An Ecological Function Approach to Managing Harmful Cyanobacteria in Three Oregon Lakes: Beyond Water Quality Advisories and Total Maximum Daily Loads (TMDLs).

The Oregon Department of Environmental Quality (ODEQ) uses Total Maximum Daily Load (TMDL) calculations, and the associated regulatory process, to manage harmful cyanobacterial blooms (CyanoHABs) attributable to non-point source (NPS) pollution. TMDLs are based on response (lagging) indicators (e.g., measurable quantities of NPS (nutrients: nitrogen {N} and phosphorus {P}), and/or sediment), and highlight the negative outcomes (symptoms) of impaired water quality. These response indicators belatedly address water quality issues, if the cause is impaired riparian functions. Riparian functions assist in decreasing the impacts of droughts and floods (through sequestration of nutrients and excess sediment), allow water to remain on the land surface, improve aquatic habitats, improve water quality, and provide a focus for monitoring and adaptive management. To manage water quality, the focus must be on the drivers (leading indicators) of the causative mechanisms, such as loss of ecological functions. Success in NPS pollution control, and maintaining healthy aquatic habitats, often depends on land management/land use approaches, which facilitate the natural recovery of stream and wetland riparian functions. Focusing on the drivers of ecosystem functions (e.g., vegetation, hydrology, soil, and landform), instead of individual mandated response indicators, using the proper functioning condition (PFC) approach, as a best management practice (BMP), in conjunction with other tools and management strategies, can lead to pro-active policies and approaches, which support positive change in an ecosystem or watershed, and in water quality improvement.

Assessing Dungeness River Functionality and Effectiveness of Best Management Practices (BMPs) Using an Ecological Functional Approach.

Effective stream and wetland Best Management Practices (BMPs) restore the physical processes associated with ecological functions to their Proper Functioning Condition (PFC, i.e., the highest attainable ecological status of a riparian area without consideration of economic, administrative, or social constraints). Ecological functions connect stream monitoring and management to mitigate the causes of ecosystem degradation and enhance restoration. The ecological function approach supports sustainable management of many ecosystem services including water quality, water stability (aquifer recharge), and fish and wildlife habitats. The 1993 Forest Ecosystem Management Assessment Team (FEMAT) report listed the Dungeness River as a Tier 1 key watershed, noted that watersheds are the logical spatial unit for ecosystem management, and that watersheds are important in species management, and understanding the interdependence of physical processes. Watersheds are at the spatial scale where physical and biological disturbances can be observed, and where management constraints and planning options for restoration objectives and strategies can be readily assessed. The US Forest Service (USFS) developed a management strategy for the Middle Dungeness River, and in the 1990s, the Upper Dungeness River was listed as impaired due to sediment, which initiated a US Forest Service change to land management practices. The Lower Dungeness River and bay are listed as impaired due to fecal coliform contamination. Assessing and monitoring the drivers of ecosystem function (vegetation, hydrology, soil, and landform) as part of a watershed adaptive management plan, and implementing BMPs to increase ecological functions, will improve aquatic habitat and water quality. Most BMPs, such as Total Maximum Daily Loads (TMDLs), attempt to improve water quality by reducing the amount of external pollutants reaching the impacted waterbodies, but do not focus on improving the watershed functions. The Proper Functioning Condition (PFC) approach is used to examine the condition of wetlands and streams and provide guidance for quantitative approaches (e.g., TMDL, remote sensing) used in watershed restoration. Improving watershed functions is a BMP that facilitates increased flows of water, nutrients, sediment, and other materials, and improves habitat quality. Using improved watershed functions as a BMP, facilitated by the use of remote sensing, TMDLs, and the PFC methodology is a more effective means of reducing risks across a watershed than by using TMDLs alone.

Ecological risk based assessment used to restore riparian physical functions to a fresh water Creek.

The purpose of this study was to determine if an interdisciplinary team using a qualitative proper functioning condition (PFC) assessment protocol could identify and reverse significant detrimental ecological alterations which occurred within Gertie's Creek watershed, Ontario, Canada. At potential, Gertie's Creek supported a woody debris glacial outwash fine gravel substrate fish spawning habitat. The anthropogenic activities on Georgina Island caused a denuded anadromous fish population since the early-to mid-1990's in the Gertie's Creek watershed. The PFC assessment indicated that anthropogenic activities on Georgina Island negatively impacted stream flows in Gertie's Creek. Reduced stream flow resulted in the natural stream (lotic) riparian habitat not advancing out of an early seral silver maple and eastern hemlock vegetated swamp (forested wetland) habitat. The Gertie's Creek interdisciplinary team PFC assessment indicated that the entire watershed is not in balance with the water and sediment being supplied along with a lack of diverse riparian vegetation. Sediment was not being transported to the wetland and lake coastal areas because of chronic reduced flows. Further qualitative assessments by the authors of other smaller lentic and lotic ecosystems on Georgina Island indicate that reduced hydrologic flow is an issue for the entire island. Ecosystem function management planning works with the ecosystem to continually respond as the ecology changes in ways that enhance remarkable natural recovery.

An ecological function and services approach to total maximum daily load (TMDL) prioritization.

Prioritizing total maximum daily load (TMDL) development starts by considering the scope and severity of water pollution and risks to public health and aquatic life. Methodology using quantitative assessments of in-stream water quality is appropriate and effective for point source (PS) dominated discharge, but less so in watersheds with mostly nonpoint source (NPS) related impairments. For NPSs, prioritization in TMDL development and implementation of associated best management practices should focus on restoration of ecosystem physical functions, including how restoration effectiveness depends on design, maintenance and placement within the watershed. To refine the approach to TMDL development, regulators and stakeholders must first ask if the watershed, or ecosystem, is at risk of losing riparian or other ecologically based physical attributes and processes. If so, the next step is an assessment of the spatial arrangement of functionality with a focus on the at-risk areas that could be lost, or could, with some help, regain functions. Evaluating stream and wetland riparian function has advantages over the traditional means of water quality and biological assessments for NPS TMDL development. Understanding how an ecosystem functions enables stakeholders and regulators to determine the severity of problem(s), identify source(s) of impairment, and predict and avoid a decline in water quality. The Upper Reese River, Nevada, provides an example of water quality impairment caused by NPS pollution. In this river basin, stream and wetland riparian proper functioning condition (PFC) protocol, water quality data, and remote sensing imagery were used to identify sediment sources, transport, distribution, and its impact on water quality and aquatic resources. This study found that assessments of ecological function could be used to generate leading (early) indicators of water quality degradation for targeting pollution control measures, while traditional in-stream water quality monitoring lagged in response to the deterioration in ecological functions.

Quantifying structural physical habitat attributes using LIDAR and hyperspectral imagery.

Structural physical habitat attributes include indices of stream size, channel gradient, substrate size, habitat complexity, and riparian vegetation cover and structure. The Environmental Monitoring and Assessment Program (EMAP) is designed to assess the status and trends of ecological resources at different scales. High-resolution remote sensing provides unique capabilities in detecting a variety of features and indicators of environmental health and condition. LIDAR is an airborne scanning laser system that provides data on topography, channel dimensions (width, depth), slope, channel complexity (residual pools, volume, morphometric complexity, hydraulic roughness), riparian vegetation (height and density), dimensions of riparian zone, anthropogenic alterations and disturbances, and channel and riparian interaction. Hyperspectral aerial imagery offers the advantage of high spectral and spatial resolution allowing for the detection and identification of riparian vegetation and natural and anthropogenic features at a resolution not possible with satellite imagery. When combined, or fused, these technologies comprise a powerful geospatial data set for assessing and monitoring lentic and lotic environmental characteristics and condition.

Multi-scale landscape factors influencing stream water quality in the state of Oregon.

Enterococci bacteria are used to indicate the presence of human and/or animal fecal materials in surface water. In addition to human influences on the quality of surface water, a cattle grazing is a widespread and persistent ecological stressor in the Western United States. Cattle may affect surface water quality directly by depositing nutrients and bacteria, and indirectly by damaging stream banks or removing vegetation cover, which may lead to increased sediment loads. This study used the State of Oregon surface water data to determine the likelihood of animal pathogen presence using enterococci and analyzed the spatial distribution and relationship of biotic (enterococci) and abiotic (nitrogen and phosphorous) surface water constituents to landscape metrics and others (e.g. human use, percent riparian cover, natural covers, grazing, etc.). We used a grazing potential index (GPI) based on proximity to water, land ownership and forage availability. Mean and variability of GPI, forage availability, stream density and length, and landscape metrics were related to enterococci and many forms of nitrogen and phosphorous in standard and logistic regression models. The GPI did not have a significant role in the models, but forage related variables had significant contribution. Urban land use within stream reach was the main driving factor when exceeding the threshold (> or =35 cfu/100 ml), agriculture was the driving force in elevating enterococci in sites where enterococci concentration was <35 cfu/100 ml. Landscape metrics related to amount of agriculture, wetlands and urban all contributed to increasing nutrients in surface water but at different scales. The probability of having sites with concentrations of enterococci above the threshold was much lower in areas of natural land cover and much higher in areas with higher urban land use within 60 m of stream. A 1% increase in natural land cover was associated with a 12% decrease in the predicted odds of having a site exceeding the threshold. Opposite to natural land cover, a one unit change in each of manmade barren and urban land use led to an increase of the likelihood of exceeding the threshold by 73%, and 11%, respectively. Change in urban land use had a higher influence on the likelihood of a site exceeding the threshold than that of natural land cover.

Insulin represses phosphoenolpyruvate carboxykinase gene transcription by causing the rapid disruption of an active transcription complex: a potential epigenetic effect.

Insulin represses gluconeogenesis, in part, by inhibiting the transcription of genes that encode rate-determining enzymes, such as phosphoenolpyruvate carboxykinase (PEPCK) and glucose-6-phosphatase (G-6-Pase). Glucocorticoids stimulate expression of the PEPCK gene but the repressive action of insulin is dominant. Here, we show that treatment of H4IIE hepatoma cells with the synthetic glucocorticoid, dexamethasone (dex), induces the accumulation of glucocorticoid receptor, as well as many transcription factors, coregulators, and RNA polymerase II, on the PEPCK gene promoter. The addition of insulin to dex-treated cells causes the rapid dissociation of glucocorticoid receptor, polymerase II, and several key transcriptional regulators from the PEPCK gene promoter. These changes are temporally related to the reduced rate of PEPCK gene transcription. A similar disruption of the G-6-Pase gene transcription complex was observed. Additionally, insulin causes the rapid demethylation of arginine-17 on histone H3 of both genes. This rapid, insulin-induced, histone demethylation is temporally related to the disruption of the PEPCK and G-6-Pase gene transcription complex, and may be causally related to the mechanism by which insulin represses transcription of these genes.

Estrogen-related receptor alpha is a repressor of phosphoenolpyruvate carboxykinase gene transcription.

The orphan nuclear receptor estrogen-related receptor (ERR) alpha is a downstream effector of the transcriptional coactivator PGC-1alpha in the regulation of genes important for mitochondrial oxidative capacity. PGC-1alpha is also a potent activator of the transcriptional program required for hepatic gluconeogenesis, and in particular of the key gluconeogenic enzyme phosphoenolpyruvate carboxykinase (PEPCK). We report here that the regulatory sequences of the PEPCK gene harbor a functional ERRalpha binding site. However, in contrast to the co-stimulating effects of ERRalpha and PGC-1alpha on mitochondrial gene expression, ERRalpha acts as a transcriptional repressor of the PEPCK gene. Suppression of ERRalpha expression by small interfering RNA leads to reduced binding of ERRalpha to the endogenous PEPCK gene, and an increase in promoter occupancy by PGC-1alpha, suggesting that part of the ERRalpha function at this gene is to antagonize the action of PGC-1alpha. In agreement with the in vitro studies, animals that lack ERRalpha show increased expression of gluconeogenic genes, including PEPCK and glycerol kinase, but decreased expression of mitochondrial genes, such as ATP synthase subunit beta and cytochrome c-1. Our findings suggest that ERRalpha has opposing effects on genes important for mitochondrial oxidative capacity and gluconeogenesis. The different functions of ERRalpha in the regulation of these pathways suggest that enhancing ERRalpha activity could have beneficial effects on glucose metabolism in diabetic subjects by two distinct mechanisms: increasing mitochondrial oxidative capacity in peripheral tissues and liver, and suppressing hepatic glucose production.

The synergistic effect of dexamethasone and all-trans-retinoic acid on hepatic phosphoenolpyruvate carboxykinase gene expression involves the coactivator p300.

Activation of phosphoenolpyruvate carboxykinase (PEPCK) gene transcription in response to all-trans-retinoic acid (RA) or a glucocorticoid such as dexamethasone (Dex) requires a distinct arrangement of DNA-response elements and their cognate transcription activators on the gene promoter. Two of the accessory factor-binding elements involved in the Dex response (gAF1 and gAF3) coincide with the DNA-response elements involved in the RA response. We demonstrate here that the combination of Dex/RA has a synergistic effect on endogenous PEPCK gene expression in rat hepatocytes and H4IIE hepatoma cells. Reporter gene studies show that the gAF3 element and one of the two glucocorticoid receptor-binding elements (GR1) are most important for this effect. Chromatin immunoprecipitation assays revealed that when H4IIE cells were treated with Dex/RA, ligand-activated retinoic acid receptors (retinoic acid receptor/retinoid X receptor) and glucocorticoid receptors are recruited to this gene promoter, as are the transcription coregulators p300, CREB-binding protein, p/CIP, and SRC-1. Notably, the recruitment of p300 and RNA polymerase II to the PEPCK promoter is increased by the combined Dex/RA treatment compared with Dex or RA treatment alone. The functional importance of p300 in the Dex/RA response is illustrated by the observation that selective reduction of this coactivator, but not that of CREB-binding protein, abolishes the synergistic effect in H4IIE cells.

Peroxisome proliferator-activated receptor gamma coactivator-1alpha, as a transcription amplifier, is not essential for basal and hormone-induced phosphoenolpyruvate carboxykinase gene expression.

Phosphoenolpyruvate carboxykinase (PEPCK) catalyzes the initial step in hepatic gluconeogenesis. In the fasted state, PEPCK gene expression is activated by glucagon (via cAMP) and glucocorticoids. Peroxisome proliferator-activated receptor gamma coactivator 1alpha (PGC-1alpha) plays an important role in energy homeostasis and is considered to be a key regulator of hepatic gluconeogenesis in response to fasting. It is not clear whether PGC-1alpha is obligatory for the activation of the transcription program of gluconeogenic genes, or whether it amplifies an existing process. H4IIE hepatoma cells were used to address this key point. These cells respond appropriately to all of the hormones involved in the regulation of gluconeogenic genes, yet they are devoid of PGC-1alpha. Also, these hormone responses occur in the absence of ongoing protein synthesis, so the necessary complement of transcription factors exists in untreated cells. However, exogenous expression of PGC-1alpha in these cells does enhance basal and hormone-induced expression of the PEPCK and glucose-6-phosphatase genes. Mutational analyses of the PEPCK gene promoter reveal that one element in the PEPCK gene promoter, glucocorticoid accessory factor 3, which binds chicken ovalbumin upstream promoter-transcription factor, is of particular importance. Taken together, these data suggest that, under chronic fasting conditions, i.e. when high levels of cAMP and glucocorticoids induce PGC-1alpha expression, this coactivator markedly amplifies PEPCK gene expression and gluconeogenesis.

The Xenopus B2 factor involved in TFIIIA gene regulation is closely related to Sp1 and interacts in a complex with USF.

In the Xenopus laevis oocyte there is a million fold more transcription factor IIIA (TFIIIA) and its corresponding mRNA than in a somatic cell. These high levels of TFIIIA gene expression are achieved primarily by transcriptional regulation. The TATA box along with three positive cis-elements in the control region of the TFIIIA gene located at positions -269 to -264 (E1), -235 to -220 (E2), and -669 to -636 (E3) are required for this high level of expression in oocytes. The proteins that bind E1 and E3 of the TFIIIA gene have been identified as Xenopus USF (Xl-USF) and B3 (homolog of Vg1 RBP/VERA). In this study the B2 protein was found to bind E2 in a zinc-dependent fashion and anti-human Sp1 (but not Sp2, Sp3, nor Sp4) supershifted the B2:element 2 complex. The E2 binding protein was purified by DNA affinity chromatography. Based on supershift analysis, molecular weight estimation experiments, and purified human Sp1 DNA binding affinity tests the data strongly support the idea that the B2 protein is the Xenopus ortholog of Sp1, but not Sp2, Sp3, nor Sp4. Xl-USF binds to element 1 of the TFIIIA gene which is immediately adjacent to element 2. Coimmunoprecipitation experiments using crude whole oocyte extracts revealed that Xenopus Sp1 and USF or closely related factors are present together in a high-affinity complex. This structure contributes positively to the initiation of TFIIIA gene transcription in Xenopus oocytes.

Analysis of macroinvertebrate assemblages in relation to environmental gradients among lotic habitats of California's Central Valley.

We assessed relationships between environmental characteristics and macroinvertebrate assemblages in lotic habitats of California's Central Valley with community metric and multivariate statistical approaches. Using canonical ordination analyses, we contrasted results when assemblage structure was assessed with macroinvertebrate metrics, as suggested for use in indices of biotic integrity, or with genera abundances. Our objectives were to identify metrics or taxa diagnostic of lotic environmental stressors and compare the capacity of these approaches to detect stressors in order to suggest how they might be used to diagnose stressors. For macroinvertebrate metrics, redundancy analysis (RDA) extracted three axes correlated with channel morphology and substrates. For genera abundances, canonical correspondence analysis (CCA) extracted two axes correlated with soluble salts and with channel morphology and substrates but did not separate these gradients onto different axes. Cluster analyses identified five RDA and five CCA site groups, which exhibited differences for environmental variables, metrics, or genera abundances, and agreement between the analyses in partitioning of sites was greater than if sites were partitioned randomly. These approaches differ in their ability to detect environmental stressors, because they measure different aspects of assemblages and would be complementary in design of new metrics diagnostic of stressors.

Epigallocatechin gallate, a constituent of green tea, represses hepatic glucose production.

Herbs have been used for medicinal purposes, including the treatment of diabetes, for centuries. Plants containing flavonoids are used to treat diabetes in Indian medicine and the green tea flavonoid, epigallocatechin gallate (EGCG), is reported to have glucose-lowering effects in animals. We show here that the regulation of hepatic glucose production is decreased by EGCG. Furthermore, like insulin, EGCG increases tyrosine phosphorylation of the insulin receptor and insulin receptor substrate-1 (IRS-1), and it reduces phosphoenolpyruvate carboxykinase gene expression in a phosphoinositide 3-kinase-dependent manner. EGCG also mimics insulin by increasing phosphoinositide 3-kinase, mitogen-activated protein kinase, and p70(s6k) activity. EGCG differs from insulin, however, in that it affects several insulin-activated kinases with slower kinetics. Furthermore, EGCG regulates genes that encode gluconeogenic enzymes and protein-tyrosine phosphorylation by modulating the redox state of the cell. These results demonstrate that changes in the redox state may have beneficial effects for the treatment of diabetes and suggest a potential role for EGCG, or derivatives, as an antidiabetic agent.