Search

The US Geological Survey and Parallel Inc. conducted experimental flights with the Tarantula Hawk (T-Hawk) unmanned aircraft system (uas ) at the Dyer and Cottonwood Ranch properties located along reaches of the Platte River near Overton, Nebraska, in July 2013. We equipped the T-Hawk uas platform with a consumer-grade digital camera to collect imagery of emergent sandbars in the reaches and used photogrammetric software and surveyed control points to generate orthophotographs and digital elevation models (dems ) of the reaches. To optimize the image alignment process, we retained and/or eliminated tie points based on their relative errors and spatial resolution, whereby minimizing the total error in the project. Additionally, we collected seven transects that traversed emergent sandbars concurrently with global positioning system location data to evaluate the accuracy of the uas survey methodology. The root mean square errors for the elevation of emergent points along each transect across the dems ranged from 0.04 to 0.12 m. If adequate survey control is established, a uas combined with photogrammetry software shows promise for accurate monitoring of emergent sandbar morphology and river management activities in short (1–2 km) river reaches.

Plant pathogens such as fungi, oomycetes, viruses and bacteria infect important crops and account for significant economic losses worldwide. Therefore, it is critical to gain insights into plant-pathogen interactions at the cellular and molecular level. The outcome of the interaction between plants and pathogens greatly differs depending on the species, strains and cultivars involved as well as environmental factors, yet typically results in stress for the plant, the pathogen or both. These biotic-induced stresses can be monitored using a wide range of techniques, of which some of the most commonly used techniques are outlined in this chapter. One widely observed feature of biotic stress in plants is the generation of reactive oxygen species (ROS) such as hydrogen peroxide (H

Plant pathogens such as fungi, oomycetes, viruses and bacteria infect important crops and account for significant economic losses worldwide. Therefore, it is critical to gain insights into plant-pathogen interactions at the cellular and molecular level. The outcome of the interaction between plants and pathogens greatly differs depending on the species, strains and cultivars involved as well as environmental factors, yet typically results in stress for the plant, the pathogen or both. These biotic-induced stresses can be monitored using a wide range of techniques, of which some of the most commonly used techniques are outlined in this chapter. One widely observed feature of biotic stress in plants is the generation of reactive oxygen species (ROS) such as hydrogen peroxide (H2O2) and superoxide (O2-). We describe the quantification of hydrogen peroxide by 3,3'-diaminobenzidine (DAB) staining and luminol-based assays, and of superoxide by nitroblue tetrazolium (NBT) staining. Other techniques detailed here include measurement of callose deposition by aniline blue staining, evaluation of cell death by trypan blue staining and analysis of the loss of membrane integrity by monitoring electrolyte leakage.

In the metal hyperaccumulator plant Noccaea caerulescens, zinc may provide a defence against pathogens. However, zinc accumulation is a variable trait in this species. We hypothesize that this variability affects the outcome of interactions between metal accumulation and the various constitutive and inducible defences that N. caerulescens shares with non-accumulator plants. We compare zinc concentrations, glucosinolate concentrations and inducible stress responses, including reactive oxygen species (ROS) and cell death, in four N. caerulescens populations, and relate these to the growth of the plant pathogen Pseudomonas syringae, its zinc tolerance mutants and Pseudomonas pathogens isolated from a natural population of N. caerulescens. The populations display strikingly different combinations of defences. Where defences are successful, pathogens are limited primarily by metals, cell death or organic defences; there is evidence of population-dependent trade-offs or synergies between these. In addition, we find evidence that Pseudomonas pathogens have the capacity to overcome any of these defences, indicating that the arms race continues. These data indicate that defensive enhancement, joint effects and trade-offs between different forms of defence are all plausible explanations for the variation we observe between populations, with factors including metal availability and metal-tolerant pathogen load probably shaping the response of each population to infection.

Wildfires are a major driver of carbon stocks and ecosystem development in Canadian boreal forests, but there is little information on amounts and properties of the charcoal produced. Using data and samples available from a previous study, we determined amounts, depth distribution and chemical properties of visually-determined charcoal (>2 mm) in a boreal chronosequence in the Abitibi region of Quebec, Canada. Sites ranged from 24 to 2,355 years since fire (ysf) and originated from low- and high-severity soil burns (>5 or

Background: Pseudomonas species are adapted to evade innate immune responses and can persist at sites of relative tissue hypoxia, including the mucus-plugged airways of patients with cystic fibrosis and bronchiectasis. The ability of these bacteria to directly sense and respond to changes in local oxygen availability is in part consequent upon expression of the 2-oxoglutarate oxygenase, Pseudomonas prolyl hydroxylase (PPHD), which acts on elongation factor Tu (EF-Tu), and is homologous with the human hypoxia inducible factor (HIF) prolyl hydroxylases. We report that PPHD expression regulates the neutrophil response to acute pseudomonal infection. Methods: In vitro co-culture experiments were performed with human neutrophils and PPHD-deficient and wild-type bacteria and supernatants, with viable neutrophil counts determined by flow cytometry. In vivo consequences of infection with PPHD deficient P. aeruginosa were determined in an acute pneumonia mouse model following intra-tracheal challenge. Results: Supernatants of PPHD-deficient bacterial cultures contained higher concentrations of the phenazine exotoxin pyocyanin and induced greater acceleration of neutrophil apoptosis than wild-type PAO1 supernatants in vitro. In vivo infection with PPHD mutants compared to wild-type PAO1 controls resulted in increased levels of neutrophil apoptosis and impaired control of infection, with higher numbers of P. aeruginosa recovered from the lungs of mice infected with the PPHD-deficient strain. This resulted in an overall increase in mortality in mice infected with the PPHD-deficient strain. Conclusions: Our data show that Pseudomonas expression of its prolyl hydroxylase influences the outcome of host-pathogen interactions in vitro and in vivo, demonstrating the importance of considering how both host and pathogen adaptations to hypoxia together define outcomes of infection. Given that inhibitors for the HIF prolyl hydroxylases are in late stage trials for the treatment of anaemia and that the active sites of PPHD and human HIF prolyl hydroxylases are closely related, the results are of current clinical interest.

When plants are infected with pathogens they defend themselves via various processes. Once such process is the development of the hypersensitive response (HR), a kind of programmed cell death (PCD), in which localized cell death takes place in order to prevent pathogen spread to other part of tissue. The Arabidopsis and Pseudomonas syringae system is one of the best known examples to study the HR. Here we used the VisiSens™ oxygen-imaging system to investigate oxygen distributions in Arabidopsis leaves infected with an avirulent strain of P. syringae and undergoing the hypersensitive response. Using this method we observed a change in oxygen status at 6 h post-infection and a drop in oxygen levels at 24 h after infection.

Increasing assaults and behavioral disturbances from patients led to creation of a workgroup with representatives from Nursing, Psychiatry, Psychology, and Administration to find patient-centered ways of decreasing assault rates. The Behavior Rapid Response Team (BRRT) was created to provide early intervention and behavioral consultation for patients showing escalating anger and potential for aggression before behavior requires an emergency response. BRRT is available 24/7 and is staffed by Nursing and Psychiatry. This program was rolled out in VA Mental Health and Community Living Center units and refined prior to rollout in Acute Medical units. BRRT events and outcomes are documented and tracked. Assaults have decreased dramatically in CLC units - 63 assaults in 2013 down to 13 assaults in 2015 - and Nursing has reported increased confidence in their ability to manage behaviorally difficult patients. The BRRT consultation process and outcomes will be discussed.

Despite years of research into microbial activity at diffuse flow hydrothermal vents, the extent of microbial niche diversity in these settings is not known. To better understand the relationship between microbial activity and the associated physical and geochemical conditions, we obtained co-registered metatranscriptomic and geochemical data from a variety of different fluid regimes within the ASHES vent field on the Juan de Fuca Ridge. Microbial activity in the majority of the cool and warm fluids sampled was dominated by a population of Gammaproteobacteria (likely sulfur oxidizers) that appear to thrive in a variety of chemically distinct fluids. Only the warmest, most hydrothermally-influenced flows were dominated by active populations of canonically vent-endemic Epsilonproteobacteria. These data suggest that the Gammaproteobacteria collected during this study may be generalists, capable of thriving over a broader range of geochemical conditions than the Epsilonproteobacteria. Notably, the apparent metabolic activity of the Gammaproteobacteria—particularly carbon fixation—in the seawater found between discrete fluid flows (the intra-field water) suggests that this area within the Axial caldera is a highly productive, and previously overlooked, habitat. By extension, our findings suggest that analogous, diffuse flow fields may be similarly productive and thus constitute a very important and underappreciated aspect of deep-sea biogeochemical cycling that is occurring at the global scale.

Background and Purpose— Established risk factors do not fully identify patients at risk for recurrent stroke. The SPARCL trial (Stroke Prevention by Aggressive Reduction in Cholesterol Levels) evaluated the effect of atorvastatin on stroke risk in patients with a recent stroke or transient ischemic attack and no known coronary heart disease. This analysis explored the relationships between 13 plasma biomarkers assessed at trial enrollment and the occurrence of outcome strokes. Methods— We conducted a case–cohort study of 2176 participants; 562 had outcome strokes and 1614 were selected randomly from those without outcome strokes. Time to stroke was evaluated by Cox proportional hazards models. Results— There was no association between time to stroke and lipoprotein-associated phospholipase A 2 , monocyte chemoattractant protein-1, resistin, matrix metalloproteinase-9, N-terminal fragment of pro-B-type natriuretic peptide, soluble vascular cell adhesion molecule-1, soluble intercellular adhesion molecule-1, or soluble CD40 ligand. In adjusted analyses, osteopontin (hazard ratio per SD change, 1.362; P P =0.0107), myeloperoxidase (hazard ratio, 1.177; P =0.0022), and adiponectin (hazard ratio, 1.207; P =0.0013) were independently associated with outcome strokes. After adjustment for the Stroke Prognostic Instrument-II and treatment, osteopontin, neopterin, and myeloperoxidase remained independently associated with outcome strokes. The addition of these 3 biomarkers to Stroke Prognostic Instrument-II increased the area under the receiver operating characteristic curve by 0.023 ( P =0.015) and yielded a continuous net reclassification improvement (29.1%; P P Conclusions— Osteopontin, neopterin, and myeloperoxidase were independently associated with the risk of recurrent stroke and improved risk classification when added to a clinical risk algorithm. Clinical Trial Registration— URL: http://www.clinicaltrials.gov . Unique Identifier: NCT00147602.

This work serves as a proof-of-concept for bacterially derived SimCells (Simple Cells), which contain the cell machinery from bacteria and designed DNA (or potentially a simplified genome) to instruct the cell to carry out novel, specific tasks. SimCells represent a reprogrammable chassis without a native chromosome, which can host designed DNA to perform defined functions. In this paper, the use of Escherichia coli MC1000 ∆minD minicells as a non-reproducing chassis for SimCells was explored, as demonstrated by their ability to act as sensitive biosensors for small molecules. Highly purified minicells derived from E. coli strains containing gene circuits for biosensing were able to transduce the input signals from several small molecules (glucarate, acrylate and arabinose) into the production of green fluorescent protein (GFP). A mathematical model was developed to fit the experimental data for induction of gene expression in SimCells. The intracellular ATP level was shown to be important for SimCell function. A purification and storage protocol was developed to prepare SimCells which could retain their functions for an extended period of time. This study demonstrates that SimCells are able to perform as ‘smart bioparticles’ controlled by designed gene circuits.

The application of RNA interactome capture to plants has enabled comprehensive determination of the plant RNA-binding proteome and the identification of novel families of RNA-binding proteins (RBPs). The technique is providing insight into the evolution of the eukaryotic repertoire of RBPs and will enhance prospects for engineering RBPs to improve crop traits.

Summary One of the most fundamental questions in plant pathology is what determines whether a pathogen grows within a plant? This question is frequently studied in terms of the role of elicitors and pathogenicity factors in the triggering or overcoming of host defences. However, this focus fails to address the basic question of how the environment in host tissues acts to support or restrict pathogen growth. Efforts to understand this aspect of host–pathogen interactions are commonly confounded by several issues, including the complexity of the plant environment, the artificial nature of many experimental infection systems and the fact that the physiological properties of a pathogen growing in association with a plant can be very different from the properties of the pathogen in culture. It is also important to recognize that the phenotype and evolution of pathogen and host are inextricably linked through their interactions, such that the environment experienced by a pathogen within a host, and its phenotype within the host, is a product of both its interaction with its host and its evolutionary history, including its co-evolution with host plants. As the phenotypic properties of a pathogen within a host cannot be defined in isolation from the host, it may be appropriate to think of pathogens as having an ‘extended phenotype’ that is the product of their genotype, host interactions and population structure within the host environment. This article reflects on the challenge of defining and studying this extended phenotype, in relation to the questions posed below, and considers how knowledge of the phenotype of pathogens in the host environment could be used to improve disease control. What determines whether a pathogen grows within a plant? What aspects of pathogen biology should be considered in describing the extended phenotype of a pathogen within a host? How can we study the extended phenotype in ways that provide insights into the phenotypic properties of pathogens during natural infections?

The exclusive charmonium production process in (P) over barp annihilation with an associated pi 0 meson (p) over barp -> J/psi pi(0) is studied in the framework of QCD collinear factorization. The feasibility of measuring this reaction through the J/psi -> e(+) e(-) decay channel with the AntiProton ANnihilation at DArmstadt ((P) over bar ANDA) experiment is investigated. Simulations on signal reconstruction efficiency as well as the background rejection from various sources including the (P) over barp -> pi(+)pi(-)pi(0) and (p) over barp -> J/psi pi(0)pi(0) reactions are performed with PANDAROOT, the simulation and analysis software framework of the (P) over bar ANDA experiment. It is shown that the measurement can be done at (P) over bar ANDA with significant constraining power under the assumption of an integrated luminosity attainable in four to five months of data taking at the maximum design luminosity.

ObjectivesIn addition to hysterectomy and bilateral salpingo-oophorectomy, comprehensive surgical staging for endometrial cancer includes pelvic and para-aortic lymphadenectomy. Clarifying and addressing the morbidity from these surgical procedures is imperative. The goal of this study was to assess the prevalence of lower extremity swelling after surgery for endometrial cancer.Materials and MethodsWe performed a descriptive, cross-sectional survey study of women who underwent surgery for endometrial cancer at our institution from 2006 to 2008. Survey information included symptoms, management, and education regarding lymphedema. Demographic information such as race and education was collected in addition to clinical data such as body mass index and age.ResultsOf the 482 patients identified, 440 were determined eligible and 305 (69.3%) responded to the survey with information on lower limb swelling (LLS). Of the 108 (35%) responders who reported swelling, only 68 (22%) participants reported a diagnosis of lower limb lymphedema (LLL). The most commonly experienced symptoms among those who reported LLS were tightness, pain/tenderness, and heaviness. Among those with a diagnosis of LLL, most (60%) stated it affected their daily activities and noted exacerbating factors such as prolonged standing, heat, and walking. The most common therapies used to reduce symptoms included leg elevation (96%), compression stockings (65%), diuretics (46%), massage therapy (35%), and bandaging (25%). There was no association between LLS or LLL diagnosis and body mass index, age, race, and tobacco use. Only 8% of responders reported receiving preoperative education regarding risks for LLS and a desire for more comprehensive education was frequently noted.ConclusionsThe patient-reported incidence of LLS occurred in approximately 35% of survey participants who underwent surgery for endometrial cancer. However, only 22% reported a diagnosis of LLL. Efforts to obtain the true incidence of LLL and to develop effective educational materials and programs to improve the management of lymphedema are warranted.

The roles of 2-oxoglutarate (2OG)-dependent prolyl-hydroxylases in eukaryotes include collagen stabilization, hypoxia sensing, and translational regulation. The hypoxia-inducible factor (HIF) sensing system is conserved in animals, but not in other organisms. However, bioinformatics imply that 2OG-dependent prolyl-hydroxylases (PHDs) homologous to those acting as sensing components for the HIF system in animals occur in prokaryotes. We report cellular, biochemical, and crystallographic analyses revealing that Pseudomonas prolyl-hydroxylase domain containing protein (PPHD) contain a 2OG oxygenase related in structure and function to the animal PHDs. A Pseudomonas aeruginosa PPHD knockout mutant displays impaired growth in the presence of iron chelators and increased production of the virulence factor pyocyanin. We identify elongation factor Tu (EF-Tu) as a PPHD substrate, which undergoes prolyl-4-hydroxylation on its switch I loop. A crystal structure of PPHD reveals striking similarity to human PHD2 and a Chlamydomonas reinhardtii prolyl-4-hydroxylase. A crystal structure of PPHD complexed with intact EF-Tu reveals that major conformational changes occur in both PPHD and EF-Tu, including a >20-Å movement of the EF-Tu switch I loop. Comparison of the PPHD structures with those of HIF and collagen PHDs reveals conservation in substrate recognition despite diverse biological roles and origins. The observed changes will be useful in designing new types of 2OG oxygenase inhibitors based on various conformational states, rather than active site iron chelators, which make up most reported 2OG oxygenase inhibitors. Structurally informed phylogenetic analyses suggest that the role of prolyl-hydroxylation in human hypoxia sensing has ancient origins.

In the Canadian boreal forest, jack pine stands generally have a thin forest floor and occupy sites with coarsetextured soils and good drainage. Black spruce occurs more often on poorly drained sites and develops a thick mossdominated forest floor, but the common attribution of this development to poor quality of black spruce foliar litter has not been tested. We determined needle, twig, cone, and bark litter inputs during 10 y for black spruce and jack pine along the Boreal Forest Transect Case Study in northern Saskatchewan and Manitoba. Analysis of C, N, total phenolics, condensed tannins, and solid-state 13C NMR spectra from years 1–3 showed only small differences between species, notably higher tannins in black spruce cones. There was similarly little difference between area-based inputs, including classes of C structures determined by NMR. Condensed tannin input for black spruce was approximately twice that for jack pine, but both were in the very low range of reported values. Similar anal...

Bacteria in the diverse Pseudomonas fluorescens group include rhizosphere inhabitants known for their antifungal metabolite production and biological control of plant disease, such as Pseudomonas protegens Pf-5, and mushroom pathogens, such as Pseudomonas tolaasii. Here, we report that strain Pf-5 causes brown, sunken lesions on peeled caps of the button mushroom (Agaricus bisporus) that resemble brown blotch symptoms caused by P. tolaasii. Strain Pf-5 produces six known antifungal metabolites under the control of the GacS/GacA signal transduction system. A gacA mutant produces none of these metabolites and did not cause lesions on mushroom caps. Mutants deficient in the biosynthesis of the antifungal metabolites 2,4-diacetylphloroglucinol and pyoluteorin caused less-severe symptoms than wild-type Pf-5 on peeled mushroom caps, whereas mutants deficient in the production of lipopeptide orfamide A caused similar symptoms to wild-type Pf-5. Purified pyoluteorin and 2,4-diacetylphloroglucinol mimicked the symptoms caused by Pf-5. Both compounds were isolated from mushroom tissue inoculated with Pf-5, providing direct evidence for their in situ production by the bacterium. Although the lipopeptide tolaasin is responsible for brown blotch of mushroom caused by P. tolaasii, P. protegens Pf-5 caused brown blotch–like symptoms on peeled mushroom caps through a lipopeptide-independent mechanism involving the production of 2,4-diacetylphloroglucinol and pyoluteorin.

As low initial uptake and essentially zero later uptake limit efficacy of N fertilization for temperate conifers, we investigated factors limiting long-term tree uptake of residual 15 N-labeled fertilizer. We used a pot bioassay to assess availability of 15 N from soil sampled 10 years after fertilization of a Douglas-fir (Pseudotsuga menziesii (Mirb.) Franco) stand with 15 N-urea (200 kg N ha−1). Douglas-fir seedlings were grown for 2 years in organic (designated LFH) and mineral soil (0–10 cm) layers reconstructed from control and fertilized plots; residual fertilizer N amounted to 10 % of LHF and 5 % of MIN N. Percentage recovery of residual 15 N in seedlings was not affected by the original season of fertilization (spring vs. fall), but differed by the source of 15 N excess. LFH was a better source of residual 15 N; 12.4 % of residual LFH 15 N was taken up by seedlings and 7.6 % transferred to soil, whereas mineral soil yielded only 8.3 % of residual 15 N to seedling uptake and 2.4 % to LFH. Extractable inorganic N was 2–3 orders of magnitude higher in fallow pots. Ten-year residual fertilizer 15 N was clearly cycling between LFH and mineral soil and available to seedlings, indicating that other factors such as denitrification, leaching, and asynchrony of soil N mineralization and tree uptake limit long-term residual N fertilizer uptake in the field.

Water (brine) co-produced with oil in the Williston Basin is some of the most saline in the nation. The Prairie Pothole Region (PPR), characterized by glacial sediments and numerous wetlands, covers the northern and eastern portion of the Williston Basin. Sheridan County, Montana, lies within the PPR and has a documented history of brine contamination. Surface water and shallow groundwater in the PPR are saline and sulfate dominated while the deeper brines are much more saline and chloride dominated. A Contamination Index (CI), defined as the ratio of chloride concentration to specific conductance in a water sample, was developed by the Montana Bureau of Mines and Geology to delineate the magnitude of brine contamination in Sheridan County. Values > 0.035 indicate contamination. Recently, the U.S. Geological Survey completed a county level geographic information system (GIS)-based vulnerability assessment of brine contamination to aquatic resources in the PPR of the Williston Basin based on the age and density of oil wells, number of wetlands, and stream length per county. To validate and better define this assessment, a similar approach was applied in eastern Sheridan County at a greater level of detail (the 2.59 km2 Public Land Survey System section grid) and included surficial geology. Vulnerability assessment scores were calculated for the 780 modeled sections and these scores were divided into ten equal interval bins representing similar probabilities of contamination. Two surface water and two groundwater samples were collected from the section with the greatest acreage of Federal land in each bin. Nineteen of the forty water samples, and at least one water sample from seven of the ten selected sections, had CI values indicating contamination. Additionally, CI values generally increased with increasing vulnerability assessment score, with a stronger correlation for groundwater samples (R2 = 0.78) than surface water samples (R2 = 0.53).

In this paper, a molecular analytical approach for detecting hydrocarbonoclastic bacteria in water is suggested as a proxy measurement for tracking petroleum discharges in industrialized or pristine aquatic environments. This approach is tested for general application in cold marine regions (freezing to 5 °C). We used amplicon sequencing and qPCR to quantify 16S rRNA and GyrB genes from oleophilic bacteria in seawater samples from two different crude oil enrichments. The first experiment was conducted in a controlled environment using laboratory conditions and natural North Sea fjord seawater (NSC) at a constant temperature of 5 °C. The second was performed in the field with natural Arctic seawater (ARC) and outdoor temperature conditions from −7 °C to around 4 °C. Although the experimental conditions for NSC and ARC differed, the temporal changes in bacterial communities were comparable and reflected oil biotransformation processes. The common bacterial OTUs for NSC and ARC had the highest identity to Colwellia rossensis and Oleispira antarctica rRNA sequences and were enriched within a few days in both conditions. Other typical oil degrading bacteria such as Alcanivorax (n-alkane degrader) and Cycloclasticus (polycyclic aromatic hydrocarbons degrader) were rapidly enriched only in NSC conditions. Both the strong correlation between Oleispira SSU gene copies and oil concentration, and the specificity of the Oleispira assay suggest that this organism is a robust bioindicator for seawater contaminated by petroleum in cold water environments. Further optimization for automation of the Oleispira assay for in situ analysis with a genosensing device is underway. The assay for Colwellia quantification requires more specificity to fewer Colwellia OTUs and a well-established dose-response relationship before those taxa are used for oil tracking purposes.

Metal hyperaccumulation is an unusual trait involving the uptake and storage of high concentrations of metals in the aerial tissues of plants. A number of hypotheses have been proposed to explain the evolution of the metal hyperaccumulation trait, of which the hypothesis that accumulated metal provides a defence against herbivores or pathogens has received most attention and support. Metal hyperaccumulation requires a range of physiological adaptations that enable plants to take up, transport, sequester and tolerate high concentrations of metal. Such adaptations may confer a fitness cost, and it has been suggested that metal hyperaccumulator plants may compensate for this cost by reducing investment in other traits such as induced disease resistance. This is supported by recent work that shows that metal hyperaccumulators such as Noccaea spp. show reduced or altered production of key components of induced disease resistance. However, an alternative explanation exists, which is that the physiological adaptations involved in metal hyperaccumulation require alterations to, or compromise, functional aspects of plant defence mechanisms. Here, the evidence for trade-offs between metal hyperaccumulation and disease resistance mechanisms is reviewed, and the nature of the physiological adaptations involved in metal hyperaccumulation and their potential to impact other forms of plant defence is discussed. It is suggested that defensive trade-offs may have been key to the evolution of the metal hyperaccumulation trait, resulting in increased dependence upon the protection conferred by metals.

Purpose Medication cost is a major factor associated with increasing health care costs in the United States. Expenditures for prescription drugs in 2013 are estimated to be $283.7 billion. Closed system transfer devices are widely used for preparation of hazardous drugs. Reports indicate the Phaseal® closed system transfer device maintains sterility in vials for 7 days, suggesting the unused portion of single-use vials could be salvaged. This study was done to determine whether using a closed system transfer device to extend the beyond-use date of single-use vials of antineoplastic medications would result in a measurable cost saving. Methods A list of 25 drugs available in single-use vials, with a chemical stability of at least 48 hours, was compiled. Use of these agents was recorded during a 50-day period in April through June 2012. Use from a total of 296 vials of 21 antineoplastic agents was recorded. After allowing for the initial use of each vial, the mean potential percentage of drug waste was calculated to be 57.03%. Results Actual savings during the study period was $96,348.70. The pharmacy avoided nearly half of the potential waste and saved a mean of 29% of each vial. The cost-saving during the study period represents a $703,047.67 annual saving; which more than offsets the $106,556.55 the pharmacy spent for the Phaseal® system in 2012. Conclusion In addition to being a protective measure to reduce exposure to hazardous agents, use of the Phaseal® system results in a reduction in drug waste, and a noticeable cost saving for antineoplastic agents.

Increased sedimentation and nutrient cycle changes in Prairie Pothole Region wetlands associated with agriculture threaten the permanence and ecological functionality of these important resources. To determine the effects of land use on sedimentation and nutrient cycling, soil cores were analyzed for cesium-137 (137Cs), lead-210 (210Pb), and potassium-40 (40K) activities; textural composition; organic and inorganic carbon (C); and total nitrogen (N) from twelve wetlands surrounded by cropland, Conservation Reserve Program (CRP) lands, or native prairie uplands. Separate soil cores from nine of these wetlands were also analyzed for phosphorus (P), nitrate (NO3), and ammonium (NH4) concentrations. Wetlands surrounded by cropland had significantly greater linear sediment accretion rates than wetlands surrounded by CRP or native prairie. Linear sediment accretion rates from wetlands surrounded by cropland were 2.7 and 6 times greater than wetlands surrounded by native prairie when calculated from the initial and peak occurrence of 137Cs, respectively, and 0.15 cm y−1 (0.06 in yr−1) greater when calculated from 210Pb. Relative to wetlands surrounded by CRP, linear sediment accretion rates for wetlands surrounded by cropland were 4.4 times greater when calculated from the peak occurrence of 137Cs. No significant differences existed between the linear sediment accretion rates between wetlands surrounded by native prairie or CRP uplands. Wetlands surrounded by cropland had increased clay, P, NO3, and NH4, and decreased total C and N concentrations compared to wetlands surrounded by native prairie. Wetlands surrounded by CRP had the lowest P and NO3 concentrations and had clay, NH4, C, and N concentrations between those of cropland and native prairie wetlands. We documented increased linear sediment accretion rates and changes in the textural and chemical properties of sediments in wetlands with cultivated uplands relative to wetlands with native prairie uplands. These findings demonstrate the value of the CRP at protecting wetland catchments to reduce sedimentation.

Objectives With the use of YouTube™, this study provides results on both the demographics and public perception for a sample of videos of canine epilepsy. Methods A random sample of 150 videos was collected, the uploader description was recorded, and viewers' comments were coded according to their emotional and informational content. Results More comments were found to be sympathetic than derogatory, and there were more information-providing than information-seeking comments. The number of sympathetic comments was found to correlate positively with the number of derogatory comments, and the number of information-providing comments correlated positively with the number of information-seeking comments. Conclusion These findings shed light on the variation in perceptions of seizures in dogs, which are interestingly more sympathetic towards dogs with epilepsy than towards humans with epilepsy. This highlights the potential future use of YouTube in investigating public views as well as in informing and educating.

Targeted delivery of a therapeutic agent to a site of pathology to ameliorate disease while limiting exposure at undesired tissues is an aspirational treatment scenario. Targeting diseased kidneys for pharmacologic treatment has had limited success. We designed an approach to target an extracellular matrix protein, the fibronectin extra domain A isoform (FnEDA), which is relatively restricted in distribution to sites of tissue injury. In a mouse unilateral ureteral obstruction (UUO) model of renal fibrosis, injury induced significant upregulation of FnEDA in the obstructed kidney. Using dual variable domain Ig (DVD-Ig) technology, we constructed a molecule with a moiety to target FnEDA and a second moiety to neutralize TGF-β. After systemic injection of the bispecific TGF-β + FnEDA DVD-Ig or an FnEDA mAb, chemiluminescent detection and imaging with whole-body single-photon emission computed tomography (SPECT) revealed significantly higher levels of each molecule in the obstructed kidney than in the nonobstructed kidney, the ipsilateral kidney of sham animals, and other tissues. In comparison, a systemically administered TGF-β mAb accumulated at lower concentrations in the obstructed kidney and exhibited a more diffuse whole-body distribution. Systemic administration of the bispecific DVD-Ig or the TGF-β mAb (1–10 mg/kg) but not the FnEDA mAb attenuated the injury-induced collagen deposition detected by immunohistochemistry and elevation in Col1a1, FnEDA, and TIMP1 mRNA expression in the obstructed kidney. Overall, systemic delivery of a bispecific molecule targeting an extracellular matrix protein and delivering a TGF-β mAb resulted in a relatively focal uptake in the fibrotic kidney and reduced renal fibrosis.

Absorption is central to video game enjoyment and a major component of the development of consciousness. Games are rich sources of perceptual experiences and may facilitate absorption but transpersonal experiences are not guaranteed. Sustained absorption in the game world is tied to the development of player cognitive skills, and to the game world's perceptual environments, interactive functions, and narratives, including challenges and archetypical characters. Games are perceptually detailed, and technology advances are facilitating our entry into narrative universes of virtual fantasy and virtual reality, populated by archetypes, that are limited only by the imagination. Action games make available a rich cognitive, social, and cultural context where players may achieve sustained absorption and access higher levels of consciousness. Parallels between dreams and games, as well as reciprocal processing and behavioral transfers point to games' emotional benefits. This chapter examines digital media features that facilitate transpersonal experiences and identifies ways that makers of virtual worlds can create experiences that amplify consciousness.

Single-event upsets in the configuration memory of the 28-nm Xilinx Kintex-7 FPGA, used in the PANDA electromagnetic calorimeter, have been studied. Results from neutron and proton irradiations at energies up to 184 MeV are presented and compared with previous experimental results. In order to gain information about the energy-dependence of the single-event upset cross section, a GEANT4-based Monte Carlo simulation of upset mechanisms in nanometric silicon volumes has been developed. The results from this model are shown to agree with the experimental data for both neutrons and protons. Knowledge about the energy dependence of the cross section and of the particle flux at the location of the front-end modules in PANDA enables better estimates of the mean time between failures in the electromagnetic calorimeter. At PANDA, a total neutron flux of 1·102 cm− 2 s− 1 at the location of the front-end modules is expected at the lowest antipro-ton beam momentum and a luminosity of 1·1031 cm− 2 s− 1, leading to a predicted Mean Time Between Failures of 47±10 hours per FPGA in the electromagnetic calorimeter.

When plants are infected with pathogens they defend themselves via various processes. Once such process is the development of the hypersensitive response (HR), a kind of programmed cell death (PCD), in which localized cell death takes place in order to prevent pathogen spread to other part of tissue. The Arabidopsis and Pseudomonas syringae system is one of the best known examples to study the HR. Here we used the VisiSens™ oxygen-imaging system to investigate oxygen distributions in Arabidopsis leaves infected with an avirulent strain of P. syringae and undergoing the hypersensitive response. Using this method we observed a change in oxygen status at 6 h post-infection and a drop in oxygen levels at 24 h after infection.

CYP238A1, one of the two P450 enzymes in the genome of Pseudomonas putida KT2440, has been produced heterologously in Escherichia coli, purified, and found to bind acyclic and cyclic terpene alcohols such as farnesol, nerolidol, linalool, and terpineol. The other P450 enzyme in this organism (gene locus: PP1950) was also produced in E. coli but no substrate has been identified from a limited screen. A phthalate family oxygenase reductase (PFOR) encoded by the PP1957 gene, just downstream of the PP1955 gene for CYP238A1, accepts electrons from the reduced form of both nicotinamide adenine dinucleotide (NADH) and nicotinamide adenine dinucleotide phosphate and is able to support monooxygenase activity of CYP238A1, both in vitro and in E. coli, in which both enzymes are produced. CYP238A1 oxidizes cis- and trans-nerolidol to the 9-hydroxy product, with no evidence of attack at the olefinic double bonds. The NADH turnover rate of 170 nmol(nmol-P450)⁻¹ Min⁻¹ for CYP238A1 with cis-nerolidol as substrate at a PP1957:CYP238A1 concentration ratio of 8:1 suggests that this PFOR could function as the physiological redox partner for CYP238A1. The physiological role of CYP238A1 may be related to the PP1955 gene being part of an island/cluster of inducible genes associated with energy metabolism and response to xenobiotics.

The interactions between microorganisms driven by substrate metabolism and energy flow are important to shape diversity, abundance, and structure of a microbial community. Single cell technologies are useful tools for dissecting the functions of individual members and their interactions in microbial communities. Here, we developed a novel Raman stable isotope probing (Raman-SIP), which uses Raman microspectroscopy coupled with reverse and D

In environmental microbiology, the most commonly used methods of bacterial DNA transfer are conjugation and electroporation. However, conjugation requires physical contact and cell-pilus-cell interactions; electroporation requires low-ionic strength medium and high voltage. These limitations have hampered broad applications of bacterial DNA delivery. We have employed a standard low frequency 40 kHz ultrasound bath to successfully transfer plasmid pBBR1MCS2 into Pseudomonas putida UWC1, Escherichia coli DH5alpha and Pseudomonas fluorescens SBW25 with high efficiency. Under optimal conditions: ultrasound exposure time of 10 s, 50 mM CaCl(2), temperature of 22 degrees C, plasmid concentration of 0.8 ng/microl, P. putida UWC1 cell concentration of 2.5 x 10(9) CFU (colony forming unit)/ml and reaction volume of 500 microl, the efficiency of ultrasound DNA delivery (UDD) was 9.8 +/- 2.3 x 10(-6) transformants per cell, which was nine times more efficient than conjugation, and even four times greater than electroporation. We have also transferred pBBR1MCS2 into E. coli DH5alpha and P. fluorescens SBW25 with efficiencies of 1.16 +/- 0.13 x 10(-6) and 4.33 +/- 0.78 x 10(-6) transformants per cell, respectively. Low frequency UDD can be readily scaled up, allowing for the application of UDD not only in laboratory conditions but also on an industrial scale.

Comparison of the genome sequences of three Pseudomonas fluorescens strains reveals a heterogeneity reminiscent of a species complex rather than a single species, Background Pseudomonas fluorescens are common soil bacteria that can improve plant health through nutrient cycling, pathogen antagonism and induction of plant defenses. The genome sequences of strains SBW25 and Pf0-1 were determined and compared to each other and with P. fluorescens Pf-5. A functional genomic in vivo expression technology (IVET) screen provided insight into genes used by P. fluorescens in its natural environment and an improved understanding of the ecological significance of diversity within this species. Results Comparisons of three P. fluorescens genomes (SBW25, Pf0-1, Pf-5) revealed considerable divergence: 61% of genes are shared, the majority located near the replication origin. Phylogenetic and average amino acid identity analyses showed a low overall relationship. A functional screen of SBW25 defined 125 plant-induced genes including a range of functions specific to the plant environment. Orthologues of 83 of these exist in Pf0-1 and Pf-5, with 73 shared by both strains. The P. fluorescens genomes carry numerous complex repetitive DNA sequences, some resembling Miniature Inverted-repeat Transposable Elements (MITEs). In SBW25, repeat density and distribution revealed 'repeat deserts' lacking repeats, covering approximately 40% of the genome. Conclusions P. fluorescens genomes are highly diverse. Strain-specific regions around the replication terminus suggest genome compartmentalization. The genomic heterogeneity among the three strains is reminiscent of a species complex rather than a single species. That 42% of plant-inducible genes were not shared by all strains reinforces this conclusion and shows that ecological success requires specialized and core functions. The diversity also indicates the significant size of genetic information within the Pseudomonas pan genome.

Metals play essential roles in many biological processes but are toxic when present in excess. This makes their transport and homoeostatic control of particular importance to living organisms. Within the context of plant-pathogen interactions the availability and toxicity of transition metals can have a substantial impact on disease development. Metals are essential for defensive generation of reactive oxygen species and other plant defences and can be used directly to limit pathogen growth. Metal-based antimicrobials are used in agriculture to control plant disease, and there is increasing evidence that metal hyperaccumulating plants use accumulated metal to limit pathogen growth. Pathogens and hosts compete for available metals, with plants possessing mechanisms to withhold essential metals from invading microbes. Pathogens, meanwhile, use low-metal conditions as a signal to recognise and respond to the host environment. Consequently, metal-sensing systems such as fur (iron) and zur (zinc) regulate the expression of pathogenicity and virulence genes; and pathogens have developed sophisticated strategies to acquire metal during growth in plant tissues, including the production of multiple siderophores. This review explores the impact of transition metals on the processes that determine the outcome of bacterial infection in plants, with a particular emphasis on zinc, iron and copper. This review discusses the pivotal role of transition metals such as zinc, copper and iron in determining the outcome of plant-pathogen interactions, both as essential mineral nutrients for plant and pathogen, and as anti-microbial agents that inhibit pathogen growth. © 2012 Federation of European Microbiological Societies. Published by Blackwell Publishing Ltd. All rights reserved.

Pseudomonas fluorescens SBW25 is a Gram-negative bacterium that grows in close association with plants. In common with a broad range of functionally similar bacteria it plays an important role in the turnover of organic matter and certain isolates can promote plant growth. Despite its environmental significance, the causes of its ecological success are poorly understood. Here we describe the development and application of a simple promoter trapping strategy (IVET) to identify P. fluorescens SBW25 genes showing elevated levels of expression in the sugar beet rhizosphere. A total of 25 rhizosphere-induced (rhi) fusions are reported with predicted roles in nutrient acquisition, stress responses, biosynthesis of phytohormones and antibiotics. One rhi fusion is to wss, an operon encoding an acetylated cellulose polymer. A mutant carrying a defective wss locus was competitively compromised (relative to the wild type) in the rhizosphere and in the phyllosphere, but not in bulk soil. The rhizosphere-induced wss locus therefore contributes to the ecological performance of SBW25 in the plant environment and supports our conjecture that genes inactive in the laboratory environment, but active in the wild, are likely to be determinants of fitness in natural environments.

Effective gene trapping and screening requires sensory and regulatory compatibility of both host and exogenous systems. The naturally competent bacterium Acinetobacter baylyi ADP1 is able to efficiently take up and integrate exogenous DNA into the chromosome, making it an attractive host system for a wide range of metagenomic applications. To test the ability of A. baylyi ADP1 to express the XylR-regulated Pu promoter from Pseudomonas putida mt-2, we have constructed and examined an A. baylyi ADP1 strain, ADPWH-Pu-lux-xylR. The Pu promoter in ADPWH-Pu-lux-xylR was specifically induced by toluene, m-, p- and o-xylene. The substrate-induced Pu promoter was highly dependent on the growth medium: it was repressed in rich media until stationary phase, but was immediately induced in minimal medium with glucose as the sole carbon source (MMG). However, the Pu promoter was repressed in MMG when it was supplemented with 5 g l(-1) yeast extract. Further investigation showed that the Pu promoter in MMG was repressed by 0.5 g l(-1) aspartic acid or asparagine, but not repressed by glutamine. Changing the carbon/nitrogen ratios by addition of ammonia did not significantly affect the Pu promoter activity but addition of nitrate did. These results show that A. baylyi ADP1 reproduced characteristics of the XylR-regulated Pu promoter observed in its original host. It demonstrates that A. baylyi could provide an excellent genetic host for a wide range of functional metagenomic applications.

The Pseudomonas syringae pathovars are composed of host-specific plant pathogens that characteristically elicit the defense-associated hypersensitive response (HR) in nonhost plants. P. s. pv. syringae 61 secretes an HR elicitor, harpinPss (HrpZPss), in a hrp-dependent manner. An internal fragment of the P. s. pv. syringae 61 hrpZ gene was used to clone the hrpZ locus from P. s. pv. glycinea race 4 (bacterial blight of soybean) and P. s. pv. tomato DC3000 (bacterial speck of tomato). DNA sequence analysis revealed that hrpZ is the second ORF in a polycistronic operon. The amino acid sequence identities of HrpZPss/HrpZPsg and HrpZPss/HrpZPst, were 79 and 63%, respectively. Although none of the HrpZ proteins showed significant overall sequence similarity with other known proteins, HrpZPst contained a 24-amino acid sequence that is homologous with a region of the PopA1 elicitor protein of the tomato pathogen, Pseudomonas solanacearum GMI1000. hrpA, the upstream ORF, was highly divergent: The amino acid sequence identities of HrpAPss/HrpApsg and HrpAPss/HrpAPst were 91 and 28%, respectively, and no HrpA sequence showed similarity to known proteins. In contrast, the predicted products of the downstream ORFs in P. s. pv. syringae and P. s. pv. tomato, hrpB, hrpC, hrpD, and hrpE showed varying levels of similarity to those of yscI, yscJ, yscK, and yscL. These are colinearly arranged genes in the virC locus of Yersinia spp., which are involved in the secretion of the Yop virulence proteins via the type III pathway. The similarity of the Ysc proteins was generally stronger in comparisons with the P. s. pv. tomato Hrp proteins. The HrpZ proteins were purified by heat denaturation of contaminating proteins followed by ammonium sulfate fractionation, hydrophobic chromatography, and gel electrophoresis. All three HrpZ proteins elicited the HR in tomato, whereas none of them elicited significant necrosis in soybean. The results indicate that HrpZ is encoded in an operon containing some of the genes involved in its own secretion and suggest that HrpZ structure does not directly determine bacterial host range.

Pseudomonas syringae and Erwinia chrysanthemi attack plants in seemingly dissimilar ways. E. chrysanthemi strains secrete multiple plant tissue-disintegrating pectic enzymes and cause extensive necrosis and maceration in a wide range of hosts. P. syringae strains possess considerable host specificity, have little pectolytic capacity, generally do not macerate host tissues, and cause only delayed necrosis in their hosts. Nevertheless, both pathogens possess hip genes, and they produce harpins - protein elicitors of the hypersensitive response (HR).

The bacteriovorous nematode Caenorhabditis elegans has been used to investigate many aspects of animal biology, including interactions with pathogenic bacteria. However, studies examining C. elegans interactions with bacteria isolated from environments in which it is found naturally are relatively scarce. C. elegans is frequently associated with cultivation of the edible mushroom Agaricus bisporus, and has been reported to increase the severity of bacterial blotch of mushrooms, a disease caused by bacteria from the Pseudomonas fluorescens complex. We observed that pseudomonads isolated from mushroom farms showed differential resistance to nematode predation. Under nutrient poor conditions, in which most pseudomonads were consumed, the mushroom pathogenic isolate P. fluorescens NZI7 was able to repel C. elegans without causing nematode death. A draft genome sequence of NZI7 showed it to be related to the biocontrol strain P. protegens Pf-5. To identify the genetic basis of nematode repellence in NZI7, we developed a grid-based screen for mutants that lacked the ability to repel C. elegans. The mutants isolated in this screen included strains with insertions in the global regulator GacS and in a previously undescribed GacS-regulated gene cluster, 'EDB' ('edible'). Our results suggest that the product of the EDB cluster is a poorly diffusible or cell-associated factor that acts together with other features of NZI7 to provide a novel mechanism to deter nematode grazing. As nematodes interact with NZI7 colonies before being repelled, the EDB factor may enable NZI7 to come into contact with and be disseminated by C. elegans without being subject to intensive predation. © 2013 International Society for Microbial Ecology.

Mutations in the five hrp and hrc genes in the hrpC operon of the phytopathogen Pseudomonas syringae pv. syringae 61 have different effects on bacterial interactions with host and nonhost plants. The hrcC gene within the hrpC operon encodes an outer membrane component of the Hrp secretion system that is conserved in all type III protein secretion systems and is required for most pathogenic phenotypes and for secretion of the HrpZ harpin to the bacterial milieu. The other four genes (in order), hrpF , hrpG , ( hrcC ), hrpT , and hrpV , appear to be unique to the group I hrp clusters found in certain phytopathogens (e.g., P. syringae and Erwinia amylovora ) and are less well understood. We initiated an examination of their role in Hrp regulation and secretion by determining the effects of functionally nonpolar nptII cartridge insertions in each gene on the production and secretion of HrpZ, as determined by immunoblot analysis of cell fractions. P. syringae pv. syringae 61 hrpF , hrpG , and hrpT mutants were unable to secrete HrpZ, whereas the hrpV mutant overproduced and secreted the protein. This suggested that HrpV is a negative regulator of HrpZ production. Further immunoblot assays showed that the hrpV mutant produced higher levels of proteins encoded by all three of the major hrp operons tested—HrcJ ( hrpZ operon), HrcC ( hrpC operon), and HrcQ B ( hrpU operon)—and that constitutive expression of hrpV in trans abolished the production of each of these proteins. To determine the hierarchy of HrpV regulation in the P. syringae pv. syringae 61 positive regulatory cascade, which is composed of HrpRS (proteins homologous with ς 54 -dependent promoter-enhancer-binding proteins) and HrpL (alternate sigma factor), we tested the ability of constitutively expressed hrpV to repress the activation of HrcJ production that normally accompanies constitutive expression of hrpL or hrpRS . No repression was observed, indicating that HrpV acts upstream of HrpRS in the cascade. The effect of HrpV levels on transcription of the hrpZ operon was determined by monitoring the levels of β-glucuronidase produced by a hrpA ′:: uidA transcriptional fusion plasmid in different P. syringae pv. syringae 61 strains. The hrpV mutant produced higher levels of β-glucuronidase than the wild type, a hrcU (type III secretion) mutant produced the same level as the wild type, and the strain constitutively expressing hrpV in trans produced low levels equivalent to that of a hrpS mutant. These results suggest that HrpF, HrpG, and HrpT are all components of the type III protein secretion system whereas HrpV is a negative regulator of transcription of the Hrp regulon.

In P. syringae, the co-ordinated regulation of different systems required for pathogenicity and virulence seems logical but has not been established. This question was addressed in the present study by analysing production of the phytotoxin coronatine (COR) in defined hrp/hrc mutants of P. syringae pv. tomato DC3000. COR was produced in vitro by mutants of DC3000 defective in hrcC, which encodes an outer-membrane protein required for type III-mediated secretion. When inoculated in plants, hrcC mutants produced chlorotic regions indicative of COR production, but lacked the necrotic lesions produced by the wild-type DC3000. Furthermore, a DC3000 mutant containing a polar mutation in hrcC, which inactivates hrcC, hrpT and hrpV, produced significantly higher amounts of COR than the wild-type strain in vitro. This mutant was able to produce COR earlier and at lower cell densities than the wild-type. The results indicate that the hrp/hrc secretion system is not required for COR production, but mutations in this system may have regulatory effects on the production of virulence factors such as COR.

Summary Cell sorting coupled with single‐cell genomics is a powerful tool to circumvent cultivation of microorganisms and reveal microbial ‘dark matter’. Single‐cell Raman spectra (SCRSs) are label‐free biochemical ‘fingerprints’ of individual cells, which can link the sorted cells to their phenotypic information and ecological functions. We employed a novel Raman‐activated cell ejection (RACE) approach to sort single bacterial cells from a water sample in the Red Sea based on SCRS. Carotenoids are highly diverse pigments and play an important role in phototrophic bacteria, giving strong and distinctive Raman spectra. Here, we showed that individual carotenoid‐containing cells from a Red Sea sample were isolated based on the characteristic SCRS. RACE‐based single‐cell genomics revealed putative novel functional genes related to carotenoid and isoprenoid biosynthesis, as well as previously unknown phototrophic microorganisms including an unculturable Cyanobacteria spp. The potential of Raman sorting coupled to single‐cell genomics has been demonstrated.

Extended Abstract The current lack of preventive and treatment measures available against bacterial diseases affecting crops is an important problem in agriculture. The use of antibiotics and copper compounds is restricted and can cause bacterial resistance and negative impacts on the environment [1, 2]. Nevertheless, other successful treatment methods have not been fully established. Promising natural compounds exist which have been proven to have strong antimicrobial properties, such as essential oils, and could be used to target bacterial phytopathogens. However, the volatile nature of these compounds limits their application and use in agriculture and a suitable delivery method is extremely desirable. The aim of this study is to evaluate the use of mesoporous silica nanoparticles (MSNPs) to encapsulate essential oils and be used as a preventive and control measure against Pseudomonas syringae pv. pisi, the causative agent of pea bacterial blight [3]. MSNPs allow for a targeted and controlled delivery of the essential oil in the presence of the bacterial pathogen. Essential oils are an environmentally-friendly and safe alternative to antibiotics and other antibacterial agents and the use of nanoparticles as a delivery vehicle allows the oils to be maintained in their active form for long periods of time until their release is needed. MSNPs can protect the loaded biocide from evaporation or degradation while improving their stability and miscibility in aqueous solutions. Additionally, silica is a safe, biodegradable and biocompatible material [4] that is naturally present in plant systems and has been shown to have mainly positive effects on plant growth and development [5, 6, 7, 8]. The present study evaluated the effect of 41 essential oils against P. syringae pv. pisi and compared it to the effect on two other bacterial strains; Pectobacterium carotovorum and Pseudomonas fluorescens. The effect was shown to be strainspecific and one of the most effective oils against P. syringae pv. pisi, ajwain oil, was selected for encapsulation into the MSNPs, which were then incorporated into an alginate seed coating to treat the seeds prior to sowing. Using a seed treatment as a biocide application method reduces the quantity of antimicrobials needed, as well as the amount of nanoparticles released to the environment, when compared to other methods such as soil or foliar applications. This decreases the risk of microbial resistance to biocides while increasing the safety and efficacy of the product. The effect of different alginate formulations on the plant’s germination, growth and development was studied to assess the influence of MSNPs and the use of this technology as a seed treatment against seed-borne pathogens. Peas coated with loaded MSNPs appeared to better resist infection and grow taller than control seeds. Further experiments will be performed to optimise the loading of the MSNPs and maximise the killing efficacy against the pathogens. Additionally, the applications of this study can be extended to both other pathogens as well as other host plants. To our knowledge, this is the first report of MSNPs loaded with a natural biocide and incorporated into an alginate seed coating to protect peas from bacterial phytopathogen Pseudomonas syringae pv pisi. The development of novel and safe biocide delivery methods is essential to optimise food production and achieve a sustainable agriculture in the future.

A 74-year-old man presented for evaluation after discovery of a left bladder-wall tumor. He underwent transurethral resection of bladder tumor (TURBT) operation for treatment of low-grade, Ta urothelial cancer of the bladder. The patient developed recurrent disease and returned to the operating room for repeat TURBT, circumcision, and administration of intravesical mitomycin C. The patient developed balanitis xerotica obliterans 4 years post-circumcision, requiring self-dilation with a catheter. He subsequently developed 3 consecutive episodes of left-sided pyelonephritis. Further investigation with voiding cystourethrogram (VCUG) revealed Grade 3, left-sided vesicoureteral reflux (VUR). Due to existing comorbidities, the patient elected treatment with endoscopic dextranomer/hyaluronic acid injection. A post-operative VCUG demonstrated complete resolution of left-sided VUR. This patient has remained symptom free for 8 months post-injection, with no episodes of pyelonephritis.