Iron starvation results in up-regulation of a probable Haloferax volcanii siderophore transporter.

The response of the haloarchaeal model organism Haloferax volcanii to iron starvation was analyzed at the proteome level by data-independent acquisition mass spectrometry. Cells grown in minimal medium with normal iron levels were compared to those grown under low iron conditions, with samples being separated into membrane and cytoplasmic fractions in order to focus on import/export processes which are frequently associated with metal homeostasis. Iron starvation not only caused a severe retardation of growth but also altered the levels of many proteins. Using a comprehensive annotated spectral library and data-independent acquisition mass spectrometry (DIA-MS), we found that iron starvation resulted in significant changes to both the membrane and the soluble proteomes of Hfx. volcanii. The most affected protein is the RND family permease HVO_A0467, which is 44-fold enriched in cells grown under iron starvation. The gene HVO_A0467 can be deleted suggesting that it is not essential under standard conditions. Compared to wild type cells the deletion strain shows only slight changes in growth and cell morphologies show no differences. Molecular docking predictions indicated that HVO_A0467 may be an exporter of the siderophore schizokinen for which a potential biosynthesis cluster is encoded in the Hfx. volcanii genome. Together, these findings confirm the importance of iron for archaeal cells and suggest HVO_0467 as a siderophore exporter.

Torque about electrostatically charged spheres makes them more attractive.

The strength of interparticle interactions in a granular system controls how a collection of insulating particles flow, cohere and fragment. Forces due to electrostatic charging, particularly in free-fall or low gravity environments, can dominate the static and dynamic interactions with important implications for understanding natural and industrial processes. Here we show that shaking of homogeneous, spherical particles can result in a non-uniform surface charge distribution. The measured dipole moment and torque for each particle are found to be strongly correlated. However, our model shows that to predict the torque and force requires one to consider the full surface charge distribution. This overlooked torque is not only significant, but would amplify attractive interactions through particle reorientation.

Lessons for Flying Astronauts with Disabilities Drawn from Experience in Aviation.

INTRODUCTION: Accessible spaceflight may seem a distant concept. As part of a diverse European Space Agency funded Topical Team, we are working on the physiological feasibility of space missions being undertaken by people with physical disabilities. Here, the first activity of this team is presented in the form of key lessons learned from aviation to inform new work on space missions.DISCUSSION: The first lesson is agreeing on realistic expectations about impairments, their severity, and the possibility of flying independently. This is important in terms of astronaut recruitment and societal expectations. The second lesson relates to training and adjustments for people with disabilities. Flexibility is important while maintaining safety for everyone involved. The third lesson is about managing unconscious bias from the different stakeholders. We conclude by arguing that engagement with people from different backgrounds is essential for the success of the first space mission with people with physical disabilities.Miller-Smith MJ, Tucker N, Anderton R, Caplin N, Harridge SDR, Hodkinson P, Narici MV, Pollock RD, Possnig C, Rittweger J, Smith TG, Di Giulio I. Lessons for flying astronauts with disabilities drawn from experience in aviation. Aerosp Med Hum Perform. 2024; 95(9):716-719.

Involvement of ArlI, ArlJ, and CirA in archaeal type IV pilin-mediated motility regulation.

Many prokaryotes use swimming motility to move toward favorable conditions and escape adverse surroundings. Regulatory mechanisms governing bacterial flagella-driven motility are well-established; however, little is yet known about the regulation underlying swimming motility propelled by the archaeal cell surface structure, the archaella. Previous research showed that the deletion of the adhesion pilins (PilA1-6), subunits of the type IV pili cell surface structure, renders the model archaeon Haloferax volcanii non-motile. In this study, we used ethyl methanesulfonate mutagenesis and a motility assay to identify motile suppressors of the ∆pilA[1-6] strain. Of the eight suppressors identified, six contain missense mutations in archaella biosynthesis genes, arlI and arlJ. In trans expression of arlI and arlJ mutant constructs in the respective multi-deletion strains ∆pilA[1-6]∆arlI and ∆pilA[1-6]∆arlJ confirmed their role in suppressing the ∆pilA[1-6] motility defect. Additionally, three suppressors harbor co-occurring disruptive missense and nonsense mutations in cirA, a gene encoding a proposed regulatory protein. A deletion of cirA resulted in hypermotility, while cirA expression in trans in wild-type cells led to decreased motility. Moreover, quantitative real-time PCR analysis revealed that in wild-type cells, higher expression levels of arlI, arlJ, and the archaellin gene arlA1 were observed in motile early-log phase rod-shaped cells compared to non-motile mid-log phase disk-shaped cells. Conversely, ∆cirA cells, which form rods during both early- and mid-log phases, exhibited similar expression levels of arl genes in both growth phases. Our findings contribute to a deeper understanding of the mechanisms governing archaeal motility, highlighting the involvement of ArlI, ArlJ, and CirA in pilin-mediated motility regulation.IMPORTANCEArchaea are close relatives of eukaryotes and play crucial ecological roles. Certain behaviors, such as swimming motility, are thought to be important for archaeal environmental adaptation. Archaella, the archaeal motility appendages, are evolutionarily distinct from bacterial flagella, and the regulatory mechanisms driving archaeal motility are largely unknown. Previous research has linked the loss of type IV pili subunits to archaeal motility suppression. This study reveals three Haloferax volcanii proteins involved in pilin-mediated motility regulation, offering a deeper understanding of motility regulation in this understudied domain while also paving the way for uncovering novel mechanisms that govern archaeal motility. Understanding archaeal cellular processes will help elucidate the ecological roles of archaea as well as the evolution of these processes across domains.

Involvement of ArlI, ArlJ, and CirA in Archaeal Type-IV Pilin-Mediated Motility Regulation.

Many prokaryotes use swimming motility to move toward favorable conditions and escape adverse surroundings. Regulatory mechanisms governing bacterial flagella-driven motility are well-established, however, little is yet known about the regulation underlying swimming motility propelled by the archaeal cell surface structure, the archaella. Previous research showed that deletion of the adhesion pilins (PilA1-6), subunits of the type IV pili cell surface structure, renders the model archaeon Haloferax volcanii non-motile. In this study, we used EMS mutagenesis and a motility assay to identify motile suppressors of the ΔpilA[1-6] strain. Of the eight suppressors identified, six contain missense mutations in archaella biosynthesis genes, arlI and arlJ. Overexpression of these arlI and arlJ mutant constructs in the respective multi-deletion strains ΔpilA[1-6]ΔarlI and ΔpilA[1-6]ΔarlJ confirmed their role in suppressing the ΔpilA[1-6] motility defect. Additionally, three suppressors harbor co-occurring disruptive missense and nonsense mutations in cirA, a gene encoding a proposed regulatory protein. A deletion of cirA resulted in hypermotility, while cirA overexpression in wild-type cells led to decreased motility. Moreover, qRT-PCR analysis revealed that in wild-type cells, higher expression levels of arlI, arlJ, and the archaellin gene arlA1 were observed in motile early-log phase rod-shaped cells compared to non-motile mid-log phase disk-shaped cells. Conversely, ΔcirA cells, which form rods during both early and mid-log phases, exhibited similar expression levels of arl genes in both growth phases. Our findings contribute to a deeper understanding of the mechanisms governing archaeal motility, highlighting the involvement of ArlI, ArlJ, and CirA in pilin-mediated motility regulation.

Halobacterium hubeiense sp. nov., a haloarchaeal species isolated from a bore core drilled in Hubei Province, PR China.

Eight colonies of live microbes were isolated from an extensively surface-sterilized halite sample which had been retrieved from a depth of 2000 m from a salt mine in the Qianjiang Depression, Hubei Province, PR China. The eight colonies, obtained after 4 weeks of incubation, were named JI20-1T-JI20-8 and JI20-1T was selected as the type strain. The strains have been previously described, including a genomic analysis based on the complete genome for strain JI20-1T and draft genomes for the other strains. In that study, the name Halobacterium hubeiense was suggested, based on the location of the drilling site. Previous phylogenomic analysis showed that strain JI20-1T is most closely related to the Permian isolate Halobacterium noricense from Alpine rock salt. The orthologous average nucleotide identity (orthoANI) and digital DNA-DNA hybridization (dDDH) percentages between the eight strains are 100-99.6 % and 99.8-96.4 %, respectively. The orthoANI and dDDH values of these strains with respect to the type strains of species of the genus Halobacterium are 89.9-78.2 % and 37.3-21.6 %, respectively, supporting their placement in a novel extremely halophilic archaeal species. The phylogenomic tree based on the comparison of sequences of 632 core-orthologous proteins confirmed the novel species status for these haloarchaea. The polar lipid profile includes phosphatidylglycerol, phosphatidylglycerol phosphate methyl ester, phosphatidylglycerol sulfate, and sulfated galactosyl mannosyl galactosyl glucosyl diether, a profile compatible with that of Halobacterium noricense. Based on genomic, phenotypic, and chemotaxonomic characterization, we propose strain JI20-1T (=DSM 114402T = HAMBI 3616T) as the type strain of a novel species in the genus Halobacterium, with the name Halobacterium hubeiense sp. nov.

Proposed minimal standards for description of new taxa of the class Halobacteria.

Halophilic archaea of the class Halobacteria are the most salt-requiring prokaryotes within the domain Archaea. In 1997, minimal standards for the description of new taxa in the order Halobacteriales were proposed. From then on, the taxonomy of the class Halobacteria provides an excellent example of how changing concepts on prokaryote taxonomy and the development of new methods were implemented. The last decades have witnessed a rapid expansion of the number of described taxa within the class Halobacteria coinciding with the era of genome sequencing development. The current members of the International Committee on Systematics of Prokaryotes Subcommittee on the Taxonomy of Halobacteria propose these revisions to the recommended minimal standards and encourage the use of advanced technologies in the taxonomic description of members of the Halobacteria. Most previously required and some recommended minimal standards for the description of new taxa in the class Halobacteria were retained in the present revision, but changes have been proposed in line with the new methodologies. In addition to the 16S rRNA gene, the rpoB' gene is an important molecular marker for the identification of members of the Halobacteria. Phylogenomic analysis based on concatenated conserved, single-copy marker genes is required to infer the taxonomic status of new taxa. The overall genome relatedness indexes have proven to be determinative in the classification of the taxa within the class Halobacteria. Average nucleotide identity, digital DNA-DNA hybridization, and average amino acid identity values should be calculated for rigorous comparison among close relatives.

Developing a web-based app for non-mental health nurses to assess the mental health needs and risks of children and young people.

There is high demand for specialist mental health services for children and young people in the UK. Non-mental health nurses are well-placed to assess the mental health needs and risks of children and young people to maximise opportunities for early intervention and relieve the pressure on child and adolescent mental health services. This article provides an overview of a service development project to develop a web-based application (app) to support non-mental health nurses when assessing the mental health needs and risks of children and young people. The article describes the development, testing and evaluation process, which involved consultation with children and young people as well as interviews, focus groups and an online survey with a range of professionals working with children and young people. Overall, the findings suggest that the app is appropriate for use by non-mental health nurses in terms of quality, functionality and acceptability.

Interactome Analysis Identifies the Role of BZW2 in Promoting Endoplasmic Reticulum-Mitochondria Contact and Mitochondrial Metabolism.

Understanding the molecular functions of less-studied proteins is an important task of life science research. Despite reports of basic leucine zipper and W2 domain-containing protein 2 (BZW2) promoting cancer progression first emerging in 2017, little is known about its molecular function. Using a quantitative proteomic approach to identify its interacting proteins, we found that BZW2 interacts with both endoplasmic reticulum (ER) and mitochondrial proteins. We thus hypothesized that BZW2 localizes to and promotes the formation of ER-mitochondria contact sites and that such localization would promote calcium transport from ER to the mitochondria and promote ATP production. Indeed, we found that BZW2 localized to ER-mitochondria contact sites and that BZW2 knockdown decreased ER-mitochondria contact, mitochondrial calcium levels, and ATP production. These findings provide key insights into molecular functions of BZW2, the potential role of BZW2 in cancer progression, and highlight the utility of interactome data in understanding the function of less-studied proteins.

Removing Barriers to Emergency Medicine Point-of-Care Ultrasound: Illustrated by a Roadmap for Emergency Medicine Point-of-Care Ultrasound Expansion in India.

Point-of-care ultrasound (PoCUS) has a potentially vital role to play in emergency medicine (EM), whether it be in high-, medium-, or low-resourced settings. However, numerous barriers are present which impede EM PoCUS implementation nationally and globally: (i) lack of a national practice guideline or scope of practice for EM PoCUS, (ii) resistance from non-PoCUS users of ultrasound imaging (USI) and lack of awareness from those who undertake parallel or post-EM patient care, and (iii) heterogeneous pattern of resources available in different institutes and settings. When combined with the Indian Preconception and Prenatal Diagnostic Techniques (PCPNDT) Act, this has led to the majority of India's 1.4 billion citizens being unable to access EM PoCUS. In order to address these barriers (globally as well as with specific application to India), this article outlines the three core principles of EM PoCUS: (i) the remit of the EM PoCUS USI must be well defined a priori, (ii) the standard of EM PoCUS USI must be the same as that of non-PoCUS users of USI, and (iii) the imaging performed should align with subsequent clinical decision-making and resource availability. These principles are contextualized using an integrated PoCUS framework approach which is designed to provide a robust foundation for consolidation and expansion across different PoCUS specialisms and health-care settings. Thus, a range of mechanisms (from optimization of clinical practice through to PoCUS educational reform) are presented to address such barriers. For India, these are combined with specific mechanisms to address the PCPNDT Act, to provide the basis for influencing national legislation and instigating an addendum to the Act. By mapping to the recent Lancet Commission publication on transforming access to diagnostics, this provides a global and cross-discipline perspective for the recommendations.

Micro-CT Evaluation of Spontaneous Apexification of an Immature Tooth following Trauma.

This case reports on the micro-computerised tomography (CT) images of a periapical calcified dome following spontaneous apexification as a subsequence of trauma. An immature, maxillary central incisor was found to be non-vital one month following trauma. The tooth had minimal signs of structural damage; however, there was radiographic evidence of spontaneous apexification. The tooth suffered a second traumatic episode two years later and was decoronated to facilitate bone retention during osseous development. The patient presented with swelling and pain 36 months later, and the tooth root was extracted. The root was embedded in resin, and cross-sectional images were obtained using micro-CT. Analysis of the images provided insight into the structure of the calcified dome that formed following the first traumatic injury.

Complete Genome Sequences of the Type Strains Rouxiella badensis DSM 100043 and R. chamberiensis DSM 28324, Resolved Using Nanopore Long-Read Sequencing.

The complete genome sequences of Rouxiella badensis DSM 100043T and Rouxiella chamberiensis DSM 28324T were determined using Oxford Nanopore long-read sequencing and the Flye assembler. The former contains a circular chromosome of 4,964,479 bp and a circular plasmid of 116,582 bp; the latter contains a circular chromosome of 4,639,296 bp.

Genome comparison reveals that Halobacterium salinarum 63-R2 is the origin of the twin laboratory strains NRC-1 and R1.

The genome of Halobacterium strain 63-R2 was recently reported and provides the opportunity to resolve long-standing issues regarding the source of two widely used model strains of Halobacterium salinarum, NRC-1 and R1. Strain 63-R2 was isolated in 1934 from a salted buffalo hide (epithet "cutirubra"), along with another strain from a salted cow hide (91-R6T , epithet "salinaria," the type strain of Hbt. salinarum). Both strains belong to the same species according to genome-based taxonomy analysis (TYGS), with chromosome sequences showing 99.64% identity over 1.85 Mb. The chromosome of strain 63-R2 is 99.99% identical to the two laboratory strains NRC-1 and R1, with only five indels, excluding the mobilome. The two reported plasmids of strain 63-R2 share their architecture with plasmids of strain R1 (pHcu43/pHS4, 99.89% identity; pHcu235/pHS3, 100.0% identity). We detected and assembled additional plasmids using PacBio reads deposited at the SRA database, further corroborating that strain differences are minimal. One plasmid, pHcu190 (190,816 bp) corresponds to pHS1 (strain R1) but is even more similar in architecture to pNRC100 (strain NRC-1). Another plasmid, pHcu229, assembled partially and completed in silico (229,124 bp), shares most of its architecture with pHS2 (strain R1). In deviating regions, it corresponds to pNRC200 (strain NRC-1). Further architectural differences between the laboratory strain plasmids are not unique, but are present in strain 63-R2, which contains characteristics from both of them. Based on these observations, it is proposed that the early twentieth-century isolate 63-R2 is the immediate ancestor of the twin laboratory strains NRC-1 and R1.

An initial framework for use of ultrasound by speech and language therapists in the UK: Scope of practice, education and governance.

Background: There is growing evidence to support the use of ultrasound as a tool for the assessment and treatment of speech, voice and swallowing disorders across the Speech and Language Therapy profession. Research has shown that development of training competencies, engagement with employers and the professional body are vital to progressing ultrasound into practice. Methods: We present a framework to support translation of ultrasound into Speech and Language Therapy. The framework comprises three elements: (1) scope of practice, (2) education and competency and (3) governance. These elements align to provide a foundation for sustainable and high-quality ultrasound application across the profession. Results: Scope of practice includes the tissues to be imaged, the clinical and sonographic differentials and subsequent clinical decision-making. Defining this provides transformational clarity to Speech and Language Therapists, other imaging professionals and those designing care pathways. Education and competency are explicitly aligned with the scope of practice and include requisite training content and mechanisms for supervision/support from an appropriately trained individual in this area. Governance elements include legal, professional and insurance considerations. Quality assurance recommendations include data protection, storage of images, testing of ultrasound devices as well as continuous professional development and access to a second opinion. Conclusion: The framework provides an adaptable model for supporting expansion of ultrasound across a range of Speech and Language Therapy specialities. By taking an integrated approach, this multifaceted solution provides the foundation for those with speech, voice and swallowing disorders to benefit from advances in imaging-informed healthcare.

ICTV Virus Taxonomy Profile: Simuloviridae 2023.

The family Simuloviridae includes tailless icosahedral viruses with an internal lipid membrane. The capsid is constructed from two major capsid proteins, both with a single jelly-roll fold. The genome is a circular dsDNA molecule of 16-19 kb. All members infect halophilic archaea in the class Halobacteria (phylum Euryarchaeota) and are temperate viruses, their proviruses residing in host cells as extrachromosomal episomes. Once the lytic life cycle is triggered, production of virions causes cell lysis. This is a summary of the International Committee on Taxonomy of Viruses (ICTV) Report on the family Simuloviridae, which is available at ictv.global/report/simuloviridae.

ICTV Virus Taxonomy Profile: Sphaerolipoviridae 2023.

Members of the family Sphaerolipoviridae have non-enveloped tailless icosahedral virions with a protein-rich internal lipid membrane. The genome is a linear double-stranded DNA of about 30 kbp with inverted terminal repeats and terminal proteins. The capsid has a pseudo triangulation T=28 dextro symmetry and is built of two major capsid protein types. Spike complexes decorate fivefold vertices. Sphaerolipoviruses have a narrow host range and a lytic life cycle, infecting haloarchaea in the class Halobacteria (phylum Euryarchaeota). This is a summary of the International Committee on Taxonomy of Viruses (ICTV) Report on the family Sphaerolipoviridae, which is available at ictv.global/report/sphaerolipoviridae.

A proposed framework for point of care musculoskeletal ultrasound and ultrasound image-guided interventions by physiotherapists: scope of practice, education and governance.

BACKGROUND: The use of point of care ultrasound (PoCUS) in the management of musculoskeletal (MSK) disorders is a diverse area of PoCUS practice. Its use by clinicians, such as physiotherapists, can occur across a wide range of roles and care pathway configurations; however, professional, educational and regulatory uncertainties can leave clinicians, managers and patients at risk. MAIN BODY: A PoCUS framework approach (previously applied to support PoCUS consolidation and expansion) is used to frame these proposals. Central to this is the defining of (clinical and sonographic) scope of practice (ScoP). A number of indicative ScoPs are described to both (i) illustrate application of the principles and (ii) provide templates for ScoP derivations for individual services or clinicians. Image-guided MSK interventions are increasingly an aspect of MSK physiotherapy PoCUS. Given the utility of physiotherapists drawing upon their imaging to fully inform the selection (and performance) of such techniques, we present a rationale for competency in undertaking sonographic differentials as a pre-cursor to performing ultrasound image-guided MSK interventions. Alignment of ScoP with the relevant education and formal competency assessments are a cornerstone of the PoCUS framework approach; as such, key aspects of MSK PoCUS education and competency assessment are outlined. Strategies for addressing such requirements in healthcare settings where formal provision is not accessible, are also presented. Governance considerations are aligned with the regulatory environment, including those pertaining to professional guidance and insurance considerations. In addition, generic quality assurance elements are emphasised, as core aspects of high-quality service provision. Whilst the paper clarifies the situation for MSK physiotherapists using PoCUS in the UK, prompts are provided to support other professional groups working in MSK services in the United Kingdom (UK) and MSK physiotherapists/physical therapists in other countries-to facilitate their application of the principles. CONCLUSION: Acknowledging the breadth of MSK physiotherapy PoCUS practice, this paper draws upon a framework approach to provide integrated ScoP, education/competency and governance solutions, along with mechanisms for other professions working with MSK PoCUS-and physiotherapists/physical therapists outside of the UK-to consolidate and expand their practice.

Mach 18 flow velocimetry with 100-kHz KTV and PLEET in AEDC Tunnel 9.

Krypton Tagging Velocimetry (KTV) and Picosecond Laser Electronic Excitation Tagging (PLEET) velocimetry at a 100-kHz rate were demonstrated in Mach 18 flow conditions at the Arnold Engineering Development Center (AEDC) Tunnel 9 employing a burst-mode laser system and a custom optical parametric oscillator (OPO). The measured freestream flow velocities from both KTV and PLEET agreed well with the theoretical calculation. The increase in repetition rate provides better capability to perform time-resolved velocimetry measurements in hypersonic flow environments.

Role extension in advanced ultrasound practice: A framework approach and case study.

Introduction: Role extension into novel areas of ultrasound practice can be challenging for health care professionals. Expansion into existing areas of advanced practice typically occurs using established processes and accredited training; however, in areas where there is no formal training, there can be a lack of support for how to develop new and progressive clinical roles. Topic Description: This article presents how the use of a framework approach for establishing areas of advanced practice can support individuals and departments with safely and successfully developing new roles in ultrasound. The authors illustrate this via the example of a gastrointestinal ultrasound role, developed in an NHS department. Discussion: The framework approach comprises three elements, each interdependent upon and inform each other: (A) Scope of practice, (B) Education and competency and (C) Governance. (A) Defines (and communicates) the role extension and area(s) of subsequent ultrasound imaging, interpretation and reporting. By identifying the why, how and what is required this informs (B) the education and assessment of competency for those taking on new roles or areas of expertise. (C) Is informed by (A) and is an ongoing process of quality assurance to safeguard high standards in clinical care. In supporting role extension, this approach can facilitate new workforce configurations, skill expansion and enable increasing service demands to be met. Summary: By defining and aligning the components of scope of practice, education/competency and governance, role development in ultrasound can be initiated and sustained. Role extension utilising this approach brings benefits for patients, clinicians and departments.

The ability of physiotherapists to identify psychosocial factors in patients with musculoskeletal pain: A scoping review.

BACKGROUND: It is not known how well physiotherapists identify psychosocial factors in people with musculoskeletal pain, when using clinical judgement. The purpose of this scoping review was to examine the research related to physiotherapist ability in identifying psychosocial factors and to subsequently identify gaps in the literature to help direct future research. DATA SOURCES: Searches using relevant key words, were conducted of Medline, Cinahl, the Cochrane Library, PEDro, PubMed, Scopus and Google Scholar. All primary quantitative and qualitative research from the year 2000 onwards, which met the search criteria, were included. DATA EXTRACTION AND SYNTHESIS: A data extraction tool was used to tabulate data regarding demographics, study design and key findings of the included papers. The Mixed Methods Appraisals Tool (MMAT) was utilised to help examine the quality of included studies. RESULTS: Overall, the quality of the included studies was moderate. The total number of studies which met the inclusion criteria was relatively small (n = 20). The most common method for determining ability was comparison of physiotherapist estimations with validated screening tools or questionnaires. Physiotherapist estimates of psychosocial factors were poor and in the qualitative research, the lack of clinician confidence in psychosocial assessment was evident. CONCLUSION: The available research suggests that physiotherapists lack confidence and ability in identifying psychosocial factors. More rigorous, mixed-methods research is warranted to capture the complexity of the research question.