Comparison of two carbonaceous supported Fe-rich adsorbents for arsenate removal: A functionalisation and mechanistic study with applicability to groundwater treatment.

The presence of arsenic in groundwater, and through this in drinking water, has been shown to present a serious risk to public health in many regions of the world. In this study, two iron-rich carbonous adsorbents were compared for the removal of arsenate (As(V)) from groundwater. Biochars (FeO-biochar and FeO-pyrochar) derived from biomass waste were functionalised in two different ways with iron chloride for comparation. Batch and dynamic parameters were optimised to achieve >99% As(V) removal efficiency. Experimental data were best described by the pseudo-second order kinetic model, while multi-stage diffusion appeared to limit mass transfer of As(V). Among the isotherm models evaluated, the Freundlich model best described the experimental results with high correlation coefficients (R2 ≥ 0.94) for both adsorbents. Monolayer adsorption capacities were found to be 4.34 mg/g and 8.66 mg/g for FeO-biochar and FeO-pyrochar, respectively. Batch studies followed by instrumental characterisation of the materials indicated the removal mechanisms involved to be electrostatic interactions (outer-sphere), OH- ligand exchange (inner-sphere complexation) and hydrogen bonding with functional groups. Higher pHpzc (9.1), SBET (167.2 m2/g), and iron/elemental content for the FeO-pyrochar (compared with the FeO-biochar) suggested that both surface chemistry and porosity/surface area were important in adsorption. Dynamic studies showed FeO-pyrochar can be used to remove As(V) from groundwater even at low 'environmental' concentrations relevant to legislative limits (<10 µg/L), whereby 7 g of FeO-pyrochar was able to treat 5.4 L groundwater.

Streamlined Methods for Processing and Cryopreservation of Cell Therapy Products Using Automated Systems.

Streamlined procedures for processing and cryopreservation of cell therapies using good laboratory practices are integral to biomanufacturing process development and clinical applications. The protocol herein begins with the preparation of human cell types cultured as adherent (i.e., mesenchymal stromal cells, MSCs) or suspension cells (i.e., peripheral blood mononuclear cells, PBMCs) to comprehensively demonstrate procedures that are applicable to commonly used primary cell cultures. Cell processing steps consist of preparing high yields of cells for cryopreservation using instruments routinely used in cell manufacturing, including the Finia® Fill and Finish System and a controlled-rate freezer. The final steps comprise the storage of cells at subzero temperatures in liquid nitrogen vapor phase followed by the analysis of cell phenotypes before and after processing and cryopreservation, along with cell quality metrics for validation. Additionally, the protocol includes important considerations for the implementation of quality control measures for equipment operation and cell handling, as well as Good Laboratory Practices for cell manufacturing, which are essential for the translational use of cell therapies. Key features • The protocol applies to small- or large-scale manufacturing of cell therapy products. • It includes streamlined procedures for processing and cryopreservation of cells cultured as adherent cells (MSCs) and suspension cells (PBMCs). • Provides temperature control and rapid partitioning of sample in cryopreservation solution to maintain high viability of a range of cell types throughout the procedures. • This protocol employs the Finia® Fill and Finish System and a controlled-rate freezer. Graphical overview.

Pilot-scale phosphate recovery from wastewater to create a fertiliser product: An integrated assessment of adsorbent performance and quality.

Eutrophication and the predicted limited future availability of rock phosphate has triggered the increased development of phosphorus (P) recovery technologies, however, for remote regions, recovery solutions are still limited. Here, we report on a novel pilot-scale technology (FILTRAFLOTM-P reactor) to recover phosphate (PO43-) from wastewater effluent through a filtration/adsorption process in a rural setting. This unit employs enhanced gravitational filtration through adsorption media (here, a novel KOH deacetylated crab carapace based chitosan-calcite material (CCM)) with continuous self-backwashing. Trials were designed to assess how the FILTRAFLOTM-P unit would operate under 'real' conditions (both at low and high PO43- levels), and to ascertain the effectiveness of the adsorbent to recover phosphate from final effluent. High removal was achieved at low phosphate concentrations, bringing the residual effluent PO43- level below 1 mg/L (EU limit for sensitive water bodies), while phosphate was efficiently harvested (at more than 50%) at higher PO43- levels. Surface microprecipitation and inner-sphere complexation were postulated as the main PO43- adsorption mechanisms through XRD, XPS and EDX elemental mapping. Further, a quality assessment of the P-enriched CCM (which could be used as a potential soil amendment) was undertaken to consider elemental composition, microbiological assessment and quantification of organic micropollutants. Quality analysis indicated ∼2.5% P2O5 present, trace levels (well below legislative limits) of heavy metals and extremely low levels of organic pollutants (e.g., PCBs, pharmaceuticals). No detectable levels of target bacterial pathogens were observed. Pot trials showed that ryegrass cultivated with the addition of the CCM adsorbent achieved higher plant dry matter and P concentration when compared to unfertilised controls, with a slow-release kinetic pattern. This study showed that CCM used with the FILTRAFLOTM-P pilot reactor has high potential to recover phosphate from effluents and encourage resource recovery via bio-based management of waste.

Enhanced phosphate removal and potential recovery from wastewater by thermo-chemically calcinated shell adsorbents.

Shell from the seafood processing industry is an under-utilised waste resource worldwide. Calcite, the major component of shell is commonly used in wastewater treatment for the removal of phosphorus (P). Here, mussel and oyster shell-based adsorbents (MSB and OSB) were used for removal of P as phosphate (PO43-) from aqueous solution and secondary wastewater, following preparation through chemical calcination at 700 °C. Batch adsorption experiments were carried out to identify the effects of various operating parameters (e.g., pH, dosage, contact time, initial concentration of P ions, co-existing ions), while a desorption study helped to understand the availability of the bonded P. The optimal contact time for PO43- removal was 120 min using both adsorbents with the dose at 200 mg. Characterisation of the adsorbent was performed using SEM-EDX, pHpzc, BET, FTIR and XRD. The XRD analysis showed that both calcite and lime were present on the surface of the shell particles. P was adsorbed effectively through inner-sphere complexation and surface microprecipitation mechanisms, while an enhanced maximum P adsorption capacity of 12.44 mg/g for MSB and 8.25 mg/g for OSB was reached. The Redlich-Peterson isotherm model fitted well with the equilibrium isotherm data (R2 ≥ 0.97) which also suggested a heterogenic surface. The desorption study (on the saturated adsorbent) found that ~97% of bonded P could be plant available in soil. These results suggest that a shell-based adsorbent can serve as a promising material for P removal from real wastewater effluent and subsequently could be used as a soil conditioner.

Spatiotemporal trends and annual fluxes of pharmaceuticals in a Scottish priority catchment.

Pharmaceuticals (a class of emerging contaminants) are continuously introduced into effluent-receiving surface waters due to their incomplete removal within wastewater treatment plants (WWTPs). This work investigated the presence and distribution of eight commonly used human pharmaceuticals in the River Dee (Scotland, UK), a Scottish Environment Protection Agency priority catchment that is a conservation site and important raw water source. Grab sampling and passive sampling (Polar Organic Chemical Integrative Sampler, POCIS) was performed over 12 months, targeting: paracetamol, ibuprofen, and diclofenac (analgesics/anti-inflammatories); clarithromycin and trimethoprim (antibiotics); carbamazepine and fluoxetine (psychoactive drugs); and 17α-ethynylestradiol (estrogen hormone). Sampling sites spanned from the river's rural source to the heavily urbanised estuary into the North Sea. Ibuprofen (ranging 0.8-697 ng/L), paracetamol (ranging 4-658 ng/L), trimethoprim (ranging 3-505 ng/L), diclofenac (ranging 2-324 ng/L) and carbamazepine (ranging 1-222 ng/L) were consistently detected at the highest concentrations through grab sampling, with concentrations generally increasing down river with increasing urbanisation. However, POCIS revealed trace contamination of most compounds throughout the river (commonly <0.5 ng/L), indicating pollution may be related to diffuse sources. Analysis of river flows revealed that low flow and warm seasons corresponded to statistically significantly higher concentrations of diclofenac and carbamazepine, two compounds of environmental and regulatory concern. Below the largest WWTP, annual average fluxes ranged 0.1 kg/yr (clarithromycin) to 143.8 kg/yr (paracetamol), with 226.2 kg/yr for total target compounds. It was estimated that this source contributed >70% of the total mass loads (dissolved phase) of the target compounds in the river. As the River Dee is an important raw water source and conservation site, additional catchment monitoring is warranted to safeguard water quality and assess environmental risk of emerging contaminants, particularly in relation to unusual weather patterns, climate change and population growth.

A SPE-HPLC-MS/MS method for the simultaneous determination of prioritised pharmaceuticals and EDCs with high environmental risk potential in freshwater.

This work describes the development, optimisation and validation of an analytical method for the rapid determination of 17 priority pharmaceutical compounds and endocrine disrupting chemicals (EDCs). Rather than studying compounds from the same therapeutic class, the analyses aimed to determine target compounds with the highest risk potential (with particular regard to Scotland), providing a tool for further monitoring in different water matrices. Prioritisation was based on a systematic environmental risk assessment approach, using consumption data; wastewater treatment removal efficiency; environmental occurrence; toxicological effects; and pre-existing regulatory indicators. This process highlighted 17 compounds across various therapeutic classes, which were then quantified, at environmentally relevant concentrations, by a single analytical methodology. Analytical determination was achieved using a single-step solid phase extraction (SPE) procedure followed by high-performance liquid chromatography with tandem mass spectrometry (HPLC-MS/MS). The fully optimised method performed well for the majority of target compounds, with recoveries >71% for 15 of 17 analytes. The limits of quantification for most target analytes (14 of 17) ranged from 0.07 ng/L to 1.88 ng/L in river waters. The utility of this method was then demonstrated using real water samples associated with a rural hospital/setting. Eight compounds were targeted and detected, with the highest levels found for the analgesic, paracetamol (at up to 105,910 ng/L in the hospital discharge). This method offers a robust tool to monitor high priority pharmaceutical and EDC levels in various aqueous sample matrices.

Assessing hospital impact on pharmaceutical levels in a rural 'source-to-sink' water system.

It is widely recognised that inadequate removal of pharmaceuticals in wastewater may lead to their presence in surface waters. Hospitals are key point-sources for pharmaceuticals entering municipal waterways, and rural hospitals are of concern as receiving wastewater treatment plants (WWTPs) may be smaller, less advanced and thus less efficient. While most research has focused on urban settings, here we present results from a rural ''source-to-sink'' study around a hospital. The aim was to determine the contribution of pharmaceuticals discharged to a municipal wastewater system, and, to assess pharmaceutical removal efficiency in the WWTP. Samples were collected daily for one month to assess water quality and pharmaceuticals in the broader water cycle: (i) raw water supply; (ii) treated hospital tap water; (iii) hospital wastewater discharge; (iv) combined WWTP influent; and (v) final WWTP effluent. Target compounds included analgesics/antiinflammatories, antibiotics, psychiatric drugs, and a synthetic estrogen hormone. Concentrations ranged from: 3 ng/L (carbamazepine) to 105,910 ng/L (paracetamol) in hospital discharge; 5 ng/L (ibuprofen) to 105,780 ng/L (paracetamol) in WWTP influent; and 60 ng/L (clarithromycin) to 36,201 ng/L (paracetamol) in WWTP effluent. WWTP removal ranged from 87% (paracetamol) to <0% (carbamazepine and clarithromycin), and significant correlations with water quality characteristics and WWTP flow data were observed for some compounds. Results suggested that the hospital is an important source of certain pharmaceuticals entering municipal wastewater, and associated water quality parameters are impacted. Pharmaceutical persistence in the WWTP effluent highlighted the direct pathway these compounds have into receiving surface water, where their impact remains uncharacterised. Rural regions may face future challenges mitigating environmental risk as WWTP infrastructure ages, populations grow and pharmaceutical use and diversity continue to increase.

Reply to comments on "Low-cost chitosan-calcite adsorbent development for potential phosphate removal and recovery from wastewater effluent" by Pap et al. [Water research 173 (2020) 115573].

This letter is in response to the comments of Dr Hu and Dr Zhang on "Low-cost chitosan-calcite adsorbent development for potential phosphate removal and recovery from wastewater effluent" (Pap et al., 2020). We thank Dr Hu and Dr Zhang for their interest and comments, and having reflected, we wish to provide some clarification.

Low-cost chitosan-calcite adsorbent development for potential phosphate removal and recovery from wastewater effluent.

Phosphorous (P) recovery from wastewater will become increasingly vital in the future as terrestrial rock phosphate deposits are expended. Effective management of P as a critical resource will require new techniques to recover P from wastewater, ideally in a form that can be used in agriculture as fertiliser. In this study, batch and fixed-bed column conditions were tested using a novel KOH deacetylated calcite-chitosan based adsorbent (CCM) for P removal from aqueous solutions and wastewater effluents. The unique characteristics of this adsorbent as a phosphate adsorbent were the result of rich surface functionality (amine and sulphur functional groups of the chitosan and proteins) and the CaCO3 content (providing donor ligands; and additionally beneficial if the material were used as fertiliser, buffering soil acidification caused by nitrogen application). The maximum P adsorption capacity was determined to be 21.36 mgP/g (at 22 °C) and the endodermic process reached equilibrium after 120 min. The experimental data was best described using a Langmuir isotherm and a pseudo-second order kinetic model. The diffusion kinetic analysis highlighted the importance of both film and intraparticle mass-transport. Material characterisation suggested that the adsorption process involved interactions between P and functional groups (mostly -NH3+) due to electrostatic interaction on the chitosan chain or involved ligand exchange with CO32-. Analysis of materials using X-Ray Powder Diffraction (XRPD) and Thermogravimetric Analysis (TGA) indicated a microprecipitation-type mechanism may occur through the formation of hydroxylapatite (Ca5(PO4)3(OH)). Desorption studies demonstrated that the P-laden CCM (derived from crab carapace) had the potential to be reused in soil amendment as a slow-release P fertiliser. The effects of different operating parameters were explored in a fixed-bed column, and the experimental data fitted well to the Clark model (R2 = 0.99). The CCM also showed excellent P adsorption potential from secondary and final wastewater effluent in dynamic conditions, even at low P concentrations. Finally, a scale-up approach with cost analysis was used to evaluate the price and parameters needed for a potential large-scale P recovery system using this adsorbent.

Synthesis optimisation and characterisation of chitosan-calcite adsorbent from fishery-food waste for phosphorus removal.

Here, Box-Behnken design (BBD) approaches were utilised to optimise synthesis methodology for the chitosan-calcite rich adsorbent (CCM) made from fishery-food waste material (crab carapace), using low-temperature activation and potassium hydroxide (KOH). The effect of activation temperature, activation time and impregnation ratio was studied. The final adsorbent material was evaluated for its phosphorus (P) removal efficiency from liquid phase. Results showed that impregnation ratio was the most significant individual factor as this acted to increase surface deacetylation of the chitin (to chitosan) and increased the number of amine groups (-NH2) in the chitosan chain. P removal efficiency approached 75.89% (at initial P concentration of 20 mg/L) under optimised experimental conditions, i.e. where the impregnation ratio for KOH:carapace (g/g) was 1:1, the activation temperature was 105 °C and the activation time was 150 min. Predicted responses were in good agreement with the experimental data. Additionally, the pristine and CCM material were further analysed using scanning electron microscopy with energy dispersive X-ray spectroscopy (SEM/EDX), Brunauer-Emmett-Teller technique (BET), X-ray diffraction (XRD), Fourier transform infrared spectroscopy (FTIR) and thermal gravimetric analysis (TGA). Characterisation showed enhancements in surface chemistry (introducing positively charged amine groups), textural properties and thermal stability of the CCM.

A review of the potential utilisation of plastic waste as adsorbent for removal of hazardous priority contaminants from aqueous environments.

There is growing global awareness of the presence and negative impacts of waste plastic in the marine environment. Risks to wildlife include ingestion and entanglement for macro-plastic (larger than 5 mm in length), alongside food chain transfer for micro-plastics (less than 5 mm in length). Plastics in the marine environment have also been shown to adsorb and accumulate contaminants from seawater, e.g., heavy metals and hydrophobic organic compounds. This means that plastics can additionally act as vectors for transport of contaminants, permitting ecotoxicological risks to be spatially extended. However, the ability of waste plastic to adsorb pollutants also offers potential opportunity, if they can be used for the decontamination of wastewater. Here, we provide an overview of marine plastic types and distribution, and then systematically assess their potential to be repurposed as novel adsorbents. Data published in recent years are interrogated to gain an overview of the interaction mechanisms between marine plastics and both organic and inorganic contaminants. In addition, factors that may be exploited to enhance their performance in removal of contaminants are also reviewed and prioritised, e.g., surface modification and activation. This paper highlights the novel potential of repurposing plastic waste for wastewater treatment applications and seeks to identify key knowledge gaps and future research priorities for scientists and engineers.

Bone marrow donor selection and characterization of MSCs is critical for pre-clinical and clinical cell dose production.

BACKGROUND: Cell based therapies, such as bone marrow derived mesenchymal stem cells (BM-MSCs; also known as mesenchymal stromal cells), are currently under investigation for a number of disease applications. The current challenge facing the field is maintaining the consistency and quality of cells especially for cell dose production for pre-clinical testing and clinical trials. Here we determine how BM-donor variability and thus the derived MSCs factor into selection of the optimal primary cell lineage for cell production and testing in a pre-clinical swine model of trauma induced acute respiratory distress syndrome. METHODS: We harvested bone marrow and generated three different primary BM-MSCs from Yorkshire swine. Cells from these three donors were characterized based on (a) phenotype (morphology, differentiation capacity and flow cytometry), (b) in vitro growth kinetics and metabolic activity, and (c) functional analysis based on inhibition of lung endothelial cell permeability. RESULTS: Cells from each swine donor exhibited varied morphology, growth rate, and doubling times. All expressed the same magnitude of standard MSC cell surface markers by flow cytometry and had similar differentiation potential. Metabolic activity and growth potential at each of the passages varied between the three primary cell cultures. More importantly, the functional potency of the MSCs on inhibition of endothelial permeability was also cell donor dependent. CONCLUSION: This study suggests that for production of MSCs for cell-based therapy, it is imperative to examine donor variability and characterize derived MSCs for marker expression, growth and differentiation characteristics and testing potency in application dependent assays prior to selection of the optimal cell lineage for large scale expansion and dose production.

Comparison of sustainable biosorbents and ion-exchange resins to remove Sr2+ from simulant nuclear wastewater: Batch, dynamic and mechanism studies.

Removal of Sr2+ from aqueous media presents particular challenges, especially in complex wastes such as nuclear industry liquors. Commercial sorbents while effective, can be highly expensive and subject to negative effects from competing ions. Here we evaluate two potential biosorbents (crab carapace and spent distillery grain) as potential alternatives and compare their performance to two commercial sorbents for Sr2+ removal at industrially relevant concentrations (low mg/L). Physical and structural characterization of the materials was undertaken, and batch and dynamic studies were performed on Sr2+ solutions and simulated nuclear wastewater. Sorption performance was quantified with respect to contact time, initial concentration and ion-competition. Removal efficiencies were 20-70% for the biosorbents compared to 55-95% for the commercial materials. Results indicated sorption was predominantly through monolayer coverage on homogenous sites and could be described using a pseudo-second-order kinetic model. Studies with the simulant liquor showed Sr2+ sorption was reduced by 10-40% due to ion-competition for sites. Characterization of biosorbents before and after Sr2+ sorption suggested that outer-sphere complexation and ion-exchange were the primary Sr2+ removal mechanisms. The efficiency of crab carapace for Sr2+ removal from aqueous media (with adsorption capacity 3.92 mg/g.) at industrially relevant concentrations, together with its mechanical stability, implementation and disposal cost, makes it a competitive option compared to other biosorbents and commercial materials reported in the literature.

Elevated plasma levels of TIMP-3 are associated with a higher risk of acute respiratory distress syndrome and death following severe isolated traumatic brain injury.

BACKGROUND: Complications after injury, such as acute respiratory distress syndrome (ARDS), are common after traumatic brain injury (TBI) and associated with poor clinical outcomes. The mechanisms driving non-neurologic organ dysfunction after TBI are not well understood. Tissue inhibitor of matrix metalloproteinase-3 (TIMP-3) is a regulator of matrix metalloproteinase activity, inflammation, and vascular permeability, and hence has plausibility as a biomarker for the systemic response to TBI. METHODS: In a retrospective study of 182 patients with severe isolated TBI, we measured TIMP-3 in plasma obtained on emergency department arrival. We used non-parametric tests and logistic regression analyses to test the association of TIMP-3 with the incidence of ARDS within 8 days of admission and in-hospital mortality. RESULTS: TIMP-3 was significantly higher among subjects who developed ARDS compared with those who did not (median 2810 pg/mL vs. 2260 pg/mL, p=0.008), and significantly higher among subjects who died than among those who survived to discharge (median 2960 pg/mL vs. 2080 pg/mL, p<0.001). In an unadjusted logistic regression model, for each SD increase in plasma TIMP-3, the odds of ARDS increased significantly, OR 1.5 (95% CI 1.1 to 2.1). This association was only attenuated in multivariate models, OR 1.4 (95% CI 1.0 to 2.0). In an unadjusted logistic regression model, for each SD increase in plasma TIMP-3, the odds of death increased significantly, OR 1.7 (95% CI 1.2 to 2.3). The magnitude of this association was greater in a multivariate model adjusted for markers of injury severity, OR 1.9 (95% CI 1.2 to 2.8). DISCUSSION: TIMP-3 may play an important role in the biology of the systemic response to brain injury in humans. Along with clinical and demographic data, early measurements of plasma biomarkers such as TIMP-3 may help identify patients at higher risk of ARDS and death after severe isolated TBI. LEVEL OF EVIDENCE: III.

Concept for a hyperspectral remote sensing algorithm for floating marine macro plastics.

There is growing global concern over the chemical, biological and ecological impact of plastics in the ocean. Remote sensing has the potential to provide long-term, global monitoring but for marine plastics it is still in its early stages. Some progress has been made in hyperspectral remote sensing of marine macroplastics in the visible (VIS) to short wave infrared (SWIR) spectrum. We present a reflectance model of sunlight interacting with a sea surface littered with macro plastics, based on geometrical optics and the spectral signatures of plastic and seawater. This is a first step towards the development of a remote sensing algorithm for marine plastic using light reflectance measurements in air. Our model takes the colour, transparency, reflectivity and shape of plastic litter into account. This concept model can aid the design of laboratory, field and Earth observation measurements in the VIS-SWIR spectrum and explain the results.

Mesenchymal stem cell-derived extracellular vesicles attenuate pulmonary vascular permeability and lung injury induced by hemorrhagic shock and trauma.

BACKGROUND: Mesenchymal stem cells (MSCs) have been shown to mitigate vascular permeability in hemorrhagic shock (HS) and trauma-induced brain and lung injury. Mechanistically, paracrine factors secreted from MSCs have been identified that can recapitulate many of the potent biologic effects of MSCs in animal models of disease. Interestingly, MSC-derived extracellular vesicles (EVs), contain many of these key soluble factors, and have therapeutic potential independent of the parent cells. In this study we sought to determine whether MSC-derived EVs (MSC EVs) could recapitulate the beneficial therapeutic effects of MSCs on lung vascular permeability induced by HS in mice. METHODS: Mesenchymal stem cell EVs were isolated from human bone marrow-derived MSCs by ultracentrifugation. A mouse model of fixed pressure HS was used to study the effects of shock, shock + MSCs and shock + MSC EVs on lung vascular endothelial permeability. Mice were administered MSCs, MSC EVs, or saline IV. Lung tissue was harvested and assayed for permeability, RhoA/Rac1 activation, and for differential phosphoprotein expression. In vitro, human lung microvascular cells junctional integrity was evaluated by immunocytochemistry and endothelial cell impedance assays. RESULTS: Hemorrhagic shock-induced lung vascular permeability was significantly decreased by both MSC and MSC EV infusion. Phosphoprotein profiling of lung tissue revealed differential activation of proteins and pathways related to cytoskeletal rearrangement and regulation of vascular permeability by MSCs and MSC EVs. Lung tissue from treatment groups demonstrated decreased activation of the cytoskeletal GTPase RhoA. In vitro, human lung microvascular cells, MSC CM but not MSC-EVs prevented thrombin-induced endothelial cell permeability as measured by electrical cell-substrate impedance sensing system and immunocytochemistry of VE-cadherin and actin. CONCLUSION: Mesenchymal stem cells and MSC EVs modulate cytoskeletal signaling and attenuate lung vascular permeability after HS. Mesenchymal stem cell EVs may potentially be used as a novel "stem cell free" therapeutic to treat HS-induced lung injury.

Geochemical landscapes as drivers of trace and toxic element profiles in wild red deer (Cervus elaphus).

Tissue concentrations of essential trace and toxic elements in red deer (Cervus elaphus) are associated with the plants, soil and water they ingest. As such, variation in tissue concentrations is associated with variation in local geochemistry and bioavailability of elements. Physiological factors such as liver fluke (Fasciola hepatica) infection, breeding status, and in-tissue element interactions may also affect tissue concentrations, though their effects in red deer are not well understood. The primary objective of this study was therefore to survey wild red deer liver element concentrations across a range of geographically distinct populations during the Scottish red deer stalking season; and, in so doing, establishes element reference ranges while also exploring geographic and temporal variation and physiological factors. Livers were sampled from carcasses intended for human consumption on nine hunting estates during two seasons (2012-13, 2013-14). Samples were digested and analysed by ICP-OES for essential trace elements (Co, Cu, Fe, Mn, Mo, Se, Zn) and for Cd. Data (n=787) were modelled against cull location, date, and F. hepatica diagnosis. Interactions between elements within liver, and differences in element profiles between estates, were explored by principal component analysis. Our results revealed marked geographic variation in Cd, Cu and Se, where up to four-fold differences in median element concentrations occurred between estates, and, in males, Mn, Mo and Zn declined as the breeding season approached. In both sexes, within-liver associations (Cd-Cu-Se and Mn-Mo-Zn) were found. In females, liver Zn was greater on average in individuals that were not infected with F. hepatica. This study is the first to quantify geographic variation in Scottish red deer liver element concentrations; the drivers of which remain to be explored (and may be management related), and, the consequence of which may affect sub-clinical health.

Cytokines Elevated in HIV Elite Controllers Reduce HIV Replication In Vitro and Modulate HIV Restriction Factor Expression.

A subset of HIV-infected individuals termed elite controllers (ECs) maintain CD4+ T cell counts and control viral replication in the absence of antiretroviral therapy (ART). Systemic cytokine responses may differentiate ECs from subjects with uncontrolled viral replication or from those who require ART to suppress viral replication. We measured 87 cytokines in four groups of women: 73 ECs, 42 with pharmacologically suppressed viremia (ART), 42 with uncontrolled viral replication (noncontrollers [NCs]), and 48 HIV-uninfected (NEG) subjects. Four cytokines were elevated in ECs but not NCs or ART subjects: CCL14, CCL21, CCL27, and XCL1. In addition, median stromal cell-derived factor-1 (SDF-1) levels were 43% higher in ECs than in NCs. The combination of the five cytokines suppressed R5 and X4 virus replication in resting CD4+ T cells, and individually SDF-1ß, CCL14, and CCL27 suppressed R5 virus replication, while SDF-1ß, CCL21, and CCL14 suppressed X4 virus replication. Functional studies revealed that the combination of the five cytokines upregulated CD69 and CCR5 and downregulated CXCR4 and CCR7 on CD4+ T cells. The CD69 and CXCR4 effects were driven by SDF-1, while CCL21 downregulated CCR7. The combination of the EC-associated cytokines induced expression of the anti-HIV host restriction factors IFITM1 and IFITM2 and suppressed expression of RNase L and SAMHD1. These results identify a set of cytokines that are elevated in ECs and define their effects on cellular activation, HIV coreceptor expression, and innate restriction factor expression. This cytokine pattern may be a signature characteristic of HIV-1 elite control, potentially important for HIV therapeutic and curative strategies.IMPORTANCE Approximately 1% of people infected with HIV control virus replication without taking antiviral medications. These subjects, termed elite controllers (ECs), are known to have stronger immune responses targeting HIV than the typical HIV-infected subject, but the exact mechanisms of how their immune responses control infection are not known. In this study, we identified five soluble immune signaling molecules (cytokines) in the blood that were higher in ECs than in subjects with typical chronic HIV infection. We demonstrated that these cytokines can activate CD4+ T cells, the target cells for HIV infection. Furthermore, these five EC-associated cytokines could change expression levels of intrinsic resistance factors, or molecules inside the target cell that fight HIV infection. This study is significant in that it identified cytokines elevated in subjects with a good immune response against HIV and defined potential mechanisms as to how these cytokines could induce resistance to the virus in target cells.

Prevalence of Liver Fluke (Fasciola hepatica) in Wild Red Deer (Cervus elaphus): Coproantigen ELISA Is a Practicable Alternative to Faecal Egg Counting for Surveillance in Remote Populations.

Red deer (Cervus elaphus) are hosts of liver fluke (Fasciola hepatica); yet, prevalence is rarely quantified in wild populations. Testing fresh samples from remote regions by faecal examination (FE) can be logistically challenging; hence, we appraise frozen storage and the use of a coproantigen ELISA (cELISA) for F. hepatica surveillance. We also present cELISA surveillance data for red deer from the Highlands of Scotland. Diagnoses in faecal samples (207 frozen, 146 fresh) were compared using a cELISA and by FE. For each storage method (frozen or fresh), agreement between the two diagnostics was estimated at individual and population levels, where population prevalence was stratified into cohorts (e.g., by sampling location). To approximate sensitivity and specificity, 65 post-slaughter whole liver examinations were used as a reference. At the individual level, FE and cELISA diagnoses agreed moderately (κfrozen = 0.46; κfresh = 0.51), a likely reflection of their underlying principles. At the population level, FE and cELISA cohort prevalence correlated strongly (Pearson's R = 0.89, p < 0.0001), reflecting good agreement on relative differences between cohort prevalence. In frozen samples, prevalence by cELISA exceeded FE overall (42.8% vs. 25.8%) and in 9/12 cohorts, alluding to differences in sensitivity; though, in fresh samples, no significant difference was found. In 959 deer tested by cELISA across the Scottish Highlands, infection prevalence ranged from 9.6% to 53% by sampling location. We highlight two key advantages of cELISA over FE: i) the ability to store samples long term (frozen) without apparent loss in diagnostic power; and ii) reduced labour and the ability to process large batches. Further evaluation of cELISA sensitivity in red deer, where a range of fluke burdens can be obtained, is desirable. In the interim, the cELISA is a practicable diagnostic for F. hepatica surveillance in red deer, and its application here has revealed considerable geographic, temporal, sex and age related differences in F. hepatica prevalence in wild Scottish Highland red deer.