Inhibitors of the tyrosine kinase Btk have been proposed as novel antiplatelet agents. In this study we show that low concentrations of the Btk inhibitor ibrutinib block CLEC-2-mediated activation and tyrosine phosphorylation including Syk and PLCγ2 in human platelets. Activation is also blocked in patients with X-linked agammaglobulinemia (XLA) caused by a deficiency or absence of Btk. In contrast, the response to GPVI is delayed in the presence of low concentrations of ibrutinib or in patients with XLA, and tyrosine phosphorylation of Syk is preserved. A similar set of results is seen with the second-generation inhibitor, acalabrutinib. The differential effect of Btk inhibition in CLEC-2 relative to GPVI signalling is explained by the positive feedback role involving Btk itself, as well as ADP and thromboxane A2 mediated activation of P2Y12 and TP receptors, respectively. This feedback role is not seen in mouse platelets and, consistent with this, CLEC-2-mediated activation is blocked by high but not by low concentrations of ibrutinib. Nevertheless, thrombosis was absent in 8 out of 13 mice treated with ibrutinib. These results show that Btk inhibitors selectively block activation of human platelets by CLEC-2 relative to GPVI suggesting that they can be used at 'low dose' in patients to target CLEC-2 in thrombo-inflammatory disease.

White-nose syndrome (WNS) has rendered four of Ontario’s species endangered, while leaving the other four species relatively unaffected. The causes and extent of the declines have been widely studied. The influence on remaining bat species has not. Comparing acoustic data recorded ∼10 years apart, we evaluated how species in southeastern Ontario, Canada, use different foraging habitats pre- and post-WNS detection. We observed activity declines in now-endangered species over open fields (small-footed myotis, Myotis leibii (Audubon and Bachman, 1842); little brown bat, Myotis lucifugus (Le Conte, 1831); northern myotis, Myotis septentrionalis (Trouessart, 1897); tricolored bat, Perimyotis subflavus (F. Cuvier, 1832)) and speculate that the reduction of the once most common species (M. lucifugus) may have resulted in other species searching for prey in habitat once dominated by M. lucifugus. That is, these changes may have allowed greater presence in open field and clutter or edge environments by the big brown bat (Eptesicus fuscus (Palisot de Beauvois, 1796)) and three migratory species (silver-haired bat, Lasionycteris noctivagans (Le Conte, 1831); red bat, Lasiurus borealis (Müller, 1776); hoary bat, Lasiurus cinereus (Palisot de Beauvois, 1796)). However, our results also suggest that (i) while the decline of most resident bat species due to WNS may have relaxed competition for relatively unaffected species in some, but not all habitats, that (ii) sensory and biomechanical constraints may limit prey exploitation by these less-affected bat species in these habitats.

Understanding how wildlife respond to ever-encroaching urbanization is of great concern. Bats are the second-most speciose mammalian order and while many appear to be urban adapted, we currently have a limited understanding of their demography and habitat use within urban environments. Using a combination of captures to obtain demographic data, radio-telemetry to examine foraging and roosting behaviour, and data on diet and prey availability, we examined how big brown bats (Eptesicus fuscus), a synurbic species, use an urban green space (High Park) in Canada’s largest city centre, Toronto. We found that adult males outnumbered adult females more than two to one and that males were found throughout the park, while females were concentrated in an area with greater access to water, but lower prey availability. We also found that bats of both sexes were in poorer body condition than reported for other non-urban areas, including a site within southern Ontario. Our data suggest that High Park may not provide adequate resources for reproductive females as they were never found roosting in the park and beetles, their preferred prey, were limited. Although previous studies suggest urban green spaces may offer refuge to bats, most have not considered sex-specific responses to urbanization as they have largely been based on acoustic surveys. Our study therefore highlights the importance of considering demographic differences in response to urbanization to better inform urban management plans and green space development.