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Abstract Background Many medical trainees, prior to achieving specialist status, are required to complete a mandatory research project, the usefulness of which has been debated. The aim of this study was to gain an in-depth understanding of trainees’ experiences and satisfaction of conducting such research projects in Australia. Methods A qualitative descriptive approach was used. Semi-structured interviews with trainees were undertaken between May 2021 and June 2022. Australian medical trainees who had completed a research project as part of specialty training within the past five years were invited to participate. The purposive sample was drawn from participants in a survey on the same topic who had indicated interest in participating in an interview. Interviews explored trainees’ overall experience of and satisfaction with conducting research projects, as well as their perceptions of research training, support, barriers, enablers, and perceived benefits. Interviews were transcribed verbatim and thematically analysed. Results Sixteen medical doctors from seven medical colleges were interviewed. Trainee experience and satisfaction was highly variable between participants and was shaped by four factors: 1) trainees entered their specialty training with their own perspectives on the value and purpose of the research project, informed by their previous experiences with research and perceived importance of research in their planned career path; 2) in conducting the project, enablers including protected time, supervisor support and institutional structures, were vital to shaping their experience; 3) trainees’ access to these enablers was variable, mediated by a combination of luck, and the trainees’ own drive and research skill; and 4) project outcomes, in terms of research merit, learning, career benefits and impacts on patient care. Conclusions Trainee experiences of doing research were mixed, with positive experiences often attributed to chance rather than an intentionally structured learning experience. We believe alternatives to mandatory trainee research projects must be explored, including recognising other forms of research learning activities, and directing scarce resources to supporting the few trainees who plan to pursue clinician researcher careers.

Abstract Estrogens regulate numerous physiological and pathological processes, including wide-ranging effects in wound healing. The effects of estrogens are mediated through multiple estrogen receptors (ERs), including the classical nuclear ERs (ERα and ER $${\upbeta }$$ β ), that typically regulate gene expression, and the 7-transmembrane G protein-coupled estrogen receptor (GPER), that predominantly mediates rapid “non-genomic” signaling. Estrogen modulates the expression of various genes involved in epidermal function and regeneration, inflammation, matrix production, and protease inhibition, all critical to wound healing. Our previous work demonstrated improved myocutaneous wound healing in female mice compared to male mice. In the current study, we employed male and female GPER knockout mice to investigate the role of this estrogen receptor in wound revascularization and tissue viability. Using a murine myocutaneous flap model of graded ischemia, we measured real-time flap perfusion via laser speckle perfusion imaging. We conducted histologic and immunohistochemical analyses to assess skin and muscle viability, microvascular density and vessel morphology. Our results demonstrate that GPER is crucial in wound healing, mediating effects that are both dependent and independent of sex. Lack of GPER expression is associated with increased skin necrosis, reduced flap perfusion and altered vessel morphology. These findings contribute to understanding GPER signaling in wound healing and suggest possible therapeutic opportunities by targeting GPER.

Abstract Vitamin K antagonists (VKA) remain the only option of anticoagulation for people with mechanical valve replacement and due to their wider availability and lower acquisition costs, VKA’s remain widely used in low- and middle-income countries. It has been suggested that prolonged use of VKAs can increase the development of vascular and valvular calcification, though this effect has not been examined in larger randomized prospective trials. This investigator-initiated multicenter, prospective, randomized, open-label interventional trial randomized patients with baseline coronary or valvular calcification and an indication for prolonged oral anticoagulation therapy to Marcumar or Rivaroxaban. Patients were followed-up through repeat coronary computed tomographies to measure the progression of coronary and valvular calcification for up to 24 months. 192 patients were randomized between 2013 and 2018 to receive either Rivaroxaban or Marcumar and followed for up to 24 months. Coronary calcification significantly increased over time although there was no significant difference in progression between the groups after 12 and 24 months as measured by the Agatston score [360.7 (90.2; 1075.3) vs 380.4 (136.4; 1546.9) p = 0.69], the volume score [295.8 (93.0; 995.3) vs 335.5 (128.7; 1316.9) p = 0.95] and the mass score [58.5 (15.9; 172.0) vs 71.1 (24.8; 257.3) p = 0.5]. Dephosphorylated, uncarboxylated matrix Gla Protein (Dp-ucMGP) significantly decreased in the VKA group [Δ dp-uc MGP – 95.2 (− 554.1; 156.0) vs 231.3 (− 59.7; 388.1) p

Abstract Helminth infections lead to an overdispersion of the parasites in humans as well as in animals. We asked whether early immune responses against migrating Ascaris larvae are responsible for the unequal distribution of worms in natural host populations and thus investigated a susceptible versus a resistant mouse strain. In mice, the roundworm larvae develop until the lung stage and thus early anti-Ascaris immune responses against the migrating larvae in the liver and lung can be deciphered. Our data show that susceptible C57BL/6 mice respond to Ascaris larval migration significantly stronger compared to resistant CBA mice and the anti-parasite reactivity is associated with pathology. Increased eosinophil recruitment was detected in the liver and lungs, but also in the spleen and peritoneal cavity of susceptible mice on day 8 post infection compared to resistant mice. In serum, eosinophil peroxidase levels were significantly higher only in the susceptible mice, indicating functional activity of the recruited eosinophils. This effect was associated with an increased IL-5/IL-13 production by innate lymphoid cells and CD4+ T cells and a pronounced type 2 macrophage polarization in the lungs of susceptible mice. Furthermore, a comparison of wildtype BALB/c and eosinophil-deficient dblGATA-1 BALB/c mice showed that eosinophils were not essential for the early control of migrating Ascaris larvae. In conclusion, in primary infection, a strong local and systemic type 2 immune response during hepato-tracheal helminth larval migration is associated with pathology rather than protection.

Abstract Natural killer (NK) cells play a key role in defense against Salmonella infections during the early phase of infection. Our previous work showed that the excretory/secretory products of Ascaris suum repressed NK activity in vitro. Here, we asked if NK cell functionality was influenced in domestic pigs during coinfection with Ascaris and Salmonella enterica serotype Typhimurium. Ascaris coinfection completely abolished the IL-12 and IL-18 driven elevation of IFN-γ production seen in CD16 + CD8α + perforin + NK cells of Salmonella single-infected pigs. Furthermore, Ascaris coinfection prohibited the Salmonella-driven rise in NK perforin levels and CD107a surface expression. In line with impaired effector functions, NK cells from Ascaris-single and coinfected pigs displayed elevated expression of the inhibitory KLRA1 and NKG2A receptors genes, contrasting with the higher expression of the activating NKp46 and NKp30 receptors in NK cells during Salmonella single infection. These differences were accompanied by the highly significant upregulation of T-bet protein expression in NK cells from Ascaris-single and Ascaris/Salmonella coinfected pigs. Together, our data strongly indicate a profound repression of NK functionality by an Ascaris infection which may hinder infected individuals from adequately responding to a concurrent bacterial infection.

Abstract Without the protective shielding of Earth’s atmosphere, astronauts face higher doses of ionizing radiation in space, causing serious health concerns. Highly charged and high energy (HZE) particles are particularly effective in causing complex and difficult-to-repair DNA double-strand breaks compared to low linear energy transfer. Additionally, chronic cortisol exposure during spaceflight raises further concerns, although its specific impact on DNA damage and repair remains unknown. This study explorers the effect of different radiation qualities (photons, protons, carbon, and iron ions) on the DNA damage and repair of cortisol-conditioned primary human dermal fibroblasts. Besides, we introduce a new measure, the Foci-Integrated Damage Complexity Score (FIDCS), to assess DNA damage complexity by analyzing focus area and fluorescent intensity. Our results show that the FIDCS captured the DNA damage induced by different radiation qualities better than counting the number of foci, as traditionally done. Besides, using this measure, we were able to identify differences in DNA damage between cortisol-exposed cells and controls. This suggests that, besides measuring the total number of foci, considering the complexity of the DNA damage by means of the FIDCS can provide additional and, in our case, improved information when comparing different radiation qualities.

BackgroundParticularly in rural regions, factors such as lower physician density and long travel distances complicate adequate outpatient care. However, urban regions can also be affected by deficits in care, for example, long waiting times. One model of care intending to improve the situation is the implementation of video consultations. The study protocol presents the methodology of the research project titled “Preference-based implementation of the video consultation in urban and rural regions” funded by the German Federal Joint Committee (funding number 01VSF20011). ObjectiveThis study aims to identify existing barriers to the use of video consultation and the preferences of insured individuals and physicians as well as psychotherapists in order to optimize its design and thus increase acceptance and use of video consultations in urban and rural regions. MethodsBuilt on a mixed methods approach, this study first assesses the status quo of video consultation use through claims data analysis and carries out a systematic literature review on barriers and promoting factors for the use of video consultations. Based on this preliminary work, focus groups are conducted in order to prepare surveys with insureds as well as physicians and psychotherapists in the second study phase. The central element of the survey is the implementation of discrete choice experiments to elicit relevant preferences of (potential) user groups and service providers. The summarized findings are discussed in a stakeholder workshop and translated into health policy recommendations. ResultsThe methodological approach used in this study is the focus of this paper. The study is still ongoing and will continue until March 2024. The first study phase has already been completed, in which preliminary work has been done on potential applications and hurdles for the use of video consultations. Currently, the survey is being conducted and analyses are being prepared. ConclusionsThis study is intended to develop a targeted strategy for health policy makers based on actual preferences and perceived obstacles to the use of video consultations. The results of this study will contribute to further user-oriented development of the implementation of video consultations in German statutory health insurance. Furthermore, the iterative and mixed methods approach used in this study protocol is also suitable for a variety of other research projects. International Registered Report Identifier (IRRID)DERR1-10.2196/50932

AimsFear of physical activity (PA) is discussed as a barrier to regular exercise in patients with heart failure (HF), but HF-specific theoretical concepts are lacking. This study examined associations of fear of PA, heart-focused anxiety and trait anxiety with clinical characteristics and self-reported PA in outpatients with chronic HF. It was also investigated whether personality-related coping styles for dealing with health threats impact fear of PA via symptom perception.Methods and resultsThis cross-sectional study enrolled 185 HF outpatients from five hospitals (mean age 62 ± 11 years, mean ejection fraction 36.0 ± 12%, 24% women). Avoidance of PA, sports/exercise participation (yes/no) and the psychological characteristics were assessed by self-reports. Fear of PA was assessed by the Fear of Activity in Situations-Heart Failure (FActS-HF15) questionnaire. In multivariable regression analyses higher NYHA class (b = 0.26, p = 0.036) and a higher number of HF drugs including antidepressants (b = 0.25, p = 0.017) were independently associated with higher fear of PA, but not with heart-focused fear and trait anxiety. Of the three anxiety scores only increased fear of PA was independently associated with more avoidance behavior regarding PA (b = 0.45, SE = 0.06, p < 0.001) and with increased odds of no sports/exercise participation (OR = 1.34, 95% CI 1.03-1.74, p = 0.028). Attention towards cardiac symptoms and symptom distress were positively associated with fear of PA (p < 0.001), which explained higher fear of PA in patients with a vigilant (directing attention towards health threats) coping style (p = 0.004).ConclusionsFear of PA assessed by the FActS-HF15 is a specific type of anxiety in patients with HF. Attention towards and being distressed by HF symptoms appear to play a central role in fear of PA, particularly in vigilant patients who are used to direct their attention towards health threats. These findings provide approaches for tailored interventions to reduce fear of PA and to increase PA in patients with HF.Trial registrationClinicalTrials.gov ID: NCT02898246.

Understanding how post-acute sequelae of SARS-CoV-2 infection (PASC) affects communities disproportionately affected by HIV is critically needed. This study aimed to identify the prevalence of PASC symptoms among Medicaid enrollees at risk for or living with HIV. Through a web survey, we received 138 valid responses from Medicaid-managed plan members who had received a COVID diagnosis. Participants' mean age was 45.4 years (SD = 11.9) and most were non-Hispanic Black (43.5%) or Hispanic (39.1%). Almost thirty-two percent reported inadequate incomes and 77.5% were HIV-positive. In the overall population, the frequently reported symptoms included neck/back/low back pain, brain fog/difficulty concentrating, bone/joint pain, muscle aches, and fatigue. Findings indicate that there is no statistically significant difference in the prevalence and intensity of PASC symptoms lasting 6 months or more between individuals living with and without HIV. Multiple regression analysis found that the number of PASC symptoms 6 months or longer was independently associated with inadequate incomes and comorbidities (cardiac problems, cancer, fibromyalgia) (R2 = .34). Those with inadequate incomes and comorbidities have more numerous PASC symptoms. Implications for health care delivery and long-term COVID services will be discussed.

Macadamia nuts constitute a vital component of both nutrition and livelihoods for smallholder producers in Malawi. We conducted a comprehensive mixed-methods study, combining qualitative and quantitative analyses, to explore varietal preferences and production challenges among these farmers. Leveraging cross-sectional data from 144 members of the Highlands Macadamia Cooperative Union Limited, our study underscores several significant findings. Our findings reveal that the majority of smallholder macadamia farmers (62%) are aged over 50, with farming as their primary occupation. Varied preferences are driven by yield-related traits, including high yield potential (38%), nut quality (29%), and extended flowering patterns (15%). Among the macadamia varieties, the top five choices, grown by over half of the farmers, include HAES 660 (18%), 800 (10%), 791 (9%), 816 (8%), and 246 (7%). Additionally, our study identifies five primary constraints faced by smallholder macadamia farmers: insect pests (81%), diseases (34%), limited market access (33%), wind damage (25%), and inadequate agricultural advisory services (17%). Based on these findings, we propose two policy recommendations to enhance smallholder macadamia production and productivity in Malawi and other regions. Specifically, we advocate for informed breeding programs that align with farmer preferences to promote greater adoption of macadamia varieties. Additionally, we emphasize the crucial role of the Malawian government in the macadamia value chain, suggesting active participation in providing extension services and marketing support, akin to its support for other cash crops.

Abstract The pathophysiology of degenerative cervical myelopathy (DCM) is characterized by chronic compression-induced damage to the spinal cord leading to secondary harm such as disruption of the blood spinal cord barrier (BSCB). It is therefore the purpose of this study to analyze BSCB disruption in pre- and postoperative DCM patients and to correlate those with the clinical status and postoperative outcome. This prospectively controlled cohort included 50 DCM patients (21 female; 29 male; mean age: 62.9 ± 11.2 years). As neurological healthy controls, 52 (17 female; 35 male; mean age 61.8 ± 17.3 years) patients with thoracic abdominal aortic aneurysm (TAAA) and indication for open surgery were included. All patients underwent a neurological examination and DCM-associated scores (Neck Disability Index, modified Japanese Orthopaedic Association Score) were assessed. To evaluate the BSCB status, blood and cerebrospinal fluid (CSF) samples (lumbar puncture or CSF drainage) were taken preoperatively and in 15 DCM patients postoperatively (4 female; 11 male; mean age: 64.7 ± 11.1 years). Regarding BSCB disruption, CSF and blood serum were examined for albumin, immunoglobulin (Ig) G, IgA and IgM. Quotients for CSF/serum were standardized and calculated according to Reiber diagnostic criteria. Significantly increased preoperative CSF/serum quotients were found in DCM patients as compared to control patients: AlbuminQ (p

Summary: Background: While vaccines have established utility against COVID-19, phase 3 efficacy studies have generally not comprehensively evaluated protection provided by previous infection or hybrid immunity (previous infection plus vaccination). Individual patient data from US government-supported harmonized vaccine trials provide an unprecedented sample population to address this issue. We characterized the protective efficacy of previous SARS-CoV-2 infection and hybrid immunity against COVID-19 early in the pandemic over three-to six-month follow-up and compared with vaccine-associated protection. Methods: In this post-hoc cross-protocol analysis of the Moderna, AstraZeneca, Janssen, and Novavax COVID-19 vaccine clinical trials, we allocated participants into four groups based on previous-infection status at enrolment and treatment: no previous infection/placebo; previous infection/placebo; no previous infection/vaccine; and previous infection/vaccine. The main outcome was RT-PCR-confirmed COVID-19 >7–15 days (per original protocols) after final study injection. We calculated crude and adjusted efficacy measures. Findings: Previous infection/placebo participants had a 92% decreased risk of future COVID-19 compared to no previous infection/placebo participants (overall hazard ratio [HR] ratio: 0.08; 95% CI: 0.05–0.13). Among single-dose Janssen participants, hybrid immunity conferred greater protection than vaccine alone (HR: 0.03; 95% CI: 0.01–0.10). Too few infections were observed to draw statistical inferences comparing hybrid immunity to vaccine alone for other trials. Vaccination, previous infection, and hybrid immunity all provided near-complete protection against severe disease. Interpretation: Previous infection, any hybrid immunity, and two-dose vaccination all provided substantial protection against symptomatic and severe COVID-19 through the early Delta period. Thus, as a surrogate for natural infection, vaccination remains the safest approach to protection. Funding: National Institutes of Health.

Abstract Iron deficiency (ID) is the most common nutritional disorder worldwide. It is often observed in patients with chronic diseases, such as heart failure (HF), chronic kidney disease (CKD), inflammatory bowel disease (IBD) and cancer. ID is associated with poor clinical outcome, including poor performance, reduced quality of life, as well as increased hospitalization and mortality. The aim of this review is to provide an overview about the role of ID in chronic diseases (HF, CKD, IBD, cancer) regarding their current definitions and clinical relevance; diagnostic accuracy of iron parameters in chronic inflammatory conditions and its potential as prognostic markers. Due to different definitions and guideline recommendations of ID, various laboratory parameters for ID diagnostic exist and there is no general consensus about the definition of ID and its treatment. Still, a general trend can be observed across all investigated indications of this review (HF, CKD, IBD, cancer) that serum ferritin and transferrin saturation (TSAT) are the two parameters mentioned most often and emphasized in all guidelines to define ID and guide treatment. The most commonly used threshold values for the diagnosis of ID are TSAT of

Abstract Fused filament fabrication (FFF) represents a straightforward additive manufacturing technique applied in the medical sector for personalized patient treatment. However, frequently processed biopolymers lack sufficient thermal stability to be used as auxiliary devices such as surgical guides. The aim of this study was to evaluate the dimensional accuracy of experimental biocopolyester blends with improved thermal characteristics after printing, annealing and sterilization. A total of 160 square specimens and 40 surgical guides for oral implant placement were printed. One subgroup of each material (n = 10) underwent thermal annealing before both subgroups were subjected to steam sterilization (134 °C; 5 min). Specimens were digitized and the deviation from the original file was calculated. The thermal behavior was analyzed using differential scanning calorimetry and thermogravimetric analysis. A one-way ANOVA and t-tests were applied for statistical analyses (p

Abstract Smoking is a leading risk factor for many of the top ten causes of death worldwide. Of the 1.3 billion smokers globally, 80% live in low- and middle-income countries, where the number of deaths due to tobacco use is expected to double in the next decade according to the World Health Organization. Genetic studies have helped to identify biological pathways for smoking behaviours, but have mostly focussed on individuals of European ancestry or living in either North America or Europe. We performed a genome-wide association study of two smoking behaviour traits in 10,558 men of African ancestry living in five African countries and the UK. Eight independent variants were associated with either smoking initiation or cessation at P-value

Abstract Monoclonal antibodies (abs) targeting the programmed cell death 1 (PD-1) immune checkpoint pathway have revolutionized tumor therapy. Because T-cell-directed PD-1 blockade boosts tumor immunity, anti-PD-1 abs have been developed for examining T-cell-PD-1 functions. More recently, PD-1 expression has also been reported directly on cancer cells of various etiology, including in melanoma. Nevertheless, there is a paucity of studies validating anti-PD-1 ab clone utility in specific assay types for characterizing tumor cell-intrinsic PD-1. Here, we demonstrate reactivity of several anti-murine PD-1 ab clones and recombinant PD-L1 with live B16-F10 melanoma cells and YUMM lines using multiple independent methodologies, positive and negative PD-1-specific controls, including PD-1-overexpressing and PD-1 knockout cells. Flow cytometric analyses with two separate anti-PD-1 ab clones, 29F.1A12 and RMP1-30, revealed PD-1 surface protein expression on live murine melanoma cells, which was corroborated by marked enrichment in PD-1 gene (Pdcd1) expression. Immunoblotting, immunoprecipitation, and mass spectrometric sequencing confirmed PD-1 protein expression by B16-F10 cells. Recombinant PD-L1 also recognized melanoma cell-expressed PD-1, the blockade of which by 29F.1A12 fully abrogated PD-1:PD-L1 binding. Together, our data provides multiple lines of evidence establishing PD-1 expression by live murine melanoma cells and validates ab clones and assay systems for tumor cell-directed PD-1 pathway investigations.

Abstract Calpains are calcium-activated neutral proteases involved in the regulation of key signaling pathways. Junctophilin-2 (JP2) is a Calpain-specific proteolytic target and essential structural protein inside Ca2+ release units required for excitation-contraction coupling in cardiomyocytes. While downregulation of JP2 by Calpain cleavage in heart failure has been reported, the precise molecular identity of the Calpain cleavage sites and the (patho-)physiological roles of the JP2 proteolytic products remain controversial. We systematically analyzed the JP2 cleavage fragments as function of Calpain-1 versus Calpain-2 proteolytic activities, revealing that both Calpain isoforms preferentially cleave mouse JP2 at R565, but subsequently at three additional secondary Calpain cleavage sites. Moreover, we identified the Calpain-specific primary cleavage products for the first time in human iPSC-derived cardiomyocytes. Knockout of RyR2 in hiPSC-cardiomyocytes destabilized JP2 resulting in an increase of the Calpain-specific cleavage fragments. The primary N-terminal cleavage product NT1 accumulated in the nucleus of mouse and human cardiomyocytes in a Ca2+-dependent manner, closely associated with euchromatic chromosomal regions, where NT1 is proposed to function as a cardio-protective transcriptional regulator in heart failure. Taken together, our data suggest that stabilizing NT1 by preventing secondary cleavage events by Calpain and other proteases could be an important therapeutic target for future studies.

The purpose of this study was to analyze the Sources of Strength program implemented at a high school in the southeastern United States using the RE-AIM framework. Students (n = 1,250) were recruited from one high school in the southeastern United States that was implementing Sources of Strength. College undergraduate students mentored high school peer leaders to implement Sources of Strength to promote opioid and suicide protective factors for the high school. This was a cross-sectional design in which a post-implementation survey assessed program effectiveness, satisfaction, and feedback for maintenance. Analysis used the RE-AIM framework of reach, effectiveness, adoption, implementation, and maintenance. Reach: 100 % of students participated in at least one campaign. Effectiveness: Mean protective factor scores were 3.83 for happiness; 3.93 for knowledge of coping skills, and 3.38 for practice of coping skills. Adoption: One high school and 100 % of classrooms participated in at least one campaign. Implementation: 18 high school peer leader meetings were facilitated by the undergraduate mentors. Maintenance: High school students qualitatively provided input on future campaign needs, and the program was continued following this RE-AIM analysis. The Sources of Strength program promoted suicide and opioid protective factors in high school students during COVID-19. Future research should compare the efficacy of Sources of Strength at baseline and follow-up with multiple high school sites.

Abstract Background Allied Health Professionals (AHPs) commonly use journal clubs (JCs) to support Evidence-Based Practice (EBP). There is however little research regarding implementing and sustaining JCs in the long term, and their impact on EBP use and skills in AHPs. This study investigated the impact of implementing a structured JC format, called “TREAT” (previously only investigated across 6 sessions), over a longer period of 16 sessions for AHPs in a public health service. The study also investigated AHP’s attendance, adherence, satisfaction and barriers and enablers to implementing the format. Methods A mixed methods hybrid-effectiveness implementation design was employed, guided by the Knowledge-to-Action cycle. EBP skills, confidence, use, and attitudes were assessed (Adapted Fresno Test, EBPQ, tailored journal club culture questionnaire) at baseline, and after 10 and 16-monthly sessions. Satisfaction and impact on clinical practice were explored using questionnaires at 10 and 16-months, with free-form responses identifying enablers and barriers to EBP culture and implementation. Data on attendance and adherence to the TREAT format were also collected. Results Six JCs comprising a total of 132 unique participants from seven Allied Health professions were assessed across three time points. EBP skills improved on the Adapted Fresno Test after 10-monthly (6.6 points: 95% CI, 0.43 to 12.7) and 16-monthly sessions (7.8 points, 95% CI, 0.85 to 14.7), and on self-reported total EBPQ ratings of confidence at 10-months (4.9 points: 95% CI, 2.2 to 7.5) and 16-months (5.7 points: 95% CI 2.7 to 8.7). Of 132 AHPs, 88 reported adopting new treatments/resources and 64 reported updating clinical procedures. Mean attendance was 5.7 sessions (SD = 3.8). Adherence to TREAT components in each session was 86% (95% CI, 83% to 89%). Most participants recommended the format and reported a desire to continue. Enablers to the JC included using clinically relevant topics and active participation while reported barriers included limited time to prepare. Conclusions Despite variable attendance, TREAT JCs can continue to be implemented within a service for 16 monthly-sessions, and may contribute to improved EBP skills and confidence and changes in clinical practice over time. Tailoring of implementation strategies was shown to be important to address local enablers and barriers.

Abstract For image-guided small animal irradiations, the whole workflow of imaging, organ contouring, irradiation planning, and delivery is typically performed in a single session requiring continuous administration of anaesthetic agents. Automating contouring leads to a faster workflow, which limits exposure to anaesthesia and thereby, reducing its impact on experimental results and on animal wellbeing. Here, we trained the 2D and 3D U-Net architectures of no-new-Net (nnU-Net) for autocontouring of the thorax in mouse micro-CT images. We trained the models only on native CTs and evaluated their performance using an independent testing dataset (i.e., native CTs not included in the training and validation). Unlike previous studies, we also tested the model performance on an external dataset (i.e., contrast-enhanced CTs) to see how well they predict on CTs completely different from what they were trained on. We also assessed the interobserver variability using the generalized conformity index ( $$\hbox {CI}_{\mathrm{gen}}$$ CI gen ) among three observers, providing a stronger human baseline for evaluating automated contours than previous studies. Lastly, we showed the benefit on the contouring time compared to manual contouring. The results show that 3D models of nnU-Net achieve superior segmentation accuracy and are more robust to unseen data than 2D models. For all target organs, the mean surface distance (MSD) and the Hausdorff distance (95p HD) of the best performing model for this task (nnU-Net 3d_fullres) are within 0.16 mm and 0.60 mm, respectively. These values are below the minimum required contouring accuracy of 1 mm for small animal irradiations, and improve significantly upon state-of-the-art 2D U-Net-based AIMOS method. Moreover, the conformity indices of the 3d_fullres model also compare favourably to the interobserver variability for all target organs, whereas the 2D models perform poorly in this regard. Importantly, the 3d_fullres model offers 98% reduction in contouring time.

Abstract Elevated fibroblast growth factor 23 (FGF23) levels are associated with adverse outcome in populations with cardiovascular disease and chronic kidney failure. It is unclear if FGF23 has significance in prognosis estimation in patients with acute heart failure (HF) when compared to traditional risk estimation tools. Serum levels of intact FGF23 were assessed in 139 patients admitted to the Intermediate Care Unit of a tertiary hospital for acute HF. Patients were followed-up for one year. After exclusion of patients who were lost to follow-up, data outliers, and patients with sampling errors, the final study cohort comprised 133 patients. The Seattle Heart Failure (SHF) Model was used to estimate one-year survival. FGF23 levels correlated with HF severity and were strongly associated with one-year mortality. Associations between one-year outcome and FGF23, assessed on day 1 after admission, were still evident after multivariable adjustment (OR 15.07; 95%CI 1.75–129.79; p = 0.014). FGF23 levels predicted the one-year outcome with similar accuracy as the SHF Model, both if assessed on day 1 and on day 2 after admission (FGF23d1: AUC 0.784; 95%CI 0.669–0.899; FGF23d2: AUC 0.766; 95%CI 0.631–0.901; SHF: AUC 0.771; 95%CI 0.651–0.891). The assessment of FGF23 in patients with acute HF might help identify high-risk patients that are more prone to complications, need a closer follow-up and more aggressive treatment.

Abstract Cellular responses to DNA double-strand breaks (DSBs) not only promote genomic integrity in healthy tissues, but also largely determine the efficacy of many DNA-damaging cancer treatments, including X-ray and particle therapies. A growing body of evidence suggests that activation of the mechanisms that detect, signal and repair DSBs may depend on the complexity of the initiating DNA lesions. Studies focusing on this, as well as on many other radiobiological questions, require reliable methods to induce DSBs of varying complexity, and to visualize the ensuing cellular responses. Accelerated particles of different energies and masses are exceptionally well suited for this task, due to the nature of their physical interactions with the intracellular environment, but visualizing cellular responses to particle-induced damage - especially in their early stages - at particle accelerator facilities, remains challenging. Here we describe a straightforward approach for real-time imaging of early response to particle-induced DNA damage. We rely on a transportable setup with an inverted fluorescence confocal microscope, tilted at a small angle relative to the particle beam, such that cells can be irradiated and imaged without any microscope or beamline modifications. Using this setup, we image and analyze the accumulation of fluorescently-tagged MDC1, RNF168 and 53BP1—key factors involved in DSB signalling—at DNA lesions induced by 254 MeV α-particles. Our results provide a demonstration of technical feasibility and reveal asynchronous initiation of accumulation of these proteins at different individual DSBs.

Abstract By using gold (Au) nanoparticles (NPs) as an optical near-field source under far-field illumination in combination with a silver (Ag) ion solution containing a photoinitiator, we coated Ag on Au NPs using a near-field (NF)-assisted process. We evaluated the change in the size of the NPs using transmission electron microscopy. Evaluation of the synthesized Ag volume over illumination power confirmed the squared power dependence of the NP volume with illumination using 808 nm light, i.e., a wavelength longer than the absorption edge wavelength of the photoinitiator molecules. The rate of volume increase was much lower than the plasmonic field enhancement effect. Therefore, the squared power dependency of the volume increase using a wavelength longer than the absorption edge wavelength originated from NF-assisted second-harmonic generation and the resulting excitation.

Abstract Endogenous immune mediated reactions of inflammation and angiogenesis are components of the spinal cord injury in patients with degenerative cervical myelopathy (DCM). The aim of this study was to identify alteration of certain mediators participating in angiogenetic and inflammatory reactions in patients with DCM. A consecutive series of 42 patients with DCM and indication for surgical decompression were enrolled for the study. 28 DCM patients were included, as CSF samples were taken preoperatively. We enrolled 42 patients requiring surgery for a thoracic abdominal aortic aneurysm (TAAA) as neurologically healthy controls. In 38 TAAA patients, CSF samples were taken prior to surgery and thus included. We evaluated the neurological status of patients and controls prior to surgery including NDI and mJOA. Protein-concentrations of factors with a crucial role in inflammation and angiogenesis were measured in CSF via ELISA testing (pg/ml): Angiopoietin 2, VEGF-A and C, RANTES, IL 1 beta and IL 8. Additionally, evaluated the status of the blood-spinal cord barrier (BSCB) by Reibers´diagnostic in all participants. Groups evidently differed in their neurological status (mJOA: DCM 10.1 ± 3.3, TAAA 17.3 ± 1.2, p

Abstract Background In patients with iron deficiency anemia, ferric carboxymaltose (FCM) and ferric derisomaltose (FDI) allow high-dose iron repletion. While FCM is reported to induce hypophosphatemia, the frequency of hypophosphatemia after an equivalent dosage of FDI had not been assessed prospectively. Methods In the prospective, single-center, double-blind HOMe aFers study, 26 women with iron deficiency anemia (hemoglobin

BackgroundPulmonary Tuberculosis (TB) is diagnosed through sputum samples. As sputum sampling is challenging in children and cachexic patients, the development of diagnostic tests using saliva appears promising but has been discouraged due to low bacterial load and poor sensitivity. Here, we present a novel and rapid method to enrich Mycobacterium tuberculosis (Mtb) from saliva, which may serve as a basis for a diagnostic saliva test.MethodsLipobiotin-functionalized magnetic beads (LMBs) were incubated with Mtb-spiked PBS and saliva from healthy donors as well as with saliva from TB patients. Flow cytometry was used to evaluate the capacity of the beads to bind Mtb, while real-time quantitative polymerase chain reaction (qPCR) was utilized to detect Mtb and determine the amount of mycobacterial DNA in different sample types.ResultsWe found that LMBs bind Mtb efficiently when compared to non-functionalized beads. The development of an qPCR assay based on the use of LMBs (LMB assay) allowed us to enrich mycobacterial DNA in spiked sample types, including PBS and saliva from healthy donors (enrichment of up to ~8.7 fold). In Mtb-spiked saliva samples, we found that the LMB assay improved the detection rate of 102 bacteria in a volume of 5 ml from 0 out of 15 (0%) to 6 out of 15 (40%). Consistent with that, the LMB assay increased the rate of correctly identified saliva samples from TB patients in two independent cohorts.ConclusionsImplementation of the principle of the LMB-based assay may improve the sensitivity of existing diagnostic techniques, e.g. by functionalizing materials that facilitate Mtb sampling from the oral cavity.

Tick-borne encephalitis (TBE) is Eurasia’s most important tick-borne viral disease. Rodents play an important role as natural hosts. Longitudinal studies on the dynamics of the seroprevalence rates in wild rodents in natural foci over the year are rare, and the dynamics of the transmission cycle still need to be understood. To better understand the infection dynamics, rodents were captured in a capture-mark-release-recapture-study in two natural foci in Bavaria, Germany, monthly from March 2019 to October 2022. Overall, 651 blood and thoracic lavage samples from 478 different wild rodents (Clethrionomys glareolus and Apodemus flavicollis) were analyzed for antibodies against tick-borne encephalitis virus (TBEV) by indirect immunofluorescence assay (IIFA) and confirmed using a serum neutralization test (SNT). Furthermore, a generalized linear mixed model (GLMM) analysis was performed to investigate ecological and individual factors for the probability of infection in rodents. Clethrionomys glareolus (19.4%) had a higher seroprevalence than A. flavicollis (10.5%). Within Cl. glareolus, more males (40.4%) than females (15.6%) were affected, and more adults (25.4%) than juveniles (9.8%). The probability of infection of rodents rather depends on factors such as species, sex, and age than on the study site of a natural focus, year, and season. The high incidence rates of rodents, particularly male adult bank voles, highlight their critical role in the transmission cycle of TBEV in a natural focus and demonstrate that serologically positive rodents can be reliably detected in a natural focus regardless of season or year. In addition, these data contribute to a better understanding of the TBEV cycle and thus could improve preventive strategies for human infections.

Background Antigen-based point-of-care tests for identification of SARS-CoV-2 may markedly enhance effectiveness of population-based controlling strategies. Previous studies have demonstrated >70% sensitivity and high specificity compared with reverse transcriptase real-time PCR (RT-PCR) in symptomatic individuals, but test performance for asymptomatic individuals is unknown.Methods Test performance of the Panbio COVID-19 Ag Rapid Test (Abbott) was compared with RT-PCR in a longitudinal cohort study of asymptomatic football players and staff members of professional football clubs. Based on timing of symptoms and prior and subsequent test results, positive RT-PCR tests were categorised as presymptomatic, early or late infection, or persistent RNA shedding.Findings 2425 tests were performed in 824 individuals, of which 52 (6.3%) were SARS-CoV-2 positive based on RT-PCR. There were 2406 paired sets from asymptomatic subjects for analysis. Sixteen Panbio tests were inconclusive, for which sensitivity analyses were performed (considering results as either positive or negative or being excluded). Sensitivity of Panbio for screening of asymptomatic individuals ranged from 80.0% (61.4–92.3) to 86.67% (69.2–96.2) and specificity from 99.53% (95% CI 99.2 to 99.8) to 100% (95% CI 99.8 to 100). Sensitivity of Panbio to detect subjects with presymptomatic/early infection (n=42) ranged from 81.82% (95% CI 67.3 to 91.8) to 90.91% (95% CI 78.3 to 97.5) with specificity always above 99%.Interpretation The Panbio COVID-19 Ag rapid test identifies 81%–90% of presymptomatic and early asymptomatic SARS-CoV-2 infections with high specificity. This test may therefore be adopted in testing strategies such as targeted screening of specific populations where prevalence is low.

Genome-wide association studies (GWAS) provide huge information on statistically significant single-nucleotide polymorphisms (SNPs) associated with various human complex traits and diseases. By performing GWAS studies, scientists have successfully identified the association of hundreds of thousands to millions of SNPs to a single phenotype. Moreover, the association of some SNPs with rare diseases has been intensively tested. However, classic GWAS studies have not yet provided solid, knowledgeable insight into functional and biological mechanisms underlying phenotypes or mechanisms of diseases. Therefore, several post-GWAS (pGWAS) methods have been recommended. Currently, there is no simple scientific document to provide a quick guide for performing pGWAS analysis. pGWAS is a crucial step for a better understanding of the biological machinery beyond the SNPs. Here, we provide an overview to performing pGWAS analysis and demonstrate the challenges behind each method. Furthermore, we direct readers to key articles for each pGWAS method and present the overall issues in pGWAS analysis. Finally, we include a custom pGWAS pipeline to guide new users when performing their research.

OBJECTIVES This study evaluated the effectiveness of hydrogen peroxide (H2O2) as mouthwash and nasal spray on symptom relief in coronavirus disease 2019 (COVID-19) patients. METHODS Patients positive for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), who were treated in a hospital or at home, and patients’ family members (not positive for SARS-CoV-2), were randomized into 2 groups: experimental (1% H2O2 for gargling, 0.5% H2O2 for nasal wash), and control. Patients gargled the solution 3 times a day, and applied the nasal spray twice a day, for a 7-day period. Family members received the same treatment as the treated COVID-19 patient. The researchers contacted patients every 2 days over an 8-day period. An average post-treatment interval of 8 days passed before testing family members. RESULTS The most frequent symptoms on day 0 were cough, loss of taste, and hyposmia; there were no significant differences between groups, independent of the period. The symptom of dyspnea presented a significant difference between days 2 and 4 (p

OBJECTIVES To evaluate the effectiveness of hydrogen peroxide (H2O2) in the form of mouthwash and nasal spray as an auxiliary treatment for coronavirus disease 2019 (COVID-19). METHODS Forty hospitalized patients who tested positive for severe acute respiratory syndrome coronavirus 2 using a reverse-transcription polymerase chain reaction test were evaluated. They were randomly divided into an experimental group (n= 20; gargling with 1.0% H2O2 and nasal wash with 0.5% H2O2) or a control group (n= 20). The solutions were used for 7 days and the patients were monitored every 2 days, for a total of 8 days. At check-ups, patients were asked about their symptoms and possible adverse effects of the solutions. The presence and severity (mild, moderate, or severe) of symptoms were recorded. Data were compared using the Student test and the Fisher exact test (α= 0.05). RESULTS There was no significant difference between the 2 groups in the length of hospital stay (p= 0.65). The most frequent symptom on day 0 was coughing (72.0% in the experimental group and 76.5% in the control group), which abated over time. There was no significant difference between the groups in the evaluated symptoms. Most (75.0%) of the patients in the experimental group presented a reduction in dyspnea between days 0 and 2. Few patients reported adverse effects from the use of the solutions. CONCLUSIONS H2O2 as a mouthwash and nasal spray is safe to use. There is insufficient evidence to demonstrate that H2O2 is effective as an auxiliary treatment for hospitalized COVID-19 patients.

ObjectivesPatients do better in research-intense environments. The importance of research is reflected in the accreditation requirements of Australian clinical specialist colleges. The nature of college-mandated research training has not been systematically explored. We examined the intended research curricula of Australian trainee doctors described by specialist colleges, their constructive alignment and the nature of scholarly project requirements.DesignWe undertook content analysis of publicly available documents to characterise college research training curricula.SettingWe reviewed all publicly accessible information from the websites of Australian specialist colleges and their subspecialty divisions. We retrieved curricula, handbooks and assessment-related documents.ParticipantsFifty-eight Australian specialist colleges and their subspecialty divisions.Primary and secondary outcome measuresTwo reviewers extracted and coded research-related activities as learning outcomes, activities or assessments, by research stage (using, participating in or leading research) and competency based on Bloom’s taxonomy (remembering, understanding, applying, analysing, evaluating, creating). We coded learning and assessment activities by type (eg, formal research training, publication) and whether it was linked to a scholarly project. Requirements related to project supervisors’ research experience were noted.ResultsFifty-five of 58 Australian college subspecialty divisions had a scholarly project requirement. Only 11 required formal research training; two required an experienced research supervisor. Colleges emphasised a role for trainees in leading research in their learning outcomes and assessments, but not learning activities. Less emphasis was placed on using research, and almost no emphasis on participation. Most learning activities and assessments mapped to the ‘creating’ domain of Bloom’s taxonomy, whereas most learning outcomes mapped to the ‘evaluating’ domain. Overall, most research learning and assessment activities were related to leading a scholarly project.ConclusionsAustralian specialist college research curricula appear to emphasise a role for trainees in leading research and producing research deliverables, but do not mandate formal research training and supervision by experienced researchers.

BACKGROUND:Aortic stenosis (AS) may lead to diastolic dysfunction and later on heart failure (HF) with preserved left ventricular ejection fraction (HFpEF) via increased afterload and left-ventricular (LV) hypertrophy. Since epicardial adipose tissue (EAT) is a metabolically active fat depot that is adjacent to the myocardium and can influence cardiomyocytes and LV function via secretion of proinflammatory cytokines, we hypothesized that high amounts of EAT, as assessed by computed tomography (CT), may aggravate the development and severity of LV hypertrophy and diastolic dysfunction in the context of AS. METHODS:We studied 50 patients (mean age 71 ± 9 years; 9 women) in this preliminary study with mild or moderate AS and mild to severe LV diastolic dysfunction (LVDD), diagnosed by echocardiography, who underwent non-contrast cardiac CT and echocardiography. EAT parameters were measured on 2nd generation dual source CT. Conventional two-dimensional echocardiography and Tissue Doppler Imaging (TDI) was performed to assess LV function and to derive myocardial straining parameter. All patients had a preserved LV ejection fraction > 50%. Data was analysed using Pearson's correlation. RESULTS:Only weak correlation was found between EAT volume or density and E/é ratio as LVDD marker (r = -.113 p = .433 and r = .260, p = .068 respectively). Also, EAT volume or density were independent from Global Strain Parameters (r = 0.058 p = .688 and r = -0.207 p = .239). E/é ratio was strongly associated with LVDD (r = .761 p≤0.0001) and Strain Parameters were moderately associated with LV Ejection Fraction (r = -.669 p≤0.001 and r = -.454 P≤0.005). CONCLUSIONS:In this preliminary study in patients with AS, the EAT volume and density as assessed by CT correlated only weakly with LVDD, as expressed by the commonly used E/é ratio, and with LV strain function. Hence, measuring EAT volume and density may neither contribute to the prediction nor upon the severity of LVDD, respectively.

Abstract The nitrogen-vacancy (NV) centre in diamond is a promising candidate for quantum computing applications and magnetic sensing applications, because it is an atomic-scale defect with stable coherence time (T 2) and reliable accessibility at room temperature. We demonstrated a method for improving the NV spin properties (the full width half maximum (FWHM) value of the magnetic resonance spectrum and T 2) through a near-field (NF) etching method under ambient conditions. The NF etching method, based on a He-Cd ultraviolet laser (325 nm), which is longer than the absorption edge of the oxygen molecule, enabled selective removal of defects on the nanodiamond surface. We observed a decrease in the FWHM value close to 15% and an increase in T 2 close to 25%. Since our technique can be easily reproduced, a wide range of NV centre applications could be improved, especially magnetic sensing applications. Our results are especially attractive, because they have been obtained under ambient conditions and only require a light source with wavelength slightly above the O2 absorption edge.

Se relata el caso de un paciente masculino de 56 años con mieloma múltiple (MM) y presencia de plasmocitomas en tejido celular subcutáneo y en testículo. El paciente tenía el antecedente de MM tratado con quimioterapia y trasplante autólogo de médula ósea y que después de 4 años de remisión de la enfermedad presentó recidiva ósea, la misma que fue tratada con quimioterapia e inmunoterapia. No hubo respuesta satisfactoria a los primeros ciclos del tratamiento con progresión de la enfermedad ósea y surgimiento de nódulos en dorso, puño y testículo sospechosos de recidiva de la enfermedad. Se procedió a resección de las lesiones en el dorso y puño que, en el examen anatomopatológico, fueron compatibles con neoplasia maligna y la prueba de inmunohistoquímica confirmó que se trataba de metástasis del MM. Se optó por la continuidad del tratamiento hasta completar los 12 ciclos propuestos, pero no hubo mejoría de las lesiones además de la aparición de nuevas lesiones en tejido celular subcutáneo y región perianal, con empeoramiento clínico. El paciente falleció después de dos meses de cuidados paliativos.

Abstract Background Antimicrobial peptides are important components of the host defence with a broad range of functions including direct antimicrobial activity and modulation of inflammation. Lack of cathelin-related antimicrobial peptide (CRAMP) was associated with higher mortality and bacterial burden and impaired neutrophil granulocyte infiltration in a model of pneumococcal meningitis. The present study was designed to characterize the effects of CRAMP deficiency on glial response and phagocytosis after exposure to bacterial stimuli. Methods CRAMP-knock out and wildtype glial cells were exposed to bacterial supernatants from Streptococcus pneumoniae and Neisseria meningitides or the bacterial cell wall components lipopolysaccharide and peptidoglycan. Cell viability, expression of pro- and anti-inflammatory mediators and activation of signal transduction pathways, phagocytosis rate and glial cell phenotype were investigated by means of cell viability assays, immunohistochemistry, real-time RT-PCR and Western blot. Results CRAMP-deficiency was associated with stronger expression of pro-inflammatory and weakened expression of anti-inflammatory cytokines indicating a higher degree of glial cell activation even under resting-state conditions. Furthermore, increased translocation of nuclear factor ‘kappa-light-chain-enhancer’ of activated B-cells was observed and phagocytosis of S. pneumoniae was reduced in CRAMP-deficient microglia indicating impaired antimicrobial activity. Conclusions In conclusion, the present study detected severe alterations of the glial immune response due to lack of CRAMP. The results indicate the importance of CRAMP to maintain and regulate the delicate balance between beneficial and harmful immune response in the brain.

The stalk domain of the hemagglutinin has been identified as a target for induction of protective antibody responses due to its high degree of conservation among numerous influenza subtypes and strains. However, current assays to measure stalk-based immunity are not standardized. Hence, harmonization of assay readouts would help to compare experiments conducted in different laboratories and increase confidence in results. Here, serum samples from healthy individuals (n = 110) were screened using a chimeric cH6/1 hemagglutinin enzyme-linked immunosorbent assay (ELISA) that measures stalk-reactive antibodies. We identified samples with moderate to high IgG anti-stalk antibody levels. Likewise, screening of the samples using the mini-hemagglutinin (HA) headless construct #4900 and analysis of the correlation between the two assays confirmed the presence and specificity of anti-stalk antibodies. Additionally, samples were characterized by a cH6/1N5 virus-based neutralization assay, an antibody-dependent cell-mediated cytotoxicity (ADCC) assay, and competition ELISAs, using the stalk-reactive monoclonal antibodies KB2 (mouse) and CR9114 (human). A “pooled serum” (PS) consisting of a mixture of selected serum samples was generated. The PS exhibited high levels of stalk-reactive antibodies, had a cH6/1N5-based neutralization titer of 320, and contained high levels of stalk-specific antibodies with ADCC activity. The PS, along with blinded samples of varying anti-stalk antibody titers, was distributed to multiple collaborators worldwide in a pilot collaborative study. The samples were subjected to different assays available in the different laboratories, to measure either binding or functional properties of the stalk-reactive antibodies contained in the serum. Results from binding and neutralization assays were analyzed to determine whether use of the PS as a standard could lead to better agreement between laboratories. The work presented here points the way towards the development of a serum standard for antibodies to the HA stalk domain of phylogenetic group 1.

Eutrophication has played a major role in the worldwide increase of harmful algal blooms (HABs). Higher input of key nutrients, such as nitrogen (N) and phosphorus (P), can stimulate the growth of harmful algal species in freshwater, estuarine, and coastal marine ecosystems. Some HAB-forming taxa, particularly several cyanobacteria and dinoflagellate species, are harmful through the production of N-rich toxins that have detrimental effects on the environment and human health. Here, we test how changes in nutrient availability affect N-rich toxin synthesis in cyanobacteria and dinoflagellates using a meta-analysis approach. Overall, N-rich toxin content showed an increase with P limitation, while it tended to decrease with N limitation, but we also observed substantial variation in responses both within and across genera and toxin groups. For instance, in response to N limitation, microcystin content varied from a 297% decrease up to a 273% increase, and paralytic shellfish poisoning (PSP) toxin content varied from a 204% decrease to an 82% increase. Cylindrospermopsin, produced by N2-fixing cyanobacteria, showed no clear direction in response to nutrient limitation, and cellular contents of this compound may thus vary independently of nutrient fluctuations. Our results confirm earlier reported stoichiometric regulation of N-rich phytoplankton toxins, showing increased toxin content with an increase in cellular N:P ratios, and vice versa. Thus, changes in N-rich toxin content largely follow the changes in relative cellular N content. Consequently, although nutrient limitation may limit bloom biomass and thereby bloom toxicity, our results warn that P limitation can cause accumulation of cellular toxins and thus lead to unexpected increases in bloom toxicity.