Mobile mapping technologies, based on techniques such as simultaneous localization and mapping (SLAM) and surface-from-motion (SfM), are being vigorously developed both in the scientific community and in industry. They are crucial concepts for automated 3D surveying and autonomous vehicles. For various applications, rotating multiline scanners, manufactured, for example, by Velodyne and Ouster, are utilized as the main sensor of the mapping hardware system. However, their principle of operation has a substantial drawback, as their scanning pattern creates natural gaps between the scanning lines. In some models, the vertical lidar field of view can also be severely limited. To overcome these issues, more sensors could be employed, which would significantly increase the cost of the mapping system. Instead, some investigators have added a tilting or rotating motor to the lidar. Although the effectiveness of such a solution is usually clearly visible, its impact on the quality of the acquired 3D data has not yet been investigated. This paper presents an adjustable mapping system, which allows for switching between a stable, tilting or fully rotating lidar position. A simple experiment in a building corridor was performed, simulating the conditions of a mobile robot passing through a narrow tunnel: a common setting for applications, such as mining surveying or industrial facility inspection. A SLAM algorithm is utilized to create a coherent 3D point cloud of the mapped corridor for three settings of the sensor movement. The extent of improvement in the 3D data quality when using the tilting and rotating lidar, compared to keeping a stable position, is quantified. Different metrics are proposed to account for different aspects of the 3D data quality, such as completeness, density and geometry coherence. The ability of SLAM algorithms to faithfully represent selected objects appearing in the mapped scene is also examined. The results show that the fully rotating solution is optimal in terms of most of the metrics analyzed. However, the improvement observed from a horizontally mounted sensor to a tilting sensor was the most significant. Simultaneous localization and mapping (SLAM) represents one of the most significant computational problems for any 3D reconstruction. It can be applied to autonomous cars and robotics inspection missions to solve specific tasks, in particular map generation and obstacle detection. The most promising data source for performing these tasks is application of the lidar distance measurement method. Although many 3D-mapping lidar systems have been developed and are used in various applications, the quality and accuracy of the generated 3D point clouds and 3D geometries often remain insufficient, particularly in complex or unstable environments. However, one of the most rapidly growing fields of application for mobile mapping systems is underground mining, as the speed and possible automation of performing such measurements enables the rapid acquisition of dense 3D data for vast underground structures. For these reasons, constant improvement is needed in measurement system construction and point cloud data processing. New approaches to the analysis and methodological assessment of the 3D data quality are also required. One promising solution, improving the hardware side of the mapping system, is the design of a spinning lidar sensor with an additional module that rotates the sensor around another axis. This actuated lidar design, in which a scanner is combined with an actuation mechanism to scan a 3D volume rather than a single line, has already been used in numerous mobile robot SLAM applications .Therefore, the main objective of our study was to quantitatively and qualitatively assess the influence of a well-known approach using lidar rotation on the accuracy and value of a 3D point cloud using an inspection robot in a test environment mimicking an underground mine tunnel. The paper is structured as follows: first, in The data acquisition strategy for autonomous vehicles and inspection robots using the lidar system needs to be based on an understanding of specific environmental parameters, such as the presence of a variety of dynamic or static obstacles, ensuring the highest possible level of accuracy in the scanned data. Measurement systems with rotating lidar sensors enable the capture of dense and close-to-spherical data about the surrounding environment . Thus, nThe design of a dual 2D/3D lidar mapping system and a six-degrees-of-freedom (DOF) interpolated odometry, where 2D lidar is used to enhance 3D solid-state lidar performance used on a ground vehicle platform, can be found in . The desConstruction of the scanning actuation mechanism can have an impact on different positions of the rotation center of the lidar mirror and construction itself; thus, sufficient calibration methods are needed. The description of an automatic calibration method for the actuated lidar can be found in . An extePoint cloud accuracy and data-quality assessment are amongst the most important factors for the creation of reliable, error-free methods used in laser scanning. To tackle the recurrent problems of misregistration, outlier detection and over-completeness, a comparison of several methods is needed. In , the useWith regard to the underground mining environment being investigated in the present study, SLAM solutions and applications in real or artificial spaces need to be considered. Underground workings are characterized by significant irregularities in the surrounding geometries and by dustiness that can affect the performance of the mobile system and the quality of measurements obtained. The results presented in the studies referred to below confirm that SLAM-enabled laser scanning represents a promising method for underground mining tunnel mapping and examination. One of the first studies that indicated the potential for high-resolution 3D-mapping of an underground mine involved a cart-mounted 3D-laser-scanner setup and an automatic 3D-modeling method . SimilarThe authors in  proposedThe literature review highlighted that there are still gaps to be filled with respect to evaluation methods for comparing the results  produced with different SLAM systems, with respect to both the hardware and the software used. Developing such methods would assist in choosing suitable solutions for selected use-case environments, optimizing the cost of scanning, system complexity and data size and quality. This study aims to tackle this challenge.The purpose of developing an adjustable mapping system for this study was to enable additional, controlled rotation of the lidar device around its longitudinal axis. In consequence, the effective field of view (FoV) of the sensor can be gradually increased, almost up to the point of a full-spherical FoV (excluding occlusions caused by the sensor mounting and the robot). The adjustability of the proposed system is controlled by the operator, who can easily change the range of lidar rotation using control software options on the remotely connected tablet. In this research, we utilized these options to emulate three different strategies for lidar head mounting, which have been used in various commercial solutions and research prototypes. The details of the system configuration are presented in the following paragraphs.In this study, a 16-line Velodyne VLP-16 lidar is used. The sensor is mounted on a rotating module, mounted above the set of sensors designed for inspection purposes A block diagram of the data-acquisition system with the actuator module is shown in In this study, several steps of point cloud acquisition are taken to obtain coherent and noise-free global point clouds. The aim is to ensure the highest quality of results from the different methods tested so that the comparison outcomes would not be affected by external factors or imperfect execution and repetition of the experiment. The parameter values in each step are universally chosen and kept consistent for each case. Although their method-specific tuning could potentially improve the accuracy of results, it might significantly influence the data density and, thus, introduce bias into one of the most important aspects of the analysis.tf broadcaster [First, the robot trajectory is estimated in real time with the SLAM system, consisting of A-LOAM lidar odometry , scan coadcaster , which radcaster ,29, withAn important part of the applied SLAM algorithm is correction for motion distortion of point clouds. The lidar used in the study acquires 360° 3D point clouds in constant motion, revolving internally around its z-axis. One revolution (a sweep) takes 0.1 s and an aggregated set of point coordinates is sent by the sensor to the PC. However, when the scanner is in motion, the points acquired between the start and the end of the sweep have a slightly different frame of reference. This results in aggregating in each point cloud points acquired from slightly different positions, introducing a systematic error into the measurements . Since tLidar odometry provides a quick and constant, but preliminary, sensor pose estimate. It is susceptible to long-term drift, especially with respect to orientation in the 3D space. With a rotating sensor, such as that used in the study, this could cause unacceptable mapping results, with common errors occurring, such as double walls or rotated corridors. To prevent this, loop closure detection and pose-graph optimization are included in the software system. In our test measurements, we did not have explicit loop closures in terms of returning to the same place the robot visited earlier. Therefore, Scan Context++ was used to additionally bind the trajectory after each full resolution of the actuator. The point clouds acquired at the same actuator inclination angle are/were similar due to the low speed of the robot. The resulting matches between scans are added as a constraint to the pose graph and optimized with GTSAM, reducing the drift of the odometry algorithm.Then, clusters of points representing dynamic objects are removed in post-processing using removert . This stEvaluating the quality of 3D point cloud data acquired with a mobile mapping system is not a straightforward task. Many metrics have been proposed and used to assess the accuracy of SLAM measurements, such as the absolute trajectory error (ATE), the relative trajectory error (RTE) and the relative position error (RPE) ,36. HoweN—number of neighboring points in the radius R around the analyzed point.One of the metrics that can be used to estimate the local quality of a point cloud is its density. This simply describes the number of measured points per chosen unit of reference: a volume or a surface. For this study, since the mapped area primarily contains approximately planar surfaces, surface density was calculated with respect to Equation  and analData density provides information about the spatial distribution of points and their number, characterizing the redundancy of the measured geometry. However, this metric does not convey information on the noise level of the analyzed point cloud. To address this, another component was introduced to the analysis, namely, object reconstruction quality, that is, how accurate the measured geometry is of a single object in the scanned scene.For evaluating the object reconstruction quality, i.e., the local consistency of a point cloud, several objects were chosen as samples. They were manually identified in each point cloud and compared between different tested lidar SLAM configurations. For planar objects, such as walls, doors or floor, surface variation ,39 also. This meIn the cases of two distinctive objects in the study area, namely a ceiling lamp and a trashcan, such analysis would not be meaningful since their geometry is more complex than a plane. To account for this, for all objects, point cloud resolution was additionally compared by listing the number of points per object.The last analyzed aspect of the point clouds acquired with SLAM is their completeness. Since the raw number of points is heavily influenced by noise and redundant measurements, other metrics need to be utilized for this purpose. Such a metric should describe a unique volume of space, containing any measured points. Two data structures commonly used in 3D-data processing have this property: a voxelgrid and an octree. Both divide a 3D space into a regular volumetric grid of boxes. For the voxelgrid, the size of a single voxel is fixed, and an octree contains a multiresolutional representation of the scene, sequentially dividing cells at each level into octants ,43. To aSensor in fixed horizontal position, i.e., horizontal lidar;Sensor rotating in the full range from −90° to 90°, i.e., rotating lidar;Sensor rotating in the limited range from −45° to 45°, i.e., tilting lidar.Several experimental data acquisitions were performed with a wheeled mobile robot  to (f), in To begin with, the reconstruction quality of objects (a), (c), (e), and (f) was investigated, since they consist mostly of planar objects. Consecutively, they are: a vertical surface located high, a vertical surface at the same level as the measurement system, an area of the floor at the mid-section of the measurement area and a door at the side wall in the corridor. Their point clouds, colored by the calculated surface variation per point, are presented in Local accuracy of the final point clouds was assessed using a least-squares plane fit for objects (a), (c) and (e). The results are listed in Objects (b) and (d), which had more complex geometries, were analyzed only in a simplified context. Their point clouds are shown in The completeness of 3D data in the context of object reconstruction, as well as the point cloud density, can be additionally summed up with a simple metric—the point count per object. Such an overview is presented in Different aspects of the 3D-data quality of three common hardware solutions utilizing a 3D lidar scanner for the SLAM problem were investigated. Multi-metric comparison was conducted to analyze factors such as local surface density and variation, plane reconstruction accuracy and numbers of octree cells, voxels and points per mapped object. This analysis enabled us to obtain insights into the behavior of SLAM in tunnel-like conditions, especially with respect to key aspects of inspection and mapping robotic missions in constrained, underground environments. Similarly to the method described in , in the Increasing the complexity of the system through introduction of an actuator to rotate the spinning lidar around another axis greatly increased the data density and completeness, and did not negatively impact the point cloud local coherence. Although the sensor rotating in its full range generally obtained the best results, a tilting sensor achieved results that were not much worse and provided significant improvement over the static, horizontal placement of the lidar. Depending on the metrics analyzed, generally, the performance of the rotating lidar was from 35% to almost 50% better than that of the tilting lidar. The tilting lidar obtained a smoother data density distribution and almost 200% better completeness (based on voxelgrid and octree cell counts) than the horizontal lidar, while still maintaining comparable plane fit accuracy and mean data density. Choosing the right tool for the selected measurement site will depend on the dimensions of the structure, notably its height and width. The presence of objects causing numerous occlusions, common for underground mining environments, would also favor the selection of one of the actuated lidar mounts.During inspection missions in underground mines, a massive amount of 3D data is collected with SLAM to be used for both navigation and 3D analysis. In the case of the latter, completeness of different object representations in the point cloud is crucial to enable machine learning algorithms to perform classification successfully and to correctly distinguish different objects of interest, which can then be processed with specialized, use-case-targeted algorithms. Therefore, given the results presented in


As the number of children and young people (CYP) presenting to Emergency Departments (ED) with acute mental health (MH) presentations has been steadily increasing over the years and further accelerated by the pandemic, there is an urgent need to develop and evaluate innovative solutions to respond to this growing challenge.The evaluation of the Safeguards Teams Program (STP) aims to ascertain the impact, implementation and economic analysis of this acute rapid response recovery-focused, trauma-informed assessment and brief intervention for CYP (aged 0–17 years) presenting in acute MH crisis and their families/caregivers.The STP will support consumers (patients) and their families/caregivers to navigate the complex and often fragmented child and adolescent MH services (CAMHS) landscape, thereby avoiding unnecessary ED presentations or hospitalisations, and facilitating comprehensive assessment and appropriate care pathways for those who present in crisis.The STP is expected to provide CYP in MH crisis and their support networks with early access to evidence-based specialist care at the right place and time.Implementation of the STP will assist with identifying and addressing gaps in acute care for CYP and provide the necessary evidence for service redesign in collaboration with consumers, service providers and other stakeholders. Around one in seven 15%) children and adolescents globally are diagnosed with mental health (MH) conditions 5% childr3. MH conAs the number of children and young people (CYP) presenting to ED with acute MH presentations increases, there has been an attempt to find alternative solutions to manage crises within the community setting for vulnerable children and young people (CYP). The literature reports varied approaches to specific interventions, such as Multisystemic Therapy for behaviour disorders (4), Assertive Community Treatment for high-risk and difficult to engage adolescents who have a severe mental illness (5), and ‘one-stop shops’ of Integrated Community-Based Youth Service (C-BYS) Hubs that aim to cater for psychosocial, educational, physical and MH needs in an early intervention model (6). However, these models require specific funding, strong collaboration, and integration of services.The approach to acute crisis presentation requires accessibility and availability outside of usual business hours, outreach into the community, and individually tailored care that is specific for CYP and their families while also ensuring that appropriate referrals and linkages with other relevant services are in place to support CYP consumers following a crisis intervention. Without appropriate post-intervention support, CYP with MH conditions are highly likely to have poor health and social outcomes and high ongoing service user needs. Failing to support CYP during the crisis and adequately supporting them thereafter is expected to have a significant adverse impact on their long-term trajectory that may include poor adult MH, low school engagement and performance, unemployment, high dependence on welfare, unstable housing conditions, involvement with the child protection system, criminal activity, drug and alcohol dependency, and premature death .A recent systematic review found evidence for the effectiveness of crisis interventions for CYP who are in acute MH distress via reducing ED presentations and hospitalisations and reducing health system costs, but the review also highlighted the scarcity of studies in this area and called for further studies to bridge this knowledge gap . Here weThe Safeguards Teams Program aims to improve access to timely, evidence-based, recovery-focused, and trauma-informed assessment and brief intervention for children and adolescents presenting in acute MH crisis and their families/caregivers.The STP will reduce the rates of CYP re-presenting to the ED with acute MH concerns including the use of emergency services and/or necessitating re-hospitalisations and result in better engagement with services (primary outcome), and leading to better mental health outcomes, thereby minimising school (better school attendance) and psychosocial (better engagement and participation) disruption (secondary outcomes).An additional hypothesis is that the implementation of the STP program will be feasible, acceptable, and accessible and will result in effective referrals and linkages to relevant services post-intervention with the uptake of these recommendations and service satisfaction for consumers (CYP and their families), service providers  and other stakeholders . An economic evaluation will also be undertaken to ascertain the value for money by comparing the incremental cost and benefit based on child and family outcomes and service experiences. Return on investment, associated with wide-scale implementation, will be modelled based on the trial outcomes.The study will utilise a pre- (Time 1/baseline) and post-intervention (Time 2) design to compare outcomes of individuals who receive STP at entry into the program and then on exit from the STP followed by data collection at 3 months post-intervention (Time 3) for re-presentation to ED or in-patient wards including the use of emergency services  and engagement with referred services and uptake of recommendations. In addition to questionnaire/survey data collected at the three-time points, EMR data will be collated for those who participated in the STP program and from another CAMHS service site where CYP presenting in acute crisis received care as usual. Further qualitative study using focus groups and in-depth interviews, and ethnographic approaches, such as observations and reflections will be completed with consumers (CYP and their parents/caregivers), service providers and relevant stakeholders. An implementation evaluation of the overall impact, implementation process and economic analysis will also be carried out.The sample for the STP will include children and adolescents 0–17 years who are experiencing an acute MH crisis, who are not currently undertaking MH treatment and would benefit from a rapid response MH service. Services will be provided for both CYP and their parents/caregivers.Children and adolescents aged 0–17 years presenting in an acute mental health crisis , for an urgent or immediate response)  index).Mental health outcomes will be ascertained using pre- (at admission) and post- (at discharge) questionnaire surveys completed by clinicians, CYP and their parents/caregivers . QualitaA semi-structured questionnaire has been developed to gather not only baseline demographic data but also information regarding how CYP functions  relative to mental health service access; attendance, participation and performance at school; participation in sports or other physical or social activities; and the current relationship between the CYP and parent(s), other family members , friends, and social support network connections.Data will be collected at each time point.NSW Mental Health Carer Experience Survey provides a better understanding of the experiences, needs and outcomes of carers. The measure consists of 37-items.Children’s Global Assessment Scale (CGAS) scale is a clinician-rated scale used to rate the functioning of children and young people aged 6–17 years old related to their psychological and social functioning. It is a brief- 10-item observer-rated scale.Clinical Global Impression – Severity scale (CGI-S) and Improvement scale (CGI-I) comprise a brief- 3-item observer-rated scale that requires the clinician to assess the overall severity of the individual’s illness and overall improvement in the individual’s illness relative to a baseline state.Health of the Nation Outcome Scales for Children and Adolescents (HoNOSCA) is a clinician-rated measure. The first section consists of questions related to the different types of problems that a child or adolescent might be experiencing; the second part of the survey relates to the parent/caregiver’s knowledge of the nature of their child’s difficulties and information about MH services available to support the child and the family.Kessler Psychological Distress Scale (K10) explores questions about depressive symptoms that CYP may have experienced. The survey is a 10-item measure and explores distress based on questions about anxiety and depressive symptoms that a person with a MH disorder may experience within a 4-week period.Pediatric Symptom Checklist—Youth Report (Y-PSC) is a 17-item measurement psychosocial screen designed to facilitate the recognition of cognitive, emotional, and behavioural problems completed by youth aged 12–17 years and up. Similarly, the parent-completed Pediatric Symptom Checklist is also a 17-item measurement tool and provides a measure of their children’s psychosocial functioning.Pictorial Pediatric Symptom Checklist (PPSC) is a pictorial psychosocial screen designed to facilitate the recognition of cognitive, emotional, learning, and behavioural problems completed by children and youth aged 6–11 years and up. It is a brief 35-item measurement tool.Your Experience of Service (YES) questionnaire explores the experiences of consumers about both their current and previous use of MH services.A mix of qualitative, quantitative, and ethnographic approaches will be used to examine the effectiveness of the implementation of the STP with a focus on key implementation outcome metrics including acceptability, appropriateness, fidelity, feasibility, coverage, cost, and sustainability as detailed in The STP processes of how care pathways, practices and agreements have been established and developed from ED, mental health services and community-managed organisations for children and adolescents with acute mental health crises will also be evaluated using a mixed-methods approach. This includes exploring how clinical practice changes have occurred within ED, mental health, and community-based organisations to prevent or decrease avoidable hospital ED presentations and hospital admissions . The data collection source will include interviews and focus groups with mental health clinicians, primary care clinicians, hospital clinicians, government agency representatives, community-managed organisations/stakeholders, and ethnographic observations in STP multidisciplinary meetings and stakeholders/steering committee meetings.Quantitative analysis: The effectiveness of the Safeguards Teams program will be evaluated by examining the difference in outcomes on entry before participants enrol into the program, and on exit, following a Safeguards encounter, using appropriate statistical methods for different outcomes. Descriptive analyses will explore changes following Safeguards Team’s service delivery. We will use paired samples t-test to compare pre- versus post-intervention outcomes as per measures outlined earlier. Additionally, percent change at entry and exit from the STP from baseline for efficacy variables, will be analysed using analysis of covariance (ANCOVA) with respective baseline value as covariate. Least-square means (LSM) and 95% confidence intervals will be evaluated from the ANCOVA. The data of percent changes will be assumed as normally distributed.Qualitative analysis: In-depth semi-structured interviews of consumers and service providers (peer-workers and clinicians) will be conducted using the Consolidated Framework for Implementation Research (CFIR) [h (CFIR)  frameworh (CFIR)  for datah (CFIR) .Economic Evaluation: The economic evaluation will aim to understand the cost-effectiveness of the Safeguards Program to inform future decision-making. Trial-based economic analyses will be conducted to ascertain value for money by comparing (i) the incremental cost associated with the STP, including implementation costs, and (ii) incremental benefit assessed according to patient-reported outcome and experience measures and clinical measures. Return-on-investment, associated with wide-scale implementation, will be modelled based on the trial outcomes. Dynamic Simulation Models (DSMs) will be used to identify optimal combinations of services and referral trajectories that will provide policymakers and clinicians with practice-specific information to inform clinical decisions and forecast the potential impacts of services on outcomes. The evaluation will consider the economic return on investment as well as social return on investment.The study has been approved by the University of New South Wales Human Research Ethics Committee, reference HC220192, and SWSLHD Human Research Ethics and Governance, reference 2022/ETH00808.In most cases, little is known about the relative effectiveness and efficiency of different CAMHS delivery models . One reaIt is expected that the STP will provide CYP with MH conditions and their support networks with early access to comprehensive assessment and evidence-based acute response and referral and linkages to appropriate care delivered by specialist clinicians and support services. Outcomes from the STP may also provide clinicians involved in CAMHS with: (1) a more robust approach to service redesign specifically tailored for the provision of MH care and services to CYP consumers and their families experiencing acute MH problems; (2) data to support an economic evaluation to determine the cost-effectiveness of the Safeguards program compared with treatment as usual to inform future decision-making; (3) a mixed methods approach to explore any improvements regarding engagement, access, and uptake/integration of care in mental MH with the inclusion of the Safeguards Teams within the CAMHS as well as clinical practice changes within ED, MH services and community-based settings to prevent or decrease avoidable hospital ED presentations and hospital admissions .Moreover, the Safeguards Program will draw on building connections with local stakeholders and improving service delivery across CYP. As a result of embedding the new Safeguards Teams into the existing CAMHS structures, it will also ensure that access to MH care is based on level of acuity, risk, complexity, and distress, and is not limited by diagnosis or co-morbidities.Outcomes associated with the impact, implementation, and economic evaluation of the STP are expected to assist with identifying gaps in acute care for children and adolescents in the community and provide an alternative model of care for CYP presenting in MH crisis.


Introduction: Ablation therapy is a commonly used tool in the management of hepatocellular carcinoma (HCC). After ablation, dying cancer cells release a variety of substances that trigger subsequent immune responses. Immunogenic cell death (ICD) has been a trending topic in recent years and has been discussed many times along with oncologic chemotherapy. However, the subject of ablative therapy and ICDs has been little discussed. The purpose of this study was to investigate whether ablation treatment induces ICD in HCC cells and whether different types of ICDs arise because of different ablation temperatures.Methods: Four different HCC cell lines  were cultured and treated under different temperatures . Cell Counting Kit-8 assay was performed to analyze the viability of different cell lines. Apoptosis was detected by flow cytometry assay, and a few ICD-related cytokines  were detected by immunofluorescence or enzyme-linked immunosorbent assay.Results: The apoptosis rate of all kinds of cells increased significantly in −80°C group (p < 0.01) and 60°C group (p < 0.01). The expression levels of ICD-related cytokines were mostly significantly different between the different groups. For calreticulin, Hepa1-6 cells and SMMC7221 cells showed significantly higher protein expression levels in 60°C group (p < 0.01) and significantly lower protein expression levels −80°C group (p < 0.01). The ATP, high mobility group box 1 and CXCL10 expression levels were significantly higher in 60°C, −80°C and −40°C group of all four cell lines (p < 0.01).Conclusion: Different ablative treatments could induce different types of ICDs in HCC cells, providing a promising track for the development of individualized cancer therapies. There ahibition  or promohibition  of a disTo unravel how the immune system deciphers the complex signals sent by dying cancer cells, considerable efforts have been devoted. Notably, a precise combination of damage-associated molecular patterns (DAMPs) presented in a spatiotemporally orchestrated sequence by dying cancer cells can bolster the immune reaction that drives specific T-cell immunity, thereby implementing durable tumor regression and facilitating immunologic memory .To distinguish the different forms of dying tumor cells, a novel concept of immunogenic cell death (ICD) was introduced. Through concerted expression and release of a broad array of DAMPs, this specific form of apoptosis can lead to a pro-inflammatory immune process in an immunocompetent host . Relatedin vitro cultured HCC cells, we aimed to answer these two questions: (1) whether heat ablation or cryoablation, sparks ICDs in HCC cells, (2) whether the different temperature sparks different ICDs. We cultured two murine HCC cell lines (H22 and Hepa-16), and two human HCC cell lines (HepG2 and SMMC7221). Then we subjected them to acute cryoablation or heat ablation at different temperatures , with 37°C serving as a control group. Cell apoptosis was measured by flow cytometry, and the expression levels of various ICD-related cytokines were detected by immunofluorescence or enzyme-linked immunosorbent assay. Our data suggested that in different HCC cell lines, different treatment induced different expression levels of ICD-related cytokines.In recent years, a series of articles have explored whether the commonly used oncologic chemotherapeutic agents in clinical practice carry the ability to trigger ICDs , while t2 atmosphere.The human HCC cell lines , and the mouse HCC cell lines (H22 and Hepa1-6) were kindly provided by Stem Cell Bank, Chinese Academy of Sciences. The cells were grown and maintained in RPMI-1640 medium  containing 10% fetal bovine serum  and antibiotics , and the culture was incubated at 37°C in a 5% COFor each cell line, cells were divided into five groups for different thermal-treatment, −80°C, −40°C, 0°C, 37°C, 60°C. For cryoablation group, cells were placed in a refrigerator at −80°C, −40°C, or 0°C for 30 min. For heat ablation group and control group, cells were heated in a 60°C or 37°C water bath at for 30 min.5/mL per well into 96-well culture plates. After 24 h of incubation, the cells were subjected to the above-mentioned cryoablation or heat ablation. Then, the plates were rinsed, added 10 μL of enhanced CCK-8 reagent  per well, and then incubated for 4 h. Lastly, the 96-well plates were scanned at a wavelength of 490 nm using a microplate reader, and cell survival rate was calculated.Viability of different HCC lines following freezing or heating was analyzed by CCK-8 assay according to the manufacturer’s protocols. In brief, HCC cells were cultured at a concentration of 5 × 105 resuspended cells of each cell line were centrifugated and gently resuspended in 195 μL Annexin V-FITC binding buffer. Next, cells were stained with 5 μL Annexin V-FITC and 10 μL propidium iodide. After 20 min of incubation at 4°C in darkness, the percentage of apoptosis rate was determined using flow cytometry .Flow cytometry was performed using a FITC-Annexin V and PI Apoptosis kit  according to the instruction. In brief, cells were collected after the ablation treatment and resuspended with cold PBS buffer and counted. 1 × 10To evaluate whether cryoablation or heat ablation could induce the predicted emission of specific cytokines, the levels of CXCL10, HMGB-1, and ATP were quantified with commercial ELISA kits  in accordance with the manufacturer’s instructions. For each cytokine, required number of microplate strips were taken out from the matching ELISA kit, and the rest were sealed and stored at 4°C. The set standard wells were spiked with different concentrations of standards, 50 μL each. In the set sample wells, 10 μL of the sample was added firstly, and then 40 μL of sample dilution. No reagents were added to the set blank wells. Next, 100 μL of horseradish peroxidase (HRP)-labeled detection antibody was added to each well, except for the blank wells. The top of the wells was covered firmly with a seal until the end of a 60-min incubation. Then the wells were washed for 5 times and treated with 50 μL of substrate. And after a 30-min incubation in the dark for color development, each well was applied with 50 μL stop solution. The optical density was read at 450 nm using a Multiskan™ GO detector system.The expression of CRT was detected by immunofluorescence assay. Different HCC cells were grown in 24-well plates and underwent cryoablation or heat ablation. After 3 washings with PBS and a 10-min fixation with 4% paraformaldehyde, each well was permeabilized by 200 μL of 0.2% Triton X-100 for 10 min. Next, 5% BSA was added to each well for 1 h to block non-specific staining. Then the primary antibody (Sigma-Aldrich) at 1:400 dilution was applied to the wells, which were stored at 4°C overnight. Followed by 4 washings, the wells were applied with Alexa Fluor 488-labeled goat anti-rabbit IgG (H + L) antibody  at 1:400 dilution, incubated at room temperature for 1 h and washed 4 times. Finally, 150 μL of DAPI was added to each well. After a 4-min incubation at room temperature, the plates were washed with PBS for the last 4 times. The slides were examined under a fluorescence microscope, and the fluorescence intensity of each sample was recorded.p-value <0.05 was considered a statistically significant difference.Dates are presented as the mean ± standard deviation (SD). The statistical analyses were performed using SPSS 25.0 software  and figures were made by GraphPad Prism version 9 . Comparison between different groups was determined by one-way ANOVA with Turkey’s multiple comparison test. in vitro cultured HCC cells to ablation at different temperature is concluded in p < 0.01) and 60°C groups (p < 0.01), while no difference was observed as between −80°C and 60°C groups. The viability of human HepG2 HCC cells decreased after treatment, but the viability of SMMC7721 cells decreased obviously only in −80°C group (p < 0.01), not in −40°C group. And the cell viability (SMMC7721 and HepG2) in −80°C group was much lower than 60°C group (p < 0.01).Cell viabilities of different HCC cell lines were measured by CCK-8 method. The response of p < 0.01) and 60°C group (p < 0.01). And for SMMC7721 and HepG2 cells, the apoptosis rate in −80°C group was much higher than 60°C group (p < 0.01).Furthermore, an apoptosis assay was performed by co-staining with annexin V and PI. Flow cytometry was utilized to measure the apoptosis rate of H22, Hepa1-6, SMMC7721, and HepG2 cells. As shown in p > 0.05). Hepa1-6 cells and SMMC7221 cells showed significantly higher protein expression levels in 60°C group (p < 0.01) and significantly lower protein expression levels in −80°C group (p < 0.01). HepG2 cells showed no significant changes in protein expression levels under 60°C conditions, but significantly lower protein expression levels in −80°C group (p < 0.01).The expression levels of CRT were detected by cellular immunofluorescence assay. The results are shown in p < 0.01). The increase in ATP concentration in Hepa1-6 cells was greater in −80°C group than in 60°C group (p < 0.01). The other three kinds of cells showed higher levels of ATP concentration in 60°C group than in −80°C group (p < 0.01).The ATP and HMGB1 expression levels were detected by commercial ELISA kits. As shown in p < 0.01), and the concentration was a little higher in 60°C group than in −80°C group.As shown in p < 0.01) and thermal treatment (p < 0.01). For H22 and Hepa1-6 cells, the concentrations of CXCL10 were comparable. For SMMC7221 and HepG2 cells, the concentration of CXCL10 was a little higher in 60°C group than in −80°C group.in situ, so as to boost the immunogenicity of the tumor by exposing a broad array of neoantigens. In addition, numerous intracellular components that act as co-stimulatory molecules for immune activation are released post-ablation. Increasingly, efforts are being made to integrate the immune system with thermal ablation , 50°C is essentially high enough to cause coagulative necrosis in no time. Cryoablation arrests viability of local cells by removing heat. It has been suggested by a number of studies that temperatures below −20°C can effectively destroy cancer cells. Researchers generally set the lethal temperature at −40°C, although cryoablation can reduce the interstitial temperatures to as low as −160°C. The extent of cellular damage caused by heat-based ablative therapies depends on three factors: the amount of energy applied, the rate of energy delivery, and the target tissue’s intrinsic thermal sensitivity . Our stuWithin the endoplasmic reticulum, CRT, a soluble protein, carries out several important tasks, which include regulating calcium homeostasis, and assisting in protein folding. When present on the surface of the cell membrane, CRT primarily involves in antigen presentation. CRTs translocated to the membrane of dying cells and distributed in clusters interact with CD91 receptors in phagocytes, which provoke phagocytosis of dying cells  and the in vitro evidence for that HMGB1 promotes HCC cell proliferation, migration, and invasion  and receptor for advanced glycation endproducts (RAGE) and promotes the release of more inflammatory cytokines through the activation of NF-κB pathway . As confinvasion . In thisin vitro, however, normalizing the level of extracellular ATP to the percentage of dead cells helps to distinguish ICD from other types of cell death.It is well known that ATP is the most important energy supplier for all intracellular life activities. Normally, in addition to being present at high levels within the cell, ATP is present at a low level extracellularly. However, under certain stimuli, ATP would be released in excess extracellularly and trigger a series of responses, such as recruitment and transformation of myeloid cells, by binding to and activating purinergic receptor P2Y2 (P2RY2) on the myeloid cell surface . To put Chemokines are secreted proteins structurally similar to cytokines and play an important role in regulating the migration of immune cells and the proliferation of tumor cells, including CXCL10, which possess a strong pro-inflammation capacity. Two recent reports have shed light on the connection between CXCL10 and immunotherapy. One of them demonstrated that CXCL10 can be produced not only by immune cells but also by melanoma cells after the application of immunotherapy, which is a prognostic benefit for the tumor . The othin situ that could boost anti-tumor immunity. Solely inducing tumor cell ICDs to release specific antigens is not successful in triggering an effective immune response most of the time. Under immunosuppressive conditions, additional APCs need to be recruited and activated to counteract the relatively insufficient release of DAMPs (in vivo should accelerate the development of personalized anticancer therapies and identification the optimal combination of treatments for the clinical management of cancer.Given the current clinical treatments, the high risk of relapse and acute or chronic liver function impairment after HCC treatment can hardly be addressed. The development of a low-risk and long-term effective treatment for HCC that can fight against the intrinsic immune privilege of liver remains a great challenge, the fulfillment of which depends on further exploration of the immunological features in the tumor microenvironment. In this direction, one of our ultimate goals is to implement an effective vaccine of DAMPs . As thisin vitro level, and no in vivo experiments have been performed yet. This is mainly owing to the fact that we could not yet heat or cool the target tissue to a specific temperature in vivo. It requires further technological development to break through these limitations.Limitations of this study should be mentioned. Firstly, we used water baths and refrigerator storage to simulate heating and cryo-ablation at different temperatures, which did not fully comply with the specifications of clinical ablation operations. Ideally, we should use a needle antenna to deliver energy, and precisely control the temperature. Secondly, our experiments were limited to the


The importance of animal welfare is being recognized worldwide. Recently, the increasing demand for enhanced laboratory animal welfare has led to clinically featured transformations of animal research institutes. This study aims to describe the process and findings of veterinary medical check-ups and its influence on laboratory dogs and pigs welfare. Regular medical checkups were conducted by the attending veterinarian twice a year to ensure the health and welfare of dogs and pigs in our animal research institute. Based on the findings from the medical checkup, we assessed the current health of dogs and pigs,providing reasonable treatments to prevent the risk of complications.Blood tests and physical examinations revealed clinically relevant findings. Some of these findings were due to insufficient postoperative care after invasive surgical experiments and the remaining were predictable side effects after surgical experiments. However, one finding involved severe gum bleeding due to retained deciduous teeth. This animal was euthanized because it was judged to reach the humane endpoint. Majority of the dogs and pigs at our animal research institute were considered to be healthy, based on the comprehensive results of the medical checkups.Regular medical checkups by the attending veterinarian established enhanced animal welfare, ensuring the accuracy and reproducibility of animal studies. This pioneering veterinary animal care program can serve as a potential advanced guideline for animal research institutes to improve dogs and pigs welfare.The online version contains supplementary material available at 10.1186/s42826-023-00170-7. With progress in the healthcare industry and advanced technologies for the development of new drugs or medical devices, the number of animal research institutes have also increased worldwide. Animal research aims to provide scientific evidence prior to early feasible testing in humans and human clinical trials to assure the safety and efficacy of novel medical device technologies and therapies for humans and animals . A well-Laboratory animals, once considered only a biological reagent, are now being raised in animal research institutes, owing to the emerging global importance of ethical considerations , 4.The welfare of laboratory animals in South Korea is regulated by the ANIMAL PROTECTION ACT with the principles of the 3Rs . The law emphasizes the importance of animal health by stating that “Animal testing shall be conducted in consideration of the enhancement of welfare of humankind and the dignity of lives of animals”. In addition, all animal research institutes should configure an Institutional Animal Care and Use Committee (IACUC) to ensure lawful compliance with the 3Rs .Since physiological indicators of laboratory animals vary depending on their environmental and health conditions, psychological stress or physical distress is considered as a potential trigger to substantial scientific and humane implications for the use of laboratory animals . The staAttending veterinarians are regular employees in laboratory animal facilities and are generally familiar with all the laboratory animal species. They play a significant role in the accreditation of the International Association for the Control of Laboratory Animals (AAALAC-i) or the Korean Excellent Laboratory Animal Facility (KELAF) by the Ministry of Food and Drug Safety. In particular, attending veterinarians play an important role in ensuring the health and a conducive environment for the laboratory animals, which can minimize unexpected variables in well-designed animal studies .This study aimed to design and evaluate a method for improving laboratory dogs and pigs welfare through regular health screening programs conducted by attending veterinarians in an animal research institute. This program is expected to play a pioneering role in laboratory dogs and pigs welfare, which is being strengthened worldwide.Health checkup of the animals was performed according to K-MEDI hub's health checkup procedure until euthanasia at the end of the experiment Fig. . ResearcIn first half of 2019 (1H 2019), the feed amount of dog-#29 increased from 150 g/day to 200 g/day due a body condition score (BCS) of 3/9. This BCS was 4/9 at the medical checkup in the second half of the year. Dog-#39 showed difficulty in feeding due to right upper deciduous canine tooth with bleeding gums. Considering animal welfare, this dog was euthanized with the early termination of experiments, to alleviate pain. In second half of 2019 (2H 2019), the BCSs of dog-#1 and dog-#28 were 6/9. To lose body weight, their feed amounts were decreased from 300 to 250 g. In contrast, since the BCS of dog-#22 was 3/9, the feed amount was increased from 200 to 300 g. Consequently, the BCSs of all the three dogs were 4/9 at the medical checkup in the second half of the year. Although the body weights significantly increased in 2H 2019 (11.51 ± 1.28) and first half of 2020 (1H 2020) (11.72 ± 1.32), compared to those in 1H 2019 (9.31 ± 1.61) (Table 3 cells/uL) decreased to 1.81 × 103 cells/uL, which was within the normal range  for 3 days; on re-examination after 3 weeks, the blood monocyte concentration  and blood biochemistry were performed after aseptic blood collection. In addition, BCSs were measured and specifications were recorded through auscultation, visual inspection, and palpation. BCS is a verified method of scoring animals by physical inspection , 20; theThe BCSs of 3/9 or 6/9 in 4 dogs were normalized to 4/9 or 5/9 by adjusting the feed amount. Optimal body weight is essential for the health and welfare of animals, since BCS ≥ 6/9 (overweight) or BCS ≤ 3/9 (underweight) may be associated with disease or could serve as a predictor of a clinical condition . Dogs wiThe reason BCS was unaffected despite weight gain among dogs in 2H 2019, compared to that in 1H 2019 could be attributed to age-dependent musculoskeletal growth. All the dogs in the present study were beagles, who continue to grow upto 18 months of age . The aveMoreover, ideal BCSs (4/9 or 5/9) were observed among all dogs in 1H 2020, contrary to those among 1H 2019 and 2H 2019 dogs. Therefore, the S.D value of 1H 2020 (4.67 ± 0.71) was smaller than that of 1H 2019 (5.46 ± 0.93) and 2H 2019 (4.53 ± 0.49). Although only one pig in 1H 2020 showed a BCS 2/5, there was no additional clinical symptom. We inferred that this was due to successful regular calibration of BCSs with control of amount of feed and veterinary treatment, including antibiotic use to treat clinical symptoms such as monocytosis and neutrophilia, which can be induced during animal experiments. Therefore, regular health checkups by the AV are imperative to derive reliable results, since because factors causing distress can affect the reproducibility and reliability of animal studies .Despite the commitment to ensure the ideal health of laboratory animals, there are limitations relative to companion animals. First, the dog with bleeding gums due to retained deciduous teeth was euthanized, considering the principle of ‘refinement’ among the 3Rs, since the dog was in severe pain, accompanied by excessive bleeding. Contrary to companion animals, laboratory animals have a wider “humane endpoint”—the scientifically justified point for pain or distress . The humContrary to companion animals, laboratory animals are raised in limited space and are exposed to invasive surgical experiments. It is a methodological challenge for AV to overcome the detrimental environments. Therefore, further studies on regular medical checkups should evaluate the psychological satisfaction of animals according to the provided environmental enrichment and post-operative pain control methods.We introduced highly enhanced animal welfare methods, the history of regular medical checkup established in this study will develop a roadmap for improving welfare and standardization of laboratory dogs and pigs in animal research institute.In conclusion, this study introduced enhanced animal welfare methods; the study design of regular medical checkups established in this study will aid in developing a roadmap for improving the welfare of laboratory dogs and pigs in an animal research institute.K-MEDI hub is a high-tech medical industry cluster created to strategically cater to the global new drug and medical device industries and supports the optimization of new drug candidates, design of medical devices, prototyping, and efficacy evaluation. K-MEDI hub consists of centers for new drug development, medical device development, preclinical research, and clinical drug manufacturing. Among these, the preclinical research center (PRC), which harbors a breeding system for mice, rats, rabbits, dogs, pigs, and monkey is specialized for laboratory animal research such as efficacy/pre-biological stability assessment of new drugs and the performance of new medical devices.The PRC, K-MEDI hub, was certified by the Ministry of Food and Drug Safety in Korea as KELAF in 2016 and fully accredited by the Association for Assessment and Accreditation of Laboratory Animal Care International (AAALAC-i) in 2020. AAALAC-i certification is recognized as an organization for non-clinical studies that encourages the humane treatment of animals in scientific research and ensures optimum animal care and use . These cThe capacities for mice, rats, rabbits, dogs, pigs, and monkey at PRC were 18,000, 1800, 100, 45, 35, and 46 respectively. The K-MEDI hub conducted preclinical evaluations by importing approximately 10,000 mice, 3000 rats, 250 rabbits, 50 dogs, and 40 pigs annually. Moreover, in 2023, they aim to address unmet needs for non-human primate preclinical research by importing roughly 20 non-human primates.Rodent breeding rooms were maintained as specific pathogen free barrier colonies established by the AV and experts in microbiological monitoring. Large animals such as rabbits, dogs, and pigs were reared separately from rodents to prevent cross infection. The IACUC of K-MEDI hub reviews whether exploring alternatives, rationale for the proposed number of animals, minimizing animal discomfort, and humane endpoints are included in the protocol. K-MEDI hub employs one AV who is responsible for animal quarantine, veterinary management, post-approval monitoring, and serving as a member of the Institutional Animal Care and Use Committee (IACUC), with a critical role in ensuring the ethical use of animals in research and maintaining high standards of animal welfare.The post-approval monitoring process of IACUC with the AV semi-annually ensures that the study is being conducted in accordance with the approval and that the researchers protect animal welfare.In this study, to enhance the animal welfare for laboratory animals, PRC, K-MEDI hub itself conducted regular medical check-up programs for laboratory animals, including dogs and pigs, by the AV to ensure the health and welfare of animals and perform appropriate veterinary treatments and environmental management in the facilities.A medical examination was conducted for all the dogs and pigs in breeding. Dogs and pigs were selected as the target species, since blood collection was possible without sacrifice and research involves a relatively long study period, contrary to that in rodents.In 1H 2019, 45 dogs  and 4 pigs  were examined, and in 2H 2019, 24 dogs  and 7 pigs  were examined; 21 dogs  and 14 pigs  were examined in 1H 2020.Blood collected using a sterilized syringe was placed into a serum-separating tube (SST) and an ethylenediaminetetraacetic acid tube and separated from serum in the SST tube.White blood cells (WBC), red blood cells, hemoglobin, hematocrit, mean corpuscular volume, mean corpuscular hemoglobin, mean corpuscular hemoglobin concentration in blood, red blood cell distribution width, platelets, and WBC differential count were analyzed as CBC using a hematology system with an autoslide .The levels of aspartate aminotransferase, albumin, alanine aminotransferase, total bilirubin, triglyceride, blood urea nitrogen, calcium, creatinine, inorganic phosphorus, glucose, sodium, total cholesterol, potassium, total protein, chloride, and C-reactive protein were analyzed as blood biochemistry using a clinical chemistry analyzer .Along with visual inspection, the body weight, auscultation, and palpation were measured along with determining the BCSs of dogs and pigs. Dogs are scored out of 9 and pigs are scored out of 5 , 22. DogStatistical significance was determined using GraphPad Prism 8 . All data are presented as mean ± standard deviation. In case of changes in the body weight of dogs, the normality test was passed and one-way analysis of variance with Tukey's multiple comparisons test was used. P < 0.05 was considered as statistically significant. In case of changes in the BCSs of dogs and the body weights of pigs, normality tests were not passed, and Kruskal–Wallis tests were used.Additional file 1. Supplementary Figure 1. Medical checkup postponement / exemption application form of K-MEDI hub. Supplementary Figure 2. Individual medical record form of K-MEDI hub.Additional file 2. Supplementary Table 1. The results for complete blood cell count in 1H 2019. Supplementary Table 2. The results for blood biochemistry in 1H 2019. Supplementary Table 3. The results for complete blood cell count in 2H 2019. Supplementary Table 4. The results for blood biochemistry in 2H 2019. Supplementary Table 5. The results for complete blood cell count in 1H 2020. Supplementary Table 6. The results for blood biochemistry in 1H 2020.


Estimate the prevalence of chronic pain and its association with symptoms of anxiety, sleep disorders, and aspects of remote work in the context of the COVID-19 pandemic.A cross-sectional and descriptive study conducted with 328 adults engaged in remote work. Data was collected online from February 2021 to January 2022. For pain investigation and evaluations of sleep and anxiety, a structured questionnaire, the Pittsburgh Sleep Quality Index, and the Generalized Anxiety Disorder-7 were used, respectively.The prevalence of chronic pain was 47.9% (CI 95% = 42.5-53.3). Associations were identified between pain and anxiety, sleep disorders, and sitting time (p<0.01).The prevalence of chronic pain in remote work was found to be high, with pain being of moderate intensity and associated with anxiety, sleep disorders, and prolonged sitting time. This led to a reduction in interactions among individuals due to the adoption of remote work, decreased circulation of people in public spaces, closure of businesses, and other measures. However, full-time remote work can have certain drawbacks, including decreased social interaction, limited movement, excessive screen time, and blurred boundaries between professional and personal lives. All these factors can contribute to physical and mental fatigue, stress, anxiety, pain, and exhaustion.Globally, there was a significant push towards the remote work format, also known as teleworking, the emergence of new cases of chronic pain (CP), and the exacerbation of preexisting pain symptoms, 6, especially among those who made drastic changes to their work routines. Pain is known to be multifactorial and can be linked to psychosocial factors. Therefore, during a crisis like the pandemic, the negative effects can be even more pronounced on individuals’ mental, social, and physical well-being.Everyone affected by the COVID-19 pandemic felt its impact due to changes in routine and continuous exposure to physical, psychological, and social stressors. These factors might have led to symptoms of anxiety and sleep disorders, 5, 6, 7 and given the lack of specific research on CP and its correlations with sleep, anxiety, and the nuances of remote work among Brazilians during the pandemic, there was a recognized need to expand knowledge in this unique period. As a result, this study sought to explore the prevalence of CP among remote workers during the COVID-19 pandemic and determine if CP correlated with sleep disorders, anxiety symptoms, and the nature of remote work in that context.Based on the data from studiesTo estimate the prevalence of chronic pain and its association with symptoms of anxiety, sleep disorders, and aspects of remote work during the COVID-19 pandemic in Brazil.The study adhered to both national and international ethical standards and received approval from the Human Research Ethics Committee of the Federal University of São Carlos. All participants provided their consent by clicking on the designated area of the Informed Consent form, which is displayed on the first page of the interview questionnaire.This research is based on a master’s thesis titled “Pain and associated factors in adults working remotely during the COVID-19 pandemic in Brazil.” The study was carried out under the Postgraduate Program in Nursing at the Federal University of São Carlos in 2022.Following the STROBE guidelines, this descriptive, cross-sectional study employed a quantitative methodology. Data was collected online between February 2021 and January 2022 using a structured questionnaire on the Google Forms® platform. Invitations to partake in the research were disseminated via email. Brazilian university graduate programs were asked to forward the invite to faculty and professors, and companies that transitioned to remote work were also targeted. The study was promoted through the Federal University of São Carlos’ institutional network and on the Meta® group’s social media channels, including Facebook, Instagram, and WhatsApp. that focused on the Brazilian populace. For all hypothetical sample sizes, a significance level of 5% was upheld. Variations in the absolute tolerable error ranged from 1 to 8 percentage points (p.p.). Thus, the final sample size for this research fluctuated between 368 to 387 participants, allowing for a 5 p.p. tolerable error.To determine the study’s sample size, calculations were made with the goal of estimating the prevalence of CP in an open-ended population. The prevalence value used in the calculation was sourced from a studyThe inclusion criteria covered adults 18 years and older who were engaged in remote work, be it full-time or part-time, during Brazil’s COVID-19 pandemic. The exclusion criteria ruled out individuals diagnosed with fibromyalgia, rheumatoid arthritis, or cancer, as well as frontline healthcare workers battling COVID-19.To characterize the sample, we designed an online questionnaire to identify sociodemographic and economic profiles, the characteristics of remote work, and lifestyle habits.. Pain intensity was categorized as: 1-4 for mild pain, 5-6 for moderate pain, and 7-10 for severe pain. Pain was defined as chronic if it persisted for a period of 6 months or more from the data collection date.For pain assessment, we structured a questionnaire to ascertain its presence, intensity, location, and duration. To evaluate pain intensity, participants were asked about the average pain they experienced during the pandemic, using an 11-point numerical pain scale ranging from 0 (indicating “no pain”) to 10 (representing the “worst possible pain”) and validated for Brazilian Portuguese by Bertolazi et al.. This instrument, which assesses sleep quality and disturbances, comprises 19 questions. The highest possible score is 21 points, with a higher score indicating poorer sleep quality. A score between 0-4 suggests good sleep quality, 5-10 indicates poor quality, and scores above 10 point to sleep disorders, 13.For sleep evaluation, we used the Pittsburgh Sleep Quality Index (PSQI) questionnaire, developed by Buysse et al., based on the DSM-IV criteria, and was validated by Kroenke et al.. Its translation into Portuguese was done by Pfizer  and subsequently validated in Brazil by Moreno et al.. Comprising 7 items, each is rated on a 4-point scale, ranging from 0  to 3 (nearly every day). The total score can range from 0 to 21, with scores of 10 or higher potentially indicating symptoms of an anxiety disorder.For the assessment of anxiety, we employed the Generalized Anxiety Disorder-7 (GAD-7). This tool gauges the frequency of anxiety signs and symptoms over the past two weeks. It was devised by Spitzer et al., both in simple and multiple forms. A significance level of 5% (p<0.05) was maintained for all analyses.Descriptive statistics were used to characterize the sample. Categorical variables were depicted in both raw and relative frequencies, while continuous variables were represented with means and standard deviations (SD). In the association analyses, we employed the Poisson regression model with robust varianceThe study included 328 participants, with an average age of 32.96 (SD=8) years, ranging between 20 to 61 years. The majority were females (55.2%), and individuals with a completed higher education made up 86.3% of the sample. Socioeconomic and demographic data are detailed in Regarding lifestyle habits, it’s notable that 95.7% of the participants did not smoke, and 62.5% consumed alcohol only socially. Before the pandemic, 61.3% of the sample engaged in physical activity, but this percentage dropped to 52.7% during the pandemic.The prevalence of CP stood at 47.9% . Of the 157 people with CP, 53.50% (n=84) had already felt pain before the pandemic and continued to do so during this period, while 46.49% (n=73) began to experience CP after the pandemic began. The most affected areas of the body were the lumbar spine (62.4%), the cervical spine (47.1%), and the head (42.0%).The average pain intensity during the pandemic was rated at 5.8 (SD=2.0), categorizing it as moderate. Among the participants with CP (n=157), the majority exhibited symptoms of Generalized Anxiety Disorder (GAD) at 53.5%. More than half reported poor sleep quality (59.2%), and 29.9% had sleep disorders. Notably, among people without CP, these percentages were significantly lower: 31.6% with GAD and 11.7% with sleep disorders. The average score on the GAD-7 was 10.9 (SD=5.8), and on the PSQI, it was 8.8 (SD=3.7).Further observation revealed that CP is correlated with sleep disorders (p<0.01). Individuals with CP have a 156% higher prevalence of sleep disorders compared to those without CP .No sociodemographic, economic, or lifestyle characteristics were associated with CP. However, the duration of sitting during remote work correlated with CP. The longer the sitting duration, the higher the prevalence of C P. Individuals who sit for 12 hours a day have a prevalence 3.35 times higher of having C P, and those sitting between 6 and 8 hours have a prevalence 3.28 times higher, compared to those who sit up to 6 hours a day ., 21, 22. However, it should be noted that such studies did not focus on remote workers, which could help explain the elevated rate.The results of the current study revealed a prevalence of 47.9% (95% CI: 42.46%–53.27%) of individuals working remotely with C P, with this pain being of moderate intensity  and associated with symptoms of anxiety, sleep disorders, and prolonged sitting. It can be stated that a 47.9% prevalence of CP during the pandemic is a high figure, especially when considering studies that assessed pain in the general Brazilian population in the years prior to the pandemic. These studies reported rates ranging from 22% to 42% of CP. Another systematic review noted that both the prevalence (95% CI: 0.29 to 0.96) and intensity (95% CI: -2.18 to -0.63) of pain were significantly higher during the pandemic compared to the pre-pandemic period, but it did not specify if the focus was on CP.After the onset of the pandemic, a systematic review of the general population in Brazil (from April to August 2020) showed an average CP prevalence of 45.59%. However, observed CP prevalence in these studies ranged from 23.02% to 76.01%, with the lumbar region (41.96%) being the most affected. Other studies also highlighted significant increases in both pain intensity and prevalence, finding rates between 27.9% and 70.5% of CP in remote workers during the pandemic, 27, 28.Findings similar to our study were identified in a study of adults from Malta (n=388), where the majority were remote workers. 49% of them reported experiencing lumbar spine pain after the pandemic began, whereas 30% had experienced it prior to the pandemic produced different findings: remote work was associated with a reduction in migraine pain intensity during chronic episodes. Psychological symptoms have also been emphasized in the literature due to the shift in people’s routines. For instance, recent studies indicate high levels of anxiety symptoms during the pandemic (47.2% to 56.3%), both in the general population, 31 and among new remote workers, with the latter showing 19.6% to 55.8% anxiety prevalence, 29, 32.In Italy, a study by Curro et al., which found rates between 18.5% and 25.6% of anxiety in migraine sufferers. Bilen and Kucukkepeci found a 66.4% anxiety prevalence in individuals with C P. Nonetheless, there’s a gap in the literature concerning studies linking CP with anxiety symptoms among remote workers during the pandemic. This association between CP and anxiety (p<0.01) was observed in our research, indicating that workers with CP had a 69% higher prevalence ratio compared to those without C P.In our study, 53.5% of those with CP had symptoms of GAD — a high percentage, especially when compared to Curro et al., the authors found that poor sleep quality and increased workload were predictors of anxiety. Curro et al. reported that increased pain intensity was linked to high levels of anxiety and reduced sleep quality and duration. Our study also found an association between pain and sleep disturbances.In the study by Erman et al., 27, 35. Sagat et al. also found that individuals who moved more reported lower pain intensity. It’s worth noting that while sitting posture appears to be a factor in pain, CP is influenced by multiple factors that should be considered.Regarding remote work, our study observed that prolonged sitting is associated with C P, meaning the longer an individual remains seated, the higher their likelihood of developing CP. This is consistent with other studies that found sitting posture to be related to pain intensityThe sample size, initially estimated between 368 and 387 (with a tolerable error of 5 percentage points), was not achieved. Therefore, an approximate tolerable error of 5.4 percentage points is admitted. According to stratification by regions of Brazil, the sample size was only achieved in the Central-West (67) and Southeast (196) regions. This represents a significant limitation of the study.The lack of investigation regarding the intensity of CP before the pandemic is also a limitation. This information could have been collected for a better understanding of the situation, although it would be a subjective data relying on the recollection of pain intensity perception. However, it would be useful for comparative purposes.Finally, the remote mode of data collection can also be considered a limitation, as it doesn’t allow for a diverse sample and limits potential participants’ access due to outreach issues .This study is relevant because it provides important information about remote work, a work arrangement that became more common during the pandemic. This work format has both positive and negative aspects, as identified during this period. Therefore, it is important to pay attention to the physical and mental health of remote workers to understand how changes in their routines might lead to pain and other comorbidities. This research can help develop strategies and interventions in both the public and private sectors to minimize the negative effects of remote work.This research provides valuable insights for health professionals and pain specialists by providing an overview of chronic pain, anxiety symptoms, and sleep disorders in remote workers in Brazil. Companies and health professionals must work together to establish preventive and therapeutic measures for these symptoms in this group, as many companies are likely to continue to adopt remote work.We identified CP prevalence in approximately half of the Brazilian remote workers sample during the COVID-19 pandemic. The pain intensity was moderate among them, with the lumbar spine being the most affected region. Moreover, about half of those reporting CP manifested this symptom during the pandemic. Most showed GAD symptoms, and nearly a third of those with CP experienced sleep disorders. In contrast, in those without chronic pain, these rates were significantly lower. CP pain was associated with GAD symptoms, sleep disorders, and prolonged sitting.0034-7167-reben-76-s1-e20230012-suppl01Click here for additional data file.