Tn-Seq reveals hidden complexity in the utilization of host-derived glutathione in \u3cem\u3eFrancisella tularensis\u3c/em\u3e

Host-derived glutathione (GSH) is an essential source of cysteine for the intracellular pathogen Francisella tularensis. In a comprehensive transposon insertion sequencing screen, we identified several F. tularensis genes that play central and previously unappreciated roles in the utilization of GSH during the growth of the bacterium in macrophages. We show that one of these, a gene we named dptA, encodes a proton-dependent oligopeptide transporter that enables growth of the organism on the dipeptide Cys-Gly, a key breakdown product of GSH generated by the enzyme γ-glutamyltranspeptidase (GGT). Although GGT was thought to be the principal enzyme involved in GSH breakdown in F. tularensis, our screen identified a second enzyme, referred to as ChaC, that is also involved in the utilization of exogenous GSH. However, unlike GGT and DptA, we show that the importance of ChaC in supporting intramacrophage growth extends beyond cysteine acquisition. Taken together, our findings provide a compendium of F. tularensis genes required for intracellular growth and identify new players in the metabolism of GSH that could be attractive targets for therapeutic intervention

An exploration of the relationship between educational background and the coaching behaviours and practice activities of professional youth soccer coaches

This paper is closed access until 12 December 2019.Background and purpose: Despite the proliferation in recent years of higher education establishments offering tertiary-level study in the field of sports coaching, there is a lack of research into the impact of such courses on coaching practice. The behaviours employed and activities used by coaches during practice sessions is an area where one might expect to see such impact, indeed certain studies have tentatively noted the educational qualifications of coaches and suggested that this may play a role in the application of behaviours more aligned with player learning. The purpose of this study was therefore to compare youth soccer coaches with and without tertiary-level qualifications, examining their coaching behaviours and practice activities. Method: The participants were 10 male professional youth soccer coaches aged 24–55 with an average of 13 years coaching experience. Five of the coaches had completed undergraduate degree courses related to sport coaching. All of the coaches worked with players aged under 9 to under 18 in the youth academy of an English professional soccer club. Systematic observation of coach behaviour and practice activities was carried out using the Coach Analysis and Intervention System (Cushion et al. 2012), while follow-up interviews were used to elicit the coaches’ perceptions of, and rationale for, their behaviour. Findings: The observation data showed that graduate coaches used significantly more divergent questioning than non-graduate coaches, while the interview data revealed a general trend for graduate coaches to show greater self-awareness of behaviours and changes in behaviour between practice types. Graduate coaches also provided more comprehensive rationales, for example, seeing silence as a means of facilitating player decision-making as well as for observation. In contrast to previous research, sessions featured a higher proportion of playing form than training form activities and at over 20% of session duration, the ‘other’ practice state was a prominent feature of contact time with players. While some coaches saw ‘other’ as wasted time, graduate coaches identified this as an opportunity for group discussion and social interaction. The study adds to existing data about coach behaviours and practice activities, providing evidence that education background may indeed influence coaching practice