bluuebunny/crossref_metadata_2025_split huggingface dataset:
ThefruRBAOperon Is Necessary for Group A Streptococcal Growth in Fructose and for Resistance to Neutrophil Killing during Growth in Whole Human Blood
Authors: Valdes Kayla M. , Sundar Ganesh S. , Vega Luis A. , Belew Ashton T. , Islam Emrul , Binet Rachel , El-Sayed Najib M. , Le Breton Yoann , McIver Kevin S. 
Published: January 2016

ABSTRACTBacterial pathogens rely on the availability of nutrients for survival in the host environment. The phosphoenolpyruvate-phosphotransferase system (PTS) is a global regulatory network connecting sugar uptake with signal transduction. Since the fructose PTS has been shown to impact virulence in several streptococci, including the human pathogenStreptococcus pyogenes(the group AStreptococcus[GAS]), we characterized its role in carbon metabolism and pathogenesis in the M1T1 strain 5448. Growth in fructose as a sole carbon source resulted in 103 genes affected transcriptionally, where thefrulocus (fruRBA) was the most induced. Reverse transcriptase PCR showed thatfruRBAformed an operon which was repressed by FruR in the absence of fructose, in addition to being under carbon catabolic repression. Growth assays and carbon utilization profiles revealed that although the entirefruoperon was required for growth in fructose, FruA was the main transporter for fructose and also was involved in the utilization of three additional PTS sugars: cellobiose, mannitol, andN-acetyl-d-galactosamine. The inactivation ofsloR, afruAhomolog that also was upregulated in the presence of fructose, failed to reveal a role as a secondary fructose transporter. Whereas the ability of both ΔfruRand ΔfruBmutants to survive in the presence of whole human blood or neutrophils was impaired, the phenotype was not reproduced in murine whole blood, and those mutants were not attenuated in a mouse intraperitoneal infection. Since the ΔfruAmutant exhibited no phenotype in the human or mouse assays, we propose that FruR and FruB are important for GAS survival in a human-specific environment.
Tissue-Resident T Cells as the Central Paradigm of Chlamydia Immunity
Authors: Johnson Raymond M. , Brunham Robert C. 
Published: January 2016

ABSTRACTFor almost 2 decades, results fromChlamydiapathogenesis investigations have been conceptualized using a cytokine polarization narrative. Recent viral immunity studies identifying protective tissue-resident memory T cells (Trm) suggest an alternative paradigm based on localized immune networks. AsChlamydiavaccines enter the preclinical pipeline and, in the case of an attenuated trachoma vaccine, are given to human subjects, it may be useful to ask whether cytokine polarization is the appropriate framework for understanding and evaluating vaccine efficacy. In this review, we revisitC. trachomatispathogenesis data from mice and humans using a Trm narrative and note a comfortable concordance with theChlamydiapathogenesis literature.
Genes Required for the Fitness of Salmonella enterica Serovar Typhimurium during Infection of Immunodeficientgp91−/−phoxMice
Authors: Grant Andrew J. , Oshota Olusegun , Chaudhuri Roy R. , Mayho Matthew , Peters Sarah E. , Clare Simon , Maskell Duncan J. , Mastroeni Pietro 
Published: January 2016

ABSTRACTSalmonella entericacauses systemic diseases (typhoid and paratyphoid fever), nontyphoidal septicemia (NTS), and gastroenteritis in humans and other animals worldwide. An important but underrecognized emerging infectious disease problem in sub-Saharan Africa is NTS in children and immunocompromised adults. A current goal is to identifySalmonellamutants that are not pathogenic in the absence of key components of the immune system such as might be found in immunocompromised hosts. Such attenuated strains have the potential to be used as live vaccines. We have used transposon-directed insertion site sequencing (TraDIS) to screen mutants ofSalmonella entericaserovar Typhimurium for their ability to infect and grow in the tissues of wild-type and immunodeficient mice. This was to identify bacterial genes that might be deleted for the development of live attenuated vaccines that would be safer to use in situations and/or geographical areas where immunodeficiencies are prevalent. The relative fitness of each of 9,356 transposon mutants, representing mutations in 3,139 different genes, was determined ingp91−/−phoxmice. Mutations in certain genes led to reduced fitness in both wild-type and mutant mice. To validate these results, these genes were mutated by allelic replacement, and resultant mutants were retested for fitness in the mice. A defined deletion mutant ofcysEwas attenuated in C57BL/6 wild-type mice and immunodeficientgp91−/−phoxmice and was effective as a live vaccine in wild-type mice.
Deficiency of the Complement Component 3 but Not Factor B Aggravates Staphylococcus aureus Septic Arthritis in Mice
Authors: Na Manli , Jarneborn Anders , Ali Abukar , Welin Amanda , Magnusson Malin , Stokowska Anna , Pekna Marcela , Jin Tao 
Published: January 2016

ABSTRACTThe complement system plays an essential role in the innate immune response and protection against bacterial infections. However, detailed knowledge regarding the role of complement inStaphylococcus aureusseptic arthritis is still largely missing. In this study, we elucidated the roles of selected complement proteins inS. aureusseptic arthritis. Mice lacking the complement component 3 (C3−/−), complement factor B (fB−/−), and receptor for C3-derived anaphylatoxin C3a (C3aR−/−) and wild-type (WT) control mice were intravenously or intra-articularly inoculated withS. aureusstrain Newman. The clinical course of septic arthritis, as well as histopathological and radiological changes in joints, was assessed. After intravenous inoculation, arthritis severity and frequency were significantly higher inC3−/−mice than in WT controls, whereasfB−/−mice displayed intermediate arthritis severity and frequency. This was in accordance with both histopathological and radiological findings. C3, but not fB, deficiency was associated with greater weight loss, more frequent kidney abscesses, and higher bacterial burden in kidneys.S. aureusopsonized withC3−/−sera displayed decreased uptake by mouse peritoneal macrophages compared with bacteria opsonized with WT orfB−/−sera. C3aR deficiency had no effect on the course of hematogenousS. aureusseptic arthritis. We conclude that C3 deficiency increases susceptibility to hematogenousS. aureusseptic arthritis and impairs host bacterial clearance, conceivably due to diminished opsonization and phagocytosis ofS. aureus.
Sibling Rivalry in Myxococcus xanthus Is Mediated by Kin Recognition and a Polyploid Prophage
Authors: Dey Arup , Vassallo Christopher N. , Conklin Austin C. , Pathak Darshankumar T. , Troselj Vera , Wall Daniel 
Published: January 2016

ABSTRACT

Myxobacteria form complex social communities that elicit multicellular behaviors. One such behavior is kin recognition, in which cells identify siblings via their polymorphic TraA cell surface receptor, to transiently fuse outer membranes and exchange their contents. In addition, outer membrane exchange (OME) regulates behaviors, such as inhibition of wild-type
Myxococcus xanthus
(DK1622) from swarming. Here we monitored the fate of motile cells and surprisingly found they were killed by nonmotile siblings. The kill phenotype required OME (i.e., was TraA dependent). The genetic basis of killing was traced to ancestral strains used to construct DK1622. Specifically, the kill phenotype mapped to a large “polyploid prophage,” Mx alpha. Sensitive strains contained a 200-kb deletion that removed two of three Mx alpha units. To explain these results, we suggest that Mx alpha expresses a toxin-antitoxin cassette that uses the OME machinery of
M. xanthus
to transfer a toxin that makes the population “addicted” to Mx alpha. Thus, siblings that lost Mx alpha units (no immunity) are killed by cells that harbor the element. To test this, an Mx alpha-harboring laboratory strain was engineered (by
traA
allele swap) to recognize a closely related species,
Myxococcus fulvus
. As a result,
M. fulvus
, which lacks Mx alpha, was killed. These TraA-mediated antagonisms provide an explanation for how kin recognition specificity might have evolved in myxobacteria. That is, recognition specificity is determined by polymorphisms in
traA
, which we hypothesize were selected for because OME with non-kin leads to lethal outcomes.

IMPORTANCE
The transition from single cell to multicellular life is considered a major evolutionary event. Myxobacteria have successfully made this transition. For example, in response to starvation, individual cells aggregate into multicellular fruiting bodies wherein cells differentiate into spores. To build fruits, cells need to recognize their siblings, and in part, this is mediated by the TraA cell surface receptor. Surprisingly, we report that TraA recognition can also involve sibling killing. We show that killing originates from a prophage-like element that has apparently hijacked the TraA system to deliver a toxin to kin. We hypothesize that this killing system has imposed selective pressures on kin recognition, which in turn has resulted in TraA polymorphisms and hence many different recognition groups.
Functional Determinants of Metal Ion Transport and Selectivity in Paralogous Cation Diffusion Facilitator Transporters CzcD and MntE in Streptococcus pneumoniae
Authors: Martin Julia E. , Giedroc David P. 
Published: January 2016

ABSTRACT

Cation diffusion facilitators (CDFs) are a large family of divalent metal transporters that collectively possess broad metal specificity and contribute to intracellular metal homeostasis and virulence in bacterial pathogens.
Streptococcus pneumoniae
expresses two homologous CDF efflux transporters, MntE and CzcD. Cells lacking
mntE
or
czcD
are sensitive to manganese (Mn) or zinc (Zn) toxicity, respectively, and specifically accumulate Mn or Zn, respectively, thus suggesting that MntE selectively transports Mn, while CzcD transports Zn. Here, we probe the origin of this metal specificity using a phenotypic growth analysis of pneumococcal variants. Structural homology to
Escherichia coli
YiiP predicts that both MntE and CzcD are dimeric and each protomer harbors four pairs of conserved metal-binding sites, termed the A site, the B site, and the C1/C2 binuclear site. We find that single amino acid mutations within both the transmembrane domain A site and the B site in both CDFs result in a cellular metal sensitivity similar to that of the corresponding null mutants. However, multiple mutations in the predicted cytoplasmic C1/C2 cluster of MntE have no impact on cellular Mn resistance, in contrast to the analogous substitutions in CzcD, which do have on impact on cellular Zn resistance. Deletion of the MntE-specific C-terminal tail, present only in Mn-specific bacterial CDFs, resulted in only a modest growth phenotype. Further analysis of MntE-CzcD functional chimeric transporters showed that Asn and Asp in the
N
D-
D
D A-site motif of MntE and the most N-terminal His in the
H
D-HD site A of CzcD (the specified amino acids are underlined) play key roles in transporter metal selectivity.

IMPORTANCE
Cation diffusion facilitator (CDF) proteins are divalent metal ion transporters that are conserved in organisms ranging from bacteria to humans and that play important roles in cellular physiology, from metal homeostasis and resistance to type I diabetes in vertebrates. The respiratory pathogen
Streptococcus pneumoniae
expresses two metal CDF transporters, CzcD and MntE. How CDFs achieve metal selectivity is unclear. We show here that CzcD and MntE are true paralogs, as CzcD transports zinc, while MntE selectively transports manganese. Through the use of an extensive collection of pneumococcal variants, we show that a primary determinant for metal selectivity is the A site within the transmembrane domain. This extends our understanding of how CDFs discriminate among transition metals.
Identification of a Functional Plasmodesmal Localization Signal in a Plant Viral Cell-To-Cell-Movement Protein
Authors: Yuan Cheng , Lazarowitz Sondra G. , Citovsky Vitaly 
Published: January 2016

ABSTRACT

Our fundamental knowledge of the protein-sorting pathways required for plant cell-to-cell trafficking and communication via the intercellular connections termed plasmodesmata has been severely limited by the paucity of plasmodesmal targeting sequences that have been identified to date. To address this limitation, we have identified the plasmodesmal localization signal (PLS) in the
Tobacco mosaic virus
(TMV) cell-to-cell-movement protein (MP), which has emerged as the paradigm for dissecting the molecular details of cell-to-cell transport through plasmodesmata. We report here the identification of a bona fide functional TMV MP PLS, which encompasses amino acid residues between positions 1 and 50, with residues Val-4 and Phe-14 potentially representing critical sites for PLS function that most likely affect protein conformation or protein interactions. We then demonstrated that this PLS is both necessary and sufficient for protein targeting to plasmodesmata. Importantly, as TMV MP traffics to plasmodesmata by a mechanism that is distinct from those of the three plant cell proteins in which PLSs have been reported, our findings provide important new insights to expand our understanding of protein-sorting pathways to plasmodesmata.

IMPORTANCE
The science of virology began with the discovery of
Tobacco mosaic virus
(TMV). Since then, TMV has served as an experimental and conceptual model for studies of viruses and dissection of virus-host interactions. Indeed, the TMV cell-to-cell-movement protein (MP) has emerged as the paradigm for dissecting the molecular details of cell-to-cell transport through the plant intercellular connections termed plasmodesmata. However, one of the most fundamental and key functional features of TMV MP, its putative plasmodesmal localization signal (PLS), has not been identified. Here, we fill this gap in our knowledge and identify the TMV MP PLS.
Summative service and stakeholder evaluation of an NHS-funded community Pharmacy Emergency Repeat Medication Supply Service (PERMSS)
Authors: Nazar Hamde , Nazar Zachariah , Simpson Jill , Yeung Andre , Whittlesea Cate 
Published: January 2016

ObjectivesService and stakeholder evaluation of an NHS-funded service providing out-ofhours (OOH) emergency repeat medications to patients self-presenting at community pharmacies.SettingCommunity pharmacies across the North East of England accredited to provide this service.ParticipantsPatients self-presenting to community pharmacies during OOH periods with emergency repeat medication supply requests.InterventionCommunity pharmacists assessed each request for clinical appropriateness and when suitable provide an emergency repeat medication supply, with additional pharmaceutical advice and services if required.Primary outcomesNumber of emergency repeat medication supplies, time of request, reason for access, medication(s), pharmaceutical advice and services provided. Secondary outcomes were community pharmacist and patient satisfaction.ResultsA total of 2485 patients were managed across 227 community pharmacies (15 December 2014 to 7 April 2015). Most patients presented on Saturdays, with increased activity over national holidays. Older age was associated with increased service use. Of the 3226 medications provided, 439 were classified as high risk. Patients found this service easy to access and were willing to access the community pharmacy in the future for medication-related issues. In the absence of this service, 50% of patients would have missed their medication(s) until they saw their doctor and a further 46% would have accessed an alternative service. The cost of National Health Service (NHS) service(s) for patients who would have accessed an alternative OOH service was estimated as 37 times that of the community pharmacy service provided. Community pharmacists were happy to provide this service despite increased consultation times and workload.ConclusionsCommunity pharmacists were able to manage patients’ OOH requests for emergency repeat medication and patients were happy with the service provided. Since the service cost was favourable when compared with alternative OOH services, it would be a viable option to reduce the workload on the wider NHS.
Attitudes and beliefs about deceased organ donation in the Arabic-speaking community in Australia: a focus group study
Authors: Ralph Angelique F , Alyami Ali , Allen Richard D M , Howard Kirsten , Craig Jonathan C , Chadban Steve J , Irving Michelle , Tong Allison 
Published: January 2016

ObjectivesTo describe the beliefs and attitudes to organ donation in the Arabic-speaking community.DesignArabic-speaking participants were purposively recruited to participate in 6 focus groups. Transcripts were analysed thematically.Participants53 participants, aged 19–77 years, and originating from 8 countries, participated in 1 of 6 focus groups. Participants identified as Christian (73%), Islam (26%), Buddhist (2%) or did not identify with any religion (2%).Results6 themes (with subthemes) were identified; religious conviction; invisibility of organ donation; medical suspicion; owning the decision; and reciprocal benefit.ConclusionsAlthough organ donation is considered a generous life-saving ‘gift’, representative members of the Arabic-speaking community in Australia were unfamiliar with, unnerved by and sceptical about the donation process. Making positive decisions about organ donation would likely require resolving tensions between respecting family, community and religious values versus their individual autonomy. Providing targeted education about the process and benefits of organ donation within the Arabic community may clarify ambiguities surrounding cultural and religious-based views on organ donation, reduce taboos and suspicion towards donation, and in turn, lead to increased organ donation rates.
The sensitivity and specificity of clinical measures of sport concussion: three tests are better than one
Authors: Resch Jacob E , Brown Cathleen N , Schmidt Julianne , Macciocchi Stephen N , Blueitt Damond , Cullum C Munro , Ferrara Michael S 
Published: January 2016

ContextA battery of clinical measures of neurocognition, balance and symptoms has been recommended for the management of sport concussion (SC) but is based on variable evidence.ObjectiveTo examine the sensitivity and specificity of a battery of tests to assess SC in college athletes.DesignCross-sectional.SettingResearch laboratory.Patients or other participantsDivision 1 athletes diagnosed with a SC (n=40) who were 20.2±1.60 years of age and 180.5±11.12 cm tall and healthy athletes (n=40) who were 19.0±0.93 years of age and 179.1±11.39 cm tall were enrolled.Intervention(s)Participants were administered Immediate Postconcussion Assessment and Cognitive Test (ImPACT), the Sensory Organization Test (SOT) and the Revised Head Injury Scale (HIS-r) prior to and up to 24 h following injury between the 2004 and 2014 sport seasons. Sensitivity and specificity were calculated using predictive discriminant analyses (PDA) and clinical interpretation guidelines.Main outcome measuresOutcome measures included baseline and postinjury ImPACT, SOT and HIS-r composite scores.ResultsUsing PDA, each clinical measure's sensitivity ranged from 55.0% to 77.5% and specificity ranged from 52.5% to 100%. The test battery possessed a sensitivity and specificity of 80.0% and 100%, respectively. Using clinical interpretation guidelines, sensitivity ranged from 55% to 97.5% individually, and 100% when combined.ConclusionsOur results support a multidimensional approach to assess SC in college athletes which correctly identified 80–100% of concussed participants as injured. When each test was evaluated separately, up to 47.5% of our sample was misclassified. Caution is warranted when using singular measures to manage SC.
We have the programme, what next? Planning the implementation of an injury prevention programme
Authors: Donaldson Alex , Lloyd David G , Gabbe Belinda J , Cook Jill , Finch Caroline F 
Published: January 2016

Background and aimThe impact of any injury prevention programme is a function of the programme and its implementation. However, real world implementation of injury prevention programmes is challenging. Lower limb injuries (LLIs) are common in community Australian football (community-AF) and it is likely that many could be prevented by implementing exercise-based warm-up programmes for players. This paper describes a systematic, evidence-informed approach used to develop the implementation plan for a LLI prevention programme in community-AF in Victoria, Australia.MethodsAn ecological approach, using Step 5 of the Intervention Mapping health promotion programme planning protocol, was taken.ResultsAn implementation advisory group was established to ensure the implementation plan and associated strategies were relevant to the local context. Coaches were identified as the primary programme adopters and implementers within an ecological system including players, other coaches, first-aid providers, and club and league administrators. Social Cognitive Theory was used to identify likely determinants of programme reach, adoption and implementation among coaches (eg, knowledge, beliefs, skills and environment). Diffusion of Innovations theory, the Implementation Drivers framework and available research evidence were used to identify potential implementation strategies including the use of multiple communication channels, programme resources, coach education and mentoring.ConclusionsA strategic evidence-informed approach to implementing interventions will help maximise their population impact. The approach to implementation planning described in this study relied on an effective researcher-practitioner partnership and active engagement of stakeholders. The identified implementation strategies were informed by theory, evidence and an in-depth understanding of the implementation context.
Marked socioeconomic inequalities in hip fracture incidence rates during the Bone and Joint Decade (2000–2010) in Portugal: age and sex temporal trends in a population based study
Authors: Oliveira Carla Maria , Alves Sandra Maria , Pina Maria Fátima 
Published: January 2016

BackgroundSocioeconomic factors may influence changes in hip fracture (HF) incidence over time. We analysed HF temporal trends during the Bone and Joint Decade in Portugal (BJD-Portugal), 2000–2010, by regional socioeconomic status (SES), sex and age.MethodsWe selected registers of patients aged 50+ years with HF (International Classification of Diseases, V.9—Clinical Modification, ICD9-CM) caused by traumas of low/moderate energy, from the National Hospital Discharge Database. Annual time series of age-specific incidence rates were calculated by sex and regional SES (deprived, medium, affluent). Generalised additive models were fitted to identify shape/turning points in temporal trends.ResultsWe selected 96 905 HF (77.3% in women). Women were older than men at admission (81.2±8.5 vs 78.2±10.1 years-old, p<0.001). For women 65–79 years, a continuously decreasing trend (1.7%/year) only in affluent and increasing trends (3.3–3.4%/year) after 2006/2007 in medium and deprived was observed. For men, trends were stable or increased in almost all age/SES groups (only two decreasing periods). For the oldest women, all SES present similar trends: turning points around 2003 (initiating decreasing periods: 1.8–2.9%/year) and around 2007 (initiating increasing periods: 3.7–3.3%/year).ConclusionsThere were SES-sex-age inequalities in temporal trends during BJD-Portugal: marked SES inequalities among women aged 65–79 years (a persistent, decreasing trend only in the affluent) vanished among the oldest women; the same was not observed in men, for them, there were almost no declining periods; women aged ≥80 years, presented increasing trends around 2007, as in most deprived/age/sex groups. Despite some successful periods of decreasing trends, incidence rates did not improve overall in almost all age groups and both sexes.