Search results for: Abigail Gragg

Authors: Sam Bartlett, Thomas Bacquart, Arul Murugan, Abigail Morris

Abstract:

Fuel cell electric vehicles provide the potential to decarbonise road transport, create new economic opportunities, diversify national energy supply, and significantly reduce the environmental impacts of road transport. A potential issue, however, is that the catalyst used at the fuel cell cathode is susceptible to degradation by impurities, especially sulphur-containing compounds. A recent European Directive (2014/94/EU) stipulates that, from November 2017, all hydrogen provided to fuel cell vehicles in Europe must comply with the hydrogen purity specifications listed in ISO 14687-2; this includes reactive and toxic chemicals such as ammonia and total sulphur-containing compounds. This requirement poses great analytical challenges due to the instability of some of these compounds in calibration gas standards at relatively low amount fractions and the difficulty associated with undertaking measurements of groups of compounds rather than individual compounds. Without the available reference materials and analytical infrastructure, hydrogen refuelling stations will not be able to demonstrate compliance to the ISO 14687 specifications. The hydrogen purity laboratory at NPL provides world leading, accredited purity measurements to allow hydrogen refuelling stations to evidence compliance to ISO 14687. Utilising state-of-the-art methods that have been developed by NPL’s hydrogen purity laboratory, including a novel method for measuring total sulphur compounds at 4 nmol/mol and a hydrogen impurity enrichment device, we provide the capabilities necessary to achieve these goals. An overview of these capabilities will be given in this paper. As part of the EMPIR Hydrogen co-normative project ‘Metrology for sustainable hydrogen energy applications’, NPL are developing a validated analytical methodology for the measurement of speciated sulphur-containing compounds in hydrogen at low amount fractions pmol/mol to nmol/mol) to allow identification and measurement of individual sulphur-containing impurities in real samples of hydrogen (opposed to a ‘total sulphur’ measurement). This is achieved by producing a suite of stable gravimetrically-prepared primary reference gas standards containing low amount fractions of sulphur-containing compounds (hydrogen sulphide, carbonyl sulphide, carbon disulphide, 2-methyl-2-propanethiol and tetrahydrothiophene have been selected for use in this study) to be used in conjunction with novel dynamic dilution facilities to enable generation of pmol/mol to nmol/mol level gas mixtures (a dynamic method is required as compounds at these levels would be unstable in gas cylinder mixtures). Method development and optimisation are performed using gas chromatographic techniques assisted by cryo-trapping technologies and coupled with sulphur chemiluminescence detection to allow improved qualitative and quantitative analyses of sulphur-containing impurities in hydrogen. The paper will review the state-of-the art gas standard preparation techniques, including the use and testing of dynamic dilution technologies for reactive chemical components in hydrogen. Method development will also be presented highlighting the advances in the measurement of speciated sulphur compounds in hydrogen at low amount fractions.

Keywords: gas chromatography, hydrogen purity, ISO 14687, sulphur chemiluminescence detector

Procedia PDF Downloads 186

Authors: Isaac Martin, Abigail Lark, Sandra Morales, Eric W. Alton, Jane C. Davies

Abstract:

Objectives: The cystic fibrosis (CF) lung is a unique microbiological niche, wherein harmful bacteria persist for many years despite antibiotic therapy. Pseudomonas aeruginosa (Pa), the major culprit leading to lung decline and increased mortality, thrives in the lungs of patients with CF due to several factors that have been linked with poor antibiotic performance. Our group is investigating alternative therapies including bacteriophage cocktails with which we have previously demonstrated efficacy against planktonic organisms. In this study, we explored the effects of a 4-phage cocktail on Pa grown in two different conditions, intended to mirror the CF lung: a) alongside standard antibiotic treatment in pre-formed biofilms (structures formed by Pa-secreted exopolysaccharides which provide both physical and cell division barriers to antimicrobials and host defenses and b) in an acidic environment postulated to be present in the CF airway due both to the primary defect in bicarbonate secretion and secondary effects of inflammation. Methods: 16 Pa strains from CF patients at the Royal Brompton Hospital were selected based on sensitivity to a) ceftazidime/ tobramycin and b) the phage cocktail in a conventional plaque assay. To assess efficacy of phage in biofilms, 96 well plates with Pa (5x10⁷ CFU/ ml) were incubated in static conditions, allowing adherent bacterial colonies to form for 24 hr. Ceftazidime and tobramycin (both at 2 × MIC) were added, +/- bacteriophage (4x10⁸ PFU/mL) for a further 24 hr. Cell viability and biomass were estimated using fluorescent resazurin and crystal violet assays, respectively. To evaluate the effect of pH, strains were grown planktonically in shaking 96 well plates at pH 6.0, 6.6, 7.0 and 7.5 with tobramycin or phage, at varying concentrations. Cell viability was quantified by fluorescent resazurin assay. Results: For the biofilm assay, treatment groups were compared with untreated controls and expressed as percent reduction in cell viability and biomass. Addition of the 4-phage cocktail resulted in a 1.3-fold reduction in cell viability and 1.7-fold reduction in biomass (p < 0.001) when compared to standard antibiotic treatment alone. Notably, there was a 50 ± 15% reduction in cell viability and 60 ± 12% reduction in biomass (95% CI) for the 4 biofilms demonstrating the most resistance to antibiotic treatment. 83% of strains tested (n=6) showed decreased bacterial killing by tobramycin at acidic pHs (p < 0.01). However, 25% of strains (n=12) showed improved phage killing at acidic pHs (p < 0.05), with none showing the pattern of reduced efficacy at acidic pH demonstrated by tobramycin. Conclusion: The 4-phage anti-Pa cocktail tested against Pa performs well in pre-formed biofilms and in acidic environments; two conditions intended to mimic the CF lung. To our knowledge, these are the first data looking at the effects of subtle pH changes on phage-mediated bacterial killing in the context of Pa infection. These findings contribute to a growing body of evidence supporting the use of nebulised lytic bacteriophage as a treatment in the context of lung infection.

Keywords: biofilm, cystic fibrosis, pH, Pseudomonas aeruginosa, lytic bacteriophage

Procedia PDF Downloads 151

Authors: Judy M. Obliosca, Olivia Vest, Sandra Poulos, Kelsi Smith, Tammy Ferguson, Abigail Powers Lott, Alicia K. Smith, Yang Xu, Christopher K. Tison

Abstract:

Post-Traumatic Stress Disorder (PTSD) is a mental health problem that people may develop after experiencing traumatic events such as combat, natural disasters, and major emotional challenges. Tragically, the number of military personnel with PTSD correlates directly with the number of veterans who attempt suicide, with the highest rate in the Army. Research has shown epigenetic risks in those who are prone to several psychiatric dysfunctions, particularly PTSD. Once initiated in response to trauma, epigenetic alterations in particular, the DNA methylation in the form of 5-methylcytosine (5mC) alters chromatin structure and represses gene expression. Current methods to detect DNA methylation, such as bisulfite-based genomic sequencing techniques, are laborious and have massive analysis workflow while still having high error rates. A faster and simpler detection method of high sensitivity and precision would be useful in a clinical setting to confirm potential PTSD etiologies, prevent other psychiatric disorders, and improve military health. A nano-enhanced Surface Plasmon Resonance imaging (SPRi)-based assay that simultaneously detects site-specific 5mC base (termed as PTSD base) in methylated genes related to PTSD is being developed. The arrays on a sensing chip were first constructed for parallel detection of PTSD bases using synthetic and genomic DNA (gDNA) samples. For the gDNA sample extracted from the whole blood of a PTSD patient, the sample was first digested using specific restriction enzymes, and fragments were denatured to obtain single-stranded methylated target genes (ssDNA). The resulting mixture of ssDNA was then injected into the assay platform, where targets were captured by specific DNA aptamer probes previously immobilized on the surface of a sensing chip. The PTSD bases in targets were detected by anti-5-methylcytosine antibody (anti-5mC), and the resulting signals were then enhanced by the universal nanoenhancer. Preliminary results showed successful detection of a PTSD base in a gDNA sample. Brighter spot images and higher delta values (control-subtracted reflectivity signal) relative to those of the control were observed. We also implemented the in-house surface activation system for detection and developed SPRi disposable chips. Multiplexed PTSD base detection of target methylated genes in blood DNA from PTSD patients of severity conditions (asymptomatic and severe) was conducted. This diagnostic capability being developed is a platform technology, and upon successful implementation for PTSD, it could be reconfigured for the study of a wide variety of neurological disorders such as traumatic brain injury, Alzheimer’s disease, schizophrenia, and Huntington's disease and can be extended to the analyses of other sample matrices such as urine and saliva.

Keywords: epigenetic assay, DNA methylation, PTSD, whole blood, multiplexing

Procedia PDF Downloads 75