Development of in silico models to support repeat dose safety assessment of cosmetic ingredients to humans

Cosmetic products are used daily on a global scale. Therefore, it is necessary to ensure that these products, and their ingredients, do not cause any adverse human health effects under normal usage; to ensure this, risk assessment must be performed. Traditionally, risk assessments are performed in vivo, i.e. conducting tests on animals using the chemical(s) of interest. However, over the past decade there has been an increase in research into the use of alternative toxicity testing methods, such as in vitro, in chemico and in silico. Whilst there are a number of alternative techniques that may be employed, no one method can be used in isolation as a full replacement for an in vivo test. Therefore, the Adverse Outcome Pathway (AOP) concept is an emerging method by which information provided by the in vitro, in chemico, and in silico approaches can be utilised in an integrated testing strategy. The AOP concept links an upstream molecular initiating event to a downstream adverse outcome, via a number of testable key events.In silico approaches utilise computers in order to develop predictive models. Within the AOP paradigm in silico method work to identify the key features of a chemical (structural alerts) that induce a molecular initiating event (MIE). A collection of structural alerts that induce the same MIE are considered to be an in silico profiler. Typically, these in silico profilers are supported by associated toxicity, or mechanistic, information pertaining to the ability to induce a specific MIE. The overall aim of the work presented in this thesis was the development of an in silico profiler, based upon the hypothesis that the induction of mitochondrial toxicity is a key driver of organ-level toxicity. The research presented herein demonstrates the ability to identify, and develop, two types of structural alert; mechanism- and chemistry-based; that pertain to mitochondrial toxicity. Due to the differences inherent in these two types of alert they should be utilised for different purposes. As such, the main usage of the mechanism-based alerts should be in the formation of chemical categories and subsequent data gap filling via read-across. In comparison, the chemistry-based alerts should be utilised for the purposes of prioritising chemicals, within an inventory, that should undergo additional testing in in vitro and/or in chemico assays. It is envisaged that these two types of structural alerts could be used to profile chemical inventories as part of a tiered testing strategy.Therefore, the future work discussed in detail the need to expand the chemical space covered by the alerts. Additional future work involves utilising experimental information from in vitro/in chemico assays to verify the mechanism-based alerts and to refine the chemistry-based alerts by discerning mechanistic information associated with them. Furthermore, it is envisaged that these alerts could be incorporated into predictive tools, such as the OECD QSAR Toolbox, to enable their use for screening and prioritisation purposes

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

IL-4 receptor-alpha-dependent control of Cryptococcus neoformans in the early phase of pulmonary infection

Cryptococcus neoformans is an opportunistic fungal pathogen that causes lung inflammation and meningoencephalitis in immunocompromised people. Previously we showed that mice succumb to intranasal infection by induction of pulmonary interleukin (IL)-4Rα-dependent type 2 immune responses, whereas IL-12-dependent type 1 responses confer resistance. In the experiments presented here, IL-4Rα −/− mice unexpectedly show decreased fungal control early upon infection with C. neoformans , whereas wild-type mice are able to control fungal growth accompanied by enhanced macrophage and dendritic cell recruitment to the site of infection. Lower pulmonary recruitment of macrophages and dendritic cells in IL-4Rα −/− mice is associated with reduced pulmonary expression of CCL2 and CCL20 chemokines. Moreover, IFN-γ and nitric oxide production are diminished in IL-4Rα −/− mice compared to wild-type mice. To directly study the potential mechanism(s) responsible for reduced production of IFN-γ, conventional dendritic cells were stimulated with C. neoformans in the presence of IL-4 which results in increased IL-12 production and reduced IL-10 production. Together, a beneficial role of early IL-4Rα signaling is demonstrated in pulmonary cryptococcosis, which contrasts with the well-known IL-4Rα-mediated detrimental effects in the late phase

Physical therapists collect different outcome measures after total joint arthroplasty as compared to most orthopaedic surgeons: a New England study

Background: Following total knee and hip arthroplasty, patient progress can be assessed with patient-reported outcome measures (PROMs) and performance-based outcome measures (PBOMs). The American Joint Replacement Registry 2016 guide recommends collecting several measures, including Patient Reported Outcome Measure Information System Global, Knee Injury and Osteoarthritis Outcome Score Jr, and Hip Injury and Osteoarthritis Outcome Score Jr. This study aimed to assess the current and anticipated use of PROMs and PBOMs by New England physical therapists. Methods: An online survey was conducted in July and August of 2015 asking physical therapists in New England to rate their current and anticipated future use of PROMs and PBOMs in terms of clinical decision making associated with the treatment and care of patients after total hip and knee replacement. Results: There were 122 responses. The most often used and recommended PROMS were the Numeric Pain Rating Scale (99.2% and 97.5%, respectively) and Lower Extremity Function Scale (76.2% and 77.0%). There was significant variability in the use of different PBOMs, but the most often used and recommended were the Timed Up and Go (93.4% and 85.2%) and the Single Leg Balance Test (90.2% and 87.7%). Conclusions: This study suggests that orthopaedic surgeons and physical therapists use different PROMs and PBOMs for postoperative assessment of total joint patients and highlights the need for more collaboration and consistency between these disciplines. Keywords: Outcome measures, Knee arthroplasty, Hip arthroplasty, Patient-reported outcome measures, Rehabilitatio

IL-4 determines eicosanoid formation in dendritic cells by down-regulation of 5-lipoxygenase and up-regulation of 15-lipoxygenase 1 expression

Dendritic cell (DC) differentiation from human CD34(+) hematopoietic progenitor cells (HPCs) can be triggered in vitro by a combination of cytokines consisting of stem cell factor, granulocyte-macrophage colony-stimulating factor, and tumor necrosis factor α. The immune response regulatory cytokines, IL-4 and IL-13, promote DC maturation from HPCs, induce monocyte-DC transdifferentiation, and selectively up-regulate 15-lipoxygenase 1 (15-LO-1) in blood monocytes. To gain more insight into cytokine-regulated eicosanoid production in DCs we studied the effects of IL-4/IL-13 on LO expression during DC differentiation. In the absence of IL-4, DCs that had been generated from CD34(+) HPCs in response to stem cell factor/granulocyte-macrophage colonystimulating factor/tumor necrosis factor α expressed high levels of 5-LO and 5-LO activating protein. However, a small subpopulation of eosinophil peroxidase(+) (EOS-PX) cells significantly expressed 15-LO-1. Addition of IL-4 to differentiating DCs led to a marked and selective down-regulation of 5-LO but not of 5-LO activating protein in DCs and in EOS-PX(+) cells and, when added at the onset of DC differentiation, also prevented 5-LO up-regulation. Similar effects were observed during IL-4- or IL-13-dependent monocyte-DC transdifferentiation. Down-regulation of 5-LO was accompanied by up-regulation of 15-LO-1, yielding 15-LO-1(+) 5-LO-deficient DCs. However, transforming growth factor β1 counteracted the IL-4-dependent inhibition of 5-LO but only minimally affected 15-LO-1 up-regulation. Thus, transforming growth factor β1 plus IL-4 yielded large mature DCs that coexpress both LOs. Localization of 5-LO in the nucleus and of 15-LO-1 in the cytosol was maintained at all cytokine combinations in all DC phenotypes and in EOS-PX(+) cells. In the absence of IL-4, major eicosanoids of CD34(+)-derived DCs were 5S-hydroxyeicosatetraenoic acid (5S-HETE) and leukotriene B(4), whereas the major eicosanoids of IL-4-treated DCs were 15S-HETE and 5S-15S-diHETE. These actions of IL-4/IL-13 reveal a paradigm of eicosanoid formation consisting of the inhibition of one and the stimulation of another LO in a single leukocyte lineage