A defect in homologous recombination leads to increased translesion synthesisin E. coli

Scaling aquatic ecosystem processes like nutrient removal is critical for assessing the importance of streams and rivers to watershed nutrient export. We used pulse NH4+ enrichment experiments and measured net NH4+ uptake in 7 streams throughout a mountainous tropical river network in Puerto Rico to assess spatial variability in NH4+ uptake and to infer the physical, chemical, and biological characteristics that most influence its variation. Across 14 experiments, NH4+ uptake velocity (vf) ranged from 0.3 to 8.5 (mean = 2.7) mm/min and was positively related to algal biomass standing stock, measured as chlorophyll a. On average, 49% of experimentally added NH4+ was immediately transformed to NO3−, suggesting that nitrification can rival microbial and algal assimilation as a fate of streamwater NH4+. We considered the implications of our empirical results at the river-network scale based on a simple mass-balance model parameterized for the Río Mameyes watershed. Most catchment NH4+ inputs are delivered to 1st-order streams. Therefore, model results indicated that high NH4+ uptake rates in headwater streams limit NH4+ inputs to downstream reaches, thereby decreasing the role of larger streams in NH4+ removal at the river-network scale. In-stream nitrification resulted in additional NO3− inputs, which were more likely than NH4+ to be transported downstream because of lower biological demand for NO3− relative to NH4+. Given our estimates of catchment N loading to streams and rivers, we estimated that 39% of modeled watershed NO3− export was produced within the river network by nitrification. Together, these results suggest that streams and rivers can significantly transform the N load from their catchments

Variation in Detrital Resource Stoichiometry Signals Differential Carbon to Nutrient Limitation for Stream Consumers Across Biomes

Stoichiometric ratios of resources and consumers have been used to predict nutrient limitation across diverse terrestrial and aquatic ecosystems. In forested headwater streams, coarse and fine benthic organic matter (CBOM, FBOM) are primary basal resources for the food web, and the distribution and quality of these organic matter resources may therefore influence patterns of secondary production and nutrient cycling within stream networks or among biomes. We measured carbon (C), nitrogen (N), and phosphorus (P) content of CBOM and FBOM and calculated their stoichiometric ratios (C/N, C/P, N/P) from first- to fourth-order streams from tropical montane, temperate deciduous, and boreal forests, and tallgrass prairie, to compare the magnitude and variability of these resource types among biomes. We then used the ratios to predict nutritional limitations for consumers of each resource type. Across biomes, CBOM had consistently higher %C and %N, and higher and more variable C/N and C/P than FBOM, suggesting that microbial processing results in more tightly constrained elemental composition in FBOM than in CBOM. Biome-specific differences were observed in %P and N/P between the two resource pools; CBOM was lower in %P but higher in N/P than FBOM in the tropical montane and temperate deciduous forest biomes, while CBOM was higher in %P but similar in N/P than FBOM in the grassland and boreal forest biomes. Stable C-13 isotopes suggest that FBOM likely derives from CBOM in tropical and temperate deciduous forest, but that additional non-detrital components may contribute to FBOM in boreal forests and grasslands. Comparisons of stoichiometric ratios of CBOM and FBOM to estimated needs of aquatic detritivores suggest that shredders feeding on CBOM are more likely to experience nutrient (N and/or P) than C limitation, whereas collector-gatherers consuming FBOM are more likely to experience C than N and/or P limitation. Our results suggest that differences in basal resource elemental content and stoichiometric ratios have the potential to affect consumer production and ecosystem rates of C, N, and P cycling in relatively consistent ways across diverse biomes

Baseflow physical characteristics differ at multiple spatial scales in stream networks across diverse biomes

Context: Spatial scaling of ecological processes is facilitated by quantifying underlying habitat attributes. Physical and ecological patterns are often measured at disparate spatial scales limiting our ability to quantify ecological processes at broader spatial scales using physical attributes. Objective: We characterized variation of physical stream attributes during periods of high biological activity (i.e., baseflow) to match physical and ecological measurements and to identify the spatial scales exhibiting and predicting heterogeneity. Methods: We measured canopy cover, wetted width, water depth, and sediment size along transects of 1st–5th order reaches in five stream networks located in biomes from tropical forest to arctic tundra. We used hierarchical analysis of variance with three nested scales (watersheds, stream orders, reaches) to identify scales exhibiting significant heterogeneity in attributes and regression analyses to characterize gradients within and across stream networks. Results: Heterogeneity was evident at one or multiple spatial scales: canopy cover and water depth varied significantly at all three spatial scales while wetted width varied at two scales (stream order and reach) and sediment size remained largely unexplained. Similarly, prediction by drainage area depended on the attribute considered: depending on the watershed, increases in wetted width and water depth with drainage area were best fit with a linear, logarithmic, or power function. Variation in sediment size was independent of drainage area. Conclusions: The scaling of ecologically relevant baseflow physical characteristics will require study beyond the traditional bankfull geomorphology since predictions of baseflow physical attributes by drainage area were not always best explained by geomorphic power laws

Continental-scale decrease in net primary productivity in streams due to climate warming

Streams play a key role in the global carbon cycle. The balance between carbon intake through photosynthesis and carbon release via respiration influences carbon emissions from streams and depends on temperature. However, the lack of a comprehensive analysis of the temperature sensitivity of the metabolic balance in inland waters across latitudes and local climate conditions hinders an accurate projection of carbon emissions in a warmer future. Here, we use a model of diel dissolved oxygen dynamics, combined with high-frequency measurements of dissolved oxygen, light and temperature, to estimate the temperature sensitivities of gross primary production and ecosystem respiration in streams across six biomes, from the tropics to the arctic tundra. We find that the change in metabolic balance, that is, the ratio of gross primary production to ecosystem respiration, is a function of stream temperature and current metabolic balance. Applying this relationship to the global compilation of stream metabolism data, we find that a 1 °C increase in stream temperature leads to a convergence of metabolic balance and to a 23.6% overall decline in net ecosystem productivity across the streams studied. We suggest that if the relationship holds for similarly sized streams around the globe, the warming-induced shifts in metabolic balance will result in an increase of 0.0194 Pg carbon emitted from such streams every year

Phosphomimetic Substitution of Heterogeneous Nuclear Ribonucleoprotein A1 at Serine 199 Abolishes AKT-dependent Internal Ribosome Entry Site-transacting Factor (ITAF) Function via Effects on Strand Annealing and Results in Mammalian Target of Rapamycin Complex 1 (mTORC1) Inhibitor Sensitivity*

The relative activity of the AKT kinase has been demonstrated to be a major determinant of sensitivity of tumor cells to mammalian target of rapamycin (mTOR) complex 1 inhibitors. Our previous studies have shown that the multifunctional RNA-binding protein heterogeneous nuclear ribonucleoprotein (hnRNP) A1 regulates a salvage pathway facilitating internal ribosome entry site (IRES)-dependent mRNA translation of critical cellular determinants in an AKT-dependent manner following mTOR inhibitor exposure. This pathway functions by stimulating IRES-dependent translation in cells with relatively quiescent AKT, resulting in resistance to rapamycin. However, the pathway is repressed in cells with elevated AKT activity, rendering them sensitive to rapamycin-induced G1 arrest as a result of the inhibition of global eIF-4E-mediated translation. AKT phosphorylation of hnRNP A1 at serine 199 has been demonstrated to inhibit IRES-mediated translation initiation. Here we describe a phosphomimetic mutant of hnRNP A1 (S199E) that is capable of binding both the cyclin D1 and c-MYC IRES RNAs in vitro but lacks nucleic acid annealing activity, resulting in inhibition of IRES function in dicistronic mRNA reporter assays. Utilizing cells in which AKT is conditionally active, we demonstrate that overexpression of this mutant renders quiescent AKT-containing cells sensitive to rapamycin in vitro and in xenografts. We also demonstrate that activated AKT is strongly correlated with elevated Ser(P)199-hnRNP A1 levels in a panel of 22 glioblastomas. These data demonstrate that the phosphorylation status of hnRNP A1 serine 199 regulates the AKT-dependent sensitivity of cells to rapamycin and functionally links IRES-transacting factor annealing activity to cellular responses to mTOR complex 1 inhibition

Template for the description of cell-based toxicological test methods to allow evaluation and regulatory use of the data

Only few cell-based test methods are described by Organisation for Economic Co-operation and Development (OECD) test guidelines or other regulatory references (e.g., the European Pharmacopoeia). The majority of toxicity tests still falls into the category of non-guideline methods. Data from these tests may nevertheless be used to support regulatory decisions or to guide strategies to assess compounds (e.g., drugs, agrochemicals) during research and development if they fulfill basic requirements concerning their relevance, reproducibility and predictivity. Only a method description of sufficient clarity and detail allows interpretation and use of the data. To guide regulators faced with increasing amounts of data from non-guideline studies, the OECD formulated Guidance Document 211 (GD211) on method documentation for the purpose of safety assessment. As GD211 is targeted mainly at regulators, it leaves scientists less familiar with regulation uncertain as to what level of detail is required and how individual questions should be answered. Moreover, little attention was given to the description of the test system (i.e., cell culture) and the steps leading to it being established in the guidance. To address these issues, an annotated toxicity test method template (ToxTemp) was developed (i) to fulfill all requirements of GD211, (ii) to guide the user concerning the types of answers and detail of information required, (iii) to include acceptance criteria for test elements, and (iv) to define the cells sufficiently and transparently. The fully annotated ToxTemp is provided here, together with reference to a database containing exemplary descriptions of more than 20 cell-based tests.Erratum to Template for the Description of Cell-Based Toxicological Test Methods to Allow Evaluation and Regulatory Use of the Data.</p