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Topic:Experimental Methods

Experimental methods in equine research encompass a variety of techniques and procedures used to investigate physiological, behavioral, and health-related aspects of horses. These methods are designed to collect data that can enhance the understanding of equine biology and inform veterinary practices. Common experimental approaches include controlled trials, observational studies, and laboratory-based experiments. Techniques may involve the use of non-invasive imaging, biochemical assays, and physiological monitoring to assess parameters such as metabolic function, locomotion, and disease progression. This page compiles peer-reviewed research studies and scholarly articles that explore the application, development, and evaluation of experimental methods in the study of equine science.
Beyond the surface: how ex-vivo diffusion-weighted imaging reveals large animal brain microstructure and connectivity.
Frontiers in neuroscience    June 26, 2024   Volume 18 1411982 doi: 10.3389/fnins.2024.1411982
Behroozi M, Graïc JM, Gerussi T.Diffusion-weighted Imaging (DWI) is an effective and state-of-the-art neuroimaging method that non-invasively reveals the microstructure and connectivity of tissues. Recently, novel applications of the DWI technique in studying large brains through imaging enabled researchers to gain insights into the complex neural architecture in different species such as those of (e.g., horses and rhinos), (e.g., bovids, swines, and cetaceans), and (e.g., felids, canids, and pinnipeds). Classical tract-tracing methods are usually considered unsuitable for ethical and practical reasons, in large animals...
Oxytocin homogenizes horse group organization.
iScience    June 24, 2024   Volume 27, Issue 7 110356 doi: 10.1016/j.isci.2024.110356
Brooks J, Maeda T, Ringhofer M, Yamamoto S.The oxytocinergic system has been suggested to make up an important part of the endocrine basis of group cohesion. However, controlled studies in open-group settings have not been performed. We here investigated the impact of exogenous intranasal oxytocin on the group-level social organization of 5 groups of horses ( = 58; 12 mares and 46 geldings) through GPS tracking and social network analysis. We find oxytocin flattened social differentiation across levels. Most strikingly, oxytocin did not simply reinforce existing bonds but selectively shifted social preferences toward homogenization -...
Evaluation of pharmacokinetics of metoclopramide administered via subcutaneous bolus and intravenous constant rate infusion to adult horses.
Veterinary surgery : VS    June 24, 2024   doi: 10.1111/vsu.14128
Brandon AM, Williams JM, Davis JL, Martin EG, Capper AM, Crabtree NE.To determine the pharmacokinetics (PK) of metoclopramide administered via intravenous continuous rate infusion (IV CRI) and subcutaneous (SC) bolus and evaluate for gastrointestinal motility and adverse side effects. Methods: Experimental study; randomized, crossover design. Methods: Six healthy adult horses. Methods: Each horse received metoclopramide via IV CRI (0.04 mg/kg/h for 24 h) and SC bolus (0.08 mg/kg once), with ≥1 week washout period between. Plasma was analyzed by UPLC-MS/MS. Compartmental modeling was used to determine PK parameters for each treatment; nonparametric sup...
The Effects of Supplementation of the Freezing Extender with Silymarin on the Quality Parameters of Frozen-Thawed Arabian Stallion Sperm: A Preliminary Evaluation.
Biopreservation and biobanking    June 21, 2024   Volume 23, Issue 2 99-108 doi: 10.1089/bio.2023.0131
Khorsand F, Hamali H, Qasemi-Panahi B, Tohidkia M.This study evaluated the effects of supplementation of the freezing extender with different concentrations of silymarin on the quality of frozen-thawed Arabian stallion spermatozoa. Semen samples from three stallions (1, 2, and 3) were suspended in the freezing extender without or with silymarin (0, 25 μg/mL, 50 μg/mL, 75 μg/mL, and 100 μg/mL) and cryopreserved in 0.5 mL straws. After 1 month of storage, the frozen semen samples in straws were thawed and evaluated in terms of viability, mitochondrial membrane potential, kinematic parameters, total and progressive motility, plasma membrane ...
Viscoelastic properties of the equine hoof wall.
Acta biomaterialia    June 20, 2024   Volume 184 264-272 doi: 10.1016/j.actbio.2024.06.022
Bonney C, Pang S, Meyers MA, Jasiuk I.The equine hoof wall has outstanding impact resistance, which enables high-velocity gallop over hard terrain with minimum damage. To better understand its viscoelastic behavior, complex moduli were determined using two complementary techniques: conventional (∼5 mm length scale) and nano (∼1 µm length scale) dynamic mechanical analysis (DMA). The evolution of their magnitudes was measured for two hydration conditions: fully hydrated and ambient. The storage modulus of the ambient hoof wall was approximately 400 MPa in macro-scale experiments, decreasing to ∼250 MPa with hydration. In con...
Shape change in the saddle region of the equine back during trot and walk.
Journal of the Royal Society, Interface    June 19, 2024   Volume 21, Issue 215 20230644 doi: 10.1098/rsif.2023.0644
Smirnova KP, Frill MA, Warner SE, Cheney JA.Equine back pain is prevalent among ridden horses and is often attributed to poor saddle fit. An alternative explanation is that saddle fits are technically good but fit to the wrong configuration. Saddles are fit for the standing horse, but much of the time ridden is instead spent locomoting when the back experiences the greatest peak forces. We used an array of cameras to reconstruct the surface of the back and its movement during trot, walk and standing for five horses. We verified the setup's accuracy by reconstructing a laser-scanned life-sized model horse. Our reconstructions demonstrate...
The systemic cellular immune response against allogeneic mesenchymal stem cells is influenced by inflammation, differentiation and MHC compatibility: in vivo study in the horse.
Frontiers in veterinary science    June 18, 2024   Volume 11 1391872 doi: 10.3389/fvets.2024.1391872
Cequier A, Vázquez FJ, Vitoria A, Bernad E, Fuente S, Serrano MB, Zaragoza MP, Romero A, Rodellar C, Barrachina L.The effectiveness and safety of allogeneic mesenchymal stem/stromal cells (MSCs) can be affected by patient's immune recognition. Thus, MSC immunogenicity and their immunomodulatory properties are crucial aspects for therapy. Immune responses after allogeneic MSC administration have been reported in different species, including equine. Interactions of allogenic MSCs with the recipient's immune system can be influenced by factors like matching or mismatching for the major histocompatibility complex (MHC) between donor-recipient, and by the levels of MHC expression in MSCs. The latter can vary u...
Do tight nosebands have an effect on the upper airways of horses?
Veterinary medicine and science    June 17, 2024   Volume 10, Issue 4 e1478 doi: 10.1002/vms3.1478
Scholler D, Wittenberg J, Zablotski Y, May A.The public perception relating to the welfare of horses involved with equestrian sports is associated with training methods used and the presentation of horses at events. In this context, very tight nosebands, which are intended to prevent the horse from opening its mouth, also attract a lot of attention. Various studies have evaluated the impact of tight nosebands on stress parameters, whereas the effect of tight nosebands on upper airway function is unknown. Therefore, the aim of the study was to use overground endoscopy to evaluate changes in pharyngeal and laryngeal function when a tight n...
Bibliometric Analysis on Equine-Assisted Interventions.
Animals : an open access journal from MDPI    June 13, 2024   Volume 14, Issue 12 1776 doi: 10.3390/ani14121776
Amado-Fuentes M, Denche-Zamorano A, Barrios-Fernandez S, Gozalo M.Equine Assisted Interventions (EAIs) integrate the active participation of horses in therapeutic or educational interventions. A bibliometric analysis was carried out on this topic, using traditional bibliometric laws and recommendations. For this purpose, a search on the Web of Science (WoS) Core Collection database was carried out, obtaining 333 documents. Annual publications followed an exponentially increasing trend (R = 86%), pointing out that this topic is a growing interest among researchers, publishers, and journals. The USA was the most productive country worldwide and Jeong-yi Kwon a...
Induction of in vivo-like ciliation in confluent monolayers of re-differentiated equine oviduct epithelial cells.
Biology of reproduction    June 7, 2024   ioae090 doi: 10.1093/biolre/ioae090
Leemans B, Gadella BM, Marchand JHEAM, Van Soom A, Stout TAE.We recently developed re-differentiated equine oviduct epithelial cell (REOEC) monolayers demonstrating various in vivo morphological characteristics, but lacking secondary ciliation. In this study, we evaluated the effects of fetal bovine serum, reproductive steroid hormones, Wnt- and Notch ligands and inhibitors, and different EOEC seeding densities, in both conventional wells and on microporous membranes, on EOEC morphology and, in particular, secondary ciliation. REOEC monolayers were assessed by confocal microscopy after combined staining of nuclei, cilia and the cytoskeleton. Only Wnt li...
Heart rate and heart rate variability in horses undergoing hot and cold shoeing.
PloS one    June 6, 2024   Volume 19, Issue 6 e0305031 doi: 10.1371/journal.pone.0305031
Huangsaksri O, Wonghanchao T, Sanigavatee K, Poochipakorn C, Chanda M.Heart rate variability (HRV) is a frequently used indicator of autonomic responses to various stimuli in horses. This study aimed to investigate HRV variables in horses undergoing cold (n = 25) or hot (n = 26) shoeing. Multiple HRV variables were measured and compared between horses undergoing cold and hot shoeing, including the time domain, frequency domain, and nonlinear variables pre-shoeing, during shoeing, and at 30-minute intervals for 120 minutes post-shoeing. The shoeing method interacted with time to change the HRV variables standard deviation of RR intervals (SDNN), root mean square ...
Optimizing the preparation of paraffin sections from stallion testes.
Histology and histopathology    June 6, 2024   18775 doi: 10.14670/HH-18-775
Asgenbaatar N, Yi M, Wang X, Ulaangerel T, Shen Y, Wen X, Du M, Dong X, Mengkh Y, Dugarjav M, Bou G.The preparation of paraffin sections is an important experimental technique in animal histological research, and key factors that determine the quality of a section include the dehydration time, waxing time, and drying temperature of the paraffin section. Paraffin sections obtained from testis tissue of adult horses exhibited higher quality with clear tissue structure and complete cell morphology after they underwent gradient dehydration for 6 hours, were immersed in wax for 60 minutes, and were dried in a 75-degree oven for 15 minutes. The detailed, optimized procedures that are developed in ...
Monoclonal antibodies for equine CD25 improve detection of regulatory T cells in horses.
Veterinary immunology and immunopathology    June 6, 2024   Volume 274 110790 doi: 10.1016/j.vetimm.2024.110790
Wagner B, Babasyan S, Wilford S, Robbin MG, de Mestre AM.CD25, the interleukin-2 receptor α-chain, is expressed on cell surfaces of different immune cells and is commonly used for phenotyping of regulatory T cells (Tregs). CD25 has essential roles in the maintenance of hemostasis and immune tolerance and Treg cell involvement has been shown in human diseases and murine models for allergy, autoimmunity, cancer, chronic inflammation, and many others. In horses, a cross-reactive anti-human CD25 antibody has previously been used for characterizing Tregs. Here, we developed monoclonal antibodies (mAbs) to equine CD25 and compared their staining pattern ...
Flow-controlled expiration ventilation using a piston ventilator: effects of expiration time and speed on respiratory and pulmonary mechanics with focus on hysteresis and compliance in healthy horses.
American journal of veterinary research    June 5, 2024   Volume 85, Issue 8 ajvr.24.02.0036 doi: 10.2460/ajvr.24.02.0036
Douglas HF, Brandly J, Hopster K.To investigate the effects of FLow-controlled EXpiration (FLEX) ventilation expiration time and speed on respiratory and pulmonary mechanics in anesthetized horses in dorsal recumbency. Methods: 6 healthy adult research horses. Methods: In this randomized crossover experimental study, horses were anesthetized 3 times and were ventilated each time for 60 minutes using conventional volume-controlled ventilation (VCV), linear emptying of the lung over 50% of the expiratory time (FLEX50), or linear emptying of the lung over 100% of the expiratory time (FLEX100) in a randomized order. The primary o...
Equine Endothelial Cells Show Pro-Angiogenic Behaviours in Response to Fibroblast Growth Factor 2 but Not Vascular Endothelial Growth Factor A.
International journal of molecular sciences    May 30, 2024   Volume 25, Issue 11 6017 doi: 10.3390/ijms25116017
Finding EJT, Faulkner A, Nash L, Wheeler-Jones CPD.Understanding the factors which control endothelial cell (EC) function and angiogenesis is crucial for developing the horse as a disease model, but equine ECs remain poorly studied. In this study, we have optimised methods for the isolation and culture of equine aortic endothelial cells (EAoECs) and characterised their angiogenic functions in vitro. Mechanical dissociation, followed by magnetic purification using an anti-VE-cadherin antibody, resulted in EC-enriched cultures suitable for further study. Fibroblast growth factor 2 (FGF2) increased the EAoEC proliferation rate and stimulated scra...
Vascularity evaluation of the equine tarsocrural and proximal intertarsal joint septum, and comparative analysis of two arthroscopic transection techniques.
Veterinary surgery : VS    May 28, 2024   doi: 10.1111/vsu.14106
Labordère AL, Ruzickova P, Kamus LJ, Beauchamp G, Malo A, Richard H, Laverty S.To evaluate vascularity of the synovial membrane covered septum (SMS) separating the tarsocrural (TC) and proximal intertarsal (PIT) joints (Part 1) and compare two methods of transection, electrosurgical or Ferris Smith rongeur (FS rongeur) (Part 2). Methods: Experimental study. Methods: Part 1, 10 SMS (n = 5 horses). Part 2, six horses (n = 12 tarsi). Methods: In part 1, SMS harvested postmortem were each divided into eight regions of interest (ROIs), processed for histology, and immunostained with anti-α-actin antibody for blood vessel identification. Vascular density was calculate...
Transcutaneous Auricular Vagal Nerve Stimulation in Healthy Non-Sedated Horses: A Feasibility Study.
Veterinary sciences    May 28, 2024   Volume 11, Issue 6 241 doi: 10.3390/vetsci11060241
Vitale V, Bindi F, Velloso Alvarez A, de la Cuesta-Torrado M, Sala G, Sgorbini M.This study aimed to evaluate the feasibility of transcutaneous auricular vagal nerve stimulation (tAVNS) in healthy horses and its effect on heart rate variability (HRV). The study comprised three phases: the selection of mares, their acclimatization to the tAVNS, and the stimulation phase. Stimulation was performed with two electrodes positioned on the right pinna. The settings were 0.5 mA, 250 μs, and 25 Hz for pulse amplitude, pulse width, and pulse frequency, respectively. HRV was analysed before (B1), during (T), and after (B2) the tAVNS. From the 44 mares initially included, only 7 comp...
The Best Protocol to Treat Equine Skin Wounds by Second Intention Healing: A Scoping Review of the Literature.
Animals : an open access journal from MDPI    May 18, 2024   Volume 14, Issue 10 doi: 10.3390/ani14101500
Ribeiro G, Carvalho L, Borges J, Prazeres J.Equine skin wound treatment continues to be a challenge for veterinarians. Despite being a frequent practice, it remains difficult to choose an evidence-based treatment protocol. This study aimed to comprehensively explore the literature and provide a scoping review of therapeutic strategies for equine skin wounds and identify knowledge gaps and opportunities for future research. This review was conducted using specific criteria to select literature that described methods to manage second intention wound healing. After removing duplicates and screening papers for suitability, 81 manuscripts we...
The Poses for Equine Research Dataset (PFERD).
Scientific data    May 15, 2024   Volume 11, Issue 1 497 doi: 10.1038/s41597-024-03312-1
Li C, Mellbin Y, Krogager J, Polikovsky S, Holmberg M, Ghorbani N, Black MJ, Kjellström H, Zuffi S, Hernlund E.Studies of quadruped animal motion help us to identify diseases, understand behavior and unravel the mechanics behind gaits in animals. The horse is likely the best-studied animal in this aspect, but data capture is challenging and time-consuming. Computer vision techniques improve animal motion extraction, but the development relies on reference datasets, which are scarce, not open-access and often provide data from only a few anatomical landmarks. Addressing this data gap, we introduce PFERD, a video and 3D marker motion dataset from horses using a full-body set-up of densely placed over 100...
Expression of IL-10 and TGF-β1 in horses experimentally infected with T. equi merozoites is associated with antibody production but not modulation of pro-inflammatory responses.
Frontiers in immunology    May 13, 2024   Volume 15 1370255 doi: 10.3389/fimmu.2024.1370255
Onzere CK, Bastos RG, Bishop RP, Suarez CE, Fry LM. () is an apicomplexan parasite that causes severe hemolytic anemia in equids. Presently, there is inadequate knowledge of the immune responses induced by in equid hosts impeding understanding of the host parasite relationship and development of potent vaccines for control of infections. The objective of this study was to evaluate the host-parasite dynamics between merozoites and infected horses by assessing cytokine expression during primary and secondary parasite exposure, and to determine whether the pattern of expression correlated with clinical indicators of disease. Our findings showe...
Changes in characteristics of spermatogonial stem cells in response to heat stress in stallions.
Theriogenology    May 7, 2024   Volume 224 74-81 doi: 10.1016/j.theriogenology.2024.05.007
Shakeel M, Yoon M.Spermatogonial stem cells (SSCs) are essential for the maintenance of male fertility and survival of species. Environmental conditions, notably heat stress, have been identified as important causes of male infertility and have a negative impact on SSCs. Animals with cryptorchid testes (CT) are optimal models for the study of long-term heat stress-related changes in germ cells. The effect of heat stress on germ cells differs depending on the spermatogenesis stage. Thus, verifying whether the specific phase of spermatogenesis is dependent or independent of heat stress in stallions is important. ...
Effect of video angle on detection of induced front limb lameness in horses.
BMC veterinary research    May 3, 2024   Volume 20, Issue 1 172 doi: 10.1186/s12917-024-04032-9
Valle AP, Brown KA, Reilly P, Ciamillo SA, Davidson EJ, Stefanovski D, Stewart HL, Ortved KF.Lameness examinations are commonly performed in equine medicine. Advancements in digital technology have increased the use of video recordings for lameness assessment, however, standardization of ideal video angle is not available yielding videos of poor diagnostic quality. The objective of this study was to evaluate the effect of video angle on the subjective assessment of front limb lameness. A randomized, blinded, crossover study was performed. Six horses with and without mechanically induced forelimb solar pain were recorded using 9 video angles including horses trotting directly away and ...
Adaptive forgetting of place/object memory for dung in the domestic horse (Equus ferus caballus): Memory for a day.
Behavioural processes    March 24, 2024   Volume 217 105025 doi: 10.1016/j.beproc.2024.105025
Guyonnet AE, Whishaw IQ.The domestic horse (Equus ferus caballus) makes dung deposits to form "stud-piles" and compulsively examines dung droppings, suggesting that dung contains species-relevant information. The present study investigates horses' use of location (place), odor (object) and memory for dung sniff encounters. Horses were video recorded in 2 indoor and 4 outdoor riding arenas as they were taken at different time intervals to experimenter-determined objects or dung deposits that they could sniff. Frame-by-frame video analysis measured approaches, sniff duration, nostril use, ear position and blinking asso...
The impact of skin preparation method on electrocardiogram quality in horses.
The Canadian veterinary journal = La revue veterinaire canadienne    March 4, 2024   Volume 65, Issue 3 245-249 
McCrae P, Spong H, Mahnam A, Bashura Y, Pearson W.Several skin preparation techniques are used in electrocardiogram (ECG) monitoring of horses. The objective of this study was to determine which methods produce the greatest signal quality using textile electrodes and standard silver/silver chloride (Ag/AgCl) electrodes. Electrocardiogram data were collected using textile and Ag/AgCl electrodes simultaneously for 4 skin preparation techniques in 6 horses. The effects of skin preparation (cleansing with isopropyl alcohol, with or without shaving the hair) and the effects of the application of a conductive gel were assessed using metrics of sign...
Identification of the Reference Genes for Relative qRT-PCR Assay in Two Experimental Models of Rabbit and Horse Subcutaneous ASCs.
International journal of molecular sciences    February 14, 2024   Volume 25, Issue 4 2292 doi: 10.3390/ijms25042292
Ivanova Z, Petrova V, Grigorova N, Vachkova E.Obtaining accurate and reliable gene expression results in real-time RT-PCR (qRT-PCR) data analysis requires appropriate normalization by carefully selected reference genes, either a single or a combination of multiple housekeeping genes (HKGs). The optimal reference gene/s for normalization should demonstrate stable expression across varying conditions to diminish potential influences on the results. Despite the extensive database available, research data are lacking regarding the most appropriate HKGs for qRT-PCR data analysis in rabbit and horse adipose-derived stem cells (ASCs). Therefore,...
Evaluation of stability and safety of equine mesenchymal stem cells derived from amniotic fluid for clinical application.
Frontiers in veterinary science    February 14, 2024   Volume 11 1330009 doi: 10.3389/fvets.2024.1330009
Kim EY, Lee EJ, Kim RE, Kil TY, Kim MK.Amniotic fluid mesenchymal stem cells (AF-MSCs), which can be obtained from fetal tissue, reportedly have self-renewal capacity and multi-lineage differentiation potential. The aim of this study was to identify the biological characteristics of AF-MSCs and evaluate their stability and safety in long-term culture. To confirm the biological characteristics of AF-MSCs, morphology, proliferation capacity, karyotype, differentiation capacity, gene expression level, and immunophenotype were analyzed after isolating AF-MSCs from equine amniotic fluid. AF-MSCs were differentiated into adipocytes, chon...
A Relaxed Horse-A Relaxed Client? An Experimental Investigation of the Effects of Therapy Horses’ Stress on Clients’ Stress, Mood, and Anxiety.
Animals : an open access journal from MDPI    February 13, 2024   Volume 14, Issue 4 604 doi: 10.3390/ani14040604
Müller-Klein A, Braun MN, Ferreira de Sá DS, Michael T, Link-Dorner U, Lass-Hennemann J.Equine-assisted therapies are becoming increasingly popular for addressing physical and psychological disabilities in clients. The role of the horse's welfare in equine-assisted service receives increasing attention in research. Several studies have shown that horses are able to perceive human emotions and respond to human stress responses. However, no research has yet looked at the other side of the coin-whether and how humans perceive and react to equine stress levels during equine-assisted services. To fill this gap in the research, we employed a within-subjects design, in which horse-naïv...
The Behavioral Cost of Care: Changes in Maintenance Behavior during Equine-Assisted Interventions.
Animals : an open access journal from MDPI    February 6, 2024   Volume 14, Issue 4 536 doi: 10.3390/ani14040536
Fournier AK, French M, Letson EA, Hanson J, Berry TD, Cronin S.This study examined human-animal symbiosis in an animal-assisted intervention through observations of animal maintenance behaviors. The rise of psychotherapy, learning, and recreation incorporating animals warrants exploration of the welfare of the animals involved in these interventions. The analysis of welfare in multispecies engagements can be discussed in terms of symbiosis. Regarding an intervention's animal provider (e.g., therapy horse) and human recipient (psychotherapy client), the balance of cost and benefit is important. Research describing human and animal during interventions is ...
The ability of donkey sperm to induce oocyte activation and mule embryo development after ICSI.
Theriogenology    February 2, 2024   Volume 218 200-207 doi: 10.1016/j.theriogenology.2024.02.002
Arroyo-Salvo C, Cogollo Villarreal MY, Clérico G, Flores Bragulat AP, Niño Vargas A, Castañeira C, Briski O, Alonso C, Plaza J, Zeledon JM....Members of the Equus genus exhibit a fascinating capacity for hybridization, giving rise to healthy offspring. Mules, resulting from the mating of a mare with a jack, represent the most prevalent equid hybrid, serving diverse roles in our society. While in vitro embryo production, particularly through Intracytoplasmic Sperm Injection (ICSI), has rapidly gained significance in domestic horses, the in vitro production in other equids remains largely unexplored. Utilizing donkey sperm for fertilizing horse oocytes not only addresses this gap but also provides an opportunity to investigate donkey ...
Cell-permeable bone morphogenetic protein 2 facilitates bone regeneration by promoting osteogenesis.
Materials today. Bio    February 1, 2024   Volume 25 100983 doi: 10.1016/j.mtbio.2024.100983
Kang M, Lee S, Seo JP, Lee EB, Ahn D, Shin J, Paik YK, Jo D.The use of the FDA-approved osteoinductive growth factor BMP2 is widespread for bone regeneration. However, its clinical application has been hindered by limitations in cell permeability and a short half-life in circulation. To address this issue, we have developed a modified version of BMP2, referred to as Cell Permeable (CP)-BMP2, which possesses improved cell permeability. CP-BMP2 incorporates an advanced macromolecular transduction domain (aMTD) to facilitate transfer across the plasma membrane, a solubilization domain, and recombinant human BMP2. Compared to traditional rhBMP2, CP-BMP2 ex...
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