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Abstract Background Harmonizing core outcome domains allows for pooling data, comparing interventions, and streamlining research evaluation. At the same time clinicians require concise and feasible measures for routine practice. Considering the heterogeneity of post-COVID-19 condition, a biopsychosocial approach requires sufficient coverage of the psychosocial dimension with assessments. Previous recommendations for core outcome sets have serious limitations regarding the psychosocial aspects of post-COVID-19 condition. This paper specifically focuses on psychosocial outcomes for adults with post-COVID-19 condition, providing both a comprehensive set of outcome domains for research and a streamlined clinical core set tailored for routine clinical use. Methods In a structured Consensus Development Approach, the European Network to improve diagnostic, treatment, and healthcare for patients with persistent somatic symptoms (EURONET-SOMA) developed psychosocial core outcome domains and assessments regarding post-COVID-19 condition. The experts identified variables and instruments which should be considered in studies on adults suffering from post-COVID-19 condition, and which are feasible in the clinical setting and relevant for research. Results We identified three higher-order dimensions with each encompassing several domains: The first higher-order dimension, “outcomes”, encompasses (1) the classification/ diagnostics of post-COVID-19 condition, (2) somatic symptoms (including fatigue), (3) the psychopathological status and mental comorbidities, (4) the physical status and somatic comorbidities, (5) neurocognitive symptoms, and (6) illness consequences. The second higher-order domain “mechanisms” encompasses (7) cognitive components, (8) affective components, (9) behavioral components, (10) social components, and (11) psychobiological bridge markers (e.g., neuroimmunological and psychoneuroendocrinological variables). The third higher-order domain, “risk factors”, includes factors such as (12) socioeconomic status and sociocultural factors, (13) pre-existing mental and somatic health issues, (14) personality factors (e.g., neuroticism), (15) adverse childhood experiences, (16) ongoing disability or pension claim, and (17) social media use. For each domain, specific instruments are suggested for research purposes and clinical use. Conclusions The recommended core domains help to increase consistency in a biopsychosocial approach to post-COVID-19 condition across investigations, improve synergies, and facilitate decision-making when comparing different interventional approaches. It allows to better identify relevant subgroups in heterogeneous post-COVID-19 condition populations offering practical tools for routine clinical practice through the clinical core set.

Abstract Successful implementation of remote monitoring of vital signs outside of the hospital setting hinges on addressing three crucial unmet needs: longer-term wear, skin comfort and signal quality. Earlier, we developed a Health Patch research platform that uses self-adhesive dry electrodes to measure vital digital biomarkers. Here, we report on the analytical validation for heart rate, heart rate variability and respiration rate. Study design included n = 25 adult participants with data acquisition during a 30-minute exercise protocol involving rest, squats, slow, and fast cycling. The Shimmer3 ECG Unit and Cosmed K5, were reference devices. Data analysis showed good agreement in heart rate and marginal agreement in respiratory rate, with lower agreement towards higher respiratory rates. The Lin’s concordance coefficient was 0.98 for heart rate and 0.56 for respiratory rate. Heart rate variability (RMSSD) had a coefficient of 0.85. Participants generally expressed a positive experience with the technology, with some minor irritation from the medical adhesive. The results highlighted potential of this technology for short-to-medium term clinical use for cardiorespiratory health, due to its reliability, accuracy, and compact design. Such technology could become instrumental for remote monitoring providing healthcare professionals with continuous data, remote assessment and enhancing patient outcomes in cardiorespiratory health management.

Abstract Attachment theory is one of the most comprehensive frameworks in social and developmental psychology. It describes how selective, enduring emotional bonds between infants and their caregivers are formed and maintained throughout life. These attachment bonds exhibit distinct characteristics that are intimately tied to fundamental aspects of mammalian life, including pregnancy, birth, lactation, and infant brain development. However, there is a lack of a cohesive biological narrative that explains the psychological forces shaping attachment behavior and the emergence and consolidation of attachment patterns at a neurobiological level. Here, we propose a theoretical narrative focusing on organized attachment patterns that systematically link the two primary purposes of the attachment behavioral system: the provision of tangible protection or support and the corresponding subjective feeling of safety or security. We aim for this detailed delineation of neurobiological circuits to foster more comprehensive and interdisciplinary future research.

ABSTRACT This study evaluated initial antihypertensive drug prescription patterns in Indian healthcare settings. An observational, cross‐sectional, prospective prescription registry analyzed prescriptions for 4723 newly diagnosed hypertension patients. Additionally, it investigated the extent to which physicians adhered to either European or Indian hypertension guidelines. Angiotensin receptor blockers (ARBs) were the most commonly prescribed drugs, given to 79% of patients, followed by calcium channel blockers (CCBs) at 55%. Diuretics and beta‐blockers (BBs) were prescribed to 27% and 17% of patients, respectively. Monotherapy was administered to 35% of patients, while combination therapies were more prevalent, with dual therapy at 51% and regimens involving three or more drugs prescribed to 14%. Among multi‐drug treatments (n = 3082, 65%), 98% received fixed‐dose combination tablets. The most common combinations were ARB + CCB (26%), ARB + diuretic (12%), and ARB + CCB + diuretic (8%). Key predictors for an increasing number of prescribed drugs included statin use/dyslipidemia, age, blood pressure level, and diabetes. Non‐adherence to hypertension guidelines was evident as 1364 patients classified from moderate to very high risk received monotherapy. Of these, 496 patients had grade 2 or 3 hypertension. Additionally, 88 patients received the undesirable combination of ACEi + ARB, and 267 (15.9%) type 2 diabetes mellitus (T2DM) patients did not receive RAS‐blockers (146 on monotherapy). The findings reveal a trend toward utilizing ARBs, CCBs, and combination tablets, indicating improved adherence to guidelines. However, a significant number of patients did not receive appropriate treatment, highlighting areas for improvement in prescription practices.

Abstract Cryptococcal meningitis is a high-mortality infection. Adding 5-fluorocytosine (5-FC) to its treatment improves outcomes, but resistance to 5-FC presents a significant challenge. We conducted whole-genome sequencing on seven C. neoformans isolates with varying 5-FC susceptibility, along with proteomic and in silico analyses. Our findings indicate that mutations in genes of the pyrimidine salvage pathway are responsible for 5-FC resistance. Specifically, we identified an E64G missense mutation in the FUR1 gene, a large deletion in the FCY1 gene, and a point mutation in FCY1 leading to a truncated protein. The proteomic data indicated that these mutations resulted in the absence or reduction of crucial enzymes in resistant isolates. Genetic transformations confirmed the association between these mutations and 5-FC resistance. Resistance to 5-FC can develop during treatment and is closely tied to mutations in key metabolic enzymes. Understanding in vivo resistance development is crucial for combating resistance and enhancing patient outcomes.

Background: Participation in cardiac rehabilitation (CR) reduces risk of cardiovascular mortality, improves functional capacity and enhances quality of life in patients with coronary artery disease (CAD). eHealth-based CR can increase participation rates, but research into effective components is necessary. The objective of this systematic review was to identify effective behavior change techniques (BCTs) used in eHealth-based CR interventions. Methods: A search of four databases (CINAHL, PubMed, PsychINFO, and MEDLINE) was conducted until January 10, 2023. Randomized controlled trials investigating eHealth-based interventions for patients with CAD were included. Risk of bias was assessed using the Effective Public Healthcare Practice Project tool. BCTs were coded following the Behavior Change Taxonomy. A best-evidence synthesis was conducted to determine the effectiveness of BCTs, with ratings ranging from A (strong evidence indicating either a positive effect (+) or no effect (-)) to D (no data collected). Results: A total of 88 studies (25,007 participants) met the eligibility criteria. The interventions in these studies used 31 different BCTs. The most common BCTs were instructions on how to perform the behavior (k = 86), social support (k = 69) and information about health consequences (k = 56). The evidence for action planning was rated as A+ for medication adherence and diet. Conversely, for systematically decreasing the number of prompts/cues sent during an intervention, the evidence was rated as A- for physical activity, medication adherence and smoking cessation. The evidence for feedback on behavior was rated as A+ for medication adherence and A- for smoking cessation. Conclusions: Action planning is effective as a BCT in eHealth-based CR, whereas reducing prompts/cues is not. Feedback on behavior may, depending on the behavior targeted, exert both positive and no effect, suggesting that BCT-behavior matching is important to optimize effectiveness of eHealth-based CR.

ABSTRACT Monogenean parasites belonging to Quadriacanthus have never been reported from Clarias ngamensis, but are commonly found on congeners. In view of the specificity of monogeneans, the genetic proximity and the co-occurrence of C. ngamensis and C. gariepinus in Africa, it is hypothesised that a parasitological survey of C. ngamensis will yield several species of Quadriacanthus as its congeneric host C. gariepinus harbours these parasites. The present study aims to explore the monogenean parasites from the gills of C. ngamensis in the Upper Congo Basin by (i) inventorying their diversity and providing their morphological characterisation, and (ii) analysing their infection parameters. Monogeneans were isolated from the gills, and identified based on morphological analysis of genital and haptoral sclerotized parts. Seven parasite species belonging to Quadriacanthus were found. Among them, two species, Q. aegypticus and Q. allobychowskiella, are both newly reported in the Upper Congo Basin and newly recorded on C. ngamensis. Quadriacanthus amakaliae sp. nov., Q. halajiani sp. nov., Q. domatanai sp. nov., Q. lubandaensis sp. nov., and Q. curvicirrus sp. nov. are new to science and described herein. The most prevalent parasite species was Q. curvicirrus sp. nov. in the Luapula River (prevalence = 66.7%) with mean infection intensity of 6.3 ± 5.8. Quadriacanthus allobychowskiella and Q. aegypticus are known parasites from C. gariepinus and their presence on C. ngamensis is probably the result of lateral transfers. The discovery of five new species from C. ngamensis highlights the parasite diversity still to be explored in the region.

Summary: The genome of esophageal adenocarcinoma (EAC) is highly unstable and might evolve over time. Here, we track karyotype evolution in EACs in response to treatment and upon recurrence through multi-region and longitudinal analysis. To this end, we introduce L-PAC (low-purity inference of absolute copy-number alterations [CNAs]), a bio-informatics technique that allows inference of absolute CNAs of low-purity samples by leveraging the information of high-purity samples from the same cancer. Quantitative analysis of matched absolute CNAs reveals that the amount of karyotype evolution induced by chemoradiotherapy (CRT) is predictive for early recurrence and depends on the initial level of karyotype intra-tumor heterogeneity. We observe that CNAs acquired in response to CRT are partially reversed back to the initial state upon recurrence. Hence, CRT alters the fitness landscape to which tumors can adjust by adapting their karyotype. Together, our results indicate that karyotype plasticity contributes to the therapy resistance of EACs.

Abstract Background Persistent infections with high-risk human papillomavirus (hrHPV) can cause cervical squamous intraepithelial lesions (SIL) that may progress to cancer. The cervicovaginal microbiome (CVM) correlates with SIL, but the temporal composition of the CVM after hrHPV infections has not been fully clarified. Methods To determine the association between the CVM composition and infection outcome, we applied high-resolution microbiome profiling using the circular probe-based RNA sequencing technology on a longitudinal cohort of cervical smears obtained from 141 hrHPV DNA-positive women with normal cytology at first visit, of whom 51 were diagnosed by cytology with SIL six months later. Results Here we show that women with a microbial community characterized by low diversity and high Lactobacillus crispatus abundance at both visits exhibit low risk to SIL development, while women with a microbial community characterized by high diversity and Lactobacillus depletion at first visit have a higher risk of developing SIL. At the level of individual species, we observed that a high abundance for Gardnerella vaginalis and Atopobium vaginae at both visits associate with SIL outcomes. These species together with Dialister micraerophilus showed a moderate discriminatory power for hrHPV infection progression. Conclusions Our results suggest that the CVM can potentially be used as a biomarker for cervical disease and SIL development after hrHPV infection diagnosis with implications on cervical cancer prevention strategies and treatment of SIL.

In the Upper Congo Basin, there are few records of monopisthocotylan parasites from clariids. More surveys of clariid fishes can lead to the discovery of multiple monopisthocotylan species that are new to the region or new to science. We aimed to investigate the monopisthocotylan parasite fauna belonging to Quadriacanthus of five clariid fishes in the Upper Congo Basin by (i) inventorying the species composition and providing the description of new species when necessary, and (ii) analysing their infection parameters. Clarias ngamensis, C. stappersii, C. buthupogon, C. gariepinus and C. theodorae were purchased from fishermen in the Lufira, Lubumbashi and Kafubu rivers in the Upper Congo Basin. Monopisthocotylans were mounted on glass slides with ammonium picrate-glycerin for identification based on morphological analysis of genital and haptoral sclerotised parts. Eight species, namely Q. aegypticus, Q. allobychowskiella, Q. amakaliae, Q. domatanai, Q. halajiani, Q. kalomboi n. sp., Q. bassocki n. sp. and Q. shigoleyae n. sp. are reported. The most prevalent parasite species was Q. amakaliae on C. stappersii (prevalence 60%) with a mean infection intensity of 9.8 ± 7.7. We report C. stappersii and C. buthupogon as new hosts for species of Quadriacanthus as well as three new parasite species for science and new localities for the first five parasite species mentioned above. The new records and the description of Q. kalomboi n. sp., Q. bassocki n. sp. and Q. shigoleyae n. sp. increase the knowledge of the diversity of monopisthocotylans in this region.

Effective communication is a crucial objective for business leaders, educators, and politicians alike. Achieving impactful communication involves not only the selection of appropriate words but also proficiency in their delivery. Previous research has frequently examined linguistic, affective linguistic, and paralinguistic features in isolation, thereby overlooking their cumulative impact over time. This study addresses this gap by utilizing a machine learning approach to analyze the dynamic interplay between affective linguistic and paralinguistic features across various episodes of online podcasts. Furthermore, this research incorporates an analysis of gender disparities, acknowledging the dimorphic nature of language and speech across genders. Our findings suggest that accounting for gender when examining the dynamic interactions between affective linguistic and paralinguistic features over time, known as emotional volatility, significantly improves the explanatory power of variations in audience engagement compared to analyses that consider these variables separately.

Objective: To evaluate paroxysmal atrial fibrillation (AF) prevalence in Indian adults who completed 24-Hour Holter monitoring. Methods: A total of 23,847 patients (36.9 % women) were analyzed for AF duration using a software algorithm. Results: AF was diagnosed in 4153 (17.4 %) patients with a median AF duration of 13 min and 55 s. Conclusion: AF prevalence was high and largely untreated. The short duration of AF episodes indicates a low likelihood of detection during clinical visits, highlighting its potential underestimation in Indian healthcare.

Across the western United States, forests are changing rapidly, with uncertain impacts on snowmelt water resources. Snow partitioning is controlled by forest effects on interception, radiation, and sublimation. Yet, models often lack snow measurements with sufficiently high spatial and temporal resolution across gradients of forest structure to accurately represent these fine-scale processes. Here, we utilize four Snowtography stations in Arizona, in the lower Colorado River Basin, with daily measurements over 3–5 years at ~110 positions distributed across gradients of forest structure resulting from wildfires and mechanical thinning. We combine Snowtography with lidar snapshots of forest and snow to train a high-resolution snow model and run it for 6 years to quantify how forest structure regulates snowpack and snowmelt. These study sites represent a climate gradient from lower/warmer ephemeral snowpack (~2,100 m asl) to higher/colder seasonal snowpack (~2,800 m asl). Forest cover reduced snowpack and snowmelt through canopy sublimation. Forest advanced snowmelt timing at lower/warmer sites but delayed it at higher/colder sites. Within canopy gaps, shaded cool edges had the greatest peak snow water equivalent (SWE). Surprisingly, sunny/warm gap edges produced more snowmelt than cool edges, because high radiation melted snow quickly, reducing exposure to sublimation. Therefore, peak SWE is not an ideal proxy for snowmelt volume from ephemeral snowpacks, which are becoming more prevalent due to warming. The results imply that forest management can influence the amount and timing of snowmelt, and that there may be decision trade-offs between enhancing forest resilience through delayed snowmelt and maximizing snowmelt volumes for downstream water resources.

Summary: Gustatory receptors (GRs) are critical for insect chemosensation and are potential targets for controlling pests and disease vectors, making their structural investigation a vital step toward such applications. We present structures of Bombyx mori Gr9 (BmGr9), a fructose-gated cation channel, in agonist-free and fructose-bound states. BmGr9 forms a tetramer similar to distantly related insect odorant receptors (ORs). Upon fructose binding, BmGr9’s channel gate opens through helix S7b movements. In contrast to ORs, BmGr9’s ligand-binding pocket, shaped by a kinked helix S4 and a shorter extracellular S3-S4 loop, is larger and solvent accessible in both agonist-free and fructose-bound states. Also, unlike ORs, fructose binding by BmGr9 involves helix S5 and a pocket lined with aromatic and polar residues. Structure-based sequence alignments reveal distinct patterns of ligand-binding pocket residue conservation in GR subfamilies associated with different ligand classes. These data provide insight into the molecular basis of GR ligand specificity and function.

During a parasitological survey carried out between May and August 2022 in the River Nyando, Lake Victoria Basin, a single species of Rhabdochona Railliet, 1916 (Nematoda: Rhabdochonidae) was recorded from the intestine of the Rippon barbel, Labeobarbus altianalis (Boulenger, 1900) (Cyprinidae). Based on light microscopy (LM), scanning electron microscopy (SEM) and DNA analyses the parasite was identified as Rhabdochona (Rhabdochona) gendrei Campana-Rouget, 1961. Light microscopy, SEM and DNA studies on this rhabdochonid resulted in a detailed redescription of the adult male and female. The following additional taxonomic features are described in the male: 14 anterior prostomal teeth; 12 pairs of preanal papillae: 11 subventral and one lateral; six pairs of postanal papillae: five subventral and one lateral, with the latter pair at the level of first subventral pairs when counted from the cloacal aperture. For the female: 14 anterior prostomal teeth and the size and absence of superficial structures on fully mature (larvated) eggs dissected out of the nematode body. Specimens of R. gendrei were genetically distinct in the 28S rRNA and cytochrome c oxidase subunit 1 (cox1) mitochondrial gene regions from known species of Rhabdochona. This is the first study that provides genetic data for a species of Rhabdochona from Africa, the first SEM of R. gendrei, and the first report of this parasite from Kenya. The molecular and SEM data reported herein provide a useful point of reference for future studies on Rhadochona in Africa.

Background: Previous research has shown that the Serum Amyloid A (SAA) protein family is intricately involved in inflammatory signaling and various disease pathologies. We have previously demonstrated that SAA is associated with increased colitis disease severity and the promotion of tumorigenesis. However, the specific role of SAA proteins in breast cancer pathology remains unclear. Therefore, we investigated the role of systemic SAA1 and SAA2 (SAA1/2) in a triple-negative breast cancer mouse model. Methods: Syngeneic breast tumors were established in wild-type mice, and mice lacking the SAA1/2 (SAADKO). Subsequently, tumor volume was monitored, species survival determined, the inflammatory profiles of mice assessed with a multiplex assay, and tumor molecular biology and histology characterized with Western blotting and H&E histological staining. Results: WT tumor-bearing mice had increased levels of plasma SAA compared to wild-type control mice, while SAADKO control and tumor-bearing mice presented with lower levels of SAA in their plasma. SAADKO tumor-bearing mice also displayed significantly lower concentrations of systemic inflammatory markers. Tumors from SAADKO mice overall had lower levels of SAA compared to tumors from wild-type mice, decreased apoptosis and inflammasome signaling, and little to no tumor necrosis. Conclusions: We demonstrated that systemic SAA1/2 stimulates the activation of the NLRP3 inflammasome in breast tumors, leading to the production of pro-inflammatory cytokines. This, in turn, promoted apoptosis and tumor necrosis but did not significantly impact tumor growth or histological grading.

Background: Cardiac rehabilitation (CR) reduces recurrent cardiac events and mortality in patients with cardiovascular diseases (CVD). Innovative eHealth methods can facilitate CR uptake and effectiveness by addressing barriers associated with clinic-based rehabilitation. Tailoring eHealth-based CR to patient preferences is needed to further enhance CR. Purpose: To identify preferred behavior change techniques (BCTs) as well as barriers and facilitators for the different health behaviors targeted in eHealth-based CR among patients who have been referred to CR. Methods: Thirty-nine patients were interviewed in nine focus groups in The Netherlands, Germany, and Spain. A thematic analysis, using a combined deductive and inductive approach to coding, was conducted to identify BCTs and barriers and facilitators to behavior change. Behaviors under investigation included physical activity, medication adherence, eating a cardiac healthy-diet, stress reduction and smoking cessation. Results: The perceived helpfulness of BCTs depended on the specific behavior targeted. Common barriers were negative emotional state and physical limitations. A desire to feel physically or mentally well and having experienced a cardiac life event were the most common facilitators across health behaviors. Specific BCTs, barriers and facilitators were found for each of the health behavior. Conclusions: Behavior change techniques that patients preferred for each health behavior targeted in eHealth-based CR were identified. A negative emotional state, experiencing a life event, and improving physical functioning are important barriers and facilitators in multiple behaviors targeted in eHealth-based CR programs. Additional tailoring of interventions to patient preferences for BCTs and patient-specific barriers and facilitators per health behavior could lead to further improvement of eHealth-based CR.

Aim: Out-of-hospital cardiac arrest is a major health problem, and the overall survival rate is low (4.6%–16.4%). The initiation of the current chain of survival depends on the presence of a witness of the cardiac arrest, which is not present in 29.7%–63.4% of the cases. Furthermore, a delay in starting this chain is common in witnessed out-of-hospital cardiac arrest. This project aims to reduce morbidity and mortality due to out-of-hospital cardiac arrest by developing a smartwatch-based solution to expedite the chain of survival in the case of (un)witnessed out-of-hospital cardiac arrest. Methods: Within the ‘Beating Cardiac Arrest’ project, we aim to develop a demonstrator product that detects out-of-hospital cardiac arrest using photoplethysmography and accelerometer analysis, and autonomously alerts emergency medical services. A target group study will be performed to determine who benefits the most from this product. Furthermore, several clinical studies will be conducted to capture or simulate data on out-of-hospital cardiac arrest cases, as to develop detection algorithms and validate their diagnostic performance. For this, the product will be worn by patients at high risk for out-of-hospital cardiac arrest, by volunteers who will temporarily interrupt blood flow in their arm by inflating a blood pressure cuff, and by patients who undergo cardiac electrophysiologic and implantable cardioverter defibrillator testing procedures. Moreover, studies on psychosocial and ethical acceptability will be conducted, consisting of surveys, focus groups, and interviews. These studies will focus on end-user preferences and needs, to ensure that important individual and societal values are respected in the design process.

ABSTRACT The emerging prevalence of extended-spectrum beta-lactamase (ESBL) producing Enterobacterales has implications for the empirical treatment of common infections, such as complicated urinary tract infections. To provide adequate treatment, while avoiding empirical therapy with last resort carbapenems, rapid identification of ESBL-containing pathogens is desirable. Routine urine samples were collected between February and July 2021 in two Dutch clinical medical microbiology laboratories according to a predefined list containing certain culture characteristics. All urine samples were screened for the presence of ESBL genes (bla CTX-M2, bla CTX-M14, and bla CTX-M15) with random-access quantitative PCR (qPCR) using the Cepheid GeneXpert ESBL-ampC prototype assay. The qPCR and microbiological culture results were compared. After the calculation of the sensitivity and specificity, discrepancies were investigated by whole-genome sequencing. In total, 276 urine samples were available for ESBL analysis (94 ESBL culture positive and 182 ESBL culture negative). The sensitivity and specificity for detection of ESBL genes were 90.4% and 98.4%, respectively. In nine samples (9.6%), no ESBL genes were detected with GeneXpert, while in the microbiological culture, an ESBL-positive organism was isolated. This was mainly explained by non-GeneXpert ESBL genes: bla SHV-family (n = 6; 75.0%), bla TEM-family (n = 1; 12.5%), and bla SRT-family (n = 1; 12.5%). The positive and negative predictive values in a hypothetical clinical scenario with a 15% ESBL prevalence were 0.91 and 0.98, respectively. Regarding ampC, the specificity appears to be satisfactory, but the sensitivity is low. The Cepheid GeneXpert ESBL assay could be beneficial for the fast and accurate detection of ESBL genes in regions where the epidemiology of ESBL genes coincides with the targets in the panel. IMPORTANCE Early identification of complicated urinary tract infections caused by ESBL-producing Enterobacterales has the potential to limit the use of carbapenems to those patients without alternative antibiotic options and avoid the empirical use of carbapenems in patients without ESBL-producing bacteria. The purpose for such a test will differ by setting and ESBL prevalence rates. Countries with low ESBL rates and cephalosporins as empiric treatment (e.g., The Netherlands) will need a rule-in test to decide to use carbapenems, while countries with high ESBL rates and empiric carbapenem treatment will need a rule-out test for ESBLs to de-escalate therapy early. Anyway, such as a test would—at least theoretically—improve patient care and reduce selective pressure for the emergence of carbapenem resistance.

Pre-fire environmental conditions play a critical role in wildfire severity. This study investigated the impact of pre-fire forest conditions on burn severity as a result of the 2020 Bighorn Fire in the Santa Catalina Mountains in Arizona. Using a stepwise regression model and remotely sensed data from Landsat 8 and LiDAR, we analyzed the effects of structural and functional vegetation traits and environmental factors on burn severity. This analysis revealed that the difference normalized burn ratio (dNBR) was a more reliable indicator of burn severity compared to the relative dNBR (RdNBR). Stepwise regression identified pre-fire normalized difference vegetation index (NDVI), canopy cover, and tree density as significant variables across all land cover types that explained burn severity, suggesting that denser areas with higher vegetation greenness experienced more severe burns. Interestingly, residuals between the actual and estimated dNBR were lower in herbaceous zones compared to denser forested areas at similar elevations, suggesting potentially more predictable burn severity in open areas. Spatial analysis using Geary’s C statistics further revealed a strong negative autocorrelation: areas with high burn severity tended to be clustered, with lower severity areas interspersed. Overall, this study demonstrates the potential of readily available remote sensing data to predict potential burn severity values before a fire event, providing valuable information for forest managers to develop strategies for mitigating future wildfire damage.

Background General practitioners (GPs) are increasingly confronted with people with both mild intellectual disability (MID) and mental health (MH) problems. Little is known about the type of MH problems for which people with MID visit their GP and the care provided.Objectives To identify the type and prevalence of MH disorders and MH-related complaints in people with MID in primary care and care provided, compared to people without ID.Methods By linking the Netherlands Institute for Health Services Research’s primary care databases, comprising electronic health records, with Statistic Netherlands’ social services and chronic care databases, we identified 11,887 people with MID. In this four-year retrospective study, MH-related International Classification of Primary Care (ICPC) codes and care characteristics were compared between people with MID and without ID.Results Of the people with MID, 48.8% had MH problems recorded vs. 30.4% of the people without ID, with significant differences in substance abuse, suicide attempts, and psychosis. Of the MID group, 80.3% were not registered by their GP with the ICPC code mental retardation. GPs provided more care to people with MID and MH problems than people without ID but with MH-problems regarding consultations (median 6.4 vs. 4.0 per year) and variety of prescribed medications (median 2.7 vs. 2.0 per year).Conclusion In primary care, the prevalence of MH problems and care provided is high in people with MID. To improve primary mental healthcare for this group, it is essential to increase GPs’ awareness and knowledge on the combination of MID and MH.

Myeloid cells contribute to the tumor microenvironment of thyroid cancers, but their functional relevance is lesser known. Here authors show that myeloid cells infiltrating non-medullary thyroid carcinomas upregulate their antigen presentation-related genes, release less cytokines and over-produce reactive oxygen species, with transcriptional changes already present in extra-tumoral myeloid cells.

Background: Surgical treatment of proximal femur fractures is complicated by postoperative delirium in about one-third of patients. Pain and opioid consumption are modifiable factors that may influence the incidence of delirium.1 An intrathecal injection of morphine may lead to a reduction in postoperative pain and reduced systemic opioid consumption. In current practice, the addition of morphine to intrathecal anaesthesia is commonly used but depends on the anaesthesiologist's preference. Recently, a retrospective study found that intrathecal morphine was independently associated with a lower incidence of delirium. However, this has to be confirmed in a prospective, randomised study. We hypothesise that using intrathecal morphine reduces postoperative pain and opioid consumption during the first 48 h after surgery and reduces the incidence of delirium during hospital admission. We also seek additional evidence of the association between neuronal injury (delirium) and neurofilament light in serum of patients with proximal femur fractures. Objective: The primary objective is to compare the incidence of delirium. The secondary objectives are to compare pain scores, systemic opioid consumption, and (opioid-related) side-effects. The tertiary objective is to test the association between intrathecal morphine and neurofilament light as a marker of neuronal injury. Study design: A double-blind, randomised, placebo-controlled intervention study is proposed. Study population: All patients with a proximal femur fracture who are scheduled for surgery under spinal anaesthesia. Intervention: The intervention is the addition of morphine 100 μg to the intrathecal injection for spinal anaesthesia. The intervention group will receive a mixture of bupivacaine 10 mg and morphine 100 μg. The control group will receive bupivacaine 10 mg. Clinical trial registration: EU Clinical Trials Register: EudraCT number 2020-002143-27.

Abstract The cervicovaginal microbiome (CVM) correlates with women’s cervical health, and variations in its composition are associated with high-risk human papillomavirus (hrHPV) infection outcomes. Cervicovaginal microbes have been grouped into five community state types (CSTs) based on microbial community composition and abundance. However, studying the impact of CSTs in health and disease is challenging because the current sequencing technologies have limited confident discrimination between closely related and yet functionally different bacterial species. Circular probe-based RNA sequencing (ciRNAseq) achieves high-resolution microbiome profiling and therefore provides in-depth and unambiguous knowledge about the composition of the CVM. Based on ciRNAseq profiling of a large cohort of cervical smears (n = 541), we here define subgroups of CSTs I, III, and IV based on intra-CST differences with respect to abundances of Lactobacillus acidophilus (CSTs I-A vs. I-B and CSTs III-A vs. III-B), Lactobacillus iners (CSTs I-A vs. I-B and CSTs III-A vs. III-B), and Megasphaera genomosp type 1 (CSTs IV-A vs. IV-B). Our results further support the existence of subgroups of CST IV-C that are dominant for non-Lactobacillus species and have intermediate microbial diversity. We also show that CST V is associated with uninfected conditions, and CST IV-A associates with hrHPV-induced cervical disease. In conclusion, we characterized new subdivisions of cervicovaginal CSTs, which may further advance our understanding of women’s cervical health and hrHPV-related progression to disease.

Abstract Hypertension guidelines recommend measuring blood pressure (BP) in both arms at least once. However, this is seldom done due to uncertainties regarding measurement procedure and the implications of finding a clinically important inter‐arm BP difference (IAD). This study aimed to provide insight into the prevalence of clinically important IADs in a large Indian primary care cohort. A number of 134 678 (37% female) unselected Indian primary care participants, mean age 45.2 (SD 11.9) years, had BP measured in both arms using a standardized, triplicate, automated simultaneous measurement method (Microlife WatchBP Office Afib). On average, there were clinically minor differences in right and left arm BP values: systolic BP 134.4 vs 134.2 mmHg (p

Abstract Background Patient decision aids (PtDAs) support patients and clinicians in shared decision-making (SDM). Real-world outcome information may improve patients’ risk perception, and help patients make decisions congruent with their expectations and values. Our aim was to develop an online PtDA to support kidney failure treatment modality decision-making, that: 1) provides patients with real-world outcome information, and 2) facilitates SDM in clinical practice. Methods The International Patient Decision Aids Standards (IPDAS) development process model was complemented with a user-centred and convergent mixed-methods approach. Rapid prototyping was used to develop the PtDA with a multidisciplinary steering group in an iterative process of co-creation. The results of an exploratory evidence review and a needs-assessment among patients, caregivers, and clinicians were used to develop the PtDA. Seven Dutch teaching hospitals and two national Dutch outcome registries provided real-world data on selected outcomes for all kidney failure treatment modalities. Alpha and beta testing were performed to assess the prototype and finalise development. An implementation strategy was developed to guide implementation of the PtDA in clinical practice. Results The ‘Kidney Failure Decision Aid’ consists of three components designed to help patients and clinicians engage in SDM: 1) a paper hand-out sheet, 2) an interactive website, and 3) a personal summary sheet. A ‘patients-like-me’ infographic was developed to visualise survival probabilities for each treatment modality on the website. Other treatment outcomes were incorporated as event rates (e.g. hospitalisation rates) or explained in text (e.g. the flexibility of each treatment modality). No major revisions were needed after alpha and beta testing. During beta testing, some patients ignored the survival probabilities because they considered these too confronting. Nonetheless, patients agreed that every patient has the right to choose whether they want to view this information. Patients and clinicians believed that the PtDA would help patients make informed decisions, and that it would support values- and preferences-based decision-making. Implementation of the PtDA has started in October 2020. Conclusions The ‘Kidney Failure Decision Aid’ was designed to facilitate SDM in clinical practice and contains real-world outcome information on all kidney failure treatment modalities. It is currently being investigated for its effects on SDM in a clinical trial.

Whole genome sequencing (WGS) is widely used for outbreak analysis of bacteriology and virology but is scarcely used in mycology. Here, we used WGS for genotyping Aspergillus fumigatus isolates from a potential Aspergillus outbreak in an intensive care unit (ICU) during construction work. After detecting the outbreak, fungal cultures were performed on all surveillance and/or patient respiratory samples. Environmental samples were obtained throughout the ICU. WGS was performed on 30 isolates, of which six patient samples and four environmental samples were related to the outbreak, and twenty samples were unrelated, using the Illumina NextSeq 550. A SNP-based phylogenetic tree was created from outbreak samples and unrelated samples. Comparative analysis (WGS and short tandem repeats (STRs), microsatellite loci analysis) showed that none of the strains were related to each other. The lack of genetic similarity suggests the accumulation of Aspergillus spores in the hospital environment, rather than a single source that supported growth and reproduction of Aspergillus fumigatus. This supports the hypothesis that the Aspergillus outbreak was likely caused by release of Aspergillus fumigatus spores during construction work. Indeed, no new Aspergillus cases were observed in the ICU after cessation of construction. This study demonstrates that WGS is a suitable technique for examining inter-strain relatedness of Aspergillus fumigatus in the setting of an outbreak investigation.

Abstract Background Because most cervical cancers are caused by high-risk human papillomaviruses (hrHPVs), cervical cancer prevention programs increasingly employ hrHPV testing as a primary test. The high sensitivity of HPV tests is accompanied by low specificity, resulting in high rates of overdiagnosis and overtreatment. Targeted circular probe-based RNA next generation sequencing (ciRNAseq) allows for the quantitative detection of RNAs of interest with high sequencing depth. Here, we examined the potential of ciRNAseq-testing on cervical scrapes to identify hrHPV-positive women at risk of having or developing high-grade cervical intraepithelial neoplasia (CIN). Methods We performed ciRNAseq on 610 cervical scrapes from the Dutch cervical cancer screening program to detect gene expression from 15 hrHPV genotypes and from 429 human genes. Differentially expressed hrHPV- and host genes in scrapes from women with outcome “no CIN” or “CIN2+” were identified and a model was built to distinguish these groups. Results Apart from increasing percentages of hrHPV oncogene expression from “no CIN” to high-grade cytology/histology, we identified genes involved in cell cycle regulation, tyrosine kinase signaling pathways, immune suppression, and DNA repair being expressed at significantly higher levels in scrapes with high-grade cytology and histology. Machine learning using random forest on all the expression data resulted in a model that detected ‘no CIN’ versus CIN2+ in an independent data set with sensitivity and specificity of respectively 85 ± 8% and 72 ± 13%. Conclusions CiRNAseq on exfoliated cells in cervical scrapes measures hrHPV-(onco)gene expression and host gene expression in one single assay and in the process identifies HPV genotype. By combining these data and applying machine learning protocols, the risk of CIN can be calculated. Because ciRNAseq can be performed in high-throughput, making it cost-effective, it can be a promising screening technology to stratify women at risk of CIN2+. Further increasing specificity by model improvement in larger cohorts is warranted.

Abstract In recent years, cardiovascular immuno-imaging by positron emission tomography (PET) has undergone tremendous progress in preclinical settings. Clinically, two approved PET tracers hold great potential for inflammation imaging in cardiovascular patients, namely FDG and DOTATATE. While the former is a widely applied metabolic tracer, DOTATATE is a relatively new PET tracer targeting the somatostatin receptor 2 (SST2). In the current study, we performed a detailed, head-to-head comparison of DOTATATE-based radiotracers and [18F]F-FDG in mouse and rabbit models of cardiovascular inflammation. For mouse experiments, we labeled DOTATATE with the long-lived isotope [64Cu]Cu to enable studying the tracer’s mode of action by complementing in vivo PET/CT experiments with thorough ex vivo immunological analyses. For translational PET/MRI rabbit studies, we employed the more widely clinically used [68Ga]Ga-labeled DOTATATE, which was approved by the FDA in 2016. DOTATATE’s pharmacokinetics and timed biodistribution were determined in control and atherosclerotic mice and rabbits by ex vivo gamma counting of blood and organs. Additionally, we performed in vivo PET/CT experiments in mice with atherosclerosis, mice subjected to myocardial infarction and control animals, using both [64Cu]Cu-DOTATATE and [18F]F-FDG. To evaluate differences in the tracers’ cellular specificity, we performed ensuing ex vivo flow cytometry and gamma counting. In mice subjected to myocardial infarction, in vivo [64Cu]Cu-DOTATATE PET showed higher differential uptake between infarcted (SUVmax 1.3, IQR, 1.2–1.4, N = 4) and remote myocardium (SUVmax 0.7, IQR, 0.5–0.8, N = 4, p = 0.0286), and with respect to controls (SUVmax 0.6, IQR, 0.5–0.7, N = 4, p = 0.0286), than [18F]F-FDG PET. In atherosclerotic mice, [64Cu]Cu-DOTATATE PET aortic signal, but not [18F]F-FDG PET, was higher compared to controls (SUVmax 1.1, IQR, 0.9–1.3 and 0.5, IQR, 0.5–0.6, respectively, N = 4, p = 0.0286). In both models, [64Cu]Cu-DOTATATE demonstrated preferential accumulation in macrophages with respect to other myeloid cells, while [18F]F-FDG was taken up by macrophages and other leukocytes. In a translational PET/MRI study in atherosclerotic rabbits, we then compared [68Ga]Ga-DOTATATE and [18F]F-FDG for the assessment of aortic inflammation, combined with ex vivo radiometric assays and near-infrared imaging of macrophage burden. Rabbit experiments showed significantly higher aortic accumulation of both [68Ga]Ga-DOTATATE and [18F]F-FDG in atherosclerotic (SUVmax 0.415, IQR, 0.338–0.499, N = 32 and 0.446, IQR, 0.387–0.536, N = 27, respectively) compared to control animals (SUVmax 0.253, IQR, 0.197–0.285, p = 0.0002, N = 10 and 0.349, IQR, 0.299–0.423, p = 0.0159, N = 11, respectively). In conclusion, we present a detailed, head-to-head comparison of the novel SST2-specific tracer DOTATATE and the validated metabolic tracer [18F]F-FDG for the evaluation of inflammation in small animal models of cardiovascular disease. Our results support further investigations on the use of DOTATATE to assess cardiovascular inflammation as a complementary readout to the widely used [18F]F-FDG.

ABSTRACT The detection and accurate identification of bacterial species in clinical samples are crucial for diagnosis and appropriate antibiotic treatment. To date, sequencing of the 16S rRNA gene has been widely used as a complementary molecular approach when identification by culture fails. The accuracy and sensitivity of this method are highly affected by the selection of the 16S rRNA gene region targeted. In this study, we assessed the clinical utility of 16S rRNA reverse complement PCR (16S RC-PCR), a novel method based on next-generation sequencing (NGS), for the identification of bacterial species. We investigated the performance of 16S RC-PCR on 11 bacterial isolates, 2 polymicrobial community samples, and 59 clinical samples from patients suspected of having a bacterial infection. The results were compared to culture results, if available, and to the results of Sanger sequencing of the 16S rRNA gene (16S Sanger sequencing). By 16S RC-PCR, all bacterial isolates were accurately identified to the species level. Furthermore, in culture-negative clinical samples, the rate of identification increased from 17.1% (7/41) to 46.3% (19/41) when comparing 16S Sanger sequencing to 16S RC-PCR. We conclude that the use of 16S RC-PCR in the clinical setting leads to an increased sensitivity of detection of bacterial pathogens, resulting in a higher number of diagnosed bacterial infections, and thereby can improve patient care. IMPORTANCE The identification of the causative infectious pathogen in patients suspected of having a bacterial infection is essential for diagnosis and the start of appropriate treatment. Over the past 2 decades, molecular diagnostics have improved the ability to detect and identify bacteria. However, novel techniques that can accurately detect and identify bacteria in clinical samples and that can be implemented in clinical diagnostics are needed. Here, we demonstrate the clinical utility of bacterial identification in clinical samples by a novel method called 16S RC-PCR. Using 16S RC-PCR, we reveal a significant increase in the number of clinical samples in which a potentially clinically relevant pathogen is identified compared to the commonly used 16S Sanger method. Moreover, RC-PCR allows automation and is well suited for implementation in a diagnostic laboratory. In conclusion, the implementation of this method as a diagnostic tool is expected to result in an increased number of diagnosed bacterial infections, and in combination with adequate treatment, this could improve clinical outcomes for patients.

ABSTRACT The opportunistic fungal pathogen Aspergillus fumigatus utilizes two high-affinity iron uptake mechanisms, termed reductive iron assimilation (RIA) and siderophore-mediated iron acquisition (SIA). The latter has been shown to be crucial for virulence of this fungus and is a target for development of novel strategies for diagnosis and treatment of fungal infections. So far, research on SIA in this mold focused mainly on the hyphal stage, revealing the importance of extracellular fusarinine-type siderophores in iron acquisition as well as of the siderophore ferricrocin in intracellular iron handling. The current study aimed to characterize iron acquisition during germination. High expression of genes involved in biosynthesis and uptake of ferricrocin in conidia and during germination, independent of iron availability, suggested a role of ferricrocin in iron acquisition during germination. In agreement, (i) bioassays indicated secretion of ferricrocin during growth on solid media during both iron sufficiency and limitation, (ii) ferricrocin was identified in the supernatant of conidia germinating in liquid media during both iron sufficiency and limitation, (iii) in contrast to mutants lacking all siderophores, mutants synthesizing ferricrocin but lacking fusarinine-type siderophores were able to grow under iron limitation in the absence of RIA, and (iv) genetic inactivation of the ferricrocin transporter Sit1 decreased germination in the absence of RIA. Taken together, this study revealed that ferricrocin has not only an intracellular role but also functions as an extracellular siderophore to support iron acquisition. The iron availability-independent ferricrocin secretion and uptake during early germination indicate developmental, rather than iron regulation. IMPORTANCE Aspergillus fumigatus is one of the most common airborne fungal pathogens for humans. Low-molecular-mass iron chelators, termed siderophores, have been shown to play a central role in iron homeostasis and, consequently, virulence of this mold. Previous studies demonstrated the crucial role of secreted fusarinine-type siderophores, such as triacetylfusarinine C, in iron acquisition, as well as of the ferrichrome-type siderophore ferricrocin in intracellular iron storage and transport. Here, we demonstrate that ferricrocin is also secreted to mediate iron acquisition during germination together with reductive iron assimilation. During early germination, ferricrocin secretion and uptake were not repressed by iron availability, indicating developmental regulation of this iron acquisition system in this growth phase.

Cyanodermella asteris is a fungal endophyte from Aster tataricus, a perennial plant from the northern part of Asia. Here, we demonstrated an interaction of C. asteris with Arabidopsis thaliana, Chinese cabbage, rapeseed, tomato, maize, or sunflower resulting in different phenotypes such as shorter main roots, massive lateral root growth, higher leaf and root biomass, and increased anthocyanin levels. In a variety of cocultivation assays, it was shown that these altered phenotypes are caused by fungal CO2, volatile organic compounds, and soluble compounds, notably astins. Astins A, C, and G induced plant growth when they were individually included in the medium. In return, A. thaliana stimulates the fungal astin C production during cocultivation. Taken together, our results indicate a bilateral interaction between the fungus and the plant. A stress response in plants is induced by fungal metabolites while plant stress hormones induced astin C production of the fungus. Interestingly, our results not only show unidirectional influence of the fungus on the plant but also vice versa. The plant is able to influence growth and secondary metabolite production in the endophyte, even when both organisms do not live in close contact, suggesting the involvement of volatile compounds.[Graphic: see text] Copyright © 2021 The Author(s). This is an open access article distributed under the CC BY-NC-ND 4.0 International license.

Abstract Cardiovascular diseases have become the major cause of death in India, but overall awareness is still low. Therefore, the initiative was undertaken to set up health care screening booths at eight airports and one hospital throughout India to increase awareness and to determine cardiovascular risk factors. Participants were screened for hypertension (systolic blood pressure (BP) ≥140 mmHg or diastolic BP≥90 mmHg), diabetes [fasting blood glucose (FBG) level ≥126 or ≥200 mg/dL for random blood glucose (RBG)], and body mass index (BMI). Among 100 107 participants screened (46 ± 13 years; 17% women), prevalence of diabetes was 12 571 (15%), hypertension: 30 345 (33%) and overweight: 61 219 (65%). Diabetes was treated more often than hypertension (44% vs 11%). Hypertension and diabetes prevalence values were relatively high in young obese adults; BMI correlated significantly (p

Background Adults with mild intellectual disability (MID) experience more mental health disorders than the general population. However, mental healthcare may be insufficiently tailored to match their needs. Detailed information is lacking regarding care provided to people with MID in mental health services. Aims To compare mental health disorders and care provided to patients with and without MID in Dutch mental health services, including patients with missing MID status in the service files. Method In this population-based database study, we used a Statistics Netherlands mental health service database, containing health insurance claims of patients who utilised advanced mental health services in 2015–2017. Patients with MID were identified by linking this database with Statistic Netherlands’ social services and long-term care databases. Results We identified 7596 patients with MID, of whom 60.6% had no intellectual disability registration in the service files. Compared with patients without intellectual disability (n = 329 864), they had different profiles of mental health disorders. They received fewer diagnostic (odds ratio 0.71, 95% CI 0.67–0.75) and treatment activities (odds ratio 0.56, 95% CI 0.53–0.59), and required more interprofessional consultations outside of the service (odds ratio 2.06, 95% CI 1.97–2.16), crisis interventions (odds ratio 2.00, 95% CI 1.90–2.10) and mental health-related hospital admissions (odds ratio 1.72, 95% CI 1.63–1.82). Conclusions Patients with MID in mental health services have different profiles of mental health disorders and care than patients without intellectual disability. In particular, fewer diagnostics and treatments are provided, especially in those with MID with no intellectual disability registration, putting patients with MID at risk of undertreatment and poorer mental health outcomes.

Background Compared to the previous cytology-based program, the introduction of primary high-risk human papillomavirus (hrHPV) based screening in 2017 has led to an increased number of referrals. To counter this, triage of hrHPV-positive women in cervical cancer screening can potentially be optimized by taking sociodemographic and lifestyle risk factors for cervical abnormalities into account. Therefore, it is essential to gain knowledge of the views of women (30–60 years) eligible for cervical cancer screening. Objective The main goal of this qualitative study was to gain insight in the aspects that influence acceptability of risk-based triage in cervical cancer screening. Design A focus group study in which participants were recruited via four general medical practices, and purposive sampling was used to maximize heterogeneity with regards to age, education level, and cervical cancer screening experiences. Approach The focus group discussions were transcribed verbatim and analyzed using reflexive thematic analysis. Participants A total of 28 women (average age: 45.2 years) eligible for cervical cancer screening in The Netherlands participated in seven online focus group discussions. Half of the participants was higher educated, and the participants differed in previous cervical cancer screening participation and screening result. Key results In total, 5 main themes and 17 subthemes were identified that determine the acceptability of risk-stratified triage. The main themes are: 1) adequacy of the screening program: an evidence-based program that is able to minimize cancer incidence and reduce unnecessary referrals; 2) personal information (e.g., sensitive topics and stigma); 3) emotional impact: fear and reassurance; 4) communication (e.g., transparency); and 5) autonomy (e.g., prevention). Conclusion The current study highlights several challenges regarding the development and implementation of risk-based triage that need attention in order to be accepted by the target group. These challenges include dealing with sensitive topics and a transparent communication strategy.

Biosensors hold great promise as diagnostic devices that gain the information needed to discern between different types and severities of infection. Accurate diagnostic information allows for appropriate antimicrobial usage, thereby benefiting patient welfare and curbing the development of antimicrobial resistance. With these aims in mind, a nano- and microfiber mesh-based transducer platform for use in aqueous media was developed. When used in an electrochemical cell, this transducer is able to distinguish between different concentrations of phosphate-buffered saline in deionized water using electrochemical impedance spectroscopy. This transducer, when coupled with a biorecognition element, could serve as a new biosensor platform, to be employed as a diagnostic device that could be applied to various biological targets.

Abstract Background The cervicovaginal microbiome (CVM) plays a significant role in women’s cervical health and disease. Microbial alterations at the species level and characteristic community state types (CST) have been associated with acquisition and persistence of high-risk human papillomavirus (hrHPV) infections that may result in progression of cervical lesions to malignancy. Current sequencing methods, especially most commonly used multiplex 16S rRNA gene sequencing, struggle to fully clarify these changes because they generally fail to provide sufficient taxonomic resolution to adequately perform species-level associative studies. To improve CVM species designation, we designed a novel sequencing tool targeting microbes at the species taxonomic rank and examined its potential for profiling the CVM. Results We introduce an accessible and practical circular probe-based RNA sequencing (CiRNAseq) technology with the potential to profile and quantify the CVM. In vitro and in silico validations demonstrate that CiRNAseq can distinctively detect species in a mock mixed microbial environment, with the output data reflecting its ability to estimate microbes’ abundance. Moreover, compared to 16S rRNA gene sequencing, CiRNAseq provides equivalent results but with improved sequencing sensitivity. Analyses of a cohort of cervical smears from hrHPV-negative women versus hrHPV-positive women with high-grade cervical intraepithelial neoplasia confirmed known differences in CST occurring in the CVM of women with hrHPV-induced lesions. The technique also revealed variations in microbial diversity and abundance in the CVM of hrHPV-positive women when compared to hrHPV-negative women. Conclusions CiRNAseq is a promising tool for studying the interplay between the CVM and hrHPV in cervical carcinogenesis. This technology could provide a better understanding of cervicovaginal CST and microbial species during health and disease, prompting the discovery of biomarkers, additional to hrHPV, that can help detect high-grade cervical lesions.

Tracing and reflecting on God should be an integral aspect of practical theological reflection. This article focuses on the Trinity as a way of speaking about the identity of God and how tracing it would inform doing practical theology. Practical theologians reflect in different ways on how they trace God. Using the hermeneutical process is essential in practical theology. Both Browning and Heitink explain the interaction between text and context, while Browning and Osmer opt for an ethical interpretation in engaging with the context. Root and Ward endeavour to focus more on the theological aspects of doing practical theology. Root focuses on Christopraxis and Ward on interpreting the Gospel within the current context. A discussion about the immanent Trinity and the social Trinity illustrates the complexity of tracing God in doing practical theology. The conversation needs to be taken further, by discussing three aspects – ecclesiology, hospitality and lived religion – as markers to illustrate the value of a trinitarian reflection for practical theology. Practical theology could benefit by tracing the Trinity and, in so doing, develop and enhance the theological character of the discipline. Intradisciplinary and/or interdisciplinary implications: The article focuses on the implications of tracing the Trinity in practical theology. A systematic theological reflection informs the discussion and opens a dialogue between the two disciplines. The theological nature of practical theology should be strengthened, enhancing a critical dialogue with social sciences regarding, for example, hospitality and lived religion.

ABSTRACT Methicillin-resistant Staphylococcus aureus (MRSA) clusters are considered epidemic or nonepidemic based on their ability to spread effectively. Successful transmission could be influenced by dehydration tolerance. Current methods for determination of dehydration tolerance lack accuracy. Here, a climate-controlled in vitro dehydration assay using isothermal microcalorimetry (IMC) was developed and linked with mathematical modeling to determine survival of 44 epidemic versus 54 nonepidemic MRSA strains from France, the United Kingdom, and the Netherlands after 1 week of dehydration. For each MRSA strain, the growth parameters time to end of first growth phase (tmax [h]) and maximal exponential growth rate (μm) were deduced from IMC data for 3 experimental replicates, 3 different starting inocula, and before and after dehydration. If the maximal exponential growth rate was within predefined margins (±36% of the mean), a linear relationship between tmax and starting inoculum could be utilized to predict log reduction after dehydration for individual strains. With these criteria, 1,330 of 1,764 heat flow curves (data sets) (75%) could be analyzed to calculate the post-dehydration inoculum size, and thus the log reduction due to dehydration, for 90 of 98 strains (92%). Overall reduction was ~1 log after 1 week. No difference in dehydration tolerance was found between the epidemic and nonepidemic strains. Log reduction was negatively correlated with starting inoculum, indicating better survival of higher inocula. This study presents a framework to quantify bacterial survival. MRSA strains showed great capacity to persist in the environment, irrespective of epidemiological success. This finding strengthens the need for effective surface cleaning to contain MRSA transmission. IMPORTANCE Methicillin-resistant Staphylococcus aureus (MRSA) is a major cause of infections globally. While some MRSA clusters have spread worldwide, others are not able to disseminate successfully beyond certain regions despite frequent introduction. Dehydration tolerance facilitates transmission in hospital environments through enhanced survival on surfaces and fomites, potentially explaining differences in transmission success between MRSA clusters. Unfortunately, the currently available techniques to determine dehydration tolerance of cluster-forming bacteria like S. aureus are labor-intensive and unreliable due to their dependence on quantitative culturing. In this study, bacterial survival was assessed in a newly developed assay using isothermal microcalorimetry. With this technique, the effect of drying can be determined without the disadvantages of quantitative culturing. In combination with a newly developed mathematical algorithm, we determined dehydration tolerance of a large number of MRSA strains in a systematic, unbiased, and robust manner.

ABSTRACT Cervical cancer is the third leading cause of female cancers globally, resulting in more than 300,000 deaths every year. The majority of all cervical cancers are caused by persistent infections with high-risk human papillomaviruses (hrHPV) that can progress to cancer via a series of premalignant lesions. Most women, however, clear this infection within a year, concomitant with disease regression. Both hrHPV clearance and disease regression have been associated with the composition of the cervicovaginal microenvironment, which is defined by the host immune system and the cervicovaginal microbiome (CVM). A healthy microbiome is generally characterized by a high abundance of Lactobacillus species, and a change in the composition may cause bacterial vaginosis (BV), which is associated with an increased susceptibility to persistent hrHPV infections and disease. In this review, the composition of the CVM is discussed, with emphasis on the possible causes that drive changes in the cervicovaginal microbiota in relation to hrHPV infections, disease progression, and disease regression. The literature search focused on the composition of the CVM and its correlation with hrHPV infections and neoplastic lesions as well as the current efforts to adjust the microbiome against adverse viral outcomes.

The opportunistic pathogen Aspergillus fumigatus is found on all continents and thrives in soil and agricultural environments. Its ability to readily adapt to novel environments and to produce billions of spores led to the spread of azole-resistant A. fumigatus across the globe, posing a threat to many immunocompromised patients, including critically ill patients with severe influenza or COVID-19. In our study, we sought to compare the adaptational response to azoles from A. fumigatus isolates that differ in azole susceptibility and genetic background. To gain more insight into how short-term adaptation to stressful azole compounds is managed through gene expression, we conducted an RNA-sequencing study on the response of A. fumigatus to itraconazole and the newest clinically approved azole, isavuconazole. We observed many similarities in ergosterol biosynthesis up-regulation across isolates, with the exception of the pan-azole-resistant isolate, which showed very little differential regulation in comparison to other isolates. Additionally, we found differential regulation of membrane efflux transporters, secondary metabolites, iron metabolism, and various stress response and cell signaling mechanisms.

A parasitological study carried out in May 2022 and March 2023 in the Nyando River of Lake Victoria Basin, Kenya, disclosed two parasitic lernaeid copepods: Lamproglena cleopatra Humes, 1957, from the gills of a cyprinid, the Ningu Labeo victorianus Boulenger, 1901, endemic to the Lake Victoria drainage system, and Lamproglena clariae Fryer, 1957, from a clariid, the North African catfish Clarias gariepinus (Burchell, 1822). The copepods were studied and supplementary taxonomic information was presented using scanning electron micrographs and genetic data. Scanning electron microscopy (SEM) provided information on the morphology of L. cleopatra’s antennae, oral region, thoracic legs (2–5), and furcal rami not previously reported. Analyses of the partial fragments of 18S and 28S rDNA and cytochrome c oxidase subunit 1 (cox1) of the two parasites showed them to be distinct from all other Lamproglena taxa retrieved from GenBank. This study presents new taxonomic information on morphology using SEM and provides the first ribosomal (18S and 28S rDNA) and mitochondrial (mtDNA) data for these two parasite species. The cox1 data provided are the first for all 38 nominal species of Lamproglena. Notably, the study also provides a new host record for L. cleopatra and extends the geographical information of this species to Kenya.

Brevibacillus massiliensis strain phR is an obligately aerobic microbe that was isolated from human feces. Here, we show that it readily takes up tungsten (W), a metal previously associated only with anaerobes. The W is incorporated into an oxidoreductase enzyme (BmWOR) that was purified from native biomass. BmWOR consists of a single 65 kDa subunit and contains a single W-pyranopterin cofactor and a single [4Fe-4S] cluster. It exhibited high aldehyde-oxidizing activity with very high affinities (apparent Km

The pomegranate kernel oil has gained global awareness due to the health benefits associated with its consumption; these benefits have been attributed to its unique fatty acid composition. For quality control of edible fats and oils, various analytical and calorimetric methods are often used, however, these methods are expensive, labor-intensive, and often require specialized sample preparation making them impractical on a commercial scale. Therefore, objective, rapid, accurate, and cost-effective methods are required. In this study, Fourier transformed near-infrared (FT-NIR) and mid-infrared (FT-MIR) spectroscopy as a fast non-destructive technique was investigated and compared to qualitatively and quantitatively predict the quality attributes of pomegranate kernel oil (cv. Wonderful, Acco, Herskawitz). For qualitative analysis, principal component analysis (PCA) and orthogonal partial least squares discriminant analysis (OPLS-DA) was applied. Based on OPLS-DA, FT-MIR spectroscopy resulted in 100% discrimination between oil samples extracted from different cultivars. For quantitative analysis, partial least squares regression was used for model development over the NIR region of 7,498–940 and 6,102–5,774 cm−1 and provided the best prediction statistics for total carotenoid content (R2, coefficient of determination; RMSEP, root mean square error of prediction; RPD, residual prediction deviation; R2 = 0.843, RMSEP = 0.019 g β-carotene/kg, RPD = 2.28). In the MIR region of 3,996–1,118 cm−1, models developed using FT-MIR spectroscopy gave the best prediction statistics for peroxide value (R2 = 0.919, RMSEP = 1.05 meq, RPD = 3.54) and refractive index (R2 = 0.912, RMSEP = 0.0002, RPD = 3.43). These results demonstrate the potential of infrared spectroscopy combined with chemometric analysis for rapid screening of pomegranate oil quality attributes.

Abstract Staphylococcus aureus biofilms are a major problem in modern healthcare due to their resistance to immune system defenses and antibiotic treatments. Certain analgesic agents are able to modulate S. aureus biofilm formation, but currently no evidence exists if paracetamol, often combined with antibiotic treatment, also has this effect. Therefore, we aimed to investigate if paracetamol can modulate S. aureus biofilm formation. Considering that certain regulatory pathways for biofilm formation and virulence factor production by S. aureus are linked, we further investigated the effect of paracetamol on immune modulator production. The in vitro biofilm mass of 21 S. aureus strains from 9 genetic backgrounds was measured in the presence of paracetamol. Based on biofilm mass quantity, we further investigated paracetamol-induced biofilm alterations using a bacterial viability assay combined with N-Acetylglucosamine staining. Isothermal microcalorimetry was used to monitor the effect of paracetamol on bacterial metabolism within biofilms and green fluorescent protein (GFP) promoter fusion technology for transcription of staphylococcal complement inhibitor (SCIN). Clinically relevant concentrations of paracetamol enhanced biofilm formation particularly among strains belonging to clonal complex 8 (CC8), but had minimal effect on S. aureus planktonic growth. The increase of biofilm mass can be attributed to the marked increase of N-Acetylglucosamine containing components of the extracellular matrix, presumably polysaccharide intercellular adhesion. Biofilms of RN6390A (CC8) showed a significant increase in the immune modulator SCIN transcription during co-incubation with low concentrations of paracetamol. Our data indicate that paracetamol can enhance biofilm formation. The clinical relevance needs to be further investigated.

Background Direct access to hospital radiology facilities by general practitioner (GP) cooperatives is known to decrease the number of emergency department referrals, but the effects on length of stay (LOS; time from patient arrival at GP cooperative till departure to home) and patient experiences are unclear. Objectives To provide insight into the LOS and experiences of trauma patients with an indication for radiology at GP cooperatives with and without access to radiology. Methods A multi-methods observational study in April 2014–October 2015 at six GP cooperatives in The Netherlands, covering three organisational models for access to radiology: no direct access, limited access and unlimited access. Patient experiences were measured with a questionnaire. Patient records were analysed for background characteristics, radiology outcomes, referral and LOS. Results In total 657 patients were included, 232 no direct access model, 307 limited access model and 118 unlimited access model. The mean LOS was 99 minutes, with a significant difference between GP cooperatives without access to radiology (121 minutes), with limited access (86 minutes), and with unlimited access (90 minutes). The differences were larger for patients without radiological abnormalities. On a ten-point scale, patients rated GP cooperatives with unlimited access to radiology higher (8.62) than those without access (8.36) or with limited access (8.39). Conclusion Access to radiology by GP cooperatives seems to reduce the length of stay and is slightly more appreciated by patients. GP cooperatives with unlimited access seem to provide the most efficient and best-valued care, contributing to more patient-centred care.

Abstract Background Low concentrations of morphine are required for safe dosing for intrathecal injections. Sometimes, manual dilution of morphine is performed to achieve these low concentrations, but risks dilution errors and bacterial contamination. The primary goal was to compare the concentrations of morphine and bupivacaine between four groups of syringes. The secondary goal was to investigate the difference in contamination rate between these groups. Methods Twenty-five experienced anesthesia providers were asked to prepare a mixture of bupivacaine 2.0 mg/ml and morphine 60 μg/ml using 3 different methods as clean and precise as possible. The fourth method used was the aspiration of ampoules prepared by the pharmacy. The concentrations of morphine and bupivacaine were measured by High-Pressure Liquid Chromatography (HPLC). The medication was cultured for bacterial contamination. Results Group 1 (median 60 μg/ml; 95% CI: 59–110 μg/ml) yielded 3 outliers above 180 μg/ml morphine concentration. Group 2 (76 μg/ml; 95% CI: 72–80 μg/ml) and 3 (69 μg/ml; 95% CI: 66–71 μg/ml) were consistently higher than the target concentration of 60 μg. The group “pharmacy” was precise and accurate (59 μg/ml; 95% CI: 59–59 μg/ml). Group 2 and “pharmacy” had one contaminated sample with a spore-forming aerobic gram-positive rod. Conclusion Manually diluted morphine is at risk for deviating concentrations, which could lead to increased side-effects. Medication produced by the hospital pharmacy was highly accurate. Furthermore, even when precautions are undertaken, contamination of the medication is a serious risk and appeared to be unrelated to the dilution process.