Engineering Thermostability in Artificial Metalloenzymes to Increase Catalytic Activity

Protein engineering has shown widespread use in improving the industrial application of enzymes and broadening the conditions they are able to operate under by increasing their thermostability and solvent tolerance. Here, we show that protein engineering can be used to increase the thermostability of an artificial metalloenzyme. Thermostable variants of the human steroid carrier protein 2L, modified to bind a metal catalyst, were created by rational design using structural data and a 3DM database. These variants were tested to identify mutations that enhanced the stability of the protein scaffold, and a significant increase in melting temperature was observed with a number of modified metalloenzymes. The ability to withstand higher reaction temperatures resulted in an increased activity in the hydroformylation of 1-octene, with more than fivefold improvement in turnover number, whereas the selectivity for linear aldehyde remained high up to 80%

Sorptive removal of disinfection by-product precursors from UK lowland surface waters: impact of molecular weight and bromide

The current study compared the impact of three different unit processes, coagulation, granular activated carbon (GAC), and a novel suspended ion exchange (SIX) technology, on disinfection by-product formation potential (DBPFP) from two UK lowland water sources with medium to high bromide content. Specific attention was given to the influence of the organic molecular weight (MW) fraction on DBPFP as well as the impact of bromide concentration. Whilst few studies have investigated the impact of MW fractions from Liquid Chromatography with Organic Carbon Detection (LC-OCD) analysis on dissolved organic carbon (DOC) removal by different processes, none have studied the influence of DOC MW fractions from this analysis on DBP formation. The impact of higher bromide concentration was to decrease the total trihalomethane (THM) and haloacetic acid (HAA) mass concentration, in contrast to previously reported studies. Results indicated that for a moderate bromide concentration source (135 μg/L), the THM formation potential was reduced by 22% or 64% after coagulation or SIX treatment, respectively. For a high bromide content source (210 μg/L), the THM formation potential removal was 47% or 69% following GAC and SIX treatment, respectively. The trend was the same for HAAs, albeit with greater differences between the two processes/feedwaters with reference to overall removal. A statistical analysis indicated that organic matter of MW > 350 g/mol had a significant impact on DBPFP. A multiple linear regression of the MW fractions against DBPFP showed a strong correlation (R2 between 0.90 and 0.93), indicating that LC-OCD analysis alone could be used to predict DBP formation with reasonable accuracy, and offering the potential for rapid risk assessment of water source

Herschel-ATLAS: Blazars in the science demonstration phase field

To investigate the poorly constrained sub-mm counts and spectral properties of blazars we searched for these in the Herschel-ATLAS (H-ATLAS) science demonstration phase (SDP) survey catalog. We cross-matched 500 μm sources brighter than 50 mJy with the FIRST radio catalogue. We found two blazars, both previously known. Our study is among the first blind blazar searches at sub-mm wavelengths, i.e., in the spectral regime where little is still known about the blazar SEDs, but where the synchrotron peak of the most luminous blazars is expected to occur. Our early results are consistent with educated extrapolations of lower frequency counts and question indications of substantial spectral curvature downwardsand of spectral upturns at mm wavelengths. One of the two blazars is identified with a Fermi/LAT Υ-ray source and a WMAP source. The physical parameters of the two blazars are briefly discussed. These observations demonstrate that the H-ATLAS survey will provide key information aboutthe physics of blazars and their contribution to sub-mm counts.We are grateful to the referee, P. Padovani, for very useful comments. Thanks are due to the PACO collaboration for having made available the data on the 2 blazars. M.M. and L.B. thanks the staff at the Australia Telescope Compact Array site, Narrabri (NSW), for the valuable support they provide in running the telescope. Work partially supported by the Italian Space Agency (contract I/016/07/0 "COFIS'' and ASI/INAF Agreement I/072/09/0 for the Planck LFI Activity of Phase E2). This research has made use of the NASA/IPAC Extragalactic Database (NED) which is operated by the Jet Propulsion Laboratory, California Institute of Technology, under contract with the National Aeronautics and Space Administration

Modelling the impact of tidal range energy on species communities

Tidal energy has the potential to form a key component of the energy production in a number of countries, including the UK. Nonetheless, the deployment of tidal energy systems is associated with potential environmental impacts as prime resource sites often coincide with unique ecosystems inhabited by sensitive organisms. Previous studies have generally focused on the hydrodynamic impact of tidal energy schemes, i.e. how schemes alter the flow dynamics and sedimentary transport processes. Whilst these efforts are key in understanding environmental impacts, there is no straightforward step for translating sediment to faunal changes. Species distribution models offer methods to quantitatively predict certain possible impacts of tidal energy extraction. The River Severn is a distinguished candidate region for tidal energy in the UK featuring sites under stringent ecological protection regulations. We examine the impact of a proposed Severn tidal barrage on 14 species via the linking of hydrodynamic modelling to species distribution models. Through a selection of species that are linked via a simple food web system we extrapolate changes in prey species to the respective predator species. We show that species at lower trophic levels would be adversely affected by the barrage, but higher trophic level organisms increase in possible habitable area. Once food web relationships are acknowledged this increase in habitat area decreases, but is still net positive. Overall, all 14 species were affected, with most gaining in distribution area, and only four losing distribution area within the Severn Estuary. We conclude that a large-scale tidal barrage may have detrimental and complex impacts on species distribution, altering food web dynamics and altering food availability in the Severn Estuary. The methodology outlined herein can be transferred to the assessment and optimisation of prospective projects globally to aide in the sustainable introduction of the technology

Time-sculptures of Terrifying Ambiguity: Staging Inner Space and Migrating Realities in Analogue's Living Film Set

This article examines Analogue’s Living Film Set, an interactive theatre piece which uses miniature film sets, multi-touch surface technology and live video feeds to reframe my semi-remembered memories from the mid-1980s as a collective participatory experience. Drawing on new wave novelist J. G. Ballard’s notion of childhood memory as ‘time-sculptures of terrifying ambiguity’ [Ballard, J. G. 1963. “Time, Memory and Inner Space.” J. G. Ballard website (originally published in The Woman Journalist Magazine). Accessed August 6, 2015. http://www.jgballard.ca/non_fiction/jgb_time_memory_innerspace.html], I will demonstrate how my childhood town of Shepperton has been overwritten in both Ballardian literary fiction and the incursion of cinematic artifice from the neighbouring activities of Shepperton Film Studios. I argue that the ambiguity of my recollections and the contamination of my lived history with ‘prosthetic memories’ [Landsberg, Alison. 2004. Prosthetic Memory: The Transformation of American Remembrance in the Age of Mass Culture. New York, NY: Columbia UP, 20–21.] has provided a creative space to re-enact the blended hyperreality of my early childhood through the work’s intermedial form. I will conclude by examining how the shifting reality status of the media used within the performance intersects with the notion of ‘time-sculptures’ and problematises what Carol Martin [(2013). Theatre of the Real. Basingstoke: Palgrave Macmillan.] has identified as ‘theatre of the real’

Genotypic Diversity Is Associated with Clinical Outcome and Phenotype in Cryptococcal Meningitis across Southern Africa.

Cryptococcal meningitis is a major cause of mortality throughout the developing world, yet little is known about the genetic markers underlying Cryptococcal virulence and patient outcome. We studied a cohort of 230 Cryptococcus neoformans (Cn) isolates from HIV-positive South African clinical trial patients with detailed clinical follow-up using multi-locus sequence typing and in vitro phenotypic virulence assays, correlating these data with clinical and fungal markers of disease in the patient. South African Cn displayed high levels of genetic diversity and locus variability compared to globally distributed types, and we identified 50 sequence types grouped within the main molecular types VNI, VNII and VNB, with 72% of isolates typed into one of seven 'high frequency' sequence types. Spatial analysis of patients' cryptococcal genotype was not shown to be clustered geographically, which might argue against recent local acquisition and in favour of reactivation of latent infection. Through comparison of MLST genotyping data with clinical parameters, we found a relationship between genetic lineage and clinical outcome, with patients infected with the VNB lineage having significantly worse survival (n=8, HR 3.35, CI 1.51-7.20, p=0.003), and this was maintained even after adjustment for known prognostic indicators and treatment regimen. Comparison of fungal genotype with in vitro phenotype (phagocytosis, laccase activity and CSF survival) performed on a subset of 89 isolates revealed evidence of lineage-associated virulence phenotype, with the VNII lineage displaying increased laccase activity (p=0.001) and ex vivo CSF survival (p=0.0001). These findings show that Cryptococcus neoformans is a phenotypically heterogeneous pathogen, and that lineage plays an important role in cryptococcal virulence during human infection. Furthermore, a detailed understanding of the genetic diversity in Southern Africa will support further investigation into how genetic diversity is structured across African environments, allowing assessment of the risks different ecotypes pose to infection

De Novo VPS4A Mutations Cause Multisystem Disease with Abnormal Neurodevelopment.

The endosomal sorting complexes required for transport (ESCRTs) are essential for multiple membrane modeling and membrane-independent cellular processes. Here we describe six unrelated individuals with de novo missense variants affecting the ATPase domain of VPS4A, a critical enzyme regulating ESCRT function. Probands had structural brain abnormalities, severe neurodevelopmental delay, cataracts, growth impairment, and anemia. In cultured cells, overexpression of VPS4A mutants caused enlarged endosomal vacuoles resembling those induced by expression of known dominant-negative ATPase-defective forms of VPS4A. Proband-derived fibroblasts had enlarged endosomal structures with abnormal accumulation of the ESCRT protein IST1 on the limiting membrane. VPS4A function was also required for normal endosomal morphology and IST1 localization in iPSC-derived human neurons. Mutations affected other ESCRT-dependent cellular processes, including regulation of centrosome number, primary cilium morphology, nuclear membrane morphology, chromosome segregation, mitotic spindle formation, and cell cycle progression. We thus characterize a distinct multisystem disorder caused by mutations affecting VPS4A and demonstrate that its normal function is required for multiple human developmental and cellular processes.This work was supported by: UK Medical Research Council Project Grants [MR/M00046X/1], [MR/R026440/1] and Project grant from National Institute of Health Research Biomedical Research Centre at Addenbrooke's Hospital (to E.R.), Fondazione Bambino Gesù (Vite Coraggiose) and Italian Ministry of Health (CCR-2017-23669081) (to M.T.), National Institute for Health Research (NIHR) for the Cambridge Biomedical Research Centre and NIHR BioResource (Grant Number RG65966) (to F.L.R.), and a Sir Henry Dale Fellowship jointly funded by the Wellcome Trust and the Royal Society (Grant Number 216370/Z/19/Z) (to J.E.). CIMR was supported by a Wellcome Trust Strategic Award [100140] and Equipment Grant [093026]. This research was made possible through access to the data and findings generated by the 100,000 Genomes Project. The 100,000 Genomes Project is managed by Genomics England Limited (a wholly owned company of the Department of Health and Social Care). The 100,000 Genomes Project is funded by the National Institute for Health Research and NHS England. The Wellcome Trust, Cancer Research UK and the Medical Research Council have also funded research infrastructure. The 100,000 Genomes Project uses data provided by patients and collected by the National Health Service as part of their care and support

Cytomegalovirus distribution and evolution in hominines

Herpesviruses are thought to have evolved in very close association with their hosts. This is notably the case for cytomegaloviruses (CMVs; genus Cytomegalovirus) infecting primates, which exhibit a strong signal of co-divergence with their hosts. Some herpesviruses are however known to have crossed species barriers. Based on a limited sampling of CMV diversity in the hominine (African great ape and human) lineage, we hypothesized that chimpanzees and gorillas might have mutually exchanged CMVs in the past. Here, we performed a comprehensive molecular screening of all 9 African great ape species/subspecies, using 675 fecal samples collected from wild animals. We identified CMVs in eight species/subspecies, notably generating the first CMV sequences from bonobos. We used this extended dataset to test competing hypotheses with various degrees of co-divergence/number of host switches while simultaneously estimating the dates of these events in a Bayesian framework. The model best supported by the data involved the transmission of a gorilla CMV to the panine (chimpanzee and bonobo) lineage and the transmission of a panine CMV to the gorilla lineage prior to the divergence of chimpanzees and bonobos, more than 800,000 years ago. Panine CMVs then co-diverged with their hosts. These results add to a growing body of evidence suggesting that viruses with a double-stranded DNA genome (including other herpesviruses, adenoviruses, and papillomaviruses) often jumped between hominine lineages over the last few million years.Peer Reviewe

The Concise Guide to PHARMACOLOGY 2023/24:Catalytic receptors

The Concise Guide to PHARMACOLOGY 2023/24 is the sixth in this series of biennial publications. The Concise Guide provides concise overviews, mostly in tabular format, of the key properties of approximately 1800 drug targets, and nearly 6000 interactions with about 3900 ligands. There is an emphasis on selective pharmacology (where available), plus links to the open access knowledgebase source of drug targets and their ligands (https://www.guidetopharmacology.org/), which provides more detailed views of target and ligand properties. Although the Concise Guide constitutes almost 500 pages, the material presented is substantially reduced compared to information and links presented on the website. It provides a permanent, citable, point-in-time record that will survive database updates. The full contents of this section can be found at http://onlinelibrary.wiley.com/doi/10.1111/bph.16180. Catalytic receptors are one of the six major pharmacological targets into which the Guide is divided, with the others being: G protein-coupled receptors, ion channels, nuclear hormone receptors, enzymes and transporters. These are presented with nomenclature guidance and summary information on the best available pharmacological tools, alongside key references and suggestions for further reading. The landscape format of the Concise Guide is designed to facilitate comparison of related targets from material contemporary to mid-2023, and supersedes data presented in the 2021/22, 2019/20, 2017/18, 2015/16 and 2013/14 Concise Guides and previous Guides to Receptors and Channels. It is produced in close conjunction with the Nomenclature and Standards Committee of the International Union of Basic and Clinical Pharmacology (NC-IUPHAR), therefore, providing official IUPHAR classification and nomenclature for human drug targets, where appropriate.</p

Are differential consumption patterns in health-related behaviours an explanation for persistent and widening social inequalities in health in England?

During the last two decades, differential consumption patterns in health-related behaviours have increasingly been highlighted as playing an important role in explaining persistent and widening health inequalities. This period has also seen government public health policies in England place a greater emphasis on changing ‘lifestyle’ behaviours, in an attempt to tackle social inequalities in health. The aim of this study was to empirically examine the variation in health-related behaviour in relation to socio-economic position, in the English adult population, to determine the nature of this relationship and whether it has changed over time