The OECD Validation Program of the H295R Steroidogenesis Assay for the Identification of In Vitro Inhibitors and Inducers of Testosterone and Estradiol Production. Phase 2: Inter-Laboratory Pre-Validation Studies (8 pp)

Background, Goals and Scope In response to concerns that have been raised about chemical substances that may alter the function of endocrine systems and result in adverse effects on human health, an OECD initiative was undertaken to develop and validate in vitro and in vivo assays to identify chemicals that may interfere with endocrine systems of vertebrates. Here we report on studies that were conducted to develop and standardize a cell-based screening assay using the H295R cell line to prioritize chemicals that may act on steroidogenic processes in humans and wildlife. These studies are currently ongoing as part of the ‘Special Activity on the Testing and Assessment of Endocrine Disruptors’ within the OECD Test Guidelines Program to review, develop, standardize, and validate a number of in vitro and in vivo toxicological assays for testing and assessment of chemicals concerning their potential to interact with the endocrine system of vertebrates. Study Design Six laboratories from five countries participated in the pre-validation studies. Each laboratory tested the effects of three model chemicals on the production of testosterone (T) and estradiol (E2) using the H295R Steroidogenesis Assay. Chemicals tested were well described inducers or inhibitors of steroidogenic pathways (forskolin, prochloraz and fadrozole). All experiments were conducted in 24 well plates following standard protocols. Six different doses per compound were analyzed in triplicate per plate. A quality control (QC) plate was run in conjunction with the chemical exposure plate to account for inter-assay variation. Each chemical exposure was conducted two or three times. Results All laboratories successfully detected increases and/or decreases in hormone production by H295R cells after exposure to the different model compounds and there was good agreement in the pattern of response for all groups. Forskolin increased both T and E2 while fadrozole and prochloraz decreased production of both hormones. All chemicals affected hormone production in a dose-dependent manner with the exception of fadrozole which caused maximum inhibition of E2 at the two least concentrations tested. Some inter-laboratory differences were noted in the alteration of hormone production measured in chemically exposed cells. However, with the exception of the production of T measured at one laboratory in cells exposed to forskolin, the EC₅₀s calculated were comparable (coefficients of variation 34–49%) for all hormones. Discussion and Perspectives The results indicated that the H295R Steroidogenesis Assay protocol was robust, transferable and reproducible among all laboratories. However, in several instances that were primarily related to one laboratory there were unexplained minor uncertainties related to the inter-laboratory hormone production variation. Based on the findings from this Phase 2 prevalidation study, the H295R Steroidogenesis Assay protocol is currently being refined. The next phase of the OECD validation program will test the refined protocol among the same group of laboratories using an extended set of chemicals (∼30) that will include positive and negative chemical controls as well as a broad spectrum of different potential inducers and inhibitors of steroidogenic pathways. Submission Editor: Dr. Carsten Brühl (bruehl@uni-landau.de)     

Human Culture and Large Carnivore Conservation in North America

We examined and compared human attitudes toward wolves ( Canis lupus ), grizzly bears ( Ursus arctos horribilis ), and mountain lions ( Puma concolor ) in North America, with an emphasis on the Rocky Mountains of the United States and Canada. Primary research, literature review, and secondary data were included in the analysis. Wolves were historically persecuted by Euro-American settlers, but they have been the focus of a significant attitudinal transformation during the latter half of this century. Many now view the wolf in positive and protective ways. Considerable variation remains, however, in contemporary attitudes. Attitudes toward mountain lions have been highly ambivalent, never assuming the prominence or clarity of views on wolves. The behavior and physiology of the species, along with human psychological factors, are important reasons for the difference in human perceptions of wolves and mountain lions. We also examined attitudes toward grizzly bears. Many indigenous cultures revered grizzly bears, although European settlers were interested in exploiting and eliminating them. Today, perceptions of grizzly bears range from positive to negative. Our recommendations include targeting key groups with education programs, building support through the use of spokespeople within the target groups, integrating human and ecological concerns, and designing species-specific education initiatives in some cases.     

Hierarchical modeling of seed variety yields and decision making for future planting plans

Eradicating hunger and malnutrition is a key development goal of the twenty first century. This paper addresses the problem of optimally identifying seed varieties to reliably increase crop yield within a risk-sensitive decision making framework. Specifically, a novel hierarchical machine learning mechanism for predicting crop yield (the yield of different seed varieties of the same crop) is introduced. This prediction mechanism is then integrated with a weather forecasting model and three different approaches for decision making under uncertainty to select seed varieties for planting so as to balance yield maximization and risk. The model was applied to the problem of soybean variety selection given in the 2016 Syngenta Crop Challenge. The prediction model achieved a median absolute error of 235 kg/ha and thus provides good estimates for input into the decision models. The decision models identified the selection of soybean varieties that appropriately balance yield and risk as a function of the farmer’s risk aversion level. More generally, the models can support farmers in decision making about which seed varieties to plant.     

Founder need to belong,tertius iungens orientation and new venture performance

In this study, we examine the role of founders' need to belong and tertius iungens orientation (i.e., the tendency of bringing people together and coordinating among others) in the context of new ventures. We propose and document evidence that a founder's tendency to engage in tertius iungens oriented networking is positively associated with new venture performance. We also reveal that a founder's need to belong is a significant motivational antecedent of the founder's tertius iungens orientation. Further, the need to belong–networking orientation linkage is stronger for founders in new ventures at the early product development stage than those at the late product development stage. We tested our hypotheses using survey data collected with a two‐wave sample of new ventures in China. We discuss the theoretical significance of studying the need to belong and tertius iungens orientation for social networks and entrepreneurship literatures, as well as the practical implications for managing a successful new venture.     

High solids co-digestion of food and landscape waste and the potential for ammonia toxicity

A pilot-scale study was completed to determine the feasibility of high-solids anaerobic digestion (HSAD) of a mixture of food and landscape wastes at a university in central Pennsylvania (USA). HSAD was stable at low loadings (2 g COD/L-day), but developed inhibitory ammonia concentrations at high loadings (15 g COD/L-day). At low loadings, methane yields were 232 L CH₄/kg COD fed and 229 L CH₄/kg VS fed, and at high loadings yields were 211 L CH₄/kg COD fed and 272 L CH₄/kg VS fed. Based on characterization and biodegradability studies, food waste appears to be a good candidate for HSAD at low organic loading rates; however, the development of ammonia inhibition at high loading rates suggests that the C:N ratio is too low for use as a single substrate. The relatively low biodegradability of landscape waste as reported herein made it an unsuitable substrate to increase the C:N ratio. Codigestion of food waste with a substrate high in bioavailable carbon is recommended to increase the C:N ratio sufficiently to allow HSAD at loading rates of 15 g COD/L-day.     

Bioventing of No. 2 Fuel Oil: Effects of Air Flowrate,Temperature, Nutrient Amendment,and Acclimation

A 14‐month pilot‐scale bioventing study, sponsored by the New Hampshire Department of Environmental Services, was conducted by the University of New Hampshire to determine the effects of the time between the contamination event and the onset of bioventing, as well as air flowrate, temperature, and nutrient amendments. Freshly contaminated soil was not readily amenable to bioventing. Bioventing was effective (82–92.5 percent removal) for acclimated soil amended with nutrients at 10 °C and 20 °C for the 275 cm³/min and 140 cm³/min air flowrates, respectively. First order degradation rates after nutrient addition were ?6.11 ± 0.83 (×10^?3)/day and ?6.57 ± 1.71 (×10^?3)/day, respectively. The results indicate that bioventing will be best applied when the contamination has occurred at least two years before the onset of treatment. © 2014 Wiley Periodicals, Inc.     

Trends and applications of multi-criteria decision analysis: use in government agencies

Government agencies are responsible for making complex, high-stake decisions, which require them to balance political, technical, and economic considerations. Pressure from stakeholders and administrative requirements necessitate a traceable and transparent method for decision making. Multi-criteria decision analysis (MCDA) methods are available to decision makers to facilitate systematic treatment of the information and factors necessary to make informed and effective decisions in complex circumstances. A survey of gray and academic literature was conducted to gauge the level of application and awareness of MCDA methods by US government agencies and determine if the tools’ benefits are being realized. Results show an increase in awareness and consideration of MCDA from 2000 to the present, and that agencies are especially considering and using tools to engage with stakeholders. Government agencies would benefit from extending the application of MCDA to strategic planning and congressional engagement, as well as by standardizing MCDA use to better enable inter-agency collaboration and communication.     

Critical Review of Exposure and Effects: Implications for Setting Regulatory Health Criteria for Ingested Copper

Environmental Management - Decades of study indicate that copper oral exposures are typically not a human health concern. Ingesting high levels of soluble copper salts can cause acute...     

Speciated arsenic in air: measurement methodology and risk assessment considerations

Accurate measurement of arsenic (As) in air is critical to providing a more robust understanding of arsenic exposures and associated human health risks. Although there is extensive information available on total arsenic in air, less is known on the relative contribution of each arsenic species. To address this data gap, the authors conducted an in-depth review of available information on speciated arsenic in air. The evaluation included the type of species measured and the relative abundance, as well as an analysis of the limitations of current analytical methods. Despite inherent differences in the procedures, most techniques effectively separated arsenic species in the air samples. Common analytical techniques such as inductively coupled plasma mass spectrometry (ICP-MS) and/or hydride generation (HG)- or quartz furnace (GF)-atomic absorption spectrometry (AAS) were used for arsenic measurement in the extracts, and provided some of the most sensitive detection limits. The current analysis demonstrated that, despite limited comparability among studies due to differences in seasonal factors, study duration, sample collection methods, and analytical methods, research conducted to date is adequate to show that arsenic in air is mainly in the inorganic form. Reported average concentrations of As(III) and As(V) ranged up to 7.4 and 10.4 ng/m3, respectively, with As(V) being more prevalent than As(III) in most studies. Concentrations of the organic methylated arsenic compounds are negligible (in the pg/m3 range). However because of the variability in study methods and measurement methodology, the authors were unable to determine the variation in arsenic composition as a function of source or particulate matter (PM) fraction. In this work, the authors include the implications of arsenic speciation in air on potential exposure and risks. The authors conclude that it is important to synchronize sample collection, preparation, and analytical techniques in order to generate data more useful for arsenic inhalation risk assessment, and a more robust documentation of quality assurance/quality control (QA/QC) protocols is necessary to ensure accuracy, precision, representativeness, and comparability.