A metabolism perspective on alternative urban water servicing options using water mass balance

Urban areas will need to pursue new water servicing options to ensure local supply security. Decisions about how best to employ them are not straightforward due to multiple considerations and the potential for problem shifting among them. We hypothesise that urban water metabolism evaluation based a water mass balance can help address this, and explore the utility of this perspective and the new insights it provides about water servicing options. Using a water mass balance evaluation framework, which considers direct urban water flows (both ‘natural’ hydrological and ‘anthropogenic’ flows), as well as water-related energy, we evaluated how the use of alternative water sources (stormwater/rainwater harvesting, wastewater/greywater recycling) at different scales influences the ‘local water metabolism’ of a case study urban development. New indicators were devised to represent the water-related ‘resource efficiency’ and ‘hydrological performance’ of the urban area. The new insights gained were the extent to which alternative water supplies influence the water efficiency and hydrological performance of the urban area, and the potential energy trade-offs. The novel contribution is the development of new indicators of urban water resource performance that bring together considerations of both the ‘anthropogenic’ and ‘natural’ water cycles, and the interactions between them. These are used for the first time to test alternative water servicing scenarios, and to provide a new perspective to complement broader sustainability assessments of urban water

An inclusive city water account by integrating multiple data sources for South-East Queensland (SEQ), Australia

Cities are the hotspots of impacts on local and distant water resources through economic activity and consumption. More than half of the world's population lives in cities, which is expected to reach around two-thirds by 2050. Such a high level of increased urbanization calls for higher attention towards inclusive, safe, resilient, and sustainable cities (Sustainable Development Goals 11). To evaluate sustainability, inclusiveness, and resiliency pathways, a variety of sustainability indicators have been proposed, including the water footprint. The water footprint is defined as the total volume of freshwater used for the goods and services consumed. It covers both direct (e.g. drinking and cleaning) and virtual water flows (water used in the goods and services supply chain, hence also known as embedded water). Virtual water flows through products and services produced in other locations using their water resources influence the function, prosperity, and growth of the cities. Yet, this aspect is absent in the sustainability and strategic city water footprint reduction goals of Australian cities. To fully account for the water dependencies of Australian cities, direct and virtual water flows need to be known. To this purpose, we build inclusive city water of South-East Queensland (SEQ) by combining material flow analysis (MFA) and the multiregional input-output (MRIO) model. Water consumption in SEQ is used to quantify the water footprint on local water resources and net blue virtual water import. Together, this constitutes the water footprint on national water resources. Our results show that the water footprint of SEQ on local water resources is 620 GL with a net virtual water import of 1382 GL. Therefore, the water footprint of SEQ on national water resources is 2002 GL. The water footprint of SEQ on local water resources consists of direct water consumption by households (192 GL) and the industrial sector (428 GL). The consumed direct water of the SEQ industrial sector flows as virtual water to SEQ (149 GL), the rest of Australia (RoAUS) (all other regions except SEQ) (211 GL), and the rest of the world (68 GL). The virtual water inflows breakdown by source regions showed that 386 GL, 1019 GL, and 256 GL of virtual water imported from the major cities (Sydney, Melbourne, Adelaide, and Perth); regional areas of NSW, Victoria, and QLD; and RoAUS, respectively. Overall, the proposed inclusive city water account can enhance subnational estimates of city water footprint for benchmarking, as well as inclusive and resilient city water planning

BCL-2 Expression is Prognostic for Improved Survival in Non-small Cell Lung Cancer

ObjectiveWe used a large patient population to identify immunohistochemical biomarkers to enable improved prognostication in patients with non-small cell lung carcinoma (NSCLC).MethodsA tissue microarray was constructed using duplicate 0.6 mm cores of formalin-fixed paraffin-embedded tissue blocks from 609 patients with NSCLC. Immunohistochemical was used to detect 11 biomarkers including epidermal growth factor receptor, Her2, Her3, p53, p63, bcl-1, bcl-2, Thyroid transcription factor, carcinoembryonic antigen, chromogranin, and synaptophysin. A clinical database was generated prospectively at the time of tissue collection. Survival outcomes were obtained from a Provincial Cancer Registry database. Univariate and multivariate analyses were performed to look for a relationship between biomarker expression, smoking history, and survival.ResultsSurvival data for 535 cases were available. As of June 2005, 429 patients (80%) had died; of these 286 (54%) died of lung cancer, 117 (22%) died of other known causes, and for 26 (5%) the cause of death was not available. Univariate analysis revealed that bcl-2 (p = 0.007) was the only biomarker prognostic for improved overall survival (OS). bcl-2 (p = 0.021) and p63 (p = 0.025) were both found to be prognostic for improved disease-specific survival (DSS). Multivariate analysis (using age and biomarker expression) revealed that bcl-2 expression is prognostic for improved OS (p = 0.005) and DSS (p = 0.021).ConclusionsOur results suggest that bcl-2 expression is prognostic for improved OS and DSS in NSCLC. Testing for bcl-2 expression in a prospective study will help to determine its clinical relevance in prognostication

Innovation and deadlock in governing disasters and climate change collaboratively - Lessons from the Northern Rivers region of New South Wales, Australia

Both scholars and global policy point to the need for, and effectiveness of, locally-led and collaborative disaster and climate change practices. However, there is a need for analysing how these collaborative approaches are developed and used in practice in different contexts. This paper outlines the use of collaborative approaches by a climate change and disaster community of practice across seven local government areas in the Northern Rivers region, New South Wales, Australia. The region has experienced multiple large-scale flood and bushfire disasters since 2017. This ethnographic study uses established collaborative governance and adaptive governance theoretical frameworks to draw findings from: lived experiences of researchers, 22 interviews with diverse stakeholder groups, in-person and online events and the first author's research diary. The findings indicate that collaborative practice in response to disasters and climate change has resulted in both effective outcomes and multi-dimensional challenges, which have lasting implications for communities across the Northern Rivers region. Three critical leverage points for enhanced collaboration and effectiveness were identified. First, improved use and flow of information such as climate projections and place- and needs-based information. Second, mindset shifts that value community knowledge and contributions, and improved practice through enhancing skills in community development and community-led recovery in disaster and climate change practitioners. Lastly, collectively moving from reactive to proactive responses to climate change and disasters. These insights provide an opportunity for improving the design of community-based risk reduction programs and multi-stakeholder governance arrangements into the future, in the state of New South Wales and beyond

Yeasts and wine off-flavours: a technological perspective

Review article. Part of the special issue "Wine microbiology and safety: from the vineyard to the bottle (Microsafety Wine)", 19-20 Nov. 2009, ItalyIn wine production, yeasts have both beneficial and detrimental activities. Saccharomyces cerevisiae is the yeast mainly responsible for turning grape juice into wine but this species and several others may also show undesirable effects in wines. Among such effects, technologists are particularly concerned with the production of offflavours that may occur during all stages of winemaking. Typical spoiling activities include the production of ethyl acetate by apiculate yeasts before fermentation, hydrogen sulphide by S. cerevisiae during fermentation phases, acetaldehyde by film-forming yeasts during bulk storage, and volatile phenols by Dekkera bruxellensis during storage or after bottling. The occurrence of these hazards depends on the technological operations designed to obtain a given type of wine and most can be avoided by current preventive or curative measures. On the contrary, good manufacturing practices must be strengthened to deal with the problem of volatile phenol production in red wines. Appropriate monitoring of D. bruxellensis populations and quantification of 4-ethylphenol is advised during storage, particularly when oak barrels are used, and absence of viable cells must be guaranteed in bottled wines. This work, which is based on our experience at winery level, aims to provide information on appropriate technological strategies to deal with the problem of off-flavours produced by yeasts

Molecular control of sucrose utilization in Escherichia coli W, an efficient sucrose-utilizing strain

Sucrose is an industrially important carbon source for microbial fermentation. Sucrose utilization in Escherichia coli, however, is poorly understood, and most industrial strains cannot utilize sucrose. The roles of the chromosomally encoded sucrose catabolism (csc) genes in E. coli W were examined by knockout and overexpression experiments. At low sucrose concentrations, the csc genes are repressed and cells cannot grow. Removal of either the repressor protein (cscR) or the fructokinase (cscK) gene facilitated derepression. Furthermore, combinatorial knockout of cscR and cscK conferred an improved growth rate on low sucrose. The invertase (cscA) and sucrose transporter (cscB) genes are essential for sucrose catabolism in E. coli W, demonstrating that no other genes can provide sucrose transport or inversion activities. However, cscK is not essential for sucrose utilization. Fructose is excreted into the medium by the cscK-knockout strain in the presence of high sucrose, whereas at low sucrose (when carbon availability is limiting), fructose is utilized by the cell. Overexpression of cscA, cscAK, or cscAB could complement the W Delta cscRKAB knockout mutant or confer growth on a K-12 strain which could not naturally utilize sucrose. However, phenotypic stability and relatively good growth rates were observed in the K-12 strain only when overexpressing cscAB, and full growth rate complementation in W Delta cscRKA Balso required cscAB. Our understanding of sucrose utilization can be used to improve E. coli Wand engineer sucrose utilization in strains which do not naturally utilize sucrose, allowing substitution of sucrose for other, less desirable carbon sources in industrial fermentations

Parallel evolution of the make–accumulate–consume strategy in Saccharomyces and Dekkera yeasts

Saccharomyces yeasts degrade sugars to two-carbon components, in particular ethanol, even in the presence of excess oxygen. This characteristic is called the Crabtree effect and is the background for the 'make–accumulate–consume' life strategy, which in natural habitats helps Saccharomyces yeasts to out-compete other microorganisms. A global promoter rewiring in the Saccharomyces cerevisiae lineage, which occurred around 100 mya, was one of the main molecular events providing the background for evolution of this strategy. Here we show that the Dekkera bruxellensis lineage, which separated from the Saccharomyces yeasts more than 200 mya, also efficiently makes, accumulates and consumes ethanol and acetic acid. Analysis of promoter sequences indicates that both lineages independently underwent a massive loss of a specific cis-regulatory element from dozens of genes associated with respiration, and we show that also in D. bruxellensis this promoter rewiring contributes to the observed Crabtree effect

Regulation of pH by Carbonic Anhydrase 9 Mediates Survival of Pancreatic Cancer Cells With Activated KRAS in Response to Hypoxia.

Background & Aims Most pancreatic ductal adenocarcinomas (PDACs) express an activated form of KRAS, become hypoxic and dysplastic, and are refractory to chemo and radiation therapies. To survive in the hypoxic environment, PDAC cells upregulate enzymes and transporters involved in pH regulation, including the extracellular facing carbonic anhydrase 9 (CA9). We evaluated the effect of blocking CA9, in combination with administration of gemcitabine, in mouse models of pancreatic cancer. Methods We knocked down expression of KRAS in human (PK-8 and PK-1) PDAC cells with small hairpin RNAs. Human and mouse (KrasG12D/Pdx1-Cre/Tp53/RosaYFP) PDAC cells were incubated with inhibitors of MEK (trametinib) or extracellular signal-regulated kinase (ERK), and some cells were cultured under hypoxic conditions. We measured levels and stability of the hypoxia-inducible factor 1 subunit alpha (HIF1A), endothelial PAS domain 1 protein (EPAS1, also called HIF2A), CA9, solute carrier family 16 member 4 (SLC16A4, also called MCT4), and SLC2A1 (also called GLUT1) by immunoblot analyses. We analyzed intracellular pH (pHi) and extracellular metabolic flux. We knocked down expression of CA9 in PDAC cells, or inhibited CA9 with SLC-0111, incubated them with gemcitabine, and assessed pHi, metabolic flux, and cytotoxicity under normoxic and hypoxic conditions. Cells were also injected into either immune-compromised or immune-competent mice and growth of xenograft tumors was assessed. Tumor fragments derived from patients with PDAC were surgically ligated to the pancreas of mice and the growth of tumors was assessed. We performed tissue microarray analyses of 205 human PDAC samples to measure levels of CA9 and associated expression of genes that regulate hypoxia with outcomes of patients using the Cancer Genome Atlas database. Results Under hypoxic conditions, PDAC cells had increased levels of HIF1A and HIF2A, upregulated expression of CA9, and activated glycolysis. Knockdown of KRAS in PDAC cells, or incubation with trametinib, reduced the posttranscriptional stabilization of HIF1A and HIF2A, upregulation of CA9, pHi, and glycolysis in response to hypoxia. CA9 was expressed by 66% of PDAC samples analyzed; high expression of genes associated with metabolic adaptation to hypoxia, including CA9, correlated with significantly reduced survival times of patients. Knockdown or pharmacologic inhibition of CA9 in PDAC cells significantly reduced pHi in cells under hypoxic conditions, decreased gemcitabine-induced glycolysis, and increased their sensitivity to gemcitabine. PDAC cells with knockdown of CA9 formed smaller xenograft tumors in mice, and injection of gemcitabine inhibited tumor growth and significantly increased survival times of mice. In mice with xenograft tumors grown from human PDAC cells, oral administration of SLC-0111 and injection of gemcitabine increased intratumor acidosis and increased cell death. These tumors, and tumors grown from PDAC patient-derived tumor fragments, grew more slowly than xenograft tumors in mice given control agents, resulting in longer survival times. In KrasG12D/Pdx1-Cre/Tp53/RosaYFP genetically modified mice, oral administration of SLC-0111 and injection of gemcitabine reduced numbers of B cells in tumors. Conclusions In response to hypoxia, PDAC cells that express activated KRAS increase expression of CA9, via stabilization of HIF1A and HIF2A, to regulate pH and glycolysis. Disruption of this pathway slows growth of PDAC xenograft tumors in mice and might be developed for treatment of pancreatic cancer