Lessons Learned from Read-Across Case Studies for Repeated-Dose Toxicity

A series of case studies designed to further acceptance of read-across predictions, especially for chronic health-related endpoints, have been evaluated with regard to the knowledge and insight they provide. A common aim of these case studies was to examine sources of uncertainty associated with read-across. While uncertainty is related to the quality and quantity of the read across endpoint data, uncertainty also includes a variety of other factors, the foremost of which is uncertainty associated with the justification of similarity and quantity and quality of data for the source chemical(s). This investigation has demonstrated that the assessment of uncertainty associated with a similarity justification includes consideration of the information supporting the scientific arguments and the data associated with the chemical, toxicokinetic and toxicodynamic similarity. Similarity in chemistry is often not enough to justify fully a read-across prediction, thus, for chronic health endpoints, toxicokinetic and/or toxicodynamic similarity is essential. Data from New Approach Methodology(ies) including high throughput screening, in vitro and in chemico assay and in silico tools, may provide critical information needed to strengthen the toxicodynamic similarity rationale. In addition, it was shown that toxicokinetic (i.e., ADME) similarity, especially metabolism, is often the driver of the overall uncertainty

Evolutionary variation in the expression of phenotypically plastic color vision in Caribbean mantis shrimps, genus Neogonodactylus

Author Posting. © The Author(s), 2006. This is the author's version of the work. It is posted here by permission of Springer for personal use, not for redistribution. The definitive version was published in Marine Biology 150 (2006): 213-220, doi:10.1007/s00227-006-0313-5.Many animals have color vision systems that are well suited to their local environments. Changes in color vision can occur over long periods (evolutionary time), or over relatively short periods such as during development. A select few animals, including stomatopod crustaceans, are able to adjust their systems of color vision directly in response to varying environmental stimuli. Recently, it has been shown that juveniles of some stomatopod species that inhabit a range of depths can spectrally tune their color vision to local light conditions through spectral changes in filters contained in specialized photoreceptors. The present study quantifies the potential for spectral tuning in adults of three species of Caribbean Neogonodactylus stomatopods that differ in their depth ranges to assess how ecology and evolutionary history influence the expression of phenotypically plastic color vision in adult stomatopods. After 12 weeks in either a full-spectrum “white” or a narrow-spectrum “blue” light treatment, each of the three species evidenced distinctive tuning abilities with respect to the light environment that could be related to its natural depth range. A molecular phylogeny generated using mitochondrial cytochrome oxidase C subunit 1 (CO-1) was used to determine whether tuning abilities were phylogenetically or ecologically constrained. Although the sister taxa N. wennerae and N. bredini both exhibited spectral tuning, their ecology (i.e. preferred depth range) strongly influenced the expression of the phenotypically plastic color vision trait. Our results indicate that adult stomatopods have evolved the ability to undergo habitat-specific spectral tuning, allowing rapid facultative physiological modification to suit ecological constraints.This research was funded partially by NSF grant (IBN-0235820) to TWC and Sigma Xi Grants-in-Aid to AGC and by the National Coral Reef Institute through a subaward to PHB and RL Caldwell through the NOAA Coastal Ocean Program under award #NA16OA2413, to Nova Southeastern University

Development of a Fragment-Based in Silico Profiler for Michael Addition Thiol Reactivity

The Adverse Outcome Pathway (AOP) paradigm details the existing knowledge that links the initial interaction between a chemical and a biological system, termed the molecular initiating event (MIE), through a series of intermediate events, to an adverse effect. An important example of a well-defined MIE is the formation of a covalent bond between a biological nucleophile and an electrophilic compound. This particular MIE has been associated with various toxicological end points such as acute aquatic toxicity, skin sensitization, and respiratory sensitization. This study has investigated the calculated parameters that are required to predict the rate of chemical bond formation (reactivity) of a dataset of Michael acceptors. Reactivity of these compounds toward glutathione was predicted using a combination of a calculated activation energy value (Eact, calculated using density functional theory (DFT) calculation at the B3YLP/6-31G+(d) level of theory, and solvent-accessible surface area values (SAS) at the α carbon. To further develop the method, a fragment-based algorithm was developed enabling the reactivity to be predicted for Michael acceptors without the need to perform the time-consuming DFT calculations. Results showed the developed fragment method was successful in predicting the reactivity of the Michael acceptors excluding two sets of chemicals: volatile esters with an extended substituent at the β-carbon and chemicals containing a conjugated benzene ring as part of the polarizing group. Additionally the study also demonstrated the ease with which the approach can be extended to other chemical classes by the calculation of additional fragments and their associated Eact and SAS values. The resulting method is likely to be of use in regulatory toxicology tools where an understanding of covalent bond formation as a potential MIE is important within the AOP paradigm

Read-Across for Rat Oral Gavage Repeated-Dose Toxicity for Short-Chain Mono-Alkylphenols: A Case Study

Short-chain mono-alkylphenols provide an example of where a category-approach to read-across may be used to estimate the repeated-dose endpoint for a number of derivatives. Specifically, the NOAELs of 50 mg/kg bw/d for mono-methylphenols based on a LOAEL of very low systemic toxicity can be read across with confidence to untested mono-alkylphenols in the category. These simple alkylphenols are non-reactive and exhibit an unspecific, reversible polar narcosis mode of toxic action. Briefly, polar narcotics act via unspecific, reversible interactions with biological membranes in a manner similar to cataleptic anaesthetics. The read-across premise includes rapid and complete absorption via the gastrointestinal tract, distribution in the circulatory system, first-pass Phase 2 metabolism in the liver, and elimination of sulphates and glucuronides in the urine. Thus, toxicokinetic parameters are considered to be similar and have the same toxicological significance. Five analogues have high quality experimental oral repeated-dose toxicity data (i.e., OECD TG 408 or OECD TG 422). These repeated-dose toxicity test results exhibit qualitative consistency in symptoms. Typical findings include decreased body weight and slightly increased liver and kidney weights which are generally without concurrent histopathological effects. The sub-chronic findings are quantitatively consistent with the No Observed Adverse Effect Level (NOAEL) of ≥ 50 mg/kg bw/d. Chemical similarity between the analogues is readily defined, and data uncertainty associated with the similarities in toxicokinetic properties, as well as toxicodynamic properties, are low. Uncertainty associated with mechanistic relevance and completeness of the read-across is low-to-moderate, largely because there is no adverse outcome pathway or intermediate event data. Uncertainty associated with mechanistic relevance and completeness of the read-across is reduced by the concordance of in vivo, in vitro, USEPA toxicity forecaster (ToxCast) results, as well as the in silico data. The rat oral repeated-dose NOAEL values for the source substances can be read across to fill the data gaps of the untested analogues in this category with uncertainty deemed equivalent to results from a TG 408 assessment

Validation of a fragment-based profiler for thiol reactivity for the prediction of toxicity: skin sensitisation and tetrahymena pyriformis

This study outlines the use of a recently developed fragment-based thiol reactivity profiler for Michael acceptors to predict toxicity towards Tetrahymena pyriformis and skin sensitisation potency as determined in the Local Lymph Node Assay (LLNA). The results showed that the calculated reactivity parameter from the profiler, -log RC50(calc), was capable of predicting toxicity for both endpoints with excellent statistics. However, the study highlighted the importance of a well-defined applicability domain for each endpoint. In terms of Tetrahymena pyriformis this domain was defined in terms of how fast or slowly a given Michael acceptor reacts with thiol leading to two separate quantitative structure-activity models. The first, for fast reacting chemicals required only –Log RC50(calc) as a descriptor, whilst the second required the addition of a descriptor for hydrophobicity. Modelling of the LLNA required only a single descriptor, -log RC50(calc), enabling potency to be predicted. The applicability domain excluded chemicals capable of undergoing polymerisation and those that were predicted to be volatile. The modelling results for both endpoints, using the –log RC50(calc) value from the profiler, were in keeping with previously published studies that have utilised experimentally determined measurements of reactivity. This results demonstrate the output from the fragment-based thiol reactivity profiler can be used to develop quantitative structure-activity relationship models where reactivity towards thiol is a driver of toxicity

Assessing Uncertainty in Read-Across: Questions to Evaluate Toxicity Predictions Based on Knowledge Gained from Case Studies

Read-across as an alternative assessment method for chemical toxicity has growing interest in both the regulatory and industrial communities. The pivotal means of acquiring acceptance of a read-across prediction is identifying and assessing uncertainties associated with it. This study has identified and summarised in a structured way the variety of uncertainties that potentially impact acceptance of a readacross argument. The main sources of uncertainty were established and divided into four main categories: i) the regulatory use of the prediction, ii) the data for the apical endpoint being assessed, iii) the readacross argumentation, and iv) the similarity justification. Specifically, the context of, and relevance to, the regulatory use of a read-across will dictate the acceptable level of uncertainties. The apical endpoint (or other) data must be of sufficient quality and relevance for data gap filling. Read-Across argumentation uncertainties include: 1) mechanistic plausibility (i.e., the knowledge of the chemical and biological mechanisms leading to toxicity), 2) completeness of the supporting evidence, 3) robustness of the supporting data, and 4) Weight-of-Evidence. In addition, similarity arguments for chemistry, physicochemical properties, toxicokinetics and toxicodynamics are linked to these read-across argumentation issues. To further progress in this area, a series of questions are proposed with the goal of addressing each type of uncertainty

The secret world of shrimps: polarisation vision at its best

Animal vision spans a great range of complexity, with systems evolving to detect variations in optical intensity, distribution, colour, and polarisation. Polarisation vision systems studied to date detect one to four channels of linear polarisation, combining them in opponent pairs to provide intensity-independent operation. Circular polarisation vision has never been seen, and is widely believed to play no part in animal vision. Polarisation is fully measured via Stokes' parameters--obtained by combined linear and circular polarisation measurements. Optimal polarisation vision is the ability to see Stokes' parameters: here we show that the crustacean \emph{Gonodactylus smithii} measures the exact components required. This vision provides optimal contrast-enhancement, and precise determination of polarisation with no confusion-states or neutral-points--significant advantages. We emphasise that linear and circular polarisation vision are not different modalities--both are necessary for optimal polarisation vision, regardless of the presence of strongly linear or circularly polarised features in the animal's environment.Comment: 10 pages, 6 figures, 2 table

On visual pigment templates and the spectral shape of invertebrate rhodopsins and metarhodopsins

The absorbance spectra of visual pigments can be approximated with mathematical expressions using as single parameter the absorbance peak wavelength. A comparison of the formulae of Stavenga et al. in Vision Res 33:1011–1017 (1993) and Govardovskii et al. in Vis Neurosci 17:509–528 (2000) applied to a number of invertebrate rhodopsins reveals that both templates well describe the normalized α-band of rhodopsins with peak wavelength > 400 nm; the template spectra are virtually indistinguishable in an absorbance range of about three log units. The template formulae of Govardovskii et al. in Vis Neurosci 17:509–528 (2000) describe the rhodopsin spectra better for absorbances below 10−3. The template predicted spectra deviate in the ultraviolet wavelength range from each other as well as from measured spectra, preventing a definite conclusion about the spectral shape in the wavelength range <400 nm. The metarhodopsin spectra of blowfly and fruitfly R1-6 photoreceptors derived from measured data appear to be virtually identical. The established templates describe the spectral shape of fly metarhodopsin reasonably well. However, the best fitting template spectrum slightly deviates from the experimental spectra near the peak and in the long-wavelength tail. Improved formulae for fitting the fly metarhodopsin spectra are proposed