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The VPS4 component of the ESCRT machinery plays an essential role in HPV infectious entry and capsid disassembly
Lawrence Banks, Colin Crump, Anna Gozdzicka-Jozefiak, Martina Bergant, David Pim, Paola Massimi, Justyna Broniarczyk, 2017, original scientific article

Abstract: Human Papillomavirus (HPV) infection involves multiple steps, from cell attachment, through endocytic trafficking towards the trans-Golgi network, and, ultimately, the entry into the nucleus during mitosis. An essential viral protein in infectious entry is the minor capsid protein L2, which engages different components of the endocytic sorting machinery during this process. The ESCRT machinery is one such component that seems to play an important role in the early stages of infection. Here we have analysed the role of specific ESCRT components in HPV infection, and we find an essential role for VPS4. Loss of VPS4 blocks infection with multiple PV types, suggesting an evolutionarily conserved critical step in infectious entry. Intriguingly, both L1 and L2 can interact with VPS4, and appear to be in complex with VPS4 during the early stages of virus infection. By using cell lines stably expressing a dominant-negative mutant form of VPS4, we also show that loss of VPS4 ATPase activity results in a marked delay in capsid uncoating, resulting in a defect in the endocytic transport of incoming PsVs. These results demonstrate that the ESCRT machinery, and in particular VPS4, plays a critical role in the early stages of PV infection.
Found in: osebi
Keywords: HPV, ESCRT machinery, infection
Published: 08.05.2017; Views: 1899; Downloads: 143
.pdf Fulltext (1,10 MB)

Toxins in microalgae
Tanja Batkovič, Mojca Zotler, Meta Križaj, Jan David, Dani Bratuž, Anže Kuraj, 2017, final research report

Abstract: The aim of the project is to develop a detection system for the toxic algae Alexandrium minutum which can be than used as part of biosensor. First, we will isolate a single-domain antibody from a pre-immune library, then subclone its sequence in different vectors and produce it. Finally, we will design alternative ELISA methods and choose the most suitable to quantify the microalgae in water samples.
Found in: osebi
Keywords: Alexandrium minutum, mikroalge, biosenzorji ELISA
Published: 02.11.2018; Views: 1110; Downloads: 0
.pdf Fulltext (5,00 MB)

Phosphorylation of HPV-16 L2 Contributes To Efficient Virus Infectious Entry
Robert L. Garcea, Michael P. Myers, MARTINA bERGANT MARUŠIČ, David Pim, Paola Massimi, Justyna Broniarczyk, Lawrence Banks, 2019, original scientific article

Abstract: The Human Papillomavirus (HPV) capsid comprises two viral proteins, L1 and L2, with the L2 component being essential to ensure efficient endocytic transport of incoming viral genomes. Several studies have previously reported that L1 and L2 are post-translationally modified, but it is uncertain whether these modifications affect HPV infectious entry. Using a proteomic screen, we identified a highly conserved phospho-acceptor site on the HPV-16 and BPV-1 L2 proteins. The phospho-modification of L2, and its presence in HPV pseudovirions (PsVs), was confirmed using anti-phospho L2-specific antibodies. Mutation of the phospho-acceptor sites of both HPV-16 and BPV-1 L2 resulted in the production of infectious virus particles, with no differences in efficiency of packaging the reporter DNA. However, these mutated PsVs showed marked defects in infectious entry. Further analysis revealed a defect in uncoating, characterized by a delay in the exposure of a conformational epitope on L1 that indicates capsid uncoating. This uncoating defect was accompanied by a delay in the proteolysis of both L1 and L2 in mutated HPV-16 PsVs. Taken together, these studies indicate that phosphorylation of L2 during virus assembly plays an important role in optimal uncoating of virions during infection, suggesting that phosphorylation of the viral capsid proteins contributes to infectious entry.
Found in: osebi
Keywords: HPV, L2, infection, protein phosphorylation
Published: 05.06.2019; Views: 964; Downloads: 0
.pdf Fulltext (1,70 MB)

Atmospheric chemistry and physics in the atmosphere of a developed megacity (London): An overview of the REPARTEE experiment and its conclusions
Gavin J Phillips, Carole Helfter, Chiara F Di Marco, Eiko Nemitz, Fay Davies, Janet F Barlow, Tyrone Dunbar, Iain R White, Dudley E Shallcross, Stephen J Henshaw, K Fredrik Peterson, Brian Davison, Damien Martin, Ben Langford, C Nicholas Hewitt, Stephen M Ball, Justin M Langridge, A K Benton, Roderick L Jones, Paul I Williams, John Whitehead, Martin W Gallagher, Claire Martin, James R Dorsey, Hugh Coe, James D Allan, William J Bloss, Alistair J Thorpe, David C S Beddows, Manuel DallOsto, Roy M Harrison, Steven Smith, 2012, review article

Abstract: The Regents Park and Tower Environmental Experiment (REPARTEE) comprised two campaigns in London in October 2006 and October/November 2007. The experiment design involved measurements at a heavily trafficked roadside site, two urban background sites and an elevated site at 160-190 m above ground on the BT Tower, supplemented in the second campaign by Doppler lidar measurements of atmospheric vertical structure. A wide range of measurements of airborne particle physical metrics and chemical composition were made as well as measurements of a considerable range of gas phase species and the fluxes of both particulate and gas phase substances. Significant findings include (a) demonstration of the evaporation of traffic-generated nanoparticles during both horizontal and vertical atmospheric transport; (b) generation of a large base of information on the fluxes of nanoparticles, accumulation mode particles and specific chemical components of the aerosol and a range of gas phase species, as well as the elucidation of key processes and comparison with emissions inventories; (c) quantification of vertical gradients in selected aerosol and trace gas species which has demonstrated the important role of regional transport in influencing concentrations of sulphate, nitrate and secondary organic compounds within the atmosphere of London; (d) generation of new data on the atmospheric structure and turbulence above London, including the estimation of mixed layer depths; (e) provision of new data on trace gas dispersion in the urban atmosphere through the release of purposeful tracers; (f) the determination of spatial differences in aerosol particle size distributions and their interpretation in terms of sources and physico-chemical transformations; (g) studies of the nocturnal oxidation of nitrogen oxides and of the diurnal behaviour of nitrate aerosol in the urban atmosphere, and (h) new information on the chemical composition and source apportionment of particulate matter size fractions in the atmosphere of London derived both from bulk chemical analysis and aerosol mass spectrometry with two instrument types.
Found in: osebi
Keywords: megacity, trace gas, urban atmosphere, atmospheric transport, chemical composition, aerosol
Published: 18.07.2019; Views: 768; Downloads: 0
.pdf Fulltext (4,66 MB)

Breathomics and its Application for Disease Diagnosis: A Review of Analytical Techniques and Approaches
Enzo A Palombo, Konstantinos A Kouremenos, Ding Y Oh, Iain R White, David J Beale, Avinash V Karpe, Oliver A H Jones, 2018, independent scientific component part or a chapter in a monograph

Abstract: The application of metabolomics to an ever-greater variety of sample types is a key focus of systems biology research. Recently, there has been a strong focus on applying these approaches toward the rapid analysis of metabolites found in non-invasively acquired samples, such as exhaled breath (also known as ‘breathomics’). The sampling process involved in collecting exhaled breath is nonintrusive and comparatively low-cost. It uses a series of globally approved methods and provides researchers with easy access to the metabolites secreted by the human body. Owing to its accuracy and rapid nature, metabolomic analysis of breath is a rapidly growing field that has proven effective in detecting and diagnosing the early stages of numerous diseases and infections. This review discusses the various collection and analysis methods currently applied in breathomics research. Some of the salient research completed in this field to date is also assessed and discussed in order to provide a basis for possible future scientific directions.
Found in: osebi
Keywords: Metabolomics, breath research, VOCs, breathomics
Published: 22.07.2019; Views: 818; Downloads: 0
.pdf Fulltext (25,96 MB)

Circadian rhythm of exhaled biomarkers in health and asthma
Stephen Fowler, Royston Goodacre, David Ray, Dave Singh, Iain White, John Blaikley, Andrew Loudon, Robert Maidstone, Max Wilkinson, Hannah Durrington, 2019, original scientific article

Abstract: Circadian rhythms control many biological processes in the body in both health and disease. Greater understanding of diurnal variability in disease related biomarkers is crucial for their application in clinical practice and biomarkers of circadian rhythm are required to facilitate further research into disturbed chronicity. To determine if fractional exhaled nitric oxide and breath volatile biomarkers vary rhythmically during the day in healthy and asthmatic individuals. Ten individuals with moderate, atopic asthma (on regular inhaled corticosteroids) and 10 healthy volunteers (all non-smokers) completed an overnight visit where their exhaled breath volatiles and forced exhaled nitric oxide levels were collected every 6 h. Breath volatiles were analysed using gas chromatography mass spectrometry, after trapping these volatiles on sorbent materials for thermal desorption. Nine breath volatiles (including acetone and isoprene) exhibit diurnal variation across all individuals. Furthermore the circadian pattern of several VOCs is altered in individuals with asthma and fractional exhaled nitric oxide is rhythmic in asthma but not in healthy controls. Markers of circadian rhythm can be identified in breath and may offer insight into circadian profiling to help treat disease. Additionally this work suggests that time of day must be controlled when designing future biomarker discovery studies. Further work is required with larger cohorts to validate and extend these findings.
Found in: osebi
Keywords: VOCs, breath, asthma, circadian
Published: 21.10.2019; Views: 781; Downloads: 0
.pdf Fulltext (479,14 KB)

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