Geochemistry of mine pool discharges in the Pittsburgh coal basin

Seventy-two mine water discharges from flooded underground mines of the Pittsburgh coal bed were sampled for chemistry during 2012. Statistical techniques including correlation, principal component analysis (PCA), and cluster analysis (CA) were utilized to evaluate geochemical relationships. The distributions of most solute concentrations are non-parametric. Strong correlations were observed between constituents of pyrite and carbonate dissolution (Fe, SO 4, Ca, Mg) as well as Na, Sr, and K. Several waters with high scores on PC1 are moderately saline with average concentrations of Na (1199 mg/L), Cl (350 mg/L), Br (1.69 mg/L), and Sr (4.13 mg/L). Waters with highly negative scores on PC2 are acidic with maximum concentrations of Fe3+ (35.3mg/L) and Al (23.3 mg/L) and discharge from shallow mines along the eastern margin of the coal subcrop. Waters with highly positive scores on PC2 are alkaline waters from deep, fully flooded mines with pH (6.3) and alkalinity (876 mg/L). PC3 has high scores primarily due to elevated Ba. Pittsburgh coal bed waters display, on average, SO4 (923 mg/L), Na (267 mg/L), alkalinity (226 mg/L as CaCO3), Ca (245 mg/L), Cl (100 mg/L), and Fe (45 mg/L) with traces of brine constituents. Diagnostic variations in concentrations of Ba, Br, Sr, Cl, and Al reveal spatial trends and partitioning into six clusters

Computational Insights on the Competing Effects of Nitric Oxide in Regulating Apoptosis

Despite the establishment of the important role of nitric oxide (NO) on apoptosis, a molecular- level understanding of the origin of its dichotomous pro- and anti-apoptotic effects has been elusive. We propose a new mathematical model for simulating the effects of nitric oxide (NO) on apoptosis. The new model integrates mitochondria-dependent apoptotic pathways with NO-related reactions, to gain insights into the regulatory effect of the reactive NO species N2O3, non-heme iron nitrosyl species (FeLnNO), and peroxynitrite (ONOO−). The biochemical pathways of apoptosis coupled with NO-related reactions are described by ordinary differential equations using mass-action kinetics. In the absence of NO, the model predicts either cell survival or apoptosis (a bistable behavior) with shifts in the onset time of apoptotic response depending on the strength of extracellular stimuli. Computations demonstrate that the relative concentrations of anti- and pro-apoptotic reactive NO species, and their interplay with glutathione, determine the net anti- or pro-apoptotic effects at long time points. Interestingly, transient effects on apoptosis are also observed in these simulations, the duration of which may reach up to hours, despite the eventual convergence to an anti-apoptotic state. Our computations point to the importance of precise timing of NO production and external stimulation in determining the eventual pro- or anti-apoptotic role of NO

Animal welfare testing for shooting and darting free-ranging wildlife: a review and recommendations

Several important techniques for managing wildlife rely on ballistics (the behaviour of projectiles), including killing techniques (shooting) as well as capture and marking methods (darting). Because all ballistic techniques have the capacity to harm animals, animal welfare is an important consideration. Standardised testing approaches that have allowed refinement for other physical killing and capture methods (e.g. traps for mammals) have not been applied broadly to ballistic methods. At the same time, new technology is becoming available for shooting (e.g. subsonic and lead-free ammunition) and darting (e.g. dye-marker darts). We present several case studies demonstrating (a) how basic ballistic testing can be performed for novel firearms and/or projectiles, (b) the benefits of identifying methods producing undesirable results before operational use, and (c) the welfare risks associated with bypassing testing of a technique before broad-scale application. Following the approach that has been used internationally to test kill-traps, we suggest the following four-step testing process: (1) range and field testing to confirm accuracy and precision, the delivery of appropriate kinetic energy levels and projectile behaviour, (2) post-mortem assessment of ballistic injury in cadavers, (3) small-scale live animal pilot studies with predetermined threshold pass/fail levels, and (4) broad-scale use with reporting of the frequency of adverse animal welfare outcomes. Wepresent this as a practical approach for maintaining and improving animal welfare standards when considering the use of ballistic technology for wildlife management

Oxidation state-specific fluorescent copper sensors reveal oncogene-driven redox changes that regulate labile copper(II) pools

Copper is an essential metal nutrient for life that often relies on redox cycling between Cu(I) and Cu(II) oxidation states to fulfill its physiological roles, but alterations in cellular redox status can lead to imbalances in copper homeostasis that contribute to cancer and other metalloplasias with metal-dependent disease vulnerabilities. Copper-responsive fluorescent probes offer powerful tools to study labile copper pools, but most of these reagents target Cu(I), with limited methods for monitoring Cu(II) owing to its potent fluorescence quenching properties. Here, we report an activity-based sensing strategy for turn-on, oxidation state-specific detection of Cu(II) through metal-directed acyl imidazole chemistry. Cu(II) binding to a metal and oxidation state-specific receptor that accommodates the harder Lewis acidity of Cu(II) relative to Cu(I) activates the pendant dye for reaction with proximal biological nucleophiles and concomitant metal ion release, thus avoiding fluorescence quenching. Copper-directed acyl imidazole 649 for Cu(II) (CD649.2) provides foundational information on the existence and regulation of labile Cu(II) pools, including identifying divalent metal transporter 1 (DMT1) as a Cu(II) importer, labile Cu(II) increases in response to oxidative stress induced by depleting total glutathione levels, and reciprocal increases in labile Cu(II) accompanied by decreases in labile Cu(I) induced by oncogenic mutations that promote oxidative stress

Animal welfare testing for shooting and darting free-ranging wildlife: a review and recommendations

Several important techniques for managing wildlife rely on ballistics (the behaviour of projectiles), including killing techniques (shooting) as well as capture and marking methods (darting). Because all ballistic techniques have the capacity to harm animals, animal welfare is an important consideration. Standardised testing approaches that have allowed refinement for other physical killing and capture methods (e.g. traps for mammals) have not been applied broadly to ballistic methods. At the same time, new technology is becoming available for shooting (e.g. subsonic and lead-free ammunition) and darting (e.g. dye-marker darts). We present several case studies demonstrating (a) how basic ballistic testing can be performed for novel firearms and/or projectiles, (b) the benefits of identifying methods producing undesirable results before operational use, and (c) the welfare risks associated with bypassing testing of a technique before broad-scale application. Following the approach that has been used internationally to test kill-traps, we suggest the following four-step testing process: (1) range and field testing to confirm accuracy and precision, the delivery of appropriate kinetic energy levels and projectile behaviour, (2) post-mortem assessment of ballistic injury in cadavers, (3) small-scale live animal pilot studies with predetermined threshold pass/fail levels, and (4) broad-scale use with reporting of the frequency of adverse animal welfare outcomes. We present this as a practical approach for maintaining and improving animal welfare standards when considering the use of ballistic technology for wildlife management. ethics, fertility control, human dimensions, pest control, population control

Oncogenic KRAS Induces NIX-Mediated Mitophagy to Promote Pancreatic Cancer.

Activating KRAS mutations are found in nearly all cases of pancreatic ductal adenocarcinoma (PDAC), yet effective clinical targeting of oncogenic KRAS remains elusive. Understanding of KRAS-dependent PDAC-promoting pathways could lead to the identification of vulnerabilities and the development of new treatments. We show that oncogenic KRAS induces BNIP3L/NIX expression and a selective mitophagy program that restricts glucose flux to the mitochondria and enhances redox capacity. Loss of Nix restores functional mitochondria to cells, increasing demands for NADPH reducing power and decreasing proliferation in glucose-limited conditions. Nix deletion markedly delays progression of pancreatic cancer and improves survival in a murine (KPC) model of PDAC. Although conditional Nix ablation in vivo initially results in the accumulation of mitochondria, mitochondrial content eventually normalizes via increased mitochondrial clearance programs, and pancreatic intraepithelial neoplasia (PanIN) lesions progress to PDAC. We identify the KRAS-NIX mitophagy program as a novel driver of glycolysis, redox robustness, and disease progression in PDAC. SIGNIFICANCE: NIX-mediated mitophagy is a new oncogenic KRAS effector pathway that suppresses functional mitochondrial content to stimulate cell proliferation and augment redox homeostasis. This pathway promotes the progression of PanIN to PDAC and represents a new dependency in pancreatic cancer.This article is highlighted in the In This Issue feature, p. 1143.Includes: M.P. Murphy is supported by Medical Research Council UK (MC_UU_00015/3) and by a Wellcome Trust Investigator award (110159/Z/15/Z). T.J. Humpton and K.H. Vousden are supported by Cancer Research UK and by ERC grant 322842-METABOp53. T.J. Humpton was also supported by the Gates Cambridge Trust. G.I. Evan and D. Lu are supported by Cancer Research UK Programme Grant A12077 (principal investigator: G.I. Evan)

Oncogene-induced Nrf2 transcription promotes ROS detoxification and tumorigenesis

Reactive oxygen species (ROS) are mutagenic and may thereby promote cancer. Normally, ROS levels are tightly controlled by an inducible antioxidant program that responds to cellular stressors and is predominantly regulated by the transcription factor Nrf2 (also known as Nfe2l2) and its repressor protein Keap1 (refs 2-5). In contrast to the acute physiological regulation of Nrf2, in neoplasia there is evidence for increased basal activation of Nrf2. Indeed, somatic mutations that disrupt the Nrf2-Keap1 interaction to stabilize Nrf2 and increase the constitutive transcription of Nrf2 target genes were recently identified, indicating that enhanced ROS detoxification and additional Nrf2 functions may in fact be pro-tumorigenic. Here, we investigated ROS metabolism in primary murine cells following the expression of endogenous oncogenic alleles of Kras, Braf and Myc, and found that ROS are actively suppressed by these oncogenes. K-Ras(G12D), B-Raf(V619E) and Myc(ERT2) each increased the transcription of Nrf2 to stably elevate the basal Nrf2 antioxidant program and thereby lower intracellular ROS and confer a more reduced intracellular environment. Oncogene-directed increased expression of Nrf2 is a new mechanism for the activation of the Nrf2 antioxidant program, and is evident in primary cells and tissues of mice expressing K-Ras(G12D) and B-Raf(V619E), and in human pancreatic cancer. Furthermore, genetic targeting of the Nrf2 pathway impairs K-Ras(G12D)-induced proliferation and tumorigenesis in vivo. Thus, the Nrf2 antioxidant and cellular detoxification program represents a previously unappreciated mediator of oncogenesis