Crystal structure of the µ-opioid receptor bound to a morphinan antagonist

Opium is one of the world's oldest drugs, and its derivatives morphine and codeine are among the most used clinical drugs to relieve severe pain. These prototypical opioids produce analgesia as well as many undesirable side effects (sedation, apnoea and dependence) by binding to and activating the G-protein-coupled μ-opioid receptor (μ-OR) in the central nervous system. Here we describe the 2.8?? crystal structure of the mouse μ-OR in complex with an irreversible morphinan antagonist. Compared to the buried binding pocket observed in most G-protein-coupled receptors published so far, the morphinan ligand binds deeply within a large solvent-exposed pocket. Of particular interest, the μ-OR crystallizes as a two-fold symmetrical dimer through a four-helix bundle motif formed by transmembrane segments 5 and 6. These high-resolution insights into opioid receptor structure will enable the application of structure-based approaches to develop better drugs for the management of pain and addiction.     

Reductive carboxylation supports growth in tumour cells with defective mitochondria

Mitochondrial metabolism provides precursors to build macromolecules in growing cancer cells. In normally functioning tumour cell mitochondria, oxidative metabolism of glucose- and glutamine-derived carbon produces citrate and acetyl-coenzyme A for lipid synthesis, which is required for tumorigenesis. Yet some tumours harbour mutations in the citric acid cycle (CAC) or electron transport chain (ETC) that disable normal oxidative mitochondrial function, and it is unknown how cells from such tumours generate precursors for macromolecular synthesis. Here we show that tumour cells with defective mitochondria use glutamine-dependent reductive carboxylation rather than oxidative metabolism as the major pathway of citrate formation. This pathway uses mitochondrial and cytosolic isoforms of NADP(+)/NADPH-dependent isocitrate dehydrogenase, and subsequent metabolism of glutamine-derived citrate provides both the acetyl-coenzyme A for lipid synthesis and the four-carbon intermediates needed to produce the remaining CAC metabolites and related macromolecular precursors. This reductive, glutamine-dependent pathway is the dominant mode of metabolism in rapidly growing malignant cells containing mutations in complex I or complex III of the ETC, in patient-derived renal carcinoma cells with mutations in fumarate hydratase, and in cells with normal mitochondria subjected to acute pharmacological ETC inhibition. Our findings reveal the novel induction of a versatile glutamine-dependent pathway that reverses many of the reactions of the canonical CAC, supports tumour cell growth, and explains how cells generate pools of CAC intermediates in the face of impaired mitochondrial metabolism.     

Whole-genome analysis informs breast cancer response to aromatase inhibition

To correlate the variable clinical features of oestrogen-receptor-positive breast cancer with somatic alterations, we studied pretreatment tumour biopsies accrued from patients in two studies of neoadjuvant aromatase inhibitor therapy by massively parallel sequencing and analysis. Eighteen significantly mutated genes were identified, including five genes (RUNX1, CBFB, MYH9, MLL3 and SF3B1) previously linked to haematopoietic disorders. Mutant MAP3K1 was associated with luminal A status, low-grade histology and low proliferation rates, whereas mutant TP53 was associated with the opposite pattern. Moreover, mutant GATA3 correlated with suppression of proliferation upon aromatase inhibitor treatment. Pathway analysis demonstrated that mutations in MAP2K4, a MAP3K1 substrate, produced similar perturbations as MAP3K1 loss. Distinct phenotypes in oestrogen-receptor-positive breast cancer are associated with specific patterns of somatic mutations that map into cellular pathways linked to tumour biology, but most recurrent mutations are relatively infrequent. Prospective clinical trials based on these findings will require comprehensive genome sequencing.     

Modeling an Elastic-Demand Bimodal Transport Network with Park-and-Ride Trips

This paper presents a network equilibrium formulation for modeling commuters＇ travel choices in a bimodal transport system with park-and-ride （P＆R） trips while the total demand is elastic to the congestion level of the network. A super-network approach is adopted in the proposed model. It is assumed that commuters＇ trips are categorized into two types, auto mode only and a combined mode with both auto and transit modes. The former is referred to as the pure mode trip and the latter as the P＆R mode trip. The proposed model simultaneously considers the commuter＇s choice of the pure mode versus the P＆R mode, the choice of parking location for the pure mode, the choice of transfer point for the P＆R mode, as well as the route choice for each mode. The demand elasticity of transport system, the capacity constraints of transport facilities, and the congestion interaction throughout the super-network are also explicitly incorporated into the proposed model. The results of the numerical experiment show the following key findings： （i） traditional parking/P＆R models may overestimate or underestimate travel demand distribution over network; （ii） parking/P＆R, transit scheduling, and carpooling schemes bring significant impacts on commuters＇ travel behavior and network performance; and （iii） different transport policies may be to some extent mutually substituted .     

Reproductive biology, hybridization and flower visitors of rare Sclerocactus taxa in Utah's Uintah Basin

We studied the mating system and flower visitors of 2 threatened species of Sclerocactus (Cactaceae) in the Uintah Basin of eastern Utah—an area undergoing rapid energy development. We found that both S. wetlandicus and S. brevispinus are predominantly outcrossed and are essentially self-incompatible. A third presumptive taxon (undescribed; here called S. wetlandicus -var1) is fully self-compatible but cannot produce seeds unless the flowers are visited by pollinators. We found tentative evidence for pollinator limitation of fruit set in S. brevispinus but not in the other taxa. Pollinators are largely species of native ground-nesting bees in the subfamily Halictinae. These bees are indiscriminate collectors of pollen and nectar from Sclerocactus flowers and other plant species. The well-being of Sclerocactus populations requires that land managers ensure the preservation of halictid bee habitat, which includes both nesting sites and a diversity of floral resources. These bees are also undoubtedly the pollen vectors responsible for the production of hybrids in areas where S. wetlandicus and S. brevispinus meet. We experimentally confirmed that reciprocal crosses between S. wetlandicus and S. brevispinus are indistinguishable from each other or from pure crosses in fruits/flower, seeds/fruit and seed weights. We suggest that hybridization preceded energy development, that it attests to the ongoing process of evolution in the Sclerocactus group, and that it should not be suppressed by anthropogenic activity.     

Leafhopper assemblages on native and reseeded grasslands in southwestern Montana

Using sweep samples, we surveyed leafhoppers (Homoptera: Cicadellidae) on grassland sites in the Gallatin Valley of Montana during 1988 and 1991. We sampled 12 sites representing 2 habitat types defined by their dominant plant species in an undisturbed state ( Stipa comata / Bouteloua gracilis and Festuca idahoensis / Agropyron spicatum ). At half of the sites the native plant communities were present, whereas the remainder had been reseeded with either Agropyron spicatum (to replace the S. comata / B. gracilis assemblage ) or Bromus inermis (to replace the F. idahoensis / A. spicatum assemblage). We found at least 66 species of leafhoppers among 44,428 adults collected. Seven taxa comprised 83% of all individuals collected: Doratura stylata (26%), Ceratagallia spp. (18%), Endria inimica (17%), Orocastus perpusillus (7%), Sorhoanus spp. (6%), Athysanella spp. (5%), and Psammotettix lividellus (4%). Sites with similar vegetation had broadly similar leafhopper assemblages, and assemblages differed most between the relatively xeric Stipa comata / Bouteloua gracilis sites and the more mesic sites dominated by Bromus inermis . The composition of a leafhopper assemblage at a site tended to be more similar to those on noncontiguous sites with the same overall vegetation than to those on contiguous sites with different vegetation. These patterns are likely related to the fact that many Cicadellidae are host specialists. In fact, variation in abundance of some of the most common leafhopper taxa on our sites was correlated with the percent cover of their known host plants. Our analyses of the leafhopper assemblages generally support the contention that terrestrial plant associations are among the more useful indicators of insect community composition.     

Range extension and ecological information for Orconectes virilis (Hagen 1870) (Decapoda: Cambaridae) in Idaho, USA

We report the 1st record of the crayfish Orconectes virilis (Hagen 1870) from Idaho and the Pacific Northwest, USA. We also provide an updated checklist of recent Idaho crayfishes, which now includes 2 families (Astacidae and Cambaridae), 3 genera ( Pacifasticus, Orconectes , and Procambarus ), and 7 species and subspecies ( Pacifasticus connectus (Faxon 1914), P. gambelii (Girard 1852), P. leniusculus leniusculus (Dana 1852), P. leniusculus klamathensis (Stimpson 1857), P. leniusculus trowbridgii (Stimpson 1857), O. virilis , and Procambarus clarkii (Girard 1852). Native crayfish were not found at the O. virilis collection sites. The localities where O. virilis was found were characterized by low-gradient streams impacted by fine sediment. Aquatic invertebrates found in association with O. virilis tended to be the more pollution-tolerant taxa.