Validation of a method for the determination of the sensible-heat flux with Sodar data in free convection cases

A simple method to determine the value of the ground sensible-heat flux using Sodar data is presented and validated. The measurement of the variance of the wind velocity components gives us an estimate of the intensity of the atmospheric turbulence; the local value of variance of the vertical wind velocity s2w depends on the efficiency of thermal and mechanical turbulence production. The portion of the atmospheric boundary layer, where turbulent kinetic energy is prevalently produced by buoyancy forces, is characterised by profiles of s3w Oz and of (sensible-) heat flux which decrease linearly with height. The extrapolation to the ground of the former profile gives an estimate of the value of sensible-heat flux at the surface. The validation of the results is performed by comparison of the energy involved in the development of convective episodes calculated, over the same time interval, from sensible-heat flux at the surface with that derived from potential temperature profiles relative to two successive radio soundings. When perturbative processes like, for example, rise up of breezes, are absent, the estimates of energies are in excellent agreement, being the angular coefficient of regression line 1.01 and the linear correlation coefficient 0.93

A Whispering Gallery Mode based sensor platform for single enzyme real time conformational changes

Understanding the conformational changes that occur in an enzyme during turnover is crucial for understanding biomolecular processes and for new rational drug discovery. Many single-molecule techniques have been developed in the past three decades, such as Förster resonance energy transfer, although powerful, these techniques have their limitations, for example, limited temporal resolution, or necessity for fluorescent labelling. In this study a new class of highly sensitive optical devices based on the whispering gallery mode sensor have been investigated and employed to track the conformational changes of a kinase enzyme in real time. A thermostable Geobacillus stearothermophilus phosphoglycerate kinase enzyme has been used as model enzyme to prove that the optoplasmonic whispering gallery mode sensor can detect the conformational changes involved in enzyme turnover. The enzyme was expressed and purified to a high degree of purity that was suitable for crystallographic studies as well as biochemical investigation. The purified enzyme has been used for kinetics and biophysical enzymatic characterisation. A reproducible protocol has been established to immobilise the enzyme onto gold nanoparticles in a defined orientation to track the conformational changes occurring on the sensor. The enzyme has been immobilised onto the gold nanoparticles using two different methods: through the his-tags introduced onto the recombinant enzyme and directly onto the gold through a surface cysteine residue introduced by site-directed mutagenesis. The kinase enzyme was found to retain activity using both methods. To rationalise the observed conformational changes occurring during enzymatic turnover, the crystal structures of the native enzyme and its complexes with the substrate, and with substrate together with non-hydrolysable ATP, have been determined to high resolution, between 1.2 Å and 2 Å. Finally, for the first-time a real-time visualisation of the movement of the phosphoglycerate kinase during enzymatic turnover has been recorded. Repeating signals from the sensor were registered and only observed when both the substrate 3-phosphoglycerate and ATP are introduced with the enzyme into the sensor chamber. No signal is observed with the 3-phosphoglycerate alone or ATP alone or when 3-phosphoglycerate and a non-hydrolysable ATP analogue were used with the enzyme. The sensor system presented in this thesis shows potential for future fast, real-time, rapid throughput, lab-on-chip measurements for studying single enzymes.Engineering and Physical Sciences Research Council (EPSRC

Whispering-gallery mode (WGM) sensors: review of established and WGM-based techniques to study protein conformational dynamics

This is the final version. Available on open access from Elsevier via the DOI in this recordMonitoring the conformational dynamics of proteins is crucial for a better understanding of their biological functions. To observe the structural dynamics of proteins, it is often necessary to study each molecule individually. To this end, single-molecule techniques have been developed such as Förster resonance energy transfer and optical tweezers. However, although powerful, these techniques do have their limitations, for example, limited temporal resolution, or necessity for fluorescent labelling, and they can often only access a limited set of all protein motions. Here, within the context of established structural biology techniques, we review a new class of highly sensitive optical devices based on WGM, which characterise protein dynamics on previously inaccessible timescales, visualise motions throughout a protein, and track movements of single atoms.Engineering and Physical Sciences Research Council (EPSRC

Delphi consensus on the current clinical and therapeutic knowledge on Anderson-Fabry disease

BACKGROUND: Management of Anderson-Fabry disease (AFD) is contentious, particularly regarding enzyme replacement therapy (ERT). We report results of a Delphi consensus panel on AFD management. METHODS: A survey to gauge consensus among AFD experts was distributed online and responses were analysed. Statements on: 1) diagnosis; 2) when starting ERT; 3) management of ERT infusion and adverse reactions; and 4) follow-up/monitoring response to therapy and progression of disease were included. Responses without consensus were discussed with an enlarged panel and modified to reach consensus. RESULTS: 15 experts responded to the survey. After plenary discussion among the enlarged panel, consensus was reached on most statements. Key points were the use of a target organ biopsy to show Gb3 deposits in symptomatic women with negative molecular analysis, the need for ERT in symptomatic women and in all patients with persistent signs and symptoms±organ damage. It was agreed to assess vital signs before ERT administration and use a 0.2μL filter on infusion to reduce the risk of adverse reactions, that serum should be drawn prior to the first infusion for anti-agalsidase antibody analysis to have a baseline value if a subsequent adverse reaction appears, and that pre-medication is required in those with prior infusion reactions. Holter ECG monitoring, cardiac and brain MRI, renal parameters, and abdominal ultrasound were considered important for the assessment of disease progression and response at ERT. CONCLUSIONS: This consensus supplies guidance to healthcare providers on best practice in the management of patients with AFD and indicates a need for more guidanc

Expanding exploration of dynamic microplastic surface characteristics and interactions

This is the author accepted manuscript. The final version is available from Elsevier via the DOI in this recordMicroplastics have been found in all marine ecosystems, raising concern about their potential environmental impacts. Yet relatively little research has focused on surface characteristics, compared to polymer type. The aim of this review is to discuss the importance of microplastic surface properties and how expanded characterisation and more detailed quantification can aid in assessing their behaviours in aquatic environments. Concepts including surface roughness, formation of surface ecocoronae and sorptive behaviours of microplastic surfaces are discussed. To address these concepts, three exemplary methods are introduced and their application to the study of microplastic surfaces discussed with the following recommendations; atomic force microscopy should be explored for conducting physical surface characterisation and to examine surface roughness; double-shot Pyrolysis-Gas Chromatography–Mass Spectroscopy should be considered for examining microplastic sorption behaviours in multi-solute media; and finally, Whispering Gallery Mode nanosensing techniques should be explored as a potential means to generate data on microplastic sorption kinetics.University of ExeterUniversity of QueenslandNatural Environment Research Council (NERC

The Culture Environment Influences Both Gene Regulation and Phenotypic Heterogeneity in Escherichia coli

This is the final version of the article. Available from Frontiers Media via the DOI in this record.Microorganisms shape the composition of the medium they are growing in, which in turn has profound consequences on the reprogramming of the population gene-expression profile. In this paper, we investigate the progressive changes in pH and sugar availability in the medium of a growing Escherichia coli (E. coli) culture. We show how these changes have an effect on both the cellular heterogeneity within the microbial community and the gene-expression profile of the microbial population. We measure the changes in gene-expression as E. coli moves from lag, to exponential, and finally into stationary phase. We found that pathways linked to the changes in the medium composition such as ribosomal, tricarboxylic acid cycle (TCA), transport, and metabolism pathways are strongly regulated during the different growth phases. In order to quantify the corresponding temporal changes in the population heterogeneity, we measure the fraction of E. coli persisters surviving different antibiotic treatments during the various phases of growth. We show that the composition of the medium in which β-lactams or quinolones, but not aminoglycosides, are dissolved strongly affects the measured phenotypic heterogeneity within the culture. Our findings contribute to a better understanding on how the composition of the culture medium influences both the reprogramming in the population gene-expression and the emergence of phenotypic variants.This work was supported by a Royal Society Research Grant (RG140203), a Wellcome Trust Strategic Seed Corn Fund (WT097835/Z/11/Z), and a start up Grant from the University of Exeter awarded to SP. AS acknowledges support from the BBSRC through a SWBio-DTP studentship (BB/M009122/1). KP, KM, and PO would like to acknowledge support from the following awards: Wellcome Trust Institutional Strategic Support Fund (WT097835MF), Wellcome Trust Multi User Equipment Award (WT101650MA), and Medical Research Council Clinical Infrastructure Funding (MR/M008924/1). This work was partly supported by BBSRC award BB/1024631/1 to RT

Monophasic synovial sarcoma of the pharynx: a case report

Synovial sarcomas are a rare form of soft tissue sarcomas. We present a case of a 62 year-old male presenting with a left thyroid lump initially though to be a thyroid adenoma but subsequently diagnosed as a monophasic synovial sarcoma of the pharynx. We discuss the diagnosis and treatment of this case

Myocarditis following COVID-19 vaccine: incidence, presentation, diagnosis, pathophysiology, therapy, and outcomes put into perspective. A clinical consensus document supported by the Heart Failure Association of the European Society of Cardiology (ESC) and the ESC Working Group on Myocardial and Pericardial Diseases

Over 10 million doses of COVID-19 vaccines based on RNA technology, viral vectors, recombinant protein, and inactivated virus have been administered worldwide. Although generally very safe, post-vaccine myocarditis can result from adaptive humoral and cellular, cardiac-specific inflammation within days and weeks of vaccination. Rates of vaccine-associated myocarditis vary by age and sex with the highest rates in males between 12 and 39 years. The clinical course is generally mild with rare cases of left ventricular dysfunction, heart failure and arrhythmias. Mild cases are likely underdiagnosed as cardiac magnetic resonance imaging (CMR) is not commonly performed even in suspected cases and not at all in asymptomatic and mildly symptomatic patients. Hospitalization of symptomatic patients with electrocardiographic changes and increased plasma troponin levels is considered necessary in the acute phase to monitor for arrhythmias and potential decline in left ventricular function. In addition to evaluation for symptoms, electrocardiographic changes and elevated troponin levels, CMR is the best non-invasive diagnostic tool with endomyocardial biopsy being restricted to severe cases with heart failure and/or arrhythmias. The management beyond. guideline-directed treatment of heart failure and arrhythmias includes non-specific measures to control pain. Anti-inflammatory drugs such as non-steroidal anti-inflammatory drugs, and corticosteroids have been used in more severe cases, with only anecdotal evidence for their effectiveness. In all age groups studied, the overall risks of SARS-CoV-2 infection-related hospitalization and death are hugely greater than the risks from post-vaccine myocarditis. This consensus statement serves as a practical resource for physicians in their clinical practice, to understand, diagnose, and manage affected patients. Furthermore, it is intended to stimulate research in this area