Epitope mapping using mRNA display and a unidirectional nested deletion library

In vitro selection targeting an anti-polyhistidine monoclonal antibody was performed using mRNA display with a random, unconstrained 27-mer peptide library. After six rounds of selection, epitope-like peptides were identified that contain two to five consecutive, internal histidines and are biased for arginine residues, without any other identifiable consensus. The epitope was further refined by constructing a high-complexity, unidirectional fragment library from the final selection pool. Selection by mRNA display minimized the dominant peptide from the original selection to a 15-residue functional sequence (peptide Cmin: RHDAGDHHHHHGVRQ; K-D = 38 nM). Other peptides recovered from the fragment library selection revealed a separate consensus motif (ARRXA) C-terminal to the histidine track. Kinetics measurements made by surface plasmon resonance, using purified Fab (antigen-binding fragment) to prevent avidity effects, demonstrate that the selected peptides bind with 10- to 75-fold higher affinities than a hexahistidine peptide. The highest affinity peptides (K-D approximate to 10 nM) encode both a short histidine track and the ARRXA motif, suggesting that the motif and other flanking residues make important contacts adjacent to the core polyhistidine-binding site and can contribute > 2.5 kcal/mol of binding free energy. The fragment library construction methodology described here is applicable to the development of high-complexity protein or cDNA expression libraries for the identification of protein-protein interaction domains

Implications for operationalising the new education standards for nursing

Registrants and higher education institutions (HEIs) were recently invited to take part in the Nursing and Midwifery Council (NMC) consultation regarding its potential new standards for pre-registration nursing education. The consultation set out fresh standards for pre-registration nursing, together with an education framework that underpins the associated theory and practice. While these documents do not contain the final standards (NMC Council is set to approve them in spring 2018), they offer a glimpse of what the newly registered nurse should know and be able to do at the point of registration in order to practise safely and effectively and continue to develop their expertise. The draft proficiency standards are referred to in the document as being ‘ambitious in setting out the enhanced knowledge and skills that people can expect from nurses in the future’.It is interesting to look at the document in terms of practice learning, particularly in relation to by whom and how student nurses will be supervised and assessed in clinical practice and what the educational requirement should be. In this column, we offer some personal perspectives about the potential impact of these new standards in practice

Critical exploration of the new NMC future nurse : standards of proficiency for registered nurses

Introduction Following a two year process that included consulting and working alongside key stakeholder groups such as students, educators, healthcare professionals, charities and patient groups from across the UK, The Nursing and Midwifery Council (NMC) launched their Future nurse: Standards of proficiency for registered nurses (NMC 2018a). These new and compulsory standards are available for use in curricula by Approved Education Institutions (AEI’s) from the 28th January 2019. Only students who are part way through their pre-registration programme having commenced at an AEI prior to January 2019 will continue on the existing curriculum with the current NMC standards. The NMC refer to the standards as: “Ambitious new standards that set out the skills and knowledge the next generation of nurses will learn to enable them to deliver world class care” (NMC 2018b). The standards are set out in three parts: Part 1: Standards framework for nursing and midwifery education- this document provides a framework of five headings that underpin nurse education and training: Learning culture Educational governance and quality Student learning and empowerment Educators and assessors Curricula and assessment Part 2: Standards for student supervision and assessment- this document sets out the expectations for the learning, support, supervision and assessment (of theory and practice) of students in the practice environment. Part 3: Programme standards, which are the standards specific for each pre-registration or post-registration programme- this document set out the legal requirements for all pre-registration nursing education programmes. Collectively the three documents contain proficiencies that specify the knowledge and skills that registered nurses must demonstrate when caring for people of all ages and across all care settings, reflecting what the public can expect nurses to know and be able to do in order to deliver safe, compassionate and effective nursing care. A key message and something which remains consistent with the current pre-registration standards is the fundamental requirement for partnerships between Approved Education Institutions (AEIs) and healthcare organizations to provide the practice based learning for the student nurses: “(AEIs) are responsible for working with practice learning partners to manage the quality of their educational programmes. Overall responsibility for the day-to-day management of the quality of any educational programme lies with an AEI in partnership with practice learning partners who provide opportunities for practice experience to nursing and midwifery students” (NMC 2018a:4). This paper focuses on Part 2: standards for student support and assessment, exploring the key differences between the new and old sets of standards. We offer some personal perspectives about the potential impact of these new standards, particularly around the personnel responsible for supervising and assessing students in clinical practice and the potential preparation for the role. Partnership between AEIs and its practice partners are also explored

Characterisation of friction and lubrication regimes in premium tubular connections

A friction test rig has been developed to carry out repeated sliding friction tests for premium tubular connections. The test rig enables accurate measurement of friction in various contact regimes which are relevant to the threaded connections between tubular components. Higher load tests can simulate the contact in metal-to-metal seals under very high contact pressures by using perpendicular pin-on-pin tests. The contact in the thread loading flank under intermediate pressures can be simulated by using larger radius coupon-on-coupon tests. The measured coefficient of friction is well correlated with a lubrication parameter combining lubricant film thickness and initial surface roughness. © 2012 Elsevier Ltd. All rights reserved

Seasonal Mg/Ca-inferred temperatures of brackish water ostracods

The common brackish water ostracods Cyprideis torosa and Loxoconcha elliptica frequently occur together in high abundances in marginal marine environments. Seasonality of calcification differs between species and can have important implications for palaeotemperature reconstructions. There are existing palaeotemperature calibrations for both genera. However, the Loxoconcha spp. Mg/Ca temperature calibration has not thus far been applied to L. elliptica. The equation for Loxoconcha spp. does not rely on a known Mg/Cawater value, unlike the calibration for C. torosa, suggesting it may be possible to reconstruct temperatures without an estimation of Mg/Cawater, which is potentially particularly beneficial in environments that have highly dynamic Mg/Cawater. However, the calibration has only been applied in environments with marine-like Mg/Cawater. Demonstrating the applicability of the equation in marginal marine environments (with Mg/Cawater 3–5 mol/mol) and tracking the seasonal calcification of L. elliptica alongside C. torosa, therefore, has the potential to improve uncertainty in seasonal palaeotemperature reconstructions. Here, we compare previous monitoring of C. torosa with L. elliptica from the same collections. We demonstrate that the Mg/Ca temperature calibration for Loxoconcha spp. is appropriate to use with L. elliptica. Mg/Ca-inferred temperatures broadly track spring temperatures and suggest spring calcification. Cyprideis torosa Mg/Ca-inferred temperatures record the range of expected temperatures between spring and autumn. When analysing multiple single valves of L. elliptica and C. torosa simultaneously, the maximum Mg/CaC.torosa can, therefore, be used to reconstruct maximum summer temperatures, the minimum Mg/CaC.torosa to reconstruct autumn temperatures, and the Mg/CaL.elliptica to reconstruct the range in spring temperatures

A Peptide Core Motif for Binding to Heterotrimeric G Protein α Subunits

Recently, in vitro selection using mRNA display was used to identify a novel peptide sequence that binds with high affinity to G{alpha}i1. The peptide was minimized to a 9-residue sequence (R6A-1) that retains high affinity and specificity for the GDP-bound state of G{alpha}i1 and acts as a guanine nucleotide dissociation inhibitor (GDI). Here we demonstrate that the R6A-1 peptide interacts with G{alpha} subunits representing all four G protein classes, acting as a core motif for G{alpha} interaction. This contrasts with the consensus G protein regulatory(GPR) sequence, a 28-mer peptide GDI derived from the GoLoco (G{alpha}i/0-Loco interaction)/GPR motif that shares no homology with R6A-1 and binds only to G{alpha}i1-3 in this assay. Binding of R6A-1 is generally specific to the GDP-bound state of the G{alpha} subunits and excludes association with G{beta}{gamma}. R6A-G{alpha}i1 complexes are resistant to trypsin digestion and exhibit distinct stability in the presence of Mg2+, suggesting that the R6A and GPR peptides exert their activities using different mechanisms. Studies using G{alpha}i1/G{alpha}s chimeras identify two regions of G{alpha}i1 (residues 1–35 and 57–88) as determinants for strong R6A-Gi{alpha}1 interaction. Residues flanking the R6A-1 peptide confer unique binding properties, indicating that the core motif could be used as a starting point for the development of peptides exhibiting novel activities and/or specificity for particular G protein subclasses or nucleotide-bound states

Virtual outreach: economic evaluation of joint teleconsultations for patients referred by their general practitioner for a specialist opinion

Objectives To test the hypotheses that, compared with conventional outpatient consultations, joint teleconsultation (virtual outreach) would incur no increased costs to the NHS, reduce costs to patients, and reduce absences from work by patients and their carers.Design Cost consequences study alongside randomised controlled trial.Setting Two hospitals in London and Shrewsbury and 29 general practices in inner London and Wales.Participants 3170 patients identified; 2094 eligible for inclusion and willing to participate. 1051 randomised to virtual outreach and 1043 to standard outpatient appointments.Main outcome measures NHS costs, patient costs, health status (SF-12), time spent attending index consultation, patient satisfaction.Results Overall six month costs were greater for the virtual outreach consultations (pound724 per patient) than for conventional outpatient appointments (pound625): difference in means pound99 ($162; is not an element of138) (95% confidence interval pound10 to pound187, P=0.03). if the analysis is restricted to resource items deemed "attributable" to the index consultation, six month costs were still greater for virtual outreach: difference in means pound108 (pound73 to pound142, P < 0.0001). In both analyses the index consultation accounted for the excess cost. Savings to patients in terms of costs and time occurred in both centres: difference in mean total patient cost 8 pound (5 pound to 10 pound, P < 0.0001). Loss of productive time was less in the virtual outreach group: difference in mean cost pound11 (pound10 to pound12, P < 0.0001).Condusion The main hypothesis that virtual outreach would be cost neutral is rejected, but the hypotheses that costs to patients and losses in productivity would be lower are supported

Laryngeal Nerve Activity During Pulse Emission in the CF-FM Bat, Rhinolophus ferrumequinum. I. Superior Laryngeal Nerve (External Motor Branch)

The activity of the external (motor) branch of the superior laryngeal nerve (SLN), innervating the cricothyroid muscle, was recorded in the greater horseshoe bat,Rhinolophus ferrumequinum. The bats were induced to change the frequency of the constant frequency (CF) component of their echolocation signals by presenting artificial signals for which they Doppler shift compensated. The data show that the SLN discharge rate and the frequency of the emitted CF are correlated in a linear manner