In vitro research involving horses refers to the study of equine cells, tissues, or biological molecules outside their normal biological context, typically in controlled laboratory environments. This research approach allows scientists to investigate cellular processes, molecular interactions, and the effects of various treatments without the ethical and logistical complexities of in vivo studies. In vitro studies contribute to understanding equine physiology, pathology, and pharmacology by providing insights into cellular responses to pathogens, drugs, and other stimuli. This page compiles peer-reviewed research studies and scholarly articles that explore various in vitro methodologies and their applications in equine science, including cell culture techniques, molecular assays, and drug efficacy testing.
Gargouri Y, Pieroni G, Rivière C, Sugihara A, Sarda L, Verger R.We report further investigations on protein inhibition of pancreatic and microbial lipases carried out with the monolayer technique. When beta-lactoglobulin A, melittin, serum albumin, myoglobin, and a protein inhibiting lipase from soybean were preincubated with a dicaprin film at a surface pressure of 35 dynes/cm, no activity was detected with horse pancreatic or Rhizopus delemar lipases. By contrast, Rhizopus arrhizus and Geotrichum candidum lipase activities were not impaired under the same conditions. Experiments using mixed lipid-protein film transfer clearly show that the inhibition of ...
Freeman KP, Roszel JF, Slusher SH.Inclusions and cellular changes were seen in cytologic specimens from 1 healthy horse, 6 horses hospitalized because of respiratory problems, and 1 horse hospitalized because of colic and hepatitis. Two bronchial aspirates contained detached ciliated cytoplasmic tufts and cytoplasmic inclusions characteristic of the specific degenerative process called ciliocytophthoria. These changes and inclusions resembled those seen in bronchial aspirates from human beings with parainfluenza virus infection. Four bronchial aspirates and 2 serous fluid specimens had nuclear inclusions resembling those seen ...
Adamson PJ, Wilson WD, Hirsh DC, Baggot JD, Martin LD.In vitro antimicrobic susceptibility patterns of commonly isolated aerobic gram-positive and gram-negative bacterial pathogens of equine origin were determined, using the agar-plate dilution method. All organisms were recent clinical isolates and included Corynebacterium (Rhodococcus) equi, Corynebacterium pseudotuberculosis, (coagulase positive) Staphylococcus sp, Streptococcus equi, Streptococcus zooepidemicus, Actinobacillus sp, Enterobacter cloacae, Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa, and Salmonella. In vitro susceptibility levels were outlined for 14 antimicro...
Harrington JP, Carrier TL.The effect of several different types of perturbants and pH on the rate of autoxidation of horse heart ferrocytochrome c was investigated. The kinetic behavior is unique to each perturbant used. Rates of autoxidation followed first-order kinetics over the time span (0-180 min) studied. The Cl- and Br- anions exhibit an initial increase in the rate of autoxidation up to 100 mM, followed by a decrease in kinetics at 500 mM anion concentration. The ClO4- anion exhibits only an increase in the rate of autoxidation with increasing ionic strength, where as, propylurea, a hydrophobic perturbant, is n...
Ostlund C, Pero RW, Johnson DB.This study compares the relationship between N-acetoxy-2-acetylaminofluorene (NA-AAF) and u.v. induced unscheduled DNA synthesis (UDS) and their respective relationships to age and blood pressure in horse mononuclear leukocytes with earlier, similar investigations on human leukocytes. U.v. induced UDS was found to proceed more rapidly than NA-AAF induced UDS. A pronounced lag period associated with the rapid demand for 3H-dThd into DNA after u.v. damage was observed. NA-AAF induced UDS correlated significantly with NA-AAF binding, age and the blood pressure of male horses. UDS values, induced ...
Dubin A, Potempa J, Silberring J.Horse leucocyte neutral proteinase inhibitor reacts with all tested elastases at the molar ratios of 1:1 and yielding stable complexes (Ki = 10(-10) M). The above reactions are very rapid, characterized by the high values of association rate constant kon = 10(7) M-1s-1.
Aguado LI, Ojeda SR.During the days preceding the first ovulation the ovary of the rat exhibits a remarkable increase in estradiol (E2) and progesterone (P) release in response to gonadotropins. No such increase is observed in the case of androgens (A, testosterone + dihydrotestosterone). The present experiments were undertaken to examine the possibility of reproducing these developmental events by stimulating the ovary with a gonadotropin that has substantial FSH-like activity. In vivo administration of pregnant mare serum gonadotropin (PMSG) to juvenile 29-day-old rats greatly increased the in vitro E2 and A re...
Magnuson NS, Perryman LE, Suttle DP, Robinson JL, Mason PH, Marta KM.In an attempt to determine the metabolic defect causing severe combined immunodeficiency (SCID) in horses in which altered purine metabolism has been observed, various parameters of purine and pyrimidine metabolism were evaluated. The activities of nine purine enzymes (adenosine kinase, purine nucleoside phosphorylase, deoxyadenosine kinase, deoxycytidine kinase, 5'-nucleotidase, AMP deaminase, hypoxanthine-guanine phosphoribosyl transferase, and adenine phosphoribosyl transferase were measured in fibroblasts. All activities determined for SCID horses were normal. Uptake of 10 microM adenosine...
Matoba H, Allen JR, Bayly WM, Oakley CR, Gollnick PD.Comparisons were made of the histochemical characteristics of skeletal muscle from 10 animal species. The basic comparison was made from the staining patterns for the myofibrillar actomyosin ATPase produced by preincubation of fresh frozen cross-sections of muscle at alkaline pH (10.30) or acid pH (4.60) with those produced by preincubation in media containing Cu2+ at alkaline pH (10.30), near neutral pH (7.40), or acid pH (4.60). Muscle sections were also stained for reduced nicotinamide adenine dinucleotide tetrazolium reductase and alpha-glycerophosphate dehydrogenase to provide an indicati...
Bottoms GD, Johnson MA, Lamar CH, Fessler JF, Turek JJ.Dispersed equine vascular endothelial cells grown in tissue culture, and freshly isolated neutrophils were used to determine direct effects of endotoxin on cyclooxygenase and lipoxygenase products. Endothelial cells (10(7)/ml) or neutrophils (2 X 10(6)/ml) were incubated with (a) buffer, (b) endotoxin (10 micrograms/ml), (c) endotoxin + flunixin meglumine (10 micrograms/ml), or (d) calcium ionophore, A23187 (10 micrograms/ml). Thromboxane (TxB2), prostacyclin (6-keto-PGF1 alpha), and leukotriene C4 (LTC4) were determined in the incubation fluid by radioimmunoassay. Thromboxane and prostacyclin...
Broström H, Hellström U, Hammarström S, Obel N, Perlmann P.Untreated and neuraminidase-treated equine peripheral blood lymphocytes were analysed for binding of the A hemagglutinin of the snail Helix pomatia (HP). For optimal staining by direct immunofluorescence, the concentration of neuraminidase had to be increased as compared to that needed for other species. Moreover, higher concentrations of HP were required for optimal staining of equine lymphocytes as compared to lymphocytes from other species. Even so, the maximal number of equine lymphocytes exhibiting positive staining was only about 20%. No, or very few, HP-positive lymphocytes were seen wh...
Betton JM, Desmadril M, Mitraki A, Yon JM.The unfolding-refolding transition of horse muscle phosphoglycerate kinase induced by guanidine hydrochloride was studied under equilibrium conditions using four different signals: fluorescence intensity at 336 nm, UV difference absorbance at 286 and 292 nm, ellipticity at 220 nm, and enzyme activity. From the following arguments, we found that the process deviates from a two-state model and intermediates are significantly populated even at equilibrium: (1) the noncoincidence of the transition curves and (2) the asymmetry of the transition curve obtained from CD measurements. From these differ...
Conti A, Godovac-Zimmermann J, Liberatori J, Braunitzer G.beta-Lactoglobulin-like proteins were detected in horse colostrum and normal milk using immunological techniques. In contrast to the beta-lactoglobulins sequenced so far these proteins are monomeric and genetically not homogenous. In this paper we report the first primary structure of a monomeric beta-lactoglobulin from horse colostrum. By means of an automatic liquid-phase sequenator the sequence of peptides obtained by tryptic digestion and by cyanogen bromide cleavage was determined. A limited tryptic digestion and hydrolysis with chymotrypsin provided the necessary overlapping peptides. Th...
Söylemez Z, Ozer I.The inhibition of horse plasma cholinesterase by propranolol showed characteristics which depended upon the identity of the substrate used. With butyrylthiocholine as substrate, the inhibition showed a first-order dependence on inhibitor concentration, and was characterized by a Ki of 8 microM (pH 7.4, 20 degrees C). With p-nitrophenylbutyrate as substrate, a biphasic v-1 versus [I] relationship was obtained. The biphasic curve could be resolved into two components, with apparent Ki's of 9 microM and 1.3 mM. Use of butyrylthiocholine as alternative substrate resulted in partial inhibition of p...
Ellenberger MA, Kaeberle ML, Roth JA.A lymphocyte blastogenic assay was developed to serve as an in vitro correlate of cell-mediated immunity to Rhodococcus (Corynebacterium) equi (R equi) in the equine species. Lymphocytes obtained from a group of experimental ponies showed no response in cell culture to R equi heat extract or lysozyme extract antigens. Ponies were assigned to groups for experimental inoculation. Three ponies were inoculated subcutaneously with live R equi, 3 were given live R equi by intranasal and intratracheal routes, and 4 ponies were left untreated. Lymphocytes from all inoculated ponies had a mitogenic res...
Magnuson NS, Perryman LE, Wyatt CR, Ishizaka T, Mason PH, Namen AE, Banks KL, Magnuson JA.Peripheral blood mononuclear cells (PBMC) from 14 foals with hereditary severe combined immunodeficiency (SCID) were studied to determine the extent of lymphocyte differentiation that occurs in this disorder. PBMC from all 14 horses had the morphologic characteristics of large granular lymphocytes (LGL). Cells from only one of 14 horses were responsive to phytolectin stimulation in a standard blastogenesis assay; however, PBMC from all 14 horses proliferated in continuous culture in the presence of partially purified interleukin 2. Furthermore, there were differences in the growth patterns of ...
Pipers FS, Adams-Brendemuehl CS.To establish the feasibility of utilizing high-frequency ultrasound to image equine fetal development transabdominally, in vitro and in vivo observations were made. Three fetuses retrieved from mares that died from various causes were scanned in vitro to establish fetal images from various orientations. A total of 50 mares ranging in age from 4 to 16 years and representing various breeds were scanned intermittently from 100 days of gestation to parturition. Fetal parts, amniotic fluid, placental membranes, and motion patterns could be reliably identified. Fetal heart rates decreased from 180 b...
Scott DW, Walton DK, Smith CA, Lewis RM.Indirect immunofluorescence testing for pemphigus-like antibodies was performed on 79 horses: 28 horses with various nonpemphigus dermatologic diseases, 21 horses with various nondermatologic diseases, and 30 normal horses. Pemphigus-like antibodies were detected in 6 horses: 3 normal horses with titers of 1:40, 2 horses with dermatophilosis at titers of 1:10 and 1:80, and 1 horse with lymphosarcoma at a titer of 1:320. It was concluded that equine pemphigus-like antibodies are a potential source of misinterpretation and misdiagnosis in indirect immunofluorescence testing. Direct immunofluores...
Coignoul FL, Bertram TA, Cheville NF.Neutrophils isolated from venous blood of adult and foal ponies inoculated with equine herpesvirus-1 were evaluated by in vitro function tests and by electron microscopy. Foals had fever and severe neutropenia 24 hours after inoculation; increased neutrophil random migration under agarose and decreased antibody-dependent cell-mediated cytotoxicity were significant at 24 hours, but values had returned to preinoculation levels by 72 hours. Mares had fever and leukopenia of less severity, increases in neutrophil migration, and longer persistence of primary granule release than were seen in foals....
Papkoff H, Murthy HM, Roser JF.The tyrosine residues of equine chorionic gonadotropin have been nitrated with tetranitromethane and the resulting effects on the biological and immunological activities of the hormone studied. All of the tyrosine residues in equine chorionic gonadotropin were found to react with tetranitromethane when a 100-fold molar excess of reagent was used or with an 8.6 molar excess in the presence of 5 M guanidine hydrochloride. Complete nitration abolished the biological activities and decreased the immunological activity of the hormone. The nitration of one tyrosine residue resulted in the loss of 70...
Blanchard DB, Leid RW.Bovine and equine factor D were purified to apparent homogeneity as evidenced by a single protein staining band on 7.5-17.5% SDS-PAGE slab gels under both reducing and non-reducing conditions. An apparent mol. wt of 15,000 for bovine D and 22,500 for equine D were noted after SDS-PAGE gel analysis of both reduced and non-reduced preparations. A single polypeptide chain for both proteins was evidenced by the lack of any change in the electrophoretic mobility under each of these conditions. The bovine and equine D were enriched 3347- and 9447-fold, with a 20 and 29% yield of hemolytic activity, ...
Shirer HW, Orr JA, Loker JL.To determine if CO2-sensitive airway receptors are important in the control of breathing, CO2 was preferentially loaded into the respiratory airways in conscious ponies. The technique involved adding small amounts of 100% CO2 to either the latter one-third or latter two-thirds of the inspiratory air in an attempt to raise CO2 concentrations in the airway dead space independent of the arterial blood. Arterial blood gas tensions (PCO2 and PO2) and pH, as well as respiratory output (minute volume, tidal volume, and respiratory rate), were measured in a series of 20 experiments on 5 awake ponies. ...
Tzipori S, Withers M, Hayes J, Robins-Browne R, Ward KL.Equine small intestinal brush-border membranes, from 40 adult horses were tested in vitro for the presence of receptors for the Escherichia coli adhesive antigens K88ab, K88ac and K99. Only K88-positive strains of E. coli adhered strongly to horse brush-border membranes. In contrast, a K88-negative mutant strain J2, 2 K99-positive strains and 3 E. coli strains isolated from foals failed to adhere to horse brush-border membranes. Purified K88ac pili when reacted with equine brush-border membranes inhibited to a great extent the adhesion of K88-positive E. coli. Similarly, K88-positive E. coli p...
Makhaeva GF, Shataeva GA, Iankovskaia VL, Fetisov VI, Loshadkin NA.The interaction of human erythrocyte acetylcholinesterase, horse serum butyrylcholinesterase and rat liver carboxylesterase with insecticides (RO)2P(O)SCH(COOEt)SP(O)(OR)2 (I) and (RO)2P(O)SCH(COOEt)OP(S)(OR)2 (II) was studied. The type I and II compounds were not hydrolyzed by carboxylesterase and inhibited the esterases irreversibly. A complex pattern of inhibition of acetylcholinesterase and butyrylcholinesterase by these compounds was caused by kinetically-manifested formation of an enzyme-inhibitor complex. The compounds I and II were more selective towards butyrylcholinesterase than towa...
Province CA, Amann RP, Pickett BW, Squires EL.Two experiments were conducted to evaluate the effects of six extenders and three glycerol levels on the motility of sperm stored at 5 degrees C. Using a split-ejaculated design, semen from 10 dogs and 12 stallions was extended with egg-yolk-tris (EYT), egg-yolk-bicarbonate (EGB), Beltsville F-3 (BF-3), Cornell University (CUE), caprogen (CAP) and heated skim milk (SM) extenders. After cooling to 5 degrees C, additional extender containing 0% to 12% glycerol was added to provide a final concentration of 0%, 3% or 6% glycerol. Regardless of glycerol level, a higher (P<0.05) percentage of can...
Corda M, Pellegrini M, Rinaldi A.Diamine oxidase was prepared from horse kidney by a procedure involving heat denaturation at 50 degrees C, ammonium sulfate fractionation, chromatography on hydroxyapatite and on G-200 Sephadex columns. This procedure gave about 1000 fold purification over the crude kidney cortex homogenate. The enzyme preparations thus obtained are stable only at high ionic strength. The effect on enzyme activity of salt concentration and various stabilizing agents have been investigated. The horse kidney diamine oxidase is irreversibly inhibited by carbonyl reagents and shows substrate specificity quite simi...
Edwards KE, Stevens S, Woodward CB, Tweeten KA.Counterimmunoelectrophoresis was evaluated as a method to distinguish urine of human origin from that of equine origin. The procedure used anti-equine serum and anti-human serum antibodies that had been solid-phase absorbed to eliminate species cross-reactivity. Counterimmunoelectrophoresis reliably detected contamination of equine urine by human urine to a level of 10% with a minimum sensitivity to about 2% contamination. Compared with double diffusion, counterimmunoelectrophoresis was approximately 10 to 15 times more sensitive in the detection of urine proteins.
Robinson-Steiner AM, Beebe SJ, Rannels SR, Corbin JD.Excluding autophosphorylated species, at least six forms of the regulatory subunit of type II cAMP-dependent protein kinase (RII) from various mammalian tissues were identified by sodium dodecyl sulfate (SDS) gel electrophoresis of purified samples and of crude preparations photoaffinity labeled with 8-azido[32P] cAMP and by gel filtration. After autophosphorylation some heart RII forms termed type IIA (bovine, porcine, equine, and dog) shifted to a more slowly migrating band on SDS gels while others termed type IIB (rat, guinea pig, rabbit, and monkey) did not detectably shift. Both subclasse...
Myer YP, Kumar S.The ascorbate reduction of horse heart ferricytochrome c in 0.05 M phosphate + 0.25 M sodium sulfate, at pH 7.3, as a function of temperature, 12-36 degrees C, and at alkaline pH 8.4 using stopped flow technique has been examined. The data have been analyzed in terms of a two-step mechanism, binding followed by reduction (Myer, Y.P., Thallam, K.K., and Pande, A. (1980) J. Biol. Chem. 255, 9666-9673). At neutral pH and up to about 26 degrees C, the first order reduction constant is independent of temperature, i.e. with zero or near-zero activation energy. At higher temperatures, it becomes temp...
Orzoł D, Kępska M, Zyzak M.Equine metabolic syndrome (EMS) is a prevalent endocrine disorder associated with insulin dysregulation, oxidative stress, and impaired regenerative capacity of adipose-derived stem cells (ASCs). The aim of this study was to evaluate the effects of orientin-a plant-derived flavonoid with known antioxidant properties-on equine ASCs (EqASCs) derived from both clinically healthy and diagnosed EMS-affected mares. EqASCs were treated with orientin to evaluate its biological effects. The analysis included key cellular functions such as proliferative capacity, viability, apoptosis, oxidative stress, ...
Hobbs SJ, Mather J, Rolph C, Richards J.Radial strain in normal hooves has been found to vary with strain gauge location, limb posture and sample limb but reported magnitudes were considered to be low. More accurate measurement of radial strain may enhance the understanding of hoof function. Objective: To explore in vitro radial hoof strain in relation other kinetic and kinematic variables that may be related. Methods: Five normal forelimbs were removed at the proximal articular surface of the third metacarpal bone (McIII). The limbs were loaded using a modified Instron test machine. Six calibrated infrared cameras captured movement...
Li GP, Seidel GE, Squires EL.Five experiments were designed to study the fertilizability and development of bovine oocytes fertilized by intracytoplasmic sperm injection (ICSI) with stallion spermatozoa. Experiment 1 determined the time required for pronuclear formation after ICSI. Equine sperm head decondensation began 3 h after ICSI; 42% were decondensed 6 h after ICSI. Male pronuclei (MPN) began to form 12 h after ICSI. Female pronuclei (FPN), however, formed as early as 6 h after ICSI. In Experiment 2, ionomycin, ionomycin plus 6-dimethylaminopurine (DMAP), and thimerosal were used to activate ICSI ova. None of the IC...
Dejardin LM, Arnoczky SP, Cloud GL.During impact, equine hooves undergo viscoelastic deformations which may result in potentially harmful strains. Previous hoof strain studies using strain gauges have been inconclusive due to arbitrary gauge placement. Photoelastic stress analysis (PSA) is a full-field technique which visually displays strains over entire loaded surfaces. This in vitro study identifies normal hoof strain patterns using PSA. Custom-made photoelastic plastic sheets were applied to the hoof surface. The hooves were axially loaded (225 kg) under level and varus/valgus conditions. Strain patterns were video-recorded...
Marcos R, Ribeiro T, Santos M, Correia-Gomes C, Macedo S, Guimarães T, Blielbernicht M, Caniatti M, Rocha A.Uterine lavage (UL) is a routine diagnostic procedure for endometritis. In UL the fluid is centrifuged and the sediment smeared. Samples prepared in cytocentrifuges, the so-called "cytospins", are useful for evaluating cells in fluids, but never been used in UL. The aim of this study was to assess the usefulness of cytospins after UL, comparing automatic versus manual cytocentrifuges, and to determine their value for the diagnosis of endometritis. The study was divided in two parts. Firstly, UL was performed in 16 mares and a small part of the retrieved fluid was cytocentrifuged in an automati...
Roth TL, White KL, Thompson DL, Rahmanian S, Horohov DW.The mechanism by which a horse conceptus-derived immunosuppressive factor (HCS) of M(r) > 100,000 inhibits lymphocyte proliferation was investigated. The factor was obtained from the culture supernatants of 20-day-old horse conceptuses; activity, identified by reduced uptake of [3H]thymidine by mitogen-stimulated lymphocytes, was greatest (P < 0.01) in cultures stimulated by mitogen from pokeweed. HCS also suppressed cell proliferation stimulated by phytohaemagglutinin (P 0.05). Data from a fluorescence-activated cell sorter indicated that supplementation with HCS reduced the number of ...
Sharma AK, Singh TP.Lactoferrin binds two Fe(3+) and two CO(2-)(3) ions with high affinity. It can also bind other metal ions and anions. In order to determine the perturbations in the environments of the binding sites in the N and C lobes and elsewhere in the protein, the crystal structure of oxalate-substituted diferric mare lactoferrin has been determined at 2.7 A resolution. The final model has a crystallographic R factor of 21.3% for all data in the resolution range 17.0-2.7 A. The substitution of an oxalate anion does not perturb the overall structure of the protein, but produces several significant changes...
Menozzi A, Pozzoli C, Poli E, Delvescovo B, Serventi P, Bertini S.We investigated the effects of nonselective muscarinic antagonist (atropine) and of selective muscarinic subtype 1 (M1), 2 (M2), 3 (M3) antagonists (VU0255035, methoctramine, pFHHSiD, respectively) on the contractions evoked by electrical field stimulation (EFS) or by exogenous ACh in isolated horse bronchial muscle. Atropine completely inhibited neurogenic contractions in a concentration-dependent fashion, whereas selective muscarinic antagonists induced relevant modifications only at the highest concentration tested. Experiments with selective muscarinic antagonists in combination showed tha...
Willsallen H, Heller J, Kark L, Hilbert BJ.In vitro comparison of the mechanical properties of braided polyurethane elastomer (Lycra®) and braided polyester (Ethibond™) (1) when inserted into the muscular process of the arytenoid cartilage and (2) as suture loops. Methods: Experimental. Methods: Equine cadaver larynges (n = 15). Methods: The muscular processes (n = 30) of the arytenoid cartilages were dissected from each larynx and embedded in a resin base. Lycra® and Ethibond™ prostheses were randomly allocated to the left or right muscular process and each underwent cyclic fatigue (25-50 N) followed by load-to-failure...
Dettwiler R, Schmitz AL, Plattet P, Zielinski J, Mevissen M.The activity of cytochrome P450 enzymes depends on the enzyme NADPH P450 oxidoreductase (POR). The aim of this study was to investigate the activity of the equine CYP3A94 using a system that allows to regulate the POR protein levels in mammalian cells. CYP3A94 and the equine POR were heterologously expressed in V79 cells. In the system used, the POR protein regulation is based on a destabilizing domain (DD) that transfers its instability to a fused protein. The resulting fusion protein is therefore degraded by the ubiquitin-proteasome system (UPS). Addition of "Shield-1" prevents the DD fusion...
Gonchar MV, Lavrenova GI, Rudenskaia GN, Gaĭda AV, Stepanov VM.Using ion-exchange and affinity chromatography and isoelectrofocusing, eight forms of pepsin with pI 1.6, 1.8, 2.1, 2.3, 2.6, 2.8, 3.2 and 3.6, were isolated from horse gastric juice. The molecular weights, amino acid composition, N-terminal sequence and functional activity of these multiple forms were determined. Partial primary structure of tryptic peptides of pepsin with pI 2.3 was investigated. The analyzed partial sequences of the forms with pI 1.8, 2.1, 2.3, and 2.6 have identical structures which differ from the amino acid sequence of pepsin with pI 3.2 by four substituents. In terms of...
Katano Y, Imai S.The effects of dipyridamole on thromboxane A2 formation by horse platelet microsomes were studied in comparison with those of imidazole, a prototype inhibitor of TXA2 synthetase and nifedipine, a calcium antagonistic vasodilator. Thromboxane A2 was synthesized by incubating PGH2 with horse platelet microsomes and was assayed on the superfused rabbit aorta. Dipyridamole induced as strong an inhibition of TXA2 synthesis as imidazole, while nifedipine was without effects. The possible beneficial clinical outcomes of this effect of dipyridamole are discussed.
Bailey SR, Andrews MJ, Elliott J, Cunningham F.Platelets are thought to play a role in equine diseases such as acute laminitis and verminous arteritis and may be involved in allergic disease. Mediators implicated in the pathophysiology of these conditions activate platelets and responses may be enhanced by interactions between mediators. The present study compared platelet aggregation, thromboxane production and release of radiolabelled [(3)H]5- HT in response to 5- HT, histamine, ADP and PAF alone and in combination in vitro.PAF caused concentration-related aggregation, [(3)H]5- HT release and thromboxane production. In contrast, ADP caus...
Casella S, Giannetto C, Giudice E, Piccione G.The objective of the present study was to evaluate the platelet response to different workload in athletic horse and the influence of hydrocortisone on this response. For this purpose, six Sella Italiana gelding athletic horses, ranging in age from 8 to 10 years and clinically healthy, were subjected to four different exercises: horse-walker, treadmill, obstacle course of 1.00 m and obstacle course of 1.25 m. From all the horses, citrated plasma samples were collected at rest, immediately after exercise and 30 min after the end of the exercise to assess the maximum degree of platelet aggregati...
Aguiar CS, Barros CHSC, Machado WM, Allaman IB, Leite AO, Barbosa LP, Snoeck PPDN.The aim of this study was to evaluate the association of different concentrations of Trolox and the addition of a fixed concentration of DHA in the freezing of semen of Mangalarga Marchador stallions. To that end, 16 ejaculates were frozen in the following extenders: E1) BotuCrio (BC; Control); E2) BC + 50 ngml DHA + 30 µM Trolox (BCDHA30T); E3) BC + 50 ngml DHA + 40 µM Trolox (BCDHA40T); E4) BC + 50 ngml DHA + 50 µM Trolox (BCDHA50T). All the tested extenders were similar in preserving different kinematic parameters, cell functional integrity, compacted DNA, and high and intermediate mitoc...
Gysens L, Depuydt E, Patruno M, Haspeslagh M, Spaas JH, Martens A.Sarcoids are the most common equine skin tumours Although they do not metastasize, they can be locally aggressive and cause significant clinical symptoms in affected horses. Despite being common, very little is known about the host immune response and the biological mechanisms underlying persistence and recurrence of equine sarcoids. The latter reflects the need for further research in this field. This in-vitro study used sarcoid explants from horses with naturally occurring sarcoids (n = 12) to evaluate the induction of a humoral immune response directed against equine sarcoid-derived bovin...
Van Hoogmoed LM, Snyder JR, Nieto J, Harmon FA.To determine whether a customized solution could attenuate the effects of low-flow ischemia and reperfusion injury of the equine jejunum. Methods: A segment of jejunum obtained from 21 healthy adult horses. Methods: A segment of jejunum was maintained in an isolated extracorporeal circuit, and arterial flow was reduced to 20% of baseline for 40 minutes (ischemia) followed by 60 minutes of reperfusion. In 1 group, a customized solution was infused at a rate of 1 ml/min during low-flow ischemia and 3 ml/min during reperfusion. In a second group, the solution was infused at the same rate during l...
Fatemi-Nainie S, Anderson LW, Cheevers WP.MC-1 is an equine sarcoid-derived cell line which spontaneously releases a retrovirus possessing genomic sequence homology with an inducible endogenous retrovirus of normal equine cells. A complete characterization of MC-1 tumor cells was undertaken, including morphology, growth kinetics, and saturation density, selective growth in semisolid media, uptake of 2-deoxyglucose, and tumorigenicity in athymic nude mice. MC-1 cells, in contrast to normal equine dermal fibroblasts, exhibit all of the characteristics of malignantly transformed cells.
Katz LM, Marr CM, Elliott J.To compare responses of equine digital arteries (EDAs) and veins (EDVs) to human-αcalcitonin gene-related peptide (hαCGRP), evaluate effect of the endothelium, and characterize receptors and sources of endogenous CGRP. Methods: Palmar digital vessels (5 to 9/experiment) from healthy adult horses killed at an abattoir. Methods: Vessel rings were mounted under tension in organ baths containing Krebs-Henseleit solution at 30 °C, with relaxation responses examined in vessels preconstricted with a thromboxane-mimetic (3 × 10(-8)M). Responses of endothelium-intact (+e) and -denuded (-e) EDAs and...
Malek G, Richard H, Beauchamp G, Laverty S.Focal bone microcracks with osteoclast recruitment and bone lysis, may reduce fracture resistance in racehorses. As current imaging does not detect all horses at risk for fracture, the discovery of novel serum biomarkers of bone resorption or osteoclast activity could potentially address this unmet clinical need. The biology of equine osteoclasts on their natural substrate, equine bone, has never been studied in vitro and may permit identification of specific biomarkers of their activity. Objective: (1) Establish osteoclast cultures on equine bone, (2) Measure biomarkers (tartrate resistant ac...
Gray PR, Derksen FJ, Robinson NE, Slocombe RF, Peters-Golden ML.We have developed an alternative method for examining equine tracheal epithelial arachidonic acid (AA) metabolism that utilizes strips of pseudostratified columnar epithelium attached to a layer of elastic tissue 80 to 130 microns thick. We compared the responses of this preparation with those of enzymatically dispersed suspensions of tracheal epithelium obtained from the same animal. Strips incubated with [3H]AA incorporated 40.8 +/- 3.6% of added radioactivity and released 2.55 +/- 0.23% of incorporated radioactivity when stimulated with 5 microM A23187. Values for the cell suspension were 5...
Weaver MP, Shaw DJ, Munaiwa G, Fitzpatrick DP, Bellenger CR.The objectives of this study were to topographically map pressure distribution across the palmar surface of the navicular bone in response to forces applied by the deep digital flexor tendon (DDFT), and evaluate the effect of raising the heels in vitro . A rig was constructed to hold an equine cadaver limb and apply controlled pressure to the navicular bone from the DDFT. Pressure-sensitive paper was used to quantify and map the resultant pressure. Changes in response to heel wedges (5 degrees and 10 degrees ) were recorded. The areas subject to maximum pressure from the DDFT were: the central...
Poole AW, Heath MF, Sage SO, Evans RJ.Equine platelets in platelet-rich plasma were incubated with the fluorescent indicator dye, Fura-2-AM (Fura-2-acetoxymethyl ester) and the degree of loading of the cells with the dye and the extent of hydrolysis of the ester was assessed by quantitative fluorimetry and by thin-layer chromatography respectively. Under these conditions the cells loaded poorly with Fura-2 to a concentration of 4 microM. The technique was validated by demonstrating adequate loading of human platelets with Fura-2, to a concentration of 250-300 microM, using the same method. The removal of plasma from the extracellu...
Joppich-Kuhn R, Luisi PL.The interaction between horse liver alcohol dehydrogenase and the oxidized and reduced forms of the 3-thionicotinamide--adenine dinucleotide coenzyme analogues (sNAD and sNADH) has been investigated by ultraviolet absorption, fluorescence and circular dichroism. The fluorescence of sNADH is enhanced when bound to the enzyme, and the protein fluorescence is quenched by both sNADH (60--65%) and sNAD (65%). The possible origin of the larger quenching produced by sNAD with respect to that of NAD is discussed. Coenzyme dissociation constants have been determined by monitoring the quenching of the p...
Delbeke FT, Teale P, Debackere M, Houghton E.A commercially available generic promazine ELISA kit is available which shows cross-reactivity for the tranquilizer chlorprothixene (CPT). The ELISA test readily detects the presence of CPT or its metabolites in equine urine for up to 24 h after the i.v. and i.m. administration of sub-therapeutic doses (4.5 mg) to three horses. Maximum concentrations (CPT equivalents) are obtained 2 h after i.v. dosing. No distinct concentration peak values are observed after i.m. administration. Following solid-phase extraction, confirmation of CPT and its metabolites by electron impact mass spectrometry afte...
Tappenbeck K, Schmidt S, Feige K, Naim HY, Huber K.Lidocaine is the most commonly chosen prokinetic for treating postoperative ileus in horses, a motility disorder associated with ischaemia-reperfusion injury of intestinal tissues. Despite the frequent use of lidocaine, the mechanism underlying its prokinetic effects is still unclear. Previous studies suggested that lidocaine altered cell membrane characteristics of smooth muscle cells. Therefore, the present study aimed to elucidate effects of lidocaine administration on characteristics of detergent-resistant membranes in equine jejunal smooth muscle. Lidocaine administration caused significa...
Gulevskiĭ AK, Grishchenko VI, Tereshchenko OS, Zagnoĭko VI, Akhremenko AK, Kucherenko SN.The effect of the fraction (1-10 kDa) obtained from the brain of cold-adapted animal (Yakut horse) on Ca2+ transport in sarcolemma vesicles of cardiomyocytes was investigated. It was shown that during insertion of Yakut horse brain fraction into incubation medium at the concentration from 10(-9) M to 3.10(-5) M at Ca2+ transport substrate concentration from 0.1 mM to 1.0 mM, the rate of Ca2+ passive penetration into vesicles slightly increased and at Ca2+ transport substrate concentration 3 mM, which is physiologic, a decrease of rate values was established for all concentrations of the fracti...
Mallem MY, Thuleau A, Noireaud J, Desfontis JC, Gogny M.To investigate the role of superoxide anions in the lipopolysaccharide (LPS)-induced impairment of beta-adrenoceptor-mediated equine digital vein (EDV) vasodilation. Methods: EDVs isolated from forelimbs of 24 healthy adult horses. Methods: Endothelium-intact or endothelium-denuded EDV rings were incubated with or without LPS (10 microg/mL) of Escherichia coli (O55:B5) for 4 hours. Cumulative concentration-relaxation curves resulting from administration of isoprenaline, a nonselective beta-adrenoceptor agonist, or from administration of SR 58611A, a selective beta(3)-adrenoceptor agonist, were...
Klein C, Donahue JM, Sells SF, Squires EL, Timoney PJ, Troedsson MH.To determine the impact of antimicrobial-containing semen extender on the growth of Taylorella equigenitalis in semen culture-positive for contagious equine metritis (CEM) and the development of CEM after artificial insemination with CEM-positive semen extended with antimicrobial-containing semen extender. Methods: Prospective study. Methods: 21 mature mares free of CEM, 1 mature stallion experimentally infected with CEM, and semen from a stallion naturally infected with CEM. Methods: CEM-positive semen was incubated in semen extender with and without antimicrobials (amikacin [final concentrat...
Skok MV, Denisiuk PV, Komissarenko SV.Glutaraldehyde treatment does not change the absorption of cytochrome c either in the visible or in UV spectra. It brings about the formation of dimers, trimers and high-polymeric forms of cytochrome c and shifts the pI of all cytochrome c isoelectric fractions to more acid pH. Polymerization also results in changes of kinetic parameters of cytochrome c benzidine reaction increasing its affinity to 3,3-diaminobenzidine with a simultaneous decrease in the effectiveness of H2O2 binding. These biochemical changes can be related to immunochemical differences of native and glutaraldehyde-treated cy...
