Application of in silico and in vitro methods in the development of adverse outcome pathway constructs in wildlife

There is a long history of using both in silico and in vitro methods to predict adverse effects in humans and environmental species where toxicity data are lacking. Currently, there is a great deal of interest in applying these methods to the development of so-called ‘adverse outcome pathway’ (AOP) constructs. The AOP approach provides a framework for organizing information at the chemical and biological level, allowing evidence from both in silico and in vitro studies to be rationally combined to fill gaps in knowledge concerning toxicological events. Fundamental to this new paradigm is a greater understanding of the mechanisms of toxicity and, in particular, where these mechanisms may be conserved across taxa, such as between model animals and related wild species. This presents an opportunity to make predictions across diverse species, where empirical data are unlikely to become available as is the case for most species of wildlife

Linking existing in vitro dermal absorption data to physicochemical properties: Contribution to the design of a weight-of-evidence approach for the safety evaluation of cosmetic ingredients with low dermal bioavailability.

To characterize the risk of cosmetic ingredients when threshold toxicity is assumed, often the "margin of safety" (MoS) is calculated. This uncertainty factor is based on the systemic no observable (adverse) effect level (NO(A)EL) which can be derived from in vivo repeated dose toxicity studies. As in vivo studies for the purpose of the cosmetic legislation are no longer allowed in Europe and a validated in vitro alternative is not yet available, it is no longer possible to derive a NO(A)EL value for a new cosmetic ingredient. Alternatively, cosmetic ingredients with a low dermal bioavailability might not need repeated dose data, as internal exposure will be minimal and systemic toxicity might not be an issue. This study shows the possibility of identifying compounds suspected to have a low dermal bioavailability based on their physicochemical properties (molecular weight, melting point, topological polar surface area and log P) and their in vitro dermal absorption data. Although performed on a limited number of compounds, the study suggests a strategic opportunity to support the safety assessor's reasoning to omit a MoS calculation and to focus more on local toxicity and mutagenicity/genotoxicity for ingredients for which limited systemic exposure is to be expected

Non-monotonous crossover between capillary condensation and interface localisation/delocalisation transition in binary polymer blends

Within self-consistent field theory we study the phase behaviour of a symmetric binary AB polymer blend confined into a thin film. The film surfaces interact with the monomers via short range potentials. One surface attracts the A component and the corresponding semi-infinite system exhibits a first order wetting transition. The surface interaction of the opposite surface is varied as to study the crossover from capillary condensation for symmetric surface fields to the interface localisation/delocalisation transition for antisymmetric surface fields. In the former case the phase diagram has a single critical point close to the bulk critical point. In the latter case the phase diagram exhibits two critical points which correspond to the prewetting critical points of the semi-infinite system. The crossover between these qualitatively different limiting behaviours occurs gradually, however, the critical temperature and the critical composition exhibit a non-monotonic dependence on the surface field.Comment: to appear in Europhys.Let

Proposal of an in silico profiler for categorisation of repeat dose toxicity data of hair dyes

This study outlines the analysis of repeat dose toxicity data taken from Scientific Committee on Consumer Safety (SCCS) opinions for commonly used hair dyes in the European Union. Structural similarity was applied to group these chemicals into categories. Subsequent mechanistic analysis suggested that toxicity to mitochondria is potentially a key driver of repeat dose toxicity for chemicals within each of the categories. The mechanistic hypothesis allowed for an in silico profiler consisting of mechanism-based structural alerts to be proposed. This in silico profiler is intended for grouping chemicals into mechanism-based categories within the Adverse Outcome Pathway paradigm

Hypoxia in grape berries: the role of seed respiration and lenticels on the berry pedicel and the possible link to cell death

Advance Access publication 6 March 2018Mesocarp cell death (CD) during ripening is common in berries of seeded Vitis vinifera L wine cultivars. We examined if hypoxia within berries is linked to CD. Internal oxygen concentration ([O₂]) across the mesocarp was measured in berries from Chardonnay and Shiraz, both seeded, and Ruby Seedless, using an oxygen micro-sensor. Steep [O₂] gradients were observed across the skin and [O₂] decreased toward the middle of the mesocarp. As ripening progressed the minimum [O₂] approached zero in the seeded cultivars and correlated to the profile of CD across the mesocarp. Seed respiration declined during ripening, from a large proportion of total berry respiration early to negligible at latter stages. [O₂] increased towards the central axis corresponding to the presence of air spaces visualised using x-ray micro-CT. These air spaces connect to the pedicel where lenticels are located that are critical for berry O₂ uptake as a function of temperature, and when blocked caused hypoxia in Chardonnay berries, ethanol accumulation and CD. The implications of hypoxia in grape berries are discussed in terms of its role in CD, ripening and berry water relations.Zeyu Xiao, Suzy Y. Rogiers, Victor O. Sadras and Stephen D. Tyerma

Effect of Gravity and Confinement on Phase Equilibria: A Density Matrix Renormalization Approach

The phase diagram of the 2D Ising model confined between two infinite walls and subject to opposing surface fields and to a bulk "gravitational" field is calculated by means of density matrix renormalization methods. In absence of gravity two phase coexistence is restricted to temperatures below the wetting temperature. We find that gravity restores the two phase coexistence up to the bulk critical temperature, in agreement with previous mean-field predictions. We calculate the exponents governing the finite size scaling in the temperature and in the gravitational field directions. The former is the exponent which describes the shift of the critical temperature in capillary condensation. The latter agrees, for large surface fields, with a scaling assumption of Van Leeuwen and Sengers. Magnetization profiles in the two phase and in the single phase region are calculated. The profiles in the single phase region, where an interface is present, agree well with magnetization profiles calculated from a simple solid-on-solid interface hamiltonian.Comment: 4 pages, RevTeX and 4 PostScript figures included. Final version as published. To appear in Phys. Rev. Let

Vascular connections into the grape berry: the link of structural investment to seededness

Vascular bundles in the grape pedicel and berry contain the conduits, phloem and xylem, for transport of water, sugar, nutrients and signals into and through the grape berry and play a critical role in berry growth and composition. Here, we assess the vascular anatomy within the proximal region of the berry. Guided using a 3D berry model generated by micro-CT, differential staining of transverse sections of berries and receptacles was followed by fluorescent microscopy. Morphometric and vascular characteristics were analyzed within the central proximal region (brush zone, a fibrous extension from the pedicel vascular system into the berry) of the seeded cultivars Shiraz and Sauvignon Blanc, as well as the stenospermocarpic cultivars Ruby Seedless and Flame Seedless. Observations revealed a change in vascular arrangement from the receptacle into the berry brush zone and differences in xylem element size as well as xylem and phloem area relationships. Xylem anatomical and derived hydraulic parameters, as well as total tissue area of xylem and phloem varied between cultivars and in receptacle and berry components. Variation in vascular growth between grape pedicels and berries was independent of seededness. Differences in receptacle xylem vessel size and distribution could contribute to cultivar-dependent xylem backflow constraint.Zeyu Xiao, Sabrina Chin, Rosemary G. White, Aude M. Gourieroux, Vinay Pagay, Stephen D. Tyerman, Leigh M. Schmidtke and Suzy Y. Rogier

Co-culture of primary rat hepatocytes with rat liver epithelial cells enhances interleukin-6-induced acute-phase protein response

Three different primary rat hepatocyte culture methods were compared for their ability to allow the secretion of fibrinogen and albumin under basal and IL-6-stimulated conditions. These culture methods comprised the co-culture of hepatocytes with rat liver epithelial cells (CC-RLEC), a collagen type I sandwich culture (SW) and a conventional primary hepatocyte monolayer culture (ML). Basal albumin secretion was most stable over time in SW. Fibrinogen secretion was induced by IL-6 in all cell culture models. Compared with ML, CC-RLEC showed an almost three-fold higher fibrinogen secretion under both control and IL-6-stimulated conditions. Induction of fibrinogen release by IL-6 was lowest in SW. Albumin secretion was decreased after IL-6 stimulation in both ML and CC-RLEC. Thus, cells growing under the various primary hepatocyte cell culture techniques react differently to IL-6 stimulation with regard to acute-phase protein secretion. CC-RLEC is the preferred method for studying cytokine-mediated induction of acute-phase proteins, because of the pronounced stimulation of fibrinogen secretion upon IL-6 exposure under these conditions

Histone deacetylase inhibitors valproate and trichostatin A are toxic to neuroblastoma cells and modulate cytochrome P450 1A1, 1B1 and 3A4 expression in these cells

Histone deacetylase inhibitors such as valproic acid (VPA) and trichostatin A (TSA) were shown to exert antitumor activity. Here, the toxicity of both drugs to human neuroblastoma cell lines was investigated using MTT test, and IC50 values for both compounds were determined. Another target of this work was to evaluate the effects of both drugs on expression of cytochrome P450 (CYP) 1A1, 1B1 and 3A4 enzymes, which are known to be expressed in neuroblastoma cells. A malignant subset of neuroblastoma cells, so-called N-type cells (UKF-NB-3 cells) and the more benign S-type neuroblastoma cells (UKF-NB-4 and SK-N-AS cell lines) were studied from both two points of view. VPA and TSA inhibited the growth of neuroblastoma cells in a dose-dependent manner. The IC50 values ranging from 1.0 to 2.8 mM and from 69.8 to 129.4 nM were found for VPA and TSA, respectively. Of the neuroblastoma tested here, the N-type UKF-NB-3 cell line was the most sensitive to both drugs. The different effects of VPA and TSA were found on expression of CYP1A1, 1B1 and 3A4 enzymes in individual neuroblastoma cells tested in the study. Protein expression of all these CYP enzymes in the S-type SK-N-AS cell line was not influenced by either of studied drugs. On the contrary, in another S-type cell line, UKF-NB-4, VPA and TSA induced expression of CYP1A1, depressed levels of CYP1B1 and had no effect on expression levels of CYP3A4 enzyme. In the N-type UKF-NB-3 cell line, the expression of CYP1A1 was strongly induced, while that of CYP1B1 depressed by VPA and TSA. VPA also induced the expression of CYP3A4 in this neuroblastoma cell line

Gradient critical phenomena in the Ising quantum chain: surface behaviour

We consider the influence of a power-law deviation from the critical coupling such that the system is critical at its surface. We develop a scaling theory showing that such a perturbation introduces a new length scale which governs the scaling behaviour of the density profiles as well as the finite-size behaviour of the surface properties. Exact results are obtained for the Ising quantum chain when the perturbation varies linearly whereas the quadratic perturbation is mainly studied numerically. The scaling theory is well confirmed in both cases