MethodsX

Oxidative stress results from excessive accumulation of reactive oxygen species (ROS) and plays a central role in numerous physiological and pathological processes. Accurate quantification of antioxidant enzyme activities is therefore essential in redox biology research. However, data analysis for commonly used assays, such as superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPx), is frequently performed using spreadsheets or manual calculations, which are time-consuming and prone to error. Here, we present Redoxyme, a free, open-source, Python-based graphical user interface designed to standardize and automate the calculation of antioxidant enzyme activities. The software integrates protein normalization, enzyme-specific calculation routines, data visualization, and Excel export within an intuitive interface that does not require programming expertise. Redoxyme was validated using experimental data obtained from animal tissues (rats and mice), demonstrating excellent agreement with manual calculations and established analytical methods. Redoxyme provides a practical solution for improving reproducibility and efficiency in antioxidant enzyme activity analysis. The software is currently distributed as a standalone executable for Windows (locally installed), and an interactive web-based calculator implemented in Streamlit, enabling direct use without local installation. The source code and version-controlled development history are openly accessible via GitHub, promoting transparency, reproducibility, community-driven improvements, and can, in principle, be adapted for other operating systems. Graphical Abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=63 SRC="FIGDIR/small/703993v2_ufig1.gif" ALT="Figure 1"> View larger version (10K): org.highwire.dtl.DTLVardef@120cc68org.highwire.dtl.DTLVardef@4be246org.highwire.dtl.DTLVardef@1f47134org.highwire.dtl.DTLVardef@1341100_HPS_FORMAT_FIGEXP M_FIG C_FIG

Holographic imaging in microscopy enables label-free quantitative information of biological specimens and has found applications across a wide range of biomedical studies, from cell morphology to particle dynamics; yet its widespread adoption is often limited by the lack of accessible and standardized analysis software. We present HoloBio, an open-source, Python-based graphical user interface developed to address this issue. This software offers two primary operational modes: a Real-Time mode that enables live processing of holograms at video frame rates, and an Offline mode designed for post-processing previously recorded holograms. HoloBio is compatible with holograms recorded using both lens-based and lensless systems, supporting off-axis architectures in telecentric and non-telecentric configurations, as well as slightly off-axis and in-line optical setups. The software incorporates tools for cell tracking, phase profiling, thickness estimation, and morphological analysis, including cell counting and object area quantification. HoloBio is designed to be accessible for users without coding expertise, offering a reproducible, high-throughput environment tailored for researchers in biology, biophotonics, and biomedical imaging.

Investigating single-cell dynamics and morphology in tissues and embryos requires highly accurate quantitative analysis of microscopy images. Despite significant advances in the field of bioimage analysis, even the most sophisticated segmentation and tracking algorithms inevitably produce errors (e.g. : over segmentation, missing objects, miss-connected objects). Although error rate may be small, their propagation throughout a time-lapse sequence has catastrophic effects on the accuracy of tracking and extraction of single cell parameters. Extracting single cell temporal information in the context of tissue/embryo requires thus expert curation to identify and correct segmentation errors. In the movies commonly used in developmental biology and stem cell research, both the number of imaged cells and the duration of recording are large, making this manual correction task extremely time-consuming. This has now become a major bottleneck in the fields of development, stem cell biology and bioimage analysis. We present here EpiCure (Epithelial Curation), a versatile tool designed to streamline and accelerate manual curation of segmentation and tracking in 2D movies of large epithelial tissues. EpiCure uses temporal information and morphometric parameters to automatically identify segmentation and tracking errors and provides user-friendly tools to correct them. It focuses on ergonomics and offers several visualization options to help navigating in movies of tissue covering a large number of cells, speeding up the detection of errors and their curation. EpiCure is highly interoperable and supports input from a wide range of segmentation tools. It also includes multiple export filters, enabling seamless integration with downstream analysis pipelines. In this paper, using movies from several animal models, we highlight the importance of curating cell segmentation and tracking for accurate downstream analysis, and demonstrate how EpiCure helps the curation process for extracting accurate single cell dynamics and cellular events detection, making it faster and amenable on large dataset.

AimsThe development of a method for isolating mitochondria from a specific cell type within a given tissue, while preserving their structural and functional integrity to the greatest possible extent, remains an ongoing challenge. The aim of this study was to establish a protocol for the isolation of mitochondria from rodent cardiomyocytes, characterized by minimal contamination with other cell types and a high yield of mitochondrial fractions originating from distinct subcellular regions of cardiomyocytes. Methods and resultsIn the present study, cardiomyocytes from guinea pig and rat hearts were isolated using a standard enzymatic digestion protocol in a Langendorff heart perfusion system. Traditionally, the isolation of organelles, including mitochondria, from whole cardiac tissue as well as from cardiomyocytes has relied primarily on mechanical tissue homogenization These conventional approaches involve the localized application of high pressure to cells, which may potentially damage delicate organelles, particularly mitochondria. Moreover, such homogenization preferentially releases mitochondria located in the subsarcolemmal region of cardiomyocytes rather than representing the entire mitochondrial population. In our study, we employed an alternative approach based on the gentle mechanical disruption of cardiomyocytes by passing the cell suspension through selected cell strainers using a cell scraper. This strategy facilitated mild disruption of cellular structures, significantly increasing the yield of mitochondria released from interfibrillar regions while preserving mitochondrial functionality. Moreover, this method decrease probability of sample contamination with mitochondria from other cells, based on cell size differences. The effectiveness of this method was confirmed by transmission electron microscopy, and high-resolution respirometry, which revealed no evidence of outer mitochondrial membrane damage, as indicated by the lack of response to the addition of exogenous cytochrome c to the incubation chamber. Moreover, mitochondrial oxygen consumption increased by 7.39 {+/-} 1.25-fold following the addition of 100 {micro}M ADP, reflecting efficient ADP-stimulated respiration. Furthermore, fluorescence measurements were performed. to assess changes in the mitochondrial inner membrane potential ({Delta}{Psi}). The isolated mitochondria were also suitable for electrophysiological studies using the single-channel patch-clamp technique. Additionally, mitochondria isolated using the protocol developed in our laboratory exhibited a high capacity for transplantation into H9c2 cells. ConclusionIn summary, our mitochondrial isolation method is rapid, efficient, and yields functionally competent mitochondria. These preparations are suitable for a wide range of downstream applications, including patch-clamp electrophysiology, analyses of oxygen consumption under various pharmacological conditions, as well as mitochondrial transplantation. Graphical abstract O_FIG O_LINKSMALLFIG WIDTH=162 HEIGHT=200 SRC="FIGDIR/small/716092v1_ufig1.gif" ALT="Figure 1"> View larger version (85K): org.highwire.dtl.DTLVardef@613495org.highwire.dtl.DTLVardef@1c34338org.highwire.dtl.DTLVardef@722900org.highwire.dtl.DTLVardef@e1f7a6_HPS_FORMAT_FIGEXP M_FIG C_FIG

Objectives: To determine the impact of discharge home on physical activity and sedentary behaviour following orthopaedic trauma. Design: Observational study. Setting: Acute hospital. Participants: Between October 2022 and January 2024, 31 adult orthopaedic trauma patients were recruited during hospital admission. Participants had either an isolated hip fracture or multi-trauma (i.e., a lower limb fracture, with an upper limb and/or spinal fracture). Interventions: Participants wore two activity monitors (activPAL3 and ActiGraphGT3x) during the final days of an acute hospital admission and the first five days at home. An interrupted time series analysis evaluated changes physical activity variables during the hospital to home transition. Participants were analysed individually using mixed-effects linear regression allowing the intercept to vary by participant. Main outcome measures: Primary outcome was daily steps; secondary outcomes included sedentary time and other activity measures. Results: Daily steps (mean +- SD) were higher at home (4552.4 +- 2639.5) compared to hospital (2597.8 +- 1450.8). Modelled results indicated a 27% increase in daily steps following hospital discharge (exp(beta946;): 1.27, 95% CI: 1.01,1.59, p=0.039) and a sustained improvement at home. No significant differences were observed between hip fracture and multi-trauma participants. Conclusion: Participants recovering from orthopaedic trauma showed a significant increase in daily step count upon discharge home from hospital, highlighting the positive impact of the home environment on activity levels. Further research is warranted to assess the effectiveness of interventions to improve activity levels in hospital (e.g., early intensive therapy) and at home (e.g., immediate home-based physiotherapy) in individuals following orthopaedic trauma.

Precision-cut lung slices (PCLS) have emerged as a powerful tool for studying the biology of viable human lung tissue. However, the presence of agarose impurities compromises RNA yield and integrity during the extraction process. We tested whether using an alternative Plant kit RNA extraction method to wash agarose impurities or pre-dissolving agarose from PCLS implementing a dissolving buffer for routine RNA isolation in gel-electrophoresis would improve RNA quantity, quality, and integrity. Our results show that RNA quantity and integrity are highly compromised when using a conventional method of RNA extraction. The plant kit and dissolution of agarose increased the RNA quantity to 0.42{+/-}0.11 and 0.65{+/-}0.17 {micro}g/PCLS (measured by the Qubit) and integrity number to 6.60{+/-}0.59 and 9.13{+/-}0.39 (measured by the Bioanalyzer), respectively. The presence of impurities in conventional and Plant kit extractions misled to an overestimation of the RNA quantity and quality using the NanoDrop. The Plant kit and agarose dissolution showed a significant transcript integrity increase in GUSB (p<0.0001) and COL1A1 (p<0.05) expression, validating these methods over conventional extraction. We encourage laboratories applying PCLS experimentation to implement alternative methods to remove agarose impurities during RNA extraction, as well as to rely on sensitive quantitative techniques, such as the Qubit and Bioanalyzer, for RNA quantification and integrity measurements.

Quality and price of fetal bovine serum (FBS) are traditionally determined by geographical origin and parameters listed in the Certificate of Analysis (CoA). Despite its central role in cell culture, selecting suitable FBS batches remains costly and labor-intensive due to substantial batch-to-batch variation. We propose a molecular assessment strategy based on transcriptomic and cytokine profiling of cells cultured in different FBS batches to evaluate performance more reliably. Analysis of differential gene expression in three cell lines - MRC-5, Jurkat, and THP-1 - enables batch grouping and reveals pathway-specific effects, with immune-related pathways showing the most pronounced variability. Although CoA parameters can stratify batches by origin, they do not consistently correlate with cytokine secretion or gene expression across cell lines. These findings demonstrate that geographical origin is an inadequate predictor of functional FBS performance and that molecular profiling provides a more robust and informative assessment.

BackgroundQuantitative myocardial blood flow (MBF) and myocardial flow reserve (MFR) provide incremental diagnostic and prognostic value in cardiac PET, but their widespread use is limited by the technical demands of dynamic imaging protocols. We evaluated the feasibility of using artificial intelligence (AI) to predict MBF and MFR from static and gated PET images, without the need for dynamic acquisition. MethodsA machine learning (XGBoost) model was trained on 82Rb PET multi-center dataset using static perfusion imaging, injected dose, hemodynamic measures, clinical data and CT-derived features (including body composition) from the hybrid CT attenuation scan. Model performance was evaluated externally in an independent cohort. ResultsIn total, 10,566 (derivation-cohort) and 7,842 (external-cohort) patients were included in this multi-center study. On the external-cohort, AI approach achieved an Area under the curve (AUC) of 0.92 (0.92-0.93) for abnormal stress MBF and 0.91 (0.90-0.92) for abnormal MFR; Intra-class correlation (ICC) 0.80 (0.78-0.82) and 0.78 (0.76-0.79), respectively. AI MFR closely mirrored the prognostic performance of measured MFR, showing nearly identical Kaplan-Meier risk stratification (both p<0.0001) and maintaining strong, and independently significant associations with all-cause mortality (HR 3.4 [2.8-4.2] vs. 4.6 [3.6-5.8]; both p<0.001), and demonstrated similar added value to perfusion for mortality prediction. ConclusionAI-predicted virtual stress MBF and MFR assessment using static and gated PET data is feasible and generalizable across cohorts. By removing the dependency on dynamic acquisitions, this approach has the potential to broaden the clinical adoption of flow quantification. O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=97 SRC="FIGDIR/small/26345376v1_ufig1.gif" ALT="Figure 1"> View larger version (27K): org.highwire.dtl.DTLVardef@ec2522org.highwire.dtl.DTLVardef@17a04aforg.highwire.dtl.DTLVardef@1c99db7org.highwire.dtl.DTLVardef@1918c8f_HPS_FORMAT_FIGEXP M_FIG STRUCTURED GRAPHICAL ABSTRACT PET: Positron Emission Tomography, CT: Computed Tomography, MFR: Myocardial Flow Reserve C_FIG Key Question: Can machine learning models trained on dynamic PET datasets accurately predict regional stress myocardial blood flow (MBF) and myocardial flow reserve (MFR) from static image features, physiological parameters, and CT-based anatomical measures? Key Finding: Artificial intelligence can accurately estimate MBF and MFR from non-dynamic PET data, with strong agreement to reference standards. Take-home Message: By eliminating reliance on dynamic PET acquisitions, machine-learning has the potential to broaden clinical adoption of quantitative flow assessment.

ContextAs global urbanization intensifies, Urban Green Spaces (UGS) are pivotal for biodiversity conservation and climate change mitigation. However, comparative assessments of UGS spatial configuration and connectivity across diverse urban landscapes remain limited. ObjectivesThis study aims to assess the spatial arrangement and connectivity of UGS across 28 European capital cities. Additionally, we evaluate how Network Science metrics derived from Graph Theory can complement traditional landscape ecology metrics to provide a more comprehensive understanding of UGS at a large scale. MethodsWe developed a European Urban Vegetation Map using Earth observation data to classify UGS at 10m resolution across the selected capitals. We then analyzed UGS connectivity for each city utilizing 40 traditional landscape metrics and a Graph-Theory-based approach. ResultsWhile traditional landscape metrics effectively quantified fragmentation, they often remain strongly correlated with total vegetation abundance. In contrast, Network Science metrics provided specific insights into UGS functional connectivity, distinguishing the quality of ecological links beyond spatial proximity. This integration allowed us to cluster European capitals into three distinct typologies: unconnected compact cities, large metropolises with complex peri-urban dynamics, and high-connectivity cities with robust networks. These findings demonstrate that graph-based indices effectively complement traditional metrics, highlighting that relying solely on green space percentage is insufficient for assessing the ecological resilience of urban environments. ConclusionsThese results underscore the relevance of Earth observation-based UGS assessment and demonstrate that graph-based landscape connectivity analysis outperforms simple abundance metrics. Therefore, effective assessment requires integrating structural metrics with graph-based connectivity to support resilient urban biodiversity.

AimAs cities densify and expand, the careful planning and design of green spaces are essential for supporting urban biodiversity. Here, we evaluate the relative contribution of habitat patches of varying size, quality, and connectivity to urban biodiversity and assess environmental factors driving differences in species richness and community composition. LocationZurich, Switzerland. Time period: 2008-2018.Major taxa studied: Invertebrates, vertebrates, and trees. MethodsWe analyzed species occupancy data from 452 habitat patches. We quantified alpha, beta, and gamma diversity, assessed species-area relationships, and applied generalized dissimilarity modelling to test the role of patch area, connectivity, and habitat quality--proxied through environmental variables, including vegetation complexity, water presence, and forest isolation--in shaping community composition. ResultsAlpha diversity increased significantly with area, although small patches (usually < 5 ha) disproportionately contributed to beta diversity. Per unit area, groups of small patches yielded higher gamma diversity than equivalent areas of large patches, particularly for trees and invertebrates. Community composition was strongly influenced by patch area, with effects mediated by vegetation complexity, water, and isolation, with responses differing among taxa. Main conclusionsSmall habitat patches play a critical role in enhancing overall urban biodiversity. They increase species richness through cumulative area effects and promote community turnover (mediated by environmental heterogeneity). Maintaining networks of small patches alongside large green spaces is therefore key to conserving biodiversity in urban landscapes.

Exercise training is a cornerstone of Cardiovascular Rehabilitation (CR) and, as of now, moderate-to-vigorous continuous exercise training (MICT) is the standard. New exercise modalities in the context of CR are constantly being explored to improve patient outcomes. These Canadian Association of Cardiovascular Prevention and Rehabilitation (CACPR) exercise training recommendations provide a synthesis of evidence-informed recommendations from existing documents, including recommendations around High-Intensity Interval training (HIIT). CACPR created a pan-Canadian Exercise Working Group with various knowledge users (e.g., kinesiologists/exercise physiologists, physiotherapists, cardiologists, and patients) with expertise in CR-based exercise, who developed knowledge gap questions related to exercise training based on a literature review and synthesis of all available recommendations. An independent evidence-synthesis team performed a rapid review and meta-analyses to address the questions. The working group used this data to develop relevant recommendations. The final guidelines include 12 recommendations for CR exercise, including nine from previous documents and three new recommendations based on HIIT. The previous recommendations address exercise assessments and prescriptions for CR for various patient profiles. The new recommendations suggest that HIIT can be used to improve exercise capacity in patients with coronary artery disease (CAD), heart failure (HF) or atrial fibrillation. They also state that HIIT is superior to MICT in patients with CAD, that patients with HF should be considered for either HIIT or MICT and that any HIIT interval duration can be used as part of CR. Overall, these recommendations provide guidance for exercise in Canadian CR programs.

O_LIBiodiverse vegetation that supports high rates of ecosystem functions can inherently express a messy appearance due to high numbers of local native plant species and their spatial distribution connected to niche complementarity. This messiness is assumed to lower peoples appreciation for vegetation in urban contexts. Since such vegetation and a positive relationship between people and biodiversity could contribute to mitigating biodiversity loss, this assumed low public appreciation warrants investigation. C_LIO_LIWe designed and constructed biodiverse flowerbeds using only local native plants, and with the intention to enhance planting productivity, resistance and resilience. To investigate the influence of messiness, we created flowerbeds in four high levels of species richness (8,12,16,20) shown to be relevant for ecosystem functioning, and three levels of order (no, semi, full). We tested public appreciation for the flowerbeds using a self-guided, on-site survey. C_LIO_LIWe found a positive mean rating for all flowerbeds, but no effect of species richness on the ratings. Increased order, however, had a strong negative effect: The odds of a fully ordered flowerbed receiving a negative rating were 88% higher than of a flowerbed with no order. Increasing designed order was correlated with decreasing plant biomass in the flowerbeds. C_LIO_LIThese findings challenge the assumption that the appearance of biodiverse plantings is too messy for public appreciation in urban contexts. Specifically, we demonstrate that introducing order and reducing messiness can compromise aesthetic appreciation for biodiverse vegetation, potentially by compromising productivity as indicated by lower biomass production in ordered plantings. C_LI Synthesis and applicationOur study shows that biodiverse vegetation can be appreciated in urban contexts. Flowerbeds can effectively serve both people as ornamentation and biodiversity as habitat when they are designed based on ecological principles Research highlightsO_LIWe designed and realized flowerbeds based on ecological principles. C_LIO_LIAll plantings received positive average ratings. C_LIO_LIIncreasing species richness in the flowerbeds did not affect participants aesthetic appreciation. C_LIO_LIIncreasing order in the design of the flowerbeds strongly lowered participants aesthetic appreciation. C_LIO_LIIncreasing order in the design was correlated to lower biomass productivity and more bare soil in the flowerbeds. C_LI

Aims: Explainable advanced electrocardiography (A-ECG) can be used to estimate heart age from the standard 12-lead ECG. A-ECG heart age gap (HAG) represents the difference between A-ECG heart age and chronological age. Increased A-ECG HAG is associated with cardiovascular outcomes and can be used to communicate risk. The aim was to investigate whether A-ECG heart age demonstrates acceptable within- and between-session reproducibility. Methods: Healthy adults (n=42, age 23+/-4 years, 52% male) attended up to two sessions ~14 days apart, with 36 participants completing both sessions. During each session, five standard resting 12-lead ECGs were obtained while lying in the supine position with unchanged electrode positions. A-ECG heart age was extracted using dedicated software. Within-session reproducibility was assessed using all five recorded ECGs with coefficient of variation (CV) and a two-way random effects intraclass correlation coefficient (ICC). Between-session reproducibility was assessed using the first recorded ECG of each session with a paired t-test, CV and ICC. A further analysis assessed the reproducibility of the parameters used in the A-ECG heart age regression model. Results: A-ECG heart age showed excellent within-session reproducibility in session one and two (both CV 5.8%, ICC 0.99). A-ECG heart age was slightly lower in session one than two (24.0+/-7.5 vs. 25.5+/-7.8 years, p=0.04) and showed good between-session reproducibility (CV 8.3%, ICC 0.84). All but one parameter used to estimate A-ECG heart age showed acceptable within- and between-session reproducibility (CV<10%). Conclusion: A-ECG heart age demonstrates excellent within-session reproducibility and good between-session reproducibility in healthy young adults.

ObjectiveGeneration and testing of IGF1 reference materials (RM), suitable for the harmonization of immunoassay (IA) and LC-MS/MS methods for the IGF1 determination in blood. In addition, establishment of age related reference intervals for men and women. MethodsIn a split sample study of 42 patients, and 30 healthy volunteers we tested the commutability of four RMs for IGF1, using four commercial IAs and an LC-MS/MS method. A new set of age dependent reference intervals was established using Lifelines biobank samples, based on the IGF1 LC-MS/MS method. ResultsThe four RMs were found to be commutable, except the RM with the lowest concentration measured with the Siemens Immulite method. The value assignment of the RMs was based on the IGF1 LC-MS/MS method, which was calibrated against WHO international standard 02/254. LC-MS/MS results were on average about 0 to 60% lower than those of the immunoassays. Combining the recalculated IGF1 results in patient samples from a former study with the data from healthy volunteers in this study, showed a reduction in the variation of the data points (standard error of estimate) of 42% and 62% respectively. ConclusionCommutable RMs for IGF1 can be made from serum of healthy blood donors. However, it remains necessary to test the commutability of these RMs in IAs that were not included in this study. By harmonizing methods using the four RMs, the same age-related reference intervals can be used.

Global healthcare is targeting patient-centred care, as it leads to better health outcomes and higher level of patient satisfaction. Patient-centred communication, is an important part of patient-centred care because it focuses on involving patients in their care. Recent surveys both nationally and globally have shown that patients are not involved enough in their own healthcare decisions. This problem is especially common among the elderly with chronic conditions. This study aimed to describe patient-healthcare professional interactions, expectations, and satisfaction in physiotherapy within an understudied context, thereby providing important, specific data on ICE dynamics and satisfaction in the specific setting. Cross-sectional study of participants in scheduled consultations was conducted. Two government physiotherapy centres, seven private physiotherapy centres and two trust centres with physiotherapy facilities in Gujarat, India. 232 patients (from various public and private physiotherapy clinics) participated in the study. Patients' ideas, concerns, expectations (ICE) and satisfaction were explored. Almost 88% of patients reported their thoughts and explanations about their symptoms during the consultation. Most patients described not having any concerns about the diagnosis/treatment, and more than two-third of patients consulting PTs expected explanation for their symptoms. Almost 90% patients were satisfied with the consultation. The study revealed that while most patients conveyed their thoughts during consultations, very few expressed their concerns. Overall, patients were satisfied with their consultations.

Recent progress in mathematical kinship modelling has allowed one to predict the probable numbers of kin for a typical population member. In the models, kin may be structured by age and sex, both in static or time-variant demographies. Knowing the probable numbers of kin in different stages - such as parity, health status, or geographic location - however, remains an open challenge in Kinship Demography. Knowing how population structure delimits kin to distinct stages is an advance - for instance, the probability of having one sister at home and one sister away has different social implications from the probability of having two sisters. We present a novel analytical framework, grounded in branching process theory, that provides kin-number distributions jointly structured by age and stage. Using recursive compositions of probability generating functions (PGFs), we derive the joint age, stage, and age x stage kin-number distributions. All marginal distributions over either dimension naturally emerge. Simple extensions of the PGF approach additionally yield: the joint distribution of an individuals own stage and their kins stage; the probable numbers of kin deaths, both in total and by generation number; and the probabilities of being kinless and/or orphaned. We demonstrate the framework through novel results in an application using UK parity-specific fertility and mortality data. HighlightsO_LIA new method calculates probability generating functions for the number of kin structured by age and stage C_LIO_LIThe model allows predicting the probable numbers of kin organised by age and stage C_LIO_LIRecursive nesting of probability generating functions in branching processes is used C_LIO_LIAn application is presented highlighting the novel results C_LI

IntroductionSupervised statistical learning for cell-level segmentation and morphometry in optical microscopy is limited less by algorithmic capacity than by the scarcity of reliable, expert-validated ground truth. In comparative neuroscience and quantitative histology, where classical stains such as Nissls method remain the primary means to study cellular morphology, this bottleneck is acute: manual annotation is expensive, subject to individual bias, and rarely performed at the scale or consistency that computational approaches demand. No existing platform integrates a stain-specific bioimage segmentation protocol, a structured multi-annotator workflow, and consensus-based quality control into a single pipeline from image ingestion to machine-readable training data. MethodsWe present Anatolution, an open-source, web-based platform designed to address the gap of quality annotations at https://anatolution.herokuapp.com/public-tool/. Anatolution organizes microscopy images, including 2D arrays or 3D volumes, into project workspaces where multiple annotators independently label cellular structures against a shared computer vision catalogue. This design enables systematic inter-rater and intra-rater reliability assessment, with consensus derived from agreement across annotators rather than from any single experts judgment. The platform enables the export of aggregated labels or annotation datasets for downstream statistical learning methods. We describe the systems architecture, its Nissl-specific segmentation pipeline, the consensus annotation workflow, and validation of inter-rater reliability. ConclusionAcross 20+ histological annotation containers annotated by up to 15 independent raters, consensus boundary agreement increased monotonically with annotator count, reaching a median Dice of 0.79 against the full-rater reference at seven annotators, with top-tier containers achieving leave-one-out ceiling values of 0.621-0.769 for cell-body segmentation. The segmentation pipeline provided effective spatial anchoring, with 88% of consensus-annotated polygons containing at least one algorithmically detected seed. Anatolution provides open-source infrastructure for producing consensus-validated training data from classical histological preparations, addressing the primary bottleneck limiting supervised learning for cell-level morphometry.

The protein sample preparation methods for shotgun proteomics are nowadays well-established unlike the ones for whole protein analysis. The goal of my work has been to create a simple methodology which provides a single uncomplicated sample preparation tool for these two fields. Nowadays the bulk of proteomics work is done using detergents for protein solubilization. The presented concept, which is based on unspecific adsorption of protein molecules on wide pore materials, allows for protein capture and clean-up from solutions of the most commonly used sodium dodecyl sulfate detergent. It could also be applied to proteins in detergent-free solutions. After the capture and clean-up, proteins could be either cleaved for the downstream peptide analysis or eluted for the whole protein analysis. If required, the eluted whole proteins could be recaptured and cleaved into peptides. Depending on the experimental goals, the sample preparation device could be fitted with embedded proteolytic enzymes to simplify routine sample processing and/or reversed phase media for the downstream peptide or protein separation.

IntroductionIn Canada, older adults face barriers to access healthcare, which includes falls prevention. An effective option to improve access is telehealth, which may be used for falls risk screening remotely. However, reliability and validity of remote functional mobility assessment have yet to be established within a francophone older adult context. This study aimed to assess the reliability and validity of the Timed Up and Go and the Five Times Sit to Stand when administered remotely and determine whether remote assessments could serve as a valid and reliable alternative to in-person fall risk screening for francophone older adults. MethodsThe TUG and FTSTS were conducted remotely by physiotherapists and in-person by individuals with varied healthcare backgrounds. Inter-rater reliability was examined between two remote assessors and between one remote compared to one in-person assessor. Concurrent validity was examined between in-person and simultaneous remote assessments. ResultsSixty-seven older adults completed assessments either with an in-person and a remote assessor or two remote assessors. Excellent inter-rater reliability was documented for both the TUG and FTSTS remotely and in-person. Concurrent validity was also excellent, with complete agreement between remote and in-person assessors for correctly identifying participants at risk of falls. DiscussionRemote mobility assessments can be conducted safely and with excellent reliability, providing an effective alternative to in-person falls screening in a preferred language. Remote assessments can be used in the context of falls prevention programs to improve access and health equity for older individuals who face mobility, geographic or linguistic barriers.

BackgroundReactive balance training using repeated perturbations may reduce falls, however, training methods are not easily replicated or translatable to clinical settings. This study aimed to examine the effects of a novel reactive balance training program on balance recovery from laboratory induced trips and slips and fall risk factors in older people using simple and low-cost equipment. MethodsWe conducted a randomised controlled trial involving 88 older people. An intervention group (n = 43) received the ReacStep program which involved tether-release reactive stepping and intentional slips once a week for 6 weeks. Both the intervention and control (n = 45) groups received home-based strength training for 8 weeks. Blinded staff assessed reactive balance (laboratory induced falls), physical functions at baseline (week 1) and post intervention (week 8). Weekly SMS surveys ascertained falls in daily life over 12 months. ResultsBoth groups were comparable in demographics, with a mean age of 72 years (SD = 5.6). Adherence to ReacStep sessions was high (90%). There were no significant differences between groups in laboratory-assessed reactive balance falls at post-test or daily-life falls over one year (P =.19). However, at post-test, the intervention group demonstrated significant improvements in usual gait speed, maximum step length, and choice stepping reaction time compared to controls (P <.05). ConclusionsThe ReacStep program demonstrated excellent adherence, was well tolerated, and improved gait parameters required for balance recovery following postural perturbations in older people. Nevertheless, it appears this program is not sufficient to improve reactive balance against unexpected trips and slips. Key pointsO_LIThe ReacStep program is acceptable, demonstrates excellent adherence and improves gait measures in older people, potentially reducing fall risk. C_LIO_LIThe generalisability against unexpected trips, and slips, and falls in daily life may be limited. C_LIO_LIFuture research should explore more ecological perturbations while maintaining its accessibility and acceptability. C_LI