Synthesis and in vitro evaluation of 18F-labelled S-fluoroalkyl diarylguanidines: Novel high-affinity NMDA receptor antagonists for imaging with PET

Two S-[¹⁸F]fluoroalkylated diarylguanidines were synthesized and evaluated in vitro as potential tracers for imaging of N-methyl-d-aspartate receptors (NMDARs) with positron emission tomography (PET). [¹⁸F]1 and [¹⁸F]10 were synthesized by [¹⁸F]fluoroethylation and [¹⁸F]fluoromethylation of the thiol precursor 6, respectively. [¹⁸F]1 is a promising candidate NMDAR PET tracer, with low nanomolar affinity for the NMDA PCP-site, high selectivity and moderate lipophilicity.     

Plant volatile-induced aphid resistance in barley cultivars is related to cultivar age

Recent studies have shown that volatile chemical interaction between certain barley (Hordeum vulgare) cultivars can cause reduced host plant acceptance by the aphid Rhopalosiphum padi, and that certain cultivars can induce this effect while others can respond. In this study, we tested whether inducing and responding capabilities are linked to year of release in Swedish two-rowed spring barley. Eighteen cultivars released between 1897 and 1992 were tested in randomly selected subsets with pairwise combinations of volatile emitters and receivers. Significantly reduced aphid acceptance as a result of exposure to volatiles from plants of a different cultivar were found in 24% of the cultivar combinations. In general, older cultivars had a higher degree of aphid resistance after barley volatile treatment than did younger cultivars. The inducing effect of the emitter was also related to date of emitter cultivar release but the time relationship was reversed. Combinations with a younger volatile emitter and an older volatile receiver gave the strongest reduction in aphid acceptance of treated plants. Linear relationships between microsatellite diversity of emitting cultivars and their efficiency as inducers indicated that younger cultivars might have a more unique odour, whereas older cultivars may be more sensitive to induction.     

Wolf Movement Patterns: a Key to Estimation of Kill Rate?

Abstract: To estimate wolf (Canis lupus) kill rates from fine-scale movement patterns, we followed adult wolves in 3 territories of the Scandinavian wolf population using Global Positioning Systems (GPS) during the winters of 2001–2003. The resulting 6 datasets of 62–84 study days gave a total of 8,747 hourly GPS positions. We visited clusters of positions in the field on average 8.8 days after positioning and found moose (Alces alces) killed by wolves during the study period on 74 (8%) of the 953 clusters. The number of positions and visits to a cluster, their interaction, and the proportion of afternoon positions were significant fixed effects in mixed logistic-regression models predicting the probability of a cluster containing a wolf-killed moose. The models, however, displayed a poor goodness-of-fit and were not a suitable tool for estimating kill rates from positioning data alone. They might be used to reduce fieldwork by excluding unlikely clusters, although the reduction was not substantial. We discuss proximate factors (i.e., human disturbance and access to prey) as well as ultimate factors (i.e., social organization, intra-guild dominance, and litter size) as potential causes of the observed high temporal and spatial variation in prey-handling. For similar future kill-rate studies, we recommend increasing field efforts and shortening positioning intervals.     

Multistage,Long-Range Natal Dispersal by a Global Positioning System-Collared Scandinavian Wolf

Abstract: We document a new record dispersal for wolves worldwide. The natal straight-line dispersal distance of a Global Positioning System-collared female wolf from the Scandinavian population was 1,092 km from southeast Norway to northeast Finland, with a multistage actual travel distance of >10,000 km. Natural gene flow to the isolated, inbred Scandinavian wolf population may occur if survival of dispersers is improved.     

Phosphoproteomic screening identifies Rab GTPases as novel downstream targets of PINK1

Mutations in the PTEN‐induced kinase 1 (PINK1) are causative of autosomal recessive Parkinson''s disease (PD). We have previously reported that PINK1 is activated by mitochondrial depolarisation and phosphorylates serine 65 (Ser⁶⁵) of the ubiquitin ligase Parkin and ubiquitin to stimulate Parkin E3 ligase activity. Here, we have employed quantitative phosphoproteomics to search for novel PINK1‐dependent phosphorylation targets in HEK (human embryonic kidney) 293 cells stimulated by mitochondrial depolarisation. This led to the identification of 14,213 phosphosites from 4,499 gene products. Whilst most phosphosites were unaffected, we strikingly observed three members of a sub‐family of Rab GTPases namely Rab8A, 8B and 13 that are all phosphorylated at the highly conserved residue of serine 111 (Ser¹¹¹) in response to PINK1 activation. Using phospho‐specific antibodies raised against Ser¹¹¹ of each of the Rabs, we demonstrate that Rab Ser¹¹¹ phosphorylation occurs specifically in response to PINK1 activation and is abolished in HeLa PINK1 knockout cells and mutant PINK1 PD patient‐derived fibroblasts stimulated by mitochondrial depolarisation. We provide evidence that Rab8A GTPase Ser¹¹¹ phosphorylation is not directly regulated by PINK1 in vitro and demonstrate in cells the time course of Ser¹¹¹ phosphorylation of Rab8A, 8B and 13 is markedly delayed compared to phosphorylation of Parkin at Ser⁶⁵. We further show mechanistically that phosphorylation at Ser¹¹¹ significantly impairs Rab8A activation by its cognate guanine nucleotide exchange factor (GEF), Rabin8 (by using the Ser111Glu phosphorylation mimic). These findings provide the first evidence that PINK1 is able to regulate the phosphorylation of Rab GTPases and indicate that monitoring phosphorylation of Rab8A/8B/13 at Ser¹¹¹ may represent novel biomarkers of PINK1 activity in vivo. Our findings also suggest that disruption of Rab GTPase‐mediated signalling may represent a major mechanism in the neurodegenerative cascade of Parkinson''s disease.     

Conserved movement of TMS11 between occluded conformations of LacY and XylE of the major facilitator superfamily suggests a similar hinge‐like mechanism

The Δ‐distance maps can detect local remodeling that is difficult to accurately determine using superimpositions. Transmembrane segments (TMSs) 11 in both LacY and XylE of the major facilitator superfamily uniquely contribute the greatest amount of mobile surface area in the outward‐occluded state and undergo analogous movements. The intracellular part of TMS11 moves away from the C‐terminal domain and into the substrate cavity during the conformational change from the outward‐occluded to the inward‐occluded state. A difference was noted between LacY and XylE when they assumed the inward open state after releasing a substrate to the inside in which TMS11 of LacY moved further into the substrate release space, whereas in XylE, TMS11 slightly retracted into the C‐terminal domain. Independent movement of the N‐terminal half of TMS11 suggests that it is flexible in the middle. Repeat‐swapped homology modeling was used to discover that a loop connecting TMSs 10 and 11 in LacY probably moves during the transition between the unavailable outward‐open state and the outward‐occluded state. TMSs 11 and the other elements displaying a notable domain‐independent movement colocalize with the interdomain linker, suggesting that these elements could drive the alternating access movement between the domain halves. Preliminary evidence indicates that analogous movements occur in other members of the major facilitator superfamily. Proteins 2015; 83:735–745. © 2015 Wiley Periodicals, Inc.     

Latitudinal variation in resistance and tolerance to herbivory in the perennial herb Lythrum salicaria is related to intensity of herbivory and plant phenology

Both the length of the growing season and the intensity of herbivory often vary along climatic gradients, which may result in divergent selection on plant phenology, and on resistance and tolerance to herbivory. In Sweden, the length of the growing season and the number of insect herbivore species feeding on the perennial herb Lythrum salicaria decrease from south to north. Previous common‐garden experiments have shown that northern L. salicaria populations develop aboveground shoots earlier in the summer and finish growth before southern populations do. We tested the hypotheses that resistance and tolerance to damage vary with latitude in L. salicaria and are positively related to the intensity of herbivory in natural populations. We quantified resistance and tolerance of populations sampled along a latitudinal gradient by scoring damage from natural herbivores and fitness in a common‐garden experiment in the field and by documenting oviposition and feeding preference by specialist leaf beetles in a glasshouse experiment. Plant resistance decreased with latitude of origin, whereas plant tolerance increased. Oviposition and feeding preference in the glasshouse and leaf damage in the common‐garden experiment were negatively related to damage in the source populations. The latitudinal variation in resistance was thus consistent with reduced selection from herbivores towards the northern range margin of L. salicaria. Variation in tolerance may be related to differences in the timing of damage in relation to the seasonal pattern of plant growth, as northern genotypes have developed further than southern have when herbivores emerge in early summer.     

Mutant huntingtin causes metabolic imbalance by disruption of hypothalamic neurocircuits

In Huntington's disease (HD), the mutant huntingtin protein is ubiquitously expressed. The disease was considered to be limited to the basal ganglia, but recent studies have suggested a more widespread pathology involving hypothalamic dysfunction. Here we tested the hypothesis that expression of mutant huntingtin in the hypothalamus causes metabolic abnormalities. First, we showed that bacterial artificial chromosome-mediated transgenic HD (BACHD) mice developed impaired glucose metabolism and pronounced insulin and leptin resistance. Selective hypothalamic expression of a short fragment of mutant huntingtin using adeno-associated viral vectors was sufficient to recapitulate these metabolic disturbances. Finally, selective hypothalamic inactivation of the mutant gene prevented the development of the metabolic phenotype in BACHD mice. Our findings establish a causal link between mutant huntingtin expression in the hypothalamus and metabolic dysfunction, and indicate that metabolic parameters are powerful readouts to assess therapies aimed at correcting dysfunction in HD by silencing huntingtin expression in the brain.     

Mitogenome rearrangement in the cold-water scleractinian coral Lophelia pertusa (Cnidaria, Anthozoa) involves a long-term evolving group I intron

Group I introns are genetic insertion elements that invade host genomes in a wide range of organisms. In metazoans, however, group I introns are extremely rare, so far only identified within mitogenomes of hexacorals and some sponges. We sequenced the complete mitogenome of the cold-water scleractinian coral Lophelia pertusa, the dominating deep sea reef-building coral species in the North Atlantic Ocean. The mitogenome (16,150 bp) has the same gene content but organized in a unique gene order compared to that of other known scleractinian corals. A complex group I intron (6460 bp) inserted in the ND5 gene (position 717) was found to host seven essential mitochondrial protein genes and one ribosomal RNA gene. Phylogenetic analysis supports a vertical inheritance pattern of the ND5-717 intron among hexacoral mitogenomes with no examples of intron loss. Structural assessments of the Lophelia intron revealed an unusual organization that lacks the universally conserved ωG at the 3′ end, as well as a highly compact RNA core structure with overlapping ribozyme and protein coding capacities. Based on phylogenetic and structural analyses we reconstructed the evolutionary history of ND5-717, from its ancestral protist origin, through intron loss in some early metazoan lineages, and into a compulsory feature with functional implications in hexacorals.