Exhaustive identification of genome-wide binding events of transcriptional regulators.

Genome-wide binding assays aspire to map the complete binding pattern of gene regulators. Common practice relies on replication-duplicates or triplicates-and high stringency statistics to favor false negatives over false positives. Here we show that duplicates and triplicates of CUT&RUN are not sufficient to discover the entire activity of transcriptional regulators. We introduce ICEBERG (Increased Capture of Enrichment By Exhaustive Replicate aGgregation), a pipeline that harnesses large numbers of CUT&RUN replicates to discover the full set of binding events and chart the line between false positives and false negatives. We employed ICEBERG to map the full set of H3K4me3-marked regions, the targets of the co-factor ß-catenin, and those of the transcription factor TBX3, in human colorectal cancer cells. The ICEBERG datasets allow benchmarking of individual replicates, comparing the performance of peak calling and replication approaches, and expose the arbitrary nature of strategies to identify reproducible peaks. Instead of a static view of genomic targets, ICEBERG establishes a spectrum of detection probabilities across the genome for a given factor, underlying the intrinsic dynamicity of its mechanism of action, and permitting to distinguish frequent from rare regulation events. Finally, ICEBERG discovered instances, undetectable with other approaches, that underlie novel mechanisms of colorectal cancer progression.

A new CUT&RUN low volume-urea (LoV-U) protocol optimized for transcriptional co-factors uncovers Wnt/ß-catenin tissue-specific genomic targets.

Upon WNT/ß-catenin pathway activation, stabilized ß-catenin travels to the nucleus where it associates with the TCF/LEF transcription factors, constitutively bound to genomic Wnt-responsive elements (WREs), to activate target gene transcription. Discovering the binding profile of ß-catenin is therefore required to unambiguously assign direct targets of WNT signaling. Cleavage under targets and release using nuclease (CUT&RUN) has emerged as prime technique for mapping the binding profile of DNA-interacting proteins. Here, we present a modified version of CUT&RUN, named LoV-U (low volume and urea), that enables the robust and reproducible generation of ß-catenin binding profiles, uncovering direct WNT/ß-catenin target genes in human cells, as well as in cells isolated from developing mouse tissues. CUT&RUN-LoV-U outperforms original CUT&RUN when targeting co-factors that do not bind the DNA, can profile all classes of chromatin regulators and is well suited for simultaneous processing of several samples. We believe that the application of our protocol will allow the detection of the complex system of tissue-specific WNT/ß-catenin target genes, together with other non-DNA-binding transcriptional regulators that act downstream of ontogenetically fundamental signaling cascades.

Stress-sensitive dynamics of miRNAs and Elba1 in Drosophila embryogenesis.

Early-life stress can result in life-long effects that impact adult health and disease risk, but little is known about how such programming is established and maintained. Here, we show that such epigenetic memories can be initiated in the Drosophila embryo before the major wave of zygotic transcription, and higher-order chromatin structures are established. An early short heat shock results in elevated levels of maternal miRNA and reduced levels of a subgroup of zygotic genes in stage 5 embryos. Using a Dicer-1 mutant, we show that the stress-induced decrease in one of these genes, the insulator-binding factor Elba1, is dependent on functional miRNA biogenesis. Reduction in Elba1 correlates with the upregulation of early developmental genes and promotes a sustained weakening of heterochromatin in the adult fly as indicated by an increased expression of the PEV wm4h reporter. We propose that maternal miRNAs, retained in response to an early embryonic heat shock, shape the subsequent de novo heterochromatin establishment that occurs during early development via direct or indirect regulation of some of the earliest expressed genes, including Elba1.

Single-cell response to Wnt signaling activation reveals uncoupling of Wnt target gene expression.

Wnt signaling drives nuclear translocation of ß-catenin and its subsequent association with the DNA-bound TCF/LEF transcription factors, which dictate target gene specificity by recognizing Wnt responsive elements across the genome. ß-Catenin target genes are therefore thought to be collectively activated upon Wnt pathway stimulation. However, this appears in contrast with the non-overlapping patterns of Wnt target gene expression in several contexts, including early mammalian embryogenesis. Here we followed Wnt target gene expression in human embryonic stem cells after Wnt pathway stimulation at a single-cell resolution. Cells changed gene expression program over time consistent with three key developmental events: i) loss of pluripotency, ii) induction of Wnt target genes, and iii) mesoderm specification. Contrary to our expectation, not all cells displayed equal amplitude of Wnt target gene activation; rather, they distributed in a continuum from strong to weak responders when ranked based on the expression of the target AXIN2. Moreover, high AXIN2 did not always correspond to elevated expression of other Wnt targets, which were activated in different proportions in individual cells. The uncoupling of Wnt target gene expression was also identified in single cell transcriptomics profiling of other Wnt-responding cell types, including HEK293T, murine developing forelimbs, and human colorectal cancer. Our finding underlines the necessity to identify additional mechanisms that explain the heterogeneity of the Wnt/ß-catenin-mediated transcriptional outputs in single cells.

Using mHealth applications for self-care - An integrative review on perceptions among adults with type 1 diabetes.

BACKGROUND: Individually designed interventions delivered through mobile health applications (mHealth apps) may be able to effectively support diabetes self-care. Our aim was to review and synthesize available evidence in the literature regarding perception of adults with type 1 diabetes on the features of mHealth apps that help promote diabetes self-care, as well as facilitators and barriers to their use. An additional aim was to review literature on changes in patient reported outcome measures (PROMs) in the same population while using mHealth apps for diabetes self-care. METHODS: Quantitative and qualitative studies focusing on adults aged 18 years and over with type 1 diabetes in any context were included. A systematic literature search using selected databases was conducted. Data was synthesised using narrative synthesis. RESULTS: We found that features of mHealth apps designed to help promote and maintain diabetes self-care could be categorized into self-care data monitoring, app display, feedback & reminders, data entry, data sharing, and additional features. Factors affecting the use of mHealth apps reported in the literature were personal factors, app design or usability factors, privacy and safety factors, or socioeconomic factors. Quality of life and diabetes distress were the most commonly reported PROMs in the included studies. CONCLUSION: We are unable to reach a conclusive result due to the heterogeneity of the included studies as well as the limited number of studies reporting on these areas among adults with type 1 diabetes. We therefore recommend further large-scale studies looking into these areas that can ultimately improve mHealth app use in type 1 diabetes self-care. SYSTEMATIC REVIEW REGISTRATION: Prospero CRD42020157620 .

The Meaning of Critical Illness for People Suffering From COVID-19: When a Frightening Unreality Becomes Reality.

The aim of this study was to elucidate the meaning of critical illness for people with COVID-19. This study used a qualitative design. Thirteen people who were critically ill with COVID-19 during 2020 and admitted to a COVID-19 intensive care unit in northern Sweden participated in the study. Data collection was conducted as individual interviews with a narrative approach, and data were analyzed with phenomenological hermeneutic interpretation. The participants did not think they would get critically ill with this unexpected illness. They experienced terrible nightmares where their relatives had been killed, and they missed their relatives both in their dreams and in reality, as they had not been allowed to be with them due to the virus. Gratefulness was described for surviving. Participants described thoughts of not being able to imagine going through this again. They felt fear and loneliness, as a terrifying unreality had become a reality.

The Swedish version of the TeamSTEPPS® teamwork attitudes questionnaire (T-TAQ): A validation study.

BACKGROUND: Effective teamwork is essential for delivering safe health care. It is important to increase patient safety in healthcare by conducting interprofessional team training with both healthcare professionals and undergraduate students. Validated questionnaires that evaluate team training activities contribute to valuable knowledge regarding changes in attitudes toward teamwork. The aim of the study was to test the reliability and structural validity of the Swedish version of the TeamSTEPPS® Teamwork Attitudes Questionnaire (T-TAQ). METHODS: The study had a cross-sectional design. Four hospitals in three health care regions in Sweden participated in the study. In total, 458 healthcare professionals, response rate 39.4%, completed the questionnaire. The T-TAQ, which consists of 30 items and covers five dimensions (Team Structure, Leadership, Situation Monitoring, Mutual Support and Communication), was translated to Swedish. A paper version of the T-TAQ was distributed to healthcare professionals (physicians, registered nurses, midwives, nursing assistants and allied health professionals) from the hospitals. Reliability and validity were tested using Cronbach's alpha and confirmatory factor analysis. RESULTS: Cronbach's alpha was 0.70 for the total T-TAQ and ranged from 0.41 to 0.87 for the individual dimensions. The goodness-of-fit indexes in the confirmatory factor analysis (Model 2) revealed a normed chi-square of 2.96, a root mean square error of approximation of 0.068, a Tucker-Lewis index of 0.785 and a comparative fit index of 0.808. CONCLUSIONS: The Swedish version of the T-TAQ has some potential to measure healthcare professionals' general attitudes toward the core components of teamwork in hospital settings. Further validation studies of the Swedish version of the T-TAQ are required, with samples representing both healthcare professionals and students from various healthcare disciplines and educational levels.

Psychosocial Self-efficacy and its Association with Selected Potential Factors Among Adults with Type 1 Diabetes: A Cross-Sectional Survey Study.

INTRODUCTION: The management of type 1 diabetes, a non-preventable chronic disease, leads to a high physical and psychological burden on the individual. Digital health technology can improve a person's psychosocial self-efficacy and thereby contribute to improved diabetes self-care. The aim of this study was to explore associations between psychosocial self-efficacy and demographic-, disease specific-, well-being as well as digital health technology (DHT) related factors among adults with type 1 diabetes. METHODS: A primarily web-based cross sectional survey was conducted among adults with type 1 diabetes in Sweden (n = 301). Psychosocial self-efficacy was assessed using the Swedish version of the Diabetes Empowerment Scale, Swe-DES-23. The survey also contained questions related to demographic-, disease specific-, well-being as well as digital health technology related variables. RESULTS: Higher well-being scores and lower HbA1c levels were associated with higher psychosocial self-efficacy in multiple linear regression analysis. In multivariate analysis, gender, body mass index, well-being scores, and HbA1c levels showed association with psychosocial self-efficacy. None of the DHT factors were found associated with psychosocial self-efficacy. CONCLUSIONS: In this study, higher well-being score and lower self-reported HbA1c levels were associated with higher psychosocial self-efficacy in both univariate- and multivariate analysis and accounted for 30% of the variation in psychosocial self-efficacy in the regression model. Thus, measures to improve psychosocial self-efficacy in adults with type 1 diabetes may help maintain their psychological well-being and blood glucose control.

Close relatives' perspective of critical illness due to COVID-19: Keeping in touch at a distance.

AIM: To elucidate the meaning of being a close relative of a critically ill person cared for in intensive care during the initial phase of the COVID-19 pandemic. DESIGN: A narrative inquiry design following the COREQ guidelines. METHODS: Individual interviews with fifteen close relatives of patients critically ill with COVID-19 were analysed using phenomenological hermeneutics. RESULTS: The surreal existence of not being allowed to be near was emotionally difficult. While distancing due to restrictions was challenging, physicians' phone calls served as a connection to their relatives and brought a sense of security. Keeping notes helped them remember what happened and brought order to a chaotic situation. CONCLUSION: Close relatives feel secure when they receive regular information about their critically ill relative, not just when their condition worsens. They wish to be physically near to their critically ill person; when this is impossible, digital technology can provide support, but further accessibility developments are needed.

Association of serum glial fibrillary acidic protein with progression independent of relapse activity in multiple sclerosis.

OBJECTIVE: Insidious disability worsening is a common feature in relapsing-remitting multiple sclerosis (RRMS). Many patients experience progression independent of relapse activity (PIRA) despite being treated with high efficacy disease-modifying therapies. We prospectively investigated associations of body-fluid and imaging biomarkers with PIRA. METHODS: Patients with early RRMS (n = 104) were prospectively included and followed up for 60 months. All patients were newly diagnosed and previously untreated. PIRA was defined using a composite score including the expanded disability status scale, 9-hole peg test, timed 25 foot walk test, and the symbol digit modalities test. Eleven body fluid and imaging biomarkers were determined at baseline and levels of serum neurofilament light (sNfL) and glial fibrillary acidic protein (sGFAP) were also measured annually thereafter. Association of baseline biomarkers with PIRA was investigated in multivariable logistic regression models adjusting for clinical and demographic confounding factors. Longitudinal serum biomarker dynamics were investigated in mixed effects models. RESULTS: Only sGFAP was significantly higher in PIRA at baseline (median [IQR] 73.9 [60.9-110.1] vs. 60.3 [45.2-79.9], p = 0.01). A cut-off of sGFAP > 65 pg/mL resulted in a sensitivity of 68% and specificity of 61%, to detect patients at higher risk of PIRA. In a multivariable logistic regression, sGFAP > 65 pg/mL was associated with higher odds of developing PIRA (odds ratio 4.3, 95% CI 1.44-12.86, p = 0.009). Repeated measures of sGFAP levels showed that patients with PIRA during follow-up had higher levels of sGFAP along the whole follow-up compared to stable patients (p < 0.001). CONCLUSION: Determination of sGFAP at baseline and follow-up may be useful in capturing disability accrual independent of relapse activity in early RRMS.

Midkine is associated with neuroendocrine differentiation in castration-resistant prostate cancer.

BACKGROUND: Castration-resistant prostate cancer (CRPC) is an incurable disease and both androgen-deprivation therapy (ADT) and neuroendocrine differentiation (NED) are closely related to CRPC transition. More knowledge concerning neuroendocrine (NE)-transformed PC cells, the NED process and its association with CRPC, is needed. Expression of growth factor midkine (MDK) is correlated with poor clinical outcomes in various human cancers, including PC. In the present study, we have evaluated MDK expression and NED in two separate tumor groups: early and advanced PC. METHODS: Immunohistochemical analysis of MDK, the neuronal marker tubulin-beta III (TUBB3) and the NE-marker chromogranin A (CGA) in a human archival material consisting of hormone naive (HN)/stage T1b (n = 29) and CRPC (n = 24) tumors. Triple immunofluorescent imaging was performed on a selection of specimens. RESULTS: MDK, TUBB3, and CGA were upregulated in CRPC compared to HN tumors. MDK was highly associated to the expression of both CGA and TUBB3, and identified MDK-positive NE-like looking cells found to co-express CGA or, more commonly, CGA together with TUBB3. CGA and TUBB3 staining displayed a partial expression overlap, an overlap almost exclusively displaying also MDK expression. CONCLUSIONS: MDK upregulation in CRPC is associated with NED (shown by its relation to CGA and TUBB3). The results suggest that MDK represents an over-bridging marker between different populations of NE-like tumor cells, possibly as part of the NED process and associated CRPC transition, something that needs to be evaluated experimentally as does the applicability of MDK as a future target.

Teriflunomide Concentrations in Cerebrospinal Fluid and Plasma in Patients with Multiple Sclerosis: A Pharmacokinetic Study.

BACKGROUND: Teriflunomide is a disease modifying treatment (DMT) approved for relapsing-remitting multiple sclerosis (RRMS) in adults and children. It reduces lymphocyte proliferation by inhibiting the mitochondrial enzyme dihydroorotate dehydrogenase (DHODH) and thereby the pyrimidine synthesis. Although most DMTs in multiple sclerosis (MS) modulate or inhibit the immune system in the periphery, the efficacy may improve if the agent also targets immune activity within the central nervous system (CNS), acts as a neuro-protective and enhances neuro-regeneration. The objective of this study was to determine the passage of teriflunomide over the blood-cerebrospinal fluid barrier (BCSFB). METHODS: Plasma and cerebrospinal fluid (CSF) teriflunomide concentrations were determined at steady state in 12 patients with RRMS, treated with oral teriflunomide 14 mg once daily. Included patients were all clinically stable without relapse or disability worsening within 6 months prior from baseline and were on no other immune modulating or immunosuppressive drugs. RESULTS: The mean teriflunomide concentrations in plasma and CSF were 38775 (SEM ± 7256) ng/mL and 68 (SEM ± 15) ng/mL, respectively. The passage over the BCSFB was 0.17 % (SEM ± 0.01). While no correlation was found between the function of the BCSFB assessed with the albumin ratio and the CSF teriflunomide concentration, the CSF and plasma teriflunomide concentrations were highly correlated (rs = 0.90, < 0.0001). CONCLUSIONS: Further studies are warranted to determine if the obtained CSF teriflunomide concentration reflects that in the CNS and is able to influence inflammatory and degenerative processes within the CNS.

The CUT&RUN suspect list of problematic regions of the genome.

BACKGROUND: Cleavage Under Targets and Release Using Nuclease (CUT&RUN) is an increasingly popular technique to map genome-wide binding profiles of histone modifications, transcription factors, and co-factors. The ENCODE project and others have compiled blacklists for ChIP-seq which have been widely adopted: these lists contain regions of high and unstructured signal, regardless of cell type or protein target, indicating that these are false positives. While CUT&RUN obtains similar results to ChIP-seq, its biochemistry and subsequent data analyses are different. We found that this results in a CUT&RUN-specific set of undesired high-signal regions. RESULTS: We compile suspect lists based on CUT&RUN data for the human and mouse genomes, identifying regions consistently called as peaks in negative controls. Using published CUT&RUN data from our and other labs, we show that the CUT&RUN suspect regions can persist even when peak calling is performed with SEACR or MACS2 against a negative control and after ENCODE blacklist removal. Moreover, we experimentally validate the CUT&RUN suspect lists by performing reiterative negative control experiments in which no specific protein is targeted, showing that they capture more than 80% of the peaks identified. CONCLUSIONS: We propose that removing these problematic regions can substantially improve peak calling in CUT&RUN experiments, resulting in more reliable datasets.

The four domains of the person-centred practice framework from the perspective of critical care nurses in intensive care units during a pandemic.

OBJECTIVES: The aim was to describe the Person-Centred Practice Framework's four domains (prerequisites, care environment, person-centred processes, and person-centred outcomes) through the perspectives of critical care nurses working in intensive care units during the second year of the COVID-19 pandemic. Furthermore, the aim was to investigate the relationships between prerequisites, care environment, person-centred processes, and person-centred outcomes. DESIGN/METHODS: A cross-sectional study involving questionnaires. Prerequisites were measured using person-related conditions, the care environment by using the Person-Centred Climate Questionnaire-Staff version, the person-centred processes by using the Person-Centred Care Assessment Tool and person-centred outcomes were measured with one question about present health and well-being and by using Self-rated Exhaustion Disorder. Descriptive and analytic statistics were used. Data was collected from July 2021 to November 2021. SETTING: Critical care nurses (n = 217) working in 15 Swedish adult intensive care units. RESULTS: Participants' average length of experience in intensive care units was 14 years, and most participants experienced increased nursing care responsibilities. They perceived the climate as safe but had limitations in terms of its everydayness and community. Participants perceived the organisations both supported and hindered personalized care. Most participants experienced a variety of exhaustion symptoms, and their health had positive relationship with community. CONCLUSION: By showing how prerequisites, care environment, person-centred process influences critical care nurses' health and well-being, organisations might identify aspects in the work environment that require targeted interventions to reach healthy workplaces. IMPLICATIONS FOR CLINICAL PRACTICE: To preserve the health and well-being of critical care nurses and to flourish as humans in their professional roles, they need to interact with and form relationships with their colleagues, patients, and relatives. Organisations should have a person-centred approach for every individual in the workforce to harness each critical care nurses' knowledge and skills for individuals to growth in their roles.

Moral Distress, Health and Intention to Leave: Critical Care Nurses' Perceptions During COVID-19 Pandemic.

Introduction: Moral distress increases the risk that critical care nurses will lose the ability to provide quality nursing care. Aims: To describe person-related conditions and perceptions of moral distress, health and intention to leave among critical care nurses in intensive care units, and to examine the relationship between person-related conditions, moral distress, health and intention to leave. Method: Cross-sectional, with 220 critical care nurses in 15 Swedish ICUs, and data gathered via a self-reported questionnaire. Results: Highest moral distress scores were reported in futile care and poor teamwork and 21% reported entertaining an intention to leave. Self-reported health was lower than before the COVID-19 pandemic and 4.1% reported pronounced exhaustion disorder. Self-reported health, reduced capacity to tolerate demands under time pressure, emotional instability or irritability, physical weakness, or being more easily fatigued and with decreased well-being were factors that had a relationship with futile care. Sleeping problems and intention to leave had a relationship with poor teamwork. Conclusions: Different strategies are needed to reduce moral distress and the leadership is crucial for managing crises such as the COVID-19 pandemic.

A mobile application for early labour support -feasibility pilot study.

BACKGROUND: Use of mobile applications (apps) are increasing during pregnancy but few of these are evidence-based or evaluated in research. AIM: To examine the feasibility, including perceived usefulness and usability, and the preliminary effects of an app based on the Confident birth method. METHODS: A mixed-method approach, including 48 women, was used to evaluate acceptability, usability and to test study design and procedures. iPhone-users (n = 24) tested the app during pregnancy while the remaining (n = 24) formed a control group. Background characteristics and outcome measurements were collected from all women at baseline. Women in the app group received two follow-up phone calls from a midwife concerning usefulness and ease of use of the app. A follow-up questionnaire after birth were used to measure preliminary effects of the intervention as well as system usability of the app. RESULTS: Women using the app found the app exercises simple, understandable, and useful. System usability score showed a mean score of 85.3 indicating excellent system usability. Notes from phone calls resulted in four categories: positive feedback about the app, negative feedback about the app, partners involvement, and knowledge. Preliminary effects of labour experience showed no significant differences between the two groups, in terms of early labour or childbirth experience. CONCLUSION: The app tested in this feasibility study, was perceived as useful and appreciated by women. Areas for improvement of the app were identified. The result shows promise for further efficacy testing in a forthcoming randomised controlled trial.

Measuring moral distress in Swedish intensive care: Psychometric and descriptive results.

OBJECTIVES: To investigate the construct validity and psychometric properties of the Swedish version of the Moral Distress Scale-Revised and to describe moral distress in an intensive care context. RESEARCH METHODOLOGY/DESIGN: The Italian Moral Distress Scale-Revised was translated and semantically adjusted to the Swedish intensive care context. A web survey with 14 moral distress items, as well as three additional and eight background questions was answered by critical care nurses (N = 71) working in intensive care units during the second year of the coronavirus disease pandemic. Inferential and descriptive statistics were used to investigate the Italian four-factor model and to examine critical care nurses' moral distress. RESULTS: The result shows a factor model of four components differing from the previous model. Critical care nurses demonstrated significant differences in moral distress regarding priorities compared to before the pandemic, type of household; experience as critical care nurses and whether they had supervised students during the pandemic. CONCLUSION: The component structure might have originated from the specific situation critical care nurses perceived during the pandemic. The health care organisations' role in preventing and healing the effects of moral distress is important for managers to understand. IMPLICATIONS FOR CLINICAL PRACTICE: Moral distress is common in intensive care and it is necessary to use valid instrument when measuring it. A psychometrical investigation of the Swedish version of the Moral Distress Scale-Revised, adapted for intensive care shows need for further semantic and cultural adaptation. Perceived priorities during the pandemic, household type, supervising during the pandemic and working experience were related to critical care nurses' experience of moral distress and managers need to be aware of conditions that may trigger such a response.

The time-resolved genomic impact of Wnt/ß-catenin signaling.

Wnt signaling orchestrates gene expression via its effector, ß-catenin. However, it is unknown whether ß-catenin binds its target genomic regions simultaneously and how this impacts chromatin dynamics to modulate cell behavior. Using a combination of time-resolved CUT&RUN against ß-catenin, ATAC-seq, and perturbation assays in different cell types, we show that Wnt/ß-catenin physical targets are tissue-specific, ß-catenin "moves" on different loci over time, and its association to DNA accompanies changing chromatin accessibility landscapes that determine cell behavior. In particular, Wnt/ß-catenin progressively shapes the chromatin of human embryonic stem cells (hESCs) as they undergo mesodermal differentiation, a behavior that we define as "plastic." In HEK293T cells, on the other hand, Wnt/ß-catenin drives a transient chromatin opening, followed by re-establishment of the pre-stimulation state, a response that we define as "elastic." Future experiments shall assess whether other cell communication mechanisms, in addition to Wnt signaling, are ruled by time, cellular idiosyncrasies, and chromatin constraints. A record of this paper's transparent peer review process is included in the supplemental information.

Early Labour App: Developing a practice-based mobile health application for digital early labour support.

BACKGROUND: Pregnant women in early labour have felt excluded from professional care, and their partners have been restricted from being involved in the birthing process. Expectant parents must be better prepared to deal with fear and stress during early labour. There is a need for evidence-based information and digital applications that can empower couples during childbirth. OBJECTIVE: To develop and identify requirements for a practice-based mobile health (mHealth) application for Digital Early Labour Support. METHODS: This research started with creating an expert group composed of a multidisciplinary team capable of informing the app development process on evidence-based practices. In consultation with the expert group, the app was built using an agile development approach (i.e., Scrum) within a continuous software engineering setting (i.e., CI/CD, DevOps), also including user and security tests. RESULTS: During the development of the Early Labour App, two main types of challenges emerged: (1) user challenges, related to understanding the users' needs and experience with the app, and (2) team challenges, related to the software development team in particular, and the necessary skills for translating an early labour intervention into a digital solution. This study reaffirms the importance of midwife support via blended care and the opportunity of complementing it with an app. The Early Labour App was easy to use, the women needed little to no help, and the partner's preparation was facilitated. The combination of the app together with blended care opens up awareness, thoughts and feelings about the method and provides good preparation for the birth. CONCLUSION: We propose the creation of the Early Labour App, a mHealth app for early labour support. The preliminary tests conducted for the Early Labour App show that the app is mature, allowing it to be used in the project's Randomised Control Trial, which is already ongoing.

A five-year observational prospective mono-center study of the efficacy of alemtuzumab in a real-world cohort of patients with multiple sclerosis.

Background: Alemtuzumab (ALZ) is a pulsed immune reconstitution therapy for multiple sclerosis (MS). Objective: To assess basic characteristics, therapeutic effects, and prognostic biomarkers on clinical and imaging parameters of disease activity for relapsing-remitting MS (RRMS) patients selected for ALZ, in a real-world long-term setting. Methods: Fifty-one RRMS patients [female = 31; mean age 36 (standard deviation 7.1) years; median expanded disability status scale (EDSS) 2 (interquartile range (IQR) 1.5)] initiating ALZ treatment, were consecutively included. Patients were assessed at baseline and thereafter annually for 5 years with clinical measures, symbol digit modality test (SDMT), and magnetic resonance imaging (MRI). Concentrations of glial fibrillary acidic protein (GFAP), reflecting astrogliosis, and neurofilament light (NfL), reflecting axonal damage, were measured in cerebrospinal fluid (CSF) and serum samples collected at baseline and after 2 years in CSF, and annually in serum. Control subjects were symptomatic controls (SCs, n = 27), who were examined at baseline and after 5 years without evidence of neurological disease. Results: While the mean annualized relapse rate was significantly reduced from baseline at each year of follow-up, disability was essentially maintained at a median EDSS of 1.5 and IQR between 1.13 and 2.25. New MRI activity was recorded in 26 patients (53%) over 5 years. The proportion of patients who achieved no evidence of disease activity (NEDA-3), 6-months confirmed disability worsening (CDW), and 6-months confirmed disability improvement (CDI) at 5 years were 33, 31, and 31%, respectively. The SDMT score was reduced for patients (p < 0.001), but unchanged for SCs. ALZ treatment did not change GFAP levels, whereas there was a significant decrease for RRMS patients in median CSF and serum NfL levels at follow-up [CSF month 24: 456 pg./mL (IQR 285.4) (p = 0.05); serum month 24: 6.7 pg/mL (IQR 4.7) (p < 0.01); serum month 60: 7.2 pg/mL (IQR 4.7) (p < 0.01)], compared to baseline [CSF: 1014 pg/mL (IQR 2832.5); serum 8.6 pg/mL (IQR 17.4)]. Conclusion: In this real-world mono-center population, we observed a progression-free survival of 69%, cumulative NEDA-3 of 33%, and reduced NfL levels, over a five-year follow-up. This confirms ALZ as an effective pulsed immune reconstitution therapy that significantly reduces neuro axonal loss, and therefore has the potential to reduce long-term neurological disability. ALZ did not appear to affect astrogliosis.