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.

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.

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.

Activation pathways of synovial T lymphocytes. Expression and function of the UM4D4/CDw60 antigen.

Accumulating evidence implicates a central role for synovial T cells in the pathogenesis of rheumatoid arthritis, but the activation pathways that drive proliferation and effector function of these cells are not known. We have recently generated a novel monoclonal antibody against a rheumatoid synovial T cell line that recognizes an antigen termed UM4D4 (CDw60). This antigen is expressed on a minority of peripheral blood T cells, and represents the surface component of a distinct pathway of human T cell activation. The current studies were performed to examine the expression and function of UM4D4 on T cells obtained from synovial fluid and synovial membranes of patients with rheumatoid arthritis and other forms of inflammatory joint disease. The UM4D4 antigen is expressed at high surface density on about three-fourths of synovial fluid T cells and on a small subset of synovial fluid natural killer cells; in synovial tissue it is present on more than 90% of T cells in lymphoid aggregates, and on approximately 50% of T cells in stromal infiltrates In addition, UM4D4 is expressed in synovial tissue on a previously undescribed population of HLA-DR/DP-negative non-T cells with a dendritic morphology. Anti-UM4D4 was co-mitogenic for both RA and non-RA synovial fluid mononuclear cells, and induced IL-2 receptor expression. The UM4D4/CDw60 antigen may represent a functional activation pathway for synovial compartment T cells, which could play an important role in the pathogenesis of inflammatory arthritis.

MRI in osteoarthritis of the hip: gradations of severity.

In a prospective study, 10 patients with well-documented osteoarthritis (O.A.) of the hips were imaged using spin-echo pulse sequences (TR = 0.5 to 1.5 s and TE = 28 to 60 ms). After analyzing the changes observed, an MR grading system for assessing severity of O.A. in the hips was developed. Using this grading system and an established grading system for osteoarthritis using roentgenograms (both systems use grades 0-4), two radiologists independently graded the MR studies and plain films separately, twice. The roentgenogram grading system was more accurate in predicting symptoms in the more severe cases, whereas the MR grading system was slightly more useful in the less severe cases. Our results show that MR can demonstrate a spectrum of changes of O.A. in the hips. Its ability to directly image articular cartilage makes it a powerful research and clinical tool.

Quality control in continuing education.

A quality continuing education programme may be seen as one which is of a high standard, relevant and viable. Efforts made to ensure the quality of continuing education should be directed towards these three ends. Where possible learners should be involved in programme planning, implementation and evaluation and should be encouraged to put their learning into practice. This paper will explore the issues and practicalities involved in attempts to ensure the quality of educational activities provided for qualified professionals by groups or bodies who are concerned with the professional development and quality of service of both individuals and professional groups.

An in-service training programme for intern physiotherapists.

In-service education is one component or method of continuing education. This paper discusses the development of an in-service training programme for intern physiotherapists which is being conducted at the Prince Henry and Prince of Wales Hospitals in Sydney. The basic philosophy behind the programme is the proposition that the intern year for physiotherapists should be both a year of service and a total learning experience aimed at providing a transition between student training and independent professional responsibilities and capabilities. Accordingly the programme was planned to provide training for the interns which would assist them in developing clinical expertise in the tasks of a junior physiotherapist. The programme is proving to be a useful component of the overall learning experiences available to the 19 intern physiotherapists who are allocated to these hospitals each year.

The role of intermittent positive pressure breathing in respiratory physiotherapy.

This article reviews the value and limitations of I.P.P.B. as opposed to other treatment methods available to the respiratory physiotherapist. In the treatment of patients with airways obstruction I.P.P.B. is presented as having a limited role. Alternative measures such as the use of simple nebulisation, relaxation therapy and manual methods to remove secretions are described as being preferable to I.P.P.B. in most cases. I.P.P.B. has some value in the early post-operative care of some patients undergoing surgery which results in limitation to spontaneous deep breathing postoperatively. However other methods, notably incentive spirometry should also be considered for these patients. In the treatment and prevention of respiratory complications associated with neuromuscular and skeletal disorders I.P.P.B. can play a significant role in counteracting the possible effects of hypoventilation. I.P.P.B. should be considered as one method among many which is available to the physiotherapist to assist in the treatment of respiratory complications associated with medical and surgical conditions.

Self-directed learning as part of the mainstream of physiotherapy education.

A frequently stated aim of physiotherapy educational programs is to produce graduates capable of, and wishing to pursue, lifelong learning. To help achieve this goal, curricula commonly include the promotion of independent or self-directed learning. This paper supports the need for developing self-directed learning, orientation and skills and argues that this can be best achieved by bringing these skills into the mainstream of tertiary education programs such as physiotherapy, rather than isolating them to specific subjects. The challenge of mainstreaming is to appreciate how the basic ideas of self-directed learning can be incorporated into all areas of the curriculum. This paper presents strategies, issues and problems associated with mainstreaming of self-directed learning.

Educational programmes to develop clinical reasoning skills.

The role of physiotherapists has changed considerably over the last few decades. Autonomous professionals have replaced clinicians who applied technical skills under the direction of medical practitioners. The physiotherapy profession needs more than ever to produce clinicians who demonstrate competence in clinical reasoning and decision making. The challenge presented to all physiotherapists involved in teaching is to contribute to the development of clinicians who can use the complex skill of clinical reasoning in association with a sound clinical knowledge base. This paper discusses issues and strategies associated with achieving this goal.

Designing a pulmonary rehabilitation programme.

Physiotherapists have an important role to play in the rehabilitation of patients with chronic obstructive pulmonary disease. Individually-tailored programmes including the following components, namely education, exercise, secretion removal, breathing 'training', home programmes, ventilatory muscle training, medications, support systems and counselling can provide subjective and objective benefits for these patients. A primary consideration is to involve patients in the planning, implementation and evaluation of the programme and to encourage them to develop self-help skills. Guidelines for designing a pulmonary rehabilitation programme are outlined,' and pertinent literature reviewed.

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.

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.

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.

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.