Robotic multiwell planar patch-clamp for native and primary mammalian cells

Robotic multiwell planar patch-clamp has become common in drug development and safety programs because it enables efficient and systematic testing of compounds against ion channels during voltage-clamp. It has not, however, been adopted significantly in other important areas of ion channel research, where conventional patch-clamp remains the favored method. Here, we show the wider potential of the multiwell approach with the ability for efficient intracellular solution exchange, describing protocols and success rates for recording from a range of native and primary mammalian cells derived from blood vessels, arthritic joints and the immune and central nervous systems. The protocol involves preparing a suspension of single cells to be dispensed robotically into 4-8 microfluidic chambers each containing a glass chip with a small aperture. Under automated control, giga-seals and whole-cell access are achieved followed by preprogrammed routines of voltage paradigms and fast extracellular or intracellular solution exchange. Recording from 48 chambers usually takes 1-6 h depending on the experimental design and yields 16-33 cell recordings.

Response of the EAA FRSC to the EFRAG/ANC/FRC discussion paper: 'Towards a Disclosure Framework for the Notes'

We summarise the response of the EAA’s FRSC to Towards a Disclosure Framework for the Notes, a Discussion Paper (DP) issued jointly by EFRAG, ANC and FRC. While supportive of much of the DP, and in particular of the underlying aim to place disclosures on a sounder conceptual foundation, we identify two broad themes for further development. The first concerns the DP’s diagnosis of the problem, which is that the existing financial reporting is characterised by, on the one hand, disclosure overload and, on the other hand, an absence of a conceptual framework for organising and communicating disclosures. Our review of the literature suggests much greater support for the second of these two factors than for the first. The second broad theme is the purpose of the proposed DF, and the principles that are derived from this purpose. Here, we stress the need for the framework to better accommodate the context within which financial statement disclosures are used. In practice, this context is characterised by variation in information, incentives and enforcement, each of which has a considerable effect on the appropriate disclosure policy and practice in any given situation.

Does a structured gardening programme improve well-being In young-onset dementia? A preliminary study

Introduction: Young onset dementia (YOD) affects about 1 in 1500 people aged under 65 years in the UK. It is associated with loss of employment, independence and an increase in psychological distress. This project set out to identify the benefits of a 2 hour week) structured activity programme of gardening for people with YOD. Method: A mixed qualitative quantitative study of therapeutic gardening for people with YOD, measuring outcomes for both participants with YOD and their carers. 12 participants were recruited from a county wide older adults mental health service, based on onset of dementia being before 65 years of age(range 43-65 years). 2 dropped out and 1 died during the project. Measures included the Mini Mental State Examination, Bradford Well Being Profile, Large Allen Cognitive Level Screen and Pool Activity Level. Results: Over a one year period the carers of the people with YOD found that the project had given participants a renewed sense of purpose and increased well-being. while cognitive functioning declined. Conclusions: This study suggests that a meaningful guided activity programme can maintain or improve well-being in the presence of cognitive deterioration.

Bio-fortification of potato tubers using foliar zinc-fertiliser

Worldwide, many people are zinc (Zn)-deficient. Dietary Zn intake can be increased by producing crops with higher concentrations of Zn in their edible portions. This can be achieved by applying Zn-fertilisers to varieties with an increased ability to acquire Zn and to accumulate Zn in their edible portions. Potato (Solanum tuberosum L.) is an important food crop and is, therefore, a target for bio-fortification with Zn. Field trials incorporating a core collection of 23 potato genotypes, performed over 4 years (2006 – 2009), indicated significant genotypic effects on tuber Zn concentration and suggested that tuber Zn concentration was influenced by environmental effects, but also found that genotype environment (G E) interactions were not significant. Tuber Zn concentrations averaged 10.8 mg kg–1 dry matter (DM), and the ratio between the lowest and the highest varietal tuber Zn-concentration averaged 1.76. Tuber Zn concentrations could be increased by foliar Zn-fertilisation. Tuber yields of ‘Maris Piper’ were unaffected by foliar applications of < 1.08 g Zn plant–1. The relationship between tuber Zn concentration and foliar Zn application followed a saturation curve, reaching a maximum at approx. 30 mg Zn kg–1 DM at a foliar Zn application rate of 1.08 g plant–1. Despite a 40-fold increase in shoot Zn concentration compared to the unfertilised controls following foliar Zn fertilisation with 2.16 g Zn plant–1, only a doubling in tuber Zn concentration was observed. This suggests that the biofortification of tubers with Zn was restricted by the limited mobility of Zn in the phloem. A significant positive linear relationship between tuber Zn concentration and tuber N concentration supported the hypothesis of co-transport of Zn and N-compounds in the phloem.

Six new loci associated with body mass index highlight a neuronal influence on body weight regulation

Common variants at only two loci, FTO and MC4R, have been reproducibly associated with body mass index (BMI) in humans. To identify additional loci, we conducted meta-analysis of 15 genome-wide association studies for BMI (n > 32,000) and followed up top signals in 14 additional cohorts (n > 59,000). We strongly confirm FTO and MC4R and identify six additional loci (P < 5 x 10(-8)): TMEM18, KCTD15, GNPDA2, SH2B1, MTCH2 and NEGR1 (where a 45-kb deletion polymorphism is a candidate causal variant). Several of the likely causal genes are highly expressed or known to act in the central nervous system (CNS), emphasizing, as in rare monogenic forms of obesity, the role of the CNS in predisposition to obesity.

Association of vitamin D status with arterial blood pressure and hypertension risk: a mendelian randomisation study

BACKGROUND: Low plasma 25-hydroxyvitamin D (25[OH]D) concentration is associated with high arterial blood pressure and hypertension risk, but whether this association is causal is unknown. We used a mendelian randomisation approach to test whether 25(OH)D concentration is causally associated with blood pressure and hypertension risk. METHODS: In this mendelian randomisation study, we generated an allele score (25[OH]D synthesis score) based on variants of genes that affect 25(OH)D synthesis or substrate availability (CYP2R1 and DHCR7), which we used as a proxy for 25(OH)D concentration. We meta-analysed data for up to 108 173 individuals from 35 studies in the D-CarDia collaboration to investigate associations between the allele score and blood pressure measurements. We complemented these analyses with previously published summary statistics from the International Consortium on Blood Pressure (ICBP), the Cohorts for Heart and Aging Research in Genomic Epidemiology (CHARGE) consortium, and the Global Blood Pressure Genetics (Global BPGen) consortium. FINDINGS: In phenotypic analyses (up to n=49 363), increased 25(OH)D concentration was associated with decreased systolic blood pressure (β per 10% increase, -0·12 mm Hg, 95% CI -0·20 to -0·04; p=0·003) and reduced odds of hypertension (odds ratio [OR] 0·98, 95% CI 0·97-0·99; p=0·0003), but not with decreased diastolic blood pressure (β per 10% increase, -0·02 mm Hg, -0·08 to 0·03; p=0·37). In meta-analyses in which we combined data from D-CarDia and the ICBP (n=146 581, after exclusion of overlapping studies), each 25(OH)D-increasing allele of the synthesis score was associated with a change of -0·10 mm Hg in systolic blood pressure (-0·21 to -0·0001; p=0·0498) and a change of -0·08 mm Hg in diastolic blood pressure (-0·15 to -0·02; p=0·01). When D-CarDia and consortia data for hypertension were meta-analysed together (n=142 255), the synthesis score was associated with a reduced odds of hypertension (OR per allele, 0·98, 0·96-0·99; p=0·001). In instrumental variable analysis, each 10% increase in genetically instrumented 25(OH)D concentration was associated with a change of -0·29 mm Hg in diastolic blood pressure (-0·52 to -0·07; p=0·01), a change of -0·37 mm Hg in systolic blood pressure (-0·73 to 0·003; p=0·052), and an 8·1% decreased odds of hypertension (OR 0·92, 0·87-0·97; p=0·002). INTERPRETATION: Increased plasma concentrations of 25(OH)D might reduce the risk of hypertension. This finding warrants further investigation in an independent, similarly powered study.

Getting the measure of derivational morphology in adult speech a corpus analysis using MorphoQuantics

This paper describes the methodology used to compile a corpus called MorphoQuantics that contains a comprehensive set of 17,943 complex word types extracted from the spoken component of the British National Corpus (BNC). The categorisation of these complex words was derived primarily from the classification of Prefixes, Suffixes and Combining Forms proposed by Stein (2007). The MorphoQuantics corpus has been made available on a website of the same name; it lists 554 word-initial and 281 word-final morphemes in English, their etymology and meaning, and records the type and token frequencies of all the associated complex words containing these morphemes from the spoken element of the BNC, together with their Part of Speech. The results show that, although the number of word-initial affixes is nearly double that of word-final affixes, the relative number of each observed in the BNC is very similar; however, word-final affixes are more productive in that, on average, the frequency with which they attach to different bases is three times that of word-initial affixes. Finally, this paper considers how linguists, psycholinguists and psychologists may use MorphoQuantics to support their empirical work in first and second language acquisition, and clinical and educational research.

Finding vikings in the Danelaw

Historical, artefactual and place-name evidence indicates that Scandinavian migrants moved to eastern England in the ninth century AD, settling in the Danelaw. However, only a handful of characteristically Scandinavian burials have been found in the region. One, widely held, explanation is that most of these Scandinavian settlers quickly adopted local Christian burial customs, thus leaving Scandinavians indistinguishable from the Anglo-Saxon population. We undertook osteological and isotopic analysis to investigate the presence of first-generation Scandinavian migrants. Burials from Masham were typical of the later Anglo-Saxon period and included men, women and children. The location and positioning of the four adult burials from Coppergate, however, are unusual for Anglo-Scandinavian York. None of the skeletons revealed interpersonal violence. Isotopic evidence did not suggest a marine component in the diet of either group, but revealed migration on a regional, and possibly an international, scale. Combined strontium and oxygen isotope analysis should be used to investigate further both regional and Scandinavian migration in the later Anglo-Saxon period.

How reliable is internet-based self-reported identity, socio-demographic and obesity measures in European adults?

In e-health intervention studies, there are concerns about the reliability of internet-based, self-reported (SR) data and about the potential for identity fraud. This study introduced and tested a novel procedure for assessing the validity of internet-based, SR identity and validated anthropometric and demographic data via measurements performed face-to-face in a validation study (VS). Participants (n = 140) from seven European countries, participating in the Food4Me intervention study which aimed to test the efficacy of personalised nutrition approaches delivered via the internet, were invited to take part in the VS. Participants visited a research centre in each country within 2 weeks of providing SR data via the internet. Participants received detailed instructions on how to perform each measurement. Individual’s identity was checked visually and by repeated collection and analysis of buccal cell DNA for 33 genetic variants. Validation of identity using genomic information showed perfect concordance between SR and VS. Similar results were found for demographic data (age and sex verification). We observed strong intra-class correlation coefficients between SR and VS for anthropometric data (height 0.990, weight 0.994 and BMI 0.983). However, internet-based SR weight was under-reported (Δ −0.70 kg [−3.6 to 2.1], p < 0.0001) and, therefore, BMI was lower for SR data (Δ −0.29 kg m−2 [−1.5 to 1.0], p < 0.0001). BMI classification was correct in 93 % of cases. We demonstrate the utility of genotype information for detection of possible identity fraud in e-health studies and confirm the reliability of internet-based, SR anthropometric and demographic data collected in the Food4Me study.

Predicting fatty acid profiles in blood based on food intake and the FADS1 rs174546 SNP

SCOPE: A high intake of n-3 PUFA provides health benefits via changes in the n-6/n-3 ratio in blood. In addition to such dietary PUFAs, variants in the fatty acid desaturase 1 (FADS1) gene are also associated with altered PUFA profiles. METHODS AND RESULTS: We used mathematical modelling to predict levels of PUFA in whole blood, based on MHT and bolasso selected food items, anthropometric and lifestyle factors, and the rs174546 genotypes in FADS1 from 1,607 participants (Food4Me Study). The models were developed using data from the first reported time point (training set) and their predictive power was evaluated using data from the last reported time point (test set). Amongst other food items, fish, pizza, chicken and cereals were identified as being associated with the PUFA profiles. Using these food items and the rs174546 genotypes as predictors, models explained 26% to 43% of the variability in PUFA concentrations in the training set and 22% to 33% in the test set. CONCLUSIONS: Selecting food items using MHT is a valuable contribution to determine predictors, as our models' predictive power is higher compared to analogue studies. As unique feature, we additionally confirmed our models' power based on a test set.

Gender, childhood and children’s literature: the CIRCL approach

This paper explores the unique approach to childhood and children’s literature of the research and teaching of the ‘Graduate Centre for International Research in Childhood: Literature, Culture, Media (CIRCL)’. CIRCL follows in its work the arguments of UK critical theorist Jacqueline Rose in her seminal 1984 book The case of Peter Pan or the impossibility of children’s fiction. Rose’s work has been widely and routinely referenced in Children’s Literature studies particularly, but CIRCL interprets her arguments as having quite different implications than those usually assumed. Rose is generally attributed with having pointed out that ‘childhood’ is not one, homogenous category, but that childhood is divided by gender, race, and ethnic, political and religious (and so on) identities. But for CIRCL this is only a first step in Rose’s arguments and certainly one not unique to her work anyway: the perception of various cultural and historical childhoods is widely held. Instead, my paper explores how Rose’s arguments are centrally about how ‘childhood’ itself cannot be maintained in the face of division, a division, moreover, which operates inevitably at every level, and which derives from Rose’s interpretation of psychoanalysis as formulated by Sigmund Freud, which Rose derives in turn from her readings of the interpretations of Freud by French analyst Jacques Lacan and French critical theorist Jacques Derrida. Finally, my paper argues how Rose’s position is about any ‘identity’, including gender and that this allies her work closely to that of the famous gender theorist Judith Butler, whose arguments are often (mis) understood in the same ways as those of Rose.

Fish oil intakes providing dietary attainable levels of EPA and DHA reduces blood pressure in adults with systolic hypertension in a retrospective analysis

Background: Although a large number of randomized controlled trials (RCTs) have examined the impact of the n-3 (ω-3) fatty acids EPA (20:5n-3) and DHA (22:6n-3) on blood pressure and vascular function, the majority have used doses of EPA+DHA of > 3 g per d,which are unlikely to be achieved by diet manipulation. Objective: The objective was to examine, using a retrospective analysis from a multi-center RCT, the impact of recommended, dietary achievable EPA+DHA intakes on systolic and diastolic blood pressure and microvascular function in UK adults. Design: Healthy men and women (n = 312) completed a double-blind, placebo-controlled RCT consuming control oil, or fish oil providing 0.7 g or 1.8 g EPA+DHA per d in random order each for 8 wk. Fasting blood pressure and microvascular function (using Laser Doppler Iontophoresis) were assessed and plasma collected for the quantification of markers of vascular function. Participants were retrospectively genotyped for the eNOS rs1799983 variant. Results: No impact of n-3 fatty acid treatment or any treatment * eNOS genotype interactions were evident in the group as a whole for any of the clinical or biochemical outcomes. Assessment of response according to hypertension status at baseline indicated a significant (P=0.046) fish oil-induced reduction (mean 5 mmHg) in systolic blood pressure specifically in those with isolated systolic hypertension (n=31). No dose response was observed. Conclusions: These findings indicate that, in those with isolated systolic hypertension, daily doses of EPA+DHA as low as 0.7 g bring about clinically meaningful blood pressure reductions which, at a population level, would be associated with lower cardiovascular disease risk. Confirmation of findings in an RCT where participants are prospectively recruited on the basis of blood pressure status is required to draw definite conclusions. The Journal of Nutrition NUTRITION/2015/220475 Version 4