A tool for rapid screening of direct DNA agents using reaction rates and relative interaction potency: towards screening environmental contaminants for hazard

DNA damage represents a potential biomarker for determining the exposure risk to chemicals and may provide early warning data for identifying chemical hazards to human health. Here, we have demonstrated a simple chromatography-based method that can be used to rapidly screen for the presence of chemical hazards as well as to determine parameters relevant to hazard assessment. In this proof-of-principle study, a simple in vitro system was used to determine the interaction of pollutants and probable carcinogens, phenyl glycidyl ether (PGE), tetrachlorohydroquinone (Cl(4)HQ), methylmethane sulfonate (MMS), styrene-7,8-oxide (SO), and benzo[a]pyrene-7,8-dihydrodiol-9,10-epoxide (BPDE), a metabolite of benzo[a]pyrene (B[a]P), with single- and double-stranded DNA probes. Differences in potency and reaction kinetics were studied for chemical and DNA type. A relative interaction potency equivalency (PEQ) of a chemical was determined by ratio of interaction potency of a chemical to BPDE as the reference chemical in the reaction with single- and double-stranded oligodeoxynucleotides. PEQs were found to be BPDE > PGE > SO > MMS > Cl(4)HQ for single-stranded oligodeoxynucleotides while they were found to be BPDE > PGE > Cl(4)HQ > MMS > SO for double-stranded oligodeoxynucleotides. Kinetics evaluation revealed that BPDE reacted with both DNA probes at a significantly faster rate, as compared to the remaining test chemicals. Equilibrium was reached within an hour for BPDE, but required a minimum of 48 h for the remaining chemicals. First-order rate constants were (1.61 ± 0.2) × 10(-3) s(-1) and (3.18 ± 0.4) × 10(-4) s(-1) for reaction of BPDE with double- and single-stranded DNA, respectively. The remaining chemicals possessed rate constants from 2 to 13 × 10(-6) s(-1) with a relative kinetic order for reaction with DNA of BPDE ? MMS > SO > PGE > Cl(4)HQ for ds-DNA and BPDE ? SO ≈ Cl(4)HQ ≈ MMS > PGE for ss-DNA. We further found that the reaction potency, defined by dose-response between chemical pollutants and DNA, depends on the form of DNA present for reaction. Noteworthy, we found that relative PEQ did not follow the same kinetic trends. However, our preliminary findings suggest that reaction kinetics, in combination with relative interaction potency, may be a significant parameter that can be used to evaluate the hazard level of environmental pollutants.     

APPLICATION OF THE BASINS DATABASE AND NPSM MODEL ON A SMALL OHIO WATERSHED1

ABSTRACT: The purpose of this study was to evaluate the Better Assessment Science Integrating Point and Nonpoint Sources (BASINS) watershed management system. BASINS data were used with the NPSM model to predict discharge and sediment concentrations at the outlet of a 103 km² Ohio watershed. It was concluded that the NPSM model should always be calibrated but only a few of the parameters provided with BASINS needed to be calibrated. For a three‐year study period, there was a 2 percent underestimation of discharge using area weighted precipitation values and a 25 percent overestimation using the single station data in BASINS. A comparison of observed and predicted monthly discharge resulted in an r² of 0.86 with area‐weighted precipitation and an r² of 0.74 with the single station data. Calibrating the model to substantially improve sediment predictions was unsuccessful and we concluded that a calibration period of one year was too short. For the three‐year study period, the r² for sediment was 0.36 with a slope of 0.37 and an intercept of 18.8 mg/l. The mean observed and predicted sediment concentrations were 27.1 mg/l and 22.6 mg/l, respectively.     

Spatial spread of an emerging infectious disease: conjunctivitis in House Finches

In this paper we quantify the rate of spread of the newly emerged pathogen Mycoplasma gallisepticum of the House Finch, Carpodacus mexicanus, in its introduced range. We compare and contrast the rapid, yet decelerating, rate of spread of the pathogen with the slower, yet accelerating rate of spread of the introduced host. Comparing the rate of spread of this pathogen to pathogens in terrestrial mammalian hosts, we see that elevation and factors relating to host abundance restrict disease spread, rather than finding any major effects of discrete barriers or anthropogenic movement. We examine the role of seasonality in the rate of spread, finding that the rate and direction of disease spread relates more to seasonality in host movement than to seasonality in disease prevalence. We conclude that asymptomatic carriers are major transmitters of Mycoplasma gallisepticum into novel locations, a finding which may also be true for many other diseases, such as West Nile Virus and avian influenza.     

A field study of the influence of land management and soil properties on runoff and soil loss in central Spain

Since the 1950s afforestation of degraded land has been the principal means of combatting erosion in a seasonally arid area of central Spain. In the 1970s tree planting of steep hillsides and gully sides was preceded by bench terracing. Experimental sites have been established to monitor runoff and soil losses under mature Pinus forest, 12-year-old Pinus forest, and Cistus matorral. The experiment is being conducted at three scales: large gully or small watershed (c. 3.5 ha), runoff erosion plot (10–21.5 m²), and rainfall simulation plot (1 m²). Monitoring began in October 1992. Discharge was recorded continuously, while sediment loss and soil moisture content were measured on a storm basis. The paper presents summary data on runoff and soil erosion for the three scales and comments on relationships between land management, site characteristics, and these losses. We stress the crucial role of vegetation and its interrelationship with soil properties such as structure and aggregate stability. Matorral was effective in combatting water and soil loss, but we question the practice of afforesting seasonally arid, steeply sided areas that have highly erodible soils.     

Detecting river pollution using fluorescence spectrophotometry: case studies from the Ouseburn,NE England

Recent advances in fluorescence spectrophotometry enable the analysis of river dissolved organic matter. We investigate the potential of detecting sewage pollution in a small, urbanised catchment. Downstream sampling highlighted a summer maximum in tryptophan fluorescence intensity during low flow. No correlation is observed between ammonia and tryptophan fluorescence intensity. In contrast, two sewage related point-pollution events had both high tryptophan fluorescence intensity and ammonia, suggesting that the summer tryptophan increase does not original from foul sewage. Sewage inputs to the river were therefore monitored at summer base flow. This demonstrated that > 10% of the rivers' discharge is provided by sewerage inputs and that these inputs could be grouped by their fluorescence and ammonia properties: (1) 'clean' storm waters with low ammonia and tryptophan intensity (2) 'grey' waters with high tryptophan intensity and low ammonia concentration, and (3) 'foul' waters with high tryptophan intensity and ammonia concentration. All three types of sewerage input occurred irrespective of flow conditions,suggesting that sewerage cross connections are occurring.     

Nutrients on Asphalt Parking Surfaces in an Urban Environment

Amounts of readily soluble nutrients on asphalt parking lot surfaceswere measured at four locations in metropolitan Phoenix, Arizona, U.S.A. Using a rainfall simulator, short intense rainfall events were generated to simulate `first flush' runoff. Samples were collected from 0.3 m² sections of asphalt at 8 to 10 sites on each of four parkinglots, during the pre-monsoon season in June-July 1998 and analyzed for dissolved NO₃ ^--N, NH₄ ⁺-N, soluble reactive phosphate (SRP), and dissolved organic carbon (DOC). Runoff concentrations varied considerably for NO₃ ^--N and NH₄ ⁺-N (between 0.1 and 115.8 mg L^-1) and DOC (26.1 to 295.7 mg L^-1), but less so for SRP (0.1 to 1.0 mg L^-1), representing average surface loadings of 191.3, 532.2, and 1.8 mg m^-2 respectively. Compared with similar data collected from undeveloped desert soil surfaces outside the city, loadings of NO₃ ^--N and NH₄ ⁺-N on asphalt surfaces were greater by factors of 91 and 13, respectively. In contrast, SRP loads showed little difference between asphalt and desert surfaces. Nutrient fluxes in runoff from a storm that occurred shortly after the experiments were used to estimate input-output budgets for 3 of the lots under study. Measured outputs of DOC and SRP were similar to those predicted using rainfall and experimentally determined surface loadings, but for NH₄ ⁺-N and particularly for NO₃ ^--N, estimated rainfall inputs and surface runoff were significantly higher than exports in runoff. This suggests that parking lots may be important sites for nutrient accumulation and temporary storage in arid urban catchments.     

Plant growth rates and seed size: a re-evaluation

Small-seeded plant species are often reported to have high relative growth rate or RGR. However, because RGR declines as plants grow larger, small-seeded species could achieve higher RGR simply by virtue of their small size. In contrast, size-standardized growth rate or SGR factors out these size effects. Differences in SGR can thus only be due to differences in morphology, allocation, or physiology. We used nonlinear regression to calculate SGR for comparison with RGR for 10 groups of species spanning a wide range of life forms. We found that RGR was negatively correlated with seed mass in nearly all groups, but the relationship between SGR and seed mass was highly variable. We conclude that small-seeded species only sometimes possess additional adaptations for rapid growth over and above their general size advantage.