Decision Making Methods for Water Resources Management Under Deep Uncertainty

Substantial anthropogenic change of the Earth’s climate is modifying patterns of rainfall, river flow, glacial melt and groundwater recharge rates across the planet, undermining many of the stationarity assumptions upon which water resources infrastructure has been historically managed. This hydrological uncertainty is creating a potentially vast range of possible futures that could threaten the dependability of vital regional water supplies. This, combined with increased urbanisation and rapidly growing regional populations, is putting pressures on finite water resources. One of the greatest international challenges facing decision makers in the water industry is the increasing influences of these “deep” climate change and population growth uncertainties affecting the long-term balance of supply and demand and necessitating the need for adaptive action. Water companies and utilities worldwide are now under pressure to modernise their management frameworks and approaches to decision making in order to identify more sustainable and cost-effective water management adaptations that are reliable in the face of uncertainty. The aim of this thesis is to compare and contrast a range of existing Decision Making Methods (DMMs) for possible application to Water Resources Management (WRM) problems, critically analyse on real-life case studies their suitability for handling uncertainties relating to climate change and population growth and then use the knowledge generated this way to develop a new, resilience-based WRM planning methodology. This involves a critical evaluation of the advantages and disadvantages of a range of methods and metrics developed to improve on current engineering practice, to ultimately compile a list of suitable recommendations for a future framework for WRM adaptation planning under deep uncertainty. This thesis contributes to the growing vital research and literature in this area in several distinct ways. Firstly, it qualitatively reviews a range of DMMs for potential application to WRM adaptation problems using a set of developed criteria. Secondly, it quantitatively assesses two promising and contrasting DMMs on two suitable real-world case studies to compare highlighted aspects derived from the qualitative review and evaluate the adaptation outputs on a practical engineering level. Thirdly, it develops and reviews a range of new potential performance metrics that could be used to quantitatively define system resilience to help answer the water industries question of how best to build in more resilience in future water resource adaptation planning. This leads to the creation and testing of a novel resilience driven methodology for optimal water resource planning, combining optimal aspects derived from the quantitative case study work with the optimal metric derived from the resilience metric investigation. Ultimately, based on the results obtained, a list of suitable recommendations is compiled on how to improve the existing methodologies for future WRM planning under deep uncertainty. These recommendations include the incorporation of more complex simulation models into the planning process, utilisation of multi-objective optimisation algorithms, improved uncertainty characterisation and assessments, an explicit robustness examination and the incorporation of additional performance metrics to increase the clarity of the strategy assessment process.HR Wallingford LtdEngineering and Physical Sciences Research Council (EPSRC

Glycogen and its metabolism: some new developments and old themes

Glycogen is a branched polymer of glucose that acts as a store of energy in times of nutritional sufficiency for utilization in times of need. Its metabolism has been the subject of extensive investigation and much is known about its regulation by hormones such as insulin, glucagon and adrenaline (epinephrine). There has been debate over the relative importance of allosteric compared with covalent control of the key biosynthetic enzyme, glycogen synthase, as well as the relative importance of glucose entry into cells compared with glycogen synthase regulation in determining glycogen accumulation. Significant new developments in eukaryotic glycogen metabolism over the last decade or so include: (i) three-dimensional structures of the biosynthetic enzymes glycogenin and glycogen synthase, with associated implications for mechanism and control; (ii) analyses of several genetically engineered mice with altered glycogen metabolism that shed light on the mechanism of control; (iii) greater appreciation of the spatial aspects of glycogen metabolism, including more focus on the lysosomal degradation of glycogen; and (iv) glycogen phosphorylation and advances in the study of Lafora disease, which is emerging as a glycogen storage disease

Redox Switch for the Inhibited State of Yeast Glycogen Synthase Mimics Regulation by Phosphorylation

Glycogen synthase (GS) is the rate limiting enzyme in the synthesis of glycogen. Eukaryotic GS is negatively regulated by covalent phosphorylation and allosterically activated by glucose-6-phosphate (G6P). To gain structural insights into the inhibited state of the enzyme, we solved the crystal structure of yGsy2-R589A/R592A to a resolution of 3.3 Å. The double mutant has an activity ratio similar to the phosphorylated enzyme and also retains the ability to be activated by G6P. When compared to the 2.88 Å structure of the wild-type G-6-P activated enzyme, the crystal structure of the low-activity mutant showed that the N-terminal domain of the inhibited state is tightly held against the dimer-related interface thereby hindering acceptor access to the catalytic cleft. Based on these two structural observations, we developed a reversible redox regulatory feature in yeast GS by substituting cysteine residues for two highly conserved arginine residues. When oxidized, the cysteine mutant enzyme exhibits activity levels similar to the phosphorylated enzyme, but cannot be activated by G-6-P. Upon reduction, the cysteine mutant enzyme regains normal activity levels and regulatory response to G-6-P activation

Evaluation Of Decision Making Methods For Integrated Water Resource Management Under Uncertainty

Water companies and utilities in the UK are required to produce Water Resource Management Plans (WRMPs) every five years that outline their future strategies for maintaining a secure water supply to meet anticipated demand levels. Regulatory frameworks differ around the world but in most countries similar plans are developed under the auspices of Integrated Water Resources Management (IWRM) programmes. The plans justify new demand management and water supply infrastructure needed and validate management decisions. One of the greatest problems now facing decision makers in the water industry are the increasing uncertainties in the variables used in estimating the supply and demand balance due to increased levels of climate change. WRMPs in the future will need to deliver plans that can adapt water systems to face a widening variation of possible future states; with increased consideration to uncertain water availability, resource deterioration and demand levels. This paper reviews several established decision making methods and analyses their performance and suitability within an IWRM problem. The methods include Info-Gap decision theory, Robust Optimisation, Minimax Regret, Laplace theory and Maximin theory. These methods have been designed to aid decision making under severe uncertainty but differences exist in their approach and attitude to risk. For example, the Info-Gap methodology offers solutions that provide robustness of sufficing over a wide range of uncertainty, but is highly dependent on initial parameter settings. Robust Optimisation concentrates on optimising for robustness over a set of objective functions instead of satisfying a set of constraints. Laplace, Maximin and Minimax Regret are all classical decision methods that implicitly reflect a particular attitude to risk when dealing with severe uncertainty. These methods were applied to a case study resembling the Sussex North region in England, assessing their applicability at improving the IWRM problem and highlighting the strengths and weaknesses of each method

Structural insights into the inhibited state of Glycogen Synthase

poster abstractGlycogen is an osmotically inert polymer of glucose, synthesized during times of nutritional sufficiency so that it can be rapidly catabolized when there is an energy demand1. Glycogen synthase (GS) is responsible for the bulk of its synthesis by transferring glucose from UDPG to an existing glucose polymer1. Eukaryotic GS is allosterically activated by glucose-6-phosphate (G6P) and negatively regulated by covalent phosphorylation2. A cluster of six arginine residues are conserved across all eukaryotic species which determine the enzyme’s ability to respond to these activating and inhibitory signals2. Prior structural studies from our lab had shed light on the dephosphorylated and activated state of the enzyme3. However, little is known on the phosphorylated state of the enzyme. For structural studies on the inhibited state, we used the yGsy2R589/592A mutant as a surrogate since it has a basal activity state similar to the inhibited phosphorylated state. We solved the structure of the mutant to a resolution of 3.3 Å. While the overall structural arrangement of the tetramer is similar to the basal state enzyme, the interfaces are more closed. In particular, the N-terminal Rossmann-fold domain is rotated toward the interface by 5.9°, limiting access to the active site by the acceptor end of the glycogen chain. Coincident with this domain closure, we also observed that the the distance between the regulatory helices of adjacent monomers are moved closer to one another. Based on this observation, we hypothesized we could develop a reversible redox regulatory feature in the enzyme by substituting cysteine residues for arginines 581 and 592, which lie across from each other at the interface. Consistent with our hypothesis, the yGsy2R581/592C double mutant exhibited very low activity, and could not be activated by G6P. However, normal function of the enzyme could be restored in the presence of reducing agents like DTT, BME and TCEP. Taken together, our mutational work demonstrates that the conserved arginine cluster in the regulatory helix, both regulates the enzyme’s response to signaling inputs and keeps the enzyme in a basal state conformation that is poised to respond to the activating and inhibitory inputs

Incorporation of phosphate into glycogen by glycogen synthase

The storage polymer glycogen normally contains small amounts of covalently attached phosphate as phosphomonoesters at C2, C3 and C6 atoms of glucose residues. In the absence of the laforin phosphatase, as in the rare childhood epilepsy Lafora disease, the phosphorylation level is elevated and is associated with abnormal glycogen structure that contributes to the pathology. Laforin therefore likely functions in vivo as a glycogen phosphatase. The mechanism of glycogen phosphorylation is less well-understood. We have reported that glycogen synthase incorporates phosphate into glycogen via a rare side reaction in which glucose-phosphate rather than glucose is transferred to a growing polyglucose chain (Tagliabracci et al. (2011) Cell Metab13, 274-282). We proposed a mechanism to account for phosphorylation at C2 and possibly at C3. Our results have since been challenged (Nitschke et al. (2013) Cell Metab17, 756-767). Here we extend the evidence supporting our conclusion, validating the assay used for the detection of glycogen phosphorylation, measurement of the transfer of (32)P from [β-(32)P]UDP-glucose to glycogen by glycogen synthase. The (32)P associated with the glycogen fraction was stable to ethanol precipitation, SDS-PAGE and gel filtration on Sephadex G50. The (32)P-signal was not affected by inclusion of excess unlabeled UDP before analysis or by treatment with a UDPase, arguing against the signal being due to contaminating [β-(32)P]UDP generated in the reaction. Furthermore, [(32)P]UDP did not bind non-covalently to glycogen. The (32)P associated with glycogen was released by laforin treatment, suggesting that it was present as a phosphomonoester. The conclusion is that glycogen synthase can mediate the introduction of phosphate into glycogen, thereby providing a possible mechanism for C2, and perhaps C3, phosphorylation

A Non-Anesthetized Mouse Model for Recording Sensory Urinary Bladder Activity

The goal of this study was to develop an in vivo awake mouse model for extracellular bladder sensory nerve recording. A bipolar 125-μm silver electrode was positioned under a single postganglionic bladder nerve. Efferent nerve signals were eliminated by tying off the postganglionic bladder nerve between the major pelvic ganglion and the recording electrode. Sensory nerve activity was measured in the conscious animals 48 h after surgery during continuous intravesical infusion of 0.9% saline/0.5% acetic acid followed by 0.5% acetic acid with capsazepine (10 μM) at a rate of 0.75 ml/h. Continuous infusion of 0.9% NaCl led to a gradual increase in the frequency of sensory nerve firing that peaked upon reaching threshold pressure. Non-micturition contractions were observed in some animals during filling and other animals exhibited only minimal pressure fluctuations; both types of events were associated with a rise in sensory nerve activity. Intravesical infusion of 0.5% acetic acid reduced the intermicturition interval. This was associated with a 2.1-fold increase in bladder pressure during filling and a two-fold increase at both threshold and micturition pressures. Concurrent with these changes, sensory activity increased 2.8-fold during filling and 2.4-fold at threshold pressure. Subsequent intravesical infusion of capsazepine in 0.5% acetic acid reduced filling and threshold pressures by 21 and 31.2%, respectively, and produced corresponding decreases of 36 and 23.4% in sensory nerve activity. The current study shows that multifiber sensory nerve recordings can be reproducibly obtained from conscious mice

Projecting coral responses to intensifying marine heatwaves under ocean acidification

Over this century, coral reefs will run the gauntlet of climate change, as marine heatwaves (MHWs) become more intense and frequent, and ocean acidification (OA) progresses. However, we still lack a quantitative assessment of how, and to what degree, OA will moderate the responses of corals to MHWs as they intensify throughout this century. Here, we first projected future MHW intensities for tropical regions under three future greenhouse gas emissions scenario (representative concentration pathways, RCP2.6, RCP4.5 and RCP8.5) for the near-term (2021–2040), mid-century (2041–2060) and late-century (2081–2100). We then combined these MHW intensity projections with a global data set of 1,788 experiments to assess coral attribute performance and survival under the three emissions scenarios for the near-term, mid-century and late-century in the presence and absence of OA. Although warming and OA had predominately additive impacts on the coral responses, the contribution of OA in affecting most coral attributes was minor relative to the dominant role of intensifying MHWs. However, the addition of OA led to greater decreases in photosynthesis and survival under intermediate and unrestricted emissions scenario for the mid- and late-century than if intensifying MHWs were considered as the only driver. These results show that role of OA in modulating coral responses to intensifying MHWs depended on the focal coral attribute and extremity of the scenario examined. Specifically, intensifying MHWs and OA will cause increasing instances of coral bleaching and substantial declines in coral productivity, calcification and survival within the next two decades under the low and intermediate emissions scenario. These projections suggest that corals must rapidly adapt or acclimatize to projected ocean conditions to persist, which is far more likely under a low emissions scenario and with increasing efforts to manage reefs to enhance resilience

Integrating environmental variability to broaden the research on coral responses to future ocean conditions

Our understanding of the response of reef-building corals to changes in their physical environment is largely based on laboratory experiments, analysis of long-term field data, and model projections. Experimental data provide unique insights into how organisms respond to variation of environmental drivers. However, an assessment of how well experimental conditions cover the breadth of environmental conditions and variability where corals live successfully is missing. Here, we compiled and analyzed a globally distributed dataset of in-situ seasonal and diurnal variability of key environmental drivers (temperature, pCO2, and O2) critical for the growth and livelihood of reef-building corals. Using a meta-analysis approach, we compared the variability of environmental conditions assayed in coral experimental studies to current and projected conditions in their natural habitats. We found that annual temperature profiles projected for the end of the 21st century were characterized by distributional shifts in temperatures with warmer winters and longer warm periods in the summer, not just peak temperatures. Furthermore, short-term hourly fluctuations of temperature and pCO2 may regularly expose corals to conditions beyond the projected average increases for the end of the 21st century. Coral reef sites varied in the degree of coupling between temperature, pCO2, and dissolved O2, which warrants site-specific, differentiated experimental approaches depending on the local hydrography and influence of biological processes on the carbonate system and O2 availability. Our analysis highlights that a large portion of the natural environmental variability at short and long timescales is underexplored in experimental designs, which may provide a path to extend our understanding on the response of corals to global climate change

Discovery and Development of Small-Molecule Inhibitors of Glycogen Synthase

The overaccumulation of glycogen appears as a hallmark in various glycogen storage diseases (GSDs), including Pompe, Cori, Andersen, and Lafora disease. Accumulating evidence suggests that suppression of glycogen accumulation represents a potential therapeutic approach for treating these GSDs. Using a fluorescence polarization assay designed to screen for inhibitors of the key glycogen synthetic enzyme, glycogen synthase (GS), we identified a substituted imidazole, (rac)-2-methoxy-4-(1-(2-(1-methylpyrrolidin-2-yl)ethyl)-4-phenyl-1H-imidazol-5-yl)phenol (H23), as a first-in-class inhibitor for yeast GS 2 (yGsy2p). Data from X-ray crystallography at 2.85 Å, as well as kinetic data, revealed that H23 bound within the uridine diphosphate glucose binding pocket of yGsy2p. The high conservation of residues between human and yeast GS in direct contact with H23 informed the development of around 500 H23 analogs. These analogs produced a structure–activity relationship profile that led to the identification of a substituted pyrazole, 4-(4-(4-hydroxyphenyl)-3-(trifluoromethyl)-1H-pyrazol-5-yl)pyrogallol, with a 300-fold improved potency against human GS. These substituted pyrazoles possess a promising scaffold for drug development efforts targeting GS activity in GSDs associated with excess glycogen accumulation