Effects of oil sands process water mixtures on the mayfly Hexagenia and field-collected aquatic macroinvertebrate communities.

Extraction of Canada's oil sands has created 1 billion m3 of tailings, which are stored in on-site tailings ponds. Due to limited storage capacity, the planned release of tailings into the surrounding environment may be required. This represents an environmental management challenge, as the tailings contain contaminants that are known toxins to aquatic communities. Of particular concern are naphthenic acids and their metallic counterparts, as they are the principal toxic components of tailings, are relatively soluble, and are persistent in aquatic environments. This study examines the acute toxicity of environmentally relevant 10:1 mixtures of two process water components: naphthenic acid and sodium naphthenate. We assess the effects of these simplified oil sands process water (OSPW) mixtures under planned and unplanned tailings release scenarios, using traditional and cutting-edge bioindicators for aquatic invertebrate taxa. We found that safe concentrations for mayflies and other aquatic macroinvertebrates were less than 1 mg/l, as no mayfly taxa survived repeated exposure to this dose in either the 48-h or 72-h acute toxicity test. In the 72-h test, no mayflies survived treatment levels greater than 0.5 mg sodium naphthenate/l. In the mesocosm study, even a 90% dilution of the OSPW mixture was not sufficient to protect sensitive macroinvertebrate communities. The results of this study highlight the potential environmental damage that will occur if OSPW is not carefully managed. This information will aid with the development of a management plan for oil sands tailings ponds, which will provide insight into the potential for process water release into the surrounding environment while conserving unique ecosystems downstream of development in the oil sands region.

Driving and Patients With Dementia.

Driving is a symbol of autonomy and independence, eagerly awaited during adolescence, cherished during adulthood and reluctantly rescinded during old age. It is nevertheless an individual's privilege, not right, especially as driving may affect other drivers and pedestrians on the road. It is therefore not only the individual patient who is at stake but essentially the entire community. In this case scenario, we describe the situation that arose when a patient with multi-infarct dementia wanted to go for a drive and his son and grandson tried to convince him that he could no longer drive. What went wrong in the caregivers/patient interaction is presented. The futility of arguing with patients who have dementia is highlighted as well as the suspiciousness it may generate. Alternate actions that can be useful to avoid/avert the situation from escalating and having a catastrophic ending are discussed. Testing/evaluating patients with dementia for fitness to drive is also reviewed and a list of select resources is included.

Agnosia Interferes With Daily Hygiene in Patients With Dementia.

Patients with dementia, particularly Alzheimer's disease, may not recognize that their clothes are dirty. They may see the food stains and discoloration of the clothes and yet because of their agnosia are unable to integrate these observations and deduce that their clothes are dirty and need to be changed. They will, therefore, resist attempts to get them to change clothes, especially if these clothes happen to be their favorite ones. This often causes caregivers to become frustrated, especially, if it represents a change in the patient's previous habits of only wearing clean clothes. In this case study, we present a 72-year-old woman with moderate Alzheimer's disease who lives with her daughter, who adamantly refuses to change the clothes she has been wearing for a few days and which are now clearly dirty. We report the interaction, highlight what went wrong in the patient-daughter interaction, and discuss how the catastrophic ending could have been avoided or averted.

Visual Hallucinations and Paranoid Delusions.

Visual well-formed hallucinations, fluctuations in the level of cognition, and alertness and extrapyramidal signs are core features of dementia with Lewy bodies. Some patients realize that what they are seeing or hearing are just hallucinations and learn to accept them. Others, however experience these hallucinations as quite real and cannot be dissuaded from the firm belief that they are. In fact, efforts to dissuade them often serve only to confirm the often associated paranoid delusions and this may lead to a catastrophic ending. Hence, it is best not to contradict the patient. Instead, attempts should be made to distract the patient and change the focus of her or his attention. In this case scenario, we present a 68-year-old man who has been diagnosed with dementia with Lewy bodies. He lives with his daughter. He has visual hallucinations and paranoid delusions that worsen at night: He thinks there are people outside the house plotting to kill him. We discuss what went wrong in the patient/caregiver interaction and how the catastrophic ending could have been avoided or averted.

Synthesis and synchrotron X-ray characterization of two 2D Hoffman related compounds [Ni(p-Xylylenediamine)nNi(CN)4] and [Ni(p-tetrafluoroxylylenediamine)nNi(CN)4].

Synchrotron X-ray single crystal structure determination of two 2D Hofmann-related compounds, [Ni(p-Xylyenediamine)n-tetracyanonickelate] (abbreviated as Ni-pXdam) and [Ni(tetrafluoro-p-Xylyenediamine)n-tetracyanonickelate] (abbreviated as Ni-pXdamF4), have been conducted. Both the pXdam and pXdamF4 ligands contain two short chains of -CH2NH2 at the para-positions of a phenyl ring. These flexible chains link the 6-fold coordinated Ni2 sites throughout the network. In Ni-pXdam, the closed-2D network of [Ni-(CN-Ni1/4-)4]∞ is broken into 1D chains, leaving the C≡N groups at the trans-positions of the Ni(CN)4 moiety unbridged. The resulting 1D chains [(trans-)-NC-Ni(CN)2-CN-Ni-]∞ runs along the [010] direction of the unit cell. The pXdam ligands bridge in pair between the Ni atoms of the adjacent chains. The catenation structure of [Ni{(pXdam)}]∞ could be referred to as double -1D. In Ni-pXdamF4, the -CH2NH2 ligands connect the neighboring chains via the 6-fold Ni2 site. Surrounding the 4-fold Ni1 site, the two trans terminal C≡N groups were replaced by the Lewis base NH3 during the synthesis process, therefore preventing the propagation of the 2D net to form a 3D network. Computed pore volume of both compounds indicated that there is not sufficient space in the structure to accommodate gas molecules. In both compounds, hydrogen bonds were found, and solvent of crystallization was absent due to the limited free space in the structure.

Development of environmental thresholds for streams in agricultural watersheds.

Global increases in consumption of chemical nutrients, application of pesticides, and water withdrawal to enhance agricultural yield have resulted in degraded water quality and reduced water availability. Efforts to safeguard or improve environmental conditions of agroecosystems have usually focused on managing on-farm activities to reduce materials loss and conserve habitat. Another management measure for improving environmental quality is adoption of environmental performance standards (also called outcome-based standards). This special collection of six papers presents the results of four years of research to devise scientifically credible approaches for setting environmental performance standards to protect water quantity and quality in Canadian agriculturally dominated watersheds. The research, conducted as part of Canada's National Agri-Environmental Standards Initiative, aimed to identify Ideal Performance Standards (the desired environmental state needed to maintain ecosystem health) and Achievable Performance Standards (the environmental conditions achievable using currently available and recommended best available processes and technologies). Overviews of the papers, gaps in knowledge, and future research directions are presented. As humans, livestock, and wildlife (both terrestrial and aquatic) experience greater pressures to share the same limited water resources, innovative research is needed that incorporates a landscape perspective, economics, farm practices, and ecology to advance the development and application of tools for protecting water resources in agricultural watersheds.

Application of nitrogen and phosphorus criteria for streams in agricultural landscapes.

Efforts to control eutrophication of water resources in agriculturally dominated ecosystems have focused on managing on-farm activities to reduce nutrient loss; however, another management measure for improving water quality is adoption of environmental performance criteria (or 'outcome-based standards'). Here, we review approaches for setting environmental quality criteria for nutrients, summarize approaches developed in Canada for setting 'ideal' and 'achievable' nutrient criteria for streams in agricultural watersheds, and consider how such criteria could be applied. As part of a 'National Agri-Environmental Standards Initiative', the Government of Canada committed to the development of non-regulatory environmental performance standards that establish total P (TP) and total N (TN) concentrations to protect ecological condition of agricultural streams. Application of four approaches for defining ideal standards using only chemistry data resulted in values for TP and TN spanning a relatively narrow range of concentrations within a given ecoregion. Cross-calibration of these chemically derived standards with information on biological condition resulted in recommendations for TP and TN that would likely protect aquatic life from adverse effects of eutrophication. Non-point source water quality modelling was then conducted in a specific watershed to estimate achievable standards, i.e. chemical conditions that could be attained using currently available and recommended management practices. Our research showed that, taken together, short-term achievable standards and ultimate ideal standards could be used to set policy targets that should, if realized, lower N and P concentrations in Canadian agricultural streams and improve biotic condition.

Structural and functional responses of benthic invertebrates to imidacloprid in outdoor stream mesocosms.

Structural and functional responses of a benthic macroinvertebrate assemblage to pulses of the insecticide imidacloprid were assessed in outdoor stream mesocosms. Imidacloprid pulses reduced invertebrate abundance and community diversity in imidacloprid-dosed streams compared to control streams. These results correlated well with effects of imidacloprid on leaf litter decomposition and feeding rates of Pteronarcys comstocki, a stonefly, in artificial streams. Reductions in oxygen consumption of stoneflies exposed to imidacloprid were also observed in laboratory experiments. Our findings suggest that leaf litter degradation and single species responses can be sensitive ecotoxicological endpoints that can be used as early warning indicators and biomonitoring tools for pesticide contamination. The data generated illustrates the value of mesocosm experiments in environmental assessment and how the consideration of functional and structural endpoints of natural communities together with in situ single species bioassays can improve the evaluation and prediction of pesticide effects on stream ecosystems.

Eutrophication of agricultural streams: defining nutrient concentrations to protect ecological condition.

Inputs of nutrients (phosphorus, P, and nitrogen, N) to coastal and fresh waters can accelerate eutrophication, resulting in excessive aquatic plant growth, depletion of oxygen, and deleterious changes in abundance and diversity of organisms. Using long-term (approximately 1995-2005) monitoring data from agriculturally-dominated watersheds in southern Ontario and Quebec, Canada, we developed and tested several approaches for setting targets for N and P. Our research showed that it is possible to set scientifically-credible targets for total P and total N to protect ecological condition of streams in agricultural landscapes, and define achievable targets attainable following adoption of beneficial management practices.

Effects of metal mining effluent on Atlantic salmon (Salmo salar) and slimy sculpin (Cottus cognatus): using artificial streams to assess existing effects and predict future consequences.

In the summer of 2000, the effects of metal mine discharge on fish growth and exercise performance were assessed at a Zn-Pb-Cu mine in New Brunswick, Canada. Juvenile Atlantic salmon (Salmo salar) were exposed to 0%, 20%, and 80% treated metal mine effluent in a mobile, fish-only artificial stream system. Fish were fed commercial salmon pellets throughout the study. Young-of-the-year slimy sculpins (Cottus cognatus) were exposed to the same treatments in a multitrophic level, modular artificial stream system or mesocosm, in which the fish were dependent on seeded algae and invertebrates for nutrition. Treatment concentrations were chosen to represent existing discharge dilutions (80%) and a scenario of reduced effluent discharge (20%) as predicted upon mine closure (scheduled for 2008). Al, Ba, B, Fe, Mn, Sr, Tl, Ti, and Zn increased in a concentration-dependent fashion across the three treatments. Salmon body burdens of Ba, Cd, Li, Cu, Mn, Se, Sr, and Zn were increased in the 80% treatment, while Tl increased across all treatment levels. Mortalities and depressions in growth in both fish species paralleled treatment concentrations (80%>20%>0%). Salmon liver weight was significantly greater in fish exposed to 20% and 80% effluent in a concentration-dependent fashion. Exercise performance in fish, as assessed by the ability to recover from forced exercise, showed little effect of treatment. The contamination of the receiving environment by mine discharges has led to loss of fish, making it impossible to study the system in situ. However, the use of the artificial stream systems enabled us to assess effects of present conditions on fish, as well as the potential impacts of mine reclamation. The 20% discharge predicted following mine reclamation is potentially favourable for the reinstitution of native fishes into the system.

Toxicant interactions with food algae: a missing link between laboratory and field effects?

Algae fed to invertebrate subjects of chronic toxicity testing are cultured without exposure to test substances. This approach may reduce the ability of bioassays to predict field effects because it assumes that bioconcentration is the only important uptake route, and that an interaction between toxicant and algae does not occur or is not relevant to the effect of the toxicant on test animals. The research presented in this paper focuses on the effects of a bleached kraft mill effluent (BKME) on algae used as food for test animals and the possible consequences of this exposure to bioassay results. The experiment consisted of exposing cultures of a pennate diatom, Navicula, to a range (0-7%) of BKME concentrations for 15 days. Final biomass (measured as chlorophyll a and ash free dry mass) was significantly greater in cultures exposed to 5% and 7% BKME. The carbon-to-nitrogen ratio was significantly higher in diatom cultures exposed to 7% BKME, and total lipid content ranged from 11.7% in the control to 15.8% in the 7% treatment. BKME exposure also increased bacterial content and altered the elemental composition (particularly strontium, barium, iron, and cobalt) of Navicula relative to control cultures. Because changes in food abundance and food quality (e.g., dietary lipids, carbohydrates, proteins) are known to modify toxicity and because contaminant uptake can occur through ingestion, exposing algal food supplies to toxicants would allow chronic bioassays to better simulate field conditions. This approach would be of value in situations where bioassays are intended to predict field effects rather than to compare the toxic potential of effluent samples. Although culturing food algae under exposure to contaminants poses methodological challenges, this approach may serve to enhance the predictive ability of chronic bioassays.

Induction of antigen-specific CD8+ T cells, T helper cells, and protective levels of antibody in humans by particle-mediated administration of a hepatitis B virus DNA vaccine.

A DNA vaccine against the hepatitis B virus (HBV) was evaluated for safety and induction of immune responses in 12 healthy, hepatitis-naïve human volunteers using the needle-free PowderJect system to deliver gold particles coated with DNA directly into cells of the skin. Three groups of four volunteers received three administrations of DNA encoding the surface antigen of HBV at one of the three dose levels (1, 2, or 4 microg). The vaccine was safe and well tolerated, causing only transient and mild to moderate responses at the site of administration. HBV-specific antibody and both CD4+ and CD8+ T cell responses were measured before and after each immunization. All the volunteers developed protective antibody responses of at least 10 mIU/ml. In volunteers who were positive for the HLA class I A2 allele, the vaccine also induced antigen-specific CD8+ T cells that bound HLA-A2/HBsAg(335-343) tetramers, secreted IFN-gamma, and lysed target cells presenting a hepatitis B surface antigen (HBsAg) CTL epitope. Enumeration of HBsAg-specific T cells producing cytokine indicated preferential induction of a Type 1 T helper cell response. These results provide the first demonstration of a DNA vaccine inducing protective antibody titers and both humoral and cell-mediated immune responses in humans.

Tolerability and immune responses in humans to a PowderJect DNA vaccine for hepatitis B.

We are developing a DNA vaccine toward hepatitis-B virus (HBV) using PowderJect's proprietary needle-free technology to deliver DNA-coated gold particles directly into cells of the skin. Preclinical studies in animals showed that (i) microgram doses of the DNA vaccine were sufficient to immunize pigs and non-human primates to antibody levels comparable to those obtained with a commercial recombinant subunit vaccine; (ii) the DNA vaccine was effective in mouse strains that respond poorly to protein subunit vaccines; (iii) the vaccine induces robust cytotoxic T-cell responses, and (iv) the vaccine is non-toxic and well tolerated. Based on these findings, this DNA vaccine was evaluated for safety, tolerability, and the induction of immune responses in phase 1 clinical studies in healthy, hepatitis-naïve human volunteers. Preliminary results indicate that the vaccine is safe and well tolerated, and elicits both humoral and cellular immune responses in man.

Preclinical development of human granulocyte-macrophage colony-stimulating factor-transfected melanoma cell vaccine using established canine cell lines and normal dogs.

Tumor vaccines and gene therapy have received significant attention as means of increasing cellular and humoral immune responses to cancer. We conducted a pilot study of seven research dogs to determine whether intradermal injection of canine tumor cells transfected via the Accell particle-mediated gene transfer device with the cDNA for human granulocyte-macrophage colony-stimulating factor (hGM-CSF) would generate biologically relevant levels of protein and result in demonstrable histological changes at sites of vaccination. Tumor cell vaccines of 10(7) irradiated canine melanoma cells were nontoxic, safe, and well tolerated. No significant alterations in blood chemistry values or hematological profiles were detected. A histological review of control vaccine sites revealed inflammatory responses predominated by eosinophils, whereas vaccine sites with hGM-CSF-transfected tumor cells had an influx of neutrophils and macrophages. Enzyme-linked immunosorbent assays of skin biopsies from vaccine sites had local hGM-CSF production (8.68-16.82 ng/site of injection) at 24 hours after injection and detectable levels (0.014-0.081 ng/site) for < or =2 weeks following vaccination. Flow cytometric analysis of hGM-CSF-transfected cells demonstrated < or =25% transfection efficiency, and hGM-CSF levels obtained during time-course assays demonstrated biologically relevant levels for both irradiated and nonirradiated samples. These data demonstrate the in vivo biological activity of irradiated hGM-CSF-transfected canine tumor cells and help provide evidence for a valid translational research model of spontaneous tumors.

Probability analysis of variational crystallization and its application to gp120, the exterior envelope glycoprotein of type 1 human immunodeficiency virus (HIV-1).

The extensive glycosylation and conformational mobility of gp120, the envelope glycoprotein of type 1 human immunodeficiency virus (HIV-1), pose formidable barriers for crystallization. To surmount these difficulties, we used probability analysis to determine the most effective crystallization approach and derive equations which show that a strategy, which we term variational crystallization, substantially enhances the overall probability of crystallization for gp120. Variational crystallization focuses on protein modification as opposed to crystallization screening. Multiple variants of gp120 were analyzed with an iterative cycle involving a limited set of crystallization conditions and biochemical feedback on protease sensitivity, glycosylation status, and monoclonal antibody binding. Sources of likely conformational heterogeneity such as N-linked carbohydrates, flexible or mobile N and C termini, and variable internal loops were reduced or eliminated, and ligands such as CD4 and antigen-binding fragments (Fabs) of monoclonal antibodies were used to restrict conformational mobility as well as to alter the crystallization surface. Through successive cycles of manipulation involving 18 different variants, we succeeded in growing six different types of gp120 crystals. One of these, a ternary complex composed of gp120, its receptor CD4, and the Fab of the human neutralizing monoclonal antibody 17b, diffracts to a minimum Bragg spacing of at least 2.2 A and is suitable for structural analysis.