Interactions between anionic radionuclides (129I, 79Se and 99Tc) and soil geocolloids

The aim of this work was to investigate the interactions of anionic radionuclides 129I, 77Se (as a proxy for 79Se) and 99Tc with soil geocolloids under a range of conditions. These anionic fission products are of specific concern to policy makers regarding human and environmental risk assessments. Previous research has demonstrated strong links between soil organic matter (SOM) content and reduced mobility of these radionuclides. Negatively charged humic substances (HS), such as humic acid (HA) and fulvic acid (FA), may constitute 80% of organic matter and the mechanisms that allow anionic radionuclide to interaction with these HSs are not well understood. In the case of all three radionuclides, speciation plays a significant role in controlling their environmental mobility, therefore HPLC and SEC coupled to ICP-MS was used to monitor the speciation changes as the isotopes were progressively incorporated into HA. X-ray absorption spectroscopy was also employed in order to establish the solid phase speciation of Se after reaction with soil geocolloids. Surface charge development of the HA significantly affected reaction with iodate (129IO3-) and iodide (129I-). Iodide added to HA systems demonstrated slow oxidation and formation of organically bound iodine (Org-129I) predominantly at higher pH (pH 6). Conversely IO3-, was rapidly transformed to form both I- and Org-I. As pH decreased, the rate of this reduction reaction increased. Increasing HA concentration also increased the rate of IO3- reduction and formation of Org-I. Previous research has suggested that the most likely mechanism is IO3- reduction to I2 or HOI which then binds with phenolic groups on OM forming Org-I species. However, IO3- was observed to rapidly bind to HA forming Org-I species with no initial evidence of I- formation; I- concentration then increased over time as Org-I decreased. Where Fe2+/Fe3+ was present increased reduction of IO3- to I- was observed, mediated by association with HA, resulting in less Org-I formation overall. Instantaneous reaction of I- with HA was observed in the presence of Fe2+/Fe3+, with bonding via cation bridging. Some I- was subsequently re-released as I- likely due to ongoing Fe hydrolysis. Modelling of the systems alone was successful and will assist the improvement of whole soil assemblage models. Selenite (Se(IV)) reaction with HA was most rapid at low pH, with minimal/no reduction occurring at > pH 6. Reduction of selenate (Se(VI)) also occurred but this was less than for Se(IV), at low pH. No formation of Se(VI) from Se(IV) was observed, suggesting no oxidation took place, however some formation of Se(IV) from Se(VI ) was observed, also the formation of an unknown Se species suspected to be organic in nature. Humic acid concentration had no significant effect on the rate of Se(IV) or Se(VI) reduction, suggesting that HA itself was not responsible for the reduction. X-ray absorption spectroscopy (XAS) demonstrated the potential for significant reduction to Se(0) at pH 4 and bonding through a Se-O-C chain. The role of microbial communities on Se(IV) and Se(VI) reduction in the HA systems was demonstrated through the use of soil inoculum and glucose additions in sterile and non-sterile systems. No reduction of Se(IV) or Se(VI) and bonding to HA was observed in filter and -irradiation systems. Additions of inoculum and glucose increased the rate of reduction. Additions of Fe2+ did not increase reduction of Se(IV) or Se(VI) when compared to non-sterile HA systems, however XAS analysis demonstrated formation of HA-Fe cation bridges. No reaction of pertechnetate (99Tc(VII)) with HA was observed in these aerobic systems. An unknown Tc species was occasionally observed (<0.005 µ L-1) and it is possible that this is an organic-Tc species. Significant incorporation of Tc into the solid phase was observed in aerobic soils, with most Tc(VII) being retained in soils with high OM contents and low pH. The mechanisms considered here build upon the basic processes considered in current biosphere models for I and Se. Assemblage models must be used in order to reliably model the interactions of elements within soils due to the complexity of the systems. In order to understand the long-term radiological risks associated with geological repositories, the fine-scale mechanisms must be understood geochemically across a range of different soil types and conditions. The effect of I and Se speciation on bioavailability in soils determines both the potential transfer of radioactive isotopes to the food chain from GDF’s and from aerial sources of contamination. Alongside this, the work also has significant implications for advising on cost-effect fertiliser application methods for both I and Se, in order to tackle nutrient deficiencies worldwide

Interactions between anionic radionuclides (129I, 79Se and 99Tc) and soil geocolloids

The aim of this work was to investigate the interactions of anionic radionuclides 129I, 77Se (as a proxy for 79Se) and 99Tc with soil geocolloids under a range of conditions. These anionic fission products are of specific concern to policy makers regarding human and environmental risk assessments. Previous research has demonstrated strong links between soil organic matter (SOM) content and reduced mobility of these radionuclides. Negatively charged humic substances (HS), such as humic acid (HA) and fulvic acid (FA), may constitute 80% of organic matter and the mechanisms that allow anionic radionuclide to interaction with these HSs are not well understood. In the case of all three radionuclides, speciation plays a significant role in controlling their environmental mobility, therefore HPLC and SEC coupled to ICP-MS was used to monitor the speciation changes as the isotopes were progressively incorporated into HA. X-ray absorption spectroscopy was also employed in order to establish the solid phase speciation of Se after reaction with soil geocolloids. Surface charge development of the HA significantly affected reaction with iodate (129IO3-) and iodide (129I-). Iodide added to HA systems demonstrated slow oxidation and formation of organically bound iodine (Org-129I) predominantly at higher pH (pH 6). Conversely IO3-, was rapidly transformed to form both I- and Org-I. As pH decreased, the rate of this reduction reaction increased. Increasing HA concentration also increased the rate of IO3- reduction and formation of Org-I. Previous research has suggested that the most likely mechanism is IO3- reduction to I2 or HOI which then binds with phenolic groups on OM forming Org-I species. However, IO3- was observed to rapidly bind to HA forming Org-I species with no initial evidence of I- formation; I- concentration then increased over time as Org-I decreased. Where Fe2+/Fe3+ was present increased reduction of IO3- to I- was observed, mediated by association with HA, resulting in less Org-I formation overall. Instantaneous reaction of I- with HA was observed in the presence of Fe2+/Fe3+, with bonding via cation bridging. Some I- was subsequently re-released as I- likely due to ongoing Fe hydrolysis. Modelling of the systems alone was successful and will assist the improvement of whole soil assemblage models. Selenite (Se(IV)) reaction with HA was most rapid at low pH, with minimal/no reduction occurring at > pH 6. Reduction of selenate (Se(VI)) also occurred but this was less than for Se(IV), at low pH. No formation of Se(VI) from Se(IV) was observed, suggesting no oxidation took place, however some formation of Se(IV) from Se(VI ) was observed, also the formation of an unknown Se species suspected to be organic in nature. Humic acid concentration had no significant effect on the rate of Se(IV) or Se(VI) reduction, suggesting that HA itself was not responsible for the reduction. X-ray absorption spectroscopy (XAS) demonstrated the potential for significant reduction to Se(0) at pH 4 and bonding through a Se-O-C chain. The role of microbial communities on Se(IV) and Se(VI) reduction in the HA systems was demonstrated through the use of soil inoculum and glucose additions in sterile and non-sterile systems. No reduction of Se(IV) or Se(VI) and bonding to HA was observed in filter and -irradiation systems. Additions of inoculum and glucose increased the rate of reduction. Additions of Fe2+ did not increase reduction of Se(IV) or Se(VI) when compared to non-sterile HA systems, however XAS analysis demonstrated formation of HA-Fe cation bridges. No reaction of pertechnetate (99Tc(VII)) with HA was observed in these aerobic systems. An unknown Tc species was occasionally observed (<0.005 µ L-1) and it is possible that this is an organic-Tc species. Significant incorporation of Tc into the solid phase was observed in aerobic soils, with most Tc(VII) being retained in soils with high OM contents and low pH. The mechanisms considered here build upon the basic processes considered in current biosphere models for I and Se. Assemblage models must be used in order to reliably model the interactions of elements within soils due to the complexity of the systems. In order to understand the long-term radiological risks associated with geological repositories, the fine-scale mechanisms must be understood geochemically across a range of different soil types and conditions. The effect of I and Se speciation on bioavailability in soils determines both the potential transfer of radioactive isotopes to the food chain from GDF’s and from aerial sources of contamination. Alongside this, the work also has significant implications for advising on cost-effect fertiliser application methods for both I and Se, in order to tackle nutrient deficiencies worldwide

Influences on the Design and Purification of Soluble, Recombinant Native-Like HIV-1 Envelope Glycoprotein Trimers

We have investigated factors that influence the production of native-like soluble, recombinant trimers based on the env genes of two isolates of human immunodeficiency virus type 1 (HIV-1), specifically 92UG037.8 (clade A) and CZA97.012 (clade C). When the recombinant trimers based on the env genes of isolates 92UG037.8 and CZA97.012 were made according to the SOSIP.664 design and purified by affinity chromatography using broadly neutralizing antibodies (bNAbs) against quaternary epitopes (PGT145 and PGT151, respectively), the resulting trimers are highly stable and they are fully native-like when visualized by negative-stain electron microscopy. They also have a native-like (i.e., abundant) oligomannose glycan composition and display multiple bNAb epitopes while occluding those for nonneutralizing antibodies. In contrast, uncleaved, histidine-tagged Foldon (Fd) domain-containing gp140 proteins (gp140UNC-Fd-His), based on the same env genes, very rarely form native-like trimers, a finding that is consistent with their antigenic and biophysical properties and glycan composition. The addition of a 20-residue flexible linker (FL20) between the gp120 and gp41 ectodomain (gp41ECTO) subunits to make the uncleaved 92UG037.8 gp140-FL20 construct is not sufficient to create a native-like trimer, but a small percentage of native-like trimers were produced when an I559P substitution in gp41ECTO was also present. The further addition of a disulfide bond (SOS) to link the gp120 and gp41 subunits in the uncleaved gp140-FL20-SOSIP protein increases native-like trimer formation to ∼20 to 30%. Analysis of the disulfide bond content shows that misfolded gp120 subunits are abundant in uncleaved CZA97.012 gp140UNC-Fd-His proteins but very rare in native-like trimer populations. The design and stabilization method and the purification strategy are, therefore, all important influences on the quality of trimeric Env proteins and hence their suitability as vaccine components

Kinetics of 99Tc speciation in aerobic soils

© 2019 The Authors Technetium-99 is a significant and long-lived component of spent nuclear fuel relevant to long-term assessments of radioactive waste disposal. Whilst 99Tc behaviour in poorly aerated environments is well known, the long-term bioavailability in aerobic soils following direct deposition or transport to the surface is less well understood. This work addresses two questions: (i) to what extent do soil properties control 99Tc kinetics in aerobic soils and (ii) over what experimental timescales must 99Tc kinetics be measured to make reliable long-term predictions of impact in the terrestrial environment? Soil microcosms spiked with 99TcO4− were incubated for 2.5 years and 99Tc transformations were periodically monitored by a sequential extraction, which enabled quantification of the reaction kinetics. Reduction in soluble 99Tc was slow and followed a double exponential kinetic model including a fast component enhanced by low pH, a slow component controlled by pH and organic matter, and a persistently soluble 99Tc fraction. Complexation with soil humus was key to the progressive immobilisation of 99Tc. Evidence for slow transfer to an unidentified ‘sink’ was found, with estimated decadal timeframes. Our data suggest that short-term experiments may not reliably predict long-term 99Tc solubility in soils with low to moderate organic matter contents

Disruption of the Opal Stop Codon Attenuates Chikungunya Virus-Induced Arthritis and Pathology

ABSTRACT Chikungunya virus (CHIKV) is a mosquito-borne alphavirus responsible for several significant outbreaks of debilitating acute and chronic arthritis and arthralgia over the past decade. These include a recent outbreak in the Caribbean islands and the Americas that caused more than 1 million cases of viral arthralgia. Despite the major impact of CHIKV on global health, viral determinants that promote CHIKV-induced disease are incompletely understood. Most CHIKV strains contain a conserved opal stop codon at the end of the viral nsP3 gene. However, CHIKV strains that encode an arginine codon in place of the opal stop codon have been described, and deep-sequencing analysis of a CHIKV isolate from the Caribbean identified both arginine and opal variants within this strain. Therefore, we hypothesized that the introduction of the arginine mutation in place of the opal termination codon may influence CHIKV virulence. We tested this by introducing the arginine mutation into a well-characterized infectious clone of a CHIKV strain from Sri Lanka and designated this virus Opal524R. This mutation did not impair viral replication kinetics in vitro or in vivo . Despite this, the Opal524R virus induced significantly less swelling, inflammation, and damage within the feet and ankles of infected mice. Further, we observed delayed induction of proinflammatory cytokines and chemokines, as well as reduced CD4 + T cell and NK cell recruitment compared to those in the parental strain. Therefore, the opal termination codon plays an important role in CHIKV pathogenesis, independently of effects on viral replication. IMPORTANCE Chikungunya virus (CHIKV) is a mosquito-borne alphavirus that causes significant outbreaks of viral arthralgia. Studies with CHIKV and other alphaviruses demonstrated that the opal termination codon within nsP3 is highly conserved. However, some strains of CHIKV and other alphaviruses contain mutations in the opal termination codon. These mutations alter the virulence of related alphaviruses in mammalian and mosquito hosts. Here, we report that a clinical isolate of a CHIKV strain from the recent outbreak in the Caribbean islands contains a mixture of viruses encoding either the opal termination codon or an arginine mutation. Mutating the opal stop codon to an arginine residue attenuates CHIKV-induced disease in a mouse model. Compared to infection with the opal-containing parental virus, infection with the arginine mutant causes limited swelling and inflammation, as well as dampened recruitment of immune mediators of pathology, including CD4 + T cells and NK cells. We propose that the opal termination codon plays an essential role in the induction of severe CHIKV disease

A global perspective on marine photosynthetic picoeukaryote community structure

A central goal in ecology is to understand the factors affecting the temporal dynamics and spatial distribution of microorganisms and the underlying processes causing differences in community structure and composition. However, little is known in this respect for photosynthetic picoeukaryotes (PPEs), algae that are now recognised as major players in marine CO2 fixation. Here, we analysed dot blot hybridisation and cloning–sequencing data, using the plastid-encoded 16S rRNA gene, from seven research cruises that encompassed all four ocean biomes. We provide insights into global abundance, α- and β-diversity distribution and the environmental factors shaping PPE community structure and composition. At the class level, the most commonly encountered PPEs were Prymnesiophyceae and Chrysophyceae. These taxa displayed complementary distribution patterns, with peak abundances of Prymnesiophyceae and Chrysophyceae in waters of high (25:1) or low (12:1) nitrogen:phosphorus (N:P) ratio, respectively. Significant differences in phylogenetic composition of PPEs were demonstrated for higher taxonomic levels between ocean basins, using Unifrac analyses of clone library sequence data. Differences in composition were generally greater between basins (interbasins) than within a basin (intrabasin). These differences were primarily linked to taxonomic variation in the composition of Prymnesiophyceae and Prasinophyceae whereas Chrysophyceae were phylogenetically similar in all libraries. These data provide better knowledge of PPE community structure across the world ocean and are crucial in assessing their evolution and contribution to CO2 fixation, especially in the context of global climate change

Improving the immunogenicity of native-like HIV-1 envelope trimers by hyperstabilization

The production of native-like recombinant versions of the HIV-1 envelope glycoprotein (Env) trimer requires overcoming the natural flexibility and instability of the complex. The engineered BG505 SOSIP.664 trimer mimics the structure and antigenicity of native Env. Here, we describe how the introduction of new disulfide bonds between the glycoprotein (gp)120 and gp41 subunits of SOSIP trimers of the BG505 and other genotypes improves their stability and antigenicity, reduces their conformational flexibility, and helps maintain them in the unliganded conformation. The resulting next-generation SOSIP.v5 trimers induce strong autologous tier-2 neutralizing antibody (NAb) responses in rabbits. In addition, the BG505 SOSIP.v6 trimers induced weak heterologous NAb responses against a subset of tier-2 viruses that were not elicited by the prototype BG505 SOSIP.664. These stabilization methods can be applied to trimers from multiple genotypes as components of multivalent vaccines aimed at inducing broadly NAbs (bNAbs)

Hundreds of variants clustered in genomic loci and biological pathways affect human height

Most common human traits and diseases have a polygenic pattern of inheritance: DNA sequence variants at many genetic loci influence the phenotype. Genome-wide association (GWA) studies have identified more than 600 variants associated with human traits, but these typically explain small fractions of phenotypic variation, raising questions about the use of further studies. Here, using 183,727 individuals, we show that hundreds of genetic variants, in at least 180 loci, influence adult height, a highly heritable and classic polygenic trait. The large number of loci reveals patterns with important implications for genetic studies of common human diseases and traits. First, the 180 loci are not random, but instead are enriched for genes that are connected in biological pathways (P = 0.016) and that underlie skeletal growth defects (P < 0.001). Second, the likely causal gene is often located near the most strongly associated variant: in 13 of 21 loci containing a known skeletal growth gene, that gene was closest to the associated variant. Third, at least 19 loci have multiple independently associated variants, suggesting that allelic heterogeneity is a frequent feature of polygenic traits, that comprehensive explorations of already-discovered loci should discover additional variants and that an appreciable fraction of associated loci may have been identified. Fourth, associated variants are enriched for likely functional effects on genes, being over-represented among variants that alter amino-acid structure of proteins and expression levels of nearby genes. Our data explain approximately 10% of the phenotypic variation in height, and we estimate that unidentified common variants of similar effect sizes would increase this figure to approximately 16% of phenotypic variation (approximately 20% of heritable variation). Although additional approaches are needed to dissect the genetic architecture of polygenic human traits fully, our findings indicate that GWA studies can identify large numbers of loci that implicate biologically relevant genes and pathways.

Development of a novel motivational interviewing (MI) informed peer-support intervention to support mothers to breastfeed for longer

Background: Many women in the UK stop breastfeeding before they would like to, and earlier than is recommended by the World Health Organization (WHO). Given the potential health benefits for mother and baby, new ways of supporting women to breastfeed for longer are required. The purpose of this study was to develop and characterise a novel Motivational Interviewing (MI) informed breastfeeding peer-support intervention. Methods: Qualitative interviews with health professionals and service providers (n=14), and focus groups with mothers (n=14), fathers (n=3), and breastfeeding peer-supporters (n=15) were carried out to understand experiences of breastfeeding peer-support and identify intervention options. Data were audio-recorded, transcribed, and analysed thematically. Consultation took place with a combined professional and lay Stakeholder Group (n=23). The Behaviour Change Wheel (BCW) guided intervention development process used the findings of the qualitative research and stakeholder consultation, alongside evidence from existing literature, to identify: the target behaviour to be changed; sources of this behaviour based on the Capability, Opportunity and Motivation (COM-B) model; intervention functions that could alter this behaviour; and; mode of delivery for the intervention. Behaviour change techniques included in the intervention were categorised using the Behaviour Change Technique Taxonomy Version 1 (BCTTv1). Results: Building knowledge, skills, confidence, and providing social support were perceived to be key functions of breastfeeding peer-support interventions that aim to decrease early discontinuation of breastfeeding. These features of breastfeeding peer-support mapped onto the BCW education, training, modelling and environmental restructuring intervention functions. Behaviour change techniques (BCTTv1) included social support, problem solving, and goal setting. The intervention included important inter-personal relational features (e.g. trust, honesty, kindness), and the BCTTv1 needed adaptation to incorporate this. Conclusions: The MI-informed breastfeeding peer-support intervention developed using this systematic and user-informed approach has a clear theoretical basis and well-described behaviour 3 change techniques. The process described could be useful in developing other complex interventions that incorporate peer-support and/or MI