http://bura.brunel.ac.uk/handle/2438/8614 2026-06-30T16:16:07Z http://bura.brunel.ac.uk/handle/2438/33468 Title: Visualizing the interplay of Cas1–Cas2 with DNA replication-repair that creates CRISPR–Cas immunity Authors: Hashemloo, MA; Killelea, T; Mamić, T; Ireland, TH; Lou-Hing, A; Kemm, F; Dimude, JU; Žagar, M; Ivančić-Baće, I; Rudolph, CJ; Bolt, EL Abstract: Prokaryotic CRISPR–Cas systems rely on the Cas1–Cas2 protein complex to capture new DNA from mobile genetic elements (MGEs), to form immunological memory that defends against the MGEs. However, the mechanisms by which Cas1–Cas2 locates suitable DNA substrates inside cells remain unclear, limiting our understanding of how CRISPR–Cas immunity arises de novo. We directly visualized functional, DNA-bound Cas1–Cas2 complexes in bacteria, revealing the processes that license Cas1–Cas2 to capture DNA. Visible DNA-bound Cas1–Cas2 complexes formed only when replisomes are actively advancing, accumulating at post-replicative DNA gaps behind replication forks—structures arising during normal genome duplication, which are normally repaired by homologous recombination. Replication stress, which increases replicative DNA gap frequency, enhanced visible Cas1–Cas2 DNA binding. DNA capture by Cas1–Cas2 was strongly stimulated in cells lacking the RecFOR complex, which normally directs DNA gaps to repair. The RecBCD recombination initiator complex was essential for DNA capture by Cas1–Cas2 in these cells. The findings support a model in which naïve CRISPR–Cas adaptation is licensed by abundant replication-dependent DNA repair intermediates, prior to their repair by recombination. This identifies the mechanism co-ordinating Cas1–Cas2 with essential DNA replication and repair processes that all cells need, including when they are hijacked to replicate parasitic MGEs. Description: Data availability: The data referred to in this work have been deposited in Figshare (https://figshare.com) with digital object identifier: 10.6084/m9.figshare.30813251; Supplementary data are available online at: https://academic.oup.com/nar/article/54/11/gkag564/8703704?login=true#565026736 .; M. Amin Hashemloo, Tom Killelea and Tomislav Mamić should be regarded as joint first authors. 2026-06-08T00:00:00Z http://bura.brunel.ac.uk/handle/2438/33460 Title: Effects of aging on recognition and dominance perception in laughter Authors: Szameitat, DP; Wildgruber D; Szameitat, AJ Abstract: Introduction: Aging is associated with reduced accuracy in recognizing others’ emotions, an ability that is important for maintaining social connectedness in later life. Laughter is a social signal with multiple functions, as it can facilitate social bonding but also convey negative social meanings, for example when directed at someone. In previous research we have shown that younger adults are able to classify spontaneously emitted joyful, schadenfreude, and tickling laughter above chance level, and that these laughter sounds differ according to the perceived dominance. Given evidence that affect recognition generally declines with age, the present study examined whether comparable age effects emerge in the perception of laughter. Methods: 64 younger adults (mean 25 years, 18–33 years) and 30 older adults (mean age 60 years, 50–77 years) evaluated 117 spontaneously emitted laughter sounds according to the laughter type, i.e., joyful, Schadenfreude, and tickling laughter and according to the perceived sender’s dominance. Results: Results showed that both age groups classified laughter above chance level. Younger adults showed higher classification rates than older adults for all laughter types, with the largest age effect for Schadenfreude laughter. The dominance ratings showed an age effect only for Schadenfreude, where older adults rated Schadenfreude laughter less dominant than younger adults. Discussion: Pronounced differences in Schadenfreude perception might be ascribed to difficulties of older adults in perceiving non-literal messages or to cultural differences between age groups. Description: Data availability statement: The original contributions presented in the study are publicly available. This data can be found here: Figshare, https://doi.org/10.17633/rd.brunel.32521458. 2026-06-16T00:00:00Z http://bura.brunel.ac.uk/handle/2438/33453 Title: Toads on Roads: Summary 2025 Authors: Sumpter, JP Abstract: 2025 Data Collection: Data collection is now complete for another year with results from 280 patrols processed. 2025-12-01T00:00:00Z http://bura.brunel.ac.uk/handle/2438/33451 Title: Embryotoxicity of pyrethroid insecticides and their mixtures in a human induced pluripotent stem cell-based model in vitro Authors: Ma, Y; Gutierrez, CT; Scholze, M; Valente, MJ; Vinggaard, AM Abstract: Pyrethroid insecticides are used worldwide, yet their potential effects on early human development remain poorly understood. We applied the PluriLum assay, a human induced pluripotent stem cell-based 3D in vitro model, to evaluate the embryotoxicity of three commonly used pyrethroids (α-cypermethrin, deltamethrin, and etofenprox), and their shared metabolite 3-phenoxybenzoic acid (3-PBA). Embryoid bodies (EBs) were repeatedly exposed to these compounds throughout their differentiation into cardiomyocytes, and effects on cardiomyocyte beating, expression of the early cardiac marker NKX2.5, as well as their impact at a transcriptional level was assessed. The three pyrethroids were tested in combination at two different mixture compositions to investigate their potential for additivity. All three pyrethroids impaired cardiomyocyte differentiation at micromolar levels, with the following potency ranking: α-cypermethrin > etofenprox > deltamethrin. At higher concentrations, α-cypermethrin also reduced cardiomyocyte contractility. In contrast, 3-PBA showed no significant effects on neither differentiation nor contractile function. The pyrethroid mixtures followed the principle of concentration addition. An exploratory transcriptomic analysis revealed that α-cypermethrin and deltamethrin significantly altered the EBs gene expression profiles, affecting pathways related to ion channel activity, receptor signalling, and extracellular matrix organization. These findings suggest that exposure to pyrethroids may interfere with early human cardiac development through effects on multiple molecular targets. This study also highlights the value of the PluriLum assay as a human-relevant platform for assessing the embryotoxic potential of environmental chemicals. Description: Data availability: The datasets generated during and/or analysed during the current study are available from the corresponding author on reasonable request.; Supplementary Information is available online at: https://link.springer.com/article/10.1007/s00204-026-04448-2#Sec211 . 2026-05-20T00:00:00Z