Development of a novel UPLC-MS/MS-based platform to quantify amines, amino acids and methylarginines for applications in human disease phenotyping

Amine quantification is an important strategy in patient stratification and personalised medicine. This is because amines, including amino acids and methylarginines impact on many homeostatic processes. One important pathway regulated by amine levels is nitric oxide synthase (NOS). NOS is regulated by levels of (i) the substrate, arginine, (ii) amino acids which cycle with arginine and (iii) methylarginine inhibitors of NOS. However, biomarker research in this area is hindered by the lack of a unified analytical platform. Thus, the development of a common metabolomics platform, where a wide range of amino acids and methylarginines can be measured constitutes an important unmet need. Here we report a novel high-throughput ultra-high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) platform where ≈40 amine analytes, including arginine and methylarginines can be detected and quantified on a molar basis, in a single sample of human plasma. To validate the platform and to generate biomarkers, human plasma from a well-defined cohort of patients before and after coronary artery bypass surgery, who developed systemic inflammatory response syndrome (SIRS), were analysed. Bypass surgery with SIRS significantly altered 26 amine analytes, including arginine and ADMA. Consequently, pathway analysis revealed significant changes in a range of pathways including those associated with NOS

Metabolomic profiling of amines in sepsis predicts changes in NOS canonical pathways

Rationale Nitric oxide synthase (NOS) is a biomarker/target in sepsis. NOS activity is driven by amino acids, which cycle to regulate the substrate L-arginine in parallel with cycles which regulate the endogenous inhibitors ADMA and L-NMMA. The relationship between amines and the consequence of plasma changes on iNOS activity in early sepsis is not known. Objective Our objective was to apply a metabolomics approach to determine the influence of sepsis on a full array of amines and what consequence these changes may have on predicted iNOS activity. Methods and measurements 34 amino acids were measured using ultra purification mass spectrometry in the plasma of septic patients (n = 38) taken at the time of diagnosis and 24–72 hours post diagnosis and of healthy volunteers (n = 21). L-arginine and methylarginines were measured using liquid-chromatography mass spectrometry and ELISA. A top down approach was also taken to examine the most changed metabolic pathways by Ingenuity Pathway Analysis. The iNOS supporting capacity of plasma was determined using a mouse macrophage cell-based bioassay. Main results Of all the amines measured 22, including L-arginine and ADMA, displayed significant differences in samples from patients with sepsis. The functional consequence of increased ADMA and decreased L-arginine in context of all cumulative metabolic changes in plasma resulted in reduced iNOS supporting activity associated with sepsis. Conclusions In early sepsis profound changes in amine levels were defined by dominant changes in the iNOS canonical pathway resulting in functionally meaningful changes in the ability of plasma to regulate iNOS activity ex vivo

Pharmacological assessment of ibuprofen arginate on platelet aggregation and colon cancer cell killing

This work was funded in part by grants from the Wellcome Trust (0852551Z108/Z, to J.A.M.) and British Heart Foundation (FS/16/1/31699, to NSK). SM is a recipient of a PhD award from the King of Saud University, AT is a recipient of a MRC PhD studentship

Hydrodynamic slip can align thin nanoplatelets in shear flow

The large-scale processing of nanomaterials such as graphene and MoS2 relies on understanding the flow behaviour of nanometrically-thin platelets suspended in liquids. Here we show, by combining non-equilibrium molecular dynamics and continuum simulations, that rigid nanoplatelets can attain a stable orientation for sufficiently strong flows. Such a stable orientation is in contradiction with the rotational motion predicted by classical colloidal hydrodynamics. This surprising effect is due to hydrodynamic slip at the liquid-solid interface and occurs when the slip length is larger than the platelet thickness; a slip length of a few nanometers may be sufficient to observe alignment. The predictions we developed by examining pure and surface-modified graphene is applicable to different solvent/2D material combinations. The emergence of a fixed orientation in a direction nearly parallel to the flow implies a slip-dependent change in several macroscopic transport properties, with potential impact on applications ranging from functional inks to nanocomposites.Energy Technolog

A case study analysis of system efficiency, viability and energy values of SOFC based fuel cell Micro-CHP for office buildings

The built environment is a large contributor to the greenhouse gas emissions. The first step in tackling environmental impact and fuel cost reduction is the minimization of energy losses through introduction of new and more efficient energy conversion technologies. In this study, the potential application of fuel cell micro-CHP in a typical office building is investigated. A 1.5 kWel fuel cell micro-CHP has been installed in an office building at CIBSE headquarters (HQ). The unit converts natural gas to electricity, using Solid Oxide Fuel Cell (SOFC) stack. The installation and data from the operation of the system is discussed. The data shows that the heat recovered from the SOFC stack covers most of the domestic hot water requirement of the office building, requiring minimal top-up from the immersion heater.Publisher PD