Correlating instrumental and sensory analyses of flavour

The relationship between in vivo captured data from an atmospheric pressure chemical ionisation mass spectrometer (APCI-MS) and sensory/psychophysical analyses was investigated. The stimuli used were mainly single volatiles under gas phase control or calibration by development of different olfactometry methods. Gas phase concentration retronasal (via the mouth to the nasal cavity) and orthonasal (via the nostrils) thresholds were determined for a trained panel of 13 individuals. Four volatiles were used with different sensory/physico-chemical properties and an adapted staircase method was employed to measure the individual thresholds. The data showed good repeatability over short durations of one week and also longer ones of eight months. It was used to test the hypothesis that thresholds varied between people due to differences in their in-nose concentration as measured or estimated by the APCI-MS. The analysis did not support this theory but relationships between orthonasal and retronasal thresholds were shown, in which the latter were -50 times lower than the former. Threshold determination of a larger group of 20 individuals revealed clusters of individuals. Methods of producing square edged pulses of aroma compound in the gas phase were developed using a modified chromatograph autosampler with a gas flow of 5 mL. min 1 and pulse rate of 0.6 secs. A trained panel of 23 individuals performed two types of sensory test using pulsed and constant olfactometer outputs of isoamyl acetate. The original intention was to reveal whether pulsed odorants were perceived as the same as or different to constant concentration. Initial experiments yielded results that were difficult to interpret, although the nature of the results was clarified when simultaneous breath by breath analysis techniques were employed. Here it was shown that each individual in different repetitions disrupted the olfactometer output pattern in unpredictable ways. This pattern disruption was measured in two instrumental configurations, as either volatiles in an exhalation or volatiles as they were inhaled together with two types of sensory test. In both sensory tests the pattern of aromas in an inhalation revealed a relationship with perception. In particular, the sensory response in the time intensity study was related to differences in the inhalation profiles between people, which in turn was related to an individual's breathing. This shows that physiological differences such as breathing and the structure of the nasal cavity have an impact on perception

Correlating instrumental and sensory analyses of flavour

The relationship between in vivo captured data from an atmospheric pressure chemical ionisation mass spectrometer (APCI-MS) and sensory/psychophysical analyses was investigated. The stimuli used were mainly single volatiles under gas phase control or calibration by development of different olfactometry methods. Gas phase concentration retronasal (via the mouth to the nasal cavity) and orthonasal (via the nostrils) thresholds were determined for a trained panel of 13 individuals. Four volatiles were used with different sensory/physico-chemical properties and an adapted staircase method was employed to measure the individual thresholds. The data showed good repeatability over short durations of one week and also longer ones of eight months. It was used to test the hypothesis that thresholds varied between people due to differences in their in-nose concentration as measured or estimated by the APCI-MS. The analysis did not support this theory but relationships between orthonasal and retronasal thresholds were shown, in which the latter were -50 times lower than the former. Threshold determination of a larger group of 20 individuals revealed clusters of individuals. Methods of producing square edged pulses of aroma compound in the gas phase were developed using a modified chromatograph autosampler with a gas flow of 5 mL. min 1 and pulse rate of 0.6 secs. A trained panel of 23 individuals performed two types of sensory test using pulsed and constant olfactometer outputs of isoamyl acetate. The original intention was to reveal whether pulsed odorants were perceived as the same as or different to constant concentration. Initial experiments yielded results that were difficult to interpret, although the nature of the results was clarified when simultaneous breath by breath analysis techniques were employed. Here it was shown that each individual in different repetitions disrupted the olfactometer output pattern in unpredictable ways. This pattern disruption was measured in two instrumental configurations, as either volatiles in an exhalation or volatiles as they were inhaled together with two types of sensory test. In both sensory tests the pattern of aromas in an inhalation revealed a relationship with perception. In particular, the sensory response in the time intensity study was related to differences in the inhalation profiles between people, which in turn was related to an individual's breathing. This shows that physiological differences such as breathing and the structure of the nasal cavity have an impact on perception

Asymmetric Proteome Equalization of the Skeletal Muscle Proteome Using a Combinatorial Hexapeptide Library

Immobilized combinatorial peptide libraries have been advocated as a strategy for equalization of the dynamic range of a typical proteome. The technology has been applied predominantly to blood plasma and other biological fluids such as urine, but has not been used extensively to address the issue of dynamic range in tissue samples. Here, we have applied the combinatorial library approach to the equalization of a tissue where there is also a dramatic asymmetry in the range of abundances of proteins; namely, the soluble fraction of skeletal muscle. We have applied QconCAT and label-free methodology to the quantification of the proteins that bind to the beads as the loading is progressively increased. Although some equalization is achieved, and the most abundant proteins no longer dominate the proteome analysis, at high protein loadings a new asymmetry of protein expression is reached, consistent with the formation of complex assembles of heat shock proteins, cytoskeletal elements and other proteins on the beads. Loading at different ionic strength values leads to capture of different subpopulations of proteins, but does not completely eliminate the bias in protein accumulation. These assemblies may impair the broader utility of combinatorial library approaches to the equalization of tissue proteomes. However, the asymmetry in equalization is manifest at either low and high ionic strength values but manipulation of the solvent conditions may extend the capacity of the method

A comparison of collision cross section values obtained via travelling wave ion mobility-mass spectrometry and ultra high performance liquid chromatography-ion mobility-mass spectrometry : application to the characterisation of metabolites in rat urine

A comprehensive Collision Cross Section (CCS) library was obtained via Travelling Wave Ion Guide mobility measurements through direct infusion (DI). The library consists of CCS and Mass Spectral (MS) data in negative and positive ElectroSpray Ionisation (ESI) mode for 463 and 479 endogenous metabolites, respectively. For both ionisation modes combined, TWCCSN2 data were obtained for 542 non-redundant metabolites. These data were acquired on two different ion mobility enabled orthogonal acceleration QToF MS systems in two different laboratories, with the majority of the resulting TWCCSN2 values (from detected compounds) found to be within 1% of one another. Validation of these results against two independent, external TWCCSN2 data sources and predicted TWCCSN2 values indicated to be within 1-2% of these other values. The same metabolites were then analysed using a rapid reversed-phase ultra (high) performance liquid chromatographic (U(H)PLC) separation combined with IM and MS (IM-MS) thus providing retention time (tr), m/z and TWCCSN2 values (with the latter compared with the DI-IM-MS data). Analytes for which TWCCSN2 values were obtained by U(H)PLC-IM-MS showed good agreement with the results obtained from DI-IM-MS. The repeatability of the TWCCSN2 values obtained for these metabolites on the different ion mobility QToF systems, using either DI or LC, encouraged the further evaluation of the U(H)PLC-IM-MS approach via the analysis of samples of rat urine, from control and methotrexate-treated animals, in order to assess the potential of the approach for metabolite identification and profiling in metabolic phenotyping studies. Based on the database derived from the standards 63 metabolites were identified in rat urine, using positive ESI, based on the combination of tr, TWCCSN2 and MS data.</p

Specificity of the osmotic stress response in Candida albicans highlighted by quantitative proteomics

We are grateful to the BBSRC for funding the CRISP Consortium (Combinatorial Responses in Stress Pathways) under the SABR Initiative (Systems Approaches to Biological Research) (BB/F00513X/1; BB/F005210/1). AJPB was also funded by the BBSRC (BB/K017365/1), the ERC (C-2009-AdG-249793), the Wellcome Trust (097377), the MRC (MR/M026663/1), and the MRC Centre for Medical Mycology and the University of Aberdeen (MR/M026663/1).Peer reviewedPublisher PD

The chemical basis of serine palmitoyltransferase inhibition by myriocin

Sphingolipids (SLs) are essential components of cellular membranes formed from the condensation of L-serine and a long-chain acyl thioester. This first step is catalyzed by the pyridoxal-5'-phosphate (PLP)-dependent, enzyme serine palmitoyltransferase (SPT) which is a promising therapeutic target. The fungal natural product myriocin is a potent inhibitor of SPT and is widely used to block SL biosynthesis despite a lack of a detailed understanding of its molecular-mechanism. By combining spectroscopy, mass spectrometry, X-ray crystallography, and kinetics, we have characterized the molecular details of SPT inhibition by myriocin. Myriocin initially forms an external aldimine with PLP at the active site, and a structure of the resulting co-complex explains its nanomolar affinity for the enzyme. This co-complex then catalytically degrades via an unexpected 'retro-aldol-like' cleavage mechanism to a C18 aldehyde which in turn acts as a suicide inhibitor of SPT by covalent modification of the essential catalytic lysine. This surprising dual mechanism of inhibition rationalizes the extraordinary potency and longevity of myriocin inhibition.</p

Distinct Salmonella Enteritidis lineages associated with enterocolitis in high-income settings and invasive disease in low-income settings.

An epidemiological paradox surrounds Salmonella enterica serovar Enteritidis. In high-income settings, it has been responsible for an epidemic of poultry-associated, self-limiting enterocolitis, whereas in sub-Saharan Africa it is a major cause of invasive nontyphoidal Salmonella disease, associated with high case fatality. By whole-genome sequence analysis of 675 isolates of S. Enteritidis from 45 countries, we show the existence of a global epidemic clade and two new clades of S. Enteritidis that are geographically restricted to distinct regions of Africa. The African isolates display genomic degradation, a novel prophage repertoire, and an expanded multidrug resistance plasmid. S. Enteritidis is a further example of a Salmonella serotype that displays niche plasticity, with distinct clades that enable it to become a prominent cause of gastroenteritis in association with the industrial production of eggs and of multidrug-resistant, bloodstream-invasive infection in Africa.This work was supported by the Wellcome Trust. We would like to thank the members of the Pathogen Informatics Team and the core sequencing teams at the Wellcome Trust Sanger Institute (Cambridge, UK). We are grateful to D. Harris for work in managing the sequence data

Oxidative discolouration in whole-head and cut lettuce: biochemical and environmental influences on a complex phenotype and potential breeding strategies to improve shelf-life

Lettuce discolouration is a key post-harvest trait. The major enzyme controlling oxidative discolouration has long been considered to be polyphenol oxidase (PPO) however, levels of PPO and subsequent development of discolouration symptoms have not always correlated. The predominance of a latent state of the enzyme in plant tissues combined with substrate activation and contemporaneous suicide inactivation mechanisms are considered as potential explanations for this phenomenon. Leaf tissue physical properties have been associated with subsequent discolouration and these may be influenced by variation in nutrient availability, especially excess nitrogen and head maturity at harvest. Mild calcium and irrigation stress has also been associated with a reduction in subsequent discolouration, although excess irrigation has been linked to increased discolouration potentially through leaf physical properties. These environmental factors, including high temperature and UV light intensities, often have impacts on levels of phenolic compounds linking the environmental responses to the biochemistry of the PPO pathway. Breeding strategies targeting the PALand PPOpathway biochemistry and environmental response genes are discussed as a more cost-effective method of mitigating oxidative discolouration then either modified atmosphere packaging or post-harvest treatments, although current understanding of the biochemistry means that such programs are likely to be limited in nature and it is likely that they will need to be deployed alongside other methods for the foreseeable future

Meeting the challenges facing wheat production: The strategic research agenda of the Global Wheat Initiative

Wheat occupies a special role in global food security since, in addition to providing 20% of our carbohydrates and protein, almost 25% of the global production is traded internationally. The importance of wheat for food security was recognised by the Chief Agricultural Scientists of the G20 group of countries when they endorsed the establishment of the Wheat Initiative in 2011. The Wheat Initiative was tasked with supporting the wheat research community by facilitating collaboration, information and resource sharing and helping to build the capacity to address challenges facing production in an increasingly variable environment. Many countries invest in wheat research. Innovations in wheat breeding and agronomy have delivered enormous gains over the past few decades, with the average global yield increasing from just over 1 tonne per hectare in the early 1960s to around 3.5 tonnes in the past decade. These gains are threatened by climate change, the rapidly rising financial and environmental costs of fertilizer, and pesticides, combined with declines in water availability for irrigation in many regions. The international wheat research community has worked to identify major opportunities to help ensure that global wheat production can meet demand. The outcomes of these discussions are presented in this paper