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Abstract Heterochromatin organization is critical to many genome-related programs including transcriptional silencing and DNA repair. While super-resolution imaging, electron microscopy, and multiomics methods have provided indirect insights into the heterochromatin organization, a direct measurement of mesoscale heterochromatin ultrastructure is still missing. We use a combination of correlative light microscopy and cryo-soft X-ray tomography (CLXT) to analyze heterochromatin organization in the intact hydrated state of human mammary fibroblast cells. Our analysis reveals that the heterochromatin ultra-structure has a typical mean domain size of approximately 80 nm and a mean separation of approximately 120 nm between domains. Functional perturbations yield further insights into the molecular density and alterations in the mesoscale organization of the heterochromatin regions. Furthermore, our polymer simulations provide a mechanistic basis for the experimentally observed size and separation distributions of the mesoscale chromatin domains. Collectively, our results provide direct, label-free observation of heterochromatin organization in the intact hydrated state of cells.

Abstract Key message Genetic parameter estimates at age 12 years for trunk volume in Pinus pinaster displayed low to moderate heritability for growth and stem form, whereas wood properties showed high heritability. No correlation was detected between resistance to the nematode Bursaphelenchus xylophilus and growth, stem form, or wood properties. However, certain families stood out, showing favorable values across all traits, including resistance to the nematode. Context The breeding program for Pinus pinaster should advance towards a second-generation breeding population based on volume, stem form, and resistance to Bursaphelenchus xylophilus, with the challenge of incorporating novel traits of importance. Aims We focused on assessing the genetic parameters of growth, stem form, wood properties, and resistance to B. xylophilus as well as exploring the relationship between these traits. Methods We evaluated 116 half-sib families at three ages (7, 12, and 17 years) in seven progeny trials established in 2005, analyzing growth, stem form, and wood traits. Resistance to B. xylophilus was assessed in 91 of the 116 half-sib families at age 2 by artificial inoculation tests under greenhouse conditions. Results Advancing the selection age to 12 years proved to be feasible for volume selection in Galician P. pinaster. All traits, including resistance to B. xylophilus, showed potential for genetic improvement. Remarkably, resistance to B. xylophilus was not correlated to any other trait. Conclusion Excluding the most susceptible families to B. xylophilus from the Galician P. pinaster breeding program would not result in a significant decrease in volume, stem form, or wood properties.

Abstract Background Musculoskeletal alterations causing reduced range of motion of the ankle joint are common in children with cerebral palsy (CP). Objective measurements of passive joint resistance and three-dimensional skeletal muscle volume and muscle architecture can lead to a comprehensive understanding of which factors influence joint range of motion. Research question To investigate the relation between the passive dorsiflexion of the ankle joint, biomechanical contributing factors to the passive joint resistance, and muscular architectural properties of the triceps surae muscle in children with CP. Methods In this cross-sectional observational study, 14 children with spastic CP (bilateral: 5, unilateral: 9, Gross Motor Function Classification System (GMFCS) level I:11, II:3) naïve to intramuscular tone reducing treatment, and 14 TD children were included. The passive dorsiflexion of the ankle was measured with a goniometer. Passive joint resistance and related parameters were estimated based on a biomechanical model and measurements using a motorized device, the Neuroflexor. Three-dimensional muscle architecture was quantified with diffusion tensor magnetic resonance imaging (DT-MRI). Results In the CP group, the median [min, max] passive dorsiflexion was decreased in the most affected leg (MAL) compared to the less affected leg (LAL) (2.5° [-25°, 20°] vs. 12.5° [5°, 30°], p = 0.001). The stiffness coefficient (Nm/rad) in the MAL was significantly higher in children with CP compared to TD children (7.10 [3.39, 62.00] vs. 2.82 [1.24, 10.46], p = 0.015). Muscle architecture properties did not differ between CP and TD, except for pennation angle in the medial gastrocnemius (MG) of the MAL (CP 17.64° (2.29) vs. TD 21.46° (3.20), p = 0.017). The stiffness coefficient, in the MAL, correlated negatively to passive dorsiflexion (rs=-0.638) and pennation angle in medial gastrocnemius (rs=-0.964), and the non-linear coefficient (Non-linear 1) correlated negatively to the fascicle length of the medial gastrocnemius (rs=-0.857). Conclusion This study shows that stiffness of the plantarflexors is related to decreased passive dorsiflexion of the ankle and muscle structure of the MG in high-functioning children with spastic CP. Assessments of how dynamic components as well as microscopic muscle alterations contribute to joint stiffness in the plantarflexors in individuals with CP are warranted. Trial registration Retrospectively registered in ClinicalTrials.gov, NCT05447299. Observational study. Study start: 2019-01-15, register date: 2022-07-01.

Abstract Background Increasing nosocomial infections during the COVID-19 pandemic have been reported. However, data describing peripheral venous catheter associated bloodstream infections (PVC-BSI) are limited. Aims To describe the epidemiology and risk factors for PVC-BSI during the COVID-19 pandemic. Methods We conducted retrospective cohort study of prospectively collected PVC-BSI data in a 2100 bed hospital network in Switzerland. Adult patients with a PVC inserted between January 1, 2020 and December 31, 2021 were included. Risk factors for PVC-BSI were identified through descriptive analysis of patient and catheter characteristics, and univariable marginal Cox models. Results 206,804 PVCs and 37 PVC-BSI were analysed. Most PVC-BSI were attributed to catheters inserted in the Emergency department (76%) or surgical wards (22%). PVC-BSI increased in 2021 compared to 2020 (hazard ratio 2021 vs. 2020 = 2.73; 95% confidence interval 1.19–6.29), with a numerically higher rate of Staphylococcus aureus (1/10, 10%, vs. 5/27, 19%) and polymicrobial infection (0/10, 0% vs. 4/27, 15%). PVC insertions, patient characteristics, and catheter characteristics remained similar across the study period. PVC-BSI risk was associated with admission to the intensive care unit (ICU), and use of wide gauge catheter (

Abstract Background A large fraction of the disease burden in the Italian population is due to behavioral risk factors. The objective of this work is to provide a tool to estimate the impact of preventive interventions that reduce the exposure to smoking and sedentary lifestyle of the Italian population, with the goal of selecting optimal interventions. Methods We construct a Markovian model that simulates the state of each subject of the Italian population. The model predicts the distribution of subjects in each health status and risk factor status for every year of the simulation. Based on this distribution, the model provides a rich output summary, such as the number of incident and prevalent cases for each tracing disease and the Disability Adjusted Life Years (DALY), used to assess the impact of preventive interventions, and how this impact is shaped in time. Results This paper focuses on the methodological aspects of the model. The proposed model is flexible and can be applied to estimate the impact of complex interventions on the two risk factors and adapted to consider different cohorts. We validate the model by simulating the evolution of the Italian population from 2009 to 2017 and comparing the output with historical data. Furthermore, as a case-study, we simulate a counterfactual scenario where both tobacco and sedentary lifestyle are eradicated from the Italian population in 2019 and estimate the impact of such intervention over the following 20 years. Conclusions We propose a Markovian model to estimate how interventions on smoking and sedentary lifestyle can affect the reduction of the disease burden, and validate the model on historical data. The model is flexible and allows to extend the analysis to consider more risk factors in future research. However, we are aware that, given the ever-increasing availability of data, it is necessary in the future to increase the complexity of the model, to be closer to reality and to provide decision-making support to the policy-makers.

Abstract Endoscopic lung volume reduction (ELVR) using endobronchial valves (EBV) is a treatment option for a subset of patients with severe chronic obstructive pulmonary disease (COPD), suffering from emphysema and hyperinflation. In this pilot study, we aimed to determine the presence of bacterial biofilm infections on EBV and investigate their involvement in lack of clinical benefits, worsening symptomatology, and increased exacerbations that lead to the decision to remove EBVs. We analyzed ten COPD patients with ELVR who underwent EBV removal. Clinical data were compared to the microbiological findings from conventional EBV culture. In addition, EBV were analyzed by FISHseq, a combination of Fluorescence in situ hybridization (FISH) with PCR and sequencing, for visualization and identification of microorganisms and biofilms. All ten patients presented with clinical symptoms, including pneumonia and recurrent exacerbations. Microbiological cultures from EBV detected several microorganisms in all ten patients. FISHseq showed either mixed or monospecies colonization on the EBV, including oropharyngeal bacterial flora, Staphylococcus aureus, Pseudomonas aeruginosa, Streptococcus spp., and Fusobacterium sp. On 5/10 EBV, FISHseq visualized biofilms, on 1/10 microbial microcolonies, on 3/10 single microorganisms, and on 1/10 no microorganisms. The results of the study demonstrate the presence of biofilms on EBV for the first time and its potential involvement in increased exacerbations and clinical worsening in patients with ELVR. However, further prospective studies are needed to evaluate the clinical relevance of biofilm formation on EBV and appropriate treatment options to avoid infections in patients with ELVR.

Abstract Background The β-galactosidase from Paenibacillus wynnii (β-gal-Pw) is a promising candidate for lactose hydrolysis in milk and dairy products, as it has a higher affinity for the substrate lactose (low K M value) compared to industrially used β-galactosidases and is not inhibited by the hydrolysis-generated product D-galactose. However, β-gal-Pw must firstly be produced cost-effectively for any potential industrial application. Accordingly, the yeast Komagataella phaffii was chosen to investigate its feasibility to recombinantly produce β-gal-Pw since it is approved for the regulated production of food enzymes. The aim of this study was to find the most suitable way to produce the β-gal-Pw in K. phaffii either extracellularly or intracellularly. Results Firstly, 11 different signal peptides were tested for extracellular production of β-gal-Pw by K. phaffii under the control of the constitutive GAP promoter. None of the signal peptides resulted in a secretion of β-gal-Pw, indicating problems within the secretory pathway of this enzyme. Therefore, intracellular β-gal-Pw production was investigated using the GAP or methanol-inducible AOX1 promoter. A four-fold higher volumetric β-galactosidase activity of 7537 ± 66 µkat oNPGal/Lculture was achieved by the K. phaffii clone 27 using the AOX1 promoter in fed-batch bioreactor cultivations, compared to the clone 5 using the GAP promoter. However, a two-fold higher specific productivity of 3.14 ± 0.05 µkat oNPGal/gDCW/h was achieved when using the GAP promoter for β-gal-Pw production compared to the AOX1 promoter. After partial purification, a β-gal-Pw enzyme preparation with a total β-galactosidase activity of 3082 ± 98 µkat oNPGal was obtained from 1 L of recombinant K. phaffii culture (using AOX1 promoter). Conclusion This study showed that the β-gal-Pw was produced intracellularly by K. phaffii, but the secretion was not achieved with the signal peptides chosen. Nevertheless, a straightforward approach to improve the intracellular β-gal-Pw production with K. phaffii by using either the GAP or AOX1 promoter in bioreactor cultivations was demonstrated, offering insights into alternative production methods for this enzyme.

Abstract Mammalian DNA replication relies on various DNA helicase and nuclease activities to ensure accurate genetic duplication, but how different helicase and nuclease activities are properly directed remains unclear. Here, we identify the ubiquitin-specific protease, USP50, as a chromatin-associated protein required to promote ongoing replication, fork restart, telomere maintenance, cellular survival following hydroxyurea or pyridostatin treatment, and suppression of DNA breaks near GC-rich sequences. We find that USP50 supports proper WRN-FEN1 localisation at or near stalled replication forks. Nascent DNA in cells lacking USP50 shows increased association of the DNA2 nuclease and RECQL4 and RECQL5 helicases and replication defects in cells lacking USP50, or FEN1 are driven by these proteins. Consequently, suppression of DNA2 or RECQL4/5 improves USP50-depleted cell resistance to agents inducing replicative stress and restores telomere stability. These data define an unexpected regulatory protein that promotes the balance of helicase and nuclease use at ongoing and stalled replication forks.

Creutzfeldt–Jakob disease (CJD) is a rare, degenerative, and fatal brain disorder caused by abnormal proteins called prions. This research introduces a novel approach combining AI and graph theory to analyze histopathological microscopic images of brain tissues affected by CJD. The detection and quantification of spongiosis, characterized by the presence of vacuoles in the brain tissue, plays a crucial role in aiding the accurate diagnosis of CJD. The proposed methodology employs image processing techniques to identify these pathological features in high-resolution medical images. By developing an automatic pipeline for the detection of spongiosis, we aim to overcome some limitations of manual feature extraction. The results demonstrate that our method correctly identifies and characterize spongiosis and allows the extraction of features that will help to better understand the spongiosis patterns in different CJD patients.

Abstract Background Precision medicine has led to the development of targeted treatment strategies tailored to individual patients based on their characteristics and disease manifestations. Although precision medicine often focuses on a single health outcome for individualized treatment decision rules (ITRs), relying only on a single outcome rather than all available outcomes information leads to suboptimal data usage when developing optimal ITRs. Methods To address this limitation, we propose a Bayesian multivariate hierarchical model that leverages the wealth of correlated health outcomes collected in clinical trials. The approach jointly models mixed types of correlated outcomes, facilitating the “borrowing of information” across the multivariate outcomes, and results in a more accurate estimation of heterogeneous treatment effects compared to using single regression models for each outcome. We develop a treatment benefit index, which quantifies the relative benefit of the experimental treatment over the control treatment, based on the proposed multivariate outcome model. Results We demonstrate the strengths of the proposed approach through extensive simulations and an application to an international Coronavirus Disease 2019 (COVID-19) treatment trial. Simulation results indicate that the proposed method reduces the occurrence of erroneous treatment decisions compared to a single regression model for a single health outcome. Additionally, the sensitivity analyses demonstrate the robustness of the model across various study scenarios. Application of the method to the COVID-19 trial exhibits improvements in estimating the individual-level treatment efficacy (indicated by narrower credible intervals for odds ratios) and optimal ITRs. Conclusion The study jointly models mixed types of outcomes in the context of developing ITRs. By considering multiple health outcomes, the proposed approach can advance the development of more effective and reliable personalized treatment.

Abstract Current adeno-associated virus (AAV) gene therapy using nature-derived AAVs is limited by non-optimal tissue targeting. In the treatment of muscular diseases (MD), high doses are often required but can lead to severe adverse effects. Here, we rationally design an AAV capsid that specifically targets skeletal muscle to lower treatment doses. We computationally integrate binding motifs of human integrin alphaV beta6, a skeletal muscle receptor, into a liver-detargeting capsid. Designed AAVs show higher productivity and superior muscle transduction compared to their parent. One variant, LICA1, demonstrates comparable muscle transduction to other myotropic AAVs with reduced liver targeting. LICA1’s myotropic properties are observed across species, including non-human primate. Consequently, LICA1, but not AAV9, effectively delivers therapeutic transgenes and improved muscle functionality in two mouse MD models (male mice) at a low dose (5E12 vg/kg). These results underline the potential of our design method for AAV engineering and LICA1 variant for MD gene therapy.

Introduction: severe obesity results in high cardiovascular risk, increasing morbidity and mortality. New and improved methods are needed to detect cardiovascular diseases rapidly in severe obesity. microRNAs (miRNAs) has shown promise as diagnostic tools. This study aimed to identify plasma miRNAs useful as biomarkers of cardiovascular risk (CVR) in people with severe obesity. Methods: The study included 66 people with severe obesity classified in groups with atheroma (n = 32) and free of plaques (n = 34). Plasma samples were collected 1 month before bariatric surgery and at 6 and 12 months of follow-up. Participants were screened for the levels of 188 miRNAs, and 24 promising candidates were individually validated by quantitative polymerase chain reaction. Results: After validation, 5 of the 24 miRNAs showed significant differences over time in both groups: miR-375 increased after bariatric surgery, whereas miR-144-5p, miR-20a-3p, miR-145-5p and miR-21-3p exhibited decreased expression after bariatric surgery. The expression of 3 of the 24 miRNAs also differed between patients with and without atheroma: subjects with plaque had lower miR-126 but higher miR-21-3p and miR-133a-3p. Only miR-133a-3p exhibited exceptional discriminatory ability between subjects with and without plaque (area under the curve, 0.90; 95% confidence interval, 0.81–0.99). Conclusion: A specific signature of c-miRNA comprising miR-375, miR-144-5p, miR-20a-3p, miR-145-5p and miR-21-3p may facilitate CVR monitoring after bariatric surgery. Furthermore, miR-21-3p, miR-126-3p and miR-133a-3p show potential as specific biomarkers for subclinical atherosclerosis, with miR-133a-3p potentially able to diagnose subclinical atherosclerosis early in severe obesity.

ABSTRACTIn the face of the climate crisis, cities have committed to ambitious sustainability targets. The UN Agenda 2030 and its 17 Sustainable Development Goals (SDGs) provide a globally shared language for decision-makers and policymakers regarding their sustainability objectives. Espoo, Finland’s second-largest city, has become a pioneer in implementing the Agenda, not least due to its recent nomination as ‘Europe’s most sustainable city’. This article investigates the use of the Agenda in strategic governance in Espoo with the aim to identify challenges and opportunities of SDG localisation. Although the Agenda has affected many aspects of policymaking in Espoo and ambitions have been high to shine as an SDG pioneer, a systematic integration of sustainability concerns into policymaking is not yet achieved. Moreover, there is a need to question the reliance of the Agenda framework on the ecological modernisation paradigm, which does not see economic growth and ecological sustainability at odds.

Mental health today includes much more than the treatment of psychiatric disorders. More and more interventions aim to bring mental health support closer to people and psychotherapeutic interventions to people with somatic conditions. Since the treatment of people with metabolic syndrome and diabetes mellitus type 2 also requires a change in lifestyle, mental health has a prominent role. This overview paper wants to offer a solution after recognizing the given patterns where psychotherapy certainly has a significant and irreplaceable role. Precisely because of this phenomenon, psychological eating patterns associated with diabetes mellitus and hence metabolic syndrome should be studied, and attempts should be made to uncover patterns in occurrence. The aim of this study is to review the literature and consider the connection among diabetes mellitus, metabolic syndrome, and psychological eating patterns such as emotional and compulsive eating, as well as through the lens of food addiction. Furthermore, we have attempted to uncover the role of psychiatry and psychotherapy in the treatment of diabetes mellitus and metabolic syndrome and delve into the complexity of recognizing these patterns and emphasize the importance of a multidisciplinary approach in the treatment of diabetes mellitus and metabolic syndrome.

The abandonment of inhabited places is a phenomenon widespread on a global scale that has spanned centuries. It has led to the birth of the so-called ‘ghost towns’. These lifeless sites dot the internal Mediterranean and European areas, testifying to the changeability of the human settlements. Through a vision that reverses the paradigm that epitomises the ‘ghost towns’, these places can be transformed from a problem into an opportunity for the development of the territories that host them. The main topic of this article is to present and update investigations performed on three abandoned settlements sited in Portugal, Italy, and Albania in view of their tourist exploitation, considering three different tailored strategies: underwater tourism, dark tourism, and heritage tourism. For each site, we analysed the site history, the abandonment causes, and the territorial-geological features, thus arguing for the possible valorisation and enjoyment approaches with special attention to digital technologies, which are highly underexploited in the sector. This study, which is part of an international research landscape still in the initial stages, falls within the BEGIN project (aBandonment vErsus reGeneratIoN), which aims to develop a multilevel methodological–operational protocol useful in regional, national, EU, and non-EU contexts for the dissemination of knowledge, conservation, regeneration, valorisation, management, and fruition of areas classified as a ‘ghost town’. The research findings can be useful for scholars, practitioners, and local entities entrusted to manage the abandoned towns. Furthermore, the methodological approach followed in this research can supply useful insights into aims to valorise and enjoy worldwide ‘ghost towns’.

Abstract Pulmonary arterial hypertension (PAH) is a life-threatening disease with limited survival. Herein, we propose the pharmacological inhibition of Gq proteins as a novel concept to counteract pulmonary vasoconstriction and proliferation/migration of pulmonary artery smooth muscle cells (PASMCs) in PAH. We demonstrate that the specific pan-Gq inhibitor FR900359 (FR) induced a strong vasorelaxation in large and small pulmonary arteries in mouse, pig, and human subjects ex vivo. Vasorelaxation by FR proved at least as potent as the currently used triple therapy. We also provide in vivo evidence that local pulmonary application of FR prevented right ventricular systolic pressure increase in healthy mice as well as in mice suffering from hypoxia (Hx)-induced pulmonary hypertension (PH). In addition, we demonstrate that chronic application of FR prevented and also reversed Sugen (Su)Hx-induced PH in mice. We also demonstrate that Gq inhibition reduces proliferation and migration of PASMCs in vitro. Thus, our work illustrates a dominant role of Gq proteins for pulmonary vasoconstriction as well as remodeling and proposes direct Gq inhibition as a powerful pharmacological strategy in PH.

The effect of feed push-up has already been proven in several studies, so this topic can be considered one of the main points that helps improve the health status of dairy cattle. This study aimed to determine how the frequency of feed push-ups influences the health status of udder (mastitis), somatic cell counts, and reproduction. The effect of feed push-up on mastitis, the somatic cell counts, and the conception of dairy cows was evaluated. The feed was pushed-up at a frequency of 2x, 3x, 4x, 5x, and 6x a day for one calendar month. The effect on the number of dairy cows treated with mastitis was insignificant (P ≥ 0.05). The lowest number of cows with mastitis was found when feed was pushed-up five times daily. The effect on the somatic cell counts was insignificantly, too (P ≥ 0.05). The worst milk quality was found in the experimental group, which had a frequency of push-up 5x/day. However, it has been shown that the frequency of food push-up positively affected the conception rate in dairy cows (P < 0.001).

Abstract Background There is a need to understand the relationship between COVID-19 Convalescent Plasma (CCP) anti-SARS-CoV-2 IgG levels and clinical outcomes to optimize CCP use. This study aims to evaluate the relationship between recipient baseline clinical status, clinical outcomes, and CCP antibody levels. Methods The study analyzed data from the COMPILE study, a meta-analysis of pooled individual patient data from 8 randomized clinical trials (RCTs) assessing the efficacy of CCP vs. control, in adults hospitalized for COVID-19 who were not receiving mechanical ventilation at randomization. SARS-CoV-2 IgG levels, referred to as ‘dose’ of CCP treatment, were retrospectively measured in donor sera or the administered CCP, semi-quantitatively using the VITROS Anti-SARS-CoV-2 IgG chemiluminescent immunoassay (Ortho-Clinical Diagnostics) with a signal-to-cutoff ratio (S/Co). The association between CCP dose and outcomes was investigated, treating dose as either continuous or categorized (higher vs. lower vs. control), stratified by recipient oxygen supplementation status at presentation. Results A total of 1714 participants were included in the study, 1138 control- and 576 CCP-treated patients for whom donor CCP anti-SARS-CoV2 antibody levels were available from the COMPILE study. For participants not receiving oxygen supplementation at baseline, higher-dose CCP (/control) was associated with a reduced risk of ventilation or death at day 14 (OR = 0.19, 95% CrI: [0.02, 1.70], posterior probability Pr(OR

Abstract Background In vitro embryo production is a highly demanded reproductive technology in horses, which requires the recovery (in vivo or post-mortem) and in vitro maturation (IVM) of oocytes. Oocytes subjected to IVM exhibit poor developmental competence compared to their in vivo counterparts, being this related to a suboptimal composition of commercial maturation media. The objective of this work was to study the effect of different concentrations of secretome obtained from equine preovulatory follicular fluid (FF) on cumulus-oocyte complexes (COCs) during IVM. COCs retrieved in vivo by ovum pick up (OPU) or post-mortem from a slaughterhouse (SLA) were subjected to IVM in the presence or absence of secretome (Control: 0 µg/ml, S20: 20 µg/ml or S40: 40 µg/ml). After IVM, the metabolome of the medium used for oocyte maturation prior (Pre-IVM) and after IVM (Post-IVM), COCs mRNA expression, and oocyte meiotic competence were analysed. Results IVM leads to lactic acid production and an acetic acid consumption in COCs obtained from OPU and SLA. However, glucose consumption after IVM was higher in COCs from OPU when S40 was added (Control Pre-IVM vs. S40 Post-IVM: 117.24 ± 7.72 vs. 82.69 ± 4.24; Mean µM ± SEM; p

Education is seen as a resource at a global level but is currently considered to be in crisis in many parts of the world. This constitutes a significant drawback in terms of humanity’s prosperity and well-being since education is the key not only to an educated workforce but also to humane, collaborative, and caring societies. Even within this dim landscape, there are certain educational systems that defy the odds and perform significantly higher than their otherwise comparable systems. This paper proposes using an unusual lens for educational policy comparative studies, that of positive deviance, to aid us in progressing towards a more stable educational state of affairs. Using a positive deviance methodology, which focuses on learning what is working well in systems that defy and overcome substantial challenges, this study investigates the patterns, attitudes, and actions of three selected cases: Massachusetts as a positive deviant in the US, Estonia as a positive deviant in Europe, and Castile-Leon as a positive deviant in Spain. The purpose is, by analysing educational policies, laws, and other related documents, to find commonalities that explain why these systems outperform others. The results of the comparative analysis pinpoint areas and strategies informative to those leading struggling educational systems, such as a strong commitment to equity and justice, placing teachers at the centre of reforms, using assessment as a tool for process monitoring and summative inquiry, and making preschool education accessible to all.

Abstract Background The gram-positive bacterium Bacillus subtilis is widely used for industrial enzyme production. Its ability to secrete a wide range of enzymes into the extracellular medium especially facilitates downstream processing since cell disruption is avoided. Although various heterologous enzymes have been successfully secreted with B. subtilis, the secretion of cytoplasmic enzymes with high molecular weight is challenging. Only a few studies report on the secretion of cytoplasmic enzymes with a molecular weight > 100 kDa. Results In this study, the cytoplasmic and 120 kDa β-galactosidase of Paenibacillus wynnii (β-gal-Pw) was expressed and secreted with B. subtilis SCK6. Different strategies were focused on to identify the best secretion conditions. Tailormade codon-optimization of the β-gal-Pw gene led to an increase in extracellular β-gal-Pw production. Consequently, the optimized gene was used to test four signal peptides and two promoters in different combinations. Differences in extracellular β-gal-Pw activity between the recombinant B. subtilis strains were observed with the successful secretion being highly dependent on the specific combination of promoter and signal peptide used. Interestingly, signal peptides of both the general secretory- and the twin-arginine translocation pathway mediated secretion. The highest extracellular activity of 55.2 ± 6 µkat/Lculture was reached when secretion was mediated by the PhoD signal peptide and expression was controlled by the PAprE promoter. Production of extracellular β-gal-Pw was further enhanced 1.4-fold in a bioreactor cultivation to 77.5 ± 10 µkat/Lculture with secretion efficiencies of more than 80%. Conclusion For the first time, the β-gal-Pw was efficiently secreted with B. subtilis SCK6, demonstrating the potential of this strain for secretory production of cytoplasmic, high molecular weight enzymes.

Abstract Background Canada’s immigrants and refugees have often settled in large Canadian cities, but this is changing with rising costs of living and rural settlement initiatives. However, little consideration is made regarding systemic changes needed to accommodate this distribution, particularly in healthcare in medium-sized cities or smaller communities. For most Canadians, primary care is an entry point into the healthcare system but immigrants and refugees face unique barriers to accessing care compared to the general Canadian population. This project aimed to better understand the barriers to accessing primary care among newcomers in Peterborough, Ontario from the perspective of newcomer service providers. Methodology Participants were recruited from community organizations identified by the local settlement agency, the New Canadians Centre, as having regular interactions with newcomer clients including clinics, not-for-profit organizations, and volunteer groups. Four focus groups were completed, each with three participants (n=12). A coding grid was deductively developed to guide thematic analysis by adapting Levesque et al.’s conceptual framework defining access to healthcare with five specific dimensions: approachability, acceptability, availability and accommodation, affordability, and appropriateness. Results Participants identified lack of awareness of the healthcare system, stigma, competing priorities, and direct costs as some of the barriers for newcomers. Participants highlighted barriers unique to Peterborough including proximity to services, social isolation, and a shortage of family physicians. The results also highlighted strengths in the community such as its maternal-child health programming. Conclusion The results provide a glimpse of the challenges to accessing primary care among newcomers in medium-sized communities and identify opportunities to prepare for changing settlement patterns.

Background Available empirical evidence on participant-level factors associated with dropout from psychotherapies for post-traumatic stress disorder (PTSD) is both limited and inconclusive. More comprehensive understanding of the various factors that contribute to study dropout from cognitive-behavioural therapy with a trauma focus (CBT-TF) is crucial for enhancing treatment outcomes.Objective Using an individual participant data meta-analysis (IPD-MA) design, we examined participant-level predictors of study dropout from CBT-TF interventions for PTSD.Methods A comprehensive systematic literature search was undertaken to identify randomised controlled trials comparing CBT-TF with waitlist control, treatment-as-usual or another therapy. Academic databases were screened from conception until 11 January 2021. Eligible interventions were required to be individual and in-person delivered. Participants were considered dropouts if they did not complete the post-treatment assessment.Findings The systematic literature search identified 81 eligible studies (n=3330). Data were pooled from 25 available CBT-TF studies comprising 823 participants. Overall, 221 (27%) of the 823 dropped out. Of 581 civilians, 133 (23%) dropped out, as did 75 (42%) of 178 military personnel/veterans. Bivariate and multivariate analyses indicated that military personnel/veterans (RR 2.37) had a significantly greater risk of dropout than civilians. Furthermore, the chance of dropping out significantly decreased with advancing age (continuous; RR 0.98).Conclusions These findings underscore the risk of premature termination from CBT-TF among younger adults and military veterans/personnel.Clinical implication Understanding predictors can inform the development of retention strategies tailored to at-risk subgroups, enhance engagement, improve adherence and yield better treatment outcomes.

Abstract Fog and low stratus clouds (FLS) form as a result of complex interactions of multiple factors in the atmosphere and at the land surface and impact both the anthropogenic and natural environments. Here, we analyze the role of synoptic conditions and aerosol loading on FLS occurrence and persistence in the Po valley in northern Italy. By applying k‐means clustering to reanalysis data, we find that FLS formation in the Po valley is either based on radiative processes or moisture advection from the Mediterranean sea. Satellite‐based data on FLS persistence shows longer persistence of radiatively formed FLS events, likely due to air mass stagnation and a temperature inversion. Ground‐based aerosol optical depth observations further reveal that FLS event duration is significantly higher under high aerosol loading. The results underline the combined effect of topography, moisture advection and aerosol loading on the FLS life cycle in the Po valley.

Out-of-hospital cardiac arrest is one of the major health challenges faced by developed countries. Donation after circulatory death is a process of retrieving organs from individuals whose death has been confirmed by circulatory or respiratory criteria. In 2018, 136 625 people were listed on the waitlist covering over 16 countries. Out of these 136 625 individuals, 7383 died whilst waiting that year. The aim of this study is to assess the potential for the uncontrolled donation after circulatory death among out-of-hospital cardiac arrest patients in Slovenia. This non-interventional study was conducted using the Slovenian out-of-hospital cardiac arrest registry dataset. The database measured Out-of-hospital cardiac arrest in Slovenia between September and November 2022. From the database we chose patients who would be identified as donors by the uncontrolled donation after circulatory death guidelines for patient selection. Using the selection criteria we have narrowed the used data set from 294 unique patient records to 19. There were no organ donors in the cohort. With extrapolation we calculated that in 2022 there could be 111 donors in Slovenia that would fit the uncontrolled donation after circulatory death criteria. This equates to 52.4 pmp/y. We conclude that uncontrolled donation after circulatory death program in Slovenia would positively impact patients. Although our study is limited by a small sample of out-of-hospital cardiac arrest patients and short duration of the Slovenian out-of-hospital cardiac arrest registry inclusion, the results offer a good foundation to further explore uncontrolled organ donation in Slovenia and similar countries.

Background: Ongoing opioid treatment can potentially modify symptoms of myocardial infarction (MI) and cause a lack of recognition and treatment delay. Objectives: The purpose of this study was to examine MI symptoms and the time to hospitalization for patients in ongoing opioid treatment compared to patients without ongoing opioid treatment. Methods: We evaluated calls to the Copenhagen Emergency Medical Services in Denmark from 2014 to 2018. Calls were included when followed by hospitalization and a diagnosis of MI. Symptoms of MI and the time from call to hospitalization in patients in ongoing opioid treatment initiated prior to the onset of MI were compared to a control group of MI patients without opioid treatment. Results: In total, 6,633 calls were included; 552 calls from patients in opioid treatment and 6,081 calls from controls. Patients in opioid treatment were older and had more comorbidities than controls. Chest pain was less prevalent in MI patients in opioid treatment compared to controls (adjOR: 0.70; 95% CI: 0.57-0.85). The median time from the call to hospitalization was longer in patients in opioid treatment than in controls (50 vs 47 minutes; P = 0.006). Conclusions: In calls to the Emergency Medical Services, opioid treatment initiated prior to the onset of MI was associated with less frequent chest pain in MI. Therefore, awareness of ongoing opioid treatment may improve telephone triage of patients with MI, as symptom presentation in opioid-treated patients may differ and potentially challenge and delay the emergency response.

With the advent of Big Data, traditional digital computing is struggling to cope with intricate tasks related to data classification or pattern recognition. To mitigate this limitation, software‐based neural networks are implemented, but they are run in conventional computers whose operation principle (with separate memory and data‐processing units) is highly inefficient compared to the human brain. Brain‐inspired in‐memory computing is achieved through a wide variety of methods, for example, artificial synapses, spiking neural networks, or reservoir computing. However, most of these methods use materials (e.g., memristor arrays, spintronics, phase change memories) operated with electric currents, resulting in significant Joule heating effect. Tuning magnetic properties by voltage‐driven ion motion (i.e., magnetoionics) has recently emerged as an alternative energy‐efficient approach to emulate functionalities of biological synapses: potentiation/depression, multilevel storage, or transitions from short‐term to long‐term plasticity. In this perspective, the use of magnetoionics in neuromorphic applications is critically reviewed, with emphasis on modulating synaptic weight through: 1) control of magnetization by voltage‐induced ion retrieval/insertion; and 2) control of magnetic stripe domains and skyrmions in gated magnetic thin films adjacent to solid‐state ionic supercapacitors. The potential prospects in this emerging research area together with a forward‐looking discussion on future opportunities are provided.

Ovarian lymphangioma, a rare pathologic finding, is an ovarian mass characterized by lymphatic tissue lined with endothelial cells. It is normally asymptomatic and may be found incidentally during abdominal surgery for other pathologies.This report describes a case of a 49-year-old woman presenting to her primary care physician for three months of abdominal bloating and irregular menses. Magnetic resonance imaging revealed a 31 × 23 × 20 cm uterine mass suspected to be the cause of her symptoms. Total abdominal hysterectomy and bilateral salpingectomy were performed. During surgery, the right ovary was flattened and densely adhered to the body of the uterus, necessitating right oophorectomy. Pathology of the right ovary revealed flattened endothelial cells lining cystic spaces, consistent with the diagnosis of ovarian lymphangioma. Taken together, this case and the literature suggest that ovarian lymphangioma should be considered in the differential of ovarian masses, and their management shared more widely to help encourage the development of standard practice guidelines. There are no clear guidelines for when, and how often, to monitor these lesions after resection. In this case, the patient was seen at two-week and six-week follow-up visits with no new symptoms. Given that some case reports describe malignant transformation, patients should be followed this closely in the post-surgical period, and the best cadence for follow-up should be determined to improve outcomes.

Background Identifying patients at risk of nontuberculous mycobacterial pulmonary disease (NTM-PD) is challenging. Delays in NTM-PD identification and management are associated with declining lung function and increased morbidity and mortality. Study design and methods European NTM-PD experts (n=12) participated in a three-round modified Delphi process to score symptoms and comorbidities potentially associated with NTM-PD as reasons to test for nontuberculous mycobacteria. Results Experts reached a consensus on the symptoms and comorbidities that should and should not prompt testing for nontuberculous mycobacteria. Requirements for testing were scored as high (mean ≥7), medium (mean ≥4–

The use of chiral medicines (possessing center(s) of asymmetric carbon) may cause adverse drug reactions (ADRs). The safety assurance of these medicines is critical. We aimed to evaluate registered and commonly used anti-infective chiral medicines circulating in the Tanzanian market to establish their safety profile to protect public health. A mixed prospective-retrospective cohort study was conducted to assess the safety profile of amoxicillin, amoxicillin-clavulanic acid and ceftriaxone injection. ADRs causality assessment was conducted by using World Health Organization (WHO)-Algorithm criteria. Data were collected from 7 tertiary hospitals: Muhimbili National Hospital (MNH), Kilimanjaro Christian Medical Centre (KCMC), Bugando Medical Centre (BMC), Ligula Referral-Regional Hospital (LRRH), Kitete Referral-Regional Hospital (KRRH), Dodoma Referral-Regional Hospital (DRRH), and Mbeya Zonal-Referral Hospital (MZRH). Data were supplemented by those recorded in the WHO-Vigiflow/VigiLyze database within the same monitoring period. Data were analyzed using STATA version-15. The results were considered statistically significant when P

BackgroundFibromyalgia (FM) is a complex rheumatic disorder characterized by chronic nociplastic pain and central sensitization. Psychopathological conditions can influence FM symptoms, which worsen their condition. However, not all patients with FM have psychopathological disorders, indicating a heterogeneous population.ObjectiveTo investigate the psychopathological profile and personality disorders in patients with FM and its relationship impact on this disease.MethodsAn observational and cross-sectional comparative study was conducted with a sample of 90 women, mean age 48.7 years (SD = 8.12), from Hospital del Mar, Barcelona. The Personality Assessment Inventory (PAI) and the Fibromyalgia Impact Questionnaire (FIQ) were used for assessment.ResultsFM patients predominantly exhibited psychopathological profiles resembling affective disorders (37.7%) and Cluster C personality disorders (58.8%). The severity of FM’s impact was related to affective disorder symptoms, hypervigilance, derealization, somatization, and Cluster B personality disorder (emotional instability). Different rheumatic symptoms correlated with specific psychopathological patterns. Increased somatic symptoms on the FIQ were related to an unstable and dependent personality, while heightened emotional symptoms on the FIQ were associated with avoidance, borderline traits, and passive-aggressive reactions.ConclusionRecognizing psychopathological aspects is crucial for managing FM. The PAI is a valuable tool for establishing its psychopathological multidimensional profile, which predominantly shows an affective spectrum conditions and comorbid Cluster C personality disorder, exacerbating the disease’s impact.

ObjectiveThe cytochrome P450 2D6 (CYP2D6) is an enzyme involved in the oxidative biotransformation of various widely used drugs, including paroxetine, a substrate and strong inhibitor of the enzyme. The aim is to report on a case of protracted intoxication with paroxetine after a single overdose in a genotype-predicted intermediate CYP2D6 metabolizer.ObservationA 49-year-old man was receiving chronic treatment for more than 6 years with paroxetine 60 mg/day for an indication of agoraphobia. The patient ingested fifty 20 mg tablets of paroxetine in a suicide attempt. The toxic plasma level, accompanied by delirium, persisted for approximately 1 month after the overdose. According to the genotype profile, the patient was evaluated as an intermediate metabolizer with reduced CYP2D6 enzyme activity.ConclusionAs a consequence of the suicide attempt with overdose and the chronic paroxetine treatment that preceded it, phenoconversion to a poor metabolizer with very low CYP2D6 enzyme activity is suggested as contributing to an extremely long intoxication accompanied by delirium. Prolonged monitoring over a standard 24 h of both physical symptoms and drug plasma levels, together with a genetic profile assessment and phenoconversion consideration, is recommended after a single overdose in patients chronically treated with paroxetine.

This paper sheds light on the manufacturing processes, techniques, and materials used in the splendid illuminations of the oldest surviving copy of De Balneis Puteolanis, preserved at the Angelica Library in Rome (Ms. 1474). The codex is one of the masterpieces of mid-13th-century Italian-Southern illumination, traditionally referred to as the commission of Manfredi, son of Frederick II. The findings reported in the article result from the interdisciplinary study conducted in 2021–2023 in the framework of “Codex 4D: journey in four dimensions into the manuscript”, a multidisciplinary project involving many competences and dealing with art-historical studies on manuscripts, diagnostic and conservative analyses, scientific dissemination, storytelling, and public engagement. The considerations we present aims at increasing the knowledge of book artefacts while respecting their extraordinary complexity; data from non-invasive diagnostic investigations (X-ray fluorescence, Vis-NIR reflectance and Raman spectroscopies, hyperspectral imaging, and multi-band imaging techniques as ultraviolet, reflectography, and thermography), carried out in situ with portable instruments on the book, have been integrated with observations resulting from the historical-artistic study, and the reading of some ancient treatises on the production and use of the pigments and dyes employed in illumination.

Children are not miniature adults. Resuscitating a child can be stressful for the staff. The education of healthcare professionals is crucial for fast, sure, and correct interventions. After each resuscitation, a debriefing session is recommended.

Otroci niso pomanjšani odrasli. Oživljanje otroka je za osebje posebej stresno. Za hitro, suvereno in pravilno ukrepanje zdravstvenih delavcev je ključnega pomena izobraževanje. Po vsakem oživljanju je priporočljiv razbremenilni pogovor.

Abstract Background T-cell acute lymphoblastic leukemia (T-ALL) represents a rare and clinically and genetically heterogeneous disease that constitutes 10–15% of newly diagnosed pediatric ALL cases. Despite improved outcomes of these children, the survival rate after relapse is extremely poor. Moreover, the survivors must also endure the acute and long-term effects of intensive therapy. Although recent studies have identified a number of recurrent genomic aberrations in pediatric T-ALL, none of the changes is known to have prognostic significance. The aim of our study was to analyze the cytogenomic changes and their various combinations in bone marrow cells of children with T-ALL and to correlate our findings with the clinical features of the subjects and their treatment responses. Results We performed a retrospective and prospective comprehensive cytogenomic analysis of consecutive cohort of 66 children (46 boys and 20 girls) with T-ALL treated according to BFM-based protocols and centrally investigated cytogenetics and immunophenotypes. Using combinations of cytogenomic methods (conventional cytogenetics, FISH, mFISH/mBAND, arrayCGH/SNP and MLPA), we identified chromosomal aberrations in vast majority of patients (91%). The most frequent findings involved the deletion of CDKN2A/CDKN2B genes (71%), T-cell receptor (TCR) loci translocations (27%), and TLX3 gene rearrangements (23%). All chromosomal changes occurred in various combinations and were rarely found as a single abnormality. Children with aberrations of TCR loci had a significantly better event free (p = 0.0034) and overall survival (p = 0.0074), all these patients are living in the first complete remission. None of the abnormalities was an independent predictor of an increased risk of relapse. Conclusions We identified a subgroup of patients with TCR aberrations (both TRA/TRD and TRB), who had an excellent prognosis in our cohort with 5-year EFS and OS of 100%, regardless of the presence of other abnormality or the translocation partner. Our data suggest that escalation of treatment intensity, which may be considered in subsets of T-ALL is not needed for nonHR (non-high risk) patients with TCR aberrations.

Abstract Staphylococcal enterotoxin A (SEA) is the most frequently reported in staphylococcal food poisoning (SFP) outbreaks. Aptamers are single-stranded nucleic acids that are seen as promising alternatives to antibodies in several areas, including diagnostics. In this work, systematic evolution of ligands by exponential enrichment (SELEX) was used to select DNA aptamers against SEA. The SELEX protocol employed magnetic beads as an immobilization matrix for the target molecule and real-time quantitative PCR (qPCR) for monitoring and optimizing sequence enrichment. After 10 selection cycles, the ssDNA pool with the highest affinity was sequenced by next generation sequencing (NGS). Approximately 3 million aptamer candidates were identified, and the most representative cluster sequences were selected for further characterization. The aptamer with the highest affinity showed an experimental dissociation constant (KD) of 13.36 ± 18.62 nM. Increased temperature negatively affected the affinity of the aptamer for the target. Application of the selected aptamers in a lateral flow assay demonstrated their functionality in detecting samples containing 100 ng SEA, the minimum amount capable of causing food poisoning. Overall, the applicability of DNA aptamers in SEA recognition was demonstrated and characterized under different conditions, paving the way for the development of diagnostic tools.

Eruptions of volcanoes and geysers share many fundamental similarities: for example, they are manifestations of Earth’s geothermal energy, involving the pressure-driven expulsion of fluids from the Earth’s interior. However, while volcanoes can produce spectacular lava bubbles that burst, water bubbles are rarely observed on the surface of geysers. It is still unclear why some of these low-viscosity geyser systems produce none, while others produce them regularly. There is no quantification of the size, speed, and height of these bubbles at geysers, which is the gap we fill here. Strokkur creates a water bulge in its surface pool (bulge stage). When the bulge bursts, water is ejected into the air (jet stage). The steam then continues to rise buoyantly and drift away (drift stage). Here we study the evolution of the three stages using records from video camera campaigns and a local seismic network. We find that larger bulges are associated with larger ascent velocities and cause larger jet heights. As energy is channeled into a high jet, small seismic ground motions are recorded. The bulge formation itself is barely visible seismically. Our work suggests that the 0.74±0.27 s-long bulge stage can be used as a first-order proxy for predicting eruption height. This study might also be relevant for understanding fluid dynamics in volcanic systems.

OBJECTIVES: The aim of this study was to examine the mediating effect of the psychosocial work environment on the association between precarious employment (PE) and increased risk of common mental disorders (CMD), substance use disorders and suicide attempts. METHODS: This longitudinal register-study was based on the working population of Sweden, aged 25–60 years in 2005 (N=2 552 589). Mediation analyses based on a decomposition of counterfactual effects were used to estimate the indirect effect of psychosocial risk factors (PRF) (mediators, measured in 2005) on the association between PE (exposure, measured in 2005) and the first diagnosis of CMD, substance use disorders, and suicide attempts occurring over 2006–2017. RESULTS: The decomposition of effects showed that the indirect effect of the PRF is practically null for the three outcomes considered, among both sexes. PE increased the odds of being diagnosed with CMD, substance use disorders, and suicide attempts, among both men and women. After adjusting for PE, low job control increased the odds of all three outcomes among both sexes, while high job demands decreased the odds of CMD among women. High job strain increased the odds of CMD and suicide attempts among men, while passive job increased the odds of all three outcomes among women. CONCLUSION: The results of this study did not provide evidence for the hypothesis that psychosocial risks could be the pathways linking precarious employment with workers` mental health. Future studies in different social contexts and labour markets are needed.

Abstract Aims The aims of this sub‐study of the SMARTEX trial were (1) to evaluate the effects of a 12‐week exercise training programme on serum levels of high sensitivity cardiac troponin I (hs‐cTnI) in patients with moderate chronic heart failure (CHF), in New York Heart Association class II‐III with reduced ejection fraction (HFrEF) and (2) to explore the associations with left ventricular remodelling, functional capacity and filling pressures measured with N‐terminal pro brain natriuretic peptide (NT‐proBNP). Methods and results In this sub‐study, 196 patients were randomly assigned to high intensity interval training (HIIT, n = 70), moderate continuous training (MCT, n = 59) or recommendation of regular exercise (RRE), (n = 67) for 12 weeks. To reveal potential difference between structured intervention and control, HIIT and MCT groups were merged and named supervised exercise training (SET) group. The RRE group constituted the control group (CG). To avoid contributing factors to myocardial injury, we also evaluated changes in patients without additional co‐morbidities (atrial fibrillation, hypertension, diabetes mellitus, and chronic obstructive pulmonary disease). The relationship between hs‐cTnI and left ventricular end‐diastolic diameter (LVEDD), VO2peak, and NT‐proBNP was analysed by linear mixed models. At 12 weeks, Hs‐cTnI levels were modestly but significantly reduced in the SET group from median 11.9 ng/L (interquartile ratio, IQR 7.1–21.8) to 11.5 ng/L (IQR 7.0–20.7), P = 0.030. There was no between‐group difference (SET vs. CG, P = 0.116). There was a numerical but not significant reduction in hs‐cTnI for the whole population (P = 0.067) after 12 weeks. For the sub‐group of patients without additional co‐morbidities, there was a significant between‐group difference: SET group (delta −1.2 ng/L, IQR −2.7 to 0.1) versus CG (delta −0.1 ng/L, IQR −0.4 to 0.7), P = 0.007. In the SET group, hs‐cTnI changed from 10.9 ng/L (IQR 6.0–22.7) to 9.2 ng/L (IQR 5.2–20.5) (P = 0.002), whereas there was no change in the CG (6.4 to 5.8 ng/L, P = 0.64). Changes in hs‐cTnI (all patients) were significantly associated with changes in; LVEDD, VO2peak, and NT‐proBNP, respectively. Conclusions In patients with stable HFrEF, 12 weeks of structured exercise intervention was associated with a modest, but significant reduction of hs‐cTnI. There was no significant difference between intervention group and control group. In the sub‐group of patients without additional co‐morbidities, this difference was highly significant. The alterations in hs‐cTnI were associated with reduction of LVEDD and natriuretic peptide concentrations as well as improved functional capacity.

Background: Aging populations are driving a shift in emphasis toward enhancing chronic disease care, reflected in Catalonia’s regional plan which prioritizes standardized nursing care plans in primary care settings. To achieve this, the ARES-AP program was established with a focus on harmonizing standards and supporting routine nursing clinical decision-making. This study evaluates nurses’ perceptions of ARES-AP’s standardized care plans for chronic diseases. Methods: A mixed-methods approach based on an ad hoc questionnaire (n = 141) and a focus group (n = 14) was used. Quantitative data were statistically analysed, setting significance at p < 0.05. Qualitative data were explored via content analysis. Results: ARES-AP training was assessed positively. The resources for motivational interviewing and care plans for the most prevalent chronic diseases were rated very positively. This study identified key factors influencing program implementation, including facilitators such as structured information and nursing autonomy, barriers such as resistance to change, motivators such as managerial support, and suggested improvements such as technological improvements and time management strategies. Conclusions: This study identifies areas for improvement in implementing standardized nursing care plans, including additional time, motivation, enhanced IT infrastructure, and collaboration among primary care professionals. It enhances understanding of these plans in primary care, especially in managing chronic diseases in aging populations. Further research should assess the program’s long-term impact on chronic patients. This study was not registered.

Abstract Background Mantle cell lymphoma (MCL) is a chronically relapsing malignancy with deregulated cell cycle progression. We analyzed efficacy, mode of action, and predictive markers of susceptibility to palbociclib, an approved CDK 4/6 inhibitor, and its combination with venetoclax, a BCL2 inhibitor. Methods A panel of nine MCL cell lines were used for in vitro experiments. Four patient derived xenografts (PDX) obtained from patients with chemotherapy and ibrutinib-refractory MCL were used for in vivo proof-of-concept studies. Changes of the mitochondrial membrane potential, energy-metabolic pathways, AKT activity, and pro-apoptotic priming of MCL cells were evaluated by JC-1 staining, Seahorse XF analyser, genetically encoded fluorescent AKT reporter, and BH3 profiling, respectively. MCL clones with gene knockout or transgenic (over)expression of CDKN2A, MYC, CDK4, and RB1 were used to estimate impact of these aberrations on sensitivity to palbociclib, and venetoclax. Results Co-targeting MCL cells with palbociclib and venetoclax induced cytotoxic synergy in vitro and in vivo. Molecular mechanisms responsible for the observed synthetic lethality comprised palbociclib-mediated downregulation of anti-apoptotic MCL1, increased levels of proapoptotic BIM bound on both BCL2, and BCL-XL and increased pro-apoptotic priming of MCL cells mediated by BCL2-independent mechanisms, predominantly palbociclib-triggered metabolic and mitochondrial stress. Loss of RB1 resulted in palbociclib resistance, while deletion of CDKN2A or overexpression of CDK4, and MYC genes did not change sensitivity to palbociclib. Conclusions Our data strongly support investigation of the chemotherapy-free palbociclib and venetoclax combination as an innovative treatment strategy for post-ibrutinib MCL patients without RB1 deletion.

Abstract Background Adverse childhood experiences (ACEs), in combination with adverse community environments, can result in traumatic stress reactions, increasing a person’s risk for chronic physical and mental health conditions. Family resilience refers to the ability of families to withstand and rebound from adversity; it involves coping with disruptions as well as positive growth in the face of sudden or challenging life events, trauma, or adversities. This study aimed to identify factors contributing to family and community resilience from the perspective of families who self-identified as having a history of adversity and being resilient during the COVID-19 pandemic. Methods This study used Photovoice, a visual participatory research method which asks participants to take photographs to illustrate their responses to a research question. Participants consisted of a maximum variation sample of families who demonstrated family level resilience in the context of the pair of ACEs during the COVID-19 pandemic. Family members were asked to collect approximately five images or videos that illustrated the facilitators and barriers to well-being for their family in their community. Semi-structured in-depth interviews were conducted using the SHOWeD framework to allow participants to share and elucidate the meaning of their photos. Using thematic analysis, two researchers then independently completed line-by-line coding of interview transcripts before collaborating to develop consensus regarding key themes and interpretations. Results Nine families were enrolled in the study. We identified five main themes that enhanced family resilience: (1) social support networks; (2) factors fostering children's development; (3) access and connection to nature; (4) having a space of one’s own; and (5) access to social services and community resources. Conclusions In the context of additional stresses related to the COVID-19 pandemic, resilient behaviours and strategies for families were identified. The creation or development of networks of intra- and inter-community bonds; the promotion of accessible parenting, housing, and other social services; and the conservation and expansion of natural environments may support resilience and health.

The emergence of SARS-CoV-2 in 2019 and its rapid spread throughout the world has caused the largest pandemic of our modern era. The zoonotic origin of this pathogen highlights the importance of the One Health concept and the need for a coordinated response to this kind of threats. Since its emergence, the virus has caused >7 million deaths worldwide. However, the animal source for human outbreaks remains unknown. The ability of the virus to jump between hosts is facilitated by the presence of the virus receptor, the highly conserved angiotensin-converting enzyme 2 (ACE2), found in various mammals. Positivity for SARS-CoV-2 has been reported in various species, including domestic animals and livestock, but their potential role in bridging viral transmission to humans is still unknown. Additionally, the virus has evolved over the pandemic, resulting in variants with different impacts on human health. Therefore, suitable animal models are crucial to evaluate the susceptibility of different mammalian species to this pathogen and the adaptability of different variants. In this work, we established a transgenic mouse model that expresses the feline ACE2 protein receptor (cACE2) under the human cytokeratin 18 (K18) gene promoter's control, enabling high expression in epithelial cells, which the virus targets. Using this model, we assessed the susceptibility, pathogenicity, and transmission of SARS-CoV-2 variants. Our results show that the sole expression of the cACE2 receptor in these mice makes them susceptible to SARS-CoV-2 variants from the initial pandemic wave but does not enhance susceptibility to omicron variants. Furthermore, we demonstrated efficient contact transmission of SARS-CoV-2 between transgenic mice that express either the feline or the human ACE2 receptor.

Summary: Background: Bacille Calmette-Guérin (BCG) vaccination has off-target (non-specific) effects that are associated with protection against unrelated infections and decreased all-cause mortality in infants. We aimed to determine whether BCG vaccination prevents febrile and respiratory infections in adults. Methods: This randomised controlled phase 3 trial was done in 36 healthcare centres in Australia, Brazil, the Netherlands, Spain, and the United Kingdom. Healthcare workers were randomised to receive BCG-Denmark (single 0.1 ml intradermal injection) or no BCG in a 1:1 ratio using a web-based procedure, stratified by stage, site, age, and presence of co-morbidity. The difference in occurrence of febrile or respiratory illness were measured over 12 months (prespecified secondary outcome) using the intention-to-treat (ITT) population. This trial is registered with ClinicalTrials.gov, NCT04327206. Findings: Between March 30, 2020, and April 1, 2021, 6828 healthcare workers were randomised to BCG-Denmark (n = 3417) or control (n = 3411; no intervention or placebo) groups. The 12-month adjusted estimated risk of ≥1 episode of febrile or respiratory illness was 66.8% in the BCG group (95% CI 65.3%–68.2%), compared with 63.4% in the control group (95% CI 61.8%–65.0%), a difference of +3.4 percentage points (95% CI +1.3% to +5.5%; p 0.002). The adjusted estimated risk of a severe episode (defined as being incapacitated for ≥3 consecutive days or hospitalised) was 19.4% in the BCG group (95% CI 18.0%–20.7%), compared with 18.8% in the control group (95% CI 17.4%–20.2%) a difference of +0.6 percentage points (95% CI −1.3% to +2.5%; p 0.6). Both groups had a similar number of episodes of illness, pneumonia, and hospitalisation. There were three deaths, all in the control group. There were no safety concerns following BCG vaccination. Interpretation: In contrast to the beneficial off-target effects reported following neonatal BCG in infants, a small increased risk of symptomatic febrile or respiratory illness was observed in the 12 months following BCG vaccination in adults. There was no evidence of a difference in the risk of severe disease. Funding: Bill & Melinda Gates Foundation, Minderoo Foundation, Sarah and Lachlan Murdoch, the Royal Children's Hospital Foundation, Health Services Union NSW, the Peter Sowerby Foundation, SA Health, the Insurance Advisernet Foundation, the NAB Foundation, the Calvert-Jones Foundation, the Modara Pines Charitable Foundation, the UHG Foundation Pty Ltd, Epworth Healthcare, the National Health and Medical Research Council, the Swiss National Science Foundation and individual donors.

Multitarget ligands (MTLs) have emerged as an interesting alternative for addressing complex multifactorial pathologies such as neurodegenerative diseases. However, a common challenge associated with these compounds is often their high molecular weight and low solubility, which becomes a hurdle when trying to permeate over the blood-brain barrier (BBB). In this study, we have designed two new MTLs that modulate three pharmacological targets simultaneously (tau, beta-amyloid and TAR DNA-binding protein 43). To enhance their brain penetration, we have formulated organic polymeric nanoparticles using poly(lactic-co-glycolic acid). The characterization of the formulations, evaluation of their permeability through an in vitro BBB model, and assessment of their activity on disease-representative cellular models, such as Alzheimer’s disease and amyotrophic lateral sclerosis, have been conducted. The results demonstrate the potential of the new MTLs and their nanoparticle encapsulation for the treatment of neurodegenerative diseases.

This article provides a concise insight into the thermoforming of airlaid CTMP pulp. First, the airlaid process was studied, showing that fiber fractionation and the retention of fines occurred in the forming head. Then, the effect of temperature and pressure during thermoforming was investigated. Harsher conditions, i.e. higher temperature and pressure, yielded greater densification of the substrate and higher tensile strength. The maximum strength was found at the highest settings tested, that is, 100 MPa and 200°C. The screening of thermoforming conditions was also compared to previously published results on wetforming. Next, the effect of softwood CTMP pulp was delineated, which on average showed the best mechanical properties at elevated freeness and high degrees of bleaching. At last, a comparison between dry forming and wetforming was made for one selected pulp quality. Here, the dryformed substrates were stiffer at low elongation, yet the wetformed substrates yielded a greater extensibility and higher tensile strength. In conclusion, dryformed pulp mostly relies on temperature and pressure for bond formation during thermoforming, which produces materials that are distinctly different from wetformed molded pulp.