Aspiration removal of orbitofrontal cortex disrupts cholinergic fibers of passage to anterior cingulate cortex in rhesus macaques.

The study of anthropoid nonhuman primates has provided valuable insights into frontal cortex function in humans, as these primates share similar frontal anatomical subdivisions (Murray et al. 2011). Causal manipulation studies have been instrumental in advancing our understanding of this area. One puzzling finding is that macaques with bilateral aspiration removals of orbitofrontal cortex (OFC) are impaired on tests of cognitive flexibility and emotion regulation, whereas those with bilateral excitotoxic lesions of OFC are not (Rudebeck et al. 2013). This discrepancy is attributed to the inadvertent disruption of fibers of passage by aspiration lesions but not by excitotoxic lesions. Which fibers of passage are responsible for the impairments observed? One candidate is cholinergic fibers originating in the nucleus basalis magnocellularis (NBM) and passing nearby or through OFC on their way to other frontal cortex regions (Kitt et al. 1987). To investigate this possibility, we performed unilateral aspiration lesions of OFC in three macaques, and then compared cholinergic innervation of the anterior cingulate cortex (ACC) between hemispheres. Histological assessment revealed diminished cholinergic innervation in the ACC of hemispheres with OFC lesions relative to intact hemispheres. This finding indicates that aspiration lesions of the OFC disrupt cholinergic fibers of passage, and suggests the possibility that loss of cholinergic inputs to ACC contributes to the impairments in cognitive flexibility and emotion regulation observed after aspiration but not excitotoxic lesions of OFC.

Facilitators of, barriers to, and preferences for e-mental health interventions for depression and anxiety in men: Metasynthesis and recommendations.

BACKGROUND: Little is known about men's use of online mental health (eMH) interventions and factors that promote their engagement or attrition. We aimed to synthesise the qualitative literature on men's preferences for, attitudes towards, and experiences of using eMH interventions for depression and anxiety, and develop recommendations from the findings. METHOD: Systematic searches were conducted (Jan 2000-Oct 2020) in six databases; study quality was assessed using Qualsyst with a minimum total of 0.55 required for inclusion. Extracted data were synthesised using meta-aggregation. RESULTS: Eight studies met inclusion criteria and three synthesised findings were generated. (1) Facilitators of men's eMH use: finding apps and technology motivating and convenient, support and encouragement from important others, and interventions allowing men to take action, gain control over their mental health, and resulting in positive outcomes; (2) Barriers to men's eMH use: lack of free time, predicted or experienced lack of benefit from use, and technical difficulties; (3) What men want in eMH: personalised, tailored, relevant interventions that are bright and easy to use, with information presented in multiple formats, psychoeducation, exercises, self-monitoring, information on further resources, and the option of clinician involvement, without any repetitive questioning, boring tools, or negative feedback. LIMITATIONS: All included studies were conducted in high income, 'Western' countries; most data related to experiences of using an existing eMH intervention within a trial, rather than in 'real world' settings where eMH acceptability is generally lower and experiences may differ. CONCLUSIONS: Practice, research, and policy recommendations are presented.

Articular Surface Involvement With Retrograde Headless Screw Fixation of the Proximal Phalanx.

PURPOSE: Intramedullary (IM) screw fixation of proximal phalanx (P1) fractures is a treatment option increasing in popularity. This study aimed to quantify the articular surface loss after retrograde screw insertion and to determine the range of motion (ROM) of the proximal interphalangeal (PIP) joint while the defect in the P1 head is engaged with the base of the middle phalanx (P2). METHODS: Twelve fresh frozen cadaver hand specimens were analyzed for prefixation ROM of the PIP joint. A retrograde screw was placed using a percutaneous technique under fluoroscopic guidance. Following screw insertion, specimens were dissected to determine size of the extensor mechanism defect, evaluate the lateral bands with passive ROM of the PIP joint, and determine the angle at which the dorsal aspect of P2 ceases to engage with the defect and the amount of articular surface loss. The percentage of articular surface loss was calculated using a digital image software program. RESULTS: The angle at which P2 ceased to engage with the articular surface defect was an average of 36.8° of PIP joint flexion. In full PIP joint flexion, the average extensor mechanism defect was 8.8%. The average total articular surface loss was 4.4% across all digits. CONCLUSION: Percutaneous retrograde P1 intramedullary screw fixation results in minimal damage to the extensor mechanism and articular surface. The arc during which the defect in the head of P1 engages the base of the P2 is almost entirely outside the functional ROM of the PIP joint. CLINICAL RELEVANCE: Quantifying the amount of articular surface loss through the P1 head and extensor apparatus damage in IM screw fixation can inform surgeons of the consequences of this technique. This study supports the use of a retrograde intramedullary screw as a safe option for fixation of P1 fractures.

Excretory-secretory products from adult helminth Nippostrongylus brasiliensis have in vitro bactericidal activity.

Introduction. Intestinal helminths and microbiota share the same anatomical niche during infection and are likely to interact either directly or indirectly. Whether intestinal helminths employ bactericidal strategies that influence their microbial environment is not completely understood.Hypothesis. In the present study, the hypothesis that the adult hookworm Nippostrongylus brasiliensis produces molecules that impair bacterial growth in vitro, is tested.Aim. To investigate the in vitro bactericidal activity of Nippostrongylus brasiliensis against commensal and pathogenic bacteria.Methodology. The bactericidal effect of somatic extract and excretory-secretory products of adult Nippostrongylus brasiliensis on Gram-positive (Staphylococcus aureus) and Gram-negative (Escherichia coli, Salmonella enterica serovar Typhimurium, and Klebsiella pneumoniae) bacteria was assessed using growth assays. Minimum inhibitory concentration and minimum bactericidal concentration assays were performed using excretory-secretory products released from the pathogen.Results. Broad-spectrum in vitro bactericidal activity in excretory-secretory products, but not somatic extract of adult Nippostrongylus brasiliensis was detected. The bactericidal activity of excretory-secretory products was concentration-dependent, maintained after heat treatment, and preserved after repeated freezing and thawing.Conclusion. The results of this study demonstrate that helminths such as Nippostrongylus brasiliensis release molecules via their excretory-secretory pathway that have broad-spectrum bactericidal activity. The mechanisms responsible for this bactericidal activity remain to be determined and further studies aimed at isolating and identifying active bactericidal molecules are needed.

Identification of proteins that bind extracellular microRNAs secreted by the parasitic nematode Trichinella spiralis.

Small non-coding RNAs such as microRNAs (miRNAs) are conserved across eukaryotes and play key roles in regulating gene expression. In many organisms, miRNAs are also secreted from cells, often encased within vesicles such as exosomes, and sometimes extravesicular. The mechanisms of miRNA secretion, how they are stabilised outside of cells and their functional importance are poorly understood. Recently, we characterised the parasitic nematode Trichinella spiralis as a model to study miRNA secretion. T. spiralis muscle-stage larvae (MSL) secrete abundant miRNAs which are largely extravesicular. Here, we investigated how T. spiralis miRNAs might remain stable outside of cells. Using proteomics, we identified two RNA binding proteins secreted by T. spiralis larvae and characterised their RNA binding properties. One, a homologue of the known RNA binding protein KSRP, binds miRNA in a selective and sequence-specific fashion. Another protein, which is likely a novel RNA binding protein, binds to miRNA without exhibiting sequence specificity. Our results suggest a possible mechanism for miRNA secretion by T. spiralis and may have relevance for understanding the biology of extracellular miRNA more widely.

Familial and Bullying Victimisation: The Impact of Early Adversity Within the Home and Peer Settings on Late Adolescence and Adult Psychopathology.

Background: Awareness of adverse childhood experiences and their impact on adult psychopathology primarily focuses on adversities within the home. There is limited insight into the impact of adversities across peer environments. Objective: This study investigates 19 items related to adverse experiences across the home, school and peer environments and their relationship to 12-month and lifetime psychopathology. Data: Secondary analysis of the Ulster University Student Well-being Study. The dataset included completed responses across all selected variables for 729 participants. Method and Results: Latent profile analysis identified a low adversity profile, bullying adversity profile and higher prevalence adversity profile. Regression analysis of the three profiles and demographics variables indicated their impact on adult psychopathology lifetime and 12-month prevalence rates. Conclusion: Schools and HE institutions should acknowledge the impact of childhood adversities. In doing so, it is important to consider the deeper impact of bullying due to its links with psychopathology across the lifespan. Educational institutions should take appropriate steps to mitigate continued exposure as students' progress through the education system.

Exploring the integration of diagnostic musculoskeletal ultrasound imaging into clinical practice by physical therapists.

BACKGROUND: Musculoskeletal ultrasound (MSK-US) use for diagnostic purposes is expanding in physical therapy practice. Identifying and describing physical therapy-specific approaches to incorporating MSK-US into the evaluation process is needed. Musculoskeletal ultrasound extends the physical exam to allow clinicians to visualize anatomy and pathophysiology both statically and dynamically. Purpose: To document 1) weekly use of diagnostic MSK-US; and 2) clinical reasoning approach used in challenging patient cases by physical therapists (PTs) registered by Inteleos in musculoskeletal sonography (RMSK-certified). METHODS: Longitudinal, observational, cohort study using mixed methods for data collection and analysis. All 23 currently RMSK-certified PTs using MSK-US in clinical practice across the United States were contacted, and 16 participated. Data were collected using an online survey created with the Research Electronic Data Capture System. Participants documented MSK-US clinical use and significant cases using weekly, reflective, online journals for three months. Demographic data were summarized using descriptive statistics. Case data were analyzed thematically. RESULTS: Participating RMSK-certified PTs performed 1110 MSK-US examinations over 110 weeks. Clinicians averaged 7 (range 1-25) MSK-US examinations weekly, representing 28% of an average caseload. Examinations contributed significant anatomical/ pathological information 100% of the time. The most common joints scanned were the knee (n = 281), shoulder (n = 254), and wrist (n = 228). Case data revealed three themes: 1) augmenting the clinical evaluation to extend or narrow a diagnosis; 2) outcomes guiding action; and 3) lessons learned from clinical findings. CONCLUSION: RMSK-certified PTs regularly used MSK-US to validate and refine their clinical diagnoses and treatment. Ultrasound imaging directly influenced patient care by informing the diagnostic process, guiding treatment, and appropriately identifying referrals.

Mechanical testing of frontal plane adaptability of commercially available prosthetic feet.

Introduction: Prosthetic feet have limited adaptability in the frontal plane. Research shows walking on uneven terrain is difficult for many prosthesis users. A new prosthetic foot, the META Arc, was designed with a polycentric ankle joint that allows relatively free movement in the frontal plane to address this limitation. Previous simulations of the polycentric ankle mechanism found potential benefits such as reduced lateral movement of a proximal mass during forward progress and reduced forces being transferred upward from the ground through the foot. Methods: Standard mechanical testing protocols were used to evaluate the Meta Arc prosthetic foot's performance and six comparable feet commercially available. Results: The results found the META Arc prosthetic foot had increased frontal plane adaptability as well as reduced lateral forces, and reduced inversion eversion moment compared to the six comparison feet on 10-degree cross-slope test conditions. All included prosthetic feet had similar results for the percent of energy return and dynamic force in the sagittal plane. Conclusions: These results suggest the inclusion of the polycentric ankle within the META Arc foot will provide more stability without sacrificing forward walking performance.

Temporal profiling of CD4 T-cell activation and differentiation upon SARS-CoV-2 spike protein immunisation

CD4 T-cells require T-cell receptor (TCR) signalling for their activation and differentiation. Foxp3+ regulatory T-cells (Treg) are dependent on TCR signals for their differentiation and suppressive function. However, it is not fully known how TCR signalling controls the differentiation of polyclonal CD4 T-cells upon antigen recognition at the single-cell level in vivo. In this study, using Nr4a3-Tocky (Timer-of-cell-kinetics-and-activity), which analyses temporal changes of antigen-reactive T-cells following TCR signalling, we investigated T-cell response to Spike protein fragments (S1a, S1b, S2a, and S2b) upon immunisation. We show that S1a and S2a induced the differentiation of PD1hiCXCR5+ T follicular helper (Tfh) cells, which is related to CD4 T-cell immunogenicity. In contrast, S1b induced CD25hiGITRhiPD-1int Treg, which intermittently received TCR signalling. Using Foxp3-Tocky, which analyses Foxp3 transcriptional dynamics, the S1b-reactive Treg sustained Foxp3 transcription over time, which is a hallmark of activated Treg. Foxp3 fate-mapping showed that the S1b-reactive Treg were derived not from pre-existing thymic Treg, suggesting Foxp3 induction in non-Treg cells. Thus, the current study reveals temporally dynamic differentiation of CD4 T-cells and Treg upon immunisation in the polyclonal TCR repertoire.

Differential Regulation of Allergic Airway Inflammation by Acetylcholine.

Acetylcholine (ACh) from neuronal and non-neuronal sources plays an important role in the regulation of immune responses and is associated with the development of several disease pathologies. We have previously demonstrated that group 2 innate lymphoid cell (ILC2)-derived ACh is required for optimal type 2 responses to parasitic infection and therefore sought to determine whether this also plays a role in allergic inflammation. RoraCre+ChatLoxP mice (in which ILC2s cannot synthesize ACh) were exposed to an allergenic extract of the fungus Alternaria alternata, and immune responses in the airways and lung tissues were analyzed. Airway neutrophilia and expression of the neutrophil chemoattractants CXCL1 and CXCL2 were enhanced 24 h after exposure, suggesting that ILC2-derived ACh plays a role in limiting excessive pulmonary neutrophilic inflammation. The effect of non-selective depletion of ACh was examined by intranasal administration of a stable parasite-secreted acetylcholinesterase. Depletion of airway ACh in this manner resulted in a more profound enhancement of neutrophilia and chemokine expression, suggesting multiple cellular sources for the release of ACh. In contrast, depletion of ACh inhibited Alternaria-induced activation of ILC2s, suppressing the expression of IL-5, IL-13, and subsequent eosinophilia. Depletion of ACh reduced macrophages with an alternatively activated M2 phenotype and an increase in M1 macrophage marker expression. These data suggest that ACh regulates allergic airway inflammation in several ways, enhancing ILC2-driven eosinophilia but suppressing neutrophilia through reduced chemokine expression.

Aquifer heterogeneity controls to quality monitoring network performance for the protection of groundwater production wells.

A groundwater monitoring network surrounding a pumping well (such as a public water supply) allows for early contaminant detection and mitigation where possible contaminant source locations are often unknown. This numerical study investigates how the contaminant detection probability of a hypothetical sentinel-well monitoring network consisting of one to four monitoring wells is affected by aquifer spatial heterogeneity and dispersion characteristics, where the contaminant source location is randomized. This is achieved through a stochastic framework using a Monte Carlo approach. A single production well is considered that results in converging non-uniform flow close to the well. Optimal network arrangements are obtained by maximizing a weighted risk function that considers true and false positive detection rates, sampling frequency, early detection, and contaminant travel time uncertainty. Aquifer dispersivity is found to be the dominant parameter for the quantification of network performance. For the range of parameters considered, a single monitoring well screening the full aquifer thickness is expected to correctly and timely identify at least 12% of all incidents resulting in contaminants reaching the production well. This proportion increases to a global maximum of 96% for a network consisting of four wells and very dispersive transport conditions. Irrespective of network size and sampling frequency, more dispersive transport conditions result in higher detection rates. Increasing aquifer heterogeneity and decreasing aquifer spatial continuity also lead to higher detection rates, though these effects are diminished for networks of 3 or more wells. Statistical anisotropy has no effect on the network performance. Earlier detection, which is critical for remedial action and supply safety, comes with a significant cost in terms of detection rate, and should be carefully considered when a monitoring network is being designed.

Observation of photo-induced plasmon-phonon coupling in PbTe via ultrafast x-ray scattering.

We report the observation of photo-induced plasmon-phonon coupled modes in the group IV-VI semiconductor PbTe using ultrafast x-ray diffuse scattering at the Linac Coherent Light Source. We measure the near-zone-center excited-state dispersion of the heavily screened longitudinal optical (LO) phonon branch as extracted from differential changes in x-ray diffuse scattering intensity following above bandgap photoexcitation. We suggest that upon photoexcitation, the LO phonon-plasmon coupled (LOPC) modes themselves become coupled to longitudinal acoustic modes that drive electron band shifts via acoustic deformation potentials and possibly to low-energy single-particle excitations within the plasma and that these couplings give rise to displacement-correlations that oscillate in time with a period given effectively by the heavily screened LOPC frequency.

Systemic and vascular inflammation in experimental allergic asthma.

Allergic asthma and atherosclerosis are inflammatory diseases characterized by similar sets of circulating inflammatory cells, in addition to mast cells in the airway and vessel wall. Animal models and human studies provide evidence of a potential interaction between the two apparently unrelated diseases. The main objective of this study was to determine whether experimental allergic asthma is accompanied by inflammatory responses, measured as the activation of the vasculature and the presence of immune cells in the perivascular adipose tissue. For this purpose, male Dunkin Hartley guinea pigs weighing 250 - 300 g were sensitized twice with 10 µg ovalbumin dissolved in aluminium hydroxide (Al(OH)3). Allergen inhalation was performed 10 days after the second immunization and continued 5 days a week for 2 months. After that period, T cell and macrophage content was measured by flow cytometry. The aortic expression of inflammatory markers was studied by real-time PCR. The number of T cells in the peripheral blood was significantly greater in the allergic group in comparison to the sham group. We did not find any significant differences in the leukocyte content of the perivascular adipose tissue between the groups. Nor did we identify significant changes in the expression of inflammatory markers (tumor necrosis factor, monocyte chemoattractant protein-1) and adhesion molecules (intercellular adhesion molecules and vascular cell adhesion molecules) in the aorta. Interestingly, we observed a significantly decreased expression of the endothelial nitric oxide synthase (eNOS) mRNA in the aortic vessel of the allergic group compared to the sham group.

The presence and potential impact of psychological safety in the healthcare setting: an evidence synthesis.

INTRODUCTION: Psychological safety is the shared belief that the team is safe for interpersonal risk taking. Its presence improves innovation and error prevention. This evidence synthesis had 3 objectives: explore the current literature regarding psychological safety, identify methods used in its assessment and investigate for evidence of consequences of a psychologically safe environment. METHODS: We searched multiple trial registries through December 2018. All studies addressing psychological safety within healthcare workers were included and reviewed for methodological limitations. A thematic analysis approach explored the presence of psychological safety. Content analysis was utilised to evaluate potential consequences. RESULTS: We included 62 papers from 19 countries. The thematic analysis demonstrated high and low levels of psychological safety both at the individual level in study participants and across the studies themselves. There was heterogeneity in responses across all studies, limiting generalisable conclusions about the overall presence of psychological safety. A wide range of methods were used. Twenty-five used qualitative methodology, predominantly semi-structured interviews. Thirty quantitative or mixed method studies used surveys. Ten studies inferred that low psychological safety negatively impacted patient safety. Nine demonstrated a significant relationship between psychological safety and team outcomes. The thematic analysis allowed the development of concepts beyond the content of the original studies. This analytical process provided a wealth of information regarding facilitators and barriers to psychological safety and the development of a model demonstrating the influence of situational context. DISCUSSION: This evidence synthesis highlights that whilst there is a positive and demonstrable presence of psychological safety within healthcare workers worldwide, there is room for improvement. The variability in methods used demonstrates scope to harmonise this. We draw attention to potential consequences of both high and low psychological safety. We provide novel information about the influence of situational context on an individual's psychological safety and offer more detail about the facilitators and barriers to psychological safety than seen in previous reviews. There is a risk of participation bias - centres involved in safety research may be more aligned to these ideals. The data in this synthesis are useful for institutions looking to improve psychological safety by providing a framework from which modifiable factors can be identified.

Ancient DNA, lipid biomarkers and palaeoecological evidence reveals construction and life on early medieval lake settlements.

Direct evidence of ancient human occupation is typically established through archaeological excavation. Excavations are costly and destructive, and practically impossible in some lake and wetland environments. We present here an alternative approach, providing direct evidence from lake sediments using DNA metabarcoding, steroid lipid biomarkers (bile acids) and from traditional environmental analyses. Applied to an early Medieval Celtic settlement in Ireland (a crannog) this approach provides a site chronology and direct evidence of human occupation, crops, animal farming and on-site slaughtering. This is the first independently-dated, continuous molecular archive of human activity from an archeological site, demonstrating a link between animal husbandry, food resources, island use. These sites are under threat but are impossible to preserve in-situ so this approach can be used, with or without excavation, to produce a robust and full site chronology and provide direct evidence of occupation, the use of plants and animals, and activities such as butchery.

Eosinophilic pneumonia caused by cefepime: A case report and review.

Eosinophilic pneumonia (EP) is characterized by accumulation of eosinophils in the lungs and has been associated with several medications, including antimicrobials. Cefepime is a commonly used broad-spectrum antimicrobial agent for the treatment of nosocomial infections but to date has not been associated with EP. We report the first documented case of EP secondary to cefepime for the treatment of pneumonia. The patient's peripheral eosinophilia and leukocytosis resolved promptly after discontinuation of cefepime and initiation of steroid treatment.

Component Parts of Bacteriophage Virions Accurately Defined by a Machine-Learning Approach Built on Evolutionary Features.

Antimicrobial resistance (AMR) continues to evolve as a major threat to human health, and new strategies are required for the treatment of AMR infections. Bacteriophages (phages) that kill bacterial pathogens are being identified for use in phage therapies, with the intention to apply these bactericidal viruses directly into the infection sites in bespoke phage cocktails. Despite the great unsampled phage diversity for this purpose, an issue hampering the roll out of phage therapy is the poor quality annotation of many of the phage genomes, particularly for those from infrequently sampled environmental sources. We developed a computational tool called STEP3 to use the "evolutionary features" that can be recognized in genome sequences of diverse phages. These features, when integrated into an ensemble framework, achieved a stable and robust prediction performance when benchmarked against other prediction tools using phages from diverse sources. Validation of the prediction accuracy of STEP3 was conducted with high-resolution mass spectrometry analysis of two novel phages, isolated from a watercourse in the Southern Hemisphere. STEP3 provides a robust computational approach to distinguish specific and universal features in phages to improve the quality of phage cocktails and is available for use at http://step3.erc.monash.edu/. IMPORTANCE In response to the global problem of antimicrobial resistance, there are moves to use bacteriophages (phages) as therapeutic agents. Selecting which phages will be effective therapeutics relies on interpreting features contributing to shelf-life and applicability to diagnosed infections. However, the protein components of the phage virions that dictate these properties vary so much in sequence that best estimates suggest failure to recognize up to 90% of them. We have utilized this diversity in evolutionary features as an advantage, to apply machine learning for prediction accuracy for diverse components in phage virions. We benchmark this new tool showing the accurate recognition and evaluation of phage component parts using genome sequence data of phages from undersampled environments, where the richest diversity of phage still lies.

MicroRNA-142 Critically Regulates Group 2 Innate Lymphoid Cell Homeostasis and Function.

Innate lymphoid cells are central to the regulation of immunity at mucosal barrier sites, with group 2 innate lymphoid cells (ILC2s) being particularly important in type 2 immunity. In this study, we demonstrate that microRNA(miR)-142 plays a critical, cell-intrinsic role in the homeostasis and function of ILC2s. Mice deficient for miR-142 expression demonstrate an ILC2 progenitor-biased development in the bone marrow, and along with peripheral ILC2s at mucosal sites, these cells display a greatly altered phenotype based on surface marker expression. ILC2 proliferative and effector functions are severely dysfunctional following Nippostrongylus brasiliensis infection, revealing a critical role for miR-142 isoforms in ILC2-mediated immune responses. Mechanistically, Socs1 and Gfi1 expression are regulated by miR-142 isoforms in ILC2s, impacting ILC2 phenotypes as well as the proliferative and effector capacity of these cells. The identification of these novel pathways opens potential new avenues to modulate ILC2-dependent immune functions.