phe at position 8 was found to be an agonist. Similar results were reported for cyclic compounds that have reversed the linking between positions 3 and 5. The overall results show that positions 3 and 5 do not govern the biological activity of the synthetic analogues. It also appears that the aromatic ring cluster (Tyr-His-Phe) in agonist peptides is an essential stereo-electronic feature for Angiotensin II to exert its biological activity. A non-peptide mimetic of ANG II, 1-[2'-[(N-benzyl)tetrazol-5-yl]biphenyl-4-yl]methyl]-2-hydroxymethylbenzimidazole (BZI8) has been designed and synthesized. This molecule is more rigid and much less active than AT(1) non-peptide mimetic losartan probably because it lacks to mimic the orientation of tetrazole and the pharmacophore segments of butyl chain and imidazole ring.     

Mechanisms of proteasome inhibitor-induced cytotoxicity in malignant glioma

The 26S proteasome constitutes an essential degradation apparatus involved in the consistent recycling of misfolded and damaged proteins inside cells. The aberrant activation of the proteasome has been widely observed in various types of cancers and implicated in the development and progression of carcinogenesis. In the era of targeted therapies, the clinical use of proteasome inhibitors necessitates a better understanding of the molecular mechanisms of cell death responsible for their cytotoxic action, which are reviewed here in the context of sensitization of malignant gliomas, a tumor type particularly refractory to conventional treatments.     

Synthesis and cytotoxic activity of some new azapyranoxanthenone aminoderivatives

A series of novel azapyranoxanthenones, bearing structural similarity to the acridone alkaloid acronycine have been designed and synthesized. Their in vitro cytotoxicities against the murine L1210 leukemia and the human solid tumor HT-29 cell lines have been investigated. The new derivatives exhibited interesting cytotoxic activity and were more potent than the parent compound.     

Inositol hexakisphosphate-dependent processing of Clostridium sordellii lethal toxin and Clostridium novyi alpha-toxin

Clostridium sordellii lethal toxin and Clostridium novyi α-toxin, which are virulence factors involved in the toxic shock and gas gangrene syndromes, are members of the family of clostridial glucosylating toxins. The toxins inactivate Rho/Ras proteins by glucosylation or attachment of GlcNAc (α-toxin). Here, we studied the activation of the autoproteolytic processing of the toxins by inositol hexakisphosphate (InsP(6)) and compared it with the processing of Clostridium difficile toxin B. In the presence of low concentrations of InsP(6) (<1 μM), toxin fragments consisting of the N-terminal glucosyltransferase (or GlcNAc-transferase) domains and the cysteine protease domains (CPDs) of C. sordellii lethal toxin, C. novyi α-toxin, and C. difficile toxin B were autocatalytically processed. The cleavage sites of lethal toxin (Leu-543) and α-toxin (Leu-548) and the catalytic cysteine residues (Cys-698 of lethal toxin and Cys-707 of α-toxin) were identified. Affinity of the CPDs for binding InsP(6) was determined by isothermal titration calorimetry. In contrast to full-length toxin B and α-toxin, autocatalytic cleavage and InsP(6) binding of full-length lethal toxin depended on low pH (pH 5) conditions. The data indicate that C. sordellii lethal toxin and C. novyi α-toxin are InsP(6)-dependently processed. However, full-length lethal toxin, but not its short toxin fragments consisting of the glucosyltransferase domain and the CPD, requires a pH-sensitive conformational change to allow binding of InsP(6) and subsequent processing of the toxin.     

A chassis exchange terminal to reduce truck congestion at container terminals

Truck congestion for container terminals seems to be a worldwide unwanted phenomenon. In this paper we present and analyze a chassis exchange terminal concept to reduce the congestion. The terminal works as a kind of extended gate of a group of traditional container terminals. During the night containers are collected from these container terminals using chassis (or trailers). In daytime these containers on chassis are then collected and exchanged with export containers also on chassis. By exchanging the chassis we avoid extra handling of containers. As connecting and disconnecting to a chassis can be done in a short time, the chassis exchange terminal increases handling capacity substantially during peak hours. In this paper we analyze the concept for the Maasvlakte container terminals in Rotterdam. We investigate both the effect on waiting time, as well as the environmental effects.