The overall cost of transfusion therapy is more influenced by the cost of the product than the cost of providing the transfusion. Depending on the cost adjustment by the supplier for different doses of PLTs, a low-dose transfusion strategy can be less costly.

1Human rhinoviruses express 2 cysteine proteases, 2A and 3C, that are responsible for 2 viral polyprotein processing. Both proteases also suppress host gene expression by 3 inhibiting mRNA transcription, nuclear export and cap-dependent translation. 4However, the relative contribution that each makes in achieving this goal remains 5 Methods 83Cell lines and reagents 84 293T cells were maintained at 37°C in a 5% CO2 incubator with DMEM (Invitrogen, 85Fisher Scientific UK Ltd, Loughborough, UK) supplemented with 10% foetal bovine 86 serum, 100 U/ml penicillin, 100 µg/ml streptomycin. Experimental treatment of cells 87 included incubation in medium supplemented with 200 µg/ml cycloheximide (Sigma, 88 Gillingham, UK) and/or 5 µg/ml actinomycin D (Cayman Chemical Cambridge 89Bioscience Ltd, Cambridge, UK). 90 91 DNA constructs 92Generation of some constructs relied on gene synthesis (Invitrogen, Fisher Scientific 93 UK Ltd, Loughborough, UK). In instances where this was the case, sequences ordered 94 were codon optimized for mammalian expression and then manually adjusted to 95 minimize the existence of unwanted splice donor and splice acceptor sites using the 96 online programmes HSF3[25] and the NetGene2 server [26]. The initial plasmid 97 expressing GFP, VSV-tagged 2A and HA-tagged 3C from HRV16 was generated by 98 cloning a single synthetic DNA fragment into an in-house dual promoter plasmid via 99EcoRI and SalI restriction sites to generate pCIPEP-A16 TAG (+/+) (see Fig. S1 for 100 sequence). Synthesized DNAs encoding for HRV-B4 and HRV-C2 proteases (Table 101 S1) were exchanged with their respective counterparts in pCIPEP-A16 TAG (+/+) using 102ClaI and SbfI (2A) and BsiWI and SalI (3C) restriction sites. Inactivation of the 103 proteases involved a two-step PCR approach, which introduced a Cys>Ala mutation 104 in the active site of both 2A and 3C. A similar PCR-based strategy was used to 105 generate constructs expressing tag-free proteases and a 3C protease with a c-Myc 106 nuclear localization signal (PAAKRVKLD) fused directly to its C-terminus. Primer 107 sequences are available on request. 108The synthetic DNA encoding for the Thioredoxin-Nanoluciferase(Nluc)PEST 109 fusion protein linked by a peptide derived from translating the hepatitis delta virus 110 (HdV) ribozyme sequence (HdV WT Nluc; Fig. S2) was initially cloned into pCDNA3.

Core medical training was replaced by Internal Medicine Training (IMT) in August 2019. One of the more notable changes was the introduction of a 3 We created an in-person teaching day, entitled ‘Step up’, which consisted of four 20-minute, IM3 specific, high-fidelity scenario-based simulations. In each scenario, a computerised full-body manikin (SimMan Essential) was used. The manikin could be programmed to provide physiological response to scenario participants’ actions. The participants were each tasked to assess an acutely deteriorating patient (the manikin), whilst managing a demanding bleep to add realistic distractions and human factor issues to the scenario. Each participant completed one scenario and watched their peers complete the others from a separate room. Following this, participants were debriefed by qualified member of staff and then taught by a speciality registrar or consultant. Feedback forms were collected. The participants were tasked to rate the simulation day, using a nominal Likert scale from 1 to 10, for usefulness, relevance, clarity, and overall quality. 1 represented strong disagreement and 10 strong agreements with a particular domain. Across 4 full days, we had 11 IM3 doctors take part in the ‘step up’ scenario-based simulations. 9 of the 11 participants completed feedback forms. Scores ranged from 8 to 10. Median scores for all domains were 10. We have demonstrated that ‘step up’ scenarios are a useful and relevant aid for those transitioning into IM3. Further research utilising comparative data will provide more meaningful conclusions. We will be repeating the simulation programme for the next cohort. We will have baseline, one and three-month follow up questionnaires to assess these scenarios further. 1. Buist N, Webster CS. Simulation training to improve the ability of first-year doctors to assess and manage deteriorating patients: a systematic review and meta-analysis. Medical Science Educator. 2019;29(3):749–761.

Human rhinoviruses (HRVs) express 2 cysteine proteases, 2A and 3C, that are responsible for viral polyprotein processing. Both proteases also suppress host gene expression by inhibiting mRNA transcription, nuclear export and cap-dependent translation. However, the relative contribution that each makes in achieving this goal remains unclear. In this study, we have compared both the combined and individual ability of the two proteases to shut down cellular gene expression using a novel dynamic reporter system. Our findings show that 2A inhibits host gene expression much more rapidly than 3C. By comparing the activities of a representative set of proteases from the three different HRV species, we also find variation in the speed at which host gene expression is suppressed. Our work highlights the key role that 2A plays in early suppression of the infected host cell response and shows that this can be influenced by natural variation in the activity of this enzyme.