A global genomic approach uncovers novel components for twitching motility-mediated biofilm expansion in Pseudomonas aeruginosa.

Pseudomonas aeruginosa is an extremely successful pathogen able to cause both acute and chronic infections in a range of hosts, utilizing a diverse arsenal of cell-associated and secreted virulence factors. A major cell-associated virulence factor, the Type IV pilus (T4P), is required for epithelial cell adherence and mediates a form of surface translocation termed twitching motility, which is necessary to establish a mature biofilm and actively expand these biofilms. P. aeruginosa twitching motility-mediated biofilm expansion is a coordinated, multicellular behaviour, allowing cells to rapidly colonize surfaces, including implanted medical devices. Although at least 44 proteins are known to be involved in the biogenesis, assembly and regulation of the T4P, with additional regulatory components and pathways implicated, it is unclear how these components and pathways interact to control these processes. In the current study, we used a global genomics-based random-mutagenesis technique, transposon directed insertion-site sequencing (TraDIS), coupled with a physical segregation approach, to identify all genes implicated in twitching motility-mediated biofilm expansion in P. aeruginosa. Our approach allowed identification of both known and novel genes, providing new insight into the complex molecular network that regulates this process in P. aeruginosa. Additionally, our data suggest that the flagellum-associated gene products have a differential effect on twitching motility, based on whether components are intra- or extracellular. Overall the success of our TraDIS approach supports the use of this global genomic technique for investigating virulence genes in bacterial pathogens

Physiological Characteristics of Female Soccer Players and Health and Performance Considerations: A Narrative Review

From Springer Nature via Jisc Publications RouterHistory: accepted 2021-03-22, registration 2021-03-22, online 2021-04-12, pub-electronic 2021-04-12, pub-print 2021-07Publication status: PublishedAbstract: Female soccer has seen a substantial rise in participation, as well as increased financial support from governing bodies over the last decade. Thus, there is an onus on researchers and medical departments to develop a better understanding of the physical characteristics and demands, and the health and performance needs of female soccer players. In this review, we discuss the current research, as well as the knowledge gaps, of six major topics: physical demands, talent identification, body composition, injury risk and prevention, health and nutrition. Data on female talent identification are scarce, and future studies need to elucidate the influence of relative age and maturation selection across age groups. Regarding the physical demands, more research is needed on the pattern of high-intensity sprinting during matches and the contribution of soccer-specific movements. Injuries are not uncommon in female soccer players, but targeting intrinsically modifiable factors with injury prevention programmes can reduce injury rates. The anthropometric and physical characteristics of female players are heterogeneous and setting specific targets should be discouraged in youth and sub-elite players. Menstrual cycle phase may influence performance and injury risk; however, there are few studies in soccer players. Nutrition plays a critical role in health and performance and ensuring adequate energy intake remains a priority. Despite recent progress, there is considerably less research in female than male soccer players. Many gaps in our understanding of how best to develop and manage the health and performance of female soccer players remain

The stigmatisation of source isolation: a literature review

Background: Isolation precautions in patients with multi-drug-resistant bacteria and other communicable infectious agents can be associated with adverse effects. Patients’ perspectives of isolation suggest that the imposed environment and procedures create barriers to their physical, social and emotional needs.Aims: The purpose of this paper is to review the literature to uncover any reliable evidence supporting the assertion that stigma is a significant characteristic of the experience of source isolation in healthcare settings.Methods: The methodological framework of Arksey and O’Malley was applied to this review. A total of 14 papers identified from 189 abstracts screened were included in the review.Results: The research reviewed suggests a clear association between stigmatisation and isolation in which stigma does have a direct negative effect on patients placed in hospital isolation. None of the studies found evidence to the contrary.Conclusions: The implications of this literature review for policy-makers and healthcare professionals suggest that when isolation or other forms of constraint are implemented and in use, patients must be provided with strengthened forms of support, including social and emotional support, and given access to healthcare of optimal quality to prevent the associated adverse effects of isolation as much as possibl

Spliced Leader Trapping Reveals Widespread Alternative Splicing Patterns in the Highly Dynamic Transcriptome of Trypanosoma brucei

Trans-splicing of leader sequences onto the 5′ends of mRNAs is a widespread phenomenon in protozoa, nematodes and some chordates. Using parallel sequencing we have developed a method to simultaneously map 5′splice sites and analyze the corresponding gene expression profile, that we term spliced leader trapping (SLT). The method can be applied to any organism with a sequenced genome and trans-splicing of a conserved leader sequence. We analyzed the expression profiles and splicing patterns of bloodstream and insect forms of the parasite Trypanosoma brucei. We detected the 5′ splice sites of 85% of the annotated protein-coding genes and, contrary to previous reports, found up to 40% of transcripts to be differentially expressed. Furthermore, we discovered more than 2500 alternative splicing events, many of which appear to be stage-regulated. Based on our findings we hypothesize that alternatively spliced transcripts present a new means of regulating gene expression and could potentially contribute to protein diversity in the parasite. The entire dataset can be accessed online at TriTrypDB or through: http://splicer.unibe.ch/

Regulation of Trypanosoma brucei Total and Polysomal mRNA during Development within Its Mammalian Host

This work was supported by a Wellcome Trust Programme grant to KM and by a Wellcome Trust Strategic award to the Centre for Immunity, Infection and Evolution at the University of Edinburgh. SM was supported by a studentship from the Medical Research Council, UK.The gene expression of Trypanosoma brucei has been examined extensively in the blood of mammalian hosts and in forms found in the midgut of its arthropod vector, the tsetse fly. However, trypanosomes also undergo development within the mammalian bloodstream as they progress from morphologically 'slender forms' to transmissible 'stumpy forms' through morphological intermediates. This transition is temporally progressive within the first wave of parasitaemia such that gene expression can be monitored in relatively pure slender and stumpy populations as well as during the progression between these extremes. The development also represents the progression of cells from translationally active forms adapted for proliferation in the host to translationally quiescent forms, adapted for transmission. We have used metabolic labelling to quantitate translational activity in slender forms, stumpy forms and in forms undergoing early differentiation to procyclic forms in vitro. Thereafter we have examined the cohort of total mRNAs that are enriched throughout development in the mammalian bloodstream (slender, intermediate and stumpy forms), irrespective of strain, revealing those that exhibit consistent developmental regulation rather than sample specific changes. Transcripts that cosediment with polysomes in stumpy forms and slender forms have also been enriched to identify transcripts that escape translational repression prior to transmission. Combined, the expression and polysomal association of transcripts as trypanosomes undergo development in the mammalian bloodstream have been defined, providing a resource for trypanosome researchers. This facilitates the identification of those that undergo developmental regulation in the bloodstream and therefore those likely to have a role in the survival and capacity for transmission of stumpy forms.Publisher PDFPeer reviewe

Genomic reconstruction of the SARS-CoV-2 epidemic in England.

The evolution of the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) virus leads to new variants that warrant timely epidemiological characterization. Here we use the dense genomic surveillance data generated by the COVID-19 Genomics UK Consortium to reconstruct the dynamics of 71 different lineages in each of 315 English local authorities between September 2020 and June 2021. This analysis reveals a series of subepidemics that peaked in early autumn 2020, followed by a jump in transmissibility of the B.1.1.7/Alpha lineage. The Alpha variant grew when other lineages declined during the second national lockdown and regionally tiered restrictions between November and December 2020. A third more stringent national lockdown suppressed the Alpha variant and eliminated nearly all other lineages in early 2021. Yet a series of variants (most of which contained the spike E484K mutation) defied these trends and persisted at moderately increasing proportions. However, by accounting for sustained introductions, we found that the transmissibility of these variants is unlikely to have exceeded the transmissibility of the Alpha variant. Finally, B.1.617.2/Delta was repeatedly introduced in England and grew rapidly in early summer 2021, constituting approximately 98% of sampled SARS-CoV-2 genomes on 26 June 2021

Carbon Monoxide Poisoning: Saving Lives, Advancing Treatment

Carbon monoxide (CO) poisoning is a serious public health issue. In England and Wales alone, every year some 4,000 attendances to emergency departments (EDs) are the result of accidental CO poisoning. Statistics show that CO kills more than 30 people a year and leads to around 200 hospital admissions, but these figures are likely to be a gross underestimate. Consequently, treating accidental CO poisoning may actually be costing much more than the estimated £178 million per annum. Healthcare professionals have a vital role to play in preventing, diagnosing and treating patients exposed to CO. However, these professionals face a number of barriers to action: gaps in knowledge, limited awareness, and a lack of co-ordination within and between the healthcare sectors. These barriers need to be removed if we are to reduce significantly the number of accidental deaths and unnecessary injuries caused by CO poisoning, and to improve patient management and recovery. This report has been prepared by members of COMed, the healthcare professionals’ sub-group of the APPCOG Stakeholder Forum. It presents a number of hard-hitting essays that review current knowledge and practice on the diagnosis and management of CO poisoning in the healthcare system. It identifies gaps in knowledge and practice, and makes recommendations to close those gaps so that diagnosis, patient management and recovery can be improved. The findings presented in this report led members of the sub-group to conclude that: A lack of awareness amongst healthcare professionals of CO poisoning as a cause of illness is very likely to be impacting adversely on public health outcomes. Much remains to be discovered and explained about the link between low level chronic CO exposure and long-term effects on an individual’s health - for example, its impact on diseases of the cardiovascular and neurological system and whether CO is a casual factor of disease or involved in disease processes not previously associated with exposure to CO. Action is required throughout the healthcare profession, as well as by the Government, its Agencies, and Academia, to help protect the public from accidental CO poisoning

Physiological characteristics of female soccer players and health and performance considerations: a narrative review

Female soccer has seen a substantial rise in participation, as well as increased financial support from governing bodies over the last decade. Thus, there is an onus on researchers and medical departments to develop a better understanding of the physical characteristics and demands, and the health and performance needs of female soccer players. In this review, we discuss the current research, as well as the knowledge gaps, of six major topics: physical demands, talent identification, body composition, injury risk and prevention, health and nutrition. Data on female talent identification are scarce, and future studies need to elucidate the influence of relative age and maturation selection across age groups. Regarding the physical demands, more research is needed on the pattern of high-intensity sprinting during matches and the contribution of soccer-specific movements. Injuries are not uncommon in female soccer players, but targeting intrinsically modifiable factors with injury prevention programmes can reduce injury rates. The anthropometric and physical characteristics of female players are heterogeneous and setting specific targets should be discouraged in youth and sub-elite players. Menstrual cycle phase may influence performance and injury risk; however, there are few studies in soccer players. Nutrition plays a critical role in health and performance and ensuring adequate energy intake remains a priority. Despite recent progress, there is considerably less research in female than male soccer players. Many gaps in our understanding of how best to develop and manage the health and performance of female soccer players remain

Generation of a Tn5 transposon library in Haemophilus parasuis and analysis by transposon-directed insertion-site sequencing (TraDIS).

Haemophilus parasuis is an important respiratory tract pathogen of swine and the etiological agent of Glässer's disease. The molecular pathogenesis of H. parasuis is not well studied, mainly due to the lack of efficient tools for genetic manipulation of this bacterium. In this study we describe a Tn5-based random mutagenesis method for use in H. parasuis. A novel chloramphenicol-resistant Tn5 transposome was electroporated into the virulent H. parasuis serovar 5 strain 29755. High transposition efficiency of Tn5, up to 10(4) transformants/μg of transposon DNA, was obtained by modification of the Tn5 DNA in the H. parasuis strain HS071 and establishment of optimal electrotransformation conditions, and a library of approximately 10,500 mutants was constructed. Analysis of the library using transposon-directed insertion-site sequencing (TraDIS) revealed that the insertion of Tn5 was evenly distributed throughout the genome. 10,001 individual mutants were identified, with 1561 genes being disrupted (69.4% of the genome). This newly-developed, efficient mutagenesis approach will be a powerful tool for genetic manipulation of H. parasuis in order to study its physiology and pathogenesis