Cell Culture Modeling to Test Therapies Against Hyperglycemia-Mediated Oxidative Stress and Injury

The concept that oxidative stress is a key mediator of nerve injury in diabetes has led us to design therapies that target oxidative stress mechanisms. Using an in vitro model of glucose-treated dorsal root ganglion (DRG) neurons in culture, we can examine both free radical generation, using fluorimetric probes for reactive oxygen species, and cell death via the TUNEL assay. The cell culture system is scaled down to a 96-well plate format, and so is well suited to high-throughput screening. In the present study, we test the ability of three drugs, nicotinamide, allopurinol, and α-lipoic acid, alone and in combination to prevent DRG neuron oxidative stress and cell death. This combination of drugs is currently in clinical trial in type 1 diabetic patients. We demonstrate independent effects on oxidative stress and neuronal survival for the three drugs, and neuronal protection using the three drugs in combination. The data strengthen the rationale for the current clinical trial. In addition, we describe an effective tool for rapid preclinical testing of novel therapies against diabetic neuropathy. Antioxid. Redox Signal. 7, 1494–1506.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/63115/1/ars.2005.7.1494.pd

Monitoring stormwater contaminants in the Puget Sound nearshore: an active biomonitoring tool using transplanted mussels (Mytilus trossulus)

Stormwater delivers a diverse range of contaminants to receiving waters including Puget Sound. Monitoring stormwater pollutants and their effects on biota is critical to informing best management practices aimed at recovering Puget Sound health. In the winter of 2012/13, the Washington Department of Fish and Wildlife’s Toxics-focused Biological Observation System (TBiOS) team conducted a pilot study using transplanted mussels to characterize the extent and magnitude of contamination in nearshore biota of Puget Sound. Mussels are now a key TBiOS indicator organism for tracking contaminants in the nearshore, and the Stormwater Action Monitoring (SAM) program has adopted mussels for nearshore stormwater monitoring as well. SAM now serves as the primary funder of nearshore mussel monitoring in Puget Sound and the first two SAM mussel monitoring surveys were conducted during the winters of 2015/16 and 2017/18, with future surveys planned on a biennial basis. These mussel surveys utilized native bay mussels (Mytilus trossulus) from a local aquaculture source that were transplanted into anti-predator cages to locations along the Puget Sound shoreline. Monitoring sites covered a broad range of upland land-use types, from rural to highly urban, and concentrations of organic contaminants and metals were measured in the mussels after a two to three-month winter deployment period. Data from the first two years of mussel surveys (2012/13, 2015/16) indicates polycyclic aromatic hydrocarbons (PAHs) and polychlorinated biphenyls (PCBs) were the most abundant organic contaminants of those tested in the nearshore. Concentrations of both contaminants were significantly higher in the most urbanized areas and were positively correlated with impervious surface in upland watersheds adjacent to the nearshore. Patterns of PAHs (i.e. PAH fingerprints) in mussels from different locations demonstrate how mussels might be useful as indicators of sources for this particular class of stormwater contaminants in Puget Sound

Assessing the threat of toxic contaminants to early marine survival of Chinook salmon in the Salish Sea

Human development of the Salish Sea has resulted in loss and modification of salmonid habitats, including reduced habitat quality due to contaminant inputs, particularly in the lower reaches of rivers and estuaries of the central Puget Sound. Chemical contaminants released into the Salish Sea from anthropogenic sources can reduce the health and productivity of salmon. Juvenile salmon are exposed to contaminants in freshwater, estuarine, and marine habitats but they are particularly vulnernable as they transition from fresh to saltwater because this life history stage is especially sensitive to stressors that may reduce their early marine survival. Reduced growth and disease resistance have been demonstrated for juvenile Chinook salmon exposed to environmentally relevant contaminant levels; however, synoptic, Puget Sound-wide surveys to assess the extent and magnitude of contaminant exposure are lacking. In this study we measured exposure of juvenile Chinook salmon to chemicals of concern that enter Puget Sound via stormwater, wastewater treatment facilities, atmospheric deposition to marine waters, and groundwater. During the spring and summer of 2013, outmigrating fish were sampled from the river mouthes and two adjacent marine shorelines at each of five Puget Sound river-estuary systems: Skagit, Snohomish, Green/Duwamish, Puyallup/Hylebos, and Nisqually. We (1) report the extent and magnitude of exposure, (2) compare exposure in outmigrants across five major river-estuary systems, and (3) evaluate potential effects on marine survival. Results will be used to establish a time series of contaminant conditions in juvenile Chinook salmon to measure the effectiveness of current toxics reductions strategies and actions, inform future pollution reduction efforts, and enhanced recovery of Chinook salmon

Identifying factors likely to influence compliance with diagnostic imaging guideline recommendations for spine disorders among chiropractors in North America: a focus group study using the Theoretical Domains Framework

Background: The Theoretical Domains Framework (TDF) was developed to investigate determinants of specific clinical behaviors and inform the design of interventions to change professional behavior. This framework was used to explore the beliefs of chiropractors in an American Provider Network and two Canadian provinces about their adherence to evidence-based recommendations for spine radiography for uncomplicated back pain. The primary objective of the study was to identify chiropractors’ beliefs about managing uncomplicated back pain without xrays and to explore barriers and facilitators to implementing evidence-based recommendations on lumbar spine xrays. A secondary objective was to compare chiropractors in the United States and Canada on their beliefs regarding the use of spine x-rays. Methods: Six focus groups exploring beliefs about managing back pain without x-rays were conducted with a purposive sample. The interview guide was based upon the TDF. Focus groups were digitally recorded, transcribed verbatim, and analyzed by two independent assessors using thematic content analysis based on the TDF. Results: Five domains were identified as likely relevant. Key beliefs within these domains included the following: conflicting comments about the potential consequences of not ordering x-rays (risk of missing a pathology, avoiding adverse treatment effects, risks of litigation, determining the treatment plan, and using x-ray-driven techniques contrasted with perceived benefits of minimizing patient radiation exposure and reducing costs; beliefs about consequences); beliefs regarding professional autonomy, professional credibility, lack of standardization, and agreement with guidelines widely varied (social/professional role & identity); the influence of formal training, colleagues, and patients also appeared to be important factors (social influences); conflicting comments regarding levels of confidence and comfort in managing patients without x-rays (belief about capabilities); and guideline awareness and agreements (knowledge). Conclusions: Chiropractors’ use of diagnostic imaging appears to be influenced by a number of factors. Five key domains may be important considering the presence of conflicting beliefs, evidence of strong beliefs likely to impact the behavior of interest, and high frequency of beliefs. The results will inform the development of a theorybased survey to help identify potential targets for behavioral-change strategies

Input of PBDE exposure in juvenile Chinook salmon along their out-migrant pathway through the Snohomish River, WA

Polybrominated diphenyl ether (PBDE) flame retardant inputs to Puget Sound may be impairing the health of juvenile Chinook salmon and reducing their early marine survival in the Salish Sea, possibly contributing to their decline and limiting their recovery. Previous studies have shown Chinook salmon outmigrating from the Snohomish River accumulate PBDEs at concentrations high enough to alter their immune response, increasing their susceptibility to naturally occurring diseases; however, the source of PBDEs is unknown. Our study objective was to determine where in the Snohomish River system migrating Chinook salmon are exposed to and accumulate PBDEs, and to assess the source so that corrective actions can be implemented. Levels of PBDEs and other persistent organic pollutants were measured in salmon from the upstream tributaries of the Snoqualmie and Skykomish regions, representing the cumulative exposure from all sources prior to entering the Snohomish River, and were compared to those in salmon from subsequent downstream regions of the mainstem to assess where salmon are exposed and the exposure source. Additionally, contaminant levels were measured in salmon sampled from distributary channels of the lower delta to evaluate the extent of PBDE exposure in the outmigrating Snohomish River population. Analyses of the contaminant and body burden data reveal that juvenile Chinook salmon are primarily exposed to and accumulate PBDEs at two sites within the lower delta of the Snohomish River, both located in the immediate vicinity of a wastewater treatment plant outfall. Identification of the region within the Snohomish watershed where salmon are most exposed to PBDEs, as well as the source, allows environmental managers to establish corrective actions to control PBDE inputs. Ultimately, reductions in PBDE exposure should improve the health of Chinook salmon and enhance their marine survival

Susceptibility of optimal train schedules to stochastic disturbances of process times

This work focuses on the stochastic evaluation of train schedules computed by a microscopic scheduler of railway operations based on deterministic information. The research question is to assess the degree of sensitivity of various rescheduling algorithms to variations in process times (running and dwell times). In fact, the objective of railway traffic management is to reduce delay propagation and to increase disturbance robustness of train schedules at a network scale. We present a quantitative study of traffic disturbances and their effects on the schedules computed by simple and advanced rescheduling algorithms. Computational results are based on a complex and densely occupied Dutch railway area; train delays are computed based on accepted statistical distributions, and dwell and running times of trains are subject to additional stochastic variations. From the results obtained on a real case study, an advanced branch and bound algorithm, on average, outperforms a First In First Out scheduling rule both in deterministic and stochastic traffic scenarios. However, the characteristic of the stochastic processes and the way a stochastic instance is handled turn out to have a serious impact on the scheduler performance

Observational and Dynamical Characterization of Main-Belt Comet P/2010 R2 (La Sagra)

We present observations of comet-like main-belt object P/2010 R2 (La Sagra) obtained by Pan-STARRS 1 and the Faulkes Telescope-North on Haleakala in Hawaii, the University of Hawaii 2.2 m, Gemini-North, and Keck I telescopes on Mauna Kea, the Danish 1.54 m telescope at La Silla, and the Isaac Newton Telescope on La Palma. An antisolar dust tail is observed from August 2010 through February 2011, while a dust trail aligned with the object's orbit plane is also observed from December 2010 through August 2011. Assuming typical phase darkening behavior, P/La Sagra is seen to increase in brightness by >1 mag between August 2010 and December 2010, suggesting that dust production is ongoing over this period. These results strongly suggest that the observed activity is cometary in nature (i.e., driven by the sublimation of volatile material), and that P/La Sagra is therefore the most recent main-belt comet to be discovered. We find an approximate absolute magnitude for the nucleus of H_R=17.9+/-0.2 mag, corresponding to a nucleus radius of ~0.7 km, assuming an albedo of p=0.05. Using optical spectroscopy, we find no evidence of sublimation products (i.e., gas emission), finding an upper limit CN production rate of Q_CN<6x10^23 mol/s, from which we infer an H2O production rate of Q_H2O<10^26 mol/s. Numerical simulations indicate that P/La Sagra is dynamically stable for >100 Myr, suggesting that it is likely native to its current location and that its composition is likely representative of other objects in the same region of the main belt, though the relatively close proximity of the 13:6 mean-motion resonance with Jupiter and the (3,-2,-1) three-body mean-motion resonance with Jupiter and Saturn mean that dynamical instability on larger timescales cannot be ruled out.Comment: 23 pages, 13 figures, accepted for publication in A

Effects of polycyclic aromatic hydrocarbons (PAHs) on Pacific herring (Clupea pallasii) embryos exposed to creosote-treated pilings related to a piling removal project in Quilcene Bay, Washington

Fish embryos spawned in Puget Sound nearshore marine habitats face a risk of exposure to a wide variety of toxic chemical pollutants during their incubation. Of particular concern are polycyclic aromatic hydrocarbons (PAHs), chemicals originating from oil spills, combusted fossil fuels, and creosote-treated pilings (CTPs). Removal of CTPs and prohibiting their use in marine waters are two recovery practices aimed at reducing PAHs and other creosote-related chemicals in marine waters. We used manually spawned and field-deployed Pacific herring embryos as a sensitive indicator of PAH exposure from CTPs, to test the efficacy of a CTP removal project in Quilcene Bay Washington. Embryos were deployed near CTPs in a 100-year-old derelict CTP field (1) before the CTPs were removed, (2) just after the removal process, to evaluate whether PAHs were released during removal, and (3) one year later, to evaluate whether PAHs lingered after CTP removal. Embryos incubated in the undisturbed CTP field prior to CTP removal exhibited PAH body burdens approximately five times higher than at reference areas, though total PAHs in the CTP-field embryos were below health effects thresholds. The CTP removal project was not fully completed during this study; CTP debris remained in the piling field and many CTPs were cut at the seafloor, resulting in freshly exposed CTP surfaces after the removal project ended. PAH concentrations in embryos sampled during and after CTP removal were 25x to 83x higher than reference embryos, and many exceeded health effects thresholds. PAH concentrations in embryos after CTP removal correlated with distance from former CTP locations. In addition, expression of cyp1a, a gene involved in PAH-detoxification, was correlated with PAH body burden. These results link embryo health with toxic contaminants associated with CTPs and illustrate the importance of fastidious adherence to appropriate CTP removal protocols to avoid contaminant risks to biota

Systemic Steroid Exposure Is Associated with Differential Methylation in Chronic Obstructive Pulmonary Disease

Rationale: Systemic glucocorticoids are used therapeutically to treat a variety of medical conditions. Epigenetic processes such as DNA methylation may reflect exposure to glucocorticoids and may be involved in mediating the responses and side effects associated with these medications