Elucidation of a bacteriophytochrome-regulated signal transduction pathway in Pseudomonas syringae that contributes to leaf colonization, virulence, and swarming motility

Plant-associated bacteria encounter a range of stressful environmental conditions when colonizing leaf surfaces. To adapt to these harsh conditions bacteria sense and respond to environmental signals. Within the last two decades, photoreceptors that respond to specific wavelengths of light through associated chromophores have been discovered with increasing frequency in non-photosynthetic bacteria, including those associated with plants. Their presence suggests that fluctuations in light may serve as a cue to regulate bacterial adaptations. The foliar plant pathogen Pseudomonas syringae is unusual among heterotrophic bacteria because it encodes three photoreceptors, two red- and/or far-red light-sensing bacteriophytochromes and a blue light-sensing LOV protein. Here we evaluated the physiological roles of these photoreceptors, their mechanisms of regulation, and their impacts on plant colonization. This work provides the first evidence in bacteria for an integrated signaling network composed of both a LOV protein and a phytochrome, and shows that the bacteriophytochrome, BphP1, and LOV control swarming motility. BphP1 represses swarming motility in response to red and far-red light, whereas LOV attenuates BphP1-mediated repression. Moreover, this is the first bacteriophytochrome shown to have blue-light sensing capabilities, and these occur independently of red-light sensing. Furthermore, this work identifies a role for a bacteriophytochrome in plant colonization for the first time and demonstrates that this bacteriophytochrome, BphP1, promotes survival during the initial stages of leaf colonization and negatively regulates colonization later on. BphP1-mediated regulation of swarming motility is associated with the ability of P. syringae to move from soil to seeds and contributes to lesion development. This work further elucidates the mechanism of BphP1-mediated regulation and demonstrates that BphP1 and a regulator we designate Lsr repress swarming motility in response to red light by controlling the transition from a sessile to motile lifestyle. Additionally, an acyl-homoserine lactone molecule functions as a downstream component in this signal transduction pathway. This work also provides evidence for light-mediated regulation of the type IV pilus and demonstrates for the first time a role for type IV pili in the swarming motility of P. syringae pv. syringae. Furthermore, the global regulator AlgU is shown to negatively regulate swarming independent of alginate production, which itself enhances swarming motility. Finally, the work documents an unusual interaction between P. syringae colonies that manifests as induced movement away from colonies producing the repellent 3-(3-hydroxyalkanoyloxy) alkanoic acid by strain derivatives that otherwise appear non-motile

Chiral Ring Generating Functions & Branches of Moduli Space

34 pages, 7 figures34 pages, 7 figures34 pages, 7 figuresWe consider the worldvolume theory of N D3-branes transverse to various non-compact Calabi-Yau spaces, and describe subtleties in the counting of chiral primary operators in such theories due to the presence of multiple branches of moduli space. Extra branches, beyond those directly related to the transverse geometry, result in additional terms in the generating functions for single- and multi-trace operators. Ideals in the N=1 chiral ring correspond to various branches and, in the large N limit, the operator counting reveals a product of Fock spaces, including the Fock space of bosons on the space transverse to the branes

Investigation into the relationship between adolescents’ perceived and actual fundamental movement skills and physical activity

Abstract Objectives: To explore the relationship between fundamental movement skill (FMS) competence, perceived FMS competence and physical activity (PA) in adolescents. Methods: The Test of Gross Motor Development (TGMD), the TGMD-2 and the Victorian Skills manual were used to assess FMS competence (locomotor, object control and stability). The Physical Self Confidence scale was used to assess perceived FMS competence (locomotor, object control and stability). Moderate to vigorous intensity PA (MVPA) was measured via accelerometry. Multi-level modelling analyses was used to examine: (i) actual FMS as the predictor and perceived FMS as the outcome, (ii) perceived FMS as the predictor and MVPA as the outcome, and (iii) actual FMS as the predictor and MVPA as the outcome. All analyses were completed for each subtest of FMS (locomotor, object control and stability). Results: A total of 584 adolescents (boys n = 278) aged 12.82 – 15.25 years (M 13.78, SD .42) participated in this study. Actual stability was associated with perceived stability (p<.01) and MVPA (p<.05) in boys. This was not found true for girls, however actual locomotor skills were associated with MVPA (p≤.05). Boys scored significantly higher than girls for FMS proficiency, perceived FMS and MVPA (p<.05). Discussion: Gender differences may exist due to cultural gender differences in sport participation norms. Considering the magnitude of physical and psychological changes occurring during adolescence, it is recommended to track young people over time to better understand the relationship between perceived and actual FMS, as well as PA participation. Keywords: physical activity, motor competence, adolescent

The Youth-Physical Activity Towards Health (Y-PATH) intervention: Results of a 24 month cluster randomised controlled trial

Low levels of physical activity in youth are an issue internationally, with the age related decline in levels over the adolescent period of particular concern. This study evaluated a multi-component school-based intervention (Y-PATH: Youth-Physical Activity Towards Health), focused on halting the age-related decline in physical activity of youth in early adolescence. A cluster randomized controlled trial in 20 post primary schools (10 control, 10 intervention) was conducted. Data were collected from all 20 schools at baseline (2013), and 12 months (2014), and from 10 of these schools (5 intervention) at 24 months (2015). The setting was mixed gender post primary schools residing in the greater area of Dublin, Ireland. Principals from each school were asked to nominate one first year class group attending their school in September 2013 to participate in the study (N = 564). Intervention schools implemented the Y-PATH whole school intervention, comprising teacher component, parent component, and PE component; while control schools continued with usual care. The main outcome measure was accelerometer derived average minutes of daily moderate to vigorous physical activity (MVPA). Data were analysed from October 2015 -November 2017. At baseline 490 participants were assessed (mean age 12.78y ± .42). Results of the multilevel regression analysis confirmed that there was a significant time intervention effect, and this was predominantly contributed by the difference between control and intervention groups within females. Findings support the case for national dissemination of the Y-PATH intervention so that the knowledge learned can be translated to routine practice in schools

Reducing delays in the diagnosis and treatment of muscle-invasive bladder cancer using simulation modelling

This is the author accepted manuscript. The final version is available from SAGE Publications via the DOI in this record Objective: To develop a simulation model to identify key bottlenecks in the bladder cancer pathway at Royal Cornwall Hospital and predict the impact of potential changes to reduce these delays. Materials and methods: The diagnosis and treatment of muscle-invasive bladder cancer can suffer numerous delays, which can significantly affect patient outcomes. We developed a discrete event computer simulation model of the flow of patients through the bladder cancer pathway at the hospital, using anonymised patient records from 2014 and 2015. The changes tested in the model were for patients suspected to have muscle-invasive disease on flexible cystoscopy. Those patients were ‘fast-tracked’ to receive their transurethral resection of bladder tumour (TURBT) treatment using operating slots kept free for these patients. A staging computed tomography scan was booked in the haematuria clinic. Pathology requests were marked as 48 hour turnaround. The nurse specialist would then speak to the patient whilst they were on the ward following their TURBT to give information about their ongoing treatment and provide support. Results: The model predicted that if the changes were implemented, delays in the system could be reduced by around 5 weeks. The changes were implemented, and analysis of 3 months of the data post-implementation shows that the average time in the system was reduced by 5 weeks. The environment created by the changes in the pathway improved referral to treatment times in both muscle-invasive and non-muscle-invasive groups. Conclusion: The simulation model proved an invaluable tool for facilitating the implementation of changes. Simple changes to the pathway led to significant reductions in delays for bladder cancer patients at Royal Cornwall Hospital. Level of evidence: Not applicable for this cohort study.National Institute for Health Research (NIHR

Gambling adverts in live TV coverage of the Qatar 2022 FIFA Men’s World Cup

Background: Gambling marketing is ubiquitous in UK football and, despite gambling industry self-regulation such as the whistle-to-whistle ban, remains prominent in live TV coverage. Major international tournaments do not usually feature gambling pitch-side advertising and shirt sponsorship, increasing the importance of TV adverts during these high-profile competitions. The present study examined the prevalence and features of gambling adverts shown during the commercial broadcaster ITV’s live coverage of games in the 2022 Qatar World Cup. Method: Each match shown live on ITV was recorded. For each gambling advert, the timing of the advert (pre-/during-/post-match), the advert category (financial inducements/live odds/safer gambling/brand awareness) and safer gambling messaging were recorded. Results: Over the 30 matches analyzed, there were 156 adverts directly for gambling brands or products (M = 5.2, range 3–8), featuring adverts from eight different operators. The majority of adverts were shown pre-match (80.8%). Financial inducements were most commonly advertised (42.3%), followed by brand awareness adverts (26.9%). The safer gambling message ‘take time to think’ was shown in 70.5% of adverts. Adverts for lottery products did not feature any safer gambling messages. Conclusions: Multiple gambling adverts were shown during each match of the 2022 Qatar World Cup, especially so pre-match. Pre-match adverts predominantly encourage viewers to gamble promptly, through financial inducements and boosted live odds. Any potential further legislation could therefore consider either further restrictions based on the entire broadcast, or by enforcing the use of specific safer gambling messages.</p

Structure-based discovery of glycomimetic FmlH ligands as inhibitors of bacterial adhesion during urinary tract infection

Significance The emergence of multidrug-resistant bacteria, including uropathogenic Escherichia coli (UPEC), makes the development of targeted antivirulence therapeutics a critical focus of research. During urinary tract infections (UTIs), UPEC uses chaperone–usher pathway pili tipped with an array of adhesins that recognize distinct receptors with sterochemical specificity to facilitate persistence in various tissues and habitats. We used an interdisciplinary approach driven by structural biology and synthetic glycoside chemistry to design and optimize glycomimetic inhibitors of the UPEC adhesin FmlH. These inhibitors competitively blocked FmlH in vitro, in in vivo mouse UTI models, and in ex vivo healthy human kidney tissue. This work demonstrates the utility of structure-driven drug design in the effort to develop antivirulence therapeutic compounds. </jats:p

Carbohydrate restriction and dietary cholesterol modulate the expression of HMG-CoA reductase and the LDL receptor in mononuclear cells from adult men

The liver is responsible for controlling cholesterol homeostasis in the body. HMG-CoA reductase and the LDL receptor (LDL-r) are involved in this regulation and are also ubiquitously expressed in all major tissues. We have previously shown in guinea pigs that there is a correlation in gene expression of HMG-CoA reductase and the LDL-r between liver and mononuclear cells. The present study evaluated human mononuclear cells as a surrogate for hepatic expression of these genes. The purpose was to evaluate the effect of dietary carbohydrate restriction with low and high cholesterol content on HMG-CoA reductase and LDL-r mRNA expression in mononuclear cells. All subjects were counseled to consume a carbohydrate restricted diet with 10–15% energy from carbohydrate, 30–35% energy from protein and 55–60% energy from fat. Subjects were randomly assigned to either EGG (640 mg/d additional dietary cholesterol) or SUB groups [equivalent amount of egg substitute (0 dietary cholesterol contributions) per day] for 12 weeks. At the end of the intervention, there were no changes in plasma total or LDL cholesterol (LDL-C) compared to baseline (P > 0.10) or differences in plasma total or LDL-C between groups. The mRNA abundance for HMG-CoA reductase and LDL-r were measured in mononuclear cells using real time PCR. The EGG group showed a significant decrease in HMG-CoA reductase mRNA (1.98 ± 1.26 to 1.32 ± 0.92 arbitrary units P < 0.05) while an increase was observed for the SUB group (1.13 ± 0.52 to 1.69 ± 1.61 arbitrary units P < 0.05). Additionally, the LDL-r mRNA abundance was decreased in the EGG group (1.72 ± 0.69 to 1.24 ± 0.55 arbitrary units P < 0.05) and significantly increased in the SUB group (1.00 ± 0.60 to 1.67 ± 1.94 arbitrary units P < 0.05). The findings indicate that dietary cholesterol during a weight loss intervention alters the expression of genes regulating cholesterol homeostasis

Differences in level of confidence in diabetes care between different groups of trainees: the TOPDOC diabetes study

Background There is an increasing prevalence of diabetes. Doctors in training, irrespective of specialty, will have patients with diabetes under their care. The aim of this further evaluation of the TOPDOC Diabetes Study data was to identify if there was any variation in confidence in managing diabetes depending on the geographical location of trainees and career aspirations. Methods An online national survey using a pre-validated questionnaire was administered to trainee doctors. A 4-point confidence rating scale was used to rate confidence in managing aspects of diabetes care and a 6-point scale used to quantify how often trainees would contribute to the management of patients with diabetes. Responses were grouped depending on which UK country trainees were based and their intended career choice. Results Trainees in Northern Ireland reported being less confident in IGT diagnosis, use of IV insulin and peri-operative management and were less likely to adjust oral treatment, contact specialist, educate lifestyle, and optimise treatment. Trainees in Scotland were less likely to contact a specialist, but more likely to educate on lifestyle, change insulin, and offer follow-up advice. In Northern Ireland, Undergraduate (UG) and Postgraduate (PG) training in diagnosis was felt less adequate, PG training in emergencies less adequate, and reporting of need for further training higher. Trainees in Wales felt UG training to be inadequate. In Scotland more trainees felt UG training in diagnosis and optimising treatment was inadequate. Physicians were more likely to report confidence in managing patients with diabetes and to engage in different aspects of diabetes care. Aspiring physicians were less likely to feel the need for more training in diabetes care; however a clear majority still felt they needed more training in all aspects of care. Conclusions Doctors in training have poor confidence levels dealing with diabetes related care issues. Although there is variability between different groups of trainees according to geographical location and career aspirations, this is a UK wide issue. There should be a UK wide standardised approach to improving training for junior doctors in diabetes care with local training guided by specific needs.</p