US climate policy yields water quality cobenefits in the Mississippi Basin and Gulf of Mexico.

We utilize a coupled economy-agroecology-hydrology modeling framework to capture the cascading impacts of climate change mitigation policy on agriculture and the resulting water quality cobenefits. We analyze a policy that assigns a range of United States government's social cost of carbon estimates ($51, $76, and $152/ton of CO2-equivalents) to fossil fuel-based CO2 emissions. This policy raises energy costs and, importantly for agriculture, boosts the price of nitrogen fertilizer production. At the highest carbon price, US carbon emissions are reduced by about 50%, and nitrogen fertilizer prices rise by about 90%, leading to an approximate 15% reduction in fertilizer applications for corn production across the Mississippi River Basin. Corn and soybean production declines by about 7%, increasing crop prices by 6%, while nitrate leaching declines by about 10%. Simulated nitrate export to the Gulf of Mexico decreases by 8%, ultimately shrinking the average midsummer area of the Gulf of Mexico hypoxic area by 3% and hypoxic volume by 4%. We also consider the additional benefits of restored wetlands to mitigate nitrogen loading to reduce hypoxia in the Gulf of Mexico and find a targeted wetland restoration scenario approximately doubles the effect of a low to moderate social cost of carbon. Wetland restoration alone exhibited spillover effects that increased nitrate leaching in other parts of the basin which were mitigated with the inclusion of the carbon policy. We conclude that a national climate policy aimed at reducing greenhouse gas emissions in the United States would have important water quality cobenefits.

Investing in nature can improve equity and economic returns.

Sustainable development requires jointly achieving economic development to raise standards of living and environmental sustainability to secure these gains for the long run. Here, we develop a local-to-global, and global-to-local, earth-economy model that integrates the Global Trade Analysis Project (GTAP)-computable general equilibrium model of the economy with the Integrated Valuation of Ecosystem Services and Tradeoffs (InVEST) model of fine-scale, spatially explicit ecosystem services. The integrated model, GTAP-InVEST, jointly determines land use, environmental conditions, ecosystem services, market prices, supply and demand across economic sectors, trade across regions, and aggregate performance metrics like GDP. We use the integrated model to analyze the contribution of investing in nature for economic prosperity, accounting for the impact of four important ecosystem services (pollination, timber provision, marine fisheries, and carbon sequestration). We show that investments in nature result in large improvements relative to a business-as-usual path, accruing annual gains of $100 to $350 billion (2014 USD) with the largest percentage gains in the lowest-income countries. Our estimates include only a small subset of ecosystem services and could be far higher with inclusion of more ecosystem services, incorporation of ecological tipping points, and reduction in substitutability that limits economic adjustments to declines in natural capital. Our analysis highlights the need for improved environmental-economic modeling and the vital importance of integrating environmental information firmly into economic analysis and policy. The benefits of doing so are potentially very large, with the greatest percentage benefits accruing to inhabitants of the poorest countries.

Recent Developments and Challenges in Projecting the Impact of Crop Productivity Growth on Biodiversity Considering Market-Mediated Effects.

The effect of an increase in crop productivity (output per unit of inputs) on biodiversity is hitherto poorly understood. This is because increased productivity of a crop in particular regions leads to increased profit that can encourage expansion of its cultivated area causing land use change and ultimately biodiversity loss, a phenomenon also known as "Jevons paradox" or the "rebound effect". Modeling such consequences in an interconnected and globalized world considering such rebound effects is challenging. Here, we discuss the use of computable general equilibrium (CGE) and other economic models in combination with ecological models to project consequences of crop productivity improvements for biodiversity globally. While these economic models have the advantage of taking into account market-mediated responses, resource constraints, endogenous price responses, and dynamic bilateral patterns of trade, there remain a number of important research and data gaps in these models which must be addressed to improve their performance in assessment of the link between local crop productivity changes and global biodiversity. To this end, we call for breaking the silos and building interdisciplinary networks across the globe to facilitate data sharing and knowledge exchange in order to improve global-to-local-to-global analysis of land, biodiversity, and ecosystem sustainability.

Market-mediated responses confound policies to limit deforestation from oil palm expansion in Malaysia and Indonesia.

The global demand for palm oil has grown rapidly over the past several decades. Much of the output expansion has occurred in carbon- and biodiversity-rich forest lands of Malaysia and Indonesia (M&I), contributing to record levels of terrestrial carbon emissions and biodiversity loss. This has led to a variety of voluntary and mandatory regulatory actions, as well as calls for limits on palm oil imports from M&I. This paper offers a comprehensive, global assessment of the economic and environmental consequences of alternative policies aimed at limiting deforestation from oil palm expansion in M&I. It highlights the challenges of limiting forest and biodiversity loss in the presence of market-mediated spillovers into related oilseed and agricultural commodity and factor markets, both in M&I and overseas. Indeed, limiting palm oil production or consumption is unlikely to halt deforestation in M&I in the absence of active forest conservation incentives. Policies aimed at restricting palm oil production in M&I also have broader consequences for the economy, including significant impacts on consumer prices, real wages, and welfare, that vary among different global regions. A crucial distinction is whether the initiative is undertaken domestically, in which case the M&I region could benefit, or by major palm oil importers, in which case M&I loses income. Nonetheless, all policies considered here pass the social welfare test of global carbon dioxide mitigation benefits exceeding their costs.

Fine-Scale Analysis of the Energy-Land-Water Nexus: Nitrate Leaching Implications of Biomass Cofiring in the Midwestern United States.

As scientists seek to better understand the linkages between energy, water, and land systems, they confront a critical question of scale for their analysis. Many studies exploring this nexus restrict themselves to a small area in order to capture fine-scale processes, whereas other studies focus on interactions between energy, water, and land over broader domains but apply coarse resolution methods. Detailed studies of a narrow domain can be misleading if the policy intervention considered is broad-based and has impacts on energy, land, and agricultural markets. Regional studies with aggregate low-resolution representations may miss critical feedbacks driven by the dynamic interactions between subsystems. This study applies a novel, gridded energy-land-water modeling system to analyze the local environmental impacts of biomass cofiring of coal power plants across the upper MISO region. We use this framework to examine the impacts of a hypothetical biomass cofiring technology mandate of coal-fired power plants using corn residues. We find that this scenario has a significant impact on land allocation, fertilizer applications, and nitrogen leaching. The effects also impact regions not involved in cofiring through agricultural markets. Further, some MISO coal-fired plants would cease generation because the competition for biomass increases the cost of this feedstock and because the higher operating costs of cofiring renders them uncompetitive with other generation sources. These factors are not captured by analyses undertaken at the level of an individual power plant. We also show that a region-wide analysis of this cofiring mandate would have registered only a modest increase in nitrate leaching (just +5% across the upper MISO region). Such aggregate analyses would have obscured the extremely large increases in leaching at particular locations, as much as +60%. Many of these locations are already pollution hotspots. Fine-scale analysis, nested within a broader framework, is necessary to capture these critical environmental interactions within the energy, land, and water nexus.

Global market integration increases likelihood that a future African Green Revolution could increase crop land use and CO2 emissions.

There has been a resurgence of interest in the impacts of agricultural productivity on land use and the environment. At the center of this debate is the assertion that agricultural innovation is land sparing. However, numerous case studies and global empirical studies have found little evidence of higher yields being accompanied by reduced area. We find that these studies overlook two crucial factors: estimation of a true counterfactual scenario and a tendency to adopt a regional, rather than a global, perspective. This paper introduces a general framework for analyzing the impacts of regional and global innovation on long run crop output, prices, land rents, land use, and associated CO2 emissions. In so doing, it facilitates a reconciliation of the apparently conflicting views of the impacts of agricultural productivity growth on global land use and environmental quality. Our historical analysis demonstrates that the Green Revolution in Asia, Latin America, and the Middle East was unambiguously land and emissions sparing, compared with a counterfactual world without these innovations. In contrast, we find that the environmental impacts of a prospective African Green Revolution are potentially ambiguous. We trace these divergent outcomes to relative differences between the innovating region and the rest of the world in yields, emissions efficiencies, cropland supply response, and intensification potential. Globalization of agriculture raises the potential for adverse environmental consequences. However, if sustained for several decades, an African Green Revolution will eventually become land sparing.

Segmental aortic stiffening contributes to experimental abdominal aortic aneurysm development.

BACKGROUND: Stiffening of the aortic wall is a phenomenon consistently observed in age and in abdominal aortic aneurysm (AAA). However, its role in AAA pathophysiology is largely undefined. METHODS AND RESULTS: Using an established murine elastase-induced AAA model, we demonstrate that segmental aortic stiffening precedes aneurysm growth. Finite-element analysis reveals that early stiffening of the aneurysm-prone aortic segment leads to axial (longitudinal) wall stress generated by cyclic (systolic) tethering of adjacent, more compliant wall segments. Interventional stiffening of AAA-adjacent aortic segments (via external application of surgical adhesive) significantly reduces aneurysm growth. These changes correlate with the reduced segmental stiffness of the AAA-prone aorta (attributable to equalized stiffness in adjacent segments), reduced axial wall stress, decreased production of reactive oxygen species, attenuated elastin breakdown, and decreased expression of inflammatory cytokines and macrophage infiltration, and attenuated apoptosis within the aortic wall, as well. Cyclic pressurization of segmentally stiffened aortic segments ex vivo increases the expression of genes related to inflammation and extracellular matrix remodeling. Finally, human ultrasound studies reveal that aging, a significant AAA risk factor, is accompanied by segmental infrarenal aortic stiffening. CONCLUSIONS: The present study introduces the novel concept of segmental aortic stiffening as an early pathomechanism generating aortic wall stress and triggering aneurysmal growth, thereby delineating potential underlying molecular mechanisms and therapeutic targets. In addition, monitoring segmental aortic stiffening may aid the identification of patients at risk for AAA.

Global climate policy impacts on livestock, land use, livelihoods, and food security.

Recent research has shed light on the cost-effective contribution that agriculture can make to global greenhouse gas abatement; however, the resulting impacts on agricultural production, producer livelihoods, and food security remain largely unexplored. This paper provides an integrated assessment of the linkages between land-based climate policies, development, and food security, with a particular emphasis on abatement opportunities and impacts in the livestock sector. Targeting Annex I countries and exempting non-Annex I countries from land-based carbon policies on equity or food security grounds may result in significant leakage rates for livestock production and agriculture as a whole. We find that such leakage can be eliminated by supplying forest carbon sequestration incentives to non-Annex I countries. Furthermore, substantial additional global agricultural abatement can be attained by extending a greenhouse gas emissions tax to non-Annex I agricultural producers, while compensating them for their additional tax expenses. Because of their relatively large emissions intensities and limited abatement possibilities, ruminant meat producers face the greatest market adjustments to land-based climate policies. We also evaluate the impacts of climate policies on livelihoods and food consumption in developing countries. In the absence of non-Annex I abatement policies, these impacts are modest. However, strong income and food consumption impacts surface because of higher food costs after forest carbon sequestration is promoted at a global scale. Food consumption among unskilled labor households falls but rises for the representative farm households, because global agricultural supplies are restricted and farm prices rise sharply in the face of inelastic food demands.

A fluorescence-based array screen for transglutaminase substrates.

Transglutaminases (EC 2.3.2.13) form an enzyme family that catalyzes the formation of isopeptide bonds between the γ-carboxamide group of glutamine and the ε-amine group of lysine residues of peptides and proteins. Other primary amines can be accepted in place of lysine. Because of their important physiological and pathophysiological functions, transglutaminases have been studied for 60 years. However, the substrate preferences of this enzyme class remain largely elusive. In this study, we used focused combinatorial libraries of 400 peptides to investigate the influence of the amino acids adjacent to the glutamine and lysine residues on the catalysis of isopeptide bond formation by microbial transglutaminase. Using the peptide microarray technology we found a strong positive influence of hydrophobic and basic amino acids, especially arginine, tyrosine, and leucine. Several tripeptide substrates were synthesized, and enzymatic kinetic parameters were determined both by microarray analysis and in solution.

Transglutamination allows production and characterization of native-sized ELPylated spider silk proteins from transgenic plants.

In the last two decades it was shown that plants have a great potential for production of specific heterologous proteins. But high cost and inefficient downstream processing are a main technical bottleneck for the broader use of plant-based production technology especially for protein-based products, for technical use as fibres or biodegradable plastics and also for medical applications. High-performance fibres from recombinant spider silks are, therefore, a prominent example. Spiders developed rather different silk materials that are based on proteins. These spider silks show excellent properties in terms of elasticity and toughness. Natural spider silk proteins have a very high molecular weight, and it is precisely this property which is thought to give them their strength. Transgenic plants were generated to produce ELPylated recombinant spider silk derivatives. These fusion proteins were purified by Inverse Transition Cycling (ITC) and enzymatically multimerized with transglutaminase in vitro. Layers produced by casting monomers and multimers were characterized using atomic force microscopy (AFM) and AFM-based nanoindentation. The layered multimers formed by mixing lysine- and glutamine-tagged monomers were associated with the highest elastic penetration modulus.

Partial remission definition: validation based on the insulin dose-adjusted HbA1c (IDAA1C) in 129 Danish children with new-onset type 1 diabetes.

OBJECTIVE: To validate the partial remission (PR) definition based on insulin dose-adjusted HbA1c (IDAA1c). SUBJECTS AND METHODS: The IDAA1c was developed using data in 251 children from the European Hvidoere cohort. For validation, 129 children from a Danish cohort were followed from the onset of type 1 diabetes (T1D). Receiver operating characteristic curve (ROC) analysis was used to evaluate the predictive value of IDAA1c and age on partial C-peptide remission (stimulated C-peptide, SCP > 300 pmol/L). RESULTS: PR (IDAA1c ≤ 9) in the Danish and Hvidoere cohorts occurred in 62 vs. 61% (3 months, p = 0.80), 47 vs. 44% (6 months, p = 0.57), 26 vs. 32% (9 months, p = 0.32) and 19 vs. 18% (12 months, p = 0.69). The effect of age on SCP was significantly higher in the Danish cohort compared with the Hvidoere cohort (p < 0.0001), likely due to higher attained Boost SCP, so the sensitivity and specificity of those in PR by IDAA1c ≤ 9, SCP > 300 pmol/L was 0.85 and 0.62 at 6 months and 0.62 vs. 0.38 at 12 months, respectively. IDAA1c with age significantly improved the ROC analyses and the AUC reached 0.89 ± 0.04 (age) vs. 0.94 ± 0.02 (age + IDAA1c) at 6 months (p < 0.0004) and 0.76 ± 0.04 (age) vs. 0.90 ± 0.03 (age + IDAA1c) at 12 months (p < 0.0001). CONCLUSIONS: The diagnostic and prognostic power of the IDAA1c measure is kept but due to the higher Boost stimulation in the Danish cohort, the specificity of the formula is lower with the chosen limits for SCP (300 pmol/L) and IDAA1c ≤9, respectively.

Planned expansion of transportation infrastructure in Brazil has implications for the pattern of agricultural production and carbon emissions.

High transportation costs have been a barrier to the expansion of agriculture in the interior of Brazil. To reduce transportation costs, Brazil launched the National Logistics Plan, aiming to expand its railway network by up to 91 % by 2035. Such a large-scale infrastructure investment raises concerns about its economic and environmental consequences. By combining geospatial estimation of transportation cost with a grid-resolving, multi-scale economic model that bridges fine-scale crop production with its trade and demand from national and global perspectives, we explore impacts of transportation infrastructure expansion on agricultural production, land use changes, and carbon emissions both locally and nationally in Brazil. We find that globally, the impacts on output and land use changes are small. However, within Brazil, the plan's primary impacts are impressive. PNL2035 results in the reduction of transportation costs by 8-23 % across states (depending on expansion's extent) in the interior Cerrado biome. This results in cropland expansion and increases in terrestrial carbon emissions in the Cerrado region. However, the increase in terrestrial carbon emissions in the Cerrado is offset by spillover effects elsewhere in Brazil, as crop production shifts away from the Southeast-South regions and accompanying change in the mix of transportation mode for farm products from roadway to more emission-efficient railway. Furthermore, we argue that the transportation infrastructure's impact on the enhanced mobility of labor and other agricultural inputs would further accentuate the regional shift in agricultural production and contribute to carbon emission mitigation. Upon its completion, PNL2035 is expected to result in the reduction of net national emissions by 1.8-30.7 million metric ton of CO2-equivalent, depending on the impacts on labor and purchased input mobility. We conclude that the omission of spillover effects due to infrastructure expansion can lead to misleading assessments of transport policies.

Solutions to the double burden of malnutrition also generate health and environmental benefits.

Present food consumption patterns will intensify pressure on natural resources, while poor nutrition is expected to prevail at both low and high levels of calorie consumption. To better understand the interplay between food security, environment and health, we use an integrated framework that allows for the analysis of the dynamics of the double burden of malnutrition and its health and environmental impacts by 2050. We find that excessive caloric intake will be key in rising body mass index levels, particularly in emerging economies. Because higher levels of body mass index will be reached at younger ages, future cohorts will increase their exposure to health risks, including coronary heart disease, stroke, site-specific cancers and type 2 diabetes. This framework also offers insights into the health, food and environmental security impacts of changing food demand behaviour. We find that reductions in food purchasing-associated with the mitigation of food waste and excessive food intake-are more important than changes in dietary composition in increasing food affordability and reducing pressure on cropland expansion, whereas dietary composition is critical in driving greenhouse gas emissions.

The Arabidopsis thaliana phosphate starvation responsive gene AtPPsPase1 encodes a novel type of inorganic pyrophosphatase.

BACKGROUND: Low inorganic phosphate (Pi) availability triggers metabolic responses to maintain the intracellular phosphate homeostasis in plants. One crucial adaptive mechanism is the immediate cleavage of Pi from phosphorylated substrates; however, phosphohydrolases that function in the cytosol and putative substrates have not been characterized yet. One candidate gene is Arabidopsis thaliana At1g73010 encoding an uncharacterized enzyme with homology to the haloacid dehalogenase (HAD) superfamily. METHODS AND RESULTS: This work reports the molecular cloning of At1g73010, its expression in Escherichia coli, and the enzymatic characterisation of the recombinant protein (33.5 kD). The Mg²(+)-dependent enzyme named AtPPsPase1 catalyzes the specific cleavage of pyrophosphate (K(m) 38.8 µM) with an alkaline catalytic pH optimum. Gel filtration revealed a tetrameric structure of the soluble cytoplasmic protein. Modelling of the active site and assay of the recombinant protein variant D19A demonstrated that the enzyme shares the catalytic mechanism of the HAD superfamily including a phosphorylated enzyme intermediate. CONCLUSIONS: The tight control of AtPPsPase1 gene expression underlines its important role in the Pi starvation response and suggests that cleavage of pyrophosphate is an immediate metabolic adaptation reaction. GENERAL SIGNIFICANCE: The novel enzyme, the first pyrophosphatase in the HAD superfamily, differs from classical pyrophosphatases with respect to structure and catalytic mechanism. The enzyme function could be used to discover unknown aspects of pyrophosphate metabolism in general.

Increased thermostability of microbial transglutaminase by combination of several hot spots evolved by random and saturation mutagenesis.

The thermostability of microbial transglutaminase (MTG) of Streptomyces mobaraensis was further improved by saturation mutagenesis and DNA-shuffling. High-throughput screening was used to identify clones with increased thermostability at 55°C. Saturation mutagenesis was performed at seven "hot spots", previously evolved by random mutagenesis. Mutations at four positions (2, 23, 269, and 294) led to higher thermostability. The variants with single amino acid exchanges comprising the highest thermostabilities were combined by DNA-shuffling. A library of 1,500 clones was screened and variants showing the highest ratio of activities after incubation for 30 min at 55°C relative to a control at 37°C were selected. 116 mutants of this library showed an increased thermostability and 2 clones per deep well plate were sequenced (35 clones). 13 clones showed only the desired sites without additional point mutations and eight variants were purified and characterized. The most thermostable mutant (triple mutant S23V-Y24N-K294L) exhibited a 12-fold higher half-life at 60°C and a 10-fold higher half-life at 50°C compared to the unmodified recombinant wild-type enzyme. From the characterization of different triple mutants differing only in one amino acid residue, it can be concluded that position 294 is especially important for thermostabilization. The simultaneous exchange of amino acids at sites 23, 24, 269 and 289 resulted in a MTG-variant with nearly twofold higher specific activity and a temperature optimum of 55°C. A triple mutant with amino acid substitutions at sites 2, 289 and 294 exhibits a temperature optimum of 60°C, which is 10°C higher than that of the wild-type enzyme.

Investigations on the activation of recombinant microbial pro-transglutaminase: in contrast to proteinase K, dispase removes the histidine-tag.

In order to produce recombinant microbial transglutaminase (rMTG) which is free of the activating protease, dispase was used to activate the pro-rMTG followed by immobilized metal affinity chromatography (IMAC). As shown by MALDI-MS, the dispase does not only cleave the pro-sequence, but unfortunately also cleaves within the C-terminal histidine-tag. Hence, the active rMTG cannot properly bind to the IMAC material. As an alternative, proteinase K was investigated. This protease was successfully applied for the activation of purified pro-rMTG either as free or immobilized enzyme and the free enzyme was also applicable directly in the crude cell extract of E. coli. Thus, it enables a simple two-step activation/purification procedure resulting in protease-free and almost pure transglutaminase preparations. The protocol has been successfully applied to both, wild-type transglutaminase of Streptomyces mobaraensis as well as to the highly active variant S2P. Proteinase K activates the pro-rMTG without unwanted degradation of the histidine-tag. It turned out to be very important to inhibit proteinase K activity, e.g., by PMSF, prior to protein separation by SDS-PAGE.

Deciphering the Biodiversity-Production Mutualism in the Global Food Security Debate.

Without changes in consumption, along with sharp reductions in food waste and postharvest losses, agricultural production must grow to meet future food demands. The variety of concepts and policies relating to yield increases fail to integrate an important constituent of production and human nutrition - biodiversity. We develop an analytical framework to unpack this biodiversity-production mutualism (BPM), which bridges the research fields of ecology and agroeconomics and makes the trade-off between food security and protection of biodiversity explicit. By applying the framework, the incorporation of agroecological principles in global food systems are quantifiable, informed assessments of green total factor productivity (TFP) are supported, and possible lock-ins of the global food system through overintensification and associated biodiversity loss can be avoided.

Cloning, expression, purification, and characterization of a designer protein with repetitive sequences.

An artificial protein containing alternating hydrophilic-hydrophobic blocks of amino acids was designed in order to mimic the structure of synthetic multiblock copolymers. The hydrophobic block consisted of the six amino acids Ala Ile Leu Leu Ile Ile (AILLII) and the hydrophilic block of the eight amino acids Thr Ser Glu Asp Asp Asn Asn Gln (TSEDDNNQ). The coding DNA sequence of the cluster was inserted into an commercial pET 30a(+) vector using a two step strategy. The expression of the artificial protein in Escherichia coli was optimized using a temperature shift strategy. Only at cultivation temperature of 24 degrees C after induction expression was observed, whereas at 30 and 37 degrees C no target protein could be detected. Cells obtained from a 15L bioreactor cultivation of E. coli were disintegrated by mechanical methods. Interestingly, glass bead milling and high pressure homogenization resulted in a different solubility of the target protein. The further purification was carried out by affinity chromatography using the soluble homogenized protein. Extreme conditions (6M urea, 0.5M NaCl) were applied in order to prevent aggregation to insoluble particles. The designer protein showed an extremely high tendency to form dimers or trimers caused by intermolecular interactions which were even not broken under the conditions of SDS-polyacrylamide gel electrophoresis, rendering the behavior during purification different from proteins usually found in nature. The protein preparation was not completely pure according to SDS-PAGE stained by Coomassie blue or silver. In MALDI-TOF-MS, nano-ESI qTOF-MS of the entire protein preparation and nano-ESI-MS after digestion by trypsin and chymotrypsin impurities were not detectable.