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Client compliance with prescribed medication instructions to treat their pets is a concern. This study describes factors associated with the noncompliance of dog owners with veterinary recommendations for medication, as well as client-reported barriers and aids to administering medications. A cross-sectional survey of dog owners' compliance with veterinary medication recommendations was performed from 9 January 2019 to 18 July 2020. A convenience sample of owners who prescribed medication for their dogs during or following elective veterinary examination was surveyed regarding medication administration experience and compliance. Owners were followed up to determine if the course of medication had been completed. Compliance data were analyzed descriptively. Logistic regression was performed with compliance as the outcome. Medication noncompliance was recorded for 47% (71/151) of owners. Increasing dog age was associated with better owner compliance (p < 0.05). Pet owners who used "nothing" as an aid to medicating were less likely to be noncompliant (p < 0.05). Forty-seven percent (71/151) of owners reported that "nobody" showed them how to administer the medication. One-third of dog owners (47/151) reported challenges in medicating their pets. The most common reason cited by clients reporting challenges was a resistant pet. Demonstration of medication administration techniques and discussion about available aids to medicating a pet may improve client compliance.

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Understanding causes of insect population declines is essential for the development of successful conservation plans, but data limitations restrict assessment across spatial and temporal scales. Museum records represent a source of historical data that can be leveraged to investigate temporal trends in insect communities. Native lady beetle decline has been attributed to competition with established alien species and landscape change, but the relative importance of these drivers is difficult to measure with short-term field-based studies. We assessed distribution patterns for native lady beetles over 12 decades using museum records, and evaluated the relative importance of alien species and landscape change as factors contributing to changes in communities. We compiled occurrence records for 28 lady beetle species collected in Ohio, USA, from 1900 to 2018. Taxonomic beta-diversity was used to evaluate changes in lady beetle community composition over time. To evaluate the relative influence of temporal, spatial, landscape, and community factors on the captures of native species, we constructed negative binomial generalized additive models. We report evidence of declines in captures for several native species. Importantly, the timing, severity, and drivers of these documented declines were species-specific. Land cover change was associated with declines in captures, particularly for Coccinella novemnotata which declined prior to the arrival of alien species. Following the establishment and spread of alien lady beetles, processes of species loss/gain and turnover shifted communities toward the dominance of a few alien species beginning in the 1980s. Because factors associated with declines in captures were highly species-specific, this emphasizes that mechanisms driving population losses cannot be generalized even among closely related native species. These findings also indicate the importance of museum holdings and the analysis of species-level data when studying temporal trends in insect populations.

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The inhibitory-kappaB kinases (IKKs) IKKα and IKKß play central roles in regulating the non-canonical and canonical NF-κB signalling pathways. Whilst the proteins that transduce the signals of each pathway have been extensively characterised, the clear dissection of the functional roles of IKKα-mediated non-canonical NF-κB signalling versus IKKß-driven canonical signalling remains to be fully elucidated. Progress has relied upon complementary molecular and pharmacological tools; however, the lack of highly potent and selective IKKα inhibitors has limited advances. Herein, we report the development of an aminoindazole-pyrrolo[2,3-b]pyridine scaffold into a novel series of IKKα inhibitors. We demonstrate high potency and selectivity against IKKα over IKKß in vitro and explain the structure-activity relationships using structure-based molecular modelling. We show selective target engagement with IKKα in the non-canonical NF-κB pathway for both U2OS osteosarcoma and PC-3M prostate cancer cells by employing isoform-related pharmacodynamic markers from both pathways. Two compounds (SU1261 [IKKα Ki = 10 nM; IKKß Ki = 680 nM] and SU1349 [IKKα Ki = 16 nM; IKKß Ki = 3352 nM]) represent the first selective and potent pharmacological tools that can be used to interrogate the different signalling functions of IKKα and IKKß in cells. Our understanding of the regulatory role of IKKα in various inflammatory-based conditions will be advanced using these pharmacological agents.

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INTRODUCTION: Alopecia areata (AA) is a disease that manifests as patchy hair loss on the scalp and other parts of the body; severe disease may result in disfigurement, functional impairment, and significant psychological distress. This condition is understood to be caused by autoimmunity to the hair follicle and subsequent arrest of hair growth. New medications, baricitinib and ritlecitinib, belong to the Janus kinase (JAK) inhibitor family and are among the first FDA-approved treatments for severe AA. In this manuscript, we aim to answer the question: What treatment options exist for AA in the military health care system (MHS)? In doing so, we review the pathogenesis, physical and psychosocial impact of AA, conventional treatment of AA, and the efficacy and safety of baricitinib and ritlecitinib. METHODS: A literature search was performed using PubMed, Embase, and Ovid for the history and pathogenesis of AA, psychosocial impact of disease, functional impairments, and current treatments. Keywords "alopecia areata," "current therapy for alopecia areata," "pathogenesis alopecia areata," "baricitinib," "ritlecitinib," "JAK inhibitor alopecia," "JAK inhibitor safety," "baricitinib efficacy," "alopecia eyelash," "alopecia nails," and "psychosocial impact of alopecia" were used for the search. The TRICARE manual was searched for guidelines applicable to the treatment of AA, DoD Instruction 6130.03 Volume 2 for medical standards for military service, and the U.S. Central Command Modification 15 for fitness of deployment to Central Command area of operations. RESULTS: Traditional treatments such as intralesional steroids may be effective for some patients, but difficulty lies in controlling extensive or refractory disease. Janus kinase inhibitors, baricitinib and ritlecitinib, are found effective at improving severe refractory disease; baricitinib induced hair regrowth in 32.6% more patients than placebo, and ritlecitinib was found to be superior to placebo by at least 24%. Currently, there is no coverage for therapeutic treatment of hair growth in the MHS. Additionally, military members are disqualified for continued service if they require immunomodulator medications such as baricitinib and ritlecitinib. Those on immunomodulators are unable to deploy worldwide. CONCLUSIONS: Baricitinib and ritlecitinib are effective treatments for widespread, progressive, and refractory AA. Although JAK inhibitors demonstrate improved effectiveness compared to non-immunomodulator treatments, their use in the MHS for this purpose is limited.

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Osteoarthritis is a leading cause of lameness and joint disease in horses. A simple, economical, and accurate diagnostic test is required for routine screening for OA. This study aimed to evaluate infrared (IR)-based synovial fluid biomarker profiling to detect early changes associated with a traumatically induced model of equine carpal osteoarthritis (OA). Unilateral carpal OA was induced arthroscopically in 9 of 17 healthy thoroughbred fillies; the remainder served as Sham-operated controls. The median age of both groups was 2 years. Synovial fluid (SF) was obtained before surgical induction of OA (Day 0) and weekly until Day 63. IR absorbance spectra were acquired from dried SF films. Following spectral pre-processing, predictive models using random forests were used to differentiate OA, Sham, and Control samples. The accuracy for distinguishing between OA and any other joint group was 80%. The classification accuracy by sampling day was 87%. For paired classification tasks, the accuracies by joint were 75% for OA vs. OA Control and 70% for OA vs. Sham. The accuracy for separating horses by group (OA vs. Sham) was 68%. In conclusion, SF IR spectroscopy accurately discriminates traumatically induced OA joints from controls.

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BACKGROUND: Studies on equine air transport practices and consequences are scarce. This prospective study aimed to describe horse and air journey details and practices, document how horse behavior and health changed during the air transport phases, quantify the occurrence of welfare issues, and identify possible associations between horse and journey details, air transport practices, and welfare issues. RESULTS: Data were collected from before departure to five days after arrival on 118/597 horses traveling on 32 commercial air journeys on different routes, varying in duration and conditions. Most horses were middle-aged warmblood females, 26% of which were pregnant, and being moved by air for sales. Before flying, most were quarantined (median: 18; IQR: 9-53 days), and their fitness for travel was certified by veterinarians. At the departure airports, external temperatures varied from - 6 °C to 33 °C, and horses were loaded by experienced flight grooms (median: 35; IQR: 15-40 years) into jet stalls (three-horse: 87%, two-horse: 13%). During the flights, horses were regularly watered (water intake median: 14 L) and fed ad libitum (feed consumption median: 8 kg). At the arrival airport, horses were unloaded from the jet stalls, and external temperatures ranged from - 5 °C to 32 °C. Then, all horses were transported to arrival quarantine by road. Air transport phases affected horses' health status and behavior; increased heart and respiratory rates and behaviors, such as pawing, head tossing, and vocalization, were mainly identified at departure and arrival. Horse interaction, nasal discharge, increased capillary refill time (CRT), and abnormal demeanor were observed more often one hour before landing while resting and normal capillary refill time were more often displayed five days after arrival (all P < 0.01). One hour before landing, horses with bad temperament and horses of unknown temperament were more likely to develop nasal discharge when transported in winter and autumn (P < 0.001). The likelihood of an increased CRT was associated with shorter flights in horses of unknown travel experience (P < 0.001). Ten horses were injured, and 11 developed pleuropneumonias (i.e., shipping fever). CONCLUSIONS: Air transport is a complex procedure with several different phases affecting horse health and behavior. Therefore, experienced staff should carefully manage each horse before, during, and after air journeys to minimize welfare hazards.

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Since its discovery, a major debate about mitochondrial uncoupling protein 3 (UCP3) has been whether its metabolic actions result primarily from mitochondrial inner membrane proton transport, a process that decreases respiratory efficiency and ATP synthesis. However, UCP3 expression and activity are induced by conditions that would seem at odds with inefficient 'uncoupled' respiration, including fasting and exercise. Here, we demonstrate that the bacterially expressed human UCP3, reconstituted into liposomes, catalyses a strict exchange of aspartate, malate, sulphate and phosphate. The R282Q mutation abolishes the transport activity of the protein. Although the substrate specificity and inhibitor sensitivity of UCP3 display similarity with that of its close homolog UCP2, the two proteins significantly differ in their transport mode and kinetic constants.

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The androgen receptor (AR) is central to prostate cancer pathogenesis and has been extensively validated as a drug target. However, small-molecule anti-androgen therapies remain limited due to resistance and will eventually fail to suppress tumor growth, resulting in progression to castration-resistant prostate cancer (CRPC). The intrinsically disordered N-terminal domain (NTD) is crucial for AR transactivation and has been investigated as a suitable target in the presence of ligand binding domain mutations. A screening campaign identified biaryl isoxazole compound 7 as a weak inhibitor of the AR NTD. A library of biaryl analogues were synthesized, and their biological activities were assessed in a VCaP cell-based luciferase reporter gene assay. A structure-activity relationship (SAR) study revealed that indazole analogue 16 exhibited increased potency and favorable physicochemical properties with a benchmarked pharmacokinetic profile, providing a suitable starting point for further optimization of 16 as a CRPC therapeutic in the presence of AR mutations.

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There is an increased understanding of shared human-animal risk in terms of "one welfare", whereby when animals are at risk, so are people, so preventing injury to one species may also prevent injury to the other. Because transport-related injuries to horses are common, the authors considered this paradigm to study road equine transport-related injuries to humans in New Zealand. The aim was to determine their frequency and associated factors by distributing a survey to horse industry participants through industry organisations asking about their horse activities, road transport experiences, and any related self-injury. There were 112/1067 (10.5%) handlers injured while preparing (13/112), loading (39/112), traveling (6/112), or unloading (33/112). Of these, 40% had multiple injury types, and 33% had several body regions affected. Hand injuries were most common (46%), followed by the foot (25%), arm (17%), and head or face (15%). Median recovery time was 7 days. Injuries were associated with the responder's industry educational background, years of driving experience, and reporting a horse injured during road transport in the past two years. Wearing helmets and gloves, and adopting strategies designed to eliminate equine injuries associated with the road transport of horses to reduce the risk of injury to their handlers are recommended.

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The androgen receptor (AR) has been shown to be a key determinant in the pathogenesis of castration-resistant prostate cancer (CRPC). The current standard of care therapies targets the ligand-binding domain of the receptor and can afford improvements to life expectancy often only in the order of months before resistance occurs. Emerging preclinical and clinical compounds that inhibit receptor activity via differentiated mechanisms of action which are orthogonal to current antiandrogens show promise for overcoming treatment resistance. In this review, we present an authoritative summary of molecules that noncompetitively target the AR. Emerging small molecule strategies for targeting alternative domains of the AR represent a promising area of research that shows significant potential for future therapies. The overall quality of lead candidates in the area of noncompetitive AR inhibition is discussed, and it identifies the key chemotypes and associated properties which are likely to be, or are currently, positioned to be first in human applications.

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Biomarkers for osteoarthritis (OA) in horses have been extensively investigated, but translation into clinical use has been limited due to cost, limited sensitivity, and practicality. Identifying novel biomarkers that overcome these limitations could facilitate early diagnosis and therapy. This study aimed to compare the concentrations of synovial fluid (SF) and plasma cell-free DNA (cfDNA) over time in control horses with those with induced carpal OA. Following an established model, unilateral carpal OA was induced in 9 of 17 healthy Thoroughbred fillies, while the remainder were sham-operated controls. Synovial fluid and plasma samples were obtained before induction of OA (Day 0) and weekly thereafter until Day 63, and cfDNA concentrations were determined using fluorometry. The SF cfDNA concentrations were significantly higher for OA joints than for sham-operated joints on Days 28 (median 1430 µg/L and 631 µg/L, respectively, p = 0.017) and 63 (median 1537 µg/L and 606 µg/L, respectively, p = 0.021). There were no significant differences in plasma cfDNA between the OA and the sham groups after induction of carpal OA. Plasma cfDNA measurement is not sufficiently sensitive for diagnostic purposes in this induced model of OA. Synovial fluid cfDNA measurement may be used as a biomarker to monitor early disease progression in horses with OA.

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There is a lack of information on the number of horses shipped globally by air annually, the purpose of air travel and the routes of their journeys. This pilot study aimed to collect retrospective data on the international movements of horses by air from 2018 to 2021, describe their routes, and identify the possible effects of the coronavirus SARS-CoV-2 (COVID-19) pandemic. Equine transport data was gathered from 7 of 15 international shipping companies (ISCs) and 5 of 8 airlines contacted by email. The seven ISCs performed a median of 10,401 horse movements annually, ranging from a few hundred to several thousand movements per company, most frequently in Europe (Western and Northern Europe), Middle East/Africa (Middle East, Southern Africa), Asia Pacific (Australia), and the Americas (North and South America). The five airlines performed a median of 10,656 horse movements annually, importing and exporting horses to and from Europe, North America, Australasia, and the Middle East. For all but one airline, the number of horse movements decreased in 2020. The number and journey characteristics of horses transported by air require further scientific studies focused on the epidemiological and welfare risks unique to this type of transport to enable the development and implementation of best practices and regulations based on objective evidence.

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Brown adipose tissue (BAT) regulates metabolic physiology. However, nearly all mechanistic studies of BAT protein function occur in a single inbred mouse strain, which has limited the understanding of generalizable mechanisms of BAT regulation over physiology. Here, we perform deep quantitative proteomics of BAT across a cohort of 163 genetically defined diversity outbred mice, a model that parallels the genetic and phenotypic variation found in humans. We leverage this diversity to define the functional architecture of the outbred BAT proteome, comprising 10,479 proteins. We assign co-operative functions to 2,578 proteins, enabling systematic discovery of regulators of BAT. We also identify 638 proteins that correlate with protection from, or sensitivity to, at least one parameter of metabolic disease. We use these findings to uncover SFXN5, LETMD1, and ATP1A2 as modulators of BAT thermogenesis or adiposity, and provide OPABAT as a resource for understanding the conserved mechanisms of BAT regulation over metabolic physiology.

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Negative outcomes associated with the road transport of horses are a significant welfare issue. This study aimed to describe the injuries sustained by horses during road transport in New Zealand and factors associated with trauma while in transit. New Zealand horse industry participants were surveyed on their horse transport experiences and equine industry involvement. Participants were solicited through horse organisations. The data were tabulated, and a logistic regression was performed to identify significant (p < 0.05) factors associated with transport-related injury. In total, 201/1133 (17.7%/2 years) eligible surveys reported at least one horse injured during road transport. Most incidents occurred in transit (137/169; 81%), or when transported with one (76/193; 39.4%) or more (41/193; 21.2%) other horses. Most commonly, the hindlimbs, the head, or the forelimbs were injured (59.1%; 110/186 horses), ranging in severity from bruises to catastrophic orthopaedic trauma necessitating euthanasia. Eventing, not always checking horses' fitness for transport, using a tail guard or bandage, a stallion guard in the vehicle, bedding type on the floor, and behavioural problems were associated with injuries. This survey identified a significant incidence of injury and related death when horses are transported by road in New Zealand, and the key risk factors associated with the odds of injury.

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Uncoupling proteins (UCPs) form a distinct subfamily of the mitochondrial carrier family (MCF) SLC25. Four UCPs, DmUCP4A-C and DmUCP5, have been identified in Drosophila melanogaster on the basis of their sequence homology with mammalian UCP4 and UCP5. In a Parkinson's disease model, DmUCP4A showed a protective role against mitochondrial dysfunction, by increasing mitochondrial membrane potential and ATP synthesis. To date, DmUCP4A is still an orphan of a biochemical function, although its possible involvement in mitochondrial uncoupling has been ruled out. Here, we show that DmUCP4A expressed in bacteria and reconstituted in phospholipid vesicles catalyzes a unidirectional transport of aspartate, which is saturable and inhibited by mercurials and other mitochondrial carrier inhibitors to various degrees. Swelling experiments carried out in yeast mitochondria have demonstrated that the unidirectional transport of aspartate catalyzed by DmUCP4 is not proton-coupled. The biochemical function of DmUCP4A has been further confirmed in a yeast cell model, in which growth has required an efflux of aspartate from mitochondria. Notably, DmUCP4A is the first UCP4 homolog from any species to be biochemically characterized. In Drosophila melanogaster, DmUCP4A could be involved in the transport of aspartate from mitochondria to the cytosol, in which it could be used for protein and nucleotide synthesis, as well as in the biosynthesis of ß-alanine and N-acetylaspartate, which play key roles in signal transmission in the central nervous system.

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Aspartate has a central role in cancer cell metabolism. Aspartate cytosolic availability is crucial for protein and nucleotide biosynthesis as well as for redox homeostasis. Since tumor cells display poor aspartate uptake from the external environment, most of the cellular pool of aspartate derives from mitochondrial catabolism of glutamine. At least four transporters are involved in this metabolic pathway: the glutamine (SLC1A5_var), the aspartate/glutamate (AGC), the aspartate/phosphate (uncoupling protein 2, UCP2), and the glutamate (GC) carriers, the last three belonging to the mitochondrial carrier family (MCF). The loss of one of these transporters causes a paucity of cytosolic aspartate and an arrest of cell proliferation in many different cancer types. The aim of this review is to clarify why different cancers have varying dependencies on metabolite transporters to support cytosolic glutamine-derived aspartate availability. Dissecting the precise metabolic routes that glutamine undergoes in specific tumor types is of upmost importance as it promises to unveil the best metabolic target for therapeutic intervention.

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BACKGROUND: Mesenchymal stromal cells (MSCs) are believed to be hypoimmunogeneic with potential use for allogeneic administration. METHODS: Bone marrow was harvested from Connemara (n = 1), Standardbred (n = 6), and Thoroughbred (n = 3) horses. MSCs were grouped by their level of expression of major histocompatibility factor II (MHC II). MSCs were then sub-grouped by those MSCs derived from universal blood donor horses. MSCs were isolated and cultured using media containing fetal bovine serum until adequate numbers were acquired. The MSCs were cultured in xenogen-free media for 48 h prior to use and during all assays. Autologous and allogeneic MSCs were then directly co-cultured with responder leukocytes from the Connemara horse in varying concentrations of MSCs to leukocytes (1:1, 1:10, and 1:100). MSCs were also cultured with complement present and heat-inactivated complement to determine whether complement alone would decrease MSC viability. MSCs underwent haplotyping of their equine leukocyte antigen (ELA) to determine whether the MHC factors were matched or mismatched between the donor MSCs and the responder leukocytes. RESULTS: All allogeneic MSCs were found to be ELA mismatched with the responder leukocytes. MHC II-low and universal blood donor MSCs caused no peripheral blood mononuclear cell (PBMC) proliferation, no increase in B cells, and no activation of CD8 lymphocytes. Universal blood donor MSCs stimulated a significant increase in the number of T regulatory cells. Neutrophil interaction with MSCs showed that universal blood donor and MHC II-high allogeneic MSCs at the 6 h time point in co-culture caused greater neutrophil activation than the other co-culture groups. Complement-mediated cytotoxicity did not consistently cause MSC death in cultures with active complement as compared to those with inactivated complement. Gene expression assays revealed that the universal blood donor group and the MHC II-low MSCs were more metabolically active both in the anabolic and catabolic gene categories when cultured with allogeneic lymphocytes as compared to the other co-cultures. These upregulated genes included CD59, FGF-2, HGF, IDO, IL-10, IL-RA, IL-2, SOX2, TGF-ß1, ADAMSTS-4, ADAMSTS-5, CCL2, CXCLB/IL-8, IFNγ, IL-1ß, and TNFα. CONCLUSIONS: MHC II-low MSCs are the most appropriate type of allogeneic MSC to prevent activation of the innate and cell-mediated component of the adaptive immune systems and have increased gene expression as compared to other allogeneic MSCs.

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Adaptive thermogenesis has attracted much attention because of its ability to increase systemic energy expenditure and to counter obesity and diabetes1-3. Recent data have indicated that thermogenic fat cells use creatine to stimulate futile substrate cycling, dissipating chemical energy as heat4,5. This model was based on the super-stoichiometric relationship between the amount of creatine added to mitochondria and the quantity of oxygen consumed. Here we provide direct evidence for the molecular basis of this futile creatine cycling activity in mice. Thermogenic fat cells have robust phosphocreatine phosphatase activity, which is attributed to tissue-nonspecific alkaline phosphatase (TNAP). TNAP hydrolyses phosphocreatine to initiate a futile cycle of creatine dephosphorylation and phosphorylation. Unlike in other cells, TNAP in thermogenic fat cells is localized to the mitochondria, where futile creatine cycling occurs. TNAP expression is powerfully induced when mice are exposed to cold conditions, and its inhibition in isolated mitochondria leads to a loss of futile creatine cycling. In addition, genetic ablation of TNAP in adipocytes reduces whole-body energy expenditure and leads to rapid-onset obesity in mice, with no change in movement or feeding behaviour. These data illustrate the critical role of TNAP as a phosphocreatine phosphatase in the futile creatine cycle.

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OBJECTIVE: To evaluate thermal nociceptive thresholds (TNTs) before and after inducing a standardized radiocarpal bone osteochondral fracture (OCF) in horses. STUDY DESIGN: Prospective, controlled, randomized, masked study. ANIMALS: A group of 10 Thoroughbred fillies aged 2 years. METHODS: Skin temperature and TNTs were measured on the skin over the triceps brachii muscle in both the thoracic limbs before (week 0) and weekly (weeks 1-8) after unilateral arthroscopic induction of a radiocarpal OCF (n = 4) or sham surgery (n = 6) followed by a standardized exercise programme. The contralateral, non-operated thoracic limb was used as a control within each horse. Percentage thermal excursion (%TE) defined as %TE = 100 ∗ (TNT - skin temperature)/(cut-off temperature - skin temperature) was calculated. Data were analysed with a mixed-effects model followed by Dunnett's and Tukey's tests for within and between-limbs comparisons, respectively; p < 0.05 was considered significant. RESULTS: Skin temperature in the control limb of OCF horses was significantly higher at week 7 than at week 0 (p = 0.0125). At week 1, TNTs and %TE values in operated limbs of OCF horses were significantly reduced compared with their baseline values at week 0 (p ≤ 0.0153) and their values in contralateral control limbs (p ≤ 0.0024) and operated limbs of sham-operated horses (p ≤ 0.0162). At week 2, TNTs and %TE values in operated limbs of OCF horses remained significantly reduced compared with values in operated limbs of sham-operated horses (p ≤ 0.0248). CONCLUSIONS AND CLINICAL RELEVANCE: Creation of an OCF in a radiocarpal bone induced transitory (<2 weeks) ipsilateral heat hypersensitivity proximal to the surgery site (skin over the triceps brachii muscle) in horses. Surgically induced OCF may cause somatosensory abnormalities consistent with secondary thermal hyperalgesia.

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