Background: Midostaurin is approved for FLT3 mutant-positive (FLT3+) acute myeloid leukemia (AML), however efficacy has also been observed in a subpopulation of FLT3 mutant-negative AML, suggesting that FLT3 mutation is not the only determinant conferring midostaurin sensitivity.Casado et al previously described phosphoprotein signatures significantly associated with ex vivo responses to midostaurin in primary AML blasts (Casado et al Leukaemia 2018). In the current study, we tested whether our signatures could group FLT3+ patients based on clinical responses to midostaurin plus chemotherapy.Methods: FLT3+ bone marrow (BM) and peripheral blood (PB) specimens were collected at diagnosis, post-treatment and relapse (n=54 cases) from the Leukemia Tissue Bank at Princess Margaret Cancer Centre. All patients in this study were treated with standard chemotherapy plus midostaurin. Protein/phosphoprotein-signatures for BM and PB samples were analysed independently. Case-studies with multiple post-treatment time-points or relapse events following second line treatments were also analysed.Peptides (proteomics) and enriched phosphopeptides (phosphoproteomics) were quantified using liquid chromatography - tandem mass spectrometry. A classification machine learning (ML) algorithm was trained to group patients based on response to treatment as a function of protein/phosphoprotein-signature status. Other features (e.g. genetic mutations, HSC-transplant) were also analysed. Differential survival analysis between patient groups was carried out with Kaplan-Meier and Log Rank test methods.Pathways upregulated in post-treatment or relapse specimens, particularly from those cases that responded poorly to chemo + midostaurin (i.e. early relapse / refractory disease) were investigated using enrichment statistical methods including kinase-substrate enrichment analysis (KSEA) and gene ontology analysis and identified as potential mechanisms of resistance. Statistical significance of enrichment was determined using parametric methods and p-values adjusted for multiple testing using the Benjamini-Hochberg method.Results: ML models were developed based on the ex-vivo phosphoproteomics signatures described in the Casado et al study, from which we trained a predictive model (model 1). Patients positive for model 1 exhibited a survival probability of 243 weeks, compared to 126 weeks in signature negative patients (averages by geometric mean, Log Rank p = 9.88e-05).As the patients in the current study received chemotherapy, in addition to midostaurin, we identified a new phosphoproteomic signature consisting of 26 phospho-sites which partially overlapped with the ex-vivo signature. Patients positive for this new phosphoproteomic signature showed a markedly longer survival time than negative patients (269 vs 76 weeks, Log Rank p = 1.30e-05 for PB and 241 vs 56, Log Rank p = 2.13e-09 for BM specimens, Table).A proteomic signature was also identified in the current study. Positive patients showed a longer survival time than negative patients (330 vs 173 weeks, Log Rank p = 5.0e-04 for PB and 460 vs 156, Log Rank p = 5.2e-06 for BM specimens, Table), however this was less differentiating than the phosphoproteomic signature.Pathways upregulated in post-treatment or relapse specimens from early relapse or refractory cases were associated with molecular functions such as cell proliferation, anti-apoptosis, non-homologous end-joining, transcriptional regulation, spliceosome and cytoskeleton remodelling.Conclusions: We have identified protein and phosphoprotein signatures with the potential to further stratify AML for midostaurin treatment. Phosphoproteomic signatures differentiated according to response better than the proteomic signatures. Pathways upregulated in relapse/refractory cases may have a role in resistance and this will be determined in follow up studies. Analysis will also be performed on FLT3 mutant-negative cases to validate the signatures and elucidate mechanisms of resistance in this group. DisclosuresVeiga Nobre: Kinomica Ltd.: Current Employment. Minden: Astellas: Consultancy. Gribben: Janssen: Honoraria, Research Funding; AZ: Honoraria, Research Funding; Abbvie: Honoraria; BMS: Honoraria; Gilead/Kite: Honoraria; Morphosys: Honoraria; Novartis: Honoraria; Takeda: Honoraria; TG Therapeutics: Honoraria. Britton: Kinomica Ltd.: Current Employment, Current equity holder in publicly-traded company.

<h3>Introduction</h3> Corporate governance, through the involvement of employees and the dynamics that it can produce, is an essential issue for efficiency and for safeguarding workers’ health. Many studies have shown that individual autonomy is a determining factor for occupational health. Employees must have individual and collective leeway in organising their work, not only applying procedures that have been defined by others but being able to show their creativity and influence their work environment. In this aim, organisational design should provide the opportunity to influence management modes, distribution of power, individual participation in the design processes. The concept of subsidiarity can help to achieve these goals. <h3>Methods</h3> From an ergonomic intervention in an insurance company about the organisation of a call centre, we can see what are the conditions to build an organisation aiming at subsidiarity, both as a target and as a process for the management of change. <h3>Results</h3> Contrary to the idea of a predetermined definition of all the decisions that can be taken at every level of the hierarchy, efficiency consists in constantly adapting the level of decision-making to the problem being dealt with. This requires constructing an organisation which is sensitive to details of events, which can be always adjusting the levels at which matters are dealt with. However, this swing from one decision level to another is only possible if the organisation and the people within it are well prepared beforehand. <h3>Conclusion</h3> The concept of subsidiarity constitutes an ethical point of reference for dealing with questions of organisational design and corporate governance. It favours decision-making at the lowest appropriate level by following three organising principles: the competency principle, the assistance principle, and the substitution principle. The way we build the interventions must be in line with this vision. It is both a matter of efficiency and a dynamic for healthy organisations.

The AR-CALUX® in vitro method is a reporter gene-based transactivation method where endocrine active chemicals with androgenic or anti-androgenic potential can be detected. Its primary purpose is for screening chemicals for further prioritization and providing mechanistic (endocrine mode of action) information, as defined by the Organisation of Economic Cooperation and Development (OECD) conceptual framework for the testing and assessment of endocrine-disrupting chemicals. This article describes the conduct and results of an international ring trial with 3 EU-NETVAL laboratories and the test method developer. It was organized by EURL ECVAM to validate the method by testing 46 chemicals. A very good reproducibility within and between laboratories was concluded (94.7–100% and 100% concordance of classification) with low within and between laboratory variability (less than 2.5% CV on EC50 values). Moreover, the variability is within the range of other validated, mechanistically similar methods. In comparison to the AR-reference list compiled by ICCVAM, an almost 100% concordance of classifications was obtained. This method allows the detection of the agonist and antagonist properties of a chemical. A specificity control test was developed during the validation study and added to the antagonist assay rendering the assay more specific. A comparison is made with the mechanistically similar methods AR-EcoScreen™ and 22Rv1/MMTV GR-KO TA. The AR-CALUX® method was approved for inclusion in the recently updated OECD test guideline TG458 which incorporates all 3 methods.

In response to the careers article “On the road less travelled” (December 2020 pp45–46), in which Tamia Williams from Pace University in New York discusses her experiences as a Black physicist.

Methods for residue-selective and stable modification of canonical amino acids enable the installation of distinct functionality which can aid in the interrogation of biological processes or the generation of new therapeutic modalities. Herein, we report an extensive investigation of reactivity and stability profiles for a series of vinylheteroarene motifs. Studies on small molecule and protein substrates identified an optimum vinylheteroarene scaffold for selective cysteine modification. Utilisation of this lead linker to modify a number of protein substrates with various functionalities, including the synthesis of a homogeneous, stable and biologically active antibody–drug conjugate (ADC) was then achieved. The reagent was also efficient in labelling proteome-wide cysteines in cell lysates. The efficiency and selectivity of these reagents as well as the stability of the products makes them suitable for the generation of biotherapeutics or studies in chemical biology.

In this perspective we deal with questions pertaining to the development of synthesis planning technologies over the course of recent years.

In response to Stephen Ornes’ feature “How to hack a self-driving car” (August 2020) in which he looks into the hardware and software behind cars that drive themselves, and the potential opportunities they create for hackers.

The urban population will rise to 6.7 billion by 2050. The United Nations has committed to provide everyone with safely managed sanitation, but there is limited understanding of the scale of the challenge. This paper describes a methodology for rapid assessment of sanitation in cities including a graphical representation (a shit-flow diagram or SFD) and reports on findings from implementation in 39 cities. The SFD provides high level information for planning purposes covering the entire sanitation system in a city. More than half of the human excreta produced in these cities is not safely managed. The most significant portions of the unsafely managed excreta are: (i) contents of pits and tanks which are not emptied and are overflowing, leaking, or discharging to the surrounding environment (14%); (ii) contents of pits and tanks which are emptied but not delivered to treatment (18%); (iii) fecal sludge and supernatant delivered to treatment but not treated (3%); (iv) wastewater in sewers not delivered to treatment (14%); and (v) wastewater delivered to treatment but not treated (6%). Many cities currently relying on onsite sanitation for safe storage, particularly in Africa, will need new strategies as populations grow. Containment systems that discharge to open drains are common in some Asian cities; these pose a public health risk. Dumping of excreta is widespread and there is a lack of realistic performance data on which estimates of the extent and effectiveness of treatment can be made. The SFD production process can be challenging due to a lack of data and low technical capacity in cities. There is often uncertainty over terminology and over the status of infrastructure. Formalizing definitions for the SFD preparation process was found to be useful in overcoming capacity constraints in cities. The SFD produces a credible snapshot of the sanitation situation in a city. The paper provides evidence of the urgent need for improved management and monitoring of urban sanitation in cities around the world and highlights the role of the SFD as a planning tool.

The development of a continuous flow ruthenium-catalyzed ester reduction using hydrogen gas is described. The assessment of green metrics show the protocol is substantially more sustainable than commonly employed metal hydride reductions.

Building on our previously-reported novel tricyclic topoisomerase inhibitors (NTTIs), we disclose the discovery of REDX07965, which has an MIC90 of 0.5 μg mL-1 against Staphylococcus aureus, favourable in vitro pharmacokinetic properties, selectivity versus human topoisomerase II and an acceptable toxicity profile. The results herein validate a rational design approach to address the urgent unmet medical need for novel antibiotics.