'We don't need your blood': revealing the opportunities in the complex and surprisingly individualised volatile organic compound profiles of human skin and saliva

Non-invasive techniques for in-vivo sampling of volatile organic compounds (VOCs) from human skin and saliva have been developed and characterised using polydimethyl siloxane (PDMS) as a sorptive material. These samplers have been analysed by thermal desorption (TD) in combination with gas chromatography – mass spectrometry (GC-MS) and applied to global, untargeted metabolic profiling studies of skin and saliva. The utility of an existing PDMS ‘Skin Patch’ for untargeted metabolic profiling studies has been evaluated and background artefact levels and sample storage stability shown to be key considerations. Successful storage of skin samples for up to 21 days at -80 ºC has been demonstrated. Evaluation of published methods reveals that existing methods for saliva sampling do not address the physical–chemical attributes of VOCs and evidence is presented for adopting in situ sampling of salivary VOCs directly from the oral cavity. [Continues.

Extraordinarily high leaf selenium to sulfur ratios define ‘se-accumulator’ plants

Background and Aims: Selenium (Se) and sulfur (S) exhibit similar chemical properties. In flowering plants (angiosperms) selenate and sulfate are acquired and assimilated by common transport and metabolic pathways. It is hypothesized that most angiosperm species show little or no discrimination in the accumulation of Se and S in leaves when their roots are supplied a mixture of selenate and sulfate, but some, termed Se-accumulator plants, selectively accumulate Se in preference to S under these conditions. Methods: This paper surveys Se and S accumulation in leaves of 39 angiosperm species, chosen to represent the range of plant Se accumulation phenotypes, grown hydroponically under identical conditions. Results: The data show that, when supplied a mixture of selenate and sulfate: (1) plant species differ in both their leaf Se ([Se]leaf) and leaf S ([S]leaf) concentrations; (2) most angiosperms show little discrimination for the accumulation of Se and S in their leaves and, in non-accumulator plants, [Se]leaf and [S]leaf are highly correlated; (3) [Se]leaf in Se-accumulator plants is significantly greater than in other angiosperms, but [S]leaf, although high, is within the range expected for angiosperms in general; and (4) the Se/S quotient in leaves of Se-accumulator plants is significantly higher than in leaves of other angiosperms. Conclusion: The traits of extraordinarily high [Se]leaf and leaf Se/S quotients define the distinct elemental composition of Se-accumulator plants

Proteomic Analysis of a Noninvasive Human Model of Acute Inflammation and Its Resolution: The Twenty-one Day Gingivitis Model

The 21-day experimental gingivitis model, an established noninvasive model of inflammation in response to increasing bacterial accumulation in humans, is designed to enable the study of both the induction and resolution of inflammation. Here, we have analyzed gingival crevicular fluid, an oral fluid comprising a serum transudate and tissue exudates, by LC−MS/MS using Fourier transform ion cyclotron resonance mass spectrometry and iTRAQ isobaric mass tags, to establish meta-proteomic profiles of inflammation-induced changes in proteins in healthy young volunteers. Across the course of experimentally induced gingivitis, we identified 16 bacterial and 186 human proteins. Although abundances of the bacterial proteins identified did not vary temporally, Fusobacterium outer membrane proteins were detected. Fusobacterium species have previously been associated with periodontal health or disease. The human proteins identified spanned a wide range of compartments (both extracellular and intracellular) and functions, including serum proteins, proteins displaying antibacterial properties, and proteins with functions associated with cellular transcription, DNA binding, the cytoskeleton, cell adhesion, and cilia. PolySNAP3 clustering software was used in a multilayered analytical approach. Clusters of proteins that associated with changes to the clinical parameters included neuronal and synapse associated proteins

Use of a 3-item short-form version of the Barthel Index for use in stroke: systematic review and external validation

Background and Purpose—There may be a potential to reduce the number of items assessed in the Barthel Index (BI), and shortened versions of the BI have been described. We sought to collate all existing short-form BI (SF-BI) and perform a comparative validation using clinical trial data. Methods—We performed a systematic review across multidisciplinary electronic databases to find all published SF-BI. Our validation used the VISTA (Virtual International Stroke Trials Archive) resource. We describe concurrent validity (agreement of each SF-BI with BI), convergent and divergent validity (agreement of each SF-BI with other outcome measures available in the data set), predictive validity (association of prognostic factors with SF-BI outcomes), and content validity (item correlation and exploratory factor analyses). Results—From 3546 titles, we found 8 articles describing 6 differing SF-BI. Using acute trial data (n=8852), internal reliability suggested redundancy in BI (Cronbach α, 0.96). Each SF-BI demonstrated a strong correlation with BI, modified Rankin Scale, National Institutes of Health Stroke Scale (all ρ≥0.83; P<0.001). Using rehabilitation trial data (n=332), SF-BI demonstrated modest correlation with quality of life measures Stroke Impact Scale and 5 domain EuroQOL (ρ≥0.50, P<0.001). Prespecified prognostic factors were associated with SF-BI outcomes (all P<0.001). Our factor analysis described a 3 factor structure, and item reduction suggested an optimal 3-item SF-BI comprising bladder control, transfer, and mobility items in keeping with 1 of the 3-item SF-BI previously described in the literature. Conclusions—There is redundancy in the original BI; we have demonstrated internal and external validity of a 3-item SF-BI that should be simple to use

SLoMo: automated site localization of modifications from ETD/ECD mass spectra

Recently, software has become available to automate localization of phosphorylation sites from CID data and to assign associated confidence scores. We present an algorithm, SLoMo (Site Localization of Modifications), which extends this capability to ETD/ECD mass spectra. Furthermore, SLoMo caters for both high and low resolution data and allows for site-localization of any UniMod post-translational modification. SLoMo accepts input data from a variety of formats (e.g., Sequest, OMSSA). We validate SLoMo with high and low resolution ETD, ECD, and CID data

Sampling and characterisation of volatile organic compound profiles in human saliva using a polydimethylsiloxane coupon placed within the oral cavity

Evaluation of published methods reveals that existing methods for saliva sampling do not address the physical chemical attributes of volatile organic compounds (VOC). This study describes and presents evidence for adopting in-situ sampling of salivary VOC directly from the oral cavity using a polydimethylsiloxane (PDMS) based sampler. In-vitro studies indicated that the vapour pressure of analytes was a factor in both the recovery of analytes, and in the precision of the recovery. The highest recoveries were observed for VOC with the lowest vapour pressures, for example 5-nonanol (vapour pressure (Pv) = 14 PA) recoveries were approximately 20-times greater than those observed for octane (Pv = 1726 PA). Similarly, relative standard deviations reduced with vapour pressure, with the RSD for 5-nonanol responses observed to be 2.7 % to compared to RSD = 26 % for octane. Evaluation of VOC recovered from 6 in-vivo samples indicated that VOC concentrations in saliva may follow lognormal distributions; log-normal RSDs falling between 4.4% to 18.2% across the range of volatilities encountered. Increasing sampling time from 1 to 30 minutes indicated that the recovery of VOC into the sampler was effected by interaction between different physical chemical properties and biogenic flux. A sampling time of 10 min was found to offer an acceptable compromise that enabled a representative sample to be acquired for the widest range of observed VOC behaviours with the sampler. The potential to ‘tune’ the sampling protocol for targeted analysis based on these factors was also noted. Comparison with passive drool saliva collection revealed up to 105 enhancment with reduced variability compared to drooled samples. This approach to in-situ saliva sampling appears to have significant analytical utility for studying volatile signatures in humans

Changes in vitamin biomarkers during a 2-year intervention trial involving increased fruit and vegetable consumption by free-living volunteers

Peer reviewedPublisher PD

Using hormones to manage dairy cow fertility: the clinical and ethical beliefs of veterinary practitioners

In the face of a steady decline in dairy cow fertility over several decades, using hormones to assist reproduction has become common. In the European Union, hormones are prescription-only medicines, giving veterinary practitioners a central role in their deployment. This study explored the clinical and ethical beliefs of practitioners, and provides data on their current prescribing practices. During 2011, 93 practitioners working in England completed a questionnaire (95% response rate). Of the 714 non-organic farms they attended, only 4 farms (0.6%) never used hormones to assist the insemination of lactating dairy cows. Practitioners agreed (.80%) that hormones improve fertility and farm businesses profitability. They also agreed(.80%) that if farmers are able to tackle management issues contributing to poor oestrus expression, then over a five year period these outcomes would both improve, relative to using hormones instead. If management issues are addressed instead of prescribing hormones, practitioners envisaged a less favourable outcome for veterinary practices profitability (p,0.01), but an improvement in genetic selection for fertility (p,0.01) and overall cow welfare (p,0.01). On farms making no efforts to address underlying management problems, long-term routine use at the start of breeding for timing artificial insemination or inducing oestrus was judged ‘‘unacceptable’’ by 69% and 48% of practitioners, respectively. In contrast, practitioners agreed ($90%) that both these types of use are acceptable, provided a period of time has been allowed to elapse during which the cow is observed for natural oestrus. Issues discussed include: weighing quality versus length of cow life, fiscal factors, legal obligations, and balancing the interests of all stakeholders, including the increasing societal demand for food. This research fosters debate and critical appraisal, contributes to veterinary ethics, and encourages the pro-active development of professional codes of conduct

Native liquid extraction surface analysis mass spectrometry : analysis of noncovalent protein complexes directly from dried substrates

Liquid extraction surface analysis (LESA) mass spectrometry is a promising tool for the analysis of intact proteins from biological substrates. Here, we demonstrate native LESA mass spectrometry of noncovalent protein complexes of myoglobin and hemoglobin from a range of surfaces. Holomyoglobin, in which apomyoglobin is noncovalently bound to the prosthetic heme group, was observed following LESA mass spectrometry of myoglobin dried onto glass and polyvinylidene fluoride surfaces. Tetrameric hemoglobin [(αβ)(2)(4H)] was observed following LESA mass spectrometry of hemoglobin dried onto glass and polyvinylidene fluoride (PVDF) surfaces, and from dried blood spots (DBS) on filter paper. Heme-bound dimers and monomers were also observed. The ‘contact’ LESA approach was particularly suitable for the analysis of hemoglobin tetramers from DBS. [Figure: see text

Shoot yield drives phosphorus use efficiency in Brassica oleracea and correlates with root architecture traits

The environmental and financial costs of using inorganic phosphate fertilizers to maintain crop yield and quality are high. Breeding crops that acquire and use phosphorus (P) more efficiently could reduce these costs. The variation in shoot P concentration (shoot-P) and various measures of P use efficiency (PUE) were quantified among 355 Brassica oleracea L. accessions, 74 current commercial cultivars, and 90 doubled haploid (DH) mapping lines from a reference genetic mapping population. Accessions were grown at two or more external P concentrations in glasshouse experiments; commercial and DH accessions were also grown in replicated field experiments. Within the substantial species-wide diversity observed for shoot-P and various measures of PUE in B. oleracea, current commercial cultivars have greater PUE than would be expected by chance. This may be a consequence of breeding for increased yield, which is a significant component of most measures of PUE, or early establishment. Root development and architecture correlate with PUE; in particular, lateral root number, length, and growth rate. Significant quantitative trait loci associated with shoot-P and PUE occur on chromosomes C3 and C7. These data provide information to initiate breeding programmes to improve PUE in B. oleracea