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Development of a competition assay to assess the <i>in vitro</i> fitness of dengue virus serotypes using an optimized serotype-specific qRT-PCR

by Anne-Fleur Griffon, Loeïza Rault, Clément Tanvet, Etienne Simon-Lorière, Myrielle Dupont-Rouzeyrol, Catherine Inizan

Background

Comparing the in vitro fitness of dengue virus (DENV) isolates is a pivotal approach to assess the contribution of DENV strains’ replicative fitness to epidemiological contexts, including serotype replacements. Competition assays are the gold standard to compare the in vitro replicative fitness of viral strains. Implementing competition assays between DENV serotypes requires an experimental setup and an appropriate read-out to quantify the viral progeny of strains belonging to different serotypes.

Methods

In the current study, we optimized an existing serotyping qRT-PCR by adapting primer/probe design and multiplexing the serotype-specific qRT-PCR reactions, allowing to accurately detect and quantify all four DENV serotypes. We next developed an in vitro competition assay to compare the replicative fitness of two DENV serotypes in the human hepatic cell line HuH7.

Findings

The qRT-PCR was specific, and had a limit of detection below 7.52, 1.19, 3.48 and 1.36 genome copies/µL, an efficiency of 1.993, 1.975, 1.902, 1.898 and a linearity (R²) of 0.99975, 0.99975, 0.99850, 0.99965 for DENV-1, −2, −3 and −4, respectively. Challenge of this multiplex serotype-specific qRT-PCR on mixes of viral supernatants containing known concentrations of strains from two serotypes evidenced an accurate quantification of the amount of genome copies of each serotype. Quantification of the viral progeny of each serotype in the inoculum and the supernatant of competition assays using the serotype-specific multiplex qRT-PCR unveiled an enrichment of the supernatant in DENV-1 genome copies, uncovering the enhanced replicative fitness of this DENV-1 isolate.

Conclusions

This optimized qRT-PCR combined with a relevant cellular model allowed to accurately quantify the viral progeny of two DENV strains belonging to two different serotypes in a competition assay, allowing to determine which strain had a replicative advantage. This reliable experimental setup is adaptable to the comparative study of the replicative fitness of any DENV serotypes.

Psychosocial adjustment in patients with diabetic retinopathy in northeastern Iran: a qualitative study based on the transactional stress theory

Por: Salehnezhad · A.-S. · Sedghi · F. · Mahdizadeh · M. · Jamali · J. · Shoeibi · N. · Vahedian-Shahroodi · M.
Objectives

This study aimed to explore the psychosocial adaptation processes and coping methods among patients with diabetic retinopathy (DR), emphasising the importance of understanding their lived experiences.

Design

A qualitative study using semi-structured interviews.

Setting

A tertiary eye hospital in northeastern Iran.

Participants

The participants comprised patients with DR, their families and healthcare professionals.

Methods

Qualitative data were analysed based on transactional stress theory using deductive thematic analysis to identify psychosocial adaptation concepts.

Results

65% of participating patients had DR for more than 5 years. Analysis of 49 patient and 14 healthcare professional interviews revealed 6 key themes encompassing 15 categories and 33 subcategories: primary appraisal (threat perception, motivational assessment, self-blame), secondary appraisal (perceived control over disease outcomes, perceived control over emotions, self-efficacy), coping efforts (problem management, emotional regulation), meaning-based coping (positive reassessment, reinforcement of religious beliefs, constructive approach), moderators (seeking information, all-encompassing assistance) and adaptation (emotional health, individual-social functioning).

Conclusions

The analysis revealed that deficits in disease awareness, financial and systemic barriers and emotional distress significantly impeded adaptive coping. Conversely, self-efficacy, social and organisational support, and meaning-based coping strategies were key facilitators. Our findings suggest that facilitating positive psychosocial adjustment in patients with DR may require interventions that address these specific barriers and leverage these facilitators, such as providing comprehensive disease knowledge and constructing robust support systems. This study highlights the potential value of a holistic care approach that integrates medical treatment, targeted patient education and psychological support to improve the overall quality of life for these patients.

The LoaD study: the association between different types of physical activity and the progression of knee osteoarthritis--a cohort study protocol

Por: van der Voort · J. W. · Voogt · L. · Huisken · S. · Oei · E. · van Oeveren · B. · van Meurs · J. · Pluim · B. M. · Bierma-Zeinstra · S. M. A. · van Middelkoop · M.
Background

The global prevalence of osteoarthritis (OA) is expected to rise as the population ages, leading to increased physical impairments that often reduce physical activity. Knee OA, in particular, is a leading cause of chronic pain and disability in people over 40. OA progression varies among individuals, with factors such as higher age and body mass index contributing to a more rapid progression. However, research suggests that engaging in physical activity such as hiking and running may help alleviate pain and slow OA progression. However, current literature on the relationship between physical activity and knee OA is largely retrospective, lacks comparative analyses across different physical activity types and rarely considers the intensity of physical activity. To address these gaps, the LoaD study was designed as a prospective cohort study aimed at examining how different physical activity types (ie, hiking, running, cycling and tennis) and their intensities relate to knee OA progression over 24 months in physically active individuals with early signs of the condition.

Methods and analysis

This prospective cohort study will include 300 participants (75 per physical activity group), who will be followed for 24 months. Eligible participants are aged 45–65, meet the National Institute for Health and Care Excellence (NICE) criteria for clinical knee OA and engage in a minimum weekly activity level for either running (60 min), cycling (120 min), playing tennis (60 min) or hiking (1 hike of 10 km). Baseline and follow-up assessments will include MRI, physical examination (eg, biometrics, lower extremity strength) and blood sampling for molecular biomarker assessment (eg, genetics, inflammatory biomarkers). Participants will receive monthly online questionnaires (eg, demographics, lifestyle, general health, history of knee injuries) and their physical activities will be tracked continuously by global positioning system data from wearable devices. The study’s primary outcomes focus on knee OA progression and symptom changes. These will be assessed through (1) structural progression of knee OA, measured using the MRI Osteoarthritis Knee Score (MOAKS) and (2) clinical symptom progression evaluated by (a) the Knee Injury and Osteoarthritis Outcome Score sport and recreation function subscale and (b) Visual Analogue Scale for pain during an activity nominated by the participant to be the most aggravating for knee pain in the past month (VASNA, 0–10). Differences between groups on the primary outcomes will be analysed using mixed-effects models to determine the impact of activity type and intensity on knee OA progression.

Ethics and dissemination

The study is approved by the Erasmus MC Medical Research and Ethics Committee (MREC) (MEC-2022-0420). The results of this study will be made publicly available through peer-reviewed international open-access scientific journals and national and international conferences. Participants will be informed about the results as soon as they are published.

Continuous non-invasive electrophysiological monitoring in high-risk pregnancies: study protocol of a cohort intervention random sampling study in a tertiary obstetrical care centre in the Netherlands (NIEM-O study)

Por: de Klerk · N. D. · Berben · P. B. Q. · de Vries · I. R. · Niemarkt · H. · Vullings · R. · van den Heuvel · E. R. · van der Ven · M. · Fransen · A. F. · Oei · S. G. · van Laar · J. O. E. H.
Introduction

Women with high-risk pregnancies (eg, pre-eclampsia, imminent preterm birth) are often hospitalised due to the need for foetal and maternal monitoring. They are monitored for 30–45 min up to three times a day with conventional cardiotocography (CTG). In the meantime, they reside at the hospital, but the foetal status is not monitored. Continuous foetal monitoring is currently not recommended using CTG, due to the potential temperature rise from consistent exposure to ultrasound waves. For safe 24/7 monitoring, newly developed devices using non-invasive electrophysiological cardiotocography (eCTG) instead of conventional CTG offer a promising alternative. Previous research into eCTG has shown favourable results in monitoring foetal heart rate throughout both pregnancy and labour. This study aims to investigate the effect of implementing continuous antepartum eCTG monitoring in hospitalised high-risk pregnancies on perinatal and maternal outcome.

Methods and analysis

In this single centre prospective cohort intervention random sampling study, eligible women will be included on the Obstetric High Care of Máxima MC Veldhoven, the Netherlands. In total, 511 pregnant women with a singleton pregnancy between 23+0 and 32+0 weeks of gestation requiring hospitalisation will be recruited. Eligible women will be prospectively included in the cohort receiving standard treatment: intermittent CTG monitoring. From these women, a random sample of the prospective cohort will be offered a new monitoring method: 24/7 eCTG monitoring. For the eCTG monitoring, a wireless abdominal electrode patch, the Nemo Foetal Monitoring System will be used. Additional data from a historical cohort (2014–2019) of 1400 women receiving standard treatment will be included. Perinatal and maternal outcome, along with satisfaction levels of both patient and caregivers, will be compared between groups.

Ethics and dissemination

The study is registered on 18 October 2022 to the Central Committee on Research Involving Human Subjects (NL82869.015.22) via https://www.toetsingonline.nl/to/ccmo_monitor.nsf/conceptabr?OpenForm and approved by the Medical Ethics Committee of Máxima MC (W22.070) on 7 November 2023. Results of the study will be disseminated in peer-reviewed scientific journals and conference presentations.

Trial registration number

NCT06151613.

Exploring Health Care Needs and HR‐QOL Among Women With Cancer: A Principal Component Analysis Before and 6 Months After Treatment Initiation

ABSTRACT

Aim

To examine the characteristics of the health care needs corresponding to the medical care process and HR-QOL of women with cancer.

Design

A descriptive design was adopted.

Methods

The study's participants were 122 women with cancer who completed a survey before and 6 months after treatment initiation. A principal component analysis (PCA) was conducted on a set of 12 health care satisfaction scores at each point. Correlations were examined between the resulting components and HR-QOL indicators, including subjective well-being, symptoms, symptom-related interference, anxiety and depression.

Results

Most participants reported high health care satisfaction in both phases. PCA indicated the presence of 3 distinct domains: satisfaction with health care, health care management and supportive care. In both phases, these domains accounted for about 60% of the variance, while the remaining 40% was unexplained. Only satisfaction with health care was correlated with HR-QOL at both phases, with particularly strong associations observed for subjective well-being and depression at 6 months. Before treatment initiation, the item of ‘nursing care and practice’ received the highest average score, but demonstrated a negative loading on the component of ‘satisfaction with health care management’. The component of ‘satisfaction with supportive care needs’ was retained at both phases.

Conclusion

Health care plays a pivotal role in maintaining patients' quality of life, while supportive care and the integration of nursing practice within health care management remain essential.

Implications for Patient Care

High satisfaction scores do not necessarily mean that all health care needs are met. Addressing unmet needs from the perspective of HR-QOL and ensuring continuous supportive care throughout the treatment process is imperative.

Patient Contribution

Data provided by women with cancer was used.

Tensive Adaption: A Grounded Theory on the Process of Nurses' Dealing With Workplace Violence

ABSTRACT

Aim

To explore the process of how nurses experience and deal with workplace violence based on nurses' perceptions and experiences in Iran.

Methods

An exploratory qualitative study was conducted using grounded theory approach. Participants included 17 nurses working in 4 hospitals in 2 urban areas in Iran with at least 1 year of clinical experience in emergency departments and intensive care units. Data were collected through in-depth, semi-structured individual interviews conducted between August 2024 and March 2025. The constant comparative analysis approach was used for data analysis. This research method was carried out in five stages: open coding to identify concepts; development of concepts in terms of their characteristics and dimensions; contextual analysis; integration of the process to data analysis; and final category integration.

Findings

The main concern of participants in dealing with workplace violence was a ‘multidimensional security threat’. ‘Perpetrator response to nursing care’ (at the individual level) and ‘organisational inefficiency’ (at the organisational level) provided the context for this threat. A general theme entitled ‘tensive adaptation’ was the core category in this research and included four main categories: ‘tolerant reactions’, ‘seeking help’, ‘passive reactions’ and ‘hostile reactions’. ‘Organisational damage’, ‘nurse damage’ and ‘patient damage’ were the outcomes.

Conclusion

The theoretical model of ‘tensive adaptation’ provides a new perspective on ‘what’ and ‘how’ nurses experience and manage workplace violence. Effective strategies for managing violence such as effective communication, empathy, providing appropriate care, anger management, self-care, effective teamwork and requesting support can be considered by nursing administrators and incorporated into training programmes for nurses and nursing students. The adverse consequences of nurses' exposure to workplace violence should receive greater attention, as the entire healthcare system is affected by this exposure.

Implications for the Profession and/or Patient Care

Workplace violence leads to physical and psychological problems, reduced job satisfaction, diminished performance, negative effects on personal and family life and decreased quality of patient care. The main concern of participants in dealing with workplace violence was ‘multidimensional security threat’. This concept includes three characteristics: ‘psychological health threats’, ‘physical health threats’ and ‘professional position threat’. ‘Perpetrator response to nursing care’ (individual level) and ‘organisational inefficiency’ (organisational level) served as contextual conditions that expose nurses to workplace violence. In ‘tensive adaptation’, nurses attempt to respond to workplace violence through strategies such as ‘tolerant reactions’, ‘seeking help’, ‘passive reactions’ and ‘hostile reactions’. ‘Organisational damage’, ‘nurse damage’ and ‘patient damage’ are the consequences of nurses' exposure to workplace violence.

Reporting Method

The Consolidated Criteria for Reporting Qualitative Research (COREQ) was used to report this study.

Patient or Public Contribution

This study did not include patient or public involvement in its design, conduct, or reporting.

Barriers to mental health services for children and adolescents with autism spectrum disorder in Brazil: protocol for a qualitative evidence synthesis and citizen panel (BARRIER-Free-BR Project)

Por: Lopes · L. P. N. · Boeira · L. d. S. · Figueiredo Modesto · A. C. · Ramos-Silva · A. · Menin · V. P. · Abe · F. C. · Lopes · L. C.
Introduction

The perspectives of stakeholders directly affected by mental health services for autism spectrum disorder (ASD) are essential for the quality of these services. However, it is crucial that these perspectives are informed by the best available evidence and adapted to the local context. This study aims to analyse barriers related to mental health services for children and adolescents with ASD from the perspective of families and caregivers, considering social, racial and gender aspects.

Methods

Three steps will be taken: stakeholder engagement through an online meeting to refine the research question and understand the magnitude of the problem; (b) qualitative evidence synthesis using five databases and grey literature to identify studies that have collected and analysed qualitative data on barriers to mental health services for children and adolescents with ASD in Brazil. Only studies conducted in Brazil that consider the perspectives of family members and caregivers will be included. (c) A citizen panel with families of children and adolescents with ASD will be used to discuss and validate the synthesis findings.

Ethics and dissemination

We will provide a set of evidence-informed and stakeholder-experienced barriers to mental health services for children with ASD in Brazil. This represents an effort to engage stakeholders in evidence descriptions to inform policy. We plan to disseminate the findings through various means, including peer-reviewed journal publications, presentations at national conferences, invited workshops and webinars, patient associations and academic social media platforms. The project was approved by the Ethics Committee for Research at the University of Sorocaba (approval number 78747224.7.0000.5500).

Trial registration number

Open Science Framework—10.17605/OSF.IO/DVAKG.

Non-invasive electrophysiological monitoring vs conventional monitoring during labour in a tertiary obstetric care centre in the Netherlands: study protocol of a cohort intervention random sampling study (NIEM-II study)

Introduction

Conventional cardiotocography (CTG) has been used extensively to monitor the fetal condition during labour. However, conventional non-invasive monitoring is limited by the difficulty of obtaining an adequate signal quality, particularly in the case of obese parturients. Furthermore, the rate of operative deliveries keeps rising despite the ability for conventional intrapartum monitoring. Electrophysiological monitoring is an alternative technique that has been developed over the past decades to improve signal quality. This non-invasive, transabdominal and wireless alternative measures fetal heart rate by fetal electrocardiography (NI-fECG), and uterine activity by electrohysterography (EHG). Both NI-fECG and EHG have been proven to be more accurate and reliable than conventional non-invasive methods and are less affected by maternal Body Mass Index. Nevertheless, it is still unknown whether electrophysiological intrapartum monitoring leads to better obstetric and neonatal outcomes. This study aims to investigate whether electrophysiological monitoring during labour affects the number of operative interventions compared with conventional monitoring during labour.

Methods and analysis

This is a single-centre cohort intervention random sampling study which will be performed in a tertiary obstetric care centre. In total, 3471 term pregnant women with a singleton fetus in cephalic position and indication for continuous fetal monitoring during labour will be included. Eligible women will be prospectively included in the cohort for conventional monitoring. From these women, 90.9% of women will be randomly sampled and will be offered electrophysiological monitoring. A historical cohort of an additional 2100 women who received conventional monitoring will be added to the conventional group. This historical cohort was collected between April 2019 and February 2023. The primary outcome will be the number of operative interventions during labour. Secondary outcome measures include maternal and neonatal outcomes, patient and healthcare professional perspectives and costs.

Ethics and dissemination

This study received approval from the Medical Ethics Committee of Máxima Medical Centre (W22.071) on 1 November 2023. All participants will provide informed consent prior to data collection. Results of the study will be disseminated in peer-reviewed scientific journals and conference presentations.

Trial registration number

NCT06135961.

Preoperative partial breast reirradiation and repeat breast-conserving surgery in patients with recurrent breast cancer: the prospective single-arm REPEAT trial - a study protocol

Por: Civil · Y. · Wurfbain · L. · Jonker · L. · van der Sangen · M. · Oei · A. · Duvivier · K. · Bijker · N. · Meijnen · P. · van Kesteren · Z. · Palacios · M. · Barbe · E. · Menke-van der Houven van Oordt · W. · Diepenhorst · G. · Thijssen · V. · Slotman · B. · Verhoeff · J. · Schipper · R.
Introduction

Over the past decades, interest in second breast-conserving therapy (BCT) has increased due to, among others, advances in radiotherapy techniques. Preoperative partial breast irradiation (PBI) is an experimental treatment for patients with low-risk primary breast cancer. This approach can downstage the tumour and may possibly reduce toxicity and improve cosmetic outcomes compared with postoperative radiotherapy. This study aims to evaluate the feasibility of single-dose preoperative PBI and second breast-conserving surgery (BCS) for patients with an ipsilateral recurrent breast event (IRBE) after previous BCT.

Methods and analysis

The REPEAT trial is a multicentre, prospective, single-arm trial investigating ablative single-dose preoperative PBI in patients with an IRBE. Eligible patients are ≥50 years, have a unifocal non-lobular invasive breast cancer ≤2 cm, Bloom-Richardson grade 1 or 2, oestrogen receptor-positive, human epidermal growth factor receptor 2-negative and clinically negative axillary lymph nodes. The study plans to enrol 25 patients. Radiotherapy planning will involve the use of CT and MRI in the treatment position. Single-dose PBI of 20 Gy to the tumour and 15 Gy to the surrounding 2 cm of breast tissue will be delivered using a conventional or MR-guided linear accelerator. Tumour response will be monitored preoperatively using MRI and liquid biopsies to identify biomarkers for evaluating radiosensitivity. BCS will be performed 3 (±one) weeks post PBI. The primary endpoint is the incidence of grade 2 or higher treatment-associated acute toxicity within 90 days. Secondary endpoints include the evaluation of acute (grade 1) and late toxicity, radiologic and pathologic response, mastectomy rate, patient-reported outcomes, cosmetic outcome, local, regional and distant recurrence rates, survival outcome and biomarkers in liquid biopsies and tumour tissue. Patients will be followed up to 5 years after PBI.

Ethics and dissemination

Ethical approval from the Medical Research Ethics Committee of the Amsterdam UMC has been obtained (NL85983.018.24). The results will be disseminated via peer-reviewed academic journal and presentation at conferences. In addition, summaries will be shared with the participating patients.

Trial registration number

The trial was registered prospectively on October 11th 2024 at clinicaltrials.gov (NCT06640881).

Genicular nerve radiofrequency ablation, phenol neurolysis or conservative medical management in patients with knee osteoarthritis: protocol for the RADIOPHENOL randomised controlled multicentre trial with three parallel groups

Por: Wit · P. R. d. · Beek · R. v. · Schokker · M. · Wensing · C. · Hollmann · M. W. · Kallewaard · J.-W. · Oei · G. · RADIOPHENOL collaborators · Collaborative group name · Kampen · Elzinga · Hendriks · de Heiden · Godfried · Haumann · Thiel · Coumou
Introduction

Guidelines for symptomatic knee osteoarthritis (OA) dictate the initiation of conservative treatment (physical therapy, analgesics and intra-articular injections with corticosteroids) as a first line defence. When conservative treatment fails, the golden standard is invasive joint replacement surgery, but for a substantial group of patients who do not respond to the current conservative treatment, this is not (yet) indicated. The RADIOPHENOL study investigates if denervation of knee sensory (genicular) nerves can serve the gap between conservative and invasive treatment for younger patients and for patients who cannot undergo joint replacement surgery due to comorbid health conditions.

Methods and analysis

The RADIOPHENOL study is a multicentre unblinded randomised controlled trial with three parallel arms (1:1:1). In total, 192 patients with knee OA Kellgren-Lawrence grades 2–4 but not eligible for joint replacement according to the orthopaedic surgeon due to young age, old age and/or comorbidity or technical reasons are eligible and will be randomised to three groups of 64 patients. Group A: traditional radiofrequency ablation, group B: chemical neurolysis with phenol, group C: conservative medical management. Primary outcome is the Oxford Knee Score at 6 months. Secondary outcomes include Western Ontario and McMaster Universities Osteoarthritis Index, knee pain by numeric rating scale, physical functionality, health-related quality of life, mental health, change in medication use, predictive value of a diagnostic block, procedure time, patient discomfort score during the intervention and adverse events.

Ethics and dissemination

The protocol (V.2.0, 15 May 2023), was approved by the Ethics Committee of Amsterdam UMC (NL83410.018.22 – METC2022.0890) on 31 July 2023. We aim to publish our results in international peer-reviewed journals.

Trial registration details

ClinicalTrials.gov NCT06094660, including the WHO Trial Registration data set items. Registered on 20 October 2023, first patient enrolled on 27 November 2023.

Actividades de las enfermeras en cuidados intensivos: una revisión integradora

Objetivo principal: identificar qué actividades guían el trabajo de la enfermera en una unidad de cuidados intensivos para adultos. Metodología: revisión integradora, realizada sobre la base de Literatura Latinoamericana y del Caribe en Ciencias de la Salud y Sistema de Análisis y Recu-peración de Literatura Médica en línea, entre enero y febrero de 2019. La búsqueda resultó en 15 producciones, que fueron analizadas a través del análisis de contenido temático. Resultados principales: a partir del análisis de los estudios, surgió la siguiente categoría temática: “Actividades que guían el trabajo de la enfermera en una unidad de cuidados intensivos para adultos”, destacando: observación y vigilancia constantes, manejo de instrumentos tecnológicos e interpretación de información de estos dispositivos, desempeño y comunicación entre un equipo multidisciplinario, realizando una evaluación y plan de atención al paciente. Conclusión principal: aunque algunas actividades pueden caracterizarse como técnicas, también existen aquellas que involucran la subjetividad de las enfermeras, dirigidas a una atención más individualizada y guiadas por el ejercicio de la autonomía.

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