RAD51-dependent recruitment of TERRA lncRNA to telomeres through R-loops.

Telomeres-repeated, noncoding nucleotide motifs and associated proteins that are found at the ends of eukaryotic chromosomes-mediate genome stability and determine cellular lifespan1. Telomeric-repeat-containing RNA (TERRA) is a class of long noncoding RNAs (lncRNAs) that are transcribed from chromosome ends2,3; these RNAs in turn regulate telomeric chromatin structure and telomere maintenance through the telomere-extending enzyme telomerase4-6 and homology-directed DNA repair7,8. The mechanisms by which TERRA is recruited to chromosome ends remain poorly defined. Here we develop a reporter system with which to dissect the underlying mechanisms, and show that the UUAGGG repeats of TERRA are both necessary and sufficient to target TERRA to chromosome ends. TERRA preferentially associates with short telomeres through the formation of telomeric DNA-RNA hybrid (R-loop) structures that can form in trans. Telomere association and R-loop formation trigger telomere fragility and are promoted by the recombinase RAD51 and its interacting partner BRCA2, but counteracted by the RNA-surveillance factors RNaseH1 and TRF1. RAD51 physically interacts with TERRA and catalyses R-loop formation with TERRA in vitro, suggesting a direct involvement of this DNA recombinase in the recruitment of TERRA by strand invasion. Together, our findings reveal a RAD51-dependent pathway that governs TERRA-mediated R-loop formation after transcription, providing a mechanism for the recruitment of lncRNAs to new loci in trans.

The THO complex counteracts TERRA R-loop-mediated telomere fragility in telomerase+ cells and telomeric recombination in ALT+ cells.

Telomeres are the nucleoprotein structures at the ends of linear chromosomes. Telomeres are transcribed into long non-coding Telomeric Repeat-Containing RNA (TERRA), whose functions rely on its ability to associate with telomeric chromatin. The conserved THO complex (THOC) was previously identified at human telomeres. It links transcription with RNA processing, decreasing the accumulation of co-transcriptional DNA:RNA hybrids throughout the genome. Here, we explore the role of THOC at human telomeres, as a regulator of TERRA localization to chromosome ends. We show that THOC counteracts TERRA association with telomeres via R-loops formed co-transcriptionally and also post-transcriptionally, in trans. We demonstrate that THOC binds nucleoplasmic TERRA, and that RNaseH1 loss, which increases telomeric R-loops, promotes THOC occupancy at telomeres. Additionally, we show that THOC counteracts lagging and mainly leading strand telomere fragility, suggesting that TERRA R-loops can interfere with replication fork progression. Finally, we observed that THOC suppresses telomeric sister-chromatid exchange and C-circle accumulation in ALT cancer cells, which maintain telomeres by recombination. Altogether, our findings reveal crucial roles of THOC in telomeric homeostasis through the co- and post-transcriptional regulation of TERRA R-loops.

Health and inclusive labour force participation.

The future of work is rapidly changing, with higher flexibility of the labour market and increasing informal employment in many countries worldwide. There is also an increased pressure to extend working careers until older age. We introduce the concept of working life expectancy as a useful metric, capturing the expected numer of years in paid employment across the working age individuals, in particular among different groups. We describe factors that determine working life expectancy. Macro-level factors focus on the socioeconomic and political context that influences labour force participation, primarily policies and legislation in specific countries. At the meso level, employment contracts and working conditions are important. The micro level shows that individual characteristics, such as education, gender, and age, influence working careers. There are three important groups with a disadvantaged position in the labour market-workers with chronic diseases, workers with impairing disabilities, and workers aged 50 years or more. Within each of these disadvantaged groups, macro-level, meso-level, and micro-level factors that influence entering and exiting paid employment are discussed. To assure that paid employment is available for everyone of working age and that work contributes to better health, specific challenges need to be addressed at the macro, meso, and micro levels. To reach inclusive labour force participation, national policies, company practices, and workplace improvements need to be aligned to ensure safe and healthy workplaces that contribute to the health and wellbeing of workers and their communities.

A Stratified Approach for Managing Patients With Low Back Pain in Primary Care (SPLIT Program): A Before-and-After Study.

PURPOSE: To determine the effects of stratified primary care for low back pain (SPLIT program) in decreasing back-related disability for patients with low back pain (LBP) in primary care. METHODS: We conducted a before-and-after study. We compared health-related outcomes for 2 sequential, independent cohorts of patients with LBP recruited at 7 primary care units in Portugal. The first prospective cohort study characterized usual care (UC) and collected data from February to September 2018. The second was performed when the SPLIT program was implemented and collected data from November 2018 to October 2021. Between cohorts, physical therapists were trained in the implementation of the SPLIT program, which used the STarT Back Screening Tool to categorize patients for matched treatment. We compared back-related disability (Roland-Morris Disability Questionnaire, 0-24 points), pain (Numeric Pain Rating Scale, 0-10 points), perceived effect of treatment (Global Perceived Effect Scale, -5 to +5 points), and health-related quality of life (EuroQoL 5 dimensions 3 levels index, 0-1 points). RESULTS: We enrolled a total of 447 patients: 115 in the UC cohort (mostly treated with pharmacologic treatment) and 332 in the SPLIT cohort (all referred for a physical therapy intervention program). Over the study period of 6 months, patients in the SPLIT program showed significantly greater improvements in back-related disability (ß, -2.94; 95% CI, -3.63 to -2.24; P ≤ .001), pain (ß, -0.88; 95% CI, -1.18 to -0.57; P ≤ .001), perceived effect of treatment (ß, 1.40; 95% CI, 0.97 to 1.82; P ≤ .001), and health-related quality of life (ß, 0.11; 95% CI, 0.08 to 0.14; P ≤ .001) compared with UC. CONCLUSIONS: Patients in the SPLIT program for LBP showed greater benefits regarding health-related outcomes than those receiving UC.

Extracellular vesicle-based delivery of silencing sequences for the treatment of Machado-Joseph disease/spinocerebellar ataxia type 3.

Machado-Joseph disease (MJD)/spinocerebellar ataxia type 3 (SCA3) is the most common autosomal dominantly inherited ataxia worldwide. It is caused by an over-repetition of the trinucleotide CAG within the ATXN3 gene, which confers toxic properties to ataxin-3 (ATXN3) species. RNA interference technology has shown promising therapeutic outcomes but still lacks a non-invasive delivery method to the brain. Extracellular vesicles (EVs) emerged as promising delivery vehicles due to their capacity to deliver small nucleic acids, such as microRNAs (miRNAs). miRNAs were found to be enriched into EVs due to specific signal motifs designated as ExoMotifs. In this study, we aimed at investigating whether ExoMotifs would promote the packaging of artificial miRNAs into EVs to be used as non-invasive therapeutic delivery vehicles to treat MJD/SCA3. We found that miRNA-based silencing sequences, associated with ExoMotif GGAG and ribonucleoprotein A2B1 (hnRNPA2B1), retained the capacity to silence mutant ATXN3 (mutATXN3) and were 3-fold enriched into EVs. Bioengineered EVs containing the neuronal targeting peptide RVG on the surface significantly decreased mutATXN3 mRNA in primary cerebellar neurons from MJD YAC 84.2 and in a novel dual-luciferase MJD mouse model upon daily intranasal administration. Altogether, these findings indicate that bioengineered EVs carrying miRNA-based silencing sequences are a promising delivery vehicle for brain therapy.

MyBack - effectiveness and implementation of a behavior change informed exercise programme to prevent low back pain recurrences: a hybrid effectiveness-implementation randomized controlled study protocol.

BACKGROUND: Low back pain (LBP) is a common health condition and the leading cause of years lived with disability worldwide. Most LBP episodes have a favourable prognosis, but recurrences within a year are common. Despite the individual and societal impact related to LBP recurrences, there is limited evidence on effective strategies for secondary prevention of LBP and successful implementation of intervention programmes in a real-world context. The aim of this study is to analyse the effectiveness of a tailored exercise and behavioural change programme (MyBack programme) in the secondary prevention of LBP; and evaluate acceptability, feasibility and determinants of implementation by the different stakeholders, as well as the implementation strategy of the MyBack programme in real context. METHODS: This protocol describes a hybrid type I, randomized controlled trial to evaluate the effectiveness and implementation of MyBack programme in the context of primary health care. The Behaviour Change Wheel framework and FITT-VP principles will inform the development of the behaviour change and exercise component of MyBack programme, respectively. Patients who have recently recovered from an episode of non-specific LBP will be randomly assigned to MyBack and usual care group or usual care group. The primary outcome will be the risk of LBP recurrence. The secondary outcomes will include disability, pain intensity, musculoskeletal health, and health-related quality of life. Participants will be followed monthly for 1 year. Costs data related to health care use and the MyBack programme will be also collected. Implementation outcomes will be assessed in parallel with the effectiveness study using qualitative methods (focus groups with participants and health providers) and quantitative data (study enrolment and participation data; participants adherence). DISCUSSION: To our knowledge, this is the first study assessing the effectiveness and implementation of a tailored exercise and behaviour change programme for prevention of LBP recurrences. Despite challenges related to hybrid design, it is expected that data on the effectiveness, cost-effectiveness, and implementation of the MyBack programme may contribute to improve health care in patients at risk of LBP recurrences, contributing to direct and indirect costs reduction for patients and the health system. TRIAL REGISTRATION NUMBER: NCT05841732.

Characterization of a quail suspension cell line for production of a fusogenic oncolytic virus.

The development of efficient processes for the production of oncolytic viruses (OV) plays a crucial role regarding the clinical success of virotherapy. Although many different OV platforms are currently under investigation, manufacturing of such viruses still mainly relies on static adherent cell cultures, which bear many challenges, particularly for fusogenic OVs. Availability of GMP-compliant continuous cell lines is limited, further complicating the development of commercially viable products. BHK21, AGE1. CR and HEK293 cells were previously identified as possible cell substrates for the recombinant vesicular stomatitis virus (rVSV)-based fusogenic OV, rVSV-NDV. Now, another promising cell substrate was identified, the CCX.E10 cell line, developed by Nuvonis Technologies. This suspension cell line is considered non-GMO as no foreign genes or viral sequences were used for its development. The CCX.E10 cells were thus thoroughly investigated as a potential candidate for OV production. Cell growth in the chemically defined medium in suspension resulted in concentrations up to 8.9 × 106 cells/mL with a doubling time of 26.6 h in batch mode. Cultivation and production of rVSV-NDV, was demonstrated successfully for various cultivation systems (ambr15, shake flask, stirred tank reactor, and orbitally shaken bioreactor) at vessel scales ranging from 15 mL to 10 L. High infectious virus titers of up to 4.2 × 108 TCID50 /mL were reached in orbitally shaken bioreactors and stirred tank reactors in batch mode, respectively. Our results suggest that CCX.E10 cells are a very promising option for industrial production of OVs, particularly for fusogenic VSV-based constructs.

Transcranial sonography depicts a larger substantia nigra echogenic area in renal transplant patients on calcineurin inhibitors than on rapamycin.

BACKGROUND: After kidney transplantation neurologic manifestations may develop, including Parkinson's disease (PD). An enlarged substantia nigra (SN) by transcranial sonography has been recognized as a marker of PD. METHODS: In renal transplant recipients (RTRs = 95) and controls (n = 20), measurement of mesencephalon, SN, third ventricle, spleen and carotid intima-media thickness (cIMT) and middle cerebral artery (MCA), kidney and spleen arteries Doppler resistive index (RI) were performed. RESULTS: RTRs had larger SN, third ventricle and cIMT and higher renal RI than controls. The SN was larger in the CNIs group than in controls and rapamycin group, while the third ventricle was similar between patients but larger than in controls. In RTRs, SN showed a direct linear correlation with spleen and the third ventricle with age, cIMT and RI of the MCA, kidney and spleen. In CNIs group the SN correlated positively with age and cIMT, while the third ventricle reproduced RTRs correlations. Rapamycin group showed a direct linear relationship between the third ventricle and age and RI of the MCA, kidney and spleen; SN showed no correlations. CONCLUSION: RTRs on CNIs present a larger SN area than on rapamycin, probably due to the antiproliferative effect of rapamycin. This finding might be relevant when interpreting TCS in RTRs.

Synthesis of low-density polyethylene derived carbon nanotubes for activation of persulfate and degradation of water organic micropollutants in continuous mode.

Plastic derived carbon nanotubes (CNTs) were tested as catalysts in persulfate activation for the first time. Four catalysts were prepared by wetness impregnation and co-precipitation (using Al2O3, Ni, Fe and/or Al) and implemented to grow CNTs by chemical vapour deposition (CVD) using low-density polyethylene (LDPE) as carbon feedstock. A catalyst screening was performed in batch mode and the best performing CNTs (CNT@Ni+Fe/Al2O3-cp) led to a high venlafaxine mass removal rate (3.17 mg g-1 h-1) in ultrapure water after 90 min (even with a mixture of micropollutants). Its degradation increased when the matrix was replaced by drinking water and negligibly affected in surface water. A composite polymeric membrane was then fabricated with CNT@Ni+Fe/Al2O3-cp and polyvinylidene fluoride (PVDF), a high venlafaxine mass removal rate in surface water being also observed in 24 h of continuous operation. Therefore, the results herein reported open a window of opportunity for the valorisation of plastic wastes in this catalytic application performed in continuous mode.

Cyanine Dye Coupling Mediates Self-assembly of a pH Sensitive Peptide into Novel 3D Architectures.

Synthetic multichromophore systems are of great importance in artificial light harvesting devices, organic optoelectronics, tumor imaging and therapy. Here, we introduce a promising strategy for the construction of self-assembled peptide templated dye stacks based on coupling of a de novo designed pH sensitive peptide with a cyanine dye Cy5 at its N-terminus. Microscopic techniques, in particular cryogenic TEM (cryo-TEM) and cryo-electron tomography technique (cryo-ET), reveal two types of highly ordered three-dimensional assembly structures on the micrometer scale. Unbranched compact layered rods are observed at pH 7.4 and two-dimensional membrane-like assemblies at pH 3.4, both species displaying spectral features of H-aggregates. Molecular dynamics simulations reveal that the coupling of Cy5 moieties promotes the formation of both ultrastructures, whereas the protonation states of acidic and basic amino acid side chains dictates their ultimate three-dimensional organization.

Toward Light-Controlled Supramolecular Peptide Dimerization.

The selective photodeprotection of the NVoc-modified FGG tripeptide yields the transformation of its 1:1 receptor-ligand complex with cucurbit[8]uril into a homoternary FGG2@CB8 assembly. The resulting light-induced dimerization of the model peptide provides a tool for the implementation of stimuli-responsive supramolecular chemistry in biologically relevant contexts.

Approaches to Obtaining Fluorinated α-Amino Acids.

Fluorine does not belong to the pool of chemical elements that nature uses to build organic matter. However, chemists have exploited the unique properties of fluorine and produced countless fluoro-organic compounds without which our everyday lives would be unimaginable. The incorporation of fluorine into amino acids established a completely new class of amino acids and their properties, and those of the biopolymers constructed from them are extremely interesting. Increasing interest in this class of amino acids caused the demand for robust and stereoselective synthetic protocols that enable straightforward access to these building blocks. Herein, we present a comprehensive account of the literature in this field going back to 1995. We place special emphasis on a particular fluorination strategy. The four main sections describe fluorinated versions of alkyl, cyclic, aromatic amino acids, and also nickel-complexes to access them. We progress by one carbon unit increments. Special cases of amino acids for which there is no natural counterpart are described at the end of each section. Synthetic access to each of the amino acids is summarized in form of a table at the end of this article with the aim to make the information easily accessible to the reader.

Risk factors for sick leave in Brazilian oil industry workers.

OBJECTIVE: Studies point to a complex and multifactorial aetiology for sickness absence, but there is still insufficient evidence on how occupational and non-occupational risk factors contribute to this phenomenon. This study aimed to identify occupational and non-occupational factors related to sickness absence in oil industry workers, as well as their population attributable fractions (PAF). METHODS: A retrospective cohort study of 2028 oil industry workers was conducted in the state of Bahia, Brazil, between January 1, 2012 and December 31, 2016. Study data were obtained from workers´ medical register. The dependent variable was the first event of sick leave for five or more days, and the independent variables were gender, age, years of work, type of work activity, level of education, work schedule, awkward working postures, body mass index, smoking, physical activity and alcohol consumption. Cox regression analysis identified risk factors to the outcome and PAF were calculated. RESULTS: Being female (HR 1.79), being older (HR 1.44), non-graduated ones (HR 1.44), being of operational staff (HR 1.18), overweight (HR 1.13), obesity (HR 1.39), smoking (HR 1.61) and former smoking (HR 1.30) were risk factors for sickness absence in this cohort. Of all sickness absence in the study population, 6% could be attributed to overweight, 10% to obesity, and 8% to smoking. CONCLUSION: To prevent sick leave, we recommend programs focused on combating smoking and encouraging healthy habits, aiming at weight control and the prevention of chronic diseases. Population attributable fractions point that a universal approach, aimed at the total population will be more cost-effective than health promotion programmes restricted for high-risk populations.

A new biostimulation approach based on the concept of remaining P for soil bioremediation.

C:N:P ratio is generally adopted to estimate the amount of nitrogen and phosphorus to be added to soils to accelerate biodegradation of organic contaminants. However, differences in P fixation among soils lead to varying amounts of available P when a specific dose of the element is applied to different soils. Thus, the application of fertilizers to achieve a previously established C:P ratio leads to biodegradation rates that can be lower than the theoretical maximum. In this study, we developed an equation to estimate the dose of P required to maximize organic contaminant biodegradation in soils as a function of remaining P (P-rem), using diesel as a model contaminant. The soils were contaminated with diesel and received six doses of P. CO2 emission was used to estimate biodegradation of hydrocarbons. Biodegradation increased with P doses. The P level that provided the highest hydrocarbon biodegradation rate showed linear and negative correlation with P-rem. The result shows that the requirement for P decreases as the P-rem of the soil increases (or the P-fixing capacity decreases). The dose of P recommended to maximize hydrocarbon biodegradation rate in soil can be estimated by the formula P (mg/dm3) = 436.5-5.39 × P-rem (mg/L).

Associations of multisite pain with healthcare utilization, sickness absence and restrictions at work.

PURPOSE: Despite the apparent importance of multisite musculoskeletal pain (MMP) for functioning, there is still a lack of studies that have investigated the influence of MMP on healthcare utilization (HU), sickness absence (SA) and restrictions of work (RW). This study described the HU, SA and RW due to musculoskeletal pain (MP) in different body sites and according to number of pain sites and investigated associations between number of pain sites with these three outcomes in workers from Bahia, Brazil. METHODS: This study was based on two cross-sectional surveys carried out in 2010 and 2012. The response in the pooled data was 97 % (n = 1070, 228 women and 842 men). Interviewer-administered questionnaire was used with questions on HU, SA and RW due to MP. The number of pain sites is the sum score of eight body sites with pain in previous 12 months. Covariates were age, gender, physical and psychosocial work demands, leisure-time physical activities and body mass index. Cox regression models, properly applied to a cross-sectional study, determined the associations between number of pain sites with the three outcomes. RESULTS: Prevalence of MP in the previous 12 months is 81.2 %, and MMP accounted for two-thirds of pain. We found consistently increasing occurrence of HU, SA and RW with increasing number of pain sites. For individuals with pain in four or more body sites, the utilization of health care was 1.7-fold the utilization by workers with single-site pain. Having pain in four or more sites increased the prevalence of SA 3.6-fold and of RW 4.0-fold compared with having single-site pain, after adjustment by covariates. CONCLUSIONS: The functional consequences of pain depend on how much body regions are affected, i.e., the more widespread pain, the higher the likelihood of medical consumption, sickness absence and restricted work. Given the high comorbidity, the number of pain sites, instead of specific body site of pain, seems to be a useful measure to anticipate interventions at workplaces for musculoskeletal disease prevention.

The concurrence of musculoskeletal pain and associated work-related factors: a cross sectional study.

BACKGROUND: Several recent studies have described the presence of musculoskeletal complaints, presenting evidence that multisite musculoskeletal pain (MP) is more often present than single-site musculoskeletal pain. However, less is known about determinants of this multimorbidity, particularly, concerning the role of occupational factors and, mainly, what determines single or multisite pain. This study described the associations between pain in different body sites and investigated related factors to MP in workers from Brazil. METHODS: A total of 1070 workers (228 women and 842 men), from urban cleaning services and from shoe manufacturers, participated in this cross sectional study (response 97 %). Interviewer-administered questionnaire included sociodemographic factors, physical and psychosocial work demands, leisure-time activities and musculoskeletal pain which was presence of pain in previous seven days, considering eight body sites and MP, the sum score of all painful sites, varying 0-8. A factor analysis was performed that captured the nine variables of physical exposure into two latent factors. Associations of pain between different body sites were assessed. Cox regression analyses, presenting the prevalence ratio (PR), showed the related factors to MP. RESULTS: In the previous seven days, 30 % of workers had MP. For all body sites, comorbidity ranged from 72 % to 91 %. Having pain in one body site is associated with pain in other site and the associations between proximal sites were stronger than between more distal sites. High exposure to manual material handling and awkward postures (PR = 1.5, 95 % CI 1.1-2.0), job strain (PR = 1.2, 95 % CI 1.0-1.6), and low social support (PR = 1.3, 95 % CI 1.0-1.7) and being woman (PR = 1.7, 95 % CI 1.3-2.3) were associated with MP. Risk factors for single-site pain and for subsequent musculoskeletal comorbidity were very similar, suggesting an additive effect of risk factors. CONCLUSIONS: Most workers reported MP that was associated with several work-related factors. The findings support the idea that multisite pain is a continuum of single-site pain, maintained by exposure to several risk factors, rather than the result of a specific risk factor that initiates the multisite pain but not single-site pain. Workplace interventions are needed to decrease the number of pain sites, in order to improve the worker's health.

Presenteeism and Low Work Performance Among Fruit Farm Workers in Northeast Brazil.

OBJECTIVES: The "decision" to go to work, or remain absent, when unwell undermines worker energy. I is therefore understandable that low work performance is one of the main consequences of the act of presenteeism. This study evaluated the association between presenteeism, and absolute and relative work performance among fruit farm workers in Northeast Brazil. METHODS: A cross-sectional study investigated 340 paid employees of both sexes, aged 18 years or over, who worked during the 2019 irrigated fruit harvest in the municipality of Petrolina, Northeast Brazil. Absolute and relative work performance were measured using the Health and Work Performance Questionnaire. Workers who answered "one or more days" to the question "How many days have you worked this season when you were injured or unwell?" were considered presenteeists. Poisson regression with robust variance models was used to estimate the prevalence ratios of the main associations, adjusted for potential confounders. RESULTS: Presenteeist workers presented lower absolute and relative work performance than non-presenteeist workers according to the strata for most of the investigated factors. The associations between presenteeism, and absolute (adjusted prevalence ratio = 1.31) and relative work performance (adjusted prevalence ratio = 1.45) remained strong, even after multiple adjustments for other relevant variables. CONCLUSION: Presenteeist fruit culture workers self-assessed their work performance as lower than non-presenteeist ones.

Resistant Escherichia coli isolated from wild mammals from two rescue and rehabilitation centers in Costa Rica: characterization and public health relevance.

This study aimed to characterize the antimicrobial resistance (AMR) and virulence profiles of 67 Escherichia coli isolates obtained from faecal samples of 77 wild mammals from 19 different species, admitted in two rescue and rehabilitation centers in Costa Rica. It was possible to classify 48% (n = 32) of the isolates as multidrug-resistant, and while the highest resistance levels were found towards commonly prescribed antimicrobials, resistance to fluoroquinolones and third generation cephalosporins were also observed. Isolates obtained from samples of rehabilitated animals or animals treated with antibiotics were found to have significantly higher AMR levels, with the former also having a significant association with a multidrug-resistance profile. Additionally, the isolates displayed the capacity to produce α-haemolysins (n = 64, 96%), biofilms (n = 51, 76%) and protease (n = 21, 31%). Our results showed that AMR might be a widespread phenomenon within Costa Rican wildlife and that both free-ranging and rehabilitated wild mammals are potential carriers of bacteria with important resistance and virulence profiles. These results highlight the need to study potential sources of resistance determinants to wildlife, and to determine if wild animals can disseminate resistant bacteria in the environment, potentially posing a significant threat to public health and hindering the implementation of a "One Health" approach.

Precarious work and methodological challenges to study hard-to-reach populations.

OBJECTIVE: To describe the methodological challenges and strategies of a web survey on the working conditions and health among delivery workers. METHODS: The study population consisted of Brazilian delivery workers operating in the national territory. Procedures include building solid and ongoing collaboration with worker representatives and conducting a four-month data collection from February to May 2022, sharing the link to the online questionnaire on social media such as social networks (Facebook, Instagram) and messaging apps (WhatsApp, Telegram). RESULTS: The recruitment of 41 leaders or influencers of delivery workers increased the dissemination of the study, some of whom participated in the consensual validation of the questionnaire; the production of content for social media for the dissemination of the questionnaire link on social networks and applications, and the in-person dissemination of the study at the delivery workers' meeting points during the workday played a fundamental role, totaling around 132 hours in 45 shifts. The strategies adopted for data collection with a hybrid approach to dissemination made it possible to carry out the web survey. After four months of the web survey, 564 delivery workers, 543 men and 18 women, responded to the online questionnaire. CONCLUSION: The web survey presented methodological strategies to overcome the challenge of reaching workers, including hybrid work, to increase the participation of workers, on whom epidemiological research is still scarce.