Methodological Protocol Research Articles

Impact of the COVID-19 pandemic on lifestyle’s behaviours of school-aged children in Portugal

Abstract Background COVID-19 pandemic impacted children’s daily routines. Based on this, WHO Collaborating Centre for Nutrition and Childhood Obesity in collaboration with WHO Regional Office for Europe coordinated the study about the impact of COVID-19 on lifestyle’s behaviours of school-age children in Europe, where 17 Member States participated, including Portugal. Methods Cross-sectional study was conducted within the sixth round of COSI Portugal, following a WHO common methodological protocol, during 2021-2022 school year. A national representative sample of families with children 6-8 years old were included and asked to report on variables comparing two periods: pre and during the COVID-19 pandemic. Results 5139 families participated. 11.4% reported a worsening of their perceived wealth and the percentage of fathers and mothers working full- or part-time decreased (74.6% to 68.7% and 70.6% to 59.6%, respectively) during pandemic. Children’s consumption of sweets (19.4%), savoury snacks (13.8%), fruit (11.9%) and vegetables (7.1%) increased, and sugary soft drinks (13.8%) decreased during pandemic. Family behaviours with the highest increases during pandemic were “eating together as a family” (35.4%), “cooking meals together with your child” (33.6%), “buying food in large quantities” (33.6%) and “eating home-cooked meals” (29.1%). Approximately, one in four parents reported children’s physical activity decreased and almost one in two reported an increase in recreational screen time. Parents who perceived their child as overweight during pandemic doubled (f7.9% to 14.6%). Children’s well-being behaviours that worsened more during pandemic were “having fun with friends” (44.3%), “got on well at school” (27.9%), “feel full of energy” (23.8%) and “feel fit and well” (22.2%). Conclusions These observations underscore the need to “build back better” post-pandemic with a focus on preventing and controlling childhood obesity and its determinants. Key messages • Portuguese children experienced negative impacts of the pandemic in their lifestyle behaviours, particularly on their physical health and well-being, with a report of a poorer socioeconomic situation. • This study and its findings give us a first-of-its-kind view into the physical and mental outcomes of the pandemic on school-aged children with a focus on the long-term, high-risk disease of obesity

Systematic review: locating qualitative academic publications for reviewing tenants’ and landlords’ renting experiences and interaction in the Majority World

This methodological protocol describes the step-by-step process of identifying the relevant international academic literature to be reviewed within the project ‘The affective economies of emerging private renting markets: understanding tenants and landlords in postcommunist Romania” (AFFECTIVE-PRS). It presents: (1) the preliminary decisions taken related to the breadth of the review (choice of databases, type of research, type of reference, searching fields); (2) the operationalisation of keywords and Boolean strings; (3) the further calibration of the searching parameters through piloting; (4) the final retrieval of relevant references through systematic and manual searches; and (5) the geographical coverage of the retained literature. While the paper demonstrates the rigour of the methodological approach taken, it also opens up the space for other scholars to scrutinise, replicate or adjust this approach to their own work.

The gendered value chain of matooke banana and its implications for tissue culture adoption in Uganda

ABSTRACT Tissue culture banana is promoted as a form of micro-propagation that can aid farmers in managing pests and disease in Uganda, the country with the largest per capita consumption of banana in the world. But uptake amongst smallholder farmers remains low. This study recruited 71 farmers from five banana-growing districts in Uganda to assess how gender dynamics of matooke production impact the adoption and sustained use of tissue culture. We collaborated with farmers and co-produced a three-stage, participatory methodological protocol for mapping value chains. Our analysis reveals a gendered division of labour and limited access to resources that shape farmers’ ability to benefit from this technology. Women farmers are interested in tissue culture banana but face challenges related to demands for land preparation, accessing plantlets, and relevant knowledge, while men are more reluctant due to their fragility, high cost, and the additional labour demands needed to ensure the crop's survival. This study highlights the necessity of understanding the gendered division along a crop’s value chain to ensure the success of new breeding technologies.

Les ambiances au sein des equipements touristiques de la ville coloniale de Biskra (Est Algerien)

Conquered by the French army in 1844, Biskra was widely known as a notorious winter tourist destination during the colonial era. Characterized by a hot and dry climate, the ‘queen of the oases’ in the Ziban region was renowned for its numerous tourist facilities constructed using techniques imported from elsewhere, local materials, and apparent local stylistic influences. This distinguished architecture was notably mentioned in travel accounts and highlighted in tourist guides. Situated in the realm of heritage and architecture, this research aims to go beyond the knowledge of the built (material) heritage of hotels and thermal resorts in a southern Algerian city. Therefore, this article identifies and defines the atmospheres (intangible heritage) and architectural components, along with their origins, within the tourist facilities built in Biskra during the colonial period. Content analysis is the research technique applied to a corpus of textual sources dating from the colonial period itself. Thus, this work is conducted in accordance with a methodological protocol allocating the study of atmospheres, form, and construction. The acquired results shed light on a historical mode of construction adapted to a challenging natural environment, and they provide a valuable historical reference for contemporary tourism projects suitable for demanding contexts.

Serious adverse events reporting in recent randomised clinical trials in intensive care medicine - A methodological study protocol.

Serious adverse events (SAEs) are common in intensive care unit (ICU) patients. Reporting of SAEs in randomised clinical trials (RCTs) varies why underreporting is likely. We aim to describe the reporting of SAEs from 2020 onwards and to illustrate the recent reporting of SAEs published in major medical journals. We will conduct a methodological study assessing pharmacological interventions in RCTs including adult ICU patients. We will search 10 general medical and critical care journals in PubMed. We will include all RCTs published from 2020 onwards. The primary research question is how many RCTs report SAEs in the primary publication. Secondary research questions include how SAEs are reported in the primary publication either as (1) proportion of patients experiencing one or more SAE, (2) all single events occurred, or (3) both strategies combined. We will assess the association between the proportion of patients with reported SAEs and the following trial characteristics: multicentred versus single-centre RCTs, industry-sponsored versus academic-sponsored, published trial protocol versus unpublished work, blinding, trials sample size, and RCTs focusing on COVID-19 patients versus other populations. The outlined methodological study will provide important information on the reporting of SAEs in recent drug trials in adult ICU patients.

Unveiling the Implications of Organic Nutrient Management Protocols on Soil Properties, Economic Sustainability, and Yield Optimization in Fenugreek Cultivation in Acidic Soils of Northeast India

Utilizing yield, profitability, and environmental sustainability in terms of soil quality as the goal variables, we created a methodological protocol for a thorough assessment of nutrient management (NM) technologies for feasibility and optimized fenugreek production employing organic sources of nutrients in the acid soil of Northeast India. Five organic nutrient management technologies were tried and tested comprising T1: absolute control; T2: 2.5 t ha−1 vermicompost (VC) + 250 kg ha−1 lime; T3: 5 t ha−1 farmyard manure (FYM) + 250 kg ha−1 lime; T4: 10 t ha−1 FYM + 250 kg ha−1 lime; and T5: 5 t ha−1 VC + 250 kg ha−1 lime with four replications laid out in a randomized block design for two consecutive growing seasons during 2018–2020. Results indicated that across the differential levels of organic amendments (treatments) employed, the addition of maximum doses of VC and FYM, in general, excelled over the other treatments concerning fenugreek seed yield, its yield attributes, soil properties, and better economic returns. Thus, the overall findings elucidated that the addition of higher doses of organic amendments (VC and FYM) can sustainably improve fenugreek productivity, soil properties, and economic returns for fenugreek growers in the acid soil of Northeast India.

Systematic review: locating qualitative academic publications for reviewing tenants' and landlords' renting experiences and interaction in the Majority World.

This methodological protocol describes the step-by-step process of identifying the relevant international academic literature to be reviewed within the project 'The affective economies of emerging private renting markets: understanding tenants and landlords in postcommunist Romania" (AFFECTIVE-PRS). It presents: (1) the preliminary decisions taken related to the breadth of the review (choice of databases, type of research, type of reference, searching fields); (2) the operationalisation of keywords and Boolean strings; (3) the further calibration of the searching parameters through piloting; (4) the final retrieval of relevant references through systematic and manual searches; and (5) the geographical coverage of the retained literature. While the paper demonstrates the rigour of the methodological approach taken, it also opens up the space for other scholars to scrutinise, replicate or adjust this approach to their own work.

Social acceptance in coastal and marine protected areas: Port-cros national park (France)

The effectiveness of marine protected areas is a timely issue, as the fight against biodiversity loss depends significantly on how effective these areas are. This article explores a key condition for meeting the challenge of improving conservation policies, as part of the global targets for biodiversity conservation, following the 2022 Kunming-Montreal Global Biodiversity Framework. The paper explores the determinants of social acceptance of Marine Protected Areas (MPAs) by local communities impacted directly by MPAs designation. We focus on the Port-Cros National Park (France), which was established in 1963. The park has seen its governance, perimeter and internal zoning evolve over time, generating local opposition and underlining the importance of acceptance by local communities. This article highlights the factors that play a role in the social acceptance of this National Park based on the results of a questionnaire survey including 569 residents of the municipalities in and around the protected area. The approach we followed is part of a European research project that has developed a methodological protocol to identify interactions between social parameters that characterise the level of support and has been applied in 20 national parks. Our results reveal a high level of social acceptance for the National Park, with four main factors influencing acceptance: perceptions of the social impacts of the park, trust in management authorities, sociodemographic characteristics and conditions for citizen participation. Spatial disparities are also demonstrated. Our analysis also enables us to explain the components of constrained acceptance: this opens possibilities to explore the conditions under which local communities can feel more empowered and participate in the governance of the protected area.

A systematic review and meta-analysis of cortisol levels in Plasmodium infections

Malaria has complex interactions with host physiology, including alterations in cortisol levels. Cortisol, a key hormone in the stress response, is known to be dysregulated in various infectious diseases. This systematic review and meta-analysis aimed to elucidate the relationship between Plasmodium infection and cortisol levels, shedding light on the intricate interplay between the parasite and the host’s endocrine system. The methodological protocol for assessing cortisol levels in malaria patients was registered in PROSPERO (CRD42024496578), a widely recognized international prospective register of systematic reviews. This registration ensures transparency and minimizes the risk of bias in our research. A comprehensive search strategy was employed across major databases, including Embase, PubMed, Scopus, and Medline, to include studies that reported cortisol levels in infected patients. The qualitative synthesis was undertaken to synthesize the difference in cortisol levels between malaria-infected and uninfected individuals. The meta-analysis employed the random effects model in the quantitative synthesis to calculate the effect estimate. The review included a total of 20 studies, with a substantial number conducted in Africa, followed by Asia and South America. Most included studies (13/20, 65%) reported higher cortisol levels in infected patients than in uninfected patients. The meta-analysis confirmed significantly higher cortisol levels in infected patients compared to uninfected individuals (P < 0.0001, standardized mean difference (SMD): 1.354, 95% confidence interval: 0.913 to 1.795, I2: 88.3%, across 15 studies). Notably, the method for cortisol measurement and the type of blood sample used (serum or plasma) were significant moderators in the analysis, indicating that these factors may influence the observed relationship between Plasmodium infection and cortisol levels. The systematic review and meta-analysis confirmed that Plasmodium infection is associated with increased cortisol levels, highlighting the intricate relationship between the disease and the host stress response. These findings underscore the potential of cortisol as a supplementary biomarker for understanding the pathophysiological impact of malaria. By providing insights into the stress-related mechanisms of malaria, this comprehensive understanding can inform future research and potentially enhance disease management and treatment strategies, particularly in regions heavily burdened by malaria.

A comprehensive protocol for efficient differentiation of human NPCs into electrically competent neurons

BackgroundThe study of neurons is fundamental to unraveling the complexities of the nervous system. Primary neuronal cultures from rodents have long been a cornerstone of experimental studies, yet limitations related to their non-human nature and ethical concerns have prompted the development of alternatives. In recent years, the derivation of neurons from human-induced pluripotent stem cells (hiPSCs) has emerged as a powerful option, offering a scalable source of cells for diverse applications. Neural progenitor cells (NPCs) derived from hiPSCs can be efficiently differentiated into functional neurons, providing a platform to study human neural physiology and pathology in vitro. However, challenges persist in achieving consistent and reproducible outcomes across experimental settings. Comparison with existing methodsOur aim is to provide a step-by-step methodological protocol, augmenting existing procedures with additional instructions and parameters, to guide researchers in achieving reproducible results. Methods and resultsWe outline procedures for the differentiation of hiPSC-derived NPCs into electrically competent neurons, encompassing initial cell density, morphology, maintenance, and differentiation. We also describe the analysis of specific markers for assessing neuronal phenotype, along with electrophysiological analysis to evaluate biophysical properties of neuronal excitability. Additionally, we conduct a comparative analysis of three different chemical methods—KCl, N-methyl-D-aspartate (NMDA), and bicuculline—to induce neuronal depolarization and assess their effects on the induction of both fast and slow post-translational, transcriptional, and post-transcriptional responses. ConclusionOur protocol provides clear instructions for generating reliable human neuronal cultures with defined electrophysiological properties to investigate neuronal differentiation and model diseases in vitro.

Experimental Designs in Management Studies for Bias Assessment: Setting and Stimulus Material

This manuscript stems from an ongoing research project on gender inequality in Portuguese businesses, particularly concerning access to leadership positions. It aims to provide new insights into the constraints that limit women’s access to managerial roles – both from the worker/employee’s perspective, by exploring expectations, aspirations, and perceived barriers, and from the employer/hierarchy’s perspective, by analysing which biases in recruitment and promotion processes are likely to limit women’s rise to top management positions. Against this analytical background, this paper aims to provide a reflection on the inquiry protocol developed within the project, with particular emphasis on the quantitative approach based on an experimental design. A survey has been developed targeting professionals and directors of large, listed companies and large/medium unlisted companies, where the respondents are given the framework of hiring hypothetical applicants for a management position. The methodological protocol involves elaborating different fictitious CV profiles, representing hypothetical candidates. These profiles are constructed by varying attributes according to various dimensions, specifically gender, combined with educational and professional characteristics. Based on the process of designing the experimental setting, the aim is to reflect on the nuances, obstacles, and limitations in constructing a set of stimulus materials and manipulating the experimental conditions regarding the fictitious applicants’ characteristics (including gender), vertical segregation (‘glass ceiling’ in access to top management positions), horizontal segregation (i.e., associating candidates with typically female or male occupations) and measurement items/scales for assessing the applicants in different dimensions (e.g., perceived competence, job fit, promotion recommendation, etc.). Therefore, this paper aims to reflect on the methodological challenges and advantages of an experimental research design and data analysis strategy to address gender bias in hiring contexts and capable of sustaining an ongoing data observatory to support the definition of public policies and organisational practices aimed at promoting gender equality in top positions.

The Identification and Documentation of On-site Sensory and Multisensory Experience–A Methodological Protocol

Single and multiple sense stimuli create sensescapes, which combine to be perceived as multisensory integrated products. Such encounters may be experienced across multiple spaces and have importance due to esthetic sensuality, cultural value, economic benefit, or religious significance. This article presents a methodological protocol for the identification and documentation of on-site sensory and multisensory experiences. It outlines currently accepted methodologies for the key senses, followed by an inventory of method benefits and limitations. Externalities affecting sensory surveys are discussed, and a decision tree is presented to plan for and execute a sensory survey.

Identification of Epigenetic Biomarkers of Adolescent Idiopathic Scoliosis Progression: A Workflow to Assess Local Gene Expression.

Adolescent idiopathic scoliosis (AIS) is a three-dimensional structural deformity of the spine that affects 2-3% of adolescents under the age of 16. AIS etiopathogenesis is not completely understood; however, the disease phenotype is correlated to multiple genetic loci and results from genetic-environmental interactions. One of the primary, still unresolved issues is the implementation of reliable diagnostic and prognostic markers. For clinical management improvement, predictors of curve progression are particularly needed. Recently, an epigenetic contribution to AIS development and progression was proposed; nevertheless, validation of data obtained in peripheral tissues and identification of the specific mechanisms and genes under epigenetic control remain limited. In this study, we propose a methodological approach for the identification of epigenetic markers of AIS progression through an original workflow based on the preliminary characterization of local expression of candidate genes in tissues directly involved in the pathology. The feasibility of the proposed methodological protocol has been originally tested here in terms of identification of the putative epigenetic markers of AIS progression, collection of the different tissues, retrieval of an appropriate amount and quality of RNA and DNA, and identification of suitable reference genes.

Linkage of routinely collected NHS data to evaluate liaison mental health services: challenges and lessons learned.

Liaison mental health services provide mental health care to patients in acute hospital settings. Evaluation of liaison services is challenging due to their heterogeneous organisation and delivery, high case throughput and varied patient case mix. We aimed to link routinely collected National Health Service data from secondary care settings, chosen for their service characteristics, to data from primary care to evaluate hospital-based liaison mental health services in England. We planned to compare patients referred to hospital-based liaison services with comparable patients in the same hospital not referred to liaison services and comparable patients in hospitals without any liaison services. We designed and enacted a methodology to link data from: (1) Hospital Episode Statistics, a database controlled by the National Health Service Digital and (2) ResearchOne, a primary care database controlled by The Phoenix Partnership. Obtaining approvals for the steps prespecified in the methodological protocol took 907 days. Enactment following approvals took 385 days. Data supplied from Hospital Episode Statistics contained 181,063 patients from 6 hospitals (mean=30,177, standard deviation=28,875.86) who matched the inclusion and exclusion criteria. Data supplied from ResearchOne contained 33,666 (18.6%) of these patients from the 6 hospitals (mean=5611, standard deviation=5206.59). Time required for approvals and enactment was attributable to slowness of data handling processes within each data holder and to resolution of technical and organisational queries between them. Variation in number of patients for which data was supplied between databases and between hospitals was attributable to coding inconsistencies and to the limited intersection of patient populations between databases and variation in recording practices between hospitals. Although it is technically feasible to link primary and secondary care data, the current system is challenging, complicated, unnecessarily bureaucratic, time consuming and costly. This limits the number of studies that could be conducted with these rich data sources. This article presents independent research funded by the National Institute for Health and Care Research (NIHR) Health and Social Care Delivery Research programme as award number 13/58/08.

The theory of social representations and the theory of mediations: a methodological proposal to draw both theories closer to each other

The article sumarizes research whose purpose was to bring together the theory of social representations, from Serge Moscovici, and the theory of mediations, from Jesús Martín-Barbero, with a view to studies in the field of Communication. A theoretical and methodological discussion is presented, based on a discussion of the main references of both theories. trying to point out points of tangency that allow the approximation of the theories. It results in a diagram as a methodological protocol suggestion for future studies.

REporting quality of PilOt randomised controlled trials in surgery (REPORTS): a methodological survey protocol

IntroductionThe aim of this methodological review is to evaluate the completeness of reporting of surgical pilot and feasibility randomised trials as per the Consolidated Standards of Reporting Trials (CONSORT) extension...

A methodological protocol for the development of a national guideline on perioperative management of gastrointestinal tumors in Germany.

The success of abdominal cancer surgery depends not only on the surgery itself but is influenced by the overall perioperative management. Given the multitude of perioperative measures and the ever-increasing number of studies on perioperative management, it is difficult to keep track and provide evidence-based perioperative management. The planned guideline on perioperative management will review the existing evidence and derive treatment recommendations. The processing of the evidence is carried out by 6 working groups according to an 8-step scheme: after drafting the guideline questions in PICO format (1), a systematic literature search is carried out (2), and the records found are screened by two independent reviewers from the coordination team. Subsequently, the full texts of the potentially relevant articles are made available to the working groups for full text screening (3). All articles to be included are reviewed for methodological quality (4) before summary of findings tables are generated (5). In line with the GRADE approach, confidence in the evidence is assessed (6) before a recommendation is derived from the evidence, using a modified GRADE Evidence to Decision Framework (7). Finally, all recommendations are compiled and agreed within the guideline group (8). Guidelines serve as foundation for therapy decisions in everyday clinical practice and should therefore be based on up-to-date research results. However, while primary studies and systematic reviews are critically reviewed for their methodological quality, the process of guideline development is often not comprehensible. A protocol with predefined methodology should therefore create transparency and strengthen confidence in the recommendations. The guideline is registered in the AWMF (Association of the Scientific Medical Societies) Guideline Register (088-010OL).

A step-by-step method to quantify coloration with digital photography

Coloration is often used in biological studies, for example when studying social signaling or antipredator defense. Yet, few detailed and standardized methods are available to measure coloration using digital photography. Here we provide a step-by-step guide to help researchers quantify coloration from digital images. We first identify the do's and don'ts of taking pictures for coloration analysis. We then describe how to i) extract reflectance values with the software ImageJ; ii) fit and apply linearization equations to reflectance values; iii) scale and select the areas of interest in ImageJ; iv) standardize pictures; and v) binarize and measure the proportion of different colors in an area of interest. We apply our methodological protocol to digital pictures of painted turtles (Chrysemys picta), but the approach could be easily adapted to any species. More specifically, we wished to calculate the proportion of red and yellow on the neck and head of turtles. With this protocol, our main aims are to make coloration analyses with digital photography:•More accessible to researchers without a background in photography.•More consistent between studies.

The Rat Thoracic Ultrasound protocol: scanning technique and normal findings.

Respiratory diseases (especially pneumonia) are very common disorders in pet rats. The suspected diagnosis is mostly based on the clinical signs, thoracic auscultation, and thoracic radiography. However, auscultation is insensitive in determining the severity of the disease, and radiographs are often unremarkable. Non-cardiac thoracic ultrasonography is increasingly used in veterinary medicine; however, it has not been described in detail in rats. Thoracic ultrasonic examination was conducted on 400 client-owned conscious pet rats. The rats were examined in the period from June 2023 to August 2023 in two veterinary clinics. Due to the small size of the animal, different anatomical considerations, and different evaluation protocols, as well as to meet the optimal outcome of detailed thoracic ultrasound, a standard methodological protocol was developed, and the name RATTUS (Rat Thoracic Ultrasound) was proposed. Typical signs of normal RATTUS were described (bat sign, lung sliding, A-lines, abdominal curtain sign, ski jump sign, lung pulse, seashore sign in M-mode, and bamboo sign). The new evaluation of lung inflation symmetry by substernal access was also described. The methodical approach presented and the normal findings description are proposed to be used for a standard/routine thoracic ultrasound examination in pet rats.

Qualitative methods and practices in transcultural research with forced migrants: the Asylum Seekers Photographic Interview (ASPI) methodological protocol

ABSTRACT The present study aims to describe the methodological procedures which led to the development of the ASPI, a photo-elicitation method aimed at exploring the life stories of Nigerian asylum seekers. The purposes of the manuscript are to share a research and methodological protocol for the development of similar methods as well as, through this, to extract and offer some methodological and operational research practices and criteria useful for transcultural research with forced migrants. The questioning attitude of researcher, intended as a methodological praxis in transcultural research, combined with the psychological function played by participatory research, which also engages a ‘third gaze’ on the traumatic experiences of asylum seekers, will be discussed to highlight and reflect upon the potentialities of qualitative research in this field.