GPCR structure, function, drug discovery and crystallography: report from Academia-Industry International Conference (UK Royal Society) Chicheley Hall, 1-2 September 2014

G-protein coupled receptors (GPCRs) are the targets of over half of all prescribed drugs today. The UniProt database has records for about 800 proteins classified as GPCRs, but drugs have only been developed against 50 of these. Thus, there is huge potential in terms of the number of targets for new therapies to be designed. Several breakthroughs in GPCRs biased pharmacology, structural biology, modelling and scoring have resulted in a resurgence of interest in GPCRs as drug targets. Therefore, an international conference, sponsored by the Royal Society, with world-renowned researchers from industry and academia was recently held to discuss recent progress and highlight key areas of future research needed to accelerate GPCR drug discovery. Several key points emerged. Firstly, structures for all three major classes of GPCRs have now been solved and there is increasing coverage across the GPCR phylogenetic tree. This is likely to be substantially enhanced with data from x-ray free electron sources as they move beyond proof of concept. Secondly, the concept of biased signalling or functional selectivity is likely to be prevalent in many GPCRs, and this presents exciting new opportunities for selectivity and the control of side effects, especially when combined with increasing data regarding allosteric modulation. Thirdly, there will almost certainly be some GPCRs that will remain difficult targets because they exhibit complex ligand dependencies and have many metastable states rendering them difficult to resolve by crystallographic methods. Subtle effects within the packing of the transmembrane helices are likely to mask and contribute to this aspect, which may play a role in species dependent behaviour. This is particularly important because it has ramifications for how we interpret pre-clinical data. In summary, collaborative efforts between industry and academia have delivered significant progress in terms of structure and understanding of GPCRs and will be essential for resolving problems associated with the more difficult targets in the future.

Assisstantship improves medical student preparedness for starting work

Background: The GMC has recommended introducing student assistantships during which final year students, under supervision, undertake most of the responsibilities of a FY1 doctor. The Medical School at Queen’s University Belfast in 2011/12 introduced an assistantship programme. We have evaluated the impact of the assistantship on students’ perception of their preparedness for starting work.
Methods: Students were asked to complete a questionnaire at the beginning of the assistantship. It assessed the students’ perception of their preparedness in five areas: clinical and practical skills, communications skills, teaching and learning, understanding the work environment and team working. After the assistantship they again completed the questionnaire. Comparison of the results allowed an assessment of the impact of the assistantship.
Results: There was a statistically significant improvement in the students' perception of their preparation for 49 of 56 tasks contained within the questionnaire. After the assistantship 81.2% of students felt well prepared for starting work compared with 38.9% before the assistantship. 93.9% agreed that the assistantship had improved their preparedness for starting work.
Conclusions: The assistantship at Queen’s University improves medical students’ perception of their preparedness for starting work. The majority of medical students feel well prepared for starting work after completing the assistantship.

Conduit artery structure and function in lowlanders and native highlanders: relationships with oxidative stress and role of sympathoexcitation

Research detailing the normal vascular adaptions to high altitude is minimal and often confounded by pathology (e.g. chronic mountain sickness) and methodological issues. We examined vascular function and structure in: (1) healthy lowlanders during acute hypoxia and prolonged (∼2 weeks) exposure to high altitude, and (2) high-altitude natives at 5050 m (highlanders). In 12 healthy lowlanders (aged 32 ± 7 years) and 12 highlanders (Sherpa; 33 ± 14 years) we assessed brachial endothelium-dependent flow-mediated dilatation (FMD), endothelium-independent dilatation (via glyceryl trinitrate; GTN), common carotid intima–media thickness (CIMT) and diameter (ultrasound), and arterial stiffness via pulse wave velocity (PWV; applanation tonometry). Cephalic venous biomarkers of free radical-mediated lipid peroxidation (lipid hydroperoxides, LOOH), nitrite (NO₂^–) and lipid soluble antioxidants were also obtained at rest. In lowlanders, measurements were performed at sea level (334 m) and between days 3–4 (acute high altitude) and 12–14 (chronic high altitude) following arrival to 5050 m. Highlanders were assessed once at 5050 m. Compared with sea level, acute high altitude reduced lowlanders’ FMD (7.9 ± 0.4 vs. 6.8 ± 0.4%; P = 0.004) and GTN-induced dilatation (16.6 ± 0.9 vs. 14.5 ± 0.8%; P = 0.006), and raised central PWV (6.0 ± 0.2vs. 6.6 ± 0.3 m s⁻¹; P = 0.001). These changes persisted at days 12–14, and after allometrically scaling FMD to adjust for altered baseline diameter. Compared to lowlanders at sea level and high altitude, highlanders had a lower carotid wall:lumen ratio (∼19%, P ≤ 0.04), attributable to a narrower CIMT and wider lumen. Although both LOOH and NO₂^– increased with high altitude in lowlanders, only LOOH correlated with the reduction in GTN-induced dilatation evident during acute (n = 11, r = −0.53) and chronic (n = 7, r = −0.69; P ≤ 0.01) exposure to 5050 m. In a follow-up, placebo-controlled experiment (n = 11 healthy lowlanders) conducted in a normobaric hypoxic chamber (inspired O₂ fraction () = 0.11; 6 h), a sustained reduction in FMD was evident within 1 h of hypoxic exposure when compared to normoxic baseline (5.7 ± 1.6 vs. 8.0 ±1.3%; P < 0.01); this decline in FMD was largely reversed following α₁-adrenoreceptor blockade. In conclusion, high-altitude exposure in lowlanders caused persistent impairment in vascular function, which was mediated partially via oxidative stress and sympathoexcitation. Although a lifetime of high-altitude exposure neither intensifies nor attenuates the impairments seen with short-term exposure, chronic high-altitude exposure appears to be associated with arterial remodelling.

The scattered trans-Neptunian object 1998 XY95

On 1998 December 12 a new trans-Neptunian object, 1998 XY95, was discovered as part of a deep search. Recent observations of this object have placed it amongst the class of objects known as the scattered trans-Neptunian objects (TNOs). A total of 39 CCD images of 1998 XY95 were taken over two nights, and these were used to search for a light curve, but no significant periodicity was found. An examination of the possible orbital evolution gives no indication of how it may have arrived on its present orbit. The current best-fitting orbit is unstable, but remains within a band of semi-major axis approximately 2au wide. The bottom of this band is due to 3:1 mean motion resonance with Neptune, while the reason for the top of the band remains unclear.

Inland shell midden site-formation:Investigation into a late Pleistocene to early Holocene midden from Trang An, Northern Vietnam

Over the course of the past two decades there has been growing research interest in the site formation processes of shell middens. This stands along-side and is being used to inform cultural, dietary and palaeo-environmental reconstructions. Just as midden site formation processes have turned out to be many and varied, however, the kinds of shell-bearing sites that past human communities created are likely to have been no less diverse. Subsuming such sites under a single category - shell middens - normalises that variation and may lead to the misinterpretation of site function. The greater part of research in this field also continues to focus on coastal shell middens; comparatively little attention has been paid to middens containing freshwater and especially terrestrial molluscs from hinterland locations. As a result, much of the current understanding about shell-midden sites carries a spatial as well as a functional bias. This paper hopes to contribute towards discussion on both fronts. It presents a detailed examination of the formation processes that went into the creation of a land snail-dominated late- to post-glacial midden from northern Vietnam, and considers the role that it may have played in the early settlement of this area. The data presented comes from ongoing archaeological excavations at Hang Boi, a cave located in the sub-coastal karstic uplands of Trang An park, in the Vietnamese Province of Ninh Binh. (C) 2010 Elsevier Ltd and INQUA. All rights reserved.

Collating the knowledge base for core outcome set development: developing and appraising the search strategy for a systematic review

Background: The COMET (Core Outcome Measures in Effectiveness Trials) Initiative is developing a publicly accessible online resource to collate the knowledge base for core outcome set development (COS) and the applied work from different health conditions. Ensuring that the database is as comprehensive as possible and keeping it up to date are key to its value for users. This requires the development and application of an optimal, multi-faceted search strategy to identify relevant material. This paper describes the challenges of designing and implementing such a search, outlining the development of the search strategy for studies of COS development, and, in turn, the process for establishing a database of COS.

Methods: We investigated the performance characteristics of this strategy including sensitivity, precision and numbers needed to read. We compared the contribution of databases towards identifying included studies to identify the best combination of methods to retrieve all included studies.

Results: Recall of the search strategies ranged from 4% to 87%, and precision from 0.77% to 1.13%. MEDLINE performed best in terms of recall, retrieving 216 (87%) of the 250 included records, followed by Scopus (44%). The Cochrane Methodology Register found just 4% of the included records. MEDLINE was also the database with the highest precision. The number needed to read varied between 89 (MEDLINE) and 130 (SCOPUS).

Conclusions: We found that two databases and hand searching were required to locate all of the studies in this review. MEDLINE alone retrieved 87% of the included studies, but actually 97% of the included studies were indexed on MEDLINE. The Cochrane Methodology Register did not contribute any records that were not found in the other databases, and will not be included in our future searches to identify studies developing COS. SCOPUS had the lowest precision rate (0.77) and highest number needed to read (130). In future COMET searches for COS a balance needs to be struck between the work involved in screening large numbers of records, the frequency of the searching and the likelihood that eligible studies will be identified by means other than the database searches.

A randomised comparison of daunorubicin 90mg/m2 vs 60mg/m2 in AML induction: results from the UK NCRI AML17 trial in 1206 patients

Modifying induction therapy in AML may improve the remission rate and reduce the risk of relapse thereby improving survival. Escalation of the daunorubicin dose to 90mg/m(2) has shown benefit for some patient subgroups when compared with a dose of 45mg/m(2) and has been recommended as a standard of care. However 60mg/m(2) is widely used and has never been directly compared to 90mg/m(2). As part of the UK NCRI AML17 trial 1206 adults with untreated AML or high risk MDS, mostly under 60 years of age, were randomised to a first induction course of chemotherapy which delivered either 90mg/m(2) or 60mg/m(2) on days 1,3 and 5 combined with cytosine arabinoside. All patients then received a second course which included daunorubicin 50mg/m(2) on days 1,3 and 5. There was no overall difference in complete remission rate (CR) (73% vs 75%, OR1.07 (0.83-1.39), p=0.6) or in any recognised subgroup. The 60 day mortality was increased in the 90mg/m2 arm (10% vs 5% (HR 1.98(1.30-3.02) p=0.001)), which resulted in no difference in overall 2 year survival (59% vs 60%, HR 1.16(0.95-1.43), p=0.15). In exploratory subgroup analysis there was no subgroup which showed significant benefit, although there was a significant interaction by FLT3 ITD mutation. The trial is registered to www.isrctn.com as ISRCTN55675535.