Meeting the UK Government's prevention agenda: primary care practitioners can be trained in skills to prevent disease and support self-management.

AIMS: The NHS Long Term Plan has a prevention focus and ambition to support patients to self-manage disease through improving health behaviours. An essential requirement of self-management is behaviour change, but many practitioners have not been trained in skills to support behaviour change. 'Healthy Conversation Skills' (HCS) training was developed at the University of Southampton for this purpose. This article reports on a pilot study that aimed to assess the feasibility of primary care practitioners adopting HCS in their routine practice. It describes their experiences and level of competence post-training. METHODS: Health Education England (Wessex) commissioned HCS training for 18 primary care practitioners. Fifteen of these practitioners were subsequently observed in their consultations at one or two time points; face-to-face semi-structured, reflective feedback interviews were conducted immediately following the observations. Practitioners' HCS competence was assessed from the observations and interviews using a previously developed and published coding rubric. The interview data were analysed thematically to understand practitioners' experiences of using the new skills. RESULTS: Practitioners demonstrated competence in embedding the skills into their routine practice following HCS training. They reflected on how patients liked being asked questions, the usefulness of setting SMARTER (Specific, Measured, Action-oriented, Realistic, Timed, Evaluated and Reviewed) goals and the power of listening. They could also identify facilitators of skill use and ways to overcome challenges such as patients with competing priorities and organisational constraints. They found the skills valuable as a way of empowering patients to make changes to manage their own health. CONCLUSIONS: HCS are acceptable to primary care practitioners, can be readily adopted into their routine consultations and are a helpful strategy for supporting patients to make changes. HCS training has the potential to be a sustainable, scalable and effective way of contributing to the prevention agenda by supporting disease self-management, and hence of addressing today's epidemic of lifestyle-related conditions.

Modulating medial prefrontal cortex activity using real-time fMRI neurofeedback: Effects on reality monitoring performance and associated functional connectivity.

Neuroimaging studies have found 'reality monitoring', our ability to distinguish internally generated experiences from those derived from the external world, to be associated with activity in the medial prefrontal cortex (mPFC) of the brain. Here we probe the functional underpinning of this ability using real-time fMRI neurofeedback to investigate the involvement of mPFC in recollection of the source of self-generated information. Thirty-nine healthy individuals underwent neurofeedback training in a between groups study receiving either Active feedback derived from the paracingulate region of the mPFC (21 subjects) or Sham feedback based on a similar level of randomised signal (18 subjects). Compared to those in the Sham group, participants receiving Active signal showed increased mPFC activity over the course of three real-time neurofeedback training runs undertaken in a single scanning session. Analysis of resting state functional connectivity associated with changes in reality monitoring accuracy following Active neurofeedback revealed increased connectivity between dorsolateral frontal regions of the fronto-parietal network (FPN) and the mPFC region of the default mode network (DMN), together with reduced connectivity within ventral regions of the FPN itself. However, only a trend effect was observed in the interaction of the recollection of the source of Imagined information compared with recognition memory between participants receiving Active and Sham neurofeedback, pre- and post- scanning. As such, these findings demonstrate that neurofeedback can be used to modulate mPFC activity and increase cooperation between the FPN and DMN, but the effects on reality monitoring performance are less clear.

The use of intravenous lidocaine for postoperative pain and recovery: international consensus statement on efficacy and safety.

Intravenous lidocaine is used widely for its effect on postoperative pain and recovery but it can be, and has been, fatal when used inappropriately and incorrectly. The risk-benefit ratio of i.v. lidocaine varies with type of surgery and with patient factors such as comorbidity (including pre-existing chronic pain). This consensus statement aims to address three questions. First, does i.v. lidocaine effectively reduce postoperative pain and facilitate recovery? Second, is i.v. lidocaine safe? Third, does the fact that i.v. lidocaine is not licensed for this indication affect its use? We suggest that i.v. lidocaine should be regarded as a 'high-risk' medicine. Individual anaesthetists may feel that, in selected patients, i.v. lidocaine may be beneficial as part of a multimodal peri-operative pain management strategy. This approach should be approved by hospital medication governance systems, and the individual clinical decision should be made with properly informed consent from the patient concerned. If i.v. lidocaine is used, we recommend an initial dose of no more than 1.5 mg.kg-1 , calculated using the patient's ideal body weight and given as an infusion over 10 min. Thereafter, an infusion of no more than 1.5 mg.kg-1 .h-1 for no longer than 24 h is recommended, subject to review and re-assessment. Intravenous lidocaine should not be used at the same time as, or within the period of action of, other local anaesthetic interventions. This includes not starting i.v. lidocaine within 4 h after any nerve block, and not performing any nerve block until 4 h after discontinuing an i.v. lidocaine infusion.

The use of ultrasound to assess fetal growth in a guinea pig model of fetal growth restriction.

Fetal growth restriction (FGR) is a common and potentially severe pregnancy complication. Currently there is no treatment available. The guinea pig is an attractive model of human pregnancy as placentation is morphologically very similar between the species. Nutrient restriction of the dam creates growth-restricted fetuses while leaving an intact uteroplacental circulation, vital for evaluating novel therapies for FGR. Growth-restricted fetuses were generated by feeding Dunkin Hartley guinea pig dams 70% of ad libitum intake from four weeks before and throughout pregnancy. The effect of maternal nutrient restriction (MNR) on dams and fetuses was carefully monitored, and ultrasound measurements of pups collected. There was no difference in maternal weight at conception, however by five weeks post conception MNR dams were significantly lighter ( P < 0.05). MNR resulted in significantly smaller pup size from 0.6-0.66 gestation. Ultrasound is a powerful non-invasive tool for assessing the effect of therapeutic interventions on fetal growth, allowing longitudinal measurement of fetuses. This model and method yield data applicable to the human condition without the need for animal sacrifice and will be useful in the translation of therapies for FGR into the clinic.

Time to get comfortable with a labour epidural.

Women frequently request regional analgesia during labour, yet little is known about how long it takes before they become comfortable. This prospective observational study aimed to determine various time-points following maternal request for regional analgesia in labour until comfort was achieved. It was conducted in two tertiary referral centres for maternity care in Australia between December 2009 and May 2010.Midwives and anaesthetists recorded times of maternal request for regional analgesia, anaesthetist contact,anaesthetist's arrival in the labour room, local anaesthetic infiltration on starting the procedure, injection of neuraxial local anaesthetic and first report of maternal comfort. Composite median times and interquartile range were recorded for maternal request to anaesthetist arrival, anaesthetist arrival to maternal comfort and total time from request to comfort. Statistical modelling and regression analyses assessed possible factors associated with these time intervals. A P value <0.05 was considered significant. Of the 324 maternal requests, 244 out of 324 (75.3%, 95% confidence interval 70.2% to 79.9%) were recorded as having achieved satisfactory labour analgesia. Median interquartile range times observed were: maternal request to anaesthetist arrival: 20 (10 to 35) minutes; anaesthetist arrival to maternal comfort: 40 (30 to 50) minutes; and total time from request to comfort: 65 (50 to 85) minutes. We have shown that approximately one hour is required for a mother to achieve comfort following her request for epidural analgesia during labour. Our findings are likely to provide useful information for antenatal education, enhance informed consent and improve the provision of anaesthetic services for labour analgesia.

Structural basis of the interaction of the breast cancer oncogene LMO4 with the tumour suppressor CtIP/RBBP8.

LIM-only protein 4 (LMO4) is strongly linked to the progression of breast cancer. Although the mechanisms underlying this phenomenon are not well understood, a role is emerging for LMO4 in regulation of the cell cycle. We determined the solution structure of LMO4 in complex with CtIP (C-terminal binding protein interacting protein)/RBBP8, a tumour suppressor protein that is involved in cell cycle progression, DNA repair and transcriptional regulation. Our data reveal that CtIP and the essential LMO cofactor LDB1 (LIM-domain binding protein 1) bind to the same face on LMO4 and cannot simultaneously bind to LMO4. We hypothesise that overexpression of LMO4 may disrupt some of the normal tumour suppressor activities of CtIP, thereby contributing to breast cancer progression.

Computation of Solar Radiative Fluxes by 1D and 3D Methods Using Cloudy Atmospheres Inferred from A-train Satellite Data.

This study used realistic representations of cloudy atmospheres to assess errors in solar flux estimates associated with 1D radiative transfer models. A scene construction algorithm, developed for the EarthCARE mission, was applied to CloudSat, CALIPSO and MODIS satellite data thus producing 3D cloudy atmospheres measuring 61 km wide by 14,000 km long at 1 km grid-spacing. Broadband solar fluxes and radiances were then computed by a Monte Carlo photon transfer model run in both full 3D and 1D independent column approximation modes. Results were averaged into 1,303 (50 km)2 domains. For domains with total cloud fractions Ac  < 0.7 top-of-atmosphere (TOA) albedos tend to be largest for 3D transfer with differences increasing with solar zenith angle. Differences are largest for Ac  > 0.7 and characterized by small bias yet large random errors. Regardless of Ac , differences between 3D and 1D transfer rarely exceed ±30 W m-2 for net TOA and surface fluxes and ±10 W m-2 for atmospheric absorption. Horizontal fluxes through domain sides depend on Ac with ∼20% of cases exceeding ±30 W m-2; the largest values occur for Ac  > 0.7. Conversely, heating rate differences rarely exceed ±20%. As a cursory test of TOA radiative closure, fluxes produced by the 3D model were averaged up to (20 km)2 and compared to values measured by CERES. While relatively little attention was paid to optical properties of ice crystals and surfaces, and aerosols were neglected entirely, ∼30% of the differences between 3D model estimates and measurements fall within ±10 W m-2; this is the target agreement set for EarthCARE. This, coupled with the aforementioned comparison between 3D and 1D transfer, leads to the recommendation that EarthCARE employ a 3D transport model when attempting TOA radiative closure.

Development of a simple extraction and clean-up procedure for determination of organochlorine pesticides in soil using gas chromatography-tandem mass spectrometry.

A procedure based on QuEChERS extraction and a simultaneous liquid-liquid partition clean-up was developed. The procedure involved extraction of hydrated soil samples using acetonitrile and clean-up by liquid-liquid partition into n-hexane. The hexane extracts produced were clean and suitable for determination using gas chromatography-tandem mass spectrometry (GC-MS/MS). The method was validated by analysis of soil samples, spiked at five levels between 1 and 200 microg kg(-1). The recovery values were generally between 70 and 100% and the relative standard deviation values (%RSDs) were at or below 20%. The procedure was validated for determination of 19 organochlorine (OC) pesticides. These were hexachlorobenzene (HCB), alpha-HCH, beta-HCH, gamma-HCH, heptachlor, heptachlor epoxide (trans), aldrin, dieldrin, chlordane (trans), chlordane (cis), oxychlordane, alpha-endosulfan, beta-endosulfan, endosulfan sulfate, endrin, p,p'-DDT, o,p'-DDT, p,p'-DDD and p,p'-DDE. The method achieved low limits of detection (LOD; typically 0.3 microg kg(-1)) and low limits of quantification (LOQ; typically 1.0 microg kg(-1)). The method performance was also assessed using five fortified soil samples with different physico-chemical properties and the method performance was consistent for the different types of soil samples. The proposed method was compared with an established procedure based on Soxtec extraction. This comparison was carried out using six soil samples collected from regions of Pakistan with a history of intensive pesticide use. The results of this comparison showed that the two procedures produced results with good agreement. The proposed method produced cleaner extracts and therefore led to lower limits of quantification. The proposed method was less time consuming and safer to use. The six samples tested during this comparison showed that soils from cotton growing regions contained a number of persistent OC residues at relatively low levels (<10 microg kg(-1)). These residues were alpha-HCH, gamma-HCH, heptachlor, chlordane (trans), p,p'-DDT, o,p'-DDT, p,p'-DDD, p,p'-DDE, beta-endosulfan and endosulfan sulfate.

Cystic fibrosis neutrophils have normal intrinsic reactive oxygen species generation.

Previous studies have identified abnormalities in the oxidative responses of the neutrophil in cystic fibrosis (CF), but it is unclear whether such changes relate to loss of membrane cystic fibrosis transmembrane conductance regulator (CFTR) or to the inflammatory environment present in this disease. The aim of the present study was to determine whether neutrophils from CF patients demonstrate an intrinsic abnormality of the respiratory burst. The respiratory burst activity of neutrophils isolated from stable DeltaF508 homozygote CF patients and matched healthy controls was quantified by both chemiluminscence and cytochrome C reduction. Expression of NADPH oxidase components and CFTR was determined by Western blotting and RT-PCR. The oxidative output from neutrophils from CF in response to receptor-linked and particulate stimuli did not differ from that of controls. Expression of NADPH oxidase components was identical in CF and non-CF neutrophils. While low levels of CFTR mRNA could be identified in the normal human neutrophil, we were unable to detect CFTR protein in human neutrophil lysates or immunoprecipitates. CFTR has no role in controlling neutrophil oxidative activity; previously reported differences in neutrophil function between CF and non-CF subjects most likely relate to the inflammatory milieu from which the cells were isolated.

Clostridium difficile pancolitis in adults with cystic fibrosis.

We report three cases of Clostridium difficile pancolitis in adults with cystic fibrosis (CF) in whom the presenting symptoms were atypical. All three required treatment with systemic steroids, in addition to oral vancomycin and metronidazole to achieve resolution of the colitis. This experience suggests that C. difficile colitis should be considered in individuals with CF presenting with non-specific abdominal symptoms.

A new platelet parameter, the mean platelet component, can demonstrate abnormal platelet function and structure in myelodysplasia.

The mean platelet component (MPC) is a new platelet parameter generated by the Bayer ADVIA 120 full blood count analyser as part of the routine complete blood count (CBC) test cycle. We report a case of myelodysplasia with bleeding complications and abnormal template bleeding time in whom low mean platelet component parameters were associated with partial platelet granule deficiency, demonstrated by transmission electron microscopy. We suggest that the mean platelet component is an inexpensive and rapid test to screen for platelet dysfunction related to ultrastructural abnormalities in myelodysplasia.

Umbravirus gene expression helps potato leafroll virus to invade mesophyll tissues and to be transmitted mechanically between plants.

Potato leafroll virus (PLRV) was mechanically transmissible when inocula also contained the umbravirus Pea enation mosaic virus-2 (PEMV-2). In plants infected with PLRV and PEMV-2, PLRV accumulated in clusters of mesophyll cells in both inoculated and systemically infected leaves. No transmissions were obtained by coinoculation with Potato virus Y, Potato virus X (PVX), Tobacco mosaic virus, or Cucumber mosaic virus (CMV), although PLRV was transmissible from mixtures with CMV(ORF4) (a recombinant that contained the movement protein (MP) gene of the umbravirus Groundnut rosette virus (GRV) in place of the CMV MP gene). In contrast, neither a recombinant PVX that expressed GRV MP nor a mutant of CMV(ORF4), in which the CMV 2b gene was untranslatable, was able to help PLRV transmission. Possibly both a cell-to-cell movement function and counterdefense mechanisms such as those that block posttranscriptional gene silencing are involved in movement of PLRV within plants and its mechanical transmission between plants.

A review of host major-gene resistance to potato viruses X, Y, A and V in potato: genes, genetics and mapped locations.

In view of modern developments in the technologies available for breeding potatoes for resistance to virus diseases, it is timely to review the host major genes that confer resistance, in Solanum species, to potato viruses X, Y, A and V (the viruses for which the resistance genes have been most extensively studied). Over the course of 60 years, many such genes in Solanum species have been characterized: a comprehensive list is presented. Inheritance studies are reviewed, including linkage studies and molecular mapping, and the positions of resistance genes mapped so far are listed. It is apparent from recent research that disease resistance genes are often clustered in particular regions of the chromosomes; the significance of these resistance gene clusters is discussed. The information presented will be useful for potato breeding, and for genetic and mapping studies and gene cloning.

Breeding virus resistant potatoes (Solanum tuberosum): a review of traditional and molecular approaches.

Tetraploid cultivated potato (Solanum tuberosum) is the World's fourth most important crop and has been subjected to much breeding effort, including the incorporation of resistance to viruses. Several new approaches, ideas and technologies have emerged recently that could affect the future direction of virus resistance breeding. Thus, there are new opportunities to harness molecular techniques in the form of linked molecular markers to speed up and simplify selection of host resistance genes. The practical application of pathogen-derived transgenic resistance has arrived with the first release of GM potatoes engineered for virus resistance in the USA. Recently, a cloned host virus resistance gene from potato has been shown to be effective when inserted into a potato cultivar lacking the gene. These and other developments offer great opportunities for improving virus resistance, and it is timely to consider these advances and consider the future direction of resistance breeding in potato. We review the sources of available resistance, conventional breeding methods, marker-assisted selection, somaclonal variation, pathogen-derived and other transgenic resistance, and transformation with cloned host genes. The relative merits of the different methods are discussed, and the likely direction of future developments is considered.

D-dimer assay for deep vein thrombosis: its role with colour Doppler sonography.

AIM: To evaluate the role of a negative D-dimer assay in the initial management of patients with clinically suspected deep venous thrombosis (DVT), using colour Doppler ultrasound as the primary diagnostic technique. MATERIALS AND METHODS: A double-blind prospective trial was performed on 143 patients with clinically suspected DVT. All patients underwent a D-dimer assay prior to anticoagulant therapy. DVT was confirmed or excluded by diagnostic colour Doppler ultrasound within 24 h of presentation. RESULTS: In nearly one-third of the cases (31.8%), Doppler ultrasound was positive. The D-dimer assay demonstrated a sensitivity of 97.7% with only one false-negative, but the specificity was low at 48.9% with 45 false-positive results. The positive predictive value for D-dimer assay was 48.8%, whilst the important negative predictive value was 98%. CONCLUSION: If D-dimer was used to screen for DVT, and patients with negative results were not imaged, then the imaging workload could be reduced by 35%. In this study one small calf vein thrombus would have been missed by adopting this practice.Bradley, M. (2000). Clinical Radiology 55, 525-527.

NIMA-related kinase 2 (Nek2), a cell-cycle-regulated protein kinase localized to centrosomes, is complexed to protein phosphatase 1.

The cell cycle-regulated protein serine/threonine NIMA-related kinase 2 (Nek2), which shows a predominant localization at centrosomes, is identified as a protein which interacts with protein phosphatase 1 (PP1) using the yeast two-hybrid system. Complex formation between Nek2 and PP1 is supported by co-precipitation of the two proteins using transfected expression constructs of Nek2 and the endogenous Nek2/PP1 proteins. The sequence KVHF in the C-terminal region of Nek2, which conforms to the consensus PP1-binding motif, is shown to be essential for the interaction of Nek2 with PP1. Nek2 activity increases with autophosphorylation and addition of phosphatase inhibitors and decreases in the presence of PP1. PP1 is a substrate for Nek2 and phosphorylation of PP1gamma(1) on two C-terminal sites reduces its phosphatase activity. The presence of a ternary complex containing centrosomal Nek2-associated protein (C-Nap1), Nek2 and PP1 has also been demonstrated, and C-Nap1 is shown to be a substrate for both Nek2 and PP1 in vitro and in cell extracts. The implications of kinase-phosphatase complex formation involving Nek2 and PP1 are discussed in terms of the coordination of centrosome separation with cell cycle progression.

The determination of a reference range for new platelet parameters produced by the Bayer ADVIA120 full blood count analyser.

One hundred and twenty-two samples from normal volunteers were processed for full blood count analysis through the Bayer ADVIA120. The samples were simultaneously tested using the marker CD62P to establish the absence of platelet activation. Samples were tested at two time intervals, 20 min and 3.5 h, to simulate the testing practice for routine and urgent samples in our laboratory. From these data the mean and reference ranges, with 95% confidence limits, for platelet parameters were calculated for the ADVIA120. The time difference for some platelet parameters was significant. Hence two ranges have been reported. The age or smoking habit of the volunteer did not significantly affect platelet results but there was a statistically significant sex difference in some platelet parameters.

Formate protects stationary-phase Escherichia coli and Salmonella cells from killing by a cationic antimicrobial peptide.

For a sustained infection, enteric bacterial pathogens must evade, resist or tolerate a variety of antimicrobial host defence peptides and proteins. We report here that specific organic acids protect stationary-phase Escherichia coli and Salmonella cells from killing by a potent antimicrobial peptide derived from the human bactericidal/permeability-increasing protein (BPI). BPI-derived peptide P2 rapidly halted oxygen consumption by stationary-phase cells preincubated with glucose, pyruvate or malate and caused a 109-fold drop in cell viability within 90 min of addition. In marked contrast, O2 consumption and viability were not significantly affected in stationary-phase cells preincubated with formate or succinate. Experiments with fdhH, fdoG, fdnG, selC and sdhO mutants indicate that protection by formate and succinate requires their oxidation by the Fdh-N formate dehydrogenase and succinate dehydrogenase respectively. Protection was also dependent on the BipA GTPase but did not require the RpoS sigma factor. We conclude that the primary lesion caused by this cationic peptide is not gross permeabilization of the bacterial cytoplasmic membrane but may involve specific disruption of the respiratory chain. Because P2 shares sequence similarity with a range of other antimicrobial peptides, its cytotoxic mechanism has broader significance. Additionally, protective quantities of formate are secreted by E. coli and Salmonella during growth suggesting that such compounds are important determinants of bacterial survival in the host.

Potato mop-top virus RNA can move long distance in the absence of coat protein: evidence from resistant, transgenic plants.

Transgenic expression of a translatable version of the Potato mop-top virus (PMTV) coat protein (CP) gene (encoded by RNA 3) in Nicotiana benthamiana prevented production of symptoms and infective virus particles. RNAs 1 and 2 accumulated in inoculated and systemic leaves but, apart from small amounts of CP transgene RNA transcript, no genomic-length RNA 3 was found. Crude leaf extracts from inoculated plants were not infective. However, when RNA extracts from such transgenic plants were inoculated to nontransgenic N. benthamiana and N. clevelandii, RNA 1 and RNA 2 replicated in systemic leaves of both species in the absence of RNA 3 and virus particles, but symptoms did not develop. We suggest that the triple-gene block proteins of PMTV (encoded by RNA 2) represent a class of long-distance RNA movement factors.