Discovery and Targeted LC-MS/MS of Purified Polerovirus Reveals Differences in the Virus-Host Interactome Associated with Altered Aphid Transmission

Circulative transmission of viruses in the Luteoviridae, such as cereal yellow dwarf virus (CYDV), requires a series of precisely orchestrated interactions between virus, plant, and aphid proteins. Natural selection has favored these viruses to be retained in the phloem to facilitate acquisition and transmission by aphids. We show that treatment of infected oat tissue homogenate with sodium sulfite reduces transmission of the purified virus by aphids. Transmission electron microscopy data indicated no gross change in virion morphology due to treatments. However, treated virions were not acquired by aphids through the hindgut epithelial cells and were not transmitted when injected directly into the hemocoel. Analysis of virus preparations using nanoflow liquid chromatography coupled to tandem mass spectrometry revealed a number of host plant proteins co-purifying with viruses, some of which were lost following sodium sulfite treatment. Using targeted mass spectrometry, we show data suggesting that several of the virus-associated host plant proteins accumulated to higher levels in aphids that were fed on CYDV-infected plants compared to healthy plants. We propose two hypotheses to explain these observations, and these are not mutually exclusive: (a) that sodium sulfite treatment disrupts critical virion-host protein interactions required for aphid transmission, or (b) that host infection with CYDV modulates phloem protein expression in a way that is favorable for virus uptake by aphids. Importantly, the genes coding for the plant proteins associated with virus may be examined as targets in breeding cereal crops for new modes of virus resistance that disrupt phloem-virus or aphid-virus interactions.     

N-Benzyl-N-(pyrrolidin-3-yl)carboxamides as a new class of selective dual serotonin/noradrenaline reuptake inhibitors

The structure–activity relationship and the synthesis of novel N-benzyl-N-(pyrrolidin-3-yl)carboxamides as dual serotonin (5-HT) and noradrenaline (NA) monoamine reuptake inhibitors are described. Compounds such as 18 exhibited dual 5-HT and NA reuptake inhibition, good selectivity over dopamine (DA) reuptake inhibition and drug-like physicochemical properties consistent with CNS target space. Compound 18 was selected for further preclinical evaluation.     

Passive cambering and flexible propulsors: cetacean flukes

The flukes are the primary locomotor structure in cetaceans, which produce hydrodynamic thrust as the caudal vertebrae are oscillated dorso-ventrally. Effective thrust generation is a function of the kinematics of the flukes, the angle of attack between the flukes and the incident water flow, and the shape of the flukes. We investigated the effect of bending within the caudal region of odontocete cetaceans to determine how changes in angular displacement between caudal vertebrae could effect passive shape change of the flukes. The internal and external changes of bent flukes were examined with computer tomography. Flukes and tailstock were removed from deceased Delphinus delphis, Lagenorhynchus acutus, Peponocephala electra, Phocoena phocoena and Tursiops truncatus, and bent on an adjustable support at 0, 45 and 90 degrees . At 0 degrees , cross-sections of the flukes displayed a symmetrical profile. Cross-sections of bent flukes (45 degrees , 90 degrees ) were asymmetrical and showed a cambered profile. Maximum cambering occurred close to the tailstock and decreased toward the fluke tip. Maximum angular displacement occurred at the 'ball vertebra', which was located posterior of the anterior insertion of the flukes on the tailstock. Bending at the 'ball vertebra' passively cambers the flexible flukes. Cambering could increase hydrodynamic force production during swimming, particularly during direction reversal in the oscillatory cycle.     

6,7-Dihydro-5H-pyrrolo[1,2-a] imidazoles as potent and selective α1A adrenoceptor partial agonists

Novel pyrroloimidazoles have been identified as potent partial agonists of the α_1A adrenergic receptor, with good selectivity over the α_1B, α_1D and α_2A receptor subtypes. Pyrimidine 19 possessed attractive CNS drug-like properties with good membrane permeability and no evidence for P-gp mediated efflux.     

Meta-QTL analysis of the genetic control of ear emergence in elite European winter wheat germplasm

Variation in ear emergence time is critical for the adaptation of wheat (Triticum aestivum L.) to specific environments. The aim of this study was to identify genes controlling ear emergence time in elite European winter wheat germplasm. Four doubled haploid populations derived from the crosses: Avalon × Cadenza, Savannah × Rialto, Spark × Rialto, and Charger × Badger were selected which represent diversity in European winter wheat breeding programmes. Ear emergence time was recorded as the time from 1st May to heading in replicated field trials in the UK, France and Germany. Genetic maps based on simple sequence repeat (SSR) and Diversity Arrays Technology (DArT) markers were constructed for each population. One hundred and twenty-seven significant QTL were identified in the four populations. These effects were condensed into 19 meta-QTL projected onto a consensus SSR map of wheat. These effects are located on chromosomes 1B (2 meta-QTL), 1D, 2A (2 meta-QTL), 3A, 3B (2 meta-QTL), 4B, 4D, 5A (2 meta-QTL), 5B, 6A, 6B 7A (2 meta-QTL), 7B and 7D. The identification of environmentally robust earliness per se effects will facilitate the fine tuning of ear emergence in predictive wheat breeding programmes. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

Nature and Recurrence of AVPR2 Mutations in X-linked Nephrogenic Diabetes Insipidus

X-linked nephrogenic diabetes insipidus (NDI) is a rare disease with defective renal and extrarenal arginine-vasopressin V₂ receptor responses due to mutations in the AVPR2 gene in Xq28. We analyzed 31 independent NDI families to determine the nature and recurrence of AVPR2 mutations. Twenty-one new putative disease-causing mutations were identified: 113delCT, 253del35, 255del9, 274insG, V88M, R106C, 402delCT, C112R, Y124X, S126F, W164S, S167L, 684delTA, 804insG, W284X, A285P, W293X, R337X, and three large deletions or gene rearrangements. Five other mutations—R113W, Y128S, R137H, R181C, and R202C—that previously had been reported in other families were detected. There was evidence for recurrent mutation for four mutations (R113W, R137H, S167L, and R337X). Eight de novo mutation events were detected (274insG, R106C, Y128S, 167L [twice], R202C, 684delTA, and R337X). The origins were maternal (one), grandmaternal (one), and grandpaternal (six). In the 31 NDI families and 6 families previously reported by us, there is evidence both for mutation hot spots for nucleotide substitutions and for small deletions and insertions. More than half (58%) of the nucleotide substitutions in 26 families could be a consequence of 5-methylcytosine deamination at a CpG dinucleotide. Most of the small deletions and insertions could be attributed to slipped mispairing during DNA replication.     

Patterns of fungal diversity and composition along a salinity gradient

Estuarine salinity gradients are known to influence plant, bacterial and archaeal community structure. We sequenced 18S rRNA genes to investigate patterns in sediment fungal diversity (richness and evenness of taxa) and composition (taxonomic and phylogenetic) along an estuarine salinity gradient. We sampled three marshes—a salt, brackish and freshwater marsh—in Rhode Island. To compare the relative effect of the salinity gradient with that of plants, we sampled fungi in plots with Spartina patens and in plots from which plants were removed 2 years prior to sampling. The fungal sediment community was unique compared with previously sampled fungal communities; we detected more Ascomycota (78%), fewer Basidiomycota (6%) and more fungi from basal lineages (16%) (Chytridiomycota, Glomeromycota and four additional groups) than typically found in soil. Across marshes, fungal composition changed substantially, whereas fungal diversity differed only at the finest level of genetic resolution, and was highest in the intermediate, brackish marsh. In contrast, the presence of plants had a highly significant effect on fungal diversity at all levels of genetic resolution, but less of an effect on fungal composition. These results suggest that salinity (or other covarying parameters) selects for a distinctive fungal composition, and plants provide additional niches upon which taxa within these communities can specialize and coexist. Given the number of sequences from basal fungal lineages, the study also suggests that further sampling of estuarine sediments may help in understanding early fungal evolution.     

Characterisation of vortex shedding in vascular anastomosis models using pulsed Doppler ultrasound.

Vortex shedding at vascular anastomoses were investigated in vitro using a 20 MHz pulsed-wave Doppler velocimeter. Centreline velocity measurements were made at various axial distances in simplified polyurethane models of proximal and distal end-to-side anastomoses of angles 15, 30, 45, 60 and 80 degrees using pulsatile flow waveforms similar to those in femoropopliteal bypass grafts. The in-phase and quadrature Doppler signals were recorded and the maximum frequency waveform, averaged over 64 cycles, was obtained using short-time Fourier transform. A fourth-order Butterworth low-pass filter was employed to separate the vortex velocity signal from the convective velocity. The vortex signal envelope was calculated using a Hilbert transform method and the vortex amplitude was taken as the maximum of this envelope. The results show that higher vortex amplitude were found in the proximal anastomoses and under resting flow conditions. Although the vortex amplitudes generally increased with angles of anastomosis, they were found to be higher in the 60 degrees than in the 80 degrees proximal anastomosis. The vortex structures were investigated using spectrograms and these show prominent features at 40-50 Hz indicative of the short-duration oscillatory signals during the decelerative phase of systole expected from the passage of vortices. The study indicates that flow disturbances due to vortex shedding may be a common feature in femoropopliteal bypass grafts.