Integrating cell screening and mechanism of action data for organometallic anticancer agents

Both acquired and intrinsic drug resistance are established clinical problems in many areas of medicine. This is particularly evident with the growing resistance to platinum chemotherapy agents in cancer treatment programmes. New, alternative treatments for platinum-resistance patients are needed, with comparable potency, no platinum cross- resistance, better safety profiles and which target non-repairable areas of the cell, reducing acquired resistance. This thesis focuses on osmium- and iridium-based organometallic anticancer agents to fill this clinical need. Previous work has validated their potency, safety and activity in platinum-resistant cancers, however, their mechanism of action (MOA) was yet to be identified. Knowing the MOA of new compounds is essential for personalising and stratifying cancer treatment, allowing for better patient selection and prediction of treatment outcomes. Often identifying the biological target of a new therapeutic is not essential. Instead, quantifying the cellular response to that treatment, and identifying cell types which hold beneficial biological properties to optimise compound effects, is more effective. This thesis has applied the principles of systems biology to study the whole cell effect of osmium and iridium compounds in epithelial ovarian cancer. Cells were studied at the transcriptional, translational and structural level to investigate compound response, integrating a selection of these findings using novel statistical modelling. Results propose that these compounds induce oxidative stress in cancer cells, and subsequently damage DNA to exert antiproliferative effects at submicromolar concentrations. This is the first example of studying organometallic compounds using this combination of techniques, and is a promising work flow for future efforts in this area

Water Ice and Dust in the Innermost Coma of Comet 103P/Hartley 2

On November 4th, 2010, the Deep Impact eXtended Investigation (DIXI) successfully encountered comet 103P/Hartley 2, when it was at a heliocentric distance of 1.06 AU. Spatially resolved near-IR spectra of comet Hartley 2 were acquired in the 1.05-4.83 micron wavelength range using the HRI-IR spectrometer. We present spectral maps of the inner ~10 kilometers of the coma collected 7 minutes and 23 minutes after closest approach. The extracted reflectance spectra include well-defined absorption bands near 1.5, 2.0, and 3.0 micron consistent in position, bandwidth, and shape with the presence of water ice grains. Using Hapke's radiative transfer model, we characterize the type of mixing (areal vs. intimate), relative abundance, grain size, and spatial distribution of water ice and refractories. Our modeling suggests that the dust, which dominates the innermost coma of Hartley 2 and is at a temperature of 300K, is thermally and physically decoupled from the fine-grained water ice particles, which are on the order of 1 micron in size. The strong correlation between the water ice, dust, and CO2 spatial distribution supports the concept that CO2 gas drags the water ice and dust grains from the nucleus. Once in the coma, the water ice begins subliming while the dust is in a constant outflow. The derived water ice scale-length is compatible with the lifetimes expected for 1-micron pure water ice grains at 1 AU, if velocities are near 0.5 m/s. Such velocities, about three order of magnitudes lower than the expansion velocities expected for isolated 1-micron water ice particles [Hanner, 1981; Whipple, 1951], suggest that the observed water ice grains are likely aggregates.Comment: 51 pages, 12 figures, accepted for publication in Icaru

The Relationship between Classroom Assessment Practices and Student Motivation and Engagement

Assessment is becoming increasingly important in classrooms as school systems respond to federal and state testing mandates. The increasing emphasis on assessment surely impacts student outcomes such as engagement and motivation, and ultimately achievement. While it is clear that classroom assessment is receiving more attention as a critical component of teaching that directly affects student learning, there is still a lack of systematic research that addresses many classroom assessment issues. There is some evidence that effective formative assessment enhances achievement (Black & Williams 1998), and that certain grading practices result in greater student motivation and achievement (Brookhart, 2004b). However, researchers have not sufficiently investigated how classroom assessment and grading practices should be categorized, and have not demonstrated strong relationships between these practices and student self-efficacy and motivation at different grade levels

Pharmaco-genomic investigations of organo-iridium anticancer complexes reveal novel mechanism of action

Resistance to platinum drugs (used in >50% of cancer chemotherapies) is a clinical problem. Other precious metal complexes with distinct mechanisms of action might overcome this. Half-sandwich organometallic complexes containing arene or cyclopentadienyl (Cp) ligands show promise. We screened two iridium(III) complexes [Ir(CpXbiph)(ppy)Cl] (ZL49, 1, ppy = phenylpyridine) and [Ir(CpXph)(azpyNMe2)Cl]PF6 (ZL109, 2, azpyNMe2 = N,N-dimethylphenylazopyridine) in 916 cancer cell lines from 28 tissue types. On average, complex 2 was 78× more potent than 1, 36× more active than cisplatin (CDDP), and strongly active (nanomolar) in patient-derived ovarian cancer cell lines. RNA sequencing of A2780 ovarian cells revealed upregulation of antioxidant responses (NRF2, AP-1) consistent with observed induction of reactive oxygen species (ROS). Protein microarrays, high content imaging and cell cycle analysis showed S/G2 arrest, and late-stage DNA damage response without p53 requirement. The triple-negative breast cancer cell line OCUB-M was highly sensitive to 2 as were cell lines with KIT mutations. Complex 2 exhibits a markedly different pattern of antiproliferative activity compared to the 253 drugs in the Sanger Cancer Genome database, but is most similar to osmium(II) arene complexes which share the same azopyridine ligand. Redox modulation and DNA damage can provide a multi-targeting strategy, allowing compounds such as 2 to overcome cellular resistance to platinum anticancer drugs

Organometallic iridium(III) anticancer complexes with new mechanisms of action: NCI-60 screening, mitochondrial targeting, and apoptosis

Platinum complexes related to cisplatin, cis-[PtCl2(NH3)2], are successful anticancer drugs; however, other transition metal complexes offer potential for combating cisplatin resistance, decreasing side effects, and widening the spectrum of activity. Organometallic half-sandwich iridium (IrIII) complexes [Ir(Cpx)(XY)Cl]+/0 (Cpx = biphenyltetramethylcyclopentadienyl and XY = phenanthroline (1), bipyridine (2), or phenylpyridine (3)) all hydrolyze rapidly, forming monofunctional G adducts on DNA with additional intercalation of the phenyl substituents on the Cpx ring. In comparison, highly potent complex 4 (Cpx = phenyltetramethylcyclopentadienyl and XY = N,N-dimethylphenylazopyridine) does not hydrolyze. All show higher potency toward A2780 human ovarian cancer cells compared to cisplatin, with 1, 3, and 4 also demonstrating higher potency in the National Cancer Institute (NCI) NCI-60 cell-line screen. Use of the NCI COMPARE algorithm (which predicts mechanisms of action (MoAs) for emerging anticancer compounds by correlating NCI-60 patterns of sensitivity) shows that the MoA of these IrIII complexes has no correlation to cisplatin (or oxaliplatin), with 3 and 4 emerging as particularly novel compounds. Those findings by COMPARE were experimentally probed by transmission electron microscopy (TEM) of A2780 cells exposed to 1, showing mitochondrial swelling and activation of apoptosis after 24 h. Significant changes in mitochondrial membrane polarization were detected by flow cytometry, and the potency of the complexes was enhanced ca. 5× by co-administration with a low concentration (5 μM) of the γ-glutamyl cysteine synthetase inhibitor L-buthionine sulfoximine (L-BSO). These studies reveal potential polypharmacology of organometallic IrIII complexes, with MoA and cell selectivity governed by structural changes in the chelating ligands

The benefits and challenges of using crowdfunding to facilitate community-led projects in the context of digital civics

Digital technology is increasingly being used to bring citizens and communities together to address local concerns. While a variety of approaches have been developed that allow citizens and communities to improve their local communities, these approaches are often financially unsustainable. In this paper, we describe our exploration of crowdfunding as an alternative approach to funding and sustaining community-led projects in the context of digital civics. Through our analysis of four community-led crowdfunding projects, we explore the merits of crowdfunding in this context, demonstrating that it can a) provide an alternative funding mechanism suitable for financing some community-led projects, b) create a sense of empowerment and ownership, and c) increase community awareness. By reflecting on our experiences, we identify four key challenges to utilising crowdfunding to support community-led projects in the context digital civics. We also provide advice specific to crowdfunding in the context of digital civics, before discussing the role of crowdfunding within digital civics. By addressing these challenges, we will be able to better support community groups crowdfund for the public good

Global collision-risk hotspots of marine traffic and the world’s largest fish, the whale shark

© The Author(s), 2022. This article is distributed under the terms of the Creative Commons Attribution License. The definitive version was published in Womersley, F. C., Humphries, N. E., Queiroz, N., Vedor, M., da Costa, I., Furtado, M., Tyminski, J. P., Abrantes, K., Araujo, G., Bach, S. S., Barnett, A., Berumen, M. L., Bessudo Lion, S., Braun, C. D., Clingham, E., Cochran, J. E. M., de la Parra, R., Diamant, S., Dove, A. D. M., Dudgeon, C. L., Erdmann, M. V., Espinoza, E., Fitzpatrick, R., González Cano, J., Green, J. R., Guzman, H. M., Hardenstine, R., Hasan, A., Hazin, F. H. V., Hearn, A. R., Hueter, R. E., Jaidah, M. Y., Labaja, J., Ladinol, F., Macena, B. C. L., Morris Jr., J. J., Norman, B. M., Peñaherrera-Palmav, C., Pierce, S. J., Quintero, L. M., Ramırez-Macías, D., Reynolds, S. D., Richardson, A. J., Robinson, D. P., Rohner, C. A., Rowat, D. R. L., Sheaves, M., Shivji, M. S., Sianipar, A. B., Skomal, G. B., Soler, G., Syakurachman, I., Thorrold, S. R., Webb, D. H., Wetherbee, B. M., White, T. D., Clavelle, T., Kroodsma, D. A., Thums, M., Ferreira, L. C., Meekan, M. G., Arrowsmith, L. M., Lester, E. K., Meyers, M. M., Peel, L. R., Sequeira, A. M. M., Eguıluz, V. M., Duarte, C. M., & Sims, D. W. Global collision-risk hotspots of marine traffic and the world’s largest fish, the whale shark. Proceedings of the National Academy of Sciences of the United States of America, 119(20), (2022): e2117440119, https://doi.org/10.1073/pnas.2117440119.Marine traffic is increasing globally yet collisions with endangered megafauna such as whales, sea turtles, and planktivorous sharks go largely undetected or unreported. Collisions leading to mortality can have population-level consequences for endangered species. Hence, identifying simultaneous space use of megafauna and shipping throughout ranges may reveal as-yet-unknown spatial targets requiring conservation. However, global studies tracking megafauna and shipping occurrences are lacking. Here we combine satellite-tracked movements of the whale shark, Rhincodon typus, and vessel activity to show that 92% of sharks’ horizontal space use and nearly 50% of vertical space use overlap with persistent large vessel (>300 gross tons) traffic. Collision-risk estimates correlated with reported whale shark mortality from ship strikes, indicating higher mortality in areas with greatest overlap. Hotspots of potential collision risk were evident in all major oceans, predominantly from overlap with cargo and tanker vessels, and were concentrated in gulf regions, where dense traffic co-occurred with seasonal shark movements. Nearly a third of whale shark hotspots overlapped with the highest collision-risk areas, with the last known locations of tracked sharks coinciding with busier shipping routes more often than expected. Depth-recording tags provided evidence for sinking, likely dead, whale sharks, suggesting substantial “cryptic” lethal ship strikes are possible, which could explain why whale shark population declines continue despite international protection and low fishing-induced mortality. Mitigation measures to reduce ship-strike risk should be considered to conserve this species and other ocean giants that are likely experiencing similar impacts from growing global vessel traffic.Funding for data analysis was provided by the UK Natural Environment Research Council (NERC) through a University of Southampton INSPIRE DTP PhD Studentship to F.C.W. Additional funding for data analysis was provided by NERC Discovery Science (NE/R00997/X/1) and the European Research Council (ERC-AdG-2019 883583 OCEAN DEOXYFISH) to D.W.S., Fundação para a Ciência e a Tecnologia (FCT) under PTDC/BIA/28855/2017 and COMPETE POCI-01–0145-FEDER-028855, and MARINFO–NORTE-01–0145-FEDER-000031 (funded by Norte Portugal Regional Operational Program [NORTE2020] under the PORTUGAL 2020 Partnership Agreement, through the European Regional Development Fund–ERDF) to N.Q. FCT also supported N.Q. (CEECIND/02857/2018) and M.V. (PTDC/BIA-COM/28855/2017). D.W.S. was supported by a Marine Biological Association Senior Research Fellowship. All tagging procedures were approved by institutional ethical review bodies and complied with all relevant ethical regulations in the jurisdictions in which they were performed. Details for individual research teams are given in SI Appendix, section 8. Full acknowledgments for tagging and field research are given in SI Appendix, section 7. This research is part of the Global Shark Movement Project (https://www.globalsharkmovement.org)

Diving into the vertical dimension of elasmobranch movement ecology

Knowledge of the three-dimensional movement patterns of elasmobranchs is vital to understand their ecological roles and exposure to anthropogenic pressures. To date, comparative studies among species at global scales have mostly focused on horizontal movements. Our study addresses the knowledge gap of vertical movements by compiling the first global synthesis of vertical habitat use by elasmobranchs from data obtained by deployment of 989 biotelemetry tags on 38 elasmobranch species. Elasmobranchs displayed high intra- and interspecific variability in vertical movement patterns. Substantial vertical overlap was observed for many epipelagic elasmobranchs, indicating an increased likelihood to display spatial overlap, biologically interact, and share similar risk to anthropogenic threats that vary on a vertical gradient. We highlight the critical next steps toward incorporating vertical movement into global management and monitoring strategies for elasmobranchs, emphasizing the need to address geographic and taxonomic biases in deployments and to concurrently consider both horizontal and vertical movements

Diving into the vertical dimension of elasmobranch movement ecology

Knowledge of the three-dimensional movement patterns of elasmobranchs is vital to understand their ecological roles and exposure to anthropogenic pressures. To date, comparative studies among species at global scales have mostly focused on horizontal movements. Our study addresses the knowledge gap of vertical movements by compiling the first global synthesis of vertical habitat use by elasmobranchs from data obtained by deployment of 989 biotelemetry tags on 38 elasmobranch species. Elasmobranchs displayed high intra- and interspecific variability in vertical movement patterns. Substantial vertical overlap was observed for many epipelagic elasmobranchs, indicating an increased likelihood to display spatial overlap, biologically interact, and share similar risk to anthropogenic threats that vary on a vertical gradient. We highlight the critical next steps toward incorporating vertical movement into global management and monitoring strategies for elasmobranchs, emphasizing the need to address geographic and taxonomic biases in deployments and to concurrently consider both horizontal and vertical movements