Sample size and power determination for detecting interactions in mixtures of chemicals

In the analysis of mixtures of drugs/chemicals it is often of interest to test for the presence of interaction. If the hypothesis of no interaction (additivity) is not rejected, then the analyst may reasonably claim additivity if and only if the study is powered to a desired (e.g., biologically meaningful) level. The objective of this article is to address the sample size and power issues related to testing the hypothesis of additivity at specified mixture points. The study of disinfectant by-products (DBPs) found in drinking water, described in earlier literature, is used to illustrate the procedures for estimating power and sample sizes for detecting interactions at specified mixtures. The four trihalomethanes used in the study are bromodichloromethane (BDCM), chlorodibromomethane (CDBM), chloroform (CHCl₃), and bromoform (CHBr₃)     

Bioassay-directed Fractionation of Organic Extracts of Marine Surface Sediments from the North and Baltic Sea - Part II: Results of the biotest battery and development of a biotest index (8 pp)

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Part I: Determination and identification of organic pollutants Part II: Results of the biotest battery and development of a biotest index

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Preamble. This series of two papers presents the results of an interdisciplinary research project (ISIS) dealing with bioassay-directed fractionation of marine sediment extracts. Part I presents the extraction and fractionation procedure as well as the results of chemical analysis, including non-target analysis of sediments. Part II describes the results of the biotest battery in relation to chemicals possibly causing parts of the observed effects. A biotest index is used to compare the toxicities of the samples.

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AUTHORS / AFFILIATIONS Ninja Reineke (3), Werner Wosniok (4), Dirk Danischewski (1), Heinrich Hühnerfuss (3), Angelika Kinder (5), Arne Sierts-Herrmann (5), Norbert Theobald (2), Hans-Heinrich Vahl (6), Michael Vobach (1), Johannes Westendorf (6) and Hans Steinhart (5).

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(1) Federal Research Centre for Fisheries, Institute for Fishery Ecology, Palmaille 9, 22767 Hamburg, Germany (2) Federal Maritime and Hydrographic Agency, Bernhard-Nochtstr. 78, 20359 Hamburg, Germany (3) University of Hamburg, Institute for Organic Chemistry, Martin-Luther-King-Platz 6, 20146 Hamburg, Germany (4) University of Bremen, Institute of Statistics, Bibliothekstr. 1, 28334 Bremen, Germany (5) University of Hamburg, Institute for Food Chemistry, Grindelallee 117, 20146 Hamburg, Germany (6) University of Hamburg, University Hospital Hamburg-Eppendorf, Department for Toxicology, Vogt-Kölln-Str. 30, 22527 Hamburg, Germany (7) Eurofins Wiertz-Eggert-Jörissen, Stenzelring 14b, 21107 Hamburg, Germany

Goal, Scope and Background

The ecological relevance of contaminants in mixtures is difficult to assess, because of possible interactions and due to lacking toxicity data for many substances present in environmental samples. Marine sediment extracts, which contain a mixture of environmental contaminants in low concentrations, were the object of this study. The extracts were investigated with a set of different biotests in order to identify the compound or the substance class responsible for the toxicity. For this goal, a combination of biotests, biotest-directed fractionation and chemical analysis has been applied. Further on, a strategy for the development of a biotest index to describe the toxicity of the fractions without a prior ranking of the test results is proposed. This article (Part II) focuses on the biological results of the approach.

Methods

The toxicological potential of organic extracts of sediments from the North Sea and the Baltic Sea was analyzed in a bioassay-directed fractionation procedure with a set of biotests: luciferase reporter gene assays on hormone receptor and Ah receptor, arabinose resistance test, fish embryo test (Danio rerio), comet assay, acetylcholinesterase inhibition test, heat-shock protein 70 induction, oxidative stress and luminescence inhibition test (Vibrio fischeri). The test results provided the basis for the calculation of a biotest index by factor analysis to compare the toxicity of the samples and fractions.

Results and Discussion

Results of 11 biotests on different fractionation levels of the samples were described and discussed with regard to the occurrence of contaminants and their toxic potentials. Polychlorinated biphenyls, polycyclic aromatic hydrocarbons, quinones, brominated indoles and brominated phenols were in the focus of interest. A biotest index was constructed to compare the toxic responses in the samples and to group the biotest results.

Conclusion

The procedure presented in this study is well suited for bioassay-directed fractionation of marine sediment extracts. However, in relatively low contaminated samples, high enrichment factors and sufficient fractionation is necessary to allow identification of low concentrations of contaminants which is required to link effects and possible causes. In the present case, the relation between substances and effects was difficult to uncover due to relatively low concentrations of pollutants compared to the biogenic matrix and to the remaining complexity of the fractions. The results, with respect to the brominated phenols and indoles in the samples, highlight the successful use of bioassay directed fractionation in the case of high concentrations and high toxicity.

Recommendation and Outlook

In general, it has been shown that a marine risk assessment requires focusing on the input of diffuse sources and taking into account the fact of mixture toxicity. Effects resulting from biogenic substances will make the assessment of the influence of anthropogenic substances even more difficult.     

The waldrapp, Geronticus eremita (Linnaeus, 1758): Historical review, taxonomic history, and present status

The Waldrapp ibis, Geronticus eremita Linnaeus, 1758, is known historically from Ancient Egypt, the Alps in southern Europe, the Near and Middle East, Morocco, and Algeria. This paper summarises what is known about the species from archaeological findings and historical expeditions to these regions, and outlines the taxonomic confusion which characterised this bird for four centuries—the 16th to 19th.

The ancient and recent decline of populations of the Waldrapp is discussed. The birds are at present restricted to about twelve breeding sites, totalling only about 400 individuals, in Turkey and Morocco, with an additional 400-plus specimens in captivity in zoos of the world. A plea is made for the conservation of this endangered species.     

Monitoring of heavy metals in protected forest catchments in Sweden

Long-term integrated catchment monitoring within the Swedish Environmental Monitoring Programme (PMK) aims primarely at the fate and effects of pollutants in mature ecosystems, mainly protected boreal forests. The measurements that were conducted since the early eighties, with some variation, included deposition, throughfall, litterfall and the chemistry of soil water, groundwater and runoff water.Together with, or perhaps partly due to acidification, accumulation and transport of heavy metals are a serious threat to the ecosystem and in the end to the health of human beings. This paper discuss the behaviour of Hg, Pb, Cd, Zn and Cu based on the experiece gained from the studies in the small catchment areas. The importance of present deposition, soil storage and biogeochemical condition for metal mobility to the transport of metals is discussed The storages of Hg and Pb have been built up during a long time span and are very large in comparison with the flux. The present atmospheric deposition have probably little impact on the flux, which rather is governed by the biogeochemical conditions. There is still a small accumulation of Cd, but its great mobility under acidic conditions makes input and output almost in balance. A reduction in deposition will probably have an immediate effect on flux, which also is the case for Zn. Cu generally accumulates in soil. The mobility and hence the flux are certainly regulated by the biogeochemical conditions, but more detailed studies are required to reveal the general mecanisms for Cu transport within the catchment.     

Handling missingness when modeling the force of infection from clustered seroprevalence data

Modeling infectious diseases data is a relatively young research area in which clustering and stratification are key features. It is not unlikely for these data to have missing values. If values are missing completely at random, the analysis on the complete cases is valid. However, in practice this assumption is usually not fulfilled. This article shows the effect of ignoring missing data in modeling the force of infection of the bovine herpesvirus-1 in Belgian cattle and proposes the use of weighted generalized estimating equations with constrained fractional polynomials as a flexible modeling tool.     

Impacts from Deposition on Swedish Forest Ecosystems Identified by Integrated Monitoring

Integrated monitoring of ecosystems (IM) is an international co-operative programme (ICP) to control effects of air pollution and climate change on water, soil and biological systems. It is a part of the Convention on Long-Range Transboundary Air Pollution (CLRTAP) of the United Nations Economic Commission of Europe (UN/ECE). The ICP-IM is undertaken on sites/catchments to investigate acidification, eutrophication and heavy metals with an integrated approach. In Sweden, long-term time series from forest ecosystems, with a long and stable continuity, will reveal trends and changes in processes and enable modelling to be undertaken. Investigations of acidity/alkalinity in relation to mineral and organic acids indicated the importance of atmospheric deposition. Recent results show very high inorganic nitrogen retention (99%), a net loss of sulphur originating mainly from organic horizons, and a high inorganic aluminium content in the illuvial soil horizons which could be detrimental to forests. Forest deficiency could also be caused by an observed ongoing translocation of Zn to deeper soil layers implying a movement towards increased release to surface waters.     

Die Interpretation von K/Ca-Austauschkurven mit Hilfe einer einfachen GAPON-Gleichung

The interpretation of K/Ca exchange isotherms by a simple GAPON equation K/Ca exchange isotherms of various soils can be described by a GAPON equation, where the coefficients being fitted to the measured values by iteration. The resulting values for the “labile potassium” are very similar to those of exchangeable K, whereas only 5–25% of the cec is involved in the short-term K/Ca exchange.