A comparison of Lowry,Bradford and Smith protein assays using different protein standards and protein isolated from the marine diatom Thalassiosira pseudonana

Two spectrophotometric assays for protein commonly used in marine research (Coomassie stain, Bradford; alkaline copper, Lowry) and a more recent assay which has not been applied in this field (bicinchoninic acid, Smith) were compared for homogenates of the marine diatom Thalassiosira pseudonona using bovine serum albumin (BSA) as a standard. When homogenates were prepared by precipitating protein with trichloroacetic acid (TCA) and redissolving in 1 N NaOH, the protein content estimated by the Lowry and Smith assays agreed closely, but was consistently 20% higher than that indicated by the Bradford assay. To determine if this difference was due to the choice of a protein standard, protein from T. pseudonana was purified and compared to BSA, bovine gamma-globulin (BGG), and casein. The reactivity of the purified protein (expressed as the slope of the absorbance vs protein concentration curve) did not differ between cultures grown at high or low irradiance. For the Smith and Bradford assays the reactivity of BSA was not significantly different from algal protein, but for the Lowry assay, algal protein was significantly higher in reactivity than BSA. BGG was not significantly different in reactivity from algal protein for the Lowry and Smith assays, but BGG gave significantly lower absorbances than algal protein in the Bradford assay. These results suggest that BSA is a suitable standard for algal protein in the Bradford assays, while BGG is preferable for the Lowry assay. Either protein standard could be used for the Smith assay. Differences in purified algal protein reactivity compared to BSA could not account for the differences among the assays, nor could interference by chlorophyll a. Precipitating protein with TCA prior to analyses gave lower protein than direct analyses of homogenates for the Lowry and Smith assays, but no differences were found for the Bradford assay. As a result, the Lowry and Smith assays indicated up to 60% greater protein than the Bradford if TCA precipitation was not performed. This may be due to removal of free amino acids and small peptides which are less reactive in the Bradford assay. The 20% higher protein found in the Lowry or Smith vs Bradford assays may be due to different assay sensitivity to small peptides or other compounds which are precipitated along with proteins by TCA. Although the Smith assay is substantially simpler to perform than the Lowry, there appear to be no quantitative differences in the results. It remains unclear which spectrophotometric assay is most accurate, but the Bradford assay is faster and simpler, and is less likely to be affected by non-protein compounds found in marine phytoplankton.     

Ecological Conservation Through Aesthetic Landscape Planning: A Case Study of the Lower Wisconsin State Riverway

A consequence of expanding residential development into rural areas is the potential alteration of ecological communities. Certain novel land-use policies seek practical solutions by accommodating social needs for housing while conserving biodiversity. This study investigates whether regulations designed to protect the aesthetic characteristics of a river corridor simultaneously mitigate negative effects of development on avian biodiversity, despite the absence of explicit conservation objectives. Using housing data from the US Census (1990 and 2000) and the Audubon Christmas Bird Count (1987–2000), we examined changes in housing density, avian communities, and the relationship between these two variables in a location that has adopted aesthetic landscape planning, the Lower Wisconsin State Riverway. We found that overall species diversity increased in the Riverway, but remained constant in reference areas, although the relative increase in housing density in the two areas did not differ. We also found that omnivore populations decreased in the Riverway and increased in reference sites. On the whole, our study provides preliminary evidence that aesthetic landscape planning, such as employed in the Lower Wisconsin State Riverway, might constitute a politically viable approach to conserve ecological resources.     

Efforts to Reduce Mortality to Hydroelectric Turbine-Passed Fish: Locating and Quantifying Damaging Shear Stresses

Severe fluid forces are believed to be a source of injury and mortality to fish that pass through hydroelectric turbines. A process is described by which laboratory bioassays, computational fluid dynamics models, and field studies can be integrated to evaluate the significance of fluid shear stresses that occur in a turbine. Areas containing potentially lethal shear stresses were identified near the stay vanes and wicket gates, runner, and in the draft tube of a large Kaplan turbine. However, under typical operating conditions, computational models estimated that these dangerous areas comprise less than 2% of the flow path through the modeled turbine. The predicted volumes of the damaging shear stress zones did not correlate well with observed fish mortality at a field installation of this turbine, which ranged from less than 1% to nearly 12%. Possible reasons for the poor correlation are discussed. Computational modeling is necessary to develop an understanding of the role of particular fish injury mechanisms, to compare their effects with those of other sources of injury, and to minimize the trial and error previously needed to mitigate those effects. The process we describe is being used to modify the design of hydroelectric turbines to improve fish passage survival.     

An ultra-clean firn core from the Devon Island Ice Cap, Nunavut, Canada, retrieved using a titanium drill specially designed for trace element studies

An electromechanical drill with titanium barrels was used to recover a 63.7 m long firn core from Devon Island Ice Cap, Nunavut, Canada, representing 155 years of precipitation. The core was processed and analysed at the Geological Survey of Canada by following strict clean procedures for measurements of Pb and Cd at concentrations at or below the pg g(-1) level. This paper describes the effectiveness of the titanium drill with respect to contamination during ice core retrieval and evaluates sample-processing procedures in laboratories. The results demonstrate that: (1) ice cores retrieved with this titanium drill are of excellent quality with metal contamination one to four orders of magnitude less than those retrieved with conventional drills; (2) the core cleaning and sampling protocols used were effective, contamination-free, and adequate for analysis of the metals (Pb and Cd) at low pg g(-1) levels; and (3) results from 489 firn core samples analysed in this study are comparable with published data from other sites in the Arctic, Greenland and the Antarctic.     

The assimilation of contaminants from suspended sediment and algae by the zebra mussel, Dreissena polymorpha

Since their invasion into the Great Lakes, zebra mussels, Dreissena polymorpha, have increased the water clarity in Lake St. Clair and Lake Erie due to their extensive particle filtration. Because these particles contain sorbed contaminants, the potential for contaminant accumulation from both suspended sediment and algae were examined. Sediment or algae were dosed with selected radiolabeled polycyclic aromatic hydrocarbon congeners and/or hexachlorobiphenyl (HCBP). Assimilation efficiencies were measured and depended on food quality. Zebra mussels, 17 ± 2 mm long, assimilated 58.3 ± 13.5% of the pyrene and 44.7 ± 5.8% of the benzo(a)pyrene (BaP) from sediment particles with a particle clearance rate of 493 – 897 ml/g tissue/h. However, assimilation efficiencies were 91.7 ± 3.7% for pyrene, 91.9 ± 1.4% for BaP, 96.6 ± 1.4% for chrysene, and 97.7 ± 0.5% for HCBP from suspended algae. Algal particle clearance rates for the mussels ranged from 47 – 143 ml/g tissue/h. Thus, zebra mussels efficiently accumulated non-polar contaminants sorbed to algae, while a smaller fraction of the sediment-associated contaminant was bioavailable. Furthermore, the contaminants sorbed onto suspended sediment particles were quickly removed from the water and deposited as pseudofeces. The pseudofeces production was positively correlated with filtration rate and suspended particle concentrations.     

Advanced driver assistance systems for teen drivers: A national survey of teen and parent perceptions

Abstract

Objective: Recently developed advanced driver assistance systems (ADAS) have the potential to compensate for teen driving errors and reduce overall crash risk. To date, very limited research has been conducted on the suitability of ADAS for teen drivers—the population most likely to benefit from such systems. The opportunity for ADAS to reduce the frequency and severity of crashes involving teen drivers is hindered when there is a lack of trust, acceptance, and use of those technologies. Therefore, there is a need to study teen and parent perceptions of ADAS to help identify and overcome any potential barriers to ADAS use.

Methods: A U.S. national survey was developed based on themes from previously conducted teen and parent ADAS focus groups. Survey topics included trust in ADAS, effect of ADAS on teen driver safety and driving behavior, effect of ADAS on skill development, data privacy, and cybersecurity. Responses included 5-point Likert scales and open-ended questions. The survey was managed through an online respondent panel by ResearchNow. Eligibility criteria included licensed teens (16–19 years) and parents of licensed teens. Teen and parent responses were compared using chi-square statistics in SAS 9.4.

Results: Two thousand and three (teens?=?1,000; parents?=?1,003) respondents qualified for and completed the survey between September 1 and September 20, 2017. Overall, teens (72%) and parents (61%) felt that ADAS would have a positive impact on transportation. However, teens were more likely to exhibit a positive outlook on ADAS, whereas parents were more likely to have a negative outlook (P?<?.01). Teens felt that ADAS would be useful during bad weather or drowsy driving but were less concerned than parents about ADAS intervention during their own risky driving (P?<?.01). The majority of teens (65%) and parents (71%) agreed that teens should learn to drive on vehicles without ADAS, with parents being more likely to agree than teens (P?<?.01). Parents (55%) were more likely than teens (47%) to be concerned about insurance companies keeping track of teen driving data (P?<?.01). Most respondents exhibited some concern of ADAS being susceptible to hacking (57%).

Conclusions: This study represents the first effort to quantify ADAS perceptions among teen drivers and their parents at the U.S. national level. These data highlight potential barriers to ADAS use among teen drivers, including a relative disinterest among teens for ADAS intervention during risky driving as well as concerns among both teens and parents that ADAS will inhibit skill development. These survey findings will help inform educational programs to accelerate fleet turnover and provide the foundation for ADAS optimization and evaluation studies among sociodemographic groups.     

A Decision Support Framework for Science-Based, Multi-Stakeholder Deliberation: A Coral Reef Example

We present a decision support framework for science-based assessment and multi-stakeholder deliberation. The framework consists of two parts: a DPSIR (Drivers–Pressures–States–Impacts–Responses) analysis to identify the important causal relationships among anthropogenic environmental stressors, processes, and outcomes; and a Decision Landscape analysis to depict the legal, social, and institutional dimensions of environmental decisions. The Decision Landscape incorporates interactions among government agencies, regulated businesses, non-government organizations, and other stakeholders. It also identifies where scientific information regarding environmental processes is collected and transmitted to improve knowledge about elements of the DPSIR and to improve the scientific basis for decisions. Our application of the decision support framework to coral reef protection and restoration in the Florida Keys focusing on anthropogenic stressors, such as wastewater, proved to be successful and offered several insights. Using information from a management plan, it was possible to capture the current state of the science with a DPSIR analysis as well as important decision options, decision makers and applicable laws with a the Decision Landscape analysis. A structured elicitation of values and beliefs conducted at a coral reef management workshop held in Key West, Florida provided a diversity of opinion and also indicated a prioritization of several environmental stressors affecting coral reef health. The integrated DPSIR/Decision landscape framework for the Florida Keys developed based on the elicited opinion and the DPSIR analysis can be used to inform management decisions, to reveal the role that further scientific information and research might play to populate the framework, and to facilitate better-informed agreement among participants.     

The sensitive and selective determination of aluminium by spectrofluorimetric detection after complexation with N-o-vanillidine-2-amino-p-cresol

A simple, easy to use and selective spectrofluorimetric method for the determination of trace levels of aluminium has been developed. A new Schiff base, N-o-vanillidine-2-amino-p-cresol (OVAC), has been synthesized and its fluorescence activity with aluminium investigated. Based on this chelation reaction, a spectrofluorimetric method has been developed for the determination of aluminium in samples buffered at pH 4.0 using acetic acid-sodium acetate. The chelation reaction between Al(iii) and N-o-vanillidine-2-amino-p-cresol was very fast, requiring only 20 min at room temperature to complex completely. The excitation and emission wavelengths were 423.0 and 553.0 nm, respectively, at which the OVAC-Al complex gave the maximum fluorescence intensity at pH 4.0 in a 50% ethanol-50% water medium. The interference from fluoride ions was minimized by the addition of Be(2+). Other ions were found not to interfere at the concentrations likely to be found in natural waters. Under these conditions, the calibration plot was linear up to 1000 microg L(-1) (r = 0.999). The limit of detection (3sigma) for the determination of Al(iii) was 0.19 microg L(-1) and the precision for multiple determinations of 3 ng mL(-1) Al(iii) prepared in ultra-pure water was found to be 0.29% (n = 16). The Schiff base ligand could be used to determine ultra-trace aluminium from natural waters. Analysis of environmental certified reference materials showed good agreement with the certified values. The procedure was found to be equally applicable to both fresh water and saline solutions (including sea water) using either normal external calibration or the standard additions method.     

Effects of the Motor Vehicle Control Program On Hydrocarbon Concentrations in The Central Los Angeles Atmosphere

The body of information presented in this paper is directed to those individuals concerned with the effect of the motor vehicle control program on ambient hydrocarbon concentrations in a metropolitan area during peak traffic hours. The data used in this study are from gas chromatographic analyses of air samples taken in 1963-65, before implementation of the exhaust control program for new motor vehicles, and in 1971 and 1973, after several years’ application of this program. A brief history of the motor vehicle program in California together with emission standards for hydrocarbons are discussed and certain automobile-related hydrocarbons are identified and characterized. Frequency distributions were constructed for total hydrocarbons, non-methane hydrocarbons, methane, acetylene, and isopentane for 1963-65, 1971, and 1973 and the standard deviation (1 sigma) concentration limits are discussed for each of the subgroups mentioned above. The average concentration and relative percent comparisons for methane, ethane, n-butane, isopentane, C₃+ paraffin, eth-ene, propene, C₄ +olefin, acetylene, benzene, and toluene are shown and discussed in detail. These studies: 1) show hydrocarbon emissions from motor vehicles were reduced 47.6% during this time period; 2) indicate selective reduction of compounds; and 3) reveal a 51.8% reduction in the 1973 ambient total hydrocarbon concentration since 1963-65.