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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.
Study of the duration and distribution of equine influenza virus subtype 2 (H3N8) antigens in experimentally infected ponies in vivo. Sutton GA, Viel L, Carman PS, Boag BL.The purpose of this experiment was to study the duration and distribution of equine influenza virus in actively infected ponies over a 3 wk period. Pony foals (6-8 mo old) were infected experimentally by nebulizing equine influenza subtype-2 virus ultrasonically through a face mask. Successful infection was clinically apparent as each of the foals (n = 6) had a febrile response, a deep hacking cough and mucopurulent nasal discharge for 7 to 10 d. The virus was isolated from nasopharyngeal swabs of all the ponies 3 and 5 d after infection and all the ponies seroconverted to the virus. Samples w...
Use of Rhodococcus equi virulence-associated protein for immunization of foals against R equi pneumonia.
American journal of veterinary research    April 1, 1997   Volume 58, Issue 4 356-359 
Prescott JF, Nicholson VM, Patterson MC, Zandona Meleiro MC, Caterino de Araujo A, Yager JA, Holmes MA.To evaluate use of the virulence-associated protein of Rhodococcus equi in immunizing foals against R equi pneumonia. Methods: Eight (experimental group) and 6 (controls) mares with their foals. Methods: Virulence-associated protein extracted from R equi was used to prepare an acetone-precipitated. Triton X-extracted (APTX) antigen. After determination of the efficacy of passive immunization, in untreated foals or in foals given plasma from a horse vaccinated with APTX antigen or from a nonvaccinated horse, a field trial was done to evaluate the efficacy of vaccination of 8 mares, twice with A...
Corneal concentrations and preliminary toxicological evaluation of an itraconazole/dimethyl sulphoxide ophthalmic ointment.
Journal of veterinary pharmacology and therapeutics    April 1, 1997   Volume 20, Issue 2 100-104 doi: 10.1046/j.1365-2885.1997.00822.x
Ball MA, Rebhun WC, Trepanier L, Gaarder J, Schwark WS.The objectives of this study were to determine the concentration of itraconazole achieved in corneal tissue and aqueous humour after topical application of a 1% itraconazole ointment: to determine the effect of including dimethyl sulphoxide (DMSO) in the ointment on achievable ocular tissue itraconazole concentrations; and to assess if any gross or histopathologic ocular toxicity results from the topical application of 1% itraconazole with or without the addition of DMSO. The experimental trial consisted of 6 horses considered to have normal eyes. Each horse had one eye treated with 0.3 mL of ...
An in vitro biomechanical investigation of the mechanical properties of dynamic compression plated osteotomized adult equine tibiae.
Veterinary surgery : VS    March 1, 1997   Volume 26, Issue 2 126-136 doi: 10.1111/j.1532-950x.1997.tb01475.x
McD○ LA, Stover SM, Taylor KT.To determine the monotonic mechanical properties of osteotomized adult equine tibiae stabilized with two dynamic compression plates (DCP) and to compare the mechanical properties with those of intact tibiae and in vivo loads. Methods: The compressive, bending, and torsional mechanical properties of plated and intact tibiae were assessed in vitro. Methods: Twelve pairs of adult equine tibiae. Methods: Tibiae were loaded in axial compression, craniocaudal 3-point bending, or torsion in external rotation in a single cycle to failure. Mechanical properties were determined from load-displacement da...
Methods for the isolation, culture and characterisation of equine pulmonary artery endothelial cells.
Research in veterinary science    March 1, 1997   Volume 62, Issue 2 147-152 doi: 10.1016/s0034-5288(97)90137-5
MacEachern KE, Smith GL, Nolan AM.Equine endothelial cells were isolated from the pulmonary artery by enzymatic digestion and grown to confluency. The cells were characterised by positive immunofluorescent staining for von Willebrand factor and NADPH-diaphorase staining for nitric oxide synthase. Measurements of endothelins indicated that there were significant release rates from the cells for up to six hours. Measurements of intracellular calcium concentration showed that the application of bradykinin caused a transient increase in calcium concentration with similar characteristics to those observed in other endothelial cell ...
Determination of an effective dose of eltenac and its comparison with that of flunixin meglumine in horses after experimentally induced carpitis.
American journal of veterinary research    March 1, 1997   Volume 58, Issue 3 298-302 
Hamm D, Turchi P, Johnson JC, Lockwood PW, Thompson KC, Katz T.To titrate a clinically effective eltenac dosage (0.1, 0.5, and 1.0 mg/kg of body weight), compared with vehicle only, and to compare efficacy of the most effective eltenac dosage with that of 1.1 mg of flunixin meglumine/kg. Methods: 40 healthy horses, ranked after model induction on the basis of lameness severity, were randomly assigned to 5 treatment groups, with 4 replicates of 10 horses each. Methods: On day -5, after surgical preparation of the left carpal region, 0.7 ml of Freund's complete adjuvant was injected into the intercarpal space. Horses were observed daily, from the day of car...
Intra-articular anesthesia of the distal interphalangeal joint alleviates lameness associated with the navicular bursa in horses.
Veterinary surgery : VS    March 1, 1997   Volume 26, Issue 2 137-140 doi: 10.1111/j.1532-950x.1997.tb01476.x
Pleasant RS, Moll HD, Ley WB, Lessard P, Warnick LD.To determine if intra-articular anesthesia of the distal interphalangeal joint could alleviate lameness associated with the navicular bursa in horses. Methods: Experimental investigation. Methods: Six clinically normal horses. Methods: Lameness was induced in each horse by injecting either the left or right front navicular bursa with 5 mg of amphotericin-B. Forty-eight hours later each horse was videotaped walking and trotting before, and 5, 30, and 60 minutes after injecting the distal interphalangeal joint of the treated limb with 5 mL of 2% mepivacaine hydrochloride. All video recordings we...
Artifactual nonlinearity due to wear grooves and friction in four-point bending experiments of cortical bone.
Journal of biomechanics    February 1, 1997   Volume 30, Issue 2 185-188 doi: 10.1016/s0021-9290(96)00124-8
Griffin LV, Gibeling JC, Gibson VA, Martin RB, Stover SM.Experiments and analyses were performed to determine the cause of a nonlinear force-deflection response observed in four-point flexural fatigue of beams of cortical bone machined from the mid-diaphysis of the equine third metacarpus. Observable grooves which formed on the beam surface at supports and load noses were found to be the primary cause of the nonlinearity. An additional geometric nonlinearity at large deflections revealed by finite element modeling may be minimized by using the smallest diameter supports and load noses recommended in ASTM 790. However, frictional constraint of the be...
Experimental induction of equine protozoal myeloencephalitis in horses using Sarcocystis sp. sporocysts from the opossum (Didelphis virginiana).
Veterinary parasitology    February 1, 1997   Volume 68, Issue 3 199-213 doi: 10.1016/s0304-4017(96)01112-0
Fenger CK, Granstrom DE, Gajadhar AA, Williams NM, McCrillis SA, Stamper S, Langemeier JL, Dubey JP.Sarcocystis sp. sporocysts isolated from eight feral opossums (Didelphis virginiana) were pooled and fed to 18 commercially reared budgerigars (Melopsittacus undulatus), 14 wild-caught sparrows (Passer domesticus), one wild-caught slate-colored Junco (Junco hyemalis) and five weanling horses (Equus caballus). All budgerigars died within 5 weeks post inoculation (wpi). Histologic examination revealed meronts within the pulmonary epithelia and typical Sarcocystis falcatula sarcocysts developing in the leg muscles. Sparrows were euthanized 13 and 17 wpi and their carcasses were fed to four labora...
Serologic markers in early stages of African horse sickness virus infection.
Journal of clinical microbiology    February 1, 1997   Volume 35, Issue 2 531-535 doi: 10.1128/jcm.35.2.531-535.1997
Martínez-Torrecuadrada JL, Díaz-Laviada M, Roy P, Sánchez C, Vela C, Sánchez-Vizcaíno JM, Casal JI.Fifteen horses were experimentally infected with African horse sickness virus (AHSV) serotype 4. To learn more about the time course of production and specificity of AHSV-specific antibodies, sera were analyzed by immunoblot analysis. Only animals that survived for more than 9 days were able to develop a humoral immune response detectable by immunoblotting. The earliest serological markers corresponded mainly to VP5, VP6, and NS2 and to a lesser extent to VP3, NS1, and NS3. Neutralizing antibodies to VP2 were not detected by immunoblotting, suggesting that they are mostly conformation dependen...
Method for the growth of equine airway epithelial cells in culture.
Research in veterinary science    January 1, 1997   Volume 62, Issue 1 30-33 doi: 10.1016/s0034-5288(97)90176-4
Sime A, McKellar Q, Nolan A.A serum-free cell culture method was developed for equine tracheal epithelial cells which allowed the growth and characterisation of the phenotypical properties of this cell type. Several variables influenced the efficacy of the attachment and growth of the isolated cells. Serum and a collagen matrix were essential components for efficient cell attachment. Once attachment had occurred, cell growth was enhanced by a serum-free medium containing bovine pituitary extract, retinoic acid, insulin, hydrocortisone, transferrin, epidermal growth factor, adrenaline and triiodothyronine. The mean time t...
Effect of 19-norandrostenololylaurate on testicular growth in colts.
Acta veterinaria Scandinavica    January 1, 1997   Volume 38, Issue 1 51-57 doi: 10.1186/BF03548507
Koskinen E, Andersson M, Katila T.The effect of anabolic steroid on testicular growth was investigated in 3 experiments. In experiment I, 500 mg of the anabolic steroid was given to 4 colts and 100 mg to another 4 colts, every 3rd week, starting at age 16 months and ending at age 24 months. Six colts served as controls. Both treatments decreased total scrotal width (TSW) within 6 weeks. Seasonal testicular growth during spring partly overcame the effect of steroid treatment. Cessation of anabolic steroid treatment was followed by testicular growth at the same time as TSW in untreated colts was decreasing by virtue of the effec...
Local electrostatic potentials in pyridoxal phosphate labelled horse heart cytochrome c.
Journal of photochemistry and photobiology. B, Biology    January 1, 1997   Volume 37, Issue 1-2 74-83 doi: 10.1016/s1011-1344(96)07345-9
Miteva MA, Kossekova GP, Villoutreix BO, Atanasov BP.The present work shows the application of an optical label pyridoxal phosphate (PLP) for the experimental determination of local electrostatic potentials in singly substituted cytochromes c modified by pyridoxal phosphate at Lys 79 (PLP-Lys-79-cyt.c) or at Lys 86 (PLP-Lys-86-cyt.c). PLP has also been used to calculate the pKa values of all ionizable groups and the electrostatic potentials in the modified proteins and to analyse their properties. The experimental pKa values for the pyridine nitrogen and phenolic hydroxyl of the bound label were obtained from pH-dependent absorbance and fluoresc...
Alterations in equine guttural pouch morphology with head position: observations using a new technique for producing accurate casts.
The Anatomical record    December 1, 1996   Volume 246, Issue 4 579-584 doi: 10.1002/(SICI)1097-0185(199612)246:43.0.CO;2-L
Babptiste KE, Holladay SD, Freeman LE.An improved technique for preparing casts of the equine guttural pouch is described. The new technique is simplified and inexpensive in comparison to previous techniques and produces highly accurate anatomic models which are both durable and flexible. Methods: Silicone rubber bathtub caulk was injected into the guttural pouches of horse cadavers. The silicone was allowed to cure overnight, and casts were removed by dissection. The new technique was then used to evaluate changes in the shape of the guttural pouch with changes in head position. Results: With flexion of the atlanto-occipital join...
Anion secretion induced by capacitative Ca2+ entry through apical and basolateral membranes of cultured equine sweat gland epithelium.
The Journal of physiology    November 15, 1996   Volume 497 ( Pt 1), Issue Pt 1 19-29 doi: 10.1113/jphysiol.1996.sp021746
Ko WH, Chan HC, Wong PY.1. Anion secretion induced by capacitative Ca2+ entry through apical and basolateral membranes of cultured equine sweat gland epithelium was studied using the short-circuit current (Isc) technique. 2. Thapsigargin induced an increase in Isc that could be inhibited when external Ca2+ was chelated by EGTA. 3. The inhibition of the thapsigargin-induced Isc could be reversed by re-addition of Ca2+ to apical or basolateral solutions. The magnitude of the reactivated Isc depended predominantly on basolateral Ca2+ concentration. 4. The magnitude of the reactivated Isc upon basolateral Ca2+ addition i...
Efficacy of prosthetic laryngoplasty with and without bilateral ventriculocordectomy as treatments for laryngeal hemiplegia in horses.
American journal of veterinary research    November 1, 1996   Volume 57, Issue 11 1668-1673 
Tetens J, Derksen FJ, Stick JA, Lloyd JW, Robinson NE.To evaluate the efficacy of prosthetic laryngoplasty with and without bilateral ventriculocordectomy for treatment of experimentally induced left laryngeal hemiplegia (LLH). Methods: 15 adult Standardbreds. Methods: Horses were allotted to 3 equal groups. Sham operation (group 1), prosthetic laryngoplasty (group 2), or prosthetic laryngoplasty with bilateral ventriculocordectomy (group 3) was performed after induction of LLH. Upper airway function testing was performed prior to left recurrent laryngeal neurectomy (LRLN), 14 days after LRLN, and 60 and 180 days after surgical treatment. Measure...
Role of carbohydrates in the attachment of equine spermatozoa to uterine tubal (oviductal) epithelial cells in vitro.
American journal of veterinary research    November 1, 1996   Volume 57, Issue 11 1635-1639 
Dobrinski I, Ignotz GG, Thomas PG, Ball BA.To test the hypotheses that the attachment of equine spermatozoa to uterine tubal (oviductal) epithelial cells (OEC) in vitro is mediated by glycoproteins, and that proteins with carbohydrate-binding properties are present in the periacrosomal plasma membrane of equine spermatozoa. Methods: 4 reproductively sound stallions, and 1 mare in estrus. Methods: In experiment 1a, fluorescent-labeled spermatozoa were cocultured with monolayers of OEC in the presence of 50 mM glucose, fructose, galactose, mannose, N-acetyl glucosamine, N-acetyl galactosamine, or N-acetyl neuraminic acid, or 10 mg of fet...
Efficacy of an epidural combination of morphine and detomidine in alleviating experimentally induced hindlimb lameness in horses.
Veterinary surgery : VS    November 1, 1996   Volume 25, Issue 6 511-518 doi: 10.1111/j.1532-950x.1996.tb01452.x
Sysel AM, Pleasant RS, Jacobson JD, Moll HD, Modransky PD, Warnick LD, Sponenberg DP, Eyre P.Amphotericin B-induced synovitis of the left tarsocrural joint was used to create a grade 3 of 4 lameness in 11 horses. Caudal epidural catheters were placed and advanced to the lumbosacral region. Baseline heart and respiratory rates were recorded and horses were videotaped at a walk and trot. Morphine sulphate (0.2 mg/kg) and detomidine hydrochloride (30 micrograms/kg) were administered to treated horses (n = 8) through the epidural catheter; an equivalent volume of physiologic saline solution was administered to control horses (n = 3) through the catheter. At hourly intervals after epidural...
Experimental contact of bighorn sheep (Ovis canadensis) with horses and cattle, and comparison of neutrophil sensitivity to Pasteurella haemolytica cytotoxins.
Journal of wildlife diseases    October 1, 1996   Volume 32, Issue 4 594-602 doi: 10.7589/0090-3558-32.4.594
Foreyt WJ, Lagerquist JE.Peripheral blood neutrophils from horses, cattle, and Rocky Mountain bighorn sheep (Ovis canadensis canadensis) were evaluated for susceptibility to cytotoxin-dependent lysis of different biotypes and serotypes of Pasteurella haemolytica of domestic sheep, cattle, bighorn sheep, or mountain goat (Oreamnos americana) origin utilizing a cytotoxicity assay which measures the degree of bacteria cytotoxin-killing of neutrophils. All isolates of P. haemolytica (biotypes A and T) were noncytotoxic to horse neutrophils. Thirteen of 18 R haemolytica biotype A isolates were cytotoxic (> 50% neutrophi...
Maintenance of equine articular cartilage explants in serum-free and serum-supplemented media, compared with that in a commercial supplemented medium.
American journal of veterinary research    September 1, 1996   Volume 57, Issue 9 1261-1265 
Kawcak CE, Trotter GW, Frisbie DD, McIlwraith CW.To evaluate the effects of a commercially defined, serum-free medium additive on equine articular cartilage explants, compared with effects of serum-free and serum-supplemented media. Methods: Articular cartilage from a 3-year-old, mixed breed horse euthanatized for reasons other than musculoskeletal disease or sepsis. Methods: Media were changed every 48 hours, and the glycosaminoglycan (GAG) content was determined in media collected at each time point. Glycosaminoglycan synthesis by explant chondrocytes, and residual GAG content of articular cartilage (as a measure of explant GAG loss) were ...
Molecular diffusion into horse spleen ferritin: a nitroxide radical spin probe study.
Biophysical journal    September 1, 1996   Volume 71, Issue 3 1587-1595 doi: 10.1016/S0006-3495(96)79361-X
Yang X, Chasteen ND.Electron paramagnetic resonance spectroscopy and gel permeation chromatography were employed to study the molecular diffusion of a number of small nitroxide spin probes (approximately 7-9 A diameter) into the central cavity of the iron-storage protein ferritin. Charge and polarity of these radicals play a critical role in the diffusion process. The negatively charged radical 4-carboxy-2,2,6,6-tetramethylpiperidine-N-oxyl (4-carboxy-TEMPO) does not penetrate the cavity whereas the positively charged 4-amino-TEMPO and 3-(aminomethyl)-proxyl radical and polar 4-hydroxy-TEMPO radical do. Unlike th...
Experimental pathological studies on mechanism of abortion caused by equine arteritis virus.
The Japanese journal of veterinary research    August 1, 1996   Volume 44, Issue 2 133-135 
Wada R.No abstract available
Instrumentation and techniques in respiratory surgery.
The Veterinary clinics of North America. Equine practice    August 1, 1996   Volume 12, Issue 2 351-372 doi: 10.1016/s0749-0739(17)30286-9
Stick JA.In this article, videoendoscopy in conjunction with high-speed treadmill exercise and other specialized diagnostic techniques used to measure upper airway function are reviewed as they pertain to the four most commonly diagnosed functional upper airway obstructions: (1) laryngeal hemiplegia, (2) arytenoid chondritis, (3) epiglottic entrapment, and (4) intermittent dorsal displacement of the soft palate. Using experimental and clinical findings obtained with these new instruments and diagnostic techniques, current recommendations for surgery of these obstructions are presented.
Comparison of media and sera used for in vitro culture of equine superficial digital flexor tendon explants.
American journal of veterinary research    August 1, 1996   Volume 57, Issue 8 1118-1123 
Riley CB, Archer FJ, Bailey JV.To compare the effects of different commercial nutrient media and sera on protein synthesis and maintenance of cellular density in cultures of the equine superficial digital flexor tendon (SDFT). Methods: 8 healthy 2- to 4-year-old horses. Methods: First Dulbecco's modified Eagle's medium, Ham's F12 nutrient mixture, RPMI 1640 medium, minimum essential medium with Earle's salts, minimum essential medium with Hanks' salts, and a Dulbecco's modified Eagle's medium/Ham's F12 nutrient mixture with 10% fetal bovine serum (FBS) were compared. Then FBS, fetal equine serum, and donor horse serum, each...
Effect of methylprednisolone acetate on proteoglycan and collagen metabolism of articular cartilage explants.
The Journal of rheumatology    July 1, 1996   Volume 23, Issue 7 1207-1213 
Todhunter RJ, Fubini SL, Wootton JA, Lust G.The effect of different doses of methylprednisolone acetate (MPA) on proteoglycan and collagen metabolism of articular cartilage from normal equine joints was tested in vitro. Methods: Cultured explants were treated with 0, 0.0004, 0.004, 0.04, 0.4 and 4.0 mg/ml (approximately 10(-6)-10(-2) M) MPA for 72 h. Proteoglycan synthesis was measured by incorporation of sodium [35S]sulfate into proteoglycans and proteoglycan degradation was measured by release of total and radiolabeled proteoglycan into the culture media. The size of the proteoglycans was assessed with size exclusion chromatography un...
In vitro evaluation of four methods of attaching transfixation pins into a fiberglass cast for use in horses.
American journal of veterinary research    July 1, 1996   Volume 57, Issue 7 1098-1101 
McClure SR, Watkins JP, Hogan HA.To compare the axial stability provided by 4 methods of attaching transfixation pins into a fiberglass cast. Methods: Axial stability of 4 methods of transfixation pin attachment to a fiberglass cast cylinder was determined in vitro. Methods of attachment included simple incorporation of the pins into the cast, placement of a washer and nut on the pin and incorporation into the cast, extension of pins beyond the cast and attachment to a steel halo, and washers within the cast and attachment to a steel halo. Methods: A model was designed to simulate a transfixation cast applied to the equine me...
Experimental model for the study by chemiluminescence of the activation of isolated equine leucocytes.
Research in veterinary science    July 1, 1996   Volume 61, Issue 1 59-64 doi: 10.1016/s0034-5288(96)90112-5
Benbarek H, Deby-Dupont G, Deby C, Caudron I, Mathy-Hartert M, Lamy M, Serteyn D.The activation of human polymorphonuclear leucocytes (the respiratory burst) can be studied by measuring their chemiluminescent response. This technique was adapted to equine leucocytes to investigate the effects of cell number, activator concentration, enhancers of chemiluminescence, pH, temperature and inhibitors. Leucocytes were isolated from citrated blood from healthy horses and chemiluminescence was measured with a Bio-Orbit luminometer sensitive to 900 nm light. The optimal cell density for the maximal chemiluminescent response ranged from 10(6) to 10(7) leucocytes 600 microliters-1. Ch...
Ionic mechanisms of Ca(2+)-dependent electrolyte transport across equine sweat gland epithelium.
The Journal of physiology    June 15, 1996   Volume 493 ( Pt 3), Issue Pt 3 885-894 doi: 10.1113/jphysiol.1996.sp021431
Ko WH, Chan HC, Chew SB, Wong PY.1. The ionic mechanism involved in Ca(2+)-stimulated electrolyte transport in cultured equine sweat gland epithelial cells was studied using the short-circuit current (ISC) technique. 2. Microscopy revealed that the cultured cells grown on Millipore filters formed polarized monolayers with tight junctions. Monolayers exhibited a mean transepithelial resistance of 333.9 +/- 40.4 omega cm2. 3. Ca(2+)-mobilizing agents, A23187 (1 microM) or thapsigargin (0.01-1 microM), stimulated ISC while forskolin exerted little effect on the ISC. 4. Replacement of external Cl- by gluconate significantly reduc...
Influence of glycerol on the structure and stability of ferric horse heart myoglobin: a SAXS and circular dichroism study.
Biochimica et biophysica acta    June 7, 1996   Volume 1295, Issue 1 51-58 doi: 10.1016/0167-4838(96)00010-6
Barteri M, Gaudiano MC, Santucci R.The influence of glycerol on the structural properties of Fe(III)-horse heart myoglobin has been investigated by absorbance, CD and SR-SAXS spectroscopy. The results obtained indicate that both the tertiary and the secondary (alpha-helix) conformations of the protein are influenced by glycerol; in particular, an increase of approx. 8% in helical content was observed. Further, analysis of both the acid- and guanidine-induced denaturation transitions points to a glycerol-induced decreased stability of the tertiary structure; conversely, the alpha-helix conformation is found to be stabilized by t...
Use of adverse conditions to stimulate a cellular stress response by equine articular chondrocytes.
American journal of veterinary research    June 1, 1996   Volume 57, Issue 6 860-865 
Benton HP, Cheng TC, MacDonald MH.To determine the response of equine articular cartilage cells to heat and calcium stresses. Methods: Analysis of newly synthesized, [35S]methionine-labeled proteins after treatment of isolated primary equine chondrocytes. Methods: Primary cultures of equine articular chondrocytes were incubated at temperatures ranging from 37 to 42 C for heat stress experiments or incubated in the presence or absence of the intracellular calcium pump inhibitor, thapsigargin, for calcium stress experiments. Patterns of new protein synthesis were determined by incubating with [35S]methionine followed by separati...
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