Corrigendum: Non-invasive methods to assess muscle function in dogs: a scoping review.

[This corrects the article DOI: 10.3389/fvets.2023.1116854.].

Effect of center of rotation of angulation-based levelling osteotomy on instantaneous center of rotation ex vivo.

Cranial cruciate ligament rupture is a common cause of femorotibial instability in dogs. Despite numerous techniques being described for achieving joint stabilization, no consensus exists on the optimal management strategy. This ex vivo study utilized the path of the instantaneous center of rotation (ICR) to compare normal, pathological and treated joints. Fluoroscopic recordings of seven limbs from a previous study of canine stifle joint stability following center of rotation of angulation-based levelling osteotomy (CBLO) with and without hamstring loading were analyzed using least-squares approximation of the ICR and estimation of percentage gliding (vs. rolling) to determine if alterations in ICR path and gliding caused by CCL transection and following meniscal release could be normalized by CBLO. In intact joints, the ICR path was located mid-condyle, but this shifted significantly proximally and caudally following CCL transection and medial meniscal release (p < 0.007, p < 0.04). Hamstring loading resulted in qualitative and some quantitative improvements in joint movement based on percentage gliding movement analysis. The ICR path after CBLO remained significantly different to the intact location with or without a hamstring load (p < 0.02, p < 0.04), potentially consistent with CBLO aims of mild residual instability. CBLO resulted in percentage gliding characteristics not significantly different to intact joints (p > 0.08). Qualitative improvements in ICR path and percentage gliding quantities and variability suggest that hamstring loading positively influences joint biomechanics and that further investigation of this role ex vivo and clinically is warranted.

Muscle function assessment of the hindlimbs in healthy dogs using acoustic myography.

INTRODUCTION: Impaired muscle function is a frequent consequence of musculoskeletal disorders in dogs. Musculoskeletal disorders, especially stifle joint diseases, are common in dogs and assessment of muscle function in dogs is clinically relevant. Acoustic myography (AMG) is a non-invasive method to assess muscle activity. Quantifying muscle function in normal dogs could help identify clinically relevant changes in dogs with orthopaedic disease and allow targeted interventions to improve recovery in these. The objectives of the study were to characterize hindlimb muscle function in healthy dogs using AMG and to investigate the repeatability and reproducibility of AMG in dogs. METHODS: Healthy dogs (15-40 kg) without musculoskeletal disorders were recruited and screened for eligibility to participate in the study. The muscle activity in four hindlimb muscles related to the stifle was assessed using AMG. The degree of symmetry between the hindlimbs in these dogs was investigated and the reliability of AMG was evaluated. RESULTS AND CONCLUSIONS: The study population comprised 21 dogs. Reference intervals and symmetry indices for AMG scores of the hindlimb muscles were identified, with highest variability for the E-scores. For all AMG-scores, same-day variation was lower than between days variation, and both were lowest for S- and T-scores. Further investigation is needed to establish if AMG can enable discrimination between dogs with altered muscle function and healthy dogs.

Cranial closing wedge ostectomies for management of canine cranial cruciate ligament insufficiency: Comparison and geometric modelling of errors.

Variants of the cranial closing wedge ostectomy (CCWO) may be used to manage cranial cruciate ligament deficient stifles by reduction of the tibial plateau angle (TPA). Concerns have been raised about predictability of achieving target TPA, limb shortening, axis shift, and distalization of the tibial tuberosity. Landmarks derived from tibial radiographs (n = 50) by 5 observers, along with outlines of the cranial and caudal proximal tibial cortices, were used to simulate six CCWO variants. Tested variants were Slocum-type (n = 2), modified (n = 2), isosceles, and anatomical-mechanical axis (AMA)-based CCWO. Observer specific landmarks were used to model pre- and postoperative errors in TPA measurement. Mean landmarks were used to compare key outcome variables between the variants, as well as modelling errors in positioning the wedge apex. Variants differed markedly in TPA, axis shift, mechanical axis length changes, and tibial tuberosity distalization, with more proximally positioned wedges performing better than distally positioned wedges. Errors in TPA identification appeared partially self-correcting due to altered axis shifts. Craniocaudal shifts in wedge apex position had the most severe effects on outcome variables, especially in shorter tibiae with greater initial TPA values. Recent CCWO variants are likely to achieve a satisfactory postoperative TPA with limited axis shift. The AMA-based CCWO technique appears to influence MA length and tibial tuberosity position the least, and apart from the effect on TPA where no advantage could be discerned, it appeared more robust in the face of wedge apex positioning errors than the other techniques.

Diagnosis of medial meniscal lesions in the canine stifle using multidetector computed tomographic positive-contrast arthrography.

OBJECTIVE: To assess diagnostic value and clinical utility of multidetector computed tomographic positive contrast arthrography (CTA) for meniscal lesions in dogs. STUDY DESIGN: Prospective case series. STUDY POPULATION: Client-owned dogs (n = 55) with cranial cruciate ligament injuries. METHODS: Sedated dogs underwent CTA using a 16-slice scanner, and subsequently received mini-medial arthrotomy for meniscal assessment. Scans were anonymized, randomized, and reviewed twice for meniscal lesions by three independent observers with varying experience. Results were compared with surgical findings. Reproducibility and repeatability were assessed with kappa statistics, intraobserver changes in diagnosis by McNemar's test, and interobserver differences using Cochran's Q test. Test performance was calculated using sensitivity, specificity, proportion correctly identified, and positive and negative predictive values and likelihood ratios. RESULTS: Analysis was based on 52 scans from 44 dogs. Sensitivity for identifying meniscal lesions was 0.62-1.00 and specificity was 0.70-0.96. Intraobserver agreement was 0.50-0.78, and interobserver agreement was 0.47-0.83. There was a significant change between readings one and two for the least experienced observers (p < .05). The sum of sensitivity and specificity exceeded 1.5 for both readings and all observers. CONCLUSION: Diagnostic performance was suitable for identifying meniscal lesions. An effect of experience and learning was seen in this study.

Objective and owner-reported outcomes after modified cranial closing wedge ostectomy: a case series.

Immediate and longer-term outcomes of a cranial closing wedge ostectomy variant for management of canine cranial cruciate ligament disease were assessed in this single-center retrospective consecutive study. Records and radiographs were retrieved and assessed by three independent observers to evaluate tibial plateau angle, anatomical-mechanical axis angle, tibial tuberosity distalization, and mechanical axis length before and after surgery. Kinetic gait analysis and owner questionnaires were used to assess clinical outcomes. Seventeen stifles from fifteen dogs were evaluated radiographically. Mean error from target tibial plateau angle was 0.4 degrees. Anatomical-mechanical axis angles reduced from mean 2.9 degrees preoperatively to mean - 0.9 degrees postoperatively. Tibial tuberosity distalization was mean 5.0% of mechanical axis length, and mean reduction in mechanical axis length was 0.1%. Increased tibial plateau angles were noted in 8/17 stifles, with a mean of 9.6 degrees at short-term follow-up. Major complications were observed in 9/17 stifles. Long term follow-up (mean 832 days) was obtained with gait analysis in 8/15 dogs and with questionnaire in 11/15. Most dogs (9/11) were weakly to moderately affected by osteoarthritis symptoms. All values for peak vertical force and vertical impulse normalized to body weight exceeded local lower reference limits for normal dogs, indicating acceptable limb use. Satisfactory immediate and long-term clinical outcomes appear to be possible with this technique, but the high incidence of shorter-term complications may caution against the technique or the fixation and management described here.

Habitat type drives the spatial distribution of Australian fish chorus diversitya).

Fish vocalize in association with life functions with many species calling en masse to produce choruses. Monitoring the distribution and behavior of fish choruses provides high-resolution data on fish distribution, habitat use, spawning behavior, and in some circumstances, local abundance. The purpose of this study was to use long-term passive acoustic recordings to obtain a greater understanding of the patterns and drivers of Australian fish chorus diversity at a national scale. This study detected 133 fish choruses from year-long recordings taken at 29 Australian locations with the highest fish chorus diversity identified at a site in the country's northern, tropical waters. A linear model fitted with a generalized least squares regression identified geomorphic feature type, benthic substrate type, and northness (of slope) as explanatory variables of fish chorus diversity. Geomorphic feature type was identified as the significant driver of fish chorus diversity. These results align with broad-scale patterns reported previously in fish biodiversity, fish assemblages, and fish acoustic diversity. This study has highlighted that passive acoustic monitoring of fish chorus diversity has the potential to be used as an indicator of fish biodiversity and to highlight habitats of ecological importance.

The influence of flow velocity on the response of rheophilic fish to visual cues.

The strong association with visual cues exhibited by fish that prefer to inhabit flowing water (rheophilic species) may help reduce the energetic costs of maintaining position due to the provision of spatial points of reference. If this "Station Holding Hypothesis" is true, a positive relationship between the association with visual cues and flow velocity is expected. This hypothesis was tested experimentally by quantifying the response of common minnow (Phoxinus phoxinus) and brown trout (Salmo trutta) to visual cues under three flow velocities. In contradiction to the prediction, there was no evidence that the association with strong visual cues was positively related to flow velocity when fish were presented with vertical black stripes in an open channel flume, although interspecific variation in response was observed. The association with visual cues was relatively weak in trout, compared to minnow that spent 660% more time associated with the zone in which visual cues were present during the treatment, than the control when visual cues were absent. Trout tended to be more exploratory and made short visits to the area where visual cues were present, whereas minnow associated with the cues for longer. The strong association with visual cues independent of flow velocity exhibited by minnow and the weak association across all velocities by trout suggest that this behaviour is unlikely to reflect a strategy to minimise the energetic cost of maintaining position in flowing water. Minnow may have used the visual cues as a proxy indicator of physical structure that provides alternative benefits, such as refuge from predators. Trout may have employed alternative cues (e.g. mechanosensory) to seek more energetically favourable regions of the experimental area, reducing the importance of stationary visual stimuli.

Non-invasive methods to assess muscle function in dogs: A scoping review.

Skeletal muscle function can be affected by multiple disorders in dogs of which cranial cruciate ligament rupture or disease (CCLD) is one of the most common. Despite the significance of this condition only sparse research exists regarding assessment of muscle function in dogs. This scoping review aimed to identify the non-invasive methods for canine muscle function assessments that have been reported in the literature in the past 10 years. A systematic literature search was conducted 1st March 2022 across six databases. After screening, 139 studies were considered eligible for inclusion. Among the included studies, 18 different muscle function assessment categories were identified, and the most frequently reported disease state was CCLD. We included an attempt to elucidate the clinical applicability of the 18 reported methods, as experts were asked to subjectively assess the methods for their clinical relevance as well as their practical applicability in dogs with CCLD.

Functional Engagement of the PD-1/PD-L1 Complex But Not PD-L1 Expression Is Highly Predictive of Patient Response to Immunotherapy in Non-Small-Cell Lung Cancer.

PURPOSE: In many cancers, the expression of immunomodulatory ligands leads to immunoevasion, as exemplified by the interaction of PD-L1 with PD-1 on tumor-infiltrating lymphocytes. Profound advances in cancer treatments have come with the advent of immunotherapies directed at blocking these immuno-suppressive ligand-receptor interactions. However, although there has been success in the use of these immune checkpoint interventions, correct patient stratification for these therapies has been challenging. MATERIALS AND METHODS: To address this issue of patient stratification, we have quantified the intercellular PD-1/PD-L1 interaction in formalin-fixed paraffin-embedded tumor samples from patients with non-small cell lung carcinoma, using a high-throughput automated quantitative imaging platform (quantitative functional proteomics [QF-Pro]). RESULTS: The multisite blinded analysis across a cohort of 188 immune checkpoint inhibitor-treated patients demonstrated the intra- and intertumoral heterogeneity of PD-1/PD-L1 immune checkpoint engagement and notably showed no correlation between the extent of PD-1/PD-L1 interaction and PD-L1 expression. Importantly, PD-L1 expression scores used clinically to stratify patients correlated poorly with overall survival; by contrast, patients showing a high PD-1/PD-L1 interaction had significantly better responses to anti-PD-1/PD-L1 treatments, as evidenced by increased overall survival. This relationship was particularly strong in the setting of first-line treatments. CONCLUSION: The functional readout of PD-1/PD-L1 interaction as a predictive biomarker for the stratification of patients with non-small-cell lung carcinoma, combined with PD-L1 expression, should significantly improve the response rates to immunotherapy. This would both capture patients excluded from checkpoint immunotherapy (high PD-1/PD-L1 interaction but low PD-L1 expression, 24% of patients) and additionally avoid treating patients who despite their high PD-L1 expression do not respond and suffer from side effects.

Ideal Anchor Points for Patellar Anti-rotational Sutures for Management of Medial Patellar Luxation in Dogs: A Radiographic Survey.

OBJECTIVE: The aim of this study was to identify the ideal anchor point for patellar anti-rotational sutures for adjunctive stabilization of medial patellar luxation in both small and large breed dogs. STUDY DESIGN: Retrospective radiographic survey was performed on 110 stifles from 101 dogs. Radiographs were grouped based on patient weight (≤15 kg; >15 kg) and diagnosis (medial patellar luxation, cranial cruciate ligament rupture, and normal joints). Radiographic measurements included: the proximal, middle, and distal points of the trochlear ridge, the caudal aspect of Blumensaat's line (roof of the intercondylar notch), the centre of the lateral fabella, as well as the "best-fit" centre of a circle overlying the trochlea. These landmark coordinates were used to calculate radii for comparison, and for scaling between joints. RESULTS: Use of the fabellar centre resulted in larger radii (corrected p <0.001) than those from the best-fit circle centre for all but one combination of patient group and trochlear end point locations. Using the best-fit circle centre, radius variation was less marked than with the fabellar centre. Significant differences in location for centres of the best-fit circle and fabella were seen across all patient categories (Pillai's trace p <0.001). CONCLUSION: The fabella is unlikely to be the best choice for anchoring a patellar anti-rotational suture. Use of the best-fit circle centre to place a suture anchor should be preferred to maximise suture isometry during joint flexion and extension in large and small breed dogs.

Determination of Interactive States of Immune Checkpoint Regulators in Lung Metastases after Radiofrequency Ablation.

BACKGROUND: Cases of the spontaneous regression of multiple pulmonary metastases, after radiofrequency ablation (RFA), of a single lung metastasis, have been documented to be mediated by the immune system. The interaction of immune checkpoints, e.g., PD-1/PD-L1 and CTLA-4/CD80, may explain this phenomenon. The purpose of this study is to identify and quantify immune mechanisms triggered by RFA of pulmonary metastases originating from colorectal cancer. METHODS: We used two-site time-resolved Förster resonance energy transfer as determined by frequency-domain FLIM (iFRET) for the quantification of receptor-ligand interactions. iFRET provides a method by which immune checkpoint interaction states can be quantified in a spatiotemporal manner. The same patient sections were used for assessment of ligand-receptor interaction and intratumoral T-cell labeling. CONCLUSION: The checkpoint interaction states quantified by iFRET did not correlate with ligand expression. We show that immune checkpoint ligand expression as a predictive biomarker may be unsuitable as it does not confirm checkpoint interactions. In pre-RFA-treated metastases, there was a significant and negative correlation between PD-1/PD-L1 interaction state and intratumoral CD3+ and CD8+ density. The negative correlation of CD8+ and interactive states of PD-1/PD-L1 can be used to assess the state of immune suppression in RFA-treated patients.

The fusion of quantitative molecular proteomics and immune-oncology: a step towards precision medicine in cancer therapeutics.

Innate and adaptive immune systems are built-in homeostatic functions of many multicellular organisms and protect the host against foreign pathogens and infections. Dysregulation of the molecular mechanisms of the immune system can result in autoimmune diseases. The immune system can also be harnessed and manipulated to provide targeted cancer therapies, some of them relying on the blockade of immune-checkpoint receptors. Two prominent immune checkpoints, PD-1/PD-L1 and CTLA-4/CD80, comprise receptor-ligand pairs that prevent the host immune cells from attacking host tissues. However, cancer cells upregulate the respective PD-L1 and CD80 ligands for PD-1 and CTLA-4 and thereby evade the host-immune response. Therapeutic drugs that block PD-1/PD-L1 and CTLA-4/CD80 interactions re-enable the immune system to attack cancer cells, but their prognostic biomarker remains challenging. In this review, we discuss how the use of quantitative molecular imaging can be exploited to predict the response to anti-PD-1/PD-L1 therapies and to identify cancer patients who would benefit from them.

Effect of center of rotation of angulation-based leveling osteotomy on ex vivo stifle joint stability following cranial cruciate ligament transection and medial meniscal release with and without a hamstring load.

OBJECTIVE: To evaluate the effect of center of rotation of angulation (CORA)-based leveling osteotomy (CBLO) and hamstring load on stifle stability following cranial cruciate ligament transection (CCLx) and medial meniscal release (MMR). STUDY DESIGN: Ex vivo experimental study. SAMPLE POPULATION: Cadaver hind limb preparations (n = 7). METHODS: After instrumentation, constant quadriceps and gastrocnemius loads with an optional hamstring load in a 3:1:0.6 ratio were applied, and stifles were extended from fully flexed using an electrical motor during fluoroscopic recording. The recording process was repeated after each of CCLx, MMR and CBLO and the extracted landmark coordinates were used for calculation of cranial tibial translation (CTT) and patellar ligament angle (PTA). RESULTS: Mean initial tibial plateau angle was 28.1°: post-CBLO the mean was 9.7°. Cranial tibial translation developed from 50° and 75° with CCLx and MMR respectively (p < .04, < .02) without hamstring loading. Hamstring loading mitigated CTT due to CCLx and delayed CTT until 120° for MMR (P < .02) in this model. CBLO prevented CTT, except at 140° without hamstring loading (P = .01). Similar results were seen for PTA, but CBLO curves were parallel to and lower than intact values at all tested angles (P < .04), consistent with induced effective joint flexion. CONCLUSION: CBLO to a target tibial plateau angle of 10° largely eliminated CTT induced by CCLx and MMR. Hamstring loads of 20% quadriceps load improved stifle stability in this model. IMPACT: Stifle stability following CBLO appears to be multifactorial and depends on meniscal integrity, joint angle, and hamstring strength.

Quantification of protein-protein interactions and activation dynamics: A new path to predictive biomarkers.

Oncogene dysregulation is a driver of neoplasia development and progression. The use of quantitative molecular imaging to quantify oncogene activation will be crucial in developing companion diagnostics which can identify personalised patient regimens. However, the evaluation of oncogene activation does not necessarily correlate with oncoprotein activation. Post-translational modifications, such as phosphorylation, lipidation and methylation, may enhance oncoprotein functionality. It is this functionality that progresses neoplasia and may correlate with patient outcome. Advanced molecular imaging may be used to directly quantify oncoprotein, as opposed to oncogene, activation. Time-resolved Förster Resonance Energy Transfer (TR-FRET) involves the non-radiative transfer of energy from one chromophore to another over distances of 1-10 nm; allowing FRET to be used as a "chemical ruler". TR-FRET can be utilised to directly elucidate spatial oncoprotein activation in single cells and patient tissues. In single cells, TR-FRET has uncovered the mechanisms by which PKCß1 is trafficked to the nucleus and cleaved. Additionally it has revealed the mechanism of activation of Akt/PKB, whereby Akt/PKB undergoes a conformational change, allowing the Thr308 site to be phosphorylated by PDK1. Moreover TR-FRET has been utilised to quantify HER2-HER heterodimerisation and Akt/PKB activation states in patient biopsies, where it is shown to be predictive of outcome/relapse. The role of TR-FRET is not solely limited to intracellular signalling events. A study has used TR-FRET to measure intercellular immune-checkpoint receptor-ligand interactions. Within this study it was seen that PD-L1 expression was not indicative of PD-1/PD-L1 interaction states in a range of solid tumours. Crucially, in melanoma and NSCLC, PD-1/PD-L1 interaction was a predictive of an improved patient outcome. PD-L1 expression did not predict patient outcome. Several groups have worked to improve Fluorescence lifetime imaging microscopy (FLIM) acquisition times, including the use of: window-galvanometers; multifocal multiphoton FLIM and parallel pixel excitation coupled with wide-field time-gated FLIM. The development of novel quantitative molecular imaging will be critical in the development of personalised patient therapies in the future.

Immunogenomics of Colorectal Cancer Response to Checkpoint Blockade: Analysis of the KEYNOTE 177 Trial and Validation Cohorts.

BACKGROUND & AIMS: Colorectal cancer (CRC) shows variable response to immune checkpoint blockade, which can only partially be explained by high tumor mutational burden (TMB). We conducted an integrated study of the cancer tissue and associated tumor microenvironment (TME) from patients treated with pembrolizumab (KEYNOTE 177 clinical trial) or nivolumab to dissect the cellular and molecular determinants of response to anti- programmed cell death 1 (PD1) immunotherapy. METHODS: We selected multiple regions per tumor showing variable T-cell infiltration for a total of 738 regions from 29 patients, divided into discovery and validation cohorts. We performed multiregional whole-exome and RNA sequencing of the tumor cells and integrated these with T-cell receptor sequencing, high-dimensional imaging mass cytometry, detection of programmed death-ligand 1 (PDL1) interaction in situ, multiplexed immunofluorescence, and computational spatial analysis of the TME. RESULTS: In hypermutated CRCs, response to anti-PD1 immunotherapy was not associated with TMB but with high clonality of immunogenic mutations, clonally expanded T cells, low activation of Wnt signaling, deregulation of the interferon gamma pathway, and active immune escape mechanisms. Responsive hypermutated CRCs were also rich in cytotoxic and proliferating PD1+CD8 T cells interacting with PDL1+ antigen-presenting macrophages. CONCLUSIONS: Our study clarified the limits of TMB as a predictor of response of CRC to anti-PD1 immunotherapy. It identified a population of antigen-presenting macrophages interacting with CD8 T cells that consistently segregate with response. We therefore concluded that anti-PD1 agents release the PD1-PDL1 interaction between CD8 T cells and macrophages to promote cytotoxic antitumor activity.

Quantification of biomarker functionality predicts patient outcomes.

Implementation of a quantitative molecular imaging method (iFRET), which determines receptor-ligand interactions, has led to the finding that patients with a low extent of PD-1/PD-L1 interaction in metastatic NSCLC, and malignant melanoma, display significantly worsened overall survival compared to those with a high level of interaction.

Quantifying the variability between multiple multiplanar reconstructions of computed tomography scans.

BACKGROUND: Multiplanar reconstructions of computed tomography (CT) scans can alleviate issues with bone or joint positioning during scan acquisition. The repeatability of these reconstructions is dependent on human operators applying reconstruction criteria, and therefore is subject to error, which could affect measurement reliability for angular or spatial measurements made for orthopaedic surgery. We describe a method for quantifying inter-reconstruction variability numerically and graphically using metadata from the CT header to find vectors describing reconstruction axis alignment. The approach is demonstrated using 3 sets of computed tomographic reconstructions of 24 vulpine femorotibial joints. RESULTS: Vectors describing axis alignments permitted identification and subsequent analysis of deviations from optimal alignment between reconstruction sets. For the worked example, alignment deviations equivalent to femoral abduction/adduction were nearly twice those for extension/flexion, and simulation of the effects of these deviations on measurements closely matched published data. CONCLUSIONS: The method presented here is straightforward and permits numerical and graphical analysis of reconstruction variability. Reconstruction alignment variability should be considered before adopting new reconstruction criteria for clinical use, and evaluated whenever there is suspicion that reconstruction variability could unduly influence subsequent measurements. These evaluations may help drive improvements in reconstruction criteria. The methods described here could also be employed for comparing patient positioning between scans and between different scan modalities.

Kinetic gait analysis in healthy dogs and dogs with osteoarthritis: An evaluation of precision and overlap performance of a pressure-sensitive walkway and the use of symmetry indices.

In veterinary practice, a thorough gait examination is essential in the clinical workup of any orthopedic patient, including the large population of dogs with chronic pain as a result of osteoarthritis. The traditional visual gait examination is, however, a subjective discipline, and systems for kinetic gait analysis may potentially offer an objective alternative for gait assessment by the measurement of ground reaction forces. In order to avoid unnecessary testing of patients, a thorough, stepwise evaluation of the diagnostic performance of each system is recommended before clinical use for diagnostic purposes. The aim of the study was to evaluate the Tekscan pressure-sensitive walkway system by assessing precision (agreement between repetitive measurements in individual dogs) and overlap performance (the ability to distinguish dogs with lameness due to osteoarthritis from clinically healthy dogs). Direction of travel over the walkway was investigated as a possible bias. Symmetry indices are commonly used to assess lameness by comparing ground reaction forces across different combinations of limbs in each dog. However, SIs can be calculated in several different ways and specific recommendations for optimal use of individual indices are currently lacking. Therefore the present study also compared indices in order to recommend a specific index preferable for future studies of canine osteoarthritis. Forty-one clinically healthy dogs and 21 dogs with osteoarthritis were included in the study. High precision was demonstrated. The direction of travel over the walkway was excluded as a possible bias. A significant overlap was observed when comparing ground reaction forces measured in dogs with osteoarthritis compared to clinically healthy dogs. In some affected dogs, symmetry indices comparing contralateral limbs differed from clinically healthy dogs, but in general, the overlap performance was insufficient and, consequently, general use of this method for diagnostic purposes in dogs with osteoarthritis cannot be recommended.