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The oxidation of sulfide minerals such as pyrite present in waste rock results in elevated sulfate, enhanced metal loadings and in many cases low pH conditions. Recently, many mines have opened in remote areas, including regions subject to permafrost conditions. In these regions, freeze-thaw cycles and the possible development of permafrost in mine waste add to the complexity of weathering processes, drainage volumes and mass loadings. To assess weathering in these waste rock piles, the reactive transport code MIN3P-HPC has been enhanced by implementing constitutive relationships related to freeze-thaw cycles that control flow patterns, solute transport, generation and transport of heat, as well as geochemical reactions and their rates. Simulations of a hypothetical pyrite-rich waste rock pile placed onto natural permafrost were conducted under reference climate conditions. Additionally, the effect of a warming climate was also studied through a sensitivity analysis. The simulation results indicate a potentially strong coupled effect of sulfide mineral weathering rates and a warming climate on the evolution and persistence of permafrost within waste rock piles and the release of acidic drainage. For relatively low sulfide mineral oxidation rates, the simulations indicate that permafrost can develop within waste rock piles, even under warming climate conditions. However, the results for low reactivity also show that mass loadings can increase by >50% in response to a slight warming of climate (3°C), relative to reference climate conditions. For the chosen reference reaction rates, permafrost develops under reference climate conditions in the simulated waste rock pile; however, permafrost cannot be maintained for a marginally warmer climate, leading to internal heating of the pile and substantially increased production of acidic drainage (>550%). For high reaction rates, the simulations suggest that internal heating takes place irrespective of climate conditions. Evaluation of thermal covers indicates that significant reductions of mass loadings can be achieved for piles with low and reference reactivity (91–99% in comparison to uncovered piles), but also suggest that thermal covers can be ineffective for piles with high sulfide content and reactivity. Together, these simulations provide insights into the complex interactions controlling waste rock weathering in cold-region climates.

Death receptor signaling is critical for cell death, inflammation, and immune homeostasis. Hijacking death receptors and their corresponding adaptors through type III secretion system (T3SS) effectors has been evolved to be a bacterial evasion strategy. NleB from enteropathogenic Escherichia coli (EPEC) and SseK1/2/3 from Salmonella enterica serovar Typhimurium (S. Typhimurium) can modify some death domain (DD) proteins through arginine-GlcNAcylation. Here, we performed a substrate screen on 12 host DD proteins with conserved arginine during EPEC and Salmonella infection. NleB from EPEC hijacked death receptor signaling through tumor necrosis factor receptor 1 (TNFR1)-associated death domain protein (TRADD), FAS-associated death domain protein (FADD), and receptor-interacting serine/threonine-protein kinase 1 (RIPK1), whereas SseK1 and SseK3 disturbed TNF signaling through the modification of TRADD Arg235/Arg245 and TNFR1 Arg376, respectively. Furthermore, mouse infection studies showed that SseK1 but not SseK3 rescued the bacterial colonization deficiency contributed by the deletion of NleBc (Citrobacter NleB), indicating that TRADD was the in vivo substrate. The result provides an insight into the mechanism by which attaching and effacing (A/E) pathogen manipulate TRADD-mediated signaling and evade host immune defense through T3SS effectors.

Many Gram-negative bacterial pathogens utilize the type III secretion system (T3SS) to inject virulence factors, named effectors, into host cells. These T3SS effectors manipulate host cellular signaling pathways to facilitate bacterial pathogenesis. Death receptor signaling plays an important role in eukaryotic cell death pathways. NleB from enteropathogenic Escherichia coli (EPEC) and SseK1/3 from Salmonella enterica serovar Typhimurium (S. Typhimurium) are T3SS effectors. They are defined as a family of arginine GlcNAc transferase to modify a conserved arginine residue in the death domain (DD) of the death receptor TNFR and their corresponding adaptors to hijack death receptor signaling. Here we identified that these enzymes, NleB, SseK1, and SseK3 could catalyze auto-GlcNAcylation. Residues, including Arg13/53/159/293 in NleB, Arg30/158/339 in SseK1, and Arg153/184/305/335 in SseK3 were identified as the auto-GlcNAcylation sites by mass spectrometry. Mutation of the auto-modification sites of NleB, SseK1, and SseK3 abolished or attenuated the capability of enzyme activity toward their death domain targets during infection. Loss of this ability led to the increased susceptibility of the cells to TNF- or TRAIL-induced cell death during bacterial infection. Overall, our study reveals that the auto-GlcNAcylation of NleB, SseK1, and SseK3 is crucial for their biological activity during infection.

In order to reduce contaminant mass loadings, thermal cover systems may be incorporated in the design of waste rock piles located in regions of continuous permafrost. In this study, reactive transport modeling was used to improve the understanding of coupled thermo-hydrological and chemical processes controlling the evolution of a covered waste rock pile located in Northern Canada. Material properties from previous field and laboratory tests were incorporated into the model to constrain the simulations. Good agreement between simulated and observational temperature data indicates that the model is capable of capturing the coupled thermo-hydrological processes occurring within the pile. Simulations were also useful for forecasting the pile’s long-term evolution with an emphasis on water flow and heat transport mechanisms, but also including geochemical weathering processes and sulfate mass loadings as an indicator for the release of contaminated drainage. An uncertainty analysis was carried out to address different scenarios of the cover’s performance as a function of the applied infiltration rate, accounting for the impacts of evaporation, runoff, and snow ablation. The model results indicate that the cover performance is insensitive to the magnitude of recharge rates, except for limited changes of the flow regime in the shallow active layer. The model was expanded by performing an additional sensitivity analysis to assess the role of cover thicknesses. The simulated results reveal that a cover design with an appropriate thickness can effectively minimize mass loadings in drainage by maintaining the active layer completely within the cover.

The NLRP1 inflammasome functions as canonical cytosolic sensor in response to intracellular infections and is implicated in auto-inflammatory diseases. But the regulation and signal transduction mechanisms of NLRP1 are incompletely understood. Here, we show that the T60 variant of CARD8, but not the canonical T48 isoform, negatively regulates the NLRP1 inflammasome activation by directly interacting with the receptor molecule NLRP1 and inhibiting inflammasome assembly. Furthermore, our results suggest that different ASC preference in three types of inflammasomes, namely the ASC-indispensable NLRP1 inflammasome, ASC-dispensable mNLRP1b inflammasome and ASC-independent CARD8 inflammasome, is mainly caused by the CARD domain, not the UPA subdomain. Based on the systematic site-directed mutagenesis and structural analysis, we find that signal transduction of the NLRP1 inflammasome relies on multiple interaction surfaces at its CARD domain. Finally, our results partly explain how mutations in NLRP1 lead to its constitutive activation in auto-inflammatory diseases. In conclusion, our study not only reveals how CARD8 downregulates the NLRP1 inflammasome activation, but also provides insights into the assembly mechanisms of CARD-containing inflammasomes.

Conserved regions of the antibody molecule impact its downstream biological effects. Here the authors show that a rigid hinge conformation increases the agonistic activity of CD40 and DR5 antibodies, distinctly from FcγR-binding, suggesting that the hinge and FcR binding regions can be separately modified to optimize therapies.