Integral assessment of estrogenic potentials in sediment-associated samples

Background, aim, and scope  The enzyme-linked receptor assay (ELRA) detects estrogenic and anti-estrogenic effects at the molecular level of receptor binding and is a useful tool for the integrative assessment of ecotoxicological potentials caused by hormonally active agents (HAA) and endocrine disrupting compounds (EDC). The main advantage of the ELRA is its high sample throughput and its robustness against cytotoxicity and microbial contamination. After a methodological adaptation to salinity of the ELRA, according to the first part of this study, which increased its salinity tolerance and sensitivity for 17-β-estradiol, the optimised ELRA was used to investigate 13 native sediments characterised by different levels of salinity and chemical contamination. The applicability of the ELRA for routine analysis in environmental assessment was evaluated. Salinity is often a critical factor for bioassays in ecotoxicological sediment assessment. Therefore, salinity of the samples was additionally adjusted to different levels to characterise its influence on elution and binding processes of receptor-binding substances. Materials and methods  The ELRA was carried out with the human estrogen receptor α (ER) in a 96-well microplate format using the experimental setup known from the competitive immunoassay based on ligand–protein interaction. It is an important improvement that a physiologically relevant receptor was used as a linking protein instead of an antibody. The microplates were coated with a 17-β-estradiol-BSA conjugate, and dilution series of estradiol and of native sediment samples were added and incubated with the ER. After a washing step, a biotinylated mouse anti-ER antibody was added to each well. Receptor binding to estradiol, agonistic and antagonistic receptor binding, were determined by a streptavidin-POD-biotin complex with subsequent measurement of the peroxidase activity at the wavelength of 450 nm using a commercial ELISA multiplate reader. The sediment elutriates and pore water samples of sediments were tested in a dilution series to evaluate at which dilution step the receptor-binding potential ends. In the elution process (see Section 2.1 to 2.2), a method was developed to adjust the salinity to the levels of the reference testings, which offers an appropriate option to adjust the salinity in both directions. Statistical evaluation was made with a combination of the Mann–Whitney U test and the pT-method. Results  This part of the study characterised the environmental factor ‘salinity’ for prospective applications of the ELRA. Using reference substances such as 17-β-estradiol, the ELRA showed sigmoid concentration-effect relations over a broad range from 0.05 μg/l to 100 μg/l under physiological conditions. After methodological optimisation, both sensitivity and tolerance of the assay against salinity could be significantly raised, and the ELRA became applicable under salinity conditions up to concentrations of 20.5‰. The mean relative inter-test error (n = 3) was around 11% with reference substances and below 5% for single sediments elutriates in three replicates each. For sediment testings, the pore water and different salinity-adjusted elutriates of 13 sediments were used. A clear differentiation of the receptor-binding potential could be reached by application of the pT-method. Thereby, pT-values from one to six could be assigned to the sediments, and the deviation caused by the different salinity conditions was one pT-value. The mean standard deviation in the salinity adaptation procedure of the elutriates was below 5%. Discussion  Although the ELRA has already been used for assessments of wastewater, sludge and soil, its applicability for samples to different salinity levels has not been investigated so far. Even if the ELRA is not as sensitive as the E-screen or the YES-assay, with regard to reference substances like 17-β-estradiol, it is a very useful tool for pre-screening, because it is able to integrate both estrogenic as well as anti-estrogenic receptor-binding effects. According to the results of sediment testing, and given the integrative power to detect different directions of effects, the ELRA shows sufficient sensitivity and salinity tolerance to discriminate receptor-binding potentials in environmental samples. Conclusions  The optimised ELRA assay is a fast, cost-effective, reliable and highly reproducible tool that can be used for high-throughput screening in a microplate format in detecting both estrogenic and anti-estrogenic effects. Additionally, the ELRA is robust against microbial contaminations, and is not susceptible towards cytotoxic interferences like the common cell-culture methods. The general applicability and sufficient sensitivity of the ELRA was shown in freshwater environments. Marine and brackish samples can be measured up to salinity levels of 20.5‰. Recommendations and perspectives  In view of the proven sensitivity, functionality and the fastness of the ELRA, it is recommendable to standardise the test method. At the moment, no adequate in vitro test procedure exists which is standardised to DIN or ISO levels. The E-screen and the yeast estrogen/androgen screens (YES/YAS) sometimes underlie strong cytotoxic effects, as reported in the first part of this study. Further development of an ELRA assay using human androgen receptors appears to be very promising to gain information about androgenic and anti-androgenic effects, too. This would offer a possibility to use the ELRA as a fast and reliable pre-screening tool for the detection of endocrine potentials, thus minimising time and cost-expensive animal experiments.     

Revised estimates of construction activity and emissions: Effects on ozone and elemental carbon concentrations in southern California

Emissions from diesel-powered construction equipment are an important source of nitrogen oxides (NO_x) and particulate matter (PM). A new emission inventory for construction equipment emissions is developed based on surveys of diesel fuel use; the revised inventory is compared to current emission inventories. California's OFFROAD model estimates are 4.5 and 3.1 times greater, for NO_x and PM respectively, than the fuel-based estimates developed here. The most relevant uncertainties are the overall amount of construction activity/diesel fuel use, exhaust emission factors for PM and NO_x, and the spatial allocation of emissions to county level and finer spatial scales. Construction permit data were used in this study to estimate spatial distributions of emissions; the resulting distribution is well correlated with population growth. An air quality model was used to assess the impacts of revised emission estimates. Increases of up to 15 ppb in predicted peak ozone concentrations were found in southern California. Elemental carbon and fine particle mass concentrations were in better agreement with observations using revised emission estimates, whereas negative bias in predictions of ambient NO_x concentrations increased.     

Composting clam processing wastes in a laboratory- and pilot-scale in-vessel system

Waste materials from the clam processing industry (offal, shells) have several special characteristics such as a high salinity level, a high nitrogen content, and a low C/N ratio. The traditional disposal of clam waste through landfilling is facing the challenges of limited land available, increasing tipping fees, and strict environmental and regulatory scrutiny. The aim of this work is to investigate the performance of in-vessel composting as an alternative for landfill application of these materials. Experiments were performed in both laboratory-scale (5L) and pilot-scale (120L) reactors, with woodchips as the bulking agent. In the laboratory-scale composting test, the clam waste and woodchips were mixed in ratios from 1:0.5 to 1:3 (w/w, wet weight). The high ratios resulted in a better temperature performance, a higher electrical conductivity, and a higher ash content than the low-ratio composting. The C/N ratio of the composts was in the range of 9:1-18:1. In the pilot-scale composting test, a 1:1 ratio of clam waste to woodchips was used. The temperature profile during the composting process met the US Environmental Protection Agency sanitary requirement. The final cured compost had a C/N ratio of 14.6, with an ash content of 167.0+/-14.1g/kg dry matter. In addition to the major nutrients (carbon, nitrogen, calcium, magnesium, phosphorus, potassium, sulfur, and sodium), the compost also contained trace amounts of zinc, manganese, copper, and boron, indicating that the material can be used as a good resource for plant nutrients.     

Preferred modes of travel among older adults: What factors affect the choice to walk instead of drive?

Introduction

There are many factors that influence older adults' travel choices. This paper explores the associations between mode of travel choice for a short trip and older adults' personal characteristics.

Methods

This study included 406 drivers over the age of 64 who were enrolled in a large integrated health plan in the United States between 1991 and 2001. Bivariate analyses and generalized linear modeling were used to examine associations between choosing to walk or drive and respondents' self-reported general health, physical and functional abilities, and confidence in walking and driving.

Results

Having more confidence in their ability to walk versus drive increased an older adult's likelihood of walking to make a short trip by about 20% (PR = 1.22; 95% CI: 1.06-1.40), and walking for exercise increased the likelihood by about 50% (PR = 1.53; 95% CI = 1.22-1.91). Reporting fair or poor health decreased the likelihood of walking, as did cutting down on the amount of driving due to a physical problem.

Discussion

Factors affecting a person's decision to walk for exercise may not be the same as those that influence their decision to walk as a mode of travel. It is important to understand the barriers to walking for exercise and walking for travel to develop strategies to help older adults meet both their exercise and mobility needs. Impact on Industry: Increasing walking over driving among older adults may require programs that increase confidence in walking and encourage walking for exercise.     

A note on the effectiveness of the house-arrest alternative for motivating DWI offenders to install ignition interlocks

Introduction

The effectiveness of ignition interlocks at reducing drunk driving has been limited by the ability of driving-while-intoxicated (DWI) offenders to avoid court orders to install the devices.

Methods

In a pilot program in New Mexico, four Santa Fe County judges imposed home confinement (via electronic monitoring bracelets) on offenders who claimed to have no car or no intention to drive. Interlock installation rates for Santa Fe County were compared with all other counties in New Mexico over a 2-year program and 2-year post-program period.

Results

During the two program years, 70% of the drivers convicted of DWI in Santa Fe County installed interlocks, compared to only 17% in the other counties, but when the program was terminated, the Santa Fe installation rate fell by 18.8 percentage points.

Summary

Mandating the alternative sanction of house arrest led to the highest reported interlock installation rate for DWI offenders.

Impact on Industry

Impaired driving is a substantial expense to employers, particularly when it bars driving that interferes with employment. Interlocks provide a method of protecting the public while permitting the offender to drive sober. This study was directed at increasing interlock use by DWI offenders.     

Landscape Correlates of Breeding Bird Richness Across the United States Mid-Atlantic Region

Using a new set of landscape indicator data generated by the U.S.EPA, and a comprehensive breeding bird database from the National Breeding Bird Survey, we evaluated associations between breeding bird richness and landscape characteristics across the entire mid-Atlantic region of the United States. We evaluated how these relationships varied among different groupings (guilds) of birds based on functional, structural, and compositional aspects of individual species demographics. Forest edge was by far the most important landscape attribute affecting the richness of the lumped specialist and generalist guilds; specialist species richness was negatively associated with forest edge and generalist richness was positively associated with forest edge. Landscape variables (indicators) explained a greater proportion of specialist species richness than the generalist guild (46% and 31%, respectively). The lower value in generalists may reflect finer-scale distributions of open habitat that go undetected by the Landsat satellite, open habitats created by roads (the areas from which breeding bird data are obtained), and the lumping of a wide variety of species into the generalist category. A further breakdown of species into 16 guilds showed considerable variation in the response of breeding birds to landscape conditions; forest obligate species had the strongest association with landscape indicators measured in this study (55% of the total variation explained) and forest generalists and open ground nesters the lowest (17% of the total variation explained). The variable response of guild species richness to landscape pattern suggests that one must consider species' demographics when assessing the consequences of landscape change on breeding birds.     

Design and evaluation of an inlet conditioner to dry particles for real-time particle sizers

Real-time particle sizers provide rapid information about atmospheric particles, particularly peak exposures, which may be important in the development of adverse health outcomes. However, these instruments are subject to erroneous readings in high-humidity environments when compared with measurements from filter-based, federal reference method (FRM) samplers. Laboratory tests were conducted to evaluate the ability of three inlet conditioners to dry aerosol prior to entering a real-time particle sizer for measuring coarse aerosols (Model 3321 Aerodynamic Particle Sizer, APS) under simulated highly humid conditions. Two 30 day field studies in Birmingham, AL, USA were conducted to compare the response of two APSs operated with and without an inlet conditioner to that measured with FRM samplers. In field studies, the correlation of PM(10-2.5) derived from the APS and that measured with the FRM was substantially stronger with an inlet conditioner applied (r2 ranged from 0.91 to 0.99) than with no conditioner (r2 = 0.61). Laboratory experiments confirmed the ability of the heater and desiccant conditioner to remove particle-borne moisture. In field tests, water was found associated with particles across the sizing range of the APS (0.5 microm to 20 microm) when relative humidity was high in Birmingham. Certain types of inlet conditioners may substantially improve the correlation between particulate mass concentration derived from real-time particle sizers and filter-based samplers in humid conditions.     

Managing scuba divers to meet ecological goals for coral reef conservation

Marine protected areas increasingly are challenged to maintain or increase tourism benefits while adequately protecting resources. Although carrying capacity strategies can be used to cope with use-related impacts, there is little understanding of divers themselves, their management preferences, and how preferences relate to conservation goals. By using a stated preference choice modeling approach, we investigated the choices divers make in selecting diving trips to marine protected areas as defined by use level, access, level of supervision, fees, conservation education, and diving expectations. Logit models showed that divers preferred a more restrictive management scenario over the status quo. Divers favored reductions in the level of site use and increased levels of conservation education. Divers did not favor fees to access protected areas, having less access to the resource, or extensive supervision. Finally, divers were much more willing to accept increasingly restrictive management scenarios when they could expect to see increased marine life.