Ferocious Fighting between Male Grasshoppers

Contests among individuals over mating opportunities are common across diverse taxa, yet physical conflict is relatively rare. Due to the potentially fatal consequences of physical fighting, most animals employ mechanisms of conflict resolution involving signalling and ritualistic assessment. Here we provide the first evidence of ubiquitous escalated fighting in grasshoppers. The chameleon grasshopper (Kosciuscola tristis) is an Australian alpine specialist, in which males engage in highly aggressive combat over ovipositing females. We describe discrete agonistic behaviours including mandible flaring, mounting, grappling, kicking and biting, and their use depending on the individual’s role as challenger or defender. We show that male role predicts damage, with challengers being more heavily damaged than males defending females (defenders). Challengers also possess wider mandibles than defenders, but are similar in other metrics of body size. Our data suggest that fights escalate between males matched in body size and that mandibles are used as weapons in this species. This system represents an exciting opportunity for future research into the evolution of costly fighting behaviour in an otherwise placid group.     

High-level expression and characterization of a highly functional Comamonas acidovorans xanthine dehydrogenase in Pseudomonas aeruginosa

An improved procedure is described for the high-level expression of Comamonas acidovorans XDH in Pseudomonas aeruginosa PAO1-LAC. The level of functional expression (56 mg protein/L culture) is found to be 7-fold higher than that observed in Escherichia coli and 30-fold higher than that induced in C. acidovorans. Co-expression of the xdhC gene is required for maximal level of functional expression. Comparison of purified preparations of XDH expressed in the absence of xdhC (XDH(AB)) with that expressed in its presence (XDH(ABC)) shows the increased level of activity due to the level of Mo incorporation. The Fe and FAD contents of expressed enzymes are independent of xdhC co-expression. Electron paramagnetic resonance spectroscopy, circular dichroism spectroscopy, metal analysis, and kinetic properties of recombinant purified XDH(ABC) are identical with those exhibited by the native enzyme. This expression system should serve as a valuable tool for further biophysical and mechanistic investigations of xanthine dehydrogenase by site-directed mutagenesis. A method is also described to evaluate the suitability of P. aeruginosa and other organisms as potential expression hosts for five different sources of xdh genes.     

RNA-hydrolyzing activity of human serum albumin and its recombinant analogue

The comparative analysis of RNA-hydrolyzing activity of albumin from human serum and albumin expressed in methylotrophic yeast Pichia pastoris has been carried out. The rate of polyribonucleotide phosphodiester bond cleavage in the presence of recombinant albumin has been found to be similar to that of the reaction mediated by the native protein. According to ³¹P NMR data, RNA hydrolysis follows the mechanism of intermolecular trans-esterification to yield 2′,3′-cyclophosphodiester reaction products that are further slowly hydrolyzed to form nucleoside-3′- and nucleoside-2′-phosphates. Analysis of pH dependence suggests an acid–base mechanism of catalysis. The catalytic activity and substrate specificity of albumin in RNA hydrolysis distinguish it from human ribonucleases.     

Graded Effects of Social Conformity on Recognition Memory

Previous studies have shown that the opinion of confederates in a group influences recognition memory, but inconsistent results have been obtained concerning the question of whether recognition of items as old and new are affected similarly, possibly because only one or two confederates are present during the recognition phase. Here, we present data from a study where recognition of novel faces was tested in the presence of four confederates. In a long version of this experiment, recognition of items as old and new was similarly affected by group responses. However, in the short version, recognition of old items depended proportionally on the number of correct group responses, while rejection of new items only decreased significantly when all confederates gave an incorrect response. These findings indicate that differential effects of social conformity on recognition of items as old and new occur in situations with an intermediate level of group pressure.     

Effects of temperature regime on primary producers in Icelandic geothermal streams

Eight adjacent sub-arctic streams with consistently different temperatures but broadly similar chemistry, other than some differences in conductivity and linked pH, were used to investigate effects of temperature regime on the structure of primary producer communities. Grazing by invertebrates was also taken into account to detect possible effects on the primary producers.The moss species Fontinalis antipyretica Hedw. (Bryophyta) was only observed in the warmer streams and its cover was positively linked with temperature. The liverwort Jungermannia exsertifolia (Steph.) was absent from the warmer streams and scarce in the colder streams. The biomass of F. antipyretica was inversely correlated with concentration of epilithic chlorophyll a, possibly due to shading from the bryophyte and/or competition between algae and bryophytes for nutrients. Epilithic algae and bryophyte biomass were not related to density of the main grazer, Radix peregra Müller (Gastropoda).A high biovolume of green algae (Chlorophyta) occurred in all streams. The biovolume Cyanobacteria was greatest in the coldest stream and in the two intermediately warm streams. Diatom biovolume was low compared with the biovolumes of Cyanobacteria and green algae in all streams, except in the two warmest streams, where the highest biovolume of diatom occurred.Regularized Canonical Correlation Analysis (RCCA) classification based on all primary producer assemblages and invertebrates indicated that warm streams (16.7–22.2 °C) were similar to each other. It also showed that the colder streams (6.6–13.2 °C) were different from the warmer streams and from each other because of differences in conductivity. Temperature and conductivity were the most influential variables in determining overall stream diversity.     

Bioluminescent imaging of Cdk2 inhibition in vivo

Many proteins and pathways of pharmaceutical interest impinge on ubiquitin ligases or their substrates. The cyclin-dependent kinase (Cdk) inhibitor p27, for example, is polyubiquitylated in a cell cycle-dependent manner by a ubiquitin ligase complex containing the F-box protein Skp2. Regulated turnover of p27 is due, at least partly, to its phosphorylation by Cdk2 on threonine 187, which generates a Skp2-binding site. We made a p27-luciferase (p27Luc) fusion protein and show here that its abundance, like that of p27, is regulated by Skp2 in a cell cycle-dependent manner. As predicted, p27Luc levels increased after blocking Cdk2 activity with inhibitory proteins, peptides or small interfering RNA (siRNA). Accumulation of p27Luc in response to Cdk2 inhibitory drugs (flavopiridol and R-roscovitine) was demonstrable in human tumor cells in vivo using noninvasive bioluminescent imaging. In theory, the approach described here could be used to develop bioluminescent reporters for any drug target that directly or indirectly affects the turnover of a ubiquitin ligase substrate.     

Inclusion complexes of carotenoids with cyclodextrins: 1H NMR, EPR, and optical studies

Direct evidence of carotenoid/cyclodextrin inclusion complex formation was obtained for the water-soluble sodium salt of beta-caroten-8'-oic acid (IV) by using 1H NMR and UV-Vis absorption spectroscopy. It was shown that this carotenoid forms a stable 1:1 inclusion complex with beta-cyclodextrin (stability constant K11 approximately 1500 M(-1)). All other carotenoids under study in the presence of cyclodextrins (CDs) form large aggregates in aqueous solution as demonstrated by very broad absorption spectra and considerable change in color. By using the EPR spin trapping technique, the scavenging ability of IV toward OOH radicals was compared in DMSO and in the aqueous CD solution. A considerable decrease in PBN/OOH spin adduct yield was detected in the presence of uncomplexed IV because of a competing reaction of the carotenoid with OOH radical. No such decrease occurred in the presence of the IV/CD complex. Moreover, a small increase in spin adduct yield (pro-oxidant effect) is most likely due to the reaction of the carotenoid with Fe3+ to regenerate Fe2+, which in turn regenerates the OOH radical. Our data show that CD protects the carotenoid from reactive oxygen species. On the other hand, complexation with CD results in considerable decrease in antioxidant ability of the carotenoid.     

The purple Trp288Ala mutant of Synechocystis OCP persistently quenches phycobilisome fluorescence and tightly interacts with FRP

In Cyanobacteria, the Orange Carotenoid Protein (OCP) and Fluorescence Recovery Protein (FRP) are central to the photoprotective mechanism consisting in regulated quenching of phycobilisome (PBs) fluorescence. Due to a transient and flexible nature of the light-activated red quenching form, OCP^R, which is obtained from the stable dark-adapted orange form, OCP^O, by photoconversion, the detailed mechanism of photoprotection remains unclear. Here we demonstrate that our recently described W288A mutant of the Synechocystis OCP (hereinafter called OCP^W288A) is a fully functional analogue of the OCP^R form which is capable of constitutive PBs fluorescence quenching in vitro with no need of photoactivation. This PBs quenching effect is abolished in the presence of FRP, which interacts with OCP^W288A with micromolar affinity and an apparent stoichiometry of 1:1, unexpectedly, implying dissociation of the FRP dimers. This establishes OCP^W288A as a robust model system providing novel insights into the interplay between OCP and FRP to regulate photoprotection in cyanobacteria.