MethodsX

Motion artifacts present a major challenge for intravital imaging of tissues undergoing physiological movement or mechanical loading. Blurring or artificial changes in image intensity due to shifting on the z-axis reduce data reproducibility and reliability. Existing post-acquisition approaches can partially compensate for motion, but they increase experimental complexity and are not well suited for use in mechanically loaded bone. Therefore, we developed a novel method for real-time correction of axial motion during mechanical loading of bone by synchronizing the movement of the objective to the actuator. Synchronization was achieved by linking the position of the actuator piezo motor to the objective piezo motor with a user refined reduction via potentiometer. Applying axial motion correction effectively removed artificial changes in fluorescent intensity in a static fluorescent marker up to 3000{varepsilon} in bone as measured by similarity and average intensity before and during loading. This improvement was reflected in the improved accuracy in capture of a dynamic fluorescent calcium indicator (GCaMP6f) in osteocytes. Our system provides a user-friendly, robust framework that can be easily adapted to other mechanically loaded tissues, improving data collection and expanding the utility of two-photon imaging across a variety of biological applications.

Shoulder impingement syndrome (SIS), including subacromial impingement and rotator cuff tendinitis, is commonly caused by repetitive swimming movements and associated shoulder joint dysfunction. Despite numerous available treatment options, no consensus exists on the most effective treatment option. Therefore, this systematic review and network meta-analysis aimed to investigate treatment methods for SIS in swimmers. Using a frequentist framework and Cochrane PICOS principles, we compared SIS treatments, constructed network evidence diagrams, and assessed heterogeneity. A total of 45 studies were included in the qualitative synthesis, and 42 contributed to the network meta-analysis, comprising 1752 participants, 9 treatment categories, and outcome measures. For pain outcomes, some adjunctive interventions combined with exercise showed favorable ranking probabilities, although several estimates were accompanied by wide confidence intervals. For shoulder range-of-motion outcomes, taping, acupuncture, manual therapy, and sport-specific training showed favorable effects in selected comparisons, particularly for external and internal rotation. According to surface under the cumulative ranking curve (SUCRA) rankings, exercise combined with medium-frequency therapy ranked highly for pain reduction, whereas exercise combined with acupuncture or extracorporeal shock wave therapy ranked highly for shoulder flexion. Exercise combined with taping ranked highly for external rotation, and exercise combined with manual therapy ranked highly for internal rotation. However, the interpretation of ranking results should remain cautious because uncertainty and inconsistency were present in some comparisons. Exercise-based rehabilitation appears to remain central to the management of SIS in swimmers. Several adjunctive interventions showed favorable findings for selected outcomes, especially pain relief and shoulder rotational function. However, the available evidence was affected by heterogeneity, inconsistency, and imprecision across some treatment comparisons. More rigorously designed swimmer-specific randomized controlled trials are needed before firm treatment hierarchies can be established. Trial registration: The protocol for this systematic review is registered with PROSPERO (www.crd.york.ac.uk/PROSPERO; registration number: CRD42024498851). The first submission of PROSPERO was on January 15, 2024, and it was revised and updated on March 25, 2026.

Asymptotic standard errors for the parameters of a nonlinear mixed-effects model fitted by first-order conditional estimation (FOCE) or FOCE with interaction (FOCEI) require the observed (Fisher) information -- the negative second derivative of the population objective at the optimum. The gradient of this objective can be computed exactly from sensitivity equations, but the observed information is conventionally still formed by finite differencing, which is less accurate and step-size dependent. Our objectives are to (i) derive the FOCE and FOCEI observed information in closed form within the same sensitivity-equation framework, and (ii) quantify the precision this recovers. Writing the objective as a data term plus the log-determinant of the first-order inner Hessian, the population Hessian splits so that the data term reuses the second-order sensitivities already needed for the gradient, whereas the log-determinant term requires third-order sensitivity equations -- confining the third-order dependence to a single term, where it enters in exactly two places. A finite-difference error analysis shows the differenced Hessian attains an accuracy no better than the square root of the objectives evaluation accuracy, whereas the analytic form is limited only by the sensitivity and differential-equation solutions, with no step size to tune. We illustrate this on a one-compartment oral model with first-order absorption fitted to warfarin data, where the differenced standard error is usable only over a narrow band of step sizes while the analytic value carries none. Implemented in the open-source R package nlmixr2, the method makes exact, reproducible standard errors routine, supporting more dependable confidence intervals, identifiability assessment, and uncertainty propagation.

To explore adapting qPCR systems for end-point nucleic acid quantification using dyes such as SYTO-9, we quantified serial dilutions of DNA and RNA standards in the range of 0.75 - 200 ng/{micro}l on 384-well qPCR devices. SYTO-9 fluorescence was successfully measured using standard SYBR Green settings. Blank-subtracted relative SYTO-9 signal showed a logarithmic dependence on DNA/RNA concentration (R2 > 0.95). Measurements were highly stable with different incubation times, temperatures of up to 95{degrees}C, and photobleaching. The described approach is a valuable QC option for high-throughput DNA/RNA isolations and could be adapted to additional fluorometric assays beyond nucleic acids.

Background: Sacroiliac joint (SIJ) pain is a common musculoskeletal condition among postpartum women due to hormonal, biomechanical, and physiological changes during pregnancy and childbirth. Increased ligament laxity, altered posture, weight gain, and pelvic instability may contribute to sacroiliac joint dysfunction. This condition often leads to pelvic girdle pain, low back pain, and functional limitations that affect daily activities such as walking, sitting, standing, and childcare. Objective: To determine the prevalence of sacroiliac joint pain and assess the level of disability among postpartum females in Sialkot. Materials and Methods: A descriptive cross-sectional study was conducted among 300 postpartum women aged 20-40 years from public and private hospitals in Sialkot. Non-probability convenience sampling approach was employed. Sacroiliac joint pain was assessed using clinical provocation tests including FABER, compression, and distraction tests. Pain intensity was measured using the Numerical Pain Rating Scale (NPRS), and functional disability was evaluated using the Oswestry Disability Index (ODI). Data were analyzed using SPSS version 25. Results: Clinical provocation testing demonstrated positive SIJ pain provocation in 84.0% (n=252) of acute postpartum participants, with severe functional disability observed in 46.0% (n=138). Most women reported moderate pain (74.7%), while 20.7% experienced severe pain. According to the ODI, 46% of participants had severe disability, 37.7% moderate disability, 10.7% minimal disability, and 5.7% were classified as crippled. Conclusion: Sacroiliac joint pain is highly prevalent among postpartum females and is associated with considerable functional disability. Early screening and physiotherapy-based rehabilitation may help reduce pain and improve functional outcomes. Keywords: Sacroiliac joint pain, Postpartum women, Pelvic girdle pain, Functional disability, Oswestry Disability Index.

Background Vitamin D may influence bone healing and recovery after spine surgery, but its clinical effects remain uncertain. Objective To evaluate clinical, functional, and bone-related outcomes associated with vitamin D status and supplementation in patients undergoing spine surgery. Methods This systematic review and meta-analysis was registered in PROSPERO (CRD420261467482) and reported according to PRISMA 2020. PubMed/MEDLINE, Scopus, Web of Science, Embase, and the Cochrane Library were searched from inception through 1 June 2026. Random-effects meta-analyses used mean differences (MDs) for continuous outcomes and risk ratios (RRs) for dichotomous outcomes. Direct supplementation analyses were prioritized, while analyses combining supplementation, baseline vitamin D status, or co-interventions were considered exploratory. Risk of bias was assessed using design-specific Joanna Briggs Institute tools. Results Thirteen studies were included. Four supplementation studies involving 177 participants showed no significant reduction in postoperative pain (MD -0.82, 95% CI -2.50 to 0.86; I2 = 84.3%). Three studies involving 277 participants showed lower Oswestry Disability Index scores in an exploratory analysis (MD -6.40, 95% CI -10.41 to -2.39; I2 = 96.8%). Three supplementation studies involving 128 participants showed no significant improvement in fusion rate (RR 1.17, 95% CI 0.78 to 1.74; I2 = 31.5%). An exploratory four-study fusion analysis was also not statistically significant (RR 1.24, 95% CI 0.98 to 1.57; I2 = 45.5%). Conclusion Current evidence does not establish that vitamin D supplementation independently improves postoperative pain or fusion rates after spine surgery. Possible benefits for disability and other bone-related outcomes remain uncertain because of small study numbers, heterogeneity, co-interventions, and residual confounding. Larger randomized trials stratified by baseline vitamin D status are needed.

Small molecule modifiers whence bound, allosterically, will alter the binding of a macromolecule to one- or more-cognate substrates/partners via conformational and non-conformational changes. Although allostery is inferred directly from empirical data, the mathematical basis of these models, constraints deployed and choice of parameter(s) are not clear. Here, we present and characterize a discrete-to-continuous mathematical model for ensemble distributions of a ligand-interacting macromolecular species across milieux-dependent conformational states and examine its role in the genesis and progression of cooperative binding. The premise, of our model, is a set of occupancy matrices (sparse, binary, strictly delocalized) which can be partitioned by a probability-based hyperparameter into mutually exclusive proper subsets of occupancy matrices with identical multinomial probabilities. Since each subset is canonical with a constituent occupancy matrix, it is characterized by a unique multinomial probability. The inner product of combinatorial pairs of all mutually exclusive subsets of occupancy matrices, with an expression for the summed transitional probabilities (finite differences between unique multinomial probabilities), is the differentiable matrix of strictly positive real-valued numbers for the system of ensemble distributions. Whilst the harmonic mean is presented as a generic solution for a system of ensemble distributions, the row-wise definite integral for each column is the finite union of open intervals (contiguous, strictly monotone) which in tandem with a set of interval-specific and bounded transitional probabilities constitutes a piecewise smooth curve (path-connected-, closed- and compact-set). Our discrete-to-continuous model is phenomenological and able to recapitulate the basic tenets of cooperative binding whilst offering insights into the genesis and progression of the same.

Myofascial pain syndrome (MPS) is characterized by increased muscle stiffness, trigger points, and functional limitations, yet clinical diagnosis remains largely subjective. Shear wave elastography (SWE) provides quantitative assessment of muscle mechanical properties, but its value for identifying biomechanical and clinical phenotypes of MPS is not fully established. This study evaluated whether stiffness parameters derived from multi-angle SWE can reliably characterize upper-trapezius anisotropy, and whether integrating SWE with bioimpedance spectroscopy (BIS), range of motion (ROM), and patient-reported outcomes (PROs) improves differentiation of MPS subgroups. Seventy-one adults completed upper-trapezius SWE, BIS, ROM assessments, and PRO measures. Clinically, 18 were classified as active MPS, 36 as latent, and 17 as normal. Shear wave speed measurements were modeled to estimate longitudinal (uL), transverse (uT), and anisotropy (uE) components. Reliability was examined using intraclass correlation coefficients. Unsupervised clustering and partial-correlation network analysis were applied to biomechanical and clinical variables. uT showed the strongest associations with BIS frequency parameters and ROM measures, indicating sensitivity to fascial composition, and mobility. Multimodal clustering incorporating uT with Fc or ROM identified subgroups with distinct tissue-level and functional characteristics. Network analysis demonstrated a progression in connectivity patterns, shifting from localized mechanical relationships to broader symptom-level coupling involving pain interference, sleep disturbance, emotional distress, and physical function. These findings indicate that SWE-derived stiffness parameters provide reliable, direction-specific quantification of trapezius mechanical properties. Combining SWE with impedance and mobility measures yields physiologically coherent MPS phenotypes that differ in both biomechanical features and clinical network structure, supporting more objective framework for characterizing MPS.

This protocol describes a standardized and reproducible minimally invasive approach for establishing mouse models of bisphosphonate-related osteonecrosis of the jaw (BRONJ) and osteoradionecrosis of the jaw (ORNJ). The method combines a unified low-trauma oral surgical procedure with disease-specific injury induction strategies to generate robust and clinically relevant models of jaw osteonecrosis. For BRONJ, systemic zoledronic acid administration is coupled with mandibular first molar extraction using tape-assisted mouth opening and customized bent micro-forceps, minimizing soft tissue damage and reducing procedural variability. For ORNJ, a customized lead-shielding platform enables precise, noninvasive mandible-targeted irradiation, producing reproducible bone injury while limiting off-target radiation exposure. Together, these complementary models provide a consistent and minimally invasive framework for investigating jaw osteonecrosis arising from distinct etiologies. The protocol supports comprehensive downstream analyses, including micro-computed tomography, histology, and immunofluorescence, and facilitates mechanistic studies of disease pathogenesis, bone regeneration, and therapeutic intervention.

Precise quantification of myofilament protein phosphorylation is essential for understanding the regulation of cardiac contractility in health and disease. Although Phos-tag SDS-PAGE is widely used to resolve phosphorylated protein isoforms, its reproducibility and quantitative reliability are often limited by variability in the key experimental factors, including gel composition, electrophoretic conditions, protein loading, and sample preparation. Here, we present a standardized manganese (Mn2+)-Phos-tag SDS-PAGE workflow optimized for cardiac myofilament proteins, using myosin regulatory light chain 2 (MLC2) and cardiac troponin I (cTnI) as model targets. We systematically evaluated critical parameters - including Mn2+ and Phos-tag concentrations, acrylamide composition, electrophoretic regime, buffer chemistry, protein loading, and EDTA-mediated transfer - to define conditions that maximize phospho-species resolution while preserving quantitative fidelity. We further demonstrate that electrophoresis rate, sample loading, and extraction strategy significantly influence band morphology, signal intensity, and the apparent distribution of phospho-species. As a use case scenario, we compared Trichloroacetic acid (TCA) extracted mouse left ventricular homogenates with myofibrils prepared using a widely adopted Triton-X-100 tissue-demembranization protocol. Myofibril preparation was associated with profound MLC2 dephosphorylation at the earliest stages of preparation, whereas cTnI exhibited a marked reduction in higher-order, low-stoichiometry phosphoforms. Further evaluation of Myosin-binding protein C (MyBP-C) showed progressive loss of phosphorylation over the course of 24 hours. We submit that TCA-extracted heart standards in combination with Phos-tag gels can provide valuable quality control for the phosphorylation status of myofibril preparations, and that inclusion of a high-affinity PP2A and PP1 phosphatase inhibitor like okadaic acid may benefit future myofibril mechanics studies.

Photoplethysmography (PPG, optical measurement of cardiac blood volume changes) is the foundation of wearable cardiac monitoring, but systematically fails on dark skin due to melanin absorption. We present the Melanin Absorption Invariance (MAI) framework: a label-free method that substantially reduces cross-skin-tone bias in cardiac feature extraction by preserving topological rather than geometric signal structure. We prove two theorems: Theorem 1 bounds attractor bias to O(SNR_eff^-1) under Z-normalization; Theorem 2 reduces residual bias to O(SNR_eff^-2) via SNR-adaptive correction. Empirical validation confirms these theoretical predictions on real dark-skin PPG signals. Comprehensive empirical validation on the complete MMPD dataset (Fitzpatrick III-VI, n = 656 recordings, 33 subjects, spanning all 4 lighting conditions and 5 motion types, Samsung Galaxy mobile phone) demonstrates MAI generalization across real-world deployment conditions. Results show substantial attractor bias reduction across all skin tone groups, with largest effects for Fitzpatrick IV and VI populations most affected by current systems. This work demonstrates a theoretically grounded, label-free, skin-tone-invariant cardiac monitoring framework.

Distal radius fractures (DRFs) can delay return to activities of daily living and social participation because of postoperative pain, temporary joint immobilization, and limited wrist and forearm range of motion. The Ghost System developed at Saitama Prefectural University, Japan, combines visual action observation with tendon vibration stimulation and has shown potential as an adjunct to conventional rehabilitation. This Study Protocol describes a modified Ghost system intended to improve clinical implementation by replacing the head-mounted virtual reality display with iPad-based action observation and by using a wristband-type vibrator. This single-center, single-arm, open-label feasibility trial will enroll 10 adults after palmar locking plate fixation for DRF. The intervention will be delivered twice weekly during outpatient rehabilitation follow-up sessions from the early postoperative period (postoperative days 2-10 after enrollment) through the approved early postoperative rehabilitation period (generally up to postoperative week 8), in parallel with standard rehabilitation practices. Primary feasibility and preliminary clinical outcomes include device fit and acceptability, pain assessed using a 100-mm Visual Analog Scale, and wrist/forearm range of motion. Secondary implementation and safety outcomes include Disabilities of the Arm, Shoulder and Hand (DASH), Patient-Rated Wrist Evaluation (PRWE), Hand20 Questionnaire (HANDS-20), EuroQol 5 Dimensions 5 Levels (EQ-5D-5L), body ownership and hand-illusion questionnaires, setup time, setup errors, adherence, adverse events, and device incidents. We hypothesize that the modified Ghost system will be feasible and acceptable for early postoperative outpatient rehabilitation and will be delivered without serious device-related adverse events. Clinical outcomes will be summarized descriptively to inform a future controlled study rather than to establish efficacy.

Background Bone-avid tracer myocardial scintigraphy for the diagnosis of transthyretin amyloid cardiomyopathy (ATTR-CM) has traditionally employed imaging at one or 3-hour intervals. Technetium-99m hydroxydiphosphonate (99mTc-HDP) has unique characteristics that may enable earlier imaging. We investigated the diagnostic concordance of immediate versus 1-hour acquisitions. Methods Consecutive patients with suspected ATTR-CM underwent planar imaging and SPECT/CT immediately and at 1-hour following the administration of 99mTc-HDP. Perugini grades and heart to contralateral lung (H/CL) ratios were assessed. Target-to-background ratios (TBRs) were calculated on the SPECT/CT acquisitions using the left ventricular (LV) septum and three background regions: aorta, LV blood-pool, and vertebrae. We assessed diagnostic concordance using Cohen's Kappa ({kappa}), temporal stability using paired t-tests, and correlation between timepoints using Pearson's coefficient (r). The 1-hour SPECT/CT interpretation served as the protocol reference standard. Results Forty-eight patients (83% male; median age, 80 [73-85] years) were evaluated. One-hour SPECT/CT identified 19 positive and 29 negative cases. Immediate SPECT/CT demonstrated 100% diagnostic concordance with the 1-hour reference standard ({kappa} = 1.000; 95% CI: 1.00 to 1.00; p < 0.001). The LV septum/LV Blood-Pool TBR showed the highest correlation (r = 0.956; 95% CI: 0.922 to 0.975; p < 0.001). The LV Septum/Aorta TBR demonstrated high correlation (r = 0.918; 95% CI: 0.857 to 0.953; p < 0.001) and remained stable in the ATTR-negative cohort (-0.02; 95% CI: -0.08 to 0.04; p = 0.54). Significant decrease in the LV Septum/Vertebrae TBR in the ATTR-negative (-0.55; 95% CI: -0.64 to -0.47; p < 0.001) and ATTR-positive cohorts (-1.14; 95% CI: -1.39 to -0.89; p < 0.001) was observed. Conclusions Immediate 99mTc-HDP SPECT/CT is diagnostically concordant with standard 1-hour protocols. By leveraging SPECT/CT and the favorable kinetics of 99mTc-HDP, immediate-phase imaging can accurately reproduce 1-hour acquisitions in cases of suspected ATTR-CM. This expedited approach may improve nuclear laboratory throughput and patient satisfaction.

Mechanical forces have a major effect on cell behavior. Most cells in vitro are grown under static conditions on hard tissue culture plastic, conditions that do not accurately reflect living tissues. The ability of cells to sense and respond to mechanical forces is essential for key biological processes, including development, proliferation, and migration. Disruption of the ability to respond to mechanical forces are known to be a critical factor in many diseases, including cardiovascular disease, progeria, and cancer. Here, we present the design, fabrication, and biological testing of a custom-built cell-stretching device that applies controlled biaxial strain to cells cultured on a polydimethylsiloxane (PDMS) membrane. We then used this device to examine how cells respond to strain. In response to biaxial strain, MCF-7 cells activated the mechanosensitive immediate early gene (IEX-1), with its expression increasing significantly after 1 and 3 hours of stretching. Cells exposed to mechanical strain also remodeled their cytoskeleton in a direction-dependent manner. Under uniaxial strain, actin filaments reoriented perpendicular to the stretch direction, whereas biaxially stretched cells do not promote directional reorientation, but instead appear to reinforce actin at the cell periphery. Similarly, cells under uniaxial strain exhibited changes in nuclear orientation and shape that were not observed under biaxial strain. Nuclear area remained unchanged in either strain condition. These results highlight that the biaxial stretcher can be used to apply strain to cells, and that cells respond differently to biaxial strain compared to what has been reported for uniaxial strain.

Background: Spinal cord injury (SCI) is frequently associated with orthostatic hypotension, defined by a sustained decrease in blood pressure upon assuming an upright posture due to impaired autonomic regulation. Cardiovascular spinal cord epidural stimulation (CV-scES) can regulate systolic blood pressure (SBP) in people with SCI, but stimulation paradigms are highly individualized. To make this treatment available to more patients, we developed an algorithm to tailor individualized CV-scES paradigms that closely mimic researcher-developed paradigms. Methods: We performed an offline analysis using datasets collected from eight individuals with SCI with epidural stimulators implanted over the lumbosacral spinal segments. During data collection, researchers modulated stimulation parameters with the goal of maintaining SBP between 110-120 mmHg. Each two-hour dataset included synchronized SBP and stimulation recordings. We ran optimization analyses offline to determine temporal requirements before modifying stimulation amplitude to mitigate out-of-range SBP. Results: The algorithm parameters that best matched researcher-selected stimulation changed relatively quickly during the first 12 min (one every ~40 sec), and more slowly thereafter (one every ~79 sec). Overall, algorithmic stimulation closely tracked researcher-controlled stimulation, with a mean correlation coefficient of 0.94. To evaluate online performance, we tested the algorithm in real time with a single participant. We found that a faster approach was needed to respond to changes in SBP caused by rapid, unpredictable events, such as postural changes. We implemented a sigmoid-based paradigm that determined the time to wait before changing stimulation as a function of the current SBP, with worse SBP values requiring faster responses. The new paradigm outperformed the original algorithm and researcher-controlled stimulation across measures of SBP stability, though recovery from a postural tilt maneuver remained slower than with researcher control. Conclusions: Our results indicate that algorithmic stimulation may minimize assistance required from researchers and participants, making CV-scES more feasible for clinical translation.

Background: Extreme environmental events, such as extreme temperatures and air pollution, have become a global concern due to their detrimental effects on human health. Short-term peak exposure episodes, despite lasting only a few hours, are crucial for exposure-response modelling. The use of time-aggregated exposure data often overlooks the impact of peak exposures on human health. However, studies employing high-temporal resolution exposure data are rare due to the limited availability of high-temporal resolution health outcomes across various scenarios. Therefore, to address the limitations associated with exposure-response modelling using aggregated exposure data, we have developed a model referred to as the mixed-frequency distributed lag non-linear model (mf-DLNM). Methods: In this work, a simulation study was conducted to further validate the mf-DLNM for hourly-daily mixed-frequency data, using data on hourly temperature and daily respiratory mortality for the West Midlands, UK. Given that the focus was on extreme exposures, Relative Risks (RR) at the 5th and 95th temperature quantiles were considered as the estimands of interest. Model performance was evaluated based on the bias, empirical standard error (EmpSE), and coverage of these estimands. Additionally, the model was assessed across various scenarios, considering data size (1, 3, 5, and 11 years with a 24-hour lag), lag length (12 and 24 hours with 11 years), seasonal variation (summer months with 11 years and 24-hour lag) and distribution (Poisson and negative binomial). Results: The mf-DLNM effectively captured the true parameters of the model. The model, fitted to 11 years of simulated data, a 24-hour lag and a Poisson distribution, observed a bias of 0.011 (0.0009) and 0.011 (0.001) for the RR at the 5th and 95th temperature quantiles, respectively, with Monte Carlo SEs (MCSEs) in parentheses. Furthermore, the model exhibited coverage of 0.94 and 0.93 for RR at the 5th and 95th temperature quantiles, respectively. In addition, the mf-DLNM with hourly and daily data demonstrated satisfactory performance across all scenarios except for the RR at 95th temperature quantiles in the seasonal analysis. Conclusions: Researchers are encouraged to adopt mf-DLNM in instances where high-temporal resolution exposure data are available alongside low-resolution health data. It serves as an alternative to traditional approaches that aggregate high-frequency exposure data. By preserving the temporal information of environmental exposures, mf-DLNM enables a more precise assessment of exposure-response relationships, thereby improving the accuracy and reliability of health risk estimates. This approach offers a promising opportunity for informed decision-making and the development of effective interventions for vulnerable populations and healthcare facilities to address short-term environmental episodes.

Background. Cardiovascular disease is a leading cause of death among women with breast cancer, and the 2026 ACC/AHA dyslipidemia guideline endorses coronary artery calcium (CAC) scoring to guide statin therapy before cardiotoxic treatment. Breast cancer patients routinely undergo staging 18F-fluorodeoxyglucose PET/CT, whose low-dose CT visualizes the coronary arteries, thus enabling CAC quantification at no additional cost or radiation. Methods. In this single-center retrospective study, consecutive women with newly diagnosed breast cancer undergoing staging 18F-FDG PET/CT (2009?2021) had semi-automated Agatston CAC scoring performed on the low-dose CT and were stratified by CAC presence (CAC-P) versus absence (CAC-A). We assessed a composite of cardiac diagnostic testing (stress testing, coronary CT angiography, invasive angiography), clinical events, and reclassification of statin eligibility per ACC/AHA guideline thresholds in a prevention-eligible subgroup. Results. Among 276 women (mean age 55.5 years; median follow-up 7.1 years), CAC was present in 68 (25%) but was clinically reported in only 5.4%. CAC-P was associated with more cardiac testing (34% vs 12%; age-adjusted hazard ratio 2.75, 95% CI 1.43?5.28) and, though underpowered, with more atherosclerotic events (7.4% vs 1.4%), but not with the all-cause composite. In the prevention-eligible subgroup (n=39), CAC scoring would have changed statin eligibility in 64%, initiating therapy in 62% of CAC-P women and supporting de-prescribing in 67% of CAC-A women. Conclusions. CAC can be feasibly quantified from staging PET/CT in women with breast cancer and would frequently reclassify statin eligibility at no additional cost or radiation, yet is rarely reported.

Abstract Background: Non-specific chronic low back pain (NSCLBP) is a major cause of disability worldwide and is associated with low-grade systemic inflammation. This study investigated the effects of aerobic exercise on inflammatory biomarkers, pain intensity, and quality of life among individuals with NSCLBP. Methods: In this parallel-group randomized controlled trial, 41 participants with NSCLBP were allocated to either an aerobic exercise plus health education group (n=21) or a health education-only control group (n=20). Participants in the intervention group completed supervised aerobic cycling three times weekly for 12 weeks. Outcome assessors and laboratory personnel were blinded to group allocation. Outcomes were measured at baseline, Week 8, and Week 12. Results: Interaction effects were observed for TNF- (p=0.046), IL-6 (p<0.001), hs-CRP (p<0.001), and pain intensity (p<0.001). Significant improvements were also observed across all WHOQOL-BREF quality-of-life domains (all p<0.05). After adjustment for baseline values and age, participants in the intervention group had significantly lower Week 12 IL-6 (p=0.013), hs-CRP (p<0.001), and pain intensity (p<0.001) than controls. No serious adverse events were reported. Conclusions: Aerobic exercise combined with health education produced greater improvements in inflammatory biomarkers, pain intensity, and quality of life than health education alone among individuals with NSCLBP. These findings support the integration of structured aerobic exercise into rehabilitation programmes for chronic low back pain. Keywords: Non-specific chronic low back pain; aerobic exercise; inflammation; IL-6; hs-CRP; pain intensity; quality of life; randomized controlled trial.

Manual analysis of skeletal muscle cross-sections is time-consuming and subject to error and user bias. To overcome these limitations, we developed and validated a semi-automated, quantitative, and reproducible image-analysis pipeline specifically tailored to quantify Pax7+ satellite cells, myonuclei, and cross-sectional area by fiber type. The workflow combines FIJI/ImageJ-based image preprocessing with CellProfiler, Cellpose, and a custom Python script to process and analyze immunohistological images of muscle tissue cross-sections. Outcomes include Pax7+ satellite cells and myonuclei quantified per fiber by fiber type, along with cross-sectional area, perimeter, and fiber type classification. This semi-automated approach provides a robust and efficient platform for high-throughput analysis of muscle tissue cross-sections from large datasets. O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=141 SRC="FIGDIR/small/729866v1_ufig1.gif" ALT="Figure 1"> View larger version (44K): org.highwire.dtl.DTLVardef@a3401dorg.highwire.dtl.DTLVardef@1c63145org.highwire.dtl.DTLVardef@ccbf76org.highwire.dtl.DTLVardef@2e0da0_HPS_FORMAT_FIGEXP M_FIG C_FIG

Spinal cord injury (SCI) causes rapid and severe bone loss in the paralysed lower limbs, particularly at the distal femur and proximal tibia, where fragility fracture risk is high. In vitro nanoscale vibration at 1 kHz has been shown to promote osteogenic differentiation and inhibit osteoclastogenesis, suggesting potential as a targeted mechanical intervention. This study aimed to develop and evaluate a wearable device for delivering and monitoring localised nanovibration at the distal femur in individuals with SCI. The device delivered continuous sinusoidal nanoscale stimulation at 1 kHz via a bone-conduction transducer, with an opposing accelerometer used to monitor transmitted vibration in real time. Design and target-site selection were refined through two healthy-volunteer investigations comparing the distal femur, proximal tibia, and distal tibia. Bovine femur experiments characterised vibration transmission under controlled benchtop conditions. Preliminary repeated-use feasibility was assessed in one individual with motor-complete SCI. Healthy volunteer testing showed that although the ankle initially produced the highest transmitted amplitudes, these were highly variable, and positioning was inconsistent. Within the knee region, the distal femur provided the most practical and repeatable site for a wearable application. In bovine femur experiments, scanning laser vibrometry demonstrated measurable vibration on the condylar surface opposite the transducer, and depth-resolved measurements confirmed that nanoscale vibration remained detectable within bone. A gel interface layer reduced the transmitted amplitude. In the feasibility evaluation, 61 sessions were completed over 14 weeks, with logged accelerometry confirming repeated nanoscale vibration transmission. These findings establish feasibility and support further device optimisation and translational studies.