Analysis of Nitrogen Loading Reductions for Wastewater Treatment Facilities and Non-Point Sources in the Great Bay Estuary Watershed

In 2009, the New Hampshire Department of Environmental Services (DES) published a proposal for numeric nutrient criteria for the Great Bay Estuary. The report found that total nitrogen concentrations in most of the estuary needed to be less than 0.3 mg N/L to prevent loss of eelgrass habitat and less than 0.45 mg N/L to prevent occurrences of low dissolved oxygen. Based on these criteria and an analysis of a compilation of data from at least seven different sources, DES concluded that 11 of the 18 subestuaries in the Great Bay Estuary were impaired for nitrogen. Under the Clean Water Act, if a water body is determined to be impaired, a study must be completed to determine the existing loads of the pollutant and the load reductions that would be needed to meet the water quality standard. Therefore, DES developed models to determine existing nitrogen loads and nitrogen loading thresholds for the subestuaries to comply with the numeric nutrient criteria. DES also evaluated the effects of different permitting scenarios for wastewater treatment facilities on nitrogen loads and the costs for wastewater treatment facility upgrades. This modeling exercise showed that: Nitrogen loads to the Great Bay, Little Bay, and the Upper Piscataqua River need to be reduced by 30 to 45 percent to attain the numeric nutrient criteria. Both wastewater treatment facilities and non-point sources will need to reduce nitrogen loads to attain the numeric nutrient criteria. The percent reduction targets for nitrogen loads only change minimally between wet and dry years. Wastewater treatment facility upgrades to remove nitrogen will be costly; however, the average cost per pound of nitrogen removed from the estuary due to wastewater facility upgrades is lower than for non-point source controls. The permitting options for some wastewater treatment facilities will be limited by requirements to not increase pollutant loads to impaired waterbodies. The numeric nutrient criteria and models used by DES are sufficiently accurate for calculating nitrogen loading thresholds for the Great Bay watershed. Additional monitoring and modeling is needed to better characterize conditions and nitrogen loading thresholds for the Lower Piscataqua River. This nitrogen loading analysis for Great Bay may provide a framework for setting nitrogen permit limits for wastewater treatment facilities and developing watershed implementation plans to reduce nitrogen loads

Staphylococcus aureus Biofilm and Planktonic cultures differentially impact gene expression, mapk phosphorylation, and cytokine production in human keratinocytes

<p>Abstract</p> <p>Background</p> <p>Many chronic diseases, such as non-healing wounds are characterized by prolonged inflammation and respond poorly to conventional treatment. Bacterial biofilms are a major impediment to wound healing. Persistent infection of the skin allows the formation of complex bacterial communities termed biofilm. Bacteria living in biofilms are phenotypically distinct from their planktonic counterparts and are orders of magnitude more resistant to antibiotics, host immune response, and environmental stress. <it>Staphylococcus aureus </it>is prevalent in cutaneous infections such as chronic wounds and is an important human pathogen.</p> <p>Results</p> <p>The impact of <it>S. aureus </it>soluble products in biofilm-conditioned medium (BCM) or in planktonic-conditioned medium (PCM) on human keratinocytes was investigated. Proteomic analysis of BCM and PCM revealed differential protein compositions with PCM containing several enzymes involved in glycolysis. Global gene expression of keratinocytes exposed to biofilm and planktonic <it>S. aureus </it>was analyzed after four hours of exposure. Gene ontology terms associated with responses to bacteria, inflammation, apoptosis, chemotaxis, and signal transduction were enriched in BCM treated keratinocytes. Several transcripts encoding cytokines were also upregulated by BCM after four hours. ELISA analysis of cytokines confirmed microarray results at four hours and revealed that after 24 hours of exposure, <it>S. aureus </it>biofilm induced sustained low level cytokine production compared to near exponential increases of cytokines in planktonic treated keratinocytes. The reduction in cytokines produced by keratinocytes exposed to biofilm was accompanied by suppressed phosphorylation of MAPKs. Chemical inhibition of MAPKs did not drastically reduce cytokine production in BCM-treated keratinocytes suggesting that the majority of cytokine production is mediated through MAPK-independent mechanisms.</p> <p>Conclusions</p> <p>Collectively the results indicate that <it>S. aureus </it>biofilms induce a distinct inflammatory response compared to their planktonic counterparts. The differential gene expression and production of inflammatory cytokines by biofilm and planktonic cultures in keratinocytes could have implications for the formation and persistence of chronic wounds. The formation of a biofilm should be considered in any study investigating host response to bacteria.</p

Redesigned and chemically-modified hammerhead ribozymes with improved activity and serum stability

BACKGROUND: Hammerhead ribozymes are RNA-based molecules which bind and cleave other RNAs specifically. As such they have potential as laboratory reagents, diagnostics and therapeutics. Despite having been extensively studied for 15 years or so, their wide application is hampered by their instability in biological media, and by the poor translation of cleavage studies on short substrates to long RNA molecules. This work describes a systematic study aimed at addressing these two issues. RESULTS: A series of hammerhead ribozyme derivatives, varying in their hybridising arm length and size of helix II, were tested in vitro for cleavage of RNA derived from the carbamoyl phosphate synthetase II gene of Plasmodium falciparum. Against a 550-nt transcript the most efficient (t(1/2 )= 26 seconds) was a miniribozyme with helix II reduced to a single G-C base pair and with twelve nucleotides in each hybridising arm. Miniribozymes of this general design were targeted to three further sites, and they demonstrated exceptional cleavage activity. A series of chemically modified derivatives was prepared and examined for cleavage activity and stability in human serum. One derivative showed a 10(3)-fold increase in serum stability and a doubling in cleavage efficiency compared to the unmodified miniribozyme. A second was almost 10(4)-fold more stable and only 7-fold less active than the unmodified parent. CONCLUSION: Hammerhead ribozyme derivatives in which helix II is reduced to a single G-C base pair cleave long RNA substrates very efficiently in vitro. Using commonly available phosphoramidites and reagents, two patterns of nucleotide substitution in this derivative were identified which conferred both good cleavage activity against long RNA targets and good stability in human serum

Towards a risk register for natural capital

1. Natural capital is essential for goods and services on which people depend. Yet pressures on the environment mean that natural capital assets are continuing to decline and degrade, putting such benefits at risk. Systematic monitoring of natural assets is a major challenge that could be both unaffordable and unmanageable without a way to focus efforts. Here we introduce a simple approach, based on the commonly used management tool of a risk register, to highlight natural assets whose condition places benefits at risk. 2. We undertake a preliminary assessment using a risk register for natural capital assets in the UK based solely on existing information. The status and trends of natural capital assets are assessed using asset–benefit relationships for ten kinds of benefits (food, fibre (timber), energy, aesthetics, freshwater (quality), recreation, clean air, wildlife, hazard protection and equable climate) across eight broad habitat types in the UK based on three dimensions of natural capital within each of the habitat types (quality, quantity and spatial configuration). We estimate the status and trends of benefits relative to societal targets using existing regulatory limits and policy commitments, and allocate scores of high, medium or low risk to asset–benefit relationships that are both subject to management and of concern. 3. The risk register approach reveals substantial gaps in knowledge about asset–benefit relationships which limit the scope and rigour of the assessment (especially for marine and urban habitats). Nevertheless, we find strong indications that certain assets (in freshwater, mountain, moors and heathland habitats) are at high risk in relation to their ability to sustain certain benefits (especially freshwater, wildlife and climate regulation). 4. Synthesis and applications. With directed data gathering, especially to monitor trends, improve metrics related to asset–benefit relationships, and improve understanding of nonlinearities and thresholds, the natural capital risk register could provide a useful tool. If updated regularly, it could direct monitoring efforts, focus research and protect and manage those natural assets where benefits are at highest risk

Late Quaternary sea-level changes of the Persian Gulf

Late Quaternary reflooding of the Persian Gulf climaxed with the mid-Holocene highstand previously variously dated between 6 and 3.4 ka. Examination of the stratigraphic and paleoenvironmental context of a mid- Holocenewhale beaching allows us to accurately constrain the timing of the transgressive, highstand and regressive phases of the mid- to late Holocene sea-level highstand in the Persian Gulf. Mid-Holocene transgression of the Gulf surpassed today's sea level by 7100–6890 cal yr BP, attaining a highstand of N1 m above current sea level shortly after 5290–4570 cal yr BP before falling back to current levels by 1440–1170 cal yr BP. The cetacean beached into an intertidal hardground pond during the transgressive phase (5300–4960 cal yr BP) with continued transgression interring the skeleton in shallow-subtidal sediments. Subsequent relative sea-level fall produced a forced regression with consequent progradation of the coastal system. These new ages refine previously reported timings for the mid- to late Holocene sea-level highstand published for other regions. By so doing, they allow us to constrain the timing of this correlatable global eustatic event more accurately

α,α-disubstituted β-amino amides eliminate Staphylococcus aureus biofilms by membrane disruption and biomass removal

Bacterial biofilms account for up to 80% of all infections and complicate successful therapies due to their intrinsic tolerance to antibiotics. Biofilms also cause serious problems in the industrial sectors, for instance due to the deterioration of metals or microbial contamination of products. Efforts are put in finding novel strategies in both avoiding and fighting biofilms. Biofilm control is achieved by killing and/or removing biofilm or preventing transition to the biofilm lifestyle. Previous research reported on the anti-biofilm potency of α,α-disubstituted β-amino amides A1, A2 and A3, which are small antimicrobial peptidomimetics with a molecular weight below 500 Dalton. In the current study it was investigated if these derivatives cause a fast disintegration of biofilm bacteria and removal of Staphylococcus aureus biofilms. One hour incubation of biofilms with all three derivatives resulted in reduced metabolic activity and membrane permeabilization in S. aureus (ATCC 25923) biofilms. Bactericidal properties of these derivatives were attributed to a direct effect on membranes of biofilm bacteria. The green fluorescence protein expressing Staphylococcus aureus strain AH2547 was cultivated in a CDC biofilm reactor and utilized for disinfectant efficacy testing of A3, following the single-tube method (American Society for Testing and Materials designation number E2871). A3 at a concentration of 90 μM acted as fast as 100 μM chlorhexidine and was equally effective. Confocal laser scanning microscopy studies showed that chlorhexidine treatment lead to fluorescence fading indicating membrane permeabilization but did not cause biomass removal. In contrast, A3 treatment caused a simultaneous biofilm fluorescence loss and biomass removal. These dual anti-biofilm properties make α,α-disubstituted β-amino amides promising scaffolds in finding new control strategies against recalcitrant biofilms

Robustness analysis of culturing perturbations on Escherichia coli colony biofilm beta-lactam and aminoglycoside antibiotic tolerance

<p>Abstract</p> <p>Background</p> <p>Biofilms are ubiquitous. For instance, the majority of medical infections are thought to involve biofilms. However even after decades of investigation, the <it>in vivo </it>efficacy of many antimicrobial strategies is still debated suggesting there is a need for better understanding of biofilm antimicrobial tolerances. The current study's goal is to characterize the robustness of biofilm antibiotic tolerance to medically and industrially relevant culturing perturbations. By definition, robust systems will return similar, predictable responses when perturbed while non-robust systems will return very different and potentially unpredictable responses. The predictability of an antibiotic tolerance response is essential to developing, testing, and employing antimicrobial strategies.</p> <p>Results</p> <p>The antibiotic tolerance of <it>Escherichia coli </it>colony biofilms was tested against beta-lactam and aminoglycoside class antibiotics. Control scenario tolerances were compared to tolerances under culturing perturbations including 1) different nutritional environments 2) different temperatures 3) interruption of cellular quorum sensing and 4) different biofilm culture ages. Here, antibiotic tolerance was defined in terms of culturable biofilm cells recovered after a twenty four hour antibiotic treatment.</p> <p>Colony biofilm antibiotic tolerances were not robust to perturbations. Altering basic culturing parameters like nutritional environment or temperature resulted in very different, non-intuitive antibiotic tolerance responses. Some minor perturbations like increasing the glucose concentration from 0.1 to 1 g/L caused a ten million fold difference in culturable cells over a twenty four hour antibiotic treatment.</p> <p>Conclusions</p> <p>The current study presents a basis for robustness analysis of biofilm antibiotic tolerance. Biofilm antibiotic tolerance can vary in unpredictable manners based on modest changes in culturing conditions. Common antimicrobial testing methods, which only consider a single culturing condition, are not desirable since slight culturing variations can lead to very different outcomes. The presented data suggest it is essential to test antimicrobial strategies over a range of culturing perturbations relevant to the targeted application. In addition, the highly dynamic antibiotic tolerance responses observed here may explain why some current antimicrobial strategies occasionally fail.</p

Ejection fraction and mortality: A nationwide register-based cohort study of 499 153 women and men

Aims: We investigated the sex-based risk of mortality across the spectrum of left ventricular ejection fraction (LVEF) in a large cohort of patients in Australia. Methods and results: Quantified levels of LVEF from 237 046 women (48.1%) and 256 109 men undergoing first-time, routine echocardiography (2000–2019) were linked to 119 232 deaths (median 5.6 years of follow-up). Overall, 17.6% of men vs. 8.3% of women had an LVEF P\u3c 0.001] in women and 1.21 (95% CI 1.05–1.39; P = 0.008) in men. In women, an LVEF of 60.0–64.9% was also associated with a HR 1.33 (95% CI 1.16–1.52; P\u3c 0.001) for cardiovascular-related mortality. These associations were most striking in women and men aged Conclusions: Among patients investigated for suspected or established cardiovascular disease, we found clinically relevant sex-based differences in the distribution and mortality associated with an LVE