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Posterior Reversible Encephalopathy Syndrome (PRES) is a rare complication of Takayasu's Arteritis. A 54-year-old, right-handed woman presented with Lilliputian visual hallucinations, postprandial abdominal pain, blurred vision and headaches. She then had a tonic-clonic seizure. Neuroimaging revealed characteristic white matter oedema of the occipital lobes, in keeping with PRES. Renal infarcts and abnormalities of the abdominal aorta, subclavian, mesenteric, and internal carotid arteries were demonstrated on further imaging. The combination of hypertension, absent peripheral pulses, postprandial claudication, and imaging abnormalities of the aorta as well as its branches, lead to the diagnosis of PRES secondary to Takayasu's Arteritis. Treatment with oral steroids resulted in complete resolution of the patient's symptoms and abnormalities found on CT and MRI brain imaging. Takayasu's Arteritis is a rare vasculitis, more common in women and PRES is an unusual complication. Symptoms of PRES may include headache, seizures, hallucinations, confusion, and altered consciousness. Risk factors for PRES include; pregnancy, immunosuppression, renal disease, hypertension and rheumatological disorders. Vasogenic oedema in affected lobes, most often occipital, is characteristic of PRES on neuroimaging. Prompt treatment of PRES can avoid catastrophic consequences such as death and can achieve complete resolution of symptoms and imaging abnormalities.

We report 2 cases of obturator internus muscle strains. The injuries occurred in young male athletes involved in kicking sports. Case 1 details an acute obturator internus muscle strain with associated adductor longus strain. Case 2 details an overuse injury of the bilateral obturator internus muscles. In each case, magnetic resonance imaging played a crucial role in accurate diagnosis.

Objective: Audit is a recognised tool for evaluating the performance and improving the quality of health services. In Ireland and the UK, clear resources are available outlining audit elements. This study was undertaken to evaluate paediatric audits published from 2007 to 2020 to determine the adherence level to the definition of audit and to assess the quality of audit standards., Design: PUBMED, MEDLINE and CINAHL databases were searched to identify relevant articles published in the English language. Each was reviewed to assess whether the following criteria were met: (1) a paediatric healthcare topic was described, (2) practice was reviewed, (3) the standard was specified, (4) an intervention was made and data collection was repeated to assess improvement. The quality of the standard for each true audit was graded utilising the Oxford Centre for Evidence-Based Medicine Levels of Evidence., Results: Of 1230 published paediatric healthcare articles reviewed, 144 (11.4%) fulfilled the full criteria of an audit. Sixty-three (43.8%) true audits used the highest quality of evidence (level 1a and 1b), predominantly international or national guidelines. Fifty-six (38.9%) audits used the lowest quality of evidence (level 5), predominantly expert opinion., Conclusions: There is a mismatch between the common usage of the term audit, and the definition, despite its incorporation into training curricula and institutional support. Many articles published as audits do not adhere to the definition of audit. There are variable levels of evidence supporting the standards utilised in published true audits., (© 2022. The Author(s), under exclusive licence to Royal Academy of Medicine in Ireland.)

Recruitment and retention of doctors is a priority for the Irish healthcare service, with many leaving to work in regions with more favourable conditions. Aligning flexible training options with other jurisdictions may be an effective means of improving working conditions. We sought to assess possible improvements to the existing system and to review barriers to flexible training. We carried out a survey using 'Survey Monkey' and disseminated it to 1557 basic specialist (BST) and higher specialist trainee (HST) doctors of the Institute of Medicine, 3500 members of the Irish Medical Organisation (IMO), and across social media. There were 854 respondents; 303 (35.5%) BST, 352 (41.2%) HST, 125 (14.6%) non-training doctors, unknown, n = 74. The response rate was approximately 15-23%. Non-consultant doctors identified a preference for access to flexible training (n = 849, 99.4%), with 82.2 of doctors reporting that they would consider applying (n = 702). Most (92.4%) considered the current provision of 16 whole-time equivalent positions as inadequate (n = 789). Of doctors who would not apply for flexible training, themes identified included a perceived negative impact on their career, not meeting eligibility criteria, prolonged training, and salary implications. Suggestions for improving the system included expanding the number of places available, removing eligibility criteria, job sharing options, and the provision of regional training schemes. Access to flexible training should be a priority for the healthcare service, which may enhance recruitment and retention. A majority of our sample of non-consultant doctors identified a preference for access to flexible training options., (© 2022. The Author(s), under exclusive licence to Royal Academy of Medicine in Ireland.)

The segmented RNA genome of influenza A viruses (IAVs) enables viral evolution through genetic reassortment after multiple IAVs coinfect the same cell, leading to viruses harboring combinations of eight genomic segments from distinct parental viruses. Existing data indicate that reassortant genotypes are not equiprobable; however, the low throughput of available virology techniques does not allow quantitative analysis. Here, we have developed a high-throughput single-cell droplet microfluidic system allowing encapsulation of IAV-infected cells, each cell being infected by a single progeny virion resulting from a coinfection process. Customized barcoded primers for targeted viral RNA sequencing enabled the analysis of 18,422 viral genotypes resulting from coinfection with two circulating human H1N1pdm09 and H3N2 IAVs. Results were highly reproducible, confirmed that genetic reassortment is far from random, and allowed accurate quantification of reassortants including rare events. In total, 159 out of the 254 possible reassortant genotypes were observed but with widely varied prevalence (from 0.038 to 8.45%). In cells where eight segments were detected, all 112 possible pairwise combinations of segments were observed. The inclusion of data from single cells where less than eight segments were detected allowed analysis of pairwise cosegregation between segments with very high confidence. Direct coupling analysis accurately predicted the fraction of pairwise segments and full genotypes. Overall, our results indicate that a large proportion of reassortant genotypes can emerge upon coinfection and be detected over a wide range of frequencies, highlighting the power of our tool for systematic and exhaustive monitoring of the reassortment potential of IAVs.

Tuberculosis (TB), caused by Mycobacterium tuberculosis ( M. tb ), is a leading cause of death due to infectious disease. TB is not traditionally associated with biofilms, but M. tb biofilms are linked with drug and immune tolerance and there is increasing recognition of their contribution to the recalcitrance of TB infections. Here, we used M. tb experimental evolution to investigate this complex phenotype and identify candidate loci controlling biofilm formation. We identified novel candidate loci, adding to our understanding of the genetic architecture underlying M. tb biofilm development. Under selective pressure to grow as a biofilm, regulatory mutations rapidly swept to fixation and were associated with changes in multiple traits, including extracellular matrix production, cell size, and growth rate. Genetic and phenotypic paths to enhanced biofilm growth varied according to the genetic background of the parent strain, suggesting that epistatic interactions are important in M. tb adaptation to changing environments., Competing Interests: TS, JK, MM, SF, LB, TM, MO The authors declare no conflict of interes, MY, CP No competing interests declared, (© 2022, Smith, Youngblom et al.)

Introduction: Postgraduate medical training incorporates education, both formal and informal, combined with clinical service. This study explored the early training experience of pediatricians in Ireland and its potential impact on patient safety., Aim: We sought to identify factors that contribute to the patient safety experience of new entrant pediatric trainees., Methods: Trainees, or senior house officers (SHOs), in their first year of postgraduate training, participated in an interview conducted using a critical interview technique (CIT). They described an adverse event where the medical care delivered to the patient was not ideal. Thematic analysis identified themes that influenced the described event., Results: Thirteen trainees participated in the interviews. This study identified influences on the relationship between the SHO and patient safety, including the SHO themselves, teamwork and communication. Colleagues within the workplace, including consultants, registrars, and nurses, also affect this relationship. The registrar is described as a central figure holding an active role in clinical care in 11 of the 13 stories told. In the participants' experience, the registrar was the senior decision-maker, teacher, team builder, and communication intermediary within the teams' hierarchical structure. The registrars' previous clinical experience, communication style, along with their ability to supervise and provide feedback shaped the SHO experience., Conclusions: Through a process designed to focus on exploring patient safety, it emerged that the registrar plays a crucial role in the working experience of their junior colleagues. The influence of the registrar needs to be recognized within clinical teams and by postgraduate training bodies., (© 2021. Royal Academy of Medicine in Ireland.)

Introduction: The acquisition of non-contaminated urine samples in pre-continent infants remains a challenge. The Quick Wee method uses bladder stimulation to induce voiding. A previous randomized trial showed a higher rate of voiding within 5 minutes using this method. We evaluated this method in an Irish hospital providing secondary care., Methods: A non-blinded, randomized, controlled trial was carried out. Eligible infants were between 1 and 12 months of age, who required urine sampling as part of clinical care. Participants were randomly allocated to receive the intervention (Quick Wee Method-supra-pubic stimulation with cold saline) or the control (usual care-clean catch with no bladder stimulation) for 5 min. Primary outcome was voiding of urine within 5 min., Results: A total of 140 infants were included in this study (73 in intervention group; 67 in control group). Baseline characteristics were similar. 25% in the intervention group passed urine in the 5-min trial period compared with 18% in the control group [P = 0.4, absolute difference 7% (95% confidence interval: - 7% to + 20%)]., Conclusion: The Quick Wee method is a simple and inexpensive intervention that did not show a statistically significant increase in urine samples obtained in pre-continent infants., (© 2021. The Author(s).)

There is considerable inter-individual and inter-population variability in response to viruses. The potential of monocytes to elicit type-I interferon responses has attracted attention to their role in viral infections. Here, we use single-cell RNA-sequencing to characterize the role of cellular heterogeneity in human variation of monocyte responses to influenza A virus (IAV) exposure. We show widespread inter-individual variability in the percentage of IAV-infected monocytes. Notably, individuals with high cellular susceptibility to IAV are characterized by a lower activation at basal state of an IRF/STAT-induced transcriptional network, which includes antiviral genes such as IFITM3 , MX1 and OAS3 . Upon IAV challenge, we find that cells escaping viral infection display increased mRNA expression of type-I interferon stimulated genes and decreased expression of ribosomal genes, relative to both infected cells and those never exposed to IAV. We also uncover a stronger resistance of CD16 + monocytes to IAV infection, together with CD16 + -specific mRNA expression of IL6 and TNF in response to IAV. Finally, using flow cytometry and bulk RNA-sequencing across 200 individuals of African and European ancestry, we observe a higher number of CD16 + monocytes and lower susceptibility to IAV infection among monocytes from individuals of African-descent. Based on these data, we hypothesize that higher basal monocyte activation, driven by environmental factors and/or weak-effect genetic variants, underlies the lower cellular susceptibility to IAV infection of individuals of African ancestry relative to those of European ancestry. Further studies are now required to investigate how such cellular differences in IAV susceptibility translate into population differences in clinical outcomes and susceptibility to severe influenza., Competing Interests: JP was employed by DIACCURATE. The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 O’Neill, Quach, Pothlichet, Aquino, Bisiaux, Zidane, Deschamps, Libri, Hasan, Zhang, Zhang, Matuozzo, Cobat, Abel, Casanova, Naffakh, Rotival and Quintana-Murci.)

Aim This study evaluated the use of Lumbar Puncture (LP) in a general paediatric unit over a 3-year period. Methods Index patients, who had a successful LP, were identified from the microbiology database and failed LP procedures were identified from a chart review of the serum PCR database. Data abstracted included 1) patient age, 2) LP indication, 3) LP procedure outcome; classified as atraumatic, traumatic or failed, 4) grade of doctor undertaking the procedure and 5) the final diagnosis. Results We identified 104 paediatric patients, of whom 29(27.9%) were neonates. LP was indicated for the evaluation of acute undifferentiated illnesses, with 33 (31.7%) patients having fever without source beyond the neonatal period and 16 (15.4%) being neonates with fever. A CSF sample was obtained in 96 (92.4%) patients, with 71 (73.9%) being atraumatic. Successful LP was undertaken by Consultants in 4 (4.1%), Registrars in 83 (86.5%) and SHOs in 9 (9.4%) patients. 14 (14.6%) patients had positive CSF cultures with an additional 23 having positive cultures or serology (9 blood cultures, 11 urine cultures and 3 positive serum PCR). Conclusion Skill in LP performance is still required, to evaluate acute undifferentiated illness, in general paediatric units and ancillary methods to aid SHOs with LP skill development is desirable., Competing Interests: The authors declare that they have no conflicts of interest.

Competing Interests: None declared

Selective pressures imposed by pathogens have varied among human populations throughout their evolution, leading to marked inter-population differences at some genes mediating susceptibility to infectious and immune-related diseases. Here, we investigated the evolutionary history of a common polymorphism resulting in a Y529 versus C529 change in the cadherin related family member 3 (CDHR3) receptor which underlies variable susceptibility to rhinovirus-C infection and is associated with severe childhood asthma. The protective variant is the derived allele and is found at high frequency worldwide (69-95%). We detected genome-wide significant signatures of natural selection consistent with a rapid increase of the haplotypes carrying the allele, suggesting that non-neutral processes have acted on this locus across all human populations. However, the allele has not fixed in any population despite multiple lines of evidence suggesting that the mutation predates human migrations out of Africa. Using an approximate Bayesian computation method, we estimate the age of the mutation while explicitly accounting for past demography and positive or frequency-dependent balancing selection. Our analyses indicate a single emergence of the mutation in anatomically modern humans ~150 000 years ago and indicate that balancing selection has maintained the beneficial allele at high equilibrium frequencies worldwide. Apart from the well-known cases of the MHC and ABO genes, this study provides the first evidence that negative frequency-dependent selection plausibly acted on a human disease susceptibility locus, a form of balancing selection compatible with typical transmission dynamics of communicable respiratory viruses that might exploit CDHR3., (© The Author(s) 2019. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oup.com.)

Tsetse-transmitted African trypanosomes must develop into mammalian-infectious metacyclic cells in the fly's salivary glands (SGs) before transmission to a new host. The molecular mechanisms that underlie this developmental process, known as metacyclogenesis, are poorly understood. Blocking the few metacyclic parasites deposited in saliva from further development in the mammal could prevent disease. To obtain an in-depth perspective of metacyclogenesis, we performed single-cell RNA sequencing (scRNA-seq) from a pool of 2,045 parasites collected from infected tsetse SGs. Our data revealed three major cell clusters that represent the epimastigote, and pre- and mature metacyclic trypanosome developmental stages. Individual cell level data also confirm that the metacyclic pool is diverse, and that each parasite expresses only one of the unique metacyclic variant surface glycoprotein (mVSG) coat protein transcripts identified. Further clustering of cells revealed a dynamic transcriptomic and metabolic landscape reflective of a developmental program leading to infectious metacyclic forms preadapted to survive in the mammalian host environment. We describe the expression profile of proteins that regulate gene expression and that potentially play a role in metacyclogenesis. We also report on a family of nonvariant surface proteins (Fam10) and demonstrate surface localization of one member (named SGM1.7) on mature metacyclic parasites. Vaccination of mice with recombinant SGM1.7 reduced parasitemia early in the infection. Future studies are warranted to investigate Fam10 family proteins as potential trypanosome transmission blocking vaccine antigens. Our experimental approach is translationally relevant for developing strategies to prevent other insect saliva-transmitted parasites from infecting and causing disease in mammalian hosts., Competing Interests: Competing interest statement: The authors declare that a Provisional Patent Application describing the contents of this manuscript has been filed, Yale ref: OCR7835 || Saul ref: 047162-7268P1(01042)., (Copyright © 2020 the Author(s). Published by PNAS.)

Mycobacterium tuberculosis (M.tb) is a globally distributed, obligate pathogen of humans that can be divided into seven clearly defined lineages. An emerging consensus places the origin and global dispersal of M.tb within the past 6,000 years: identifying how the ancestral clone of M.tb spread and differentiated within this timeframe is important for identifying the ecological drivers of the current pandemic. We used Bayesian phylogeographic inference to reconstruct the migratory history of M.tb in Africa and Eurasia and to investigate lineage specific patterns of spread from a geographically diverse sample of 552 M.tb genomes. Applying evolutionary rates inferred with ancient M.tb genome calibration, we estimated the timing of major events in the migratory history of the pathogen. Inferred timings contextualize M.tb dispersal within historical phenomena that altered patterns of connectivity throughout Africa and Eurasia: trans-Indian Ocean trade in spices and other goods, the Silk Road and its predecessors, the expansion of the Roman Empire, and the European Age of Exploration. We found that Eastern Africa and Southeast Asia have been critical in the dispersal of M.tb. Our results further reveal that M.tb populations have grown through range expansion, as well as in situ, and delineate the independent evolutionary trajectories of bacterial subpopulations underlying the current pandemic., (© 2019 The Authors Molecular Ecology Published by John Wiley & Sons Ltd.)

Genetic differences are hypothesized to underlie ethnic disparities in incidence rates of the endemic systemic mycoses, including blastomycosis. Individuals of Hmong ancestry display elevated risk for this serious fungal infection. Here, we interrogated the genomes of Wisconsin (WI) Hmong blastomycosis patients using homozygosity mapping to uncover regions of the genome that are likely shared among the greater Hmong population and filtered for variants with high potential to affect disease susceptibility. This approach uncovered 113 candidate susceptibility variants, and among the most promising are those in genes involved in the interleukin-17 (IL-17) response. In particular, we identified 25 linked variants near the gene encoding IL-6 ( IL6 ). We validated differences in cytokine production between Hmong and European volunteers and formally demonstrated a critical role for IL-6 in the development of adaptive immunity to Blastomyces dermatitidis Our findings suggest that the dysregulation of IL-17 responses underlies a recently reported and poorly understood ethnic health disparity. IMPORTANCE Blastomycosis is a potentially life-threatening infection caused by the fungus Blastomyces dermatitidis As with related fungal diseases, blastomycosis is noted to affect some populations more than others. These patterns of illness are often not related to predisposing conditions or exposure risks; thus, genetic differences are thought to underlie these health disparities. People of Hmong ancestry in Wisconsin are at elevated risk of blastomycosis compared to the general population. We studied the genetic codes of Hmong blastomycosis patients and identified candidate sites in their genomes that may explain their susceptibility to this infection. We further studied one particular region of the genome that is involved with the immune processes that fight B. dermatitidis Our work revealed population differences in the response to fungi. A better understanding of the genetic underpinnings of susceptibility to infectious diseases has broader implications for community health, especially in the paradigm of personalized medicine., (Copyright © 2019 Merkhofer et al.)

[This corrects the article DOI: 10.1371/journal.ppat.1007470.].

Tsetse flies (Glossina spp.) vector pathogenic trypanosomes (Trypanosoma spp.) in sub-Saharan Africa. These parasites cause human and animal African trypanosomiases, which are debilitating diseases that inflict an enormous socio-economic burden on inhabitants of endemic regions. Current disease control strategies rely primarily on treating infected animals and reducing tsetse population densities. However, relevant programs are costly, labor intensive and difficult to sustain. As such, novel strategies aimed at reducing tsetse vector competence require development. Herein we investigated whether Kosakonia cowanii Zambiae (Kco_Z), which confers Anopheles gambiae with resistance to Plasmodium, is able to colonize tsetse and induce a trypanosome refractory phenotype in the fly. Kco_Z established stable infections in tsetse's gut and exhibited no adverse effect on the fly's survival. Flies with established Kco_Z infections in their gut were significantly more refractory to infection with two distinct trypanosome species (T. congolense, 6% infection; T. brucei, 32% infection) than were age-matched flies that did not house the exogenous bacterium (T. congolense, 36% infected; T. brucei, 70% infected). Additionally, 52% of Kco_Z colonized tsetse survived infection with entomopathogenic Serratia marcescens, compared with only 9% of their wild-type counterparts. These parasite and pathogen refractory phenotypes result from the fact that Kco_Z acidifies tsetse's midgut environment, which inhibits trypanosome and Serratia growth and thus infection establishment. Finally, we determined that Kco_Z infection does not impact the fecundity of male or female tsetse, nor the ability of male flies to compete with their wild-type counterparts for mates. We propose that Kco_Z could be used as one component of an integrated strategy aimed at reducing the ability of tsetse to transmit pathogenic trypanosomes., Competing Interests: The authors have declared that no competing interests exist.

On the basis of population genomic and phylogeographic analyses of 1669 Mycobacterium tuberculosis lineage 4 (L4) genomes, we find that dispersal of L4 has been completely dominated by historical migrations out of Europe. We demonstrate an intimate temporal relationship between European colonial expansion into Africa and the Americas and the spread of L4 tuberculosis (TB). Markedly, in the age of antibiotics, mutations conferring antimicrobial resistance overwhelmingly emerged locally (at the level of nations), with minimal cross-border transmission of resistance. The latter finding was found to reflect the relatively recent emergence of these mutations, as a similar degree of local restriction was observed for susceptible variants emerging on comparable time scales. The restricted international transmission of drug-resistant TB suggests that containment efforts at the level of individual countries could be successful.

Arthropod vectors have multiple physical and immunological barriers that impede the development and transmission of parasites to new vertebrate hosts. These barriers include the peritrophic matrix (PM), a chitinous barrier that separates the blood bolus from the midgut epithelia and modulates vector-pathogens interactions. In tsetse flies, a sleeve-like PM is continuously produced by the cardia organ located at the fore- and midgut junction. African trypanosomes, Trypanosoma brucei, must bypass the PM twice; first to colonize the midgut and secondly to reach the salivary glands (SG), to complete their transmission cycle in tsetse. However, not all flies with midgut infections develop mammalian transmissible SG infections-the reasons for which are unclear. Here, we used transcriptomics, microscopy and functional genomics analyses to understand the factors that regulate parasite migration from midgut to SG. In flies with midgut infections only, parasites fail to cross the PM as they are eliminated from the cardia by reactive oxygen intermediates (ROIs)-albeit at the expense of collateral cytotoxic damage to the cardia. In flies with midgut and SG infections, expression of genes encoding components of the PM is reduced in the cardia, and structural integrity of the PM barrier is compromised. Under these circumstances trypanosomes traverse through the newly secreted and compromised PM. The process of PM attrition that enables the parasites to re-enter into the midgut lumen is apparently mediated by components of the parasites residing in the cardia. Thus, a fine-tuned dialogue between tsetse and trypanosomes at the cardia determines the outcome of PM integrity and trypanosome transmission success.

Human-pathogenic bacteria are found in a variety of niches, including free-living, zoonotic, and microbiome environments. Identifying bacterial adaptations that enable invasive disease is an important means of gaining insight into the molecular basis of pathogenesis and understanding pathogen emergence. Staphylococcus saprophyticus , a leading cause of urinary tract infections, can be found in the environment, food, animals, and the human microbiome. We identified a selective sweep in the gene encoding the Aas adhesin, a key virulence factor that binds host fibronectin. We hypothesize that the mutation under selection ( aas _2206A>C) facilitates colonization of the urinary tract, an environment where bacteria are subject to strong shearing forces. The mutation appears to have enabled emergence and expansion of a human-pathogenic lineage of S. saprophyticus . These results demonstrate the power of evolutionary genomic approaches in discovering the genetic basis of virulence and emphasize the pleiotropy and adaptability of bacteria occupying diverse niches. IMPORTANCE Staphylococcus saprophyticus is an important cause of urinary tract infections (UTI) in women; such UTI are common, can be severe, and are associated with significant impacts to public health. In addition to being a cause of human UTI, S. saprophyticus can be found in the environment, in food, and associated with animals. After discovering that UTI strains of S. saprophyticus are for the most part closely related to each other, we sought to determine whether these strains are specially adapted to cause disease in humans. We found evidence suggesting that a mutation in the gene aas is advantageous in the context of human infection. We hypothesize that the mutation allows S. saprophyticus to survive better in the human urinary tract. These results show how bacteria found in the environment can evolve to cause disease.

In the training process, there is a tension between the work life and home life of trainees. This study explored both the personal impact and the opportunity costs of training from the Specialist Paediatric Registrar (SPR) perspective. The survey explored 1) career progression, 2) perceived functional effectiveness at work, 3) psychological impact of hospital based training, and 4) the personal and social cost of training. Fifty-three (71%) SPRs responded of whom 47 (89%)were married or in long term relationships. Seventy-five percent of trainees had a definite career plan with 86% intending to undertake fellowship training. Seventy percent believed they were efficient time managers but 53% had difficulty in making time for academic pursuits and fifty percent experienced negative feelings, which lingered after work and interfered with their relationships at home. Seventy-four percent stated training was undertaken at significant personal cost with only 21% achieving a very satisfactory work/life balance. To address these difficulties trainee wellbeing should be addressed at the Basic Specialist Training (BST) level and the career path clearly explained outlining the challenges that are likely to be encountered.

This study was undertaken to evaluate uncertainty from the Basic Specialist Trainee perspective. The survey of trainees explored 1) factors in decision making, 2) the personal impact of uncertainty, 3) the responses to both clinical errors and challenges to their decision making and 4) the potential strategies to address uncertainty. Forty-one (93%) of trainees surveyed responded. Important factors in decision making were clinical knowledge and senior colleague's opinion. Sixty percent experienced significant anxiety post call as a consequence of their uncertainty. When errors are made by colleagues, the trainee's response is acceptance (52.5%), and sympathy (32%).Trainees are strongly influenced by the opinions of senior colleagues often changing their opinions having made confident decisions. Solutions to address uncertainty include enhanced knowledge translation, and to a lesser extent, enhanced personal awareness and resilience awareness. To enhance the training experience for BST and lessen the uncertainty experienced these strategies need to be enacted within the training milieu.

Paediatric patients represent a large percentage of Emergency Department (ED) visits and there is often a perception that the acuity of these presentations is low. The decision-making process that results in parents attending the ED is poorly understood. We designed a cross-sectional cohort study to explore the reasons for attendance, the treatment initiated at home and to assess parental perception of the economic cost of attendance. Data was collected on 200 patients using a survey administered to parents in ED with a follow-up phone call 4-6 weeks later. Our results suggest that attendances are often prompted by parental anxiety rather than clinical deterioration and that prior ED usage is common among those presenting for assessment. Many parents had attempted community therapy with 128/200 patients (64%) having been referred by a healthcare professional and medical therapy at home having been employed by 114/200 (57%) parents before attendance. Parental knowledge of the safety of over-the-counter medications was variable the economic cost of an ED visit was poorly understood by participants. The results of our study suggest that parental desire for control over worrisome symptoms drives much of their management strategy prior to hospital attendance. Strategies in the ED may need to focus more on managing parental expectations than on managing the illness itself and management strategies employed should focus not only on medical therapy of the child's illness but on educating and empowering the parent.

Objective: Clinical examination of the knee is subject to measurement error. The aim of this analysis was to determine interobserver and intraobserver reliability of commonly used clinical tests in patients with knee osteoarthritis (OA)., Methods: We studied subjects with symptomatic knee OA who were participants in an open-label clinical trial of intraarticular steroid therapy. Following standardization of the clinical test procedures, 2 clinicians assessed 25 subjects independently at the same visit, and the same clinician assessed 88 subjects over an interval period of 2-10 weeks; in both cases prior to the steroid intervention. Clinical examination included assessment of bony enlargement, crepitus, quadriceps wasting, knee effusion, joint-line and anserine tenderness, and knee range of movement (ROM). Intraclass correlation coefficients (ICC), estimated kappa (κ), weighted kappa (κω), and Bland-Altman plots were used to determine interobserver and intraobserver levels of agreement., Results: Using Landis and Koch criteria, interobserver κ scores were moderate for patellofemoral joint (κ = 0.53) and anserine tenderness (κ = 0.48); good for bony enlargement (κ = 0.66), quadriceps wasting (κ = 0.78), crepitus (κ = 0.78), medial tibiofemoral joint tenderness (κ = 0.76), and effusion assessed by ballottement (κ = 0.73) and bulge sign (κω = 0.78); and excellent for lateral tibiofemoral joint tenderness (κ = 1.00), flexion (ICC = 0.97), and extension (ICC = 0.87) ROM. Intraobserver κ scores were moderate for lateral tibiofemoral joint tenderness (κ = 0.60); good for crepitus (κ = 0.78), effusion assessed by ballottement test (κ = 0.77), patellofemoral joint (κ = 0.66), medial tibiofemoral joint (κ = 0.64), and anserine tenderness (κ = 0.73); and excellent for effusion assessed by bulge sign (κω = 0.83), bony enlargement (κ = 0.98), quadriceps wasting (κ = 0.83), flexion (ICC = 0.99), and extension (ICC = 0.96) ROM., Conclusion: Among individuals with symptomatic knee OA, the reliability of clinical examination of the knee was at least good for the majority of clinical signs of knee OA., Competing Interests: Declarations of interest None

Unlabelled: This cross-sectional survey explored paediatric physician perspectives regarding diagnostic errors. All paediatric consultants and specialist registrars in Ireland were invited to participate in this anonymous online survey. The response rate for the study was 54 % (n = 127). Respondents had a median of 9-year clinical experience (interquartile range (IQR) 4-20 years). A diagnostic error was reported at least monthly by 19 (15.0 %) respondents. Consultants reported significantly less diagnostic errors compared to trainees (p value = 0.01). Cognitive error was the top-ranked contributing factor to diagnostic error, with incomplete history and examination considered to be the principal cognitive error. Seeking a second opinion and close follow-up of patients to ensure that the diagnosis is correct were the highest-ranked, clinician-based solutions to diagnostic error. Inadequate staffing levels and excessive workload were the most highly ranked system-related and situational factors. Increased access to and availability of consultants and experts was the most highly ranked system-based solution to diagnostic error., Conclusion: We found a low level of self-perceived diagnostic error in an experienced group of paediatricians, at variance with the literature and warranting further clarification. The results identify perceptions on the major cognitive, system-related and situational factors contributing to diagnostic error and also key preventative strategies., What Is Known: • Diagnostic errors are an important source of preventable patient harm and have an estimated incidence of 10-15 %. • They are multifactorial in origin and include cognitive, system-related and situational factors. What is New: • We identified a low rate of self-perceived diagnostic error in contrast to the existing literature. • Incomplete history and examination, inadequate staffing levels and excessive workload are cited as the principal contributing factors to diagnostic error in this study.

Methylmalonic acid (MMA) is a by-product of propionic acid metabolism through the vitamin B12 (cobalamin)-dependent enzyme methylmalonyl CoA mutase. Elevated MMA concentrations are a hallmark of several inborn errors of metabolism and indicators of cobalamin deficiency in older persons. In a genome-wide analysis of 2,210 healthy young Irish adults (median age 22 years) we identified a strong association of plasma MMA with SNPs in 3-hydroxyisobutyryl-CoA hydrolase (HIBCH, p = 8.42 × 10(-89)) and acyl-CoA synthetase family member 3 (ACSF3, p = 3.48 × 10(-19)). These loci accounted for 12% of the variance in MMA concentration. The most strongly associated SNP (HIBCH rs291466; c:2T>C) causes a missense change of the initiator methionine codon (minor-allele frequency = 0.43) to threonine. Surprisingly, the resulting variant, p.Met1?, is associated with increased expression of HIBCH mRNA and encoded protein. These homozygotes had, on average, 46% higher MMA concentrations than methionine-encoding homozygotes in young adults with generally low MMA concentrations (0.17 [0.14-0.21] μmol/L; median [25(th)-75(th) quartile]). The association between MMA levels and HIBCH rs291466 was highly significant in a replication cohort of 1,481 older individuals (median age 79 years) with elevated plasma MMA concentrations (0.34 [0.24-0.51] μmol/L; p = 4.0 × 10(-26)). In a longitudinal study of 185 pregnant women and their newborns, the association of this SNP remained significant across the gestational trimesters and in newborns. HIBCH is unique to valine catabolism. Studies evaluating flux through the valine catabolic pathway in humans should account for these variants. Furthermore, this SNP could help resolve equivocal clinical tests where plasma MMA values have been used to diagnose cobalamin deficiency., (Copyright © 2016 The American Society of Human Genetics. Published by Elsevier Inc. All rights reserved.)

Prevention of bacterial colonization and consequent biofilm formation remains a major challenge in implantable medical devices. Implant-associated infections are not only a major cause of implant failures but also their conventional treatment with antibiotics brings further complications due to the escalation in multidrug resistance to a variety of bacterial species. Owing to their unique properties, antimicrobial peptides (AMPs) have gained significant attention as effective agents to combat colonization of microorganisms. These peptides have been shown to exhibit a wide spectrum of activities with specificity to a target cell while having a low tendency for developing bacterial resistance. Engineering biomaterial surfaces that feature AMP properties, therefore, offer a promising approach to prevent implant infections. Here, we engineered a chimeric peptide with bifunctionality that both forms a robust solid-surface coating while presenting antimicrobial property. The individual domains of the chimeric peptides were evaluated for their solid-binding kinetics to titanium substrate as well as for their antimicrobial properties in solution. The antimicrobial efficacy of the chimeric peptide on the implant material was evaluated in vitro against infection by a variety of bacteria, including Streptococcus mutans, Staphylococcus. epidermidis, and Escherichia coli, which are commonly found in oral and orthopedic implant related surgeries. Our results demonstrate significant improvement in reducing bacterial colonization onto titanium surfaces below the detectable limit. Engineered chimeric peptides with freely displayed antimicrobial domains could be a potential solution for developing infection-free surfaces by engineering implant interfaces with highly reduced bacterial colonization property.

Tuberculosis (TB) is a global public health emergency. Increasingly drug resistant strains of Mycobacterium tuberculosis (M.tb) continue to emerge and spread, highlighting adaptability of this pathogen. Most studies of M.tb evolution have relied on 'between-host' samples, in which each person with TB is represented by a single M.tb isolate. However, individuals with TB commonly harbor populations of M.tb numbering in the billions. Here, we use analyses of M.tb genomic data from within and between hosts to gain insight into influences shaping genetic diversity of this pathogen. We find that the amount of M.tb genetic diversity harbored by individuals with TB can vary dramatically, likely as a function of disease severity. Surprisingly, we did not find an appreciable impact of TB treatment on M.tb diversity. In examining genomic data from M.tb samples within and between hosts with TB, we find that genes involved in the regulation, synthesis, and transportation of immunomodulatory cell envelope lipids appear repeatedly in the extremes of various statistical measures of diversity. Many of these genes have been identified as possible targets of selection in other studies employing different methods and data sets. Taken together, these observations suggest that M.tb cell envelope lipids are targets of selection within hosts. Many of these lipids are specific to pathogenic mycobacteria and, in some cases, human-pathogenic mycobacteria. We speculate that rapid adaptation of cell envelope lipids is facilitated by functional redundancy, flexibility in their metabolism, and their roles mediating interactions with the host.

Background: Serious bacterial illness (SBI) presents a diagnostic challenge in febrile infants. History, clinical signs, and laboratory information combined with experiential knowledge affects decisions to admit and treat., Aim: To assess the utility of serum procalcitonin and the Acute Infantile Observation Score (AIOS) performed at emergency department presentation in predicting (a) confirmed serious bacterial illness and (b) illness severity., Methods: Sensitivity, specificity, and likelihood ratios were calculated for C-reactive protein, white cell count, serum procalcitonin, and AIOS., Results: Forty-six infants were recruited. Seven had a diagnosis of SBI, 28 were moderately ill with length of stay >24 hours, and 12 were severely ill with length of stay >96 hours. The positive likelihood ratios for confirmed SBI were C-reactive protein = 5.3, procalcitonin = 0.43, white cell count = 1.9, and AIOS = 1.5., Conclusion: Procalcitonin and the AIOS do not modify the diagnostic uncertainty of the ED physician assessing the febrile infant with respect to admission or antibiotic treatment above standard laboratory investigations.

Consultant paediatricians in Ireland were surveyed to evaluate their perceptions of the hospital audit environment and assess their involvement in the audit process. Eighty nine (77%) replied of whom 66 (74%) had an audit department and 23 (26%) did not. Sixteen (18%) felt their hospital was well resourced for audit and 25 (28%) felt the culture was very positive but only 1 (1%) had protected time. For 61 (69%) consultants audit was very important with 38 (43%) being very actively involved in the process. The most frequent trigger for audit was non consultant hospital doctor (NCHD) career development, cited by 77 (87%). The new Professional Competence Scheme and the National Quality and Risk Management Standards will require the deficiencies identified in this survey be addressed.

Children who appear healthy, even if they have one or more recognized cardiovascular risk factors, do not generally have outcomes of cardiovascular or other vascular disease during childhood. Historically, pediatric medicine has not aggressively screened for or treated cardiovascular risk factors in otherwise healthy children. However, studies such as the P-Day Study (Pathobiological Determinants of Atherosclerosis in Youth), and the Bogalusa Heart Study, indicate that healthy children at remarkably young ages can have evidence of significant atherosclerosis. With the increasing prevalence of pediatric obesity, can we expect more health problems related to the consequences of pediatric dyslipidemia, hypertriglyceridemia, and atherosclerosis in the future? For many years, medications have been available and used in adult populations to treat dyslipidemia. In recent years, reports of short-term safety of some of these medications in children have been published. However, none of these studies have detailed long-term follow-up, and therefore none have described potential late side-effects of early cholesterol-lowering therapy, or potential benefits in terms of reduction of or delay in cardiovascular or other vascular end-points. In 2007, the American Heart Association published a scientific statement on the use of cholesterol-lowering therapy in pediatric patients. In this review paper, we discuss some of the current literature on cholesterol-lowering therapy in children, including the statins that are currently available for use in children, and some of the cautions with using these and other cholesterol-lowering medications. A central tenet of this review is that medications are not a substitute for dietary and lifestyle interventions, and that even in children on cholesterol-lowering medications, physicians should take every opportunity to encourage children and their parents to make healthy diet and lifestyle choices.

Heterozygous familial hypercholesterolemia (heFH) affects 1 in 500 individuals. Evidence supports the low-density lipoprotein (LDL)-lowering effect of statins for adults with heFH. However, there are concerns regarding the treatment children with heFH. By performing a systematic review and metaanalysis of the published literature, this study aimed to evaluate the efficacy and safety of statins used for children with heFH. A systematic review was performed by searching multiple medical databases and citations to identify reports of randomized controlled trials of statins used to treat children with heFH. The trials were retrieved, reviewed, and subjected to metaanalysis. The search yielded 2,174 titles. Of the 63 studies retrieved and reviewed, 56 were excluded, 7 were included in the systematic review, and 4 were included in the metaanalysis. Significant heterogeneity was detected. The metaanalysis showed significant LDL lowering, high-density lipoprotein (HDL) cholesterol elevation, and increases in height and weight with statins. The metaanalysis could not be performed for many side effects of statins, but individual trials showed no significant side effects. Quality assessment showed methodologic concerns, with potential for bias. For example, six trials analyzed statin effects without intention to treat despite such a stated intention. Metaanalysis shows significant LDL lowering with statin treatment. Further studies, including epidemiologic and multicenter studies, are required.

Human dihydrofolate reductase (DHFR) is a critical target in cancer chemotherapy. Previous studies showed that an 82-nt RNA fragment within the DHFR mRNA protein-coding region functions as a DHFR cis-acting response element. In this study, we further investigated the key elements contained within this sequence that are required for the DHFR mRNA-DHFR protein interaction. Using enzymatic foot-printing assays and RNA-binding experiments, we isolated a 27-nt sequence (DHFR27, corresponding to nts 407-433), which bound with high affinity and specificity to human DHFR to form a ribonucleoprotein complex. In vivo transient transfection experiments using a luciferase reporter system revealed that DHFR27 RNA could repress the luciferase expression in a DHFR-dependent manner when placed upstream of luciferase mRNA. This work provides new insights into the essential molecular elements that mediate RNA-protein interactions.

Elevation of low-density lipoprotein (LDL) cholesterol is a recognized risk factor for premature atherosclerosis and such elevations have their antecedents in the pediatric population. This study evaluated the frequency of elevated LDL cholesterol in children and adolescents (8-18 years) in whom one parent had a history of premature ischemic heart disease (<55 years). Patients had a fasting lipid profile performed and results were classified as normal, borderline/high, or high in accordance with the American Academy of Pediatrics (AAP) guidelines. Fifty-two parents had a total of 97 children and adolescents who were eligible for screening. Ninety-three initially consented and fasting lipid profiles were obtained in 82 (84.5%) children. Prevalences of high and borderline/high LDL cholesterol were (7/82) 8.5% and (17/82) 20.7%, respectively. Screening for hypercholesterolemia in these at-risk children and adolescents is prudent and dietary advice must be offered.

Aim: To evaluate the quality of paediatric audits from 1966 to 1999., Methods: A Medline search was performed using the MeSH terms audit, child, paediatric (and pediatric). Predefined core elements of audit were used as inclusion criteria for entry of an article into this study. These criteria were as follows: (1) an article deals with a healthcare topic; (2) a standard is predefined; (3) actual practice is evaluated; (4) actual practice is compared with the standard. The fifth criterion of audit, dissemination of information and reaudit, was not an inclusion criterion, as it was not used in the early years covered by this study. Empirical grading of standards was used., Results: The search yielded 442 articles, of which 303 (100%) were related to paediatric healthcare and were reviewed. Standards were defined in 115 (38%) articles. Audit against the standard was performed in 92 (30.4%) articles, of which 42 (45.6%) were published before, and 50 (54.3%) after, 1990. 18 (5.9%) articles were re-audited: 6 (14.3%) were published before, and 12 (24%) after, 1990. Of the 188 paediatric studies rejected, 119 (63.3%) described practice observations., Conclusion: Many articles in paediatrics are published as "audits", but they do not contain the core elements of audit. Although audit is a potentially valuable tool in clinical medicine, the publication of poor-quality audits may lead to the decline of the audit concept. Suggestions on ways to improve the quality of published audits are made.

To achieve asthma control adequate knowledge of the disease is required. Knowledge levels of thirty five specialist registrars and one hundred and sixty parents of children attending an asthmatic clinic, prior to entry into a formalised asthma educational program, were assessed utilising a twenty stem questionnaire with eighty five statements. Three out of thirty five specialist registrars had adequate knowledge levels whilst the other thirty two had potentially adequate knowledge levels. Sixty two out of one hundred and sixty parents had potentially adequate knowledge levels and the other ninety eight had inadequate knowledge levels. Parental knowledge can be improved by formal educational programs. Deficiencies in doctors' knowledge should be addressed through a similar program rather than rely on experiential learning.

All Irish paediatric higher specialist trainees' opinions about the paediatric higher specialist training (HST) scheme and related manpower issues were surveyed. Information was obtained on 1) trainees' level of satisfaction with HST, 2) their ultimate career ambitions including location of final posts, 3) attitudes to both flexible training and consultancies and 4) demographics to assess the significance of gender variations. Fifty-two eligible trainees were identified using the Royal College of Physicians of Ireland database. The survey was administered as an anonymous postal survey. The response rate was 88%. Results indicated a high level of satisfaction with HST (78%) overall although problems were noted with the half-day release programme as only 63% were facilitated. Only 30% wish to practice as subspecialists, 76% of trainees wish to work in an urban hospital and 43.5% desire a flexible consultancy suggesting an incompatibility of trainees' desires with current Irish medical manpower policy. To address these difficulties we suggest establishing more rigorous audit of training posts to ensure deficiencies are corrected and the establishment of flexible training to address gender imbalance and to promote the concept of consultant job sharing.