Manipulating the EphB4-ephrinB2 axis to reduce metastasis in HNSCC.

The EphB4-ephrinB2 signaling axis has been heavily implicated in metastasis across numerous cancer types. Our emerging understanding of the dichotomous roles that EphB4 and ephrinB2 play in head and neck squamous cell carcinoma (HNSCC) poses a significant challenge to rational drug design. We find that EphB4 knockdown in cancer cells enhances metastasis in preclinical HNSCC models by augmenting immunosuppressive cells like T regulatory cells (Tregs) within the tumor microenvironment. EphB4 inhibition in cancer cells also amplifies their ability to metastasize through increased expression of genes associated with epithelial mesenchymal transition and hallmark pathways of metastasis. In contrast, vascular ephrinB2 knockout coupled with radiation therapy (RT) enhances anti-tumor immunity, reduces Treg accumulation into the tumor, and decreases metastasis. Notably, targeting the EphB4-ephrinB2 signaling axis with the engineered EphB4 ligands EFNB2-Fc-His and Fc-TNYL-RAW-GS reduces local tumor growth and distant metastasis in a preclinical model of HNSCC. Our data suggest that targeted inhibition of vascular ephrinB2 while avoiding inhibition of EphB4 in cancer cells could be a promising strategy to mitigate HNSCC metastasis.

Sequential hydrotalcite precipitation, microbial sulfate reduction and in situ hydrogen sulfide removal for neutral mine drainage treatment.

This study proposed and examined a new process flowsheet for treating neutral mine drainage (NMD) from an open-pit gold mine. The process consisted of three sequential stages: (1) in situ hydrotalcite (HT) precipitation; (2) low-cost carbon substrate driven microbial sulfate reduction; and (3) ferrosol reactive barrier for removing biogenic dissolved hydrogen sulfide (H2S). For concept validation, laboratory-scale columns were established and operated for a 140-days period with key process performance parameters regularly measured. At the end, solids recovered from various depths of the ferrosol column were analysed for elemental composition and mineral phases. Prokaryotic microbial communities in various process locations were characterised using 16S rRNA gene sequencing. Results showed that the Stage 1 HT-treatment substantially removed a range of elements (As, B, Ba, Ca, F, Zn, Si, and U) in the NMD, but not nitrate or sulfate. The Stage 2 sulfate reducing bioreactor (SRB) packed with 70 % (v/v) Eucalyptus woodchip, 1 % (w/v) ground (<1 mm) dried Typha biomass, and 10 % (w/v) NMD-pond sediment facilitated complete nitrate removal and stable sulfate removal of ca. 50 % (50 g-SO4 m-3 d-1), with an average H2S generation rate of 10 g-H2S m-3d-1. The H2S-removal performance of the Stage 3 ferrosol column was compared with a synthetic amorphous Fe-oxyhydroxide-amended sand control column. Although both columns facilitated excellent (95-100 %) H2S removal, the control column only enabled a further ca. 10 % sulfate reduction, giving an overall sulfate removal of 56 %. In contrast, the ferrosol enabled an extra 99.9 % sulfate reduction in the SRB effluent, leading to a near complete sulfate removal. Overall, the process successfully eliminated a range of metal/metalloid contaminants, nitrate, sulfate (2500 mg-SO4 L-1 in the NMD to <10 mg-SO4 L-1 in the final effluent) and H2S (>95 % removal). Further optimisation is required to minimise release of ferrous iron from the ferrosol barrier into the final effluent.

Quantification of the variability and penetration of per- and poly-fluoroalkyl substances through a concrete pad.

Historical use of aqueous film forming foams (AFFF) containing per- and poly-fluoroalkyl substances (PFAS) for fire-fighting activities has contributed to widespread contamination of infrastructure which can represent an ongoing source of PFAS to the surrounding environment. A concrete fire training pad with historical use of Ansulite and Lightwater AFFF formulations had PFAS concentrations measured to quantify spatial variability of PFAS within the pad. Surface chips and whole cores of concrete through to the underlying aggregate base were collected over the 24 × 9 m concrete pad and depth profiles of PFAS concentrations in nine cores were analysed. PFOS and PFHxS dominated the PFAS for surface samples, along the depth profile of cores and in the underlying plastic and aggregate material, with substantial variability in the concentrations of PFAS in the samples. Although there was variability of individual PFAS along the depth profile, higher surface concentrations of PFAS generally followed the designed movement of water across the pad. Total oxidisable precursor (TOP) assessments of one core indicated additional PFAS were present along the entire length of the core. This study highlights concentrations of PFAS (up to low µg/kg) from historical use of AFFF can occur throughout concrete, with the variable concentrations throughout the profile.

CTLA4 mRNA is downregulated by miR-155 in regulatory T cells, and reduced blood CTLA4 levels are associated with poor prognosis in metastatic melanoma patients.

Cytotoxic T lymphocyte-associated antigen-4 (CTLA-4) is an immune checkpoint expressed in regulatory T (Treg) cells and activated T lymphocytes. Despite its potential as a treatment strategy for melanoma, CTLA-4 inhibition has limited efficacy. Using data from The Cancer Genome Atlas (TCGA) melanoma database and another dataset, we found that decreased CTLA4 mRNA was associated with a poorer prognosis in metastatic melanoma. To investigate further, we measured blood CTLA4 mRNA in 273 whole-blood samples from an Australian cohort and found that it was lower in metastatic melanoma than in healthy controls and associated with worse patient survival. We confirmed these findings using Cox proportional hazards model analysis and another cohort from the US. Fractionated blood analysis revealed that Treg cells were responsible for the downregulated CTLA4 in metastatic melanoma patients, which was confirmed by further analysis of published data showing downregulated CTLA-4 surface protein expression in Treg cells of metastatic melanoma compared to healthy donors. Mechanistically, we found that secretomes from human metastatic melanoma cells downregulate CTLA4 mRNA at the post-transcriptional level through miR-155 while upregulating FOXP3 expression in human Treg cells. Functionally, we demonstrated that CTLA4 expression inhibits the proliferation and suppressive function of human Treg cells. Finally, miR-155 was found to be upregulated in Treg cells from metastatic melanoma patients compared to healthy donors. Our study provides new insights into the underlying mechanisms of reduced CTLA4 expression observed in melanoma patients, demonstrating that post-transcriptional silencing of CTLA4 by miRNA-155 in Treg cells may play a critical role. Since CTLA-4 expression is downregulated in non-responder melanoma patients to anti-PD-1 immunotherapy, targeting miRNA-155 or other factors involved in regulating CTLA4 expression in Treg cells without affecting T cells could be a potential strategy to improve the efficacy of immunotherapy in melanoma. Further research is needed to understand the molecular mechanisms regulating CTLA4 expression in Treg cells and identify potential therapeutic targets for enhancing immune-based therapies.

Treatment with anti-inflammatory viral serpin modulates immuno-thrombotic responses and improves outcomes in SARS-CoV-2 infected mice

1.Severe acute respiratory distress syndrome (ARDS) during SARS-CoV-2 (severe acute respiratory syndrome coronavirus-2) infection, manifests as uncontrolled lung inflammation and systemic thrombosis with high mortality. Anti-viral drugs and monoclonal antibodies can reduce COVID-19 severity if administered in the early viremic phase, but treatments for later stage immuno-thrombotic syndrome and long COVID are limited. Serine protease inhibitors (SERPINS) regulate activated proteases during thrombotic, thrombolytic and immune responses. The myxoma poxvirus-derived Serp-1 protein is a secreted immunomodulatory serpin that targets activated coagulation and complement protease pathways as part of a self-defense strategy to combat viral clearance by the innate immune system. When purified and utilized as an anti-immune therapeutic, Serp-1 is effective as an anti-inflammatory drug in multiple animal models of inflammatory lung disease and vasculitis. Here, we describe systemic treatment with purified PEGylated Serp-1 (PEGSerp-1) as a therapy for immuno-thrombotic complications during ARDS. Treatment with PEGSerp-1 in two distinct mouse-adapted SARS-CoV-2 models in C57Bl/6 and BALB/c mice reduced lung and heart inflammation, with improved clinical outcomes. PEGSerp-1 significantly reduced M1 macrophage invasion in the lung and heart by modifying urokinase-type plasminogen activator receptor (uPAR) and complement membrane attack complex (MAC). Sequential changes in urokinase-type plasminogen activator receptor (uPAR) and serpin gene expression were observed in lung and heart with PEGSerp-1 treatment. PEGSerp-1 is a highly effective immune-modulator with therapeutic potential for treatment of severe viral ARDS with additional potential to reduce late SARS-CoV-2 complications related to immune-thrombotic events that persist during long COVID. SignificanceSevere acute respiratory distress syndrome (ARDS) in SARS-CoV-2 infection manifests as uncontrolled tissue inflammation and systemic thrombosis with high mortality. Anti-viral drugs and monoclonal antibodies reduce COVID-19 severity if administered early, but treatments for later stage immuno-thrombosis are limited. Serine protease inhibitors (SERPINS) regulate thrombotic, thrombolytic and complement pathways. We investigate here systemic treatment with purified poxvirus-derived PEGSerp-1 as a therapeutic for immuno-thrombotic complications in viral ARDS. PEGSerp-1 treatment in two mouse-adapted SARS-CoV-2 models (C57Bl/6 and BALB/c) significantly reduced lung and heart inflammation and improved clinical outcomes, with sequential changes in thrombolytic (uPAR) and complement expression. PEGSerp-1 is a highly effective immune-modulator with therapeutic potential for immune-thrombotic complications in severe viral ARDS and has potential benefit for long COVID.

Expression of IL-37 Induces a Regulatory T-Cell-like Phenotype and Function in Jurkat Cells.

The anti-inflammatory cytokine interleukin-37 (IL-37) plays a key role in inhibiting innate and adaptive immunity. Past results have shown that IL-37 is elevated in human Treg cells compared to other T cell subsets and contributes to enhancing the Treg transcription factor, forkhead box protein P3 (FOXP3). However, it is unknown if ectopic expression of IL-37 in non-Treg CD4+ T cells can lead to the development of Treg phenotype and function. In the present study, we used a PrimeFlow® RNA assay and confirmed elevated IL37 expression in human Treg cells. We then stably transfected the non-Treg CD4+ T cell leukemia cell line, E6 Jurkat cells, with IL37 and found significant induction of the Treg phenotype. These IL-37-expressing Jurkat cells had elevated CTLA-4 and FOXP3 and produced IL-10. In conjunction with the Treg phenotype, IL-37-expressing Jurkat cells suppressed T cell activation/proliferation, comparable to human primary Treg cells. The creation of this stable human Treg-like cell line has the potential to provide further assistance for in vitro studies of human Treg cells, as it is more convenient than the use of primary human Treg cells. Furthermore, it provides insights into Treg cell biology and function.

Correction: Method for extraction and analysis of per- and poly-fluoroalkyl substances in contaminated asphalt.

Correction for 'Method for extraction and analysis of per- and poly-fluoroalkyl substances in contaminated asphalt' by Prashant Srivastava et al., Anal. Methods, 2022, 14, 1678-1689, https://doi.org/10.1039/D2AY00221C.

Volatilization Potential of Per- and Poly-fluoroalkyl Substances from Airfield Pavements and during Recycling of Asphalt.

Per- and poly-fluoroalkyl substances (PFAS) in water are typically present in their ionic (nonvolatile) forms; however, these can transition to their nonionic (volatile) forms when in contact with organic solvents and organic matrices. In particular, when PFAS are dissolved in organic solvents such as residues left from firefighting foams, fuels, and bitumen present in asphalt, the equilibrium between ionic and nonionic forms can trend toward more volatile nonionic forms of PFAS. We assessed the volatility of common PFAS based on calculated and available experimental data across ambient temperature ranges experienced by airfield pavements and at elevated temperatures associated with reworking asphalts for reuse. Volatilities are shown to be comparable to hydrocarbons in the semivolatile range, suggesting that volatilization is a viable loss mechanism for some PFAS that are nonvolatile in water. The present study points to future investigative needs for this unexplored mass loss mechanism and potential exposure pathway. Environ Toxicol Chem 2022;41:2202-2208. © 2022 Commonwealth of Australia. Environmental Toxicology and Chemistry published by Wiley Periodicals LLC on behalf of SETAC.

Method for extraction and analysis of per- and poly-fluoroalkyl substances in contaminated asphalt.

The legacy use of aqueous film-forming foam (AFFF) has led to the generation of large volumes of per- and poly-fluoroalkyl substances (PFAS)-contaminated asphalt materials, especially at airports and fire training areas. The management of such PFAS-contaminated asphalt materials requires an understanding of PFAS concentrations in these materials. This study, therefore, aimed to develop a suitable extraction methodology for the analysis of 22 target PFAS (i.e., carboxylic acids, sulfonic acids and fluorotelomers) in asphalt materials. A series of experiments was conducted to optimise extraction solvent composition, as well as to assess the performance of the chosen method under various conditions (i.e., sonication temperature, PFAS contamination level, asphalt core composition and timing of stable isotope addition used as internal standard). The methanol-based extractants performed best due to their accuracy and precision, which were within the acceptable range (extraction efficiency between 70 and 130% and RSD < 20%). The method which involved three successive extractions with methanol/1% NH3 by ultrasonication at 25 °C was selected due to its performance and ease of operation. The mean recovery of a vast majority of PFAS was found to be in the acceptable range. Tests on the timing of addition of stable isotope (SI)-labelled PFAS internal standards indicate that the recoveries obtained, regardless of when the stable isotopes were added, were within the acceptable range for PFAS. The accuracy and precision of PFAS recoveries were not affected by PFAS spike level (2 µg kg-1 and 200 µg kg-1), as well as sample composition (based on the location of asphalt material in the field). Low RSDs were achieved for asphalt cores collected from a contaminated site covering a wide range of concentrations (from LOQ to 2135 mg kg-1), demonstrating the suitability of the sample preparation method for real-world samples. The results from the interlaboratory testing were also in good agreement and validated the proposed PFAS extraction and analytical approach.

EGCG Inhibits Tumor Growth in Melanoma by Targeting JAK-STAT Signaling and Its Downstream PD-L1/PD-L2-PD1 Axis in Tumors and Enhancing Cytotoxic T-Cell Responses.

Over the last decade, therapies targeting immune checkpoints, such as programmed death-1 (PD-1), have revolutionized the field of cancer immunotherapy. However, low response rates and immune-related adverse events remain a major concern. Here, we report that epigallocatechin gallate (EGCG), the most abundant catechin in green tea, inhibits melanoma growth by modulating an immune response against tumors. In vitro experiments revealed that EGCG treatment inhibited interferon-gamma (IFN-γ)-induced PD-L1 and PD-L2 expression and JAK-STAT signaling. We confirmed that this effect was driven by inhibiting STAT1 gene expression and STAT1 phosphorylation, thereby downregulating the PD-L1/PD-L2 transcriptional regulator IRF1 in both human and mouse melanoma cells. Animal studies revealed that the in vivo tumor-inhibitory effect of EGCG was through CD8+ T cells and that the inhibitory effect of EGCG was comparable to anti-PD-1 therapy. However, their mechanisms of action were different. Dissimilar to anti-PD-1 treatment that blocks PD-1/PD-L1 interaction, EGCG inhibited JAK/STAT signaling and PD-L1 expression in tumor cells, leading to the re-activation of T cells. In summary, we demonstrate that EGCG enhances anti-tumor immune responses by inhibiting JAK-STAT signaling in melanoma. EGCG could be used as an alternative treatment strategy to target the PD-L1/PD-L2-PD-1 axis in cancers.

Treatment of neutral gold mine drainage by sequential in situ hydrotalcite precipitation, and microbial sulfate and cyanide removal.

This study proposed and validated a method integrating in situ hydrotalcite precipitation (Virtual Curtain™ (VC) technology) with bioprocess for treating a cyanide (CN)-augmented (ca. 5 mg-CN L-1) sulfate-laden neutral mine drainage, from a waste rock dump (WD2) of an Australian gold mine. Efficacies of various carbon (C) sources (ethanol, lactate, and two natural substrates; Eucalyptus wood sawdust (EW) and Typha biomass (TB)) for promoting microbial reduction in both: CN-augmented WD2 water and VC-treated CN-augmented WD2 water were assessed in a 60-days microcosms study at 30 °C. The microcosms were monitored over time for pH, redox potential, dissolved hydrogen sulfide, chloride, nitrite, nitrate, sulfate, phosphate, biogas production, dissolved organic carbon, total dissolved nitrogen, and dissolved CN. The VC treatment removed a range of metals (Mg, Ni and Zn) and metalloid Se from the CN-augmented WD2 water to below detection. Other elements substantially reduced in concentration included Ba, F, Si and U. However, the VC treatment did not remove substantial nitrate, sulfate or CN. Microcosm trials revealed that the indigenous microbial community in WD2 could effectively denitrify and reduce sulfate, with TB was the most efficient C source for promoting sulfate and CN removal; whereas, EW facilitated only marginally higher sulfate reduction compared with controls. The highest sulfate reduction rate (76 g-SO42- m-3 d-1) was achieved with VC-treated water amended with TB, indicating that VC pre-treatment was beneficial. Further, all treatments amended with external C, facilitated 100% removal of dissolved CN after 60 days, whereas only partial (65%) CN removal was recorded in the control. Overall, the proposed integrated method appears a viable option for treating neutral gold mine drainage.

Thermal flux, fugitive gas emission and geotechnical instability in a complex tailings legacy site.

Several years after decommissioning, a magnesium dross and mixed waste heap at a former industrial facility is still reactive, as evidenced by the emission of heat, Volatile Organic Carbon (VOCs), acetylene (C2H2), cyanide (HCN) and ammonia (NH3) from deep, discordant, epigenetic fissures. To evaluate the longer-term stability of the waste heap material, four cores were collected to evaluate vertical variations in temperature, moisture, gas composition, geochemistry, and mineralogy. Temperature increased with depth and peaked at around 8 m, reaching in excess of 90 °C. The waste heap was a mixture of unreacted materials (mainly MgO and CaO) and a variety of hydrated secondary reaction products. Formation of the latter could account for the generation of heat and creation of the fissures via thermal and secondary mineral volumetric expansion. With a large inventory of unreacted CaO and MgO and substantial in situ water present, the waste heap will probably remain reactive in the foreseeable future. Importantly, the CaO/MgO ratio of solid materials in the waste heap provides a useful proxy for down hole temperature, pH, and fugitive gas concentrations. Fugitive gases emitted by the waste heap are related to the reaction of co-existing minerals in the heap based on an historical analysis of site waste disposal. These waste materials include calcium carbide (CaC2), magnesium nitride (Mg3N2) and calcium cyanamide (CaCN2). Capping to limit the ingress of additional meteoric water and targeted venting to facilitate cooling and the controlled release and dispersion of gases are recommended to manage the environmental risk.

Contrasting effects and mode of dredging and in situ adsorbent amendment for the control of sediment internal phosphorus loading in eutrophic lakes.

Dredging and in situ adsorbent inactivation are two methods which are frequently used in eutrophic water bodies such as ponds, lakes and estuaries to control internal phosphorus (P) loading from sediments. However, their effects and modes on the control of sediment P loading has been seldom compared. In this study, a long-term sediment core incubation experiment in the field was undertaken to investigate changes in sediment P loading (P fluxes, supply ability and forms of P and transformation) comparing two remediation techniques, that of lanthanum-modified bentonite (LMB) addition or dredging to a control. A 360-day field investigation indicated that LMB addition more effectively reduced pore water P concentrations and sediment P fluxes than dredging in comparison with the control. On average, dredging and in situ LMB inactivation reduced the P flux by 82% and 90%, respectively relative to the control sediment. Whilst both the LMB inactivation and dredging can reduce the mobile P concentration, the impact of LMB in reducing mobile P was demonstrated to be more prolonged than that of dredging after 360 days. The P fraction composition in the LMB inactivated sediment differed significantly from the dredged and control sediment. Contrary to physical removal of dredging, chemical transformation of sediment mobile P and Al-P into Ca-P is the main function mode of LMB for sediment internal P control. Both LMB addition and dredging caused changes in the composition of sediment bacterial communities. Whilst LMB addition increased bacterial diversity, dredging temporarily reduced it. This study indicates that in situ inactivation by LMB is superior to dredging in the long-term control of sediment P loading.

Sequential hydrotalcite precipitation and biological sulfate reduction for acid mine drainage treatment.

Hydrotalcite precipitation is a promising technology for the on-site treatment of acid mine drainage (AMD). This technology is underpinned by the synthesis of hydrotalcite that can effectively remove various contaminants. However, hydrotalcite precipitation has only limited capacity to facilitate sulfate removal from AMD. Therefore, the feasibility of coupling biological sulfate reduction with the hydrotalcite precipitation to maximize sulfate removal was evaluated in this study. AMD emanating from a gold mine (pH 4.3, sulfate 2000 mg L-1, with various metals including Al, Cd, Co, Cu, Fe, Mn, Ni, Zn) was first treated using the hydrotalcite precipitation. Subsequently, biological treatment of the post-hydrotalcite precipitation effluent was conducted in an ethanol-fed fluidized bed reactor (FBR) at a hydraulic retention time (HRT) of 0.8-1.6 day. The hydrotalcite precipitation readily neutralized the acidity of AMD and removed 10% of sulfate and over 99% of Al, Cd, Co, Cu, Fe, Mn, Ni, Zn. The overall sulfate removal increased to 73% with subsequent FBR treatment. Based on 454 pyrosequencing of 16S rRNA genes, the identified genera of sulfate-reducing bacteria (SRB) included Desulfovibrio, Desulfomicrobium and Desulfococcus. This study showed that sulfate-rich AMD can be effectively treated by integrating hydrotalcite precipitation and a biological sulfate reducing FBR.

Soil carbon stocks under grazed pasture and pasture-tree systems.

There is increasing interest in the potential of trees to sequester carbon (C) in above- and below-ground stocks to mitigate against increasing concentrations of greenhouse gases (GHG). This study determined whether pasture-tree (PT) systems influence soil C stocks compared with open pasture (OP) by sampling four sites with trees aged 14 to16 years. Poplars (Populus spp.) at Tikokino and Woodville and alders (Alnus spp.) at Poukawa and Ruakura were planted on contrasting soils (Haplustands, Endoaquepts, Durustalfs and Humaquepts, respectively). Trees at all four sites were arranged in partial-Nelder radial planting designs, with five stem densities ranging from 67 to 1276 stems ha-1. Soils were sampled at five stem density classes, along with adjacent OP areas in the same paddock, to a depth of 1 m (0-75, 75-150, 150-300, 300-600, 600-1000 mm). At three of the four sites, root mass density was greater (P < 0.05) in PT than in OP systems. At Woodville, estimates of total soil C mass to 1 m tended to be greater (P = 0.08) in the OP than in the PT system (200 vs. 163 Mg C ha-1, respectively), whereas no significant differences in total soil C masses between OP and PT were shown at the remaining sites (P > 0.10). Despite the limited statistical significance, estimates of total soil C mass at Tikokino and Woodville (sites with poplars) were 11 and 18% greater in OP than in PT systems, whereas estimates at Poukawa and Ruakura (sites with alders) were 2 and 6% greater in PT than in OP systems. Under the current conditions, our study suggests that tree species may be an additional factor influencing the C cycle and C accumulation in soils and need to be considered in the building of our soil C inventories.

Lanthanum modified bentonite behaviour and efficiency in adsorbing phosphate in saline waters.

Lanthanum-modified bentonite (LMB, commercially called Phoslock®) has been widely applied in freshwater systems to manage eutrophication. Little is known, however, about its behaviour and efficiency in binding filterable reactive phosphorus (FRP) in saline environments. We assessed if LMB would adsorb phosphate over a range of salinities (0-32 ppth) comparing the behaviour in seawater salts and equivalent concentrations of NaCl. Lanthanum release from the bentonite matrix was measured and the La species prevailing in saline environments were evaluated through chemical equilibrium modelling. We demonstrated that LMB was able to adsorb FRP in all the salinities tested. Filterable lanthanum (FLa) concentrations were similarly low (<5 µgL-1) at all seawater salinities but considerably elevated, on occasion >2000 times greater in equivalent NaCl salinities. Mineralogical analysis indicates that La present in the clay interlayer was (partially) replaced by Na/Ca/Mg present in the seawater and a possible secondary P-reactive phase was formed, such as kozoite (LaCO3OH) or lanthanite (La2(CO3)3·8H2O) that may be physically dissociated from the LMB. Geochemical modelling also indicates that most FLa dissociated from LMB would be precipitated as a carbonate complex. In light of the identification of reactive intermediate phases, further studies including ecotoxicologial assays are required to assess any deleterious effects from the application of LMB to saline waters.

Lanthanum in Water, Sediment, Macrophytes and chironomid larvae following application of Lanthanum modified bentonite to lake Rauwbraken (The Netherlands).

Lanthanum Modified Bentonite (LMB; Phoslock®) is used to mitigate eutrophication by binding phosphate released from sediments. This study investigated the fate of lanthanum (La) from LMB in water, sediment, macrophytes, and chironomid larvae in Lake Rauwbraken (The Netherlands). Before the LMB application, water column filterable La (FLa) was 0.02 µg L-1, total La (TLa) was 0.22 µg L-1. In sediment the total La ranged 0.03-1.86 g m-2. The day after the application the maximum FLa concentration in the water column was 44 µg L-1, TLa was 528 µg L-1, exceeding the Dutch Maximum Permissible Concentrations (MPC) of 10.1 µg L-1 by three to fourfold. TLa declined below the MPC after 15 days, FLa after 75 days. After ten years, FLa was 0.4 µg L-1 and TLa was 0.7 µg L-1. Over the post-application years, FLa and TLa showed statistically significant downward trends. While the LMB settled homogeneously on sediment, after 3 years it redistributed to 0.2-5.4 g La m-2 within shallow zones, and 30.7 g m-2 to 40.0 g La m-2 in deeper zones. In the upper 20 cm of sediment, La concentrations were 7-6702 mg kg -1 dry weight (DW) compared to 0.5-7.0 mg kg-1 before application. Pre-application anaerobic sediment release of FLa was 0.006 mg m-2 day-1. Three months after the application it was 1.02 mg m-2 day-1. Three years later it was 0.063 mg m-2 day-1. Before application La in plants was 0.8-5.1 mg La kg-1 DW, post-application values were up to 2925 mg La kg-1 DW. In chironomid larvae, La increased from 1.7 µg g-1 DW before application to 1421 µg g-1 DW after one month, 3 years later it was 277 µg g-1 DW. Filtration experiments indicate FLa is not truly dissolved free La3+ cations.

Engineering and kinetic aspects of bacterial uranium reduction for the remediation of uranium contaminated environments.

Biological reduction of soluble uranium from U(VI) to insoluble U(IV) coupled to the oxidation of an electron donor (hydrogen or organic compounds) is a potentially cost-efficient way to reduce the U concentrations in contaminated waters to below regulatory limits. A variety of microorganisms originating from both U contaminated and non-contaminated environments have demonstrated U(VI) reduction capacity under anaerobic conditions. Bioreduction of U(VI) is considered especially promising for in situ remediation, where the activity of indigenous microorganisms is stimulated by supplying a suitable electron donor to the subsurface to contain U contamination to a specific location in a sparingly soluble form. Less studied microbial biofilm-based bioreactors and bioelectrochemical systems have also shown potential for efficient U(VI) reduction to remove U from contaminated water streams. This review compares the advantages and challenges of U(VI)-reducing in situ remediation processes, bioreactors and bioelectrochemical systems. In addition, the current knowledge of U(VI) bioreduction mechanisms and factors affecting U(VI) reduction kinetics (e.g. pH, temperature, and the chemical composition of the contaminated water) are discussed, as both of these aspects are important in designing efficient remediation processes.