Prenatal exome sequencing and impact on perinatal outcome: cohort study.

OBJECTIVES: First, to determine the uptake of prenatal exome sequencing (pES) and the diagnostic yield of pathogenic (causative) variants in a UK tertiary fetal medicine unit following the introduction of the NHS England Rapid Exome Sequencing Service for fetal anomalies testing (R21 pathway). Second, to identify how the decision to proceed with pES and identification of a causative variant affect perinatal outcomes, specifically late termination of pregnancy (TOP) at or beyond 22 weeks' gestation. METHODS: This was a retrospective cohort study of anomalous fetuses referred to the Liverpool Women's Hospital Fetal Medicine Unit between 1 March 2021 and 28 February 2022. pES was performed as part of the R21 pathway. Trio exome sequencing was performed using an Illumina next-generation sequencing platform assessing coding and splice regions of a panel of 974 prenatally relevant genes and 231 expert reviewed genes. Data on demographics, phenotype, pES result and perinatal outcome were extracted and compared. Descriptive statistics and the χ-square or Fisher's exact test were performed using IBM SPSS version 28.0.1.0. RESULTS: In total, 72 cases were identified and two-thirds of eligible women (n = 48) consented to trio pES. pES was not feasible in one case owing to a low DNA yield and, therefore, was performed in 47 cases. In one-third of cases (n = 24), pES was not proposed or agreed. In 58.3% (14/24) of these cases, this was because invasive testing was declined and, in 41.7% (10/24) of cases, women opted for testing and underwent chromosomal microarray analysis only. The diagnostic yield of pES was 23.4% (11/47). There was no overall difference in the proportion of women who decided to have late TOP in the group in which pES was agreed compared with the group in which pES was not proposed or agreed (25.0% (12/48) vs 25.0% (6/24); P = 1.0). However, the decision to have late TOP was significantly more frequent when a causative variant was detected compared with when pES was uninformative (63.6% (7/11) vs 13.9% (5/36); P < 0.0009). The median turnaround time for results was longer in cases in which a causative variant was identified than in those in which pES was uninformative (22 days (interquartile range (IQR), 19-34) days vs 14 days (IQR, 10-15 days); P < 0.0001). CONCLUSIONS: This study demonstrates the potential impact of identification of a causative variant by pES on decision to have late TOP. As the R21 pathway continues to evolve, we urge clinicians and policymakers to consider introducing earlier screening for anomalies, developing robust guidance for late TOP and ensuring optimized support for couples. © 2022 The Authors. Ultrasound in Obstetrics & Gynecology published by John Wiley & Sons Ltd on behalf of International Society of Ultrasound in Obstetrics and Gynecology.

Genomes of Novel Myxococcota Reveal Severely Curtailed Machineries for Predation and Cellular Differentiation.

Cultured Myxococcota are predominantly aerobic soil inhabitants, characterized by their highly coordinated predation and cellular differentiation capacities. Little is currently known regarding yet-uncultured Myxococcota from anaerobic, nonsoil habitats. We analyzed genomes representing one novel order (o__JAFGXQ01) and one novel family (f__JAFGIB01) in the Myxococcota from an anoxic freshwater spring (Zodletone Spring) in Oklahoma, USA. Compared to their soil counterparts, anaerobic Myxococcota possess smaller genomes and a smaller number of genes encoding biosynthetic gene clusters (BGCs), peptidases, one- and two-component signal transduction systems, and transcriptional regulators. Detailed analysis of 13 distinct pathways/processes crucial to predation and cellular differentiation revealed severely curtailed machineries, with the notable absence of homologs for key transcription factors (e.g., FruA and MrpC), outer membrane exchange receptor (TraA), and the majority of sporulation-specific and A-motility-specific genes. Further, machine learning approaches based on a set of 634 genes informative of social lifestyle predicted a nonsocial behavior for Zodletone Myxococcota. Metabolically, Zodletone Myxococcota genomes lacked aerobic respiratory capacities but carried genes suggestive of fermentation, dissimilatory nitrite reduction, and dissimilatory sulfate-reduction (in f_JAFGIB01) for energy acquisition. We propose that predation and cellular differentiation represent a niche adaptation strategy that evolved circa 500 million years ago (Mya) in response to the rise of soil as a distinct habitat on Earth. IMPORTANCE The phylum Myxococcota is a phylogenetically coherent bacterial lineage that exhibits unique social traits. Cultured Myxococcota are predominantly aerobic soil-dwelling microorganisms that are capable of predation and fruiting body formation. However, multiple yet-uncultured lineages within the Myxococcota have been encountered in a wide range of nonsoil, predominantly anaerobic habitats, and the metabolic capabilities, physiological preferences, and capacity of social behavior of such lineages remain unclear. Here, we analyzed genomes recovered from a metagenomic analysis of an anoxic freshwater spring in Oklahoma, USA, that represent novel, yet-uncultured, orders and families in the Myxococcota. The genomes appear to lack the characteristic hallmarks for social behavior encountered in Myxococcota genomes and displayed a significantly smaller genome size and a smaller number of genes encoding biosynthetic gene clusters, peptidases, signal transduction systems, and transcriptional regulators. Such perceived lack of social capacity was confirmed through detailed comparative genomic analysis of 13 pathways associated with Myxococcota social behavior, as well as the implementation of machine learning approaches to predict social behavior based on genome composition. Metabolically, these novel Myxococcota are predicted to be strict anaerobes, utilizing fermentation, nitrate reduction, and dissimilarity sulfate reduction for energy acquisition. Our results highlight the broad patterns of metabolic diversity within the yet-uncultured Myxococcota and suggest that the evolution of predation and fruiting body formation in the Myxococcota has occurred in response to soil formation as a distinct habitat on Earth.

Improving the WHO Surgical Safety Checklist sign-out.

BACKGROUND: The WHO Surgical Safety Checklist has been shown to reduce perioperative morbidity and mortality worldwide. There is evidence to suggest that sign-out is the most poorly performed phase of the checklist as it coincides with a period of high workload for team members. This study aimed to see whether modification of this process might result in greater compliance. METHODS: A controlled longitudinal (before and after) study was performed to evaluate the effect of a modified checklist sign-out on compliance in a single surgical department. Checklist quality was evaluated by measurement of checklist completion, active participation, and team member presence. Workload assessment was performed to identify the optimal moment for the sign-out process. The sign-out process was modified through an iterative multidisciplinary approach, informed by results from the workload assessment. Feedback was obtained through staff surveys. RESULTS: A total of 185 operations were used, with an intervention group in vascular surgery and a control group in orthopaedics. The optimal timing for sign-out was identified as after final wound closure. The modified sign-out process improved active participation of team members (21 of 34 versus 31 of 34; P = 0.010). In the control group, complete compliance improved (48 of 76 versus 30 of 41; P = 0.041). However, active participation decreased (53 of 76 versus 19 of 41; P = 0.022). No differences were noted between groups in team member presence. Eighteen of 21 staff questioned viewed the modifications positively. CONCLUSION: The optimal sign-out timing was identified as immediately after final wound closure prior to undraping the patient.

The BRCA2 polymorphic stop codon: stuff or nonsense?

BACKGROUND: Despite classification of the BRCA2c.9976A>T, p.(Lys3326Ter) variant as a polymorphism, it has been associated with increased risks of pancreatic, lung, oesophageal and breast cancer. METHODS: We have noticed multiple co-occurrences of the BRCA2 c.9976A>T variant with the pathogenic BRCA2c.6275_6276delTT frameshift mutation p.(Leu2092ProfsTer7) and using a cohort study have assessed if this might account for these tumour risk associations. RESULTS: We identified 52 families with BRCA2c.6275_6276delTT, all of which occur in cis with the BRCA2c.9976A>T variant allele as demonstrated by co-segregation in all family members tested. Of 3245 breast/ovarian cancer samples sequenced for BRCA2, only 43/3245 (1.3%) carried BRCA2 c.9976A>T alone, after excluding individuals with BRCA2c.6275_6276delTT (n=22) or other BRCA1 (n=3) or BRCA2 (n=2) pathogenic mutations. The resultant frequency (1.3%) after removal of co-occurring mutations is lower than the 1.7% and 1.67% frequencies from two control populations for BRCA2 c.9976A>T, but similar to the 1.39% seen in the Exome Aggregation Consortium database. We did not identify increased frequencies of oesophageal, pancreatic or lung cancer in families with just BRCA2 c.9976A>T using person-years at risk analysis. CONCLUSIONS: It is likely that the previous associations of increased cancer risks due to BRCA2c.9976A>T represent reporting bias and are contributed to because the variant is in LD with BRCA2c.6275_6276delTT.

EAACI food allergy and anaphylaxis guidelines: managing patients with food allergy in the community.

The European Academy of Allergy and Clinical Immunology (EAACI) Food Allergy and Anaphylaxis Guidelines, managing patients with food allergy (FA) in the community, intend to provide guidance to reduce the risk of accidental allergic reactions to foods in the community. This document is intended to meet the needs of early-childhood and school settings as well as providers of non-prepackaged food (e.g., restaurants, bakeries, takeaway, deli counters, and fast-food outlets) and targets the audience of individuals with FA, their families, patient organizations, the general public, policymakers, and allergists. Food allergy is the most common trigger of anaphylaxis in the community. Providing children and caregivers with comprehensive information on food allergen avoidance and prompt recognition and management of allergic reactions are of the utmost importance. Provision of adrenaline auto-injector devices and education on how and when to use these are essential components of a comprehensive management plan. Managing patients at risk of anaphylaxis raises many challenges, which are specific to the community. This includes the need to interact with third parties providing food (e.g., school teachers and restaurant staff) to avoid accidental exposure and to help individuals with FA to make safe and appropriate food choices. Education of individuals at risk and their families, their peers, school nurses and teachers as well as restaurant and other food retail staff can reduce the risk of severe/fatal reactions. Increased awareness among policymakers may improve decision-making on legislation at local and national level.

Practitioner-client relationships in integrative medicine clinics in Australia: a contemporary social phenomenon.

OBJECTIVES: The increasing use of complementary and alternative medicine (CAM) in developed countries has been attributed more to contemporary cultural trends than to inherent problems in mainstream medicine. The aim of this study was to examine the impact of post-modern values on relationships between practitioners and clients in integrative medicine (IM) clinics in Australia. DESIGN: This research used hermeneutic phenomenology to uncover experiences of practitioner-client relationships in IM and the meanings clients and practitioners attached to these relationships. Data were collected using cumulative case studies, focus groups and key informant interviews. Data analysis consisted of reading and re-reading texts derived from interview transcripts and field notes and constantly comparing texts to identify meanings and patterns. Themes extracted from the data set were continually refined, discarded and elaborated until meta-themes emerged. SETTING: Australian IM clinics where general medical practitioners and CAM practitioners were co-located. RESULTS: Post-modern values were evident in practitioner-client relationships in Australian IM clinics and were manifested in two ways. (1) Clients did not bring an expectation that they would receive prescriptive treatment regimens. They regarded consultations as opportunities for obtaining information or advice, or for monitoring their health. (2) Practitioners valued clients' knowledge and judgments and respected clients' right to choose and direct their health care. CONCLUSIONS: In the IM clinics in this research, the traditional role of the practitioner as sole possessor of expertise had changed. Practitioners had become one among a number of resources that informed clients used when designing personal health care plans or negotiating health care with practitioners.

Familial essential thrombocythemia with spontaneous megakaryocyte colony formation and acquired JAK2 mutations.

Essential thrombocythemia (ET) is a chronic myeloproliferative disorder, characterized by increased proliferation of megakaryocytes and elevated platelet count that usually occurs sporadically. We report a family with seven affected individuals in three generations, including one individual with a phenotype resembling polycythemia vera, a related disorder. Megakaryocyte (CFU-MK) colony formation occurred in the absence of added cytokines in cultures of peripheral blood from affected family members. Some reports of familial ET have identified mutations in THPO and MPL, the genes for a cytokine (thrombopoietin, TPO) that regulates platelet production and its receptor (c-MPL), respectively. In this family, the MPL gene was excluded by linkage analysis. Although TPO levels were elevated in most affected family members and evidence for linkage was found between the disease and THPO (theta=0.0, Z(max)=3.0), a THPO mutation was not identified by DNA sequencing. The JAK2 V617F mutation that has been associated with 50% of sporadic cases of ET was identified as a somatic mutation, an acquired defect, in peripheral blood of the two most severely affected family members. These patients also had elevated TPO levels. Further study of familial myeloproliferative diseases will help elucidate the initiating genetic events underlying ET.

Multiple levels of Notch signal regulation (review).

Notch is a vitally important signalling receptor controlling cell fate determination and pattern formation in numerous ways during development of both invertebrate and vertebrate species. An intriguing pathway for the Notch signal has emerged where, after ligand-dependent proteolysis, an intracellular fragment of the receptor itself translocates to the nucleus to regulate gene expression. The nuclear activity of the Notch intracellular domain is linked to complexes regulating chromatin organization through histone deacetylation and acetylation. To allow the Notch signal to be deployed in numerous contexts, many different mechanisms have evolved to regulate the level, duration and spatial distribution of Notch activity. Regulation occurs at multiple levels including patterns of ligand and receptor expression, Notch-ligand interactions, trafficking of the receptor and ligands, and covalent modifications including glycosylation, phosphorylation and ubiquitination. Several Notch regulatory proteins have conserved domains that link them to the ubiquitination pathway, and ubiquitination of the Notch intracellular domain has recently been linked to its degradation. Different proteolytically derived isoforms of Notch have also been identified that may be involved in alternative Notch-dependent signals or regulatory mechanisms, and differences between the four mammalian Notch homologues are beginning to be appreciated.

Portrait of the physiotherapy profession.

As a profession integral to health promotion, prevention, acute care and rehabilitation physiotherapy plays an essential role in the health care system. This paper explores the nature of physiotherapy, the role of physiotherapy in health care, the practice of physiotherapy internationally, the education of physiotherapists, the regulation of physiotherapy practice and the maintenance of practice standards.

Integrating clinical reasoning and evidence-based practice.

Evidence-based practice involves basing clinical decisions and practice on the best available evidence. What do nurses need to think about when seeking to pursue evidence-based practice? What challenges do practitioners face in blending clinical reasoning with evidence-based practice? In this article, the authors argue against basing clinical practice on narrow definitions of evidence, relying solely on experimental findings or, even more exclusively, on randomized controlled trials. Instead of defining best practice narrowly by the strength of the current empirical evidence used to guide clinical decisions, it should be defined broadly by what is the best information to use to make decisions for a given patient in a particular setting. Credible and accountable clinical decisions rely on a number of forms of knowledge and evidence. This evidence includes findings from across the range of research methods, including experimental, interpretive, and action research. In nursing, it also includes evidence that has arisen from the depth of practice knowledge of expert clinical nurses. Professionals, particularly advanced and expert clinicians such as critical care nurses, use clinical reasoning to guide their practice in the uncertainty of clinical practice contexts. This reasoning helps the nurse make judgments about the relevance of particular research and clinical evidence for a specific patient and setting.

Parvovirus arthropathy outbreak in southwestern United States.

OBJECTIVE: We describe an outbreak of parvovirus (PV) arthropathy that was detected in a rheumatology clinic in San Antonio, Texas, during the winter of 1994. Parvovirus B19 causes acute symmetric polyarthritis (ASPA) in adults. In the US, the majority of cases described are from the northern US. METHODS: An outbreak of PV arthropathy was monitored in a San Antonio area rheumatology clinic. RESULTS: Of the 16 affected patients, 69% were female, ages ranging from 23 to 60 years; 75% had close contact with children, 58% of whom were exposed to children with clinical PV. All patients noted an acute arthritis except for 2 patients with polyarthralgias. The most common presentation was ASPA (9/16), with 10/16 complaining of viral prodrome, and 5/16 having a nonspecific rash, but none with the typical "slapped cheek" appearance. Eleven patients had an ASPA at some time in their illness. Of these, 3 had a true migratory arthritis that developed into an ASPA and another 2 were additive. Two additional patients had persistent asymmetric polyarthritis. The most common joints involved were the metacarpophalangeals, proximal interphalangeals, wrists, and knees. Most patients' syndromes lasted < 6 weeks, but 3 patients had symptoms that lasted longer than 6 months. Eight of 10 had elevated erythrocyte sedimentation rate. Rheumatoid factor was detected in 3 patients and antinuclear antibody in 2. All patients were treated symptomatically with nonsteroidal antiinflammatory drugs and a few also received low dose corticosteroids. Because of suspicious clinical presentations, 2 patients were presumed to have gonococcal arthritis before PV titers were available. CONCLUSION: This is the first large series on adults with PV arthropathy reported in the southern US. In contrast to the usual features of ASPA, the outbreak appears unique in that almost 40% of cases presented with a true migratory arthritis.

Preparing for the workplace: fostering generic attributes in allied health education programs.

Allied health curricula need to extend beyond the learning of discipline-specific skills to encompass broader learning goals. In particular, the acquisition of generic skills is necessary to enable graduates to function more competently and confidently within their rapidly changing work, professional, and societal environments. In health sciences education particularly, the rate of change in practice and education is rapid and unprecedented. If educators focus on components of the curriculum rather than the entire learning experience, they are likely to significantly limit the students' acquisition of such generic skills. To achieve the desired generic skills outcomes, an overarching, integrated, and consistently applied curriculum strategy is advocated. This article considers a number of such strategies relevant to allied health education.

Azathioprine hepatotoxicity is uncommon in patients with Rheumatic diseases.

At present, there is no consensus regarding the utility of liver enzyme monitoring in rheumatology patients undergoing treatment with azathioprine (AZA). To further investigate this issue, we retrospectively reviewed for current or past use of AZA all patient records in our Disease Modifying Antirheumatic Drug (DMARD) clinic from May 1986 through September 1996. Information available from individual DMARD charts included AZA data (dates initiated, dose, indication for changing dose or stopping AZA), co-administered DMARDs, and aspartate aminotransferase (AST) serum values. In patients followed in the clinic through February 1994, serum alkaline phosphatase (AP) values additionally were available. Published reports of AZA hepatotoxicity in rheumatoid arthritis (RA) patients were found by a MEDLINE search, supplemented by a manual search of reference lists.We screened 429 patient records, and all patients who had received AZA at any time during the study period (n = 56) were identified for further review. The mean daily AZA dose was 96 mg (median = 100 mg/ day, range = 25-200 mg/day). All of the 56 patients had undergone routine periodic monitoring of serum AST while taking AZA (mean interval = 26 days); 30 of these patients additionally underwent monitoring of serum AP (mean interval = 24 days). Twenty-three (41%) of the patients had at least one episode of an abnormally elevated serum AST and/or AP while taking AZA. Only 1 of these 2 laboratory values was abnormal in 12 patients. The average time from AZA initiation to first appearance of an abnormal AST and/or AP value was 27 days (range = 7-62 days). During observation alone, all abnormal laboratory test results became normalized in 20 of 23 patients (87%) during the study period. There were no interventions in response to an abnormal laboratory value, and no patient experienced an adverse clinical outcome attributable to AZA hepatotoxicity during the 10-year study period.AZA use for rheumatic conditions may be associated with elevations in both the serum AST and/or AP values. Most of these elevations were transient, and none were clinically significant in our patient population. This study supports current American College of Rheumatology guidelines, which do not recommend routine periodic monitoring with liver enzyme tests in RA patients taking AZA. Furthermore, this study suggests that the utility of the baseline liver enzyme test in these patients is not well established.

Quantitative measurement of erosion growth and joint space loss in rheumatoid arthritis hand radiographs.

OBJECTIVE: To evaluate the performance of simple, inexpensive quantitative techniques for measuring erosion growth and joint space loss in serial hand radiographs of patients with rheumatoid arthritis (RA). METHODS: Erosions were measured using a plastic overlay template of sample erosion sizes. Joint spaces were measured with a hand held measuring micrometer. In the first phase of testing, a spectrum of individual erosions and joint spaces was measured to determine intra and interobserver correlations and variability. In the 2nd phase, the tools were used to measure serial changes in RA hand radiographs. Observer correlations and the ability to discriminate serial changes were determined and compared to the scoring method of Sharp. RESULTS: Measurements of individual erosion areas and joint spaces were highly reproducible. Intra and interobserver correlations were significant (p < 0.05) for serial erosion growth and joint space loss measurements, as well as changes in Sharp scores. Quantitative measurements correlated highly with the corresponding Sharp score changes. Of all measurements or scores recorded, quantitative joint space loss measurements were statistically superior in discriminating serial change in RA hand radiographs. Quantitatively measured joint space loss correlated well with both erosion growth measurements and serial change in total Sharp scores. CONCLUSION: Quantitative measurement of erosion growth and joint space loss is possible with simple inexpensive techniques. Further study is needed to confirm our data, which suggest that quantitative measurement of joint space narrowing may be the most useful discriminator of serial changes in RA hand radiographs.

An unusual cause of thigh pain after total hip arthroplasty.

The aim of total hip arthroplasty surgery is to relieve pain. There are many postulated causes of thigh pain following total hip arthroplasty, some of which are not easily corrected. Muscle hernia as a result of hip surgery is a cause of thigh pain that is disabling, relatively easy to diagnose, and may be preventable.

Photographing the patient with burns for medical documentation.

Photographs of a burn injury provide an objective description of the patient's disease course and may be important in the medicolegal documentation of child abuse or other criminal and civil actions. Accurate and reproducible photographs require careful composition and choice of equipment. Standardized clinical positioning is described with recommendations for timing of serial photographs.

How effectively do we use double staff time?

This paper describes a group of registered nurses' investigation of the use of 'double staff time' in a neonatal unit. Using participatory action research, the nurses explored why it was difficult for the unit's staff to take part in continuing education programs that were provided during double staff time. The group drew on their personal experiences as well as data collected from peers in the neonatal unit. The data illustrated that nursing activities and nursing handovers were two factors that encroached on nurses' professional development time.

Fluconazole therapy for coccidioidal meningitis. The NIAID-Mycoses Study Group.

OBJECTIVE: To determine the efficacy and safety of fluconazole treatment of coccidioidal meningitis. DESIGN: Uncontrolled clinical trial. SETTING: Four university-based treatment centers in California, Arizona, and Texas. Most therapy was conducted without hospitalization. PATIENTS: Fifty consecutive patients with active coccidioidal meningitis, of which 47 (94%) were evaluable. Twenty-five patients had received no previous treatment for their meningitis, and nine had coinfection with human immunodeficiency virus (HIV). INTERVENTION: Fluconazole was administered in an oral dose of 400 mg once per day for up to 4 years (median, 37 months) in responding patients. Concurrent therapy with another antifungal agent was prohibited. MEASUREMENTS: Predefined assessment of infection-related abnormalities was done at the time of enrollment and was repeated at least every 4 months during treatment. Elimination of 40% or more of baseline abnormalities was considered a response. RESULTS: Thirty-seven of 47 (79%; 95% CI, 61% to 90%) evaluable patients responded to treatment. Response rates were similar for patients with and without previous therapy, for patients with and without concomitant HIV infection, and for patients with and without pre-existing hydrocephalus. Most improvement occurred within 4 to 8 months after starting treatment. Patient symptoms resolved more quickly than did cerebrospinal fluid abnormalities. In 15 of 20 responding patients followed for 20 months or more, residual low-level cerebrospinal fluid abnormalities remained throughout therapy. No patient discontinued therapy because of drug-related side effects, although confusion developed in two patients that resolved when the dose of fluconazole was reduced. CONCLUSION: Fluconazole therapy is often effective in suppressing coccidioidal meningitis.

A programme for developing clinical reasoning skills in graduate physiotherapists.

This paper presents a clinical reasoning course for postgraduate physiotherapy students, conducted at the University of Sydney. The course was designed to foster the development of students' clinical reasoning skills and clinical knowledge. The rationale and nature of the course are discussed, along with evaluation findings. Results indicate that the course has succeeded in stimulating students' interest and enjoyment in clinical reasoning and promoting students' understanding of and competence in this complex ability.