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.
Allen GP, Yeargan MR, Bryans JT.The effect of in vitro and in vivo serial virus passage on the genetic stability of equine herpesvirus 1 (EHV-1) was investigated by restriction endonuclease analysis of the viral DNA. DNAs of EHV-1 isolates at different passage levels in cultured cells or in Syrian hamsters were compared by electrophoresis of the DNA cleavage fragments produced by restriction endonuclease digestion. No changes were observed in the restriction profile of the DNAs of EHV-1 strains after 100 sequential passages in cultured equine cells. However, serial passage of the virus in hamsters or in cells of non-equine o...
Ginther OJ.Fixation and orientation of the conceptus within the uterine lumen of 40 barren mares were examined by ultrasound daily on days 11-21 and at three-day intervals thereafter until day 48. The growth curve of the in situ conceptus had a distinct plateau between days 17-24, as determined by measurements of the width and area of conceptuses on the ultrasound images. The vesicle expanded at an average daily rate of 3-4 mm before the plateau and 2-3 mm after the plateau. Dramatic changes occurred in the shape of conceptus. The predominant shapes were approximately as follows: days 11-16, spherical; d...
Nara PL, Krakowka S, Powers TE, Garg RC.Fourteen young outbred horses, divided into 2 groups on the basis of 18- or 24-hour skin-test reactions to Streptococcus equi, were inoculated nasopharyngeally with virulent S equi. Animals (n = 6, group I) with evidence of previous exposure to S equi (positive dermal response and existing serum antibodies), with one exception, developed minimal or no signs of disease after inoculation. In contrast, S equi skin-test negative and seronegative horses (n = 8, group II) developed predictable and severe clinical signs of infection after their inoculation, including shedding of the organism from nas...
Ralston JS, Main AR, Kilpatrick BF, Chasson AL.Two large-scale methods based primarily on the use of procainamide-Sepharose gels were developed for the purification of horse and human serum non-specific cholinesterases. With method I, the procainamide-Sepharose 4B gel was used in the first step to handle large volumes of serum. With method II, the procainamide-Sepharose 4B gel was used in the final step to obtain pure enzyme. Although both methods gave electrophoretically pure cholinesterase preparations in good yields, they were significantly more efficient at purifying the horse enzyme than the human enzyme. To study this problem, the re...
Zweygarth E, Ahmed JS, Rehbein G, Voigt WP.The capacity of equine peripheral blood lymphocytes (PBL) to proliferate in the presence of Babesia equi-transformed lymphoblastoid stimulator cells was tested in an autologous as well as in an allogenic one way mixed lymphocyte reaction (MLR). It was found that both autologous and allogeneic responder lymphocytes incorporated high amounts of 3H-thymidine. The incorporation of 3H-thymidine was lower in MLR using as stimulator cells lymphocytes from which the cell line had previously been established, than when using parasitized culture cells as stimulator. Proliferation of PBL was achieved onl...
Johnstone IB.Nonsteroidal anti-inflammatory drugs are commonly used in the treatment of inflammatory conditions, and have potential value in the treatment of thrombotic disease in the horse. This study compares the potency of three nonsteroidal anti-inflammatory drugs phenylbutazone, naproxen (equiproxen) and flunixin meglumine (banamine) with respect to their effects on equine platelets. Two functional responses of horse platelets were evaluated in vitro: their ability to aggregate and their ability to make available platelet factor 3 procoagulant activity. Flunixin at a concentration of 10(-6) M signific...
Leblová S, Galociová J, Cerovská N.The studied herbicides (terbutylazine, simazine) inhibit the activity of plant, animal, and yeast alcohol dehydrogenases. The inhibition constant Ki for alcohol dehydrogenase (ADH) isolated from peas and bakers' yeast equals approximately 10(-4) M, and that for ADH isolated from horse liver is of the order of 10(-5) M. The character of inhibition for all the herbicides studied for the reaction catalyzed by pea, liver, and yeast ADH is always noncompetitive toward ethanol and competitive with respect to NAD. The inhibition constants for the enzyme isolated from peas are pH independent. The inte...
Igwe OJ, Blake JW.The disposition kinetics of pemoline after iv and oral administration of 2.4 mg/kg of the drug were studied. The elimination half-life was 39.4 hr. The mean volume of distribution was 1.5 liters/kg indicating extensive tissue distribution and sequestration for an amphoteric drug. Plasma protein binding determined by in vitro equilibrium dialysis was concentration dependent. The mean binding capacity was found to be 0.80 mu-mol/g, an apparent dissociation constant of 3.73 X 10(-5) molar, and a total plasma protein concentration of 64.7 g/liter. The mean systemic availability by oral administrat...
Brimijoin S, Mintz KP, Prendergast FG.Interactions between dansylarginine N-(3-ethyl-1,5-pentanediyl)amide (DAPA) and the cholinesterases were examined by the techniques of enzyme kinetics and fluorescence spectroscopy. When tested with partially purified enzyme preparations, DAPA was a potent inhibitor of butyrylcholinesterase (IC50 = 2 x 10(-7) M) but not of acetylcholinesterase (IC50 = 4 x 10(-4) M). For a detailed study of the effects of DAPA on butyrylcholinesterase (BuChE), the enzyme was purified to homogeneity from horse serum, with the aid of affinity chromatography on N-methyl acridinium. The kinetics of the inhibition o...
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.
Chin WW, Maizel JV, Habener JF.The sizes of the human and subhuman alpha-subunits of the glycoprotein hormones differ by four amino acids (hCG alpha, 92 amino acids; murine, equine, bovine, and ovine alpha, 96 amino acids). The shortening of the human alpha-subunit has been attributed to posttranslational proteolysis. We have recently determined the nucleotide sequences of the mRNAs encoding the precursors of the alpha-subunit of mouse TSH and rat gonadotropins using recombinant DNA techniques. In this report, we have compared these nucleotide sequences and their deduced amino acid sequences with those of the pre- alpha-sub...
Billah MM, Lapetina EG.Stimulation of horse platelets with platelet-activating factor (PAF) induces a rapid degradation of phosphatidylinositol 4,5-bisphosphate [PtdIns(4,5)P2]. Addition of 0.1 microM PAF for 5 sec to platelets prelabeled with 32P induces a 50% loss of [32P]PtdIns(4,5)P2. 32P-Labeled phosphatidylinositol 4-monophosphate (PtdIns4P) and [32P]phosphatidylinositol (PtdIns) also are decreased, albeit at a slower rate. Loss of 32P radioactivity correlates with a net loss of fatty acids from both polyphosphoinositides. Stimulation of platelets with PAF also produces formation of [32P]phosphatidic acid and ...
Camp CJ, Leid HW.Extracts of Onchocerca cervicalis, an equine parasite, were incubated with radiolabeled equine neutrophils and neutrophil migration was assessed for factors derived from the parasite itself or for host-derived factors after incubation of these same parasite extracts with equine serum. No stimulus for cell migration was observed in saline extracts of adult worms, uterine microfilariae, or skin microfilariae at any dosage tested. However, after incubation of saline extracts with fresh normal equine sera a marked stimulus for neutrophil migration was observed. Ablation of this biologic activity w...
McLaughlin SA, Brightman AH, Helper LC, Manning JP, Tomes JE.In 123 cases of external ocular disease in the horse, pathogenic microorganisms isolated with greatest frequency were Streptococcus sp (43.9%), Staphylococcus sp (24.2%), and Pseudomonas sp (13.8%). Fungi were isolated in 4.8% of the cases. In vitro testing showed that most of the Streptococcus sp isolants were sensitive to ampicillin, cephalothin, and carbenicillin. Most of the Staphylococcus sp isolants were sensitive to gentamicin, cephalothin, and bacitracin. Most of the Pseudomonas sp isolants were sensitive to polymyxin B and gentamicin.
Moltmann UG, Mehlhorn H, Schein E, Rehbein G, Voigt WP, Zweygarth E.The development of Babesia equi (Piroplasmia) in the vertebrate host was studied by electron microscopy. The tick-borne sporozoites initiated an exoerythrocytic schizogony in lymphocytes in vivo and in vitro. In lymphocyte cultures the schizonts of B. equi lay as fissured bodies directly within the host cell forming numerous cytomeres. These cytomeres were bordered by a single membrane and contained polymorphous nuclei. Merozoite differentiation was initiated at several places by protrusions appearing at the periphery of the cytomeres. The cytoplasm of the schizont merged progressively into th...
Montelaro RC, West M, Ivey M.Representative glycoproteins including fetuin, protein A, ovalbumin, alpha 1 acid glycoprotein, and the major glycoprotein of equine infectious anemia virus were labelled with 125I by the chloramine-T or Bolton-Hunter procedure and their binding to immobilized Con A or lentil lectin compared to untreated samples of each glycoprotein. Glycoprotein modification was no greater than one substituted residue per protein molecule. Yet the radioiodinated glycoproteins typically displayed only 0-50% of the lectin binding observed with untreated samples. These results indicate that lectin glycoprotein b...
Pellegrini A, Zweifel HR, von Fellenberg R.The alpha-2-protease inhibitor, of 65,000 daltons molecular weight, described by several authors in horse plasma and also present as a contaminant in alpha-1-isoinhibitor isolates previously described by us (Pellegrini & von Fellenberg (1980) Biochim. biophys. Acta 616, 351-361) has now been isolated to purity and identified as antithrombin III. The inhibitor is composed of a single polypeptide chain as judged by SDS polyacrylamide gel electrophoresis. The inhibitor was effective only against trypsin and thrombin. Serological cross-reaction existed between the inhibitor and the antiserum t...
Dutta SK, Myrup AC.A novel, simple method of infectious center assay was developed to detect and quantitate the intracellular existence of equine herpesvirus 1 and equine herpesvirus 2 in peripheral blood mononuclear cells infected in vivo and in vitro with the viruses by cocultivation of these cells with a permissive equine cell culture. The infectious center titers were correlated with the infectious virus titers. In vivo equine herpesvirus 1-infected mononuclear cells obtained from ponies experimentally infected with the virus and equine herpesvirus 2-infected mononuclear cells obtained from selected naturall...
Pellegrini A, Zweifel HR, von Fellenberg R.Several publications have described in the past properties of partly purified horse alpha 2-macroglobulin (alpha 2M) which are strikingly different from the human alpha 2M. Horse alpha 2M was therefore isolated to purity by classical procedures, i.e. affinity chromatography, ion exchange chromatography and gel filtration, and its properties are compared with those of its human counterpart. The molecular weight of the native protein and its subunits, the isoelectrofocusing pattern and the change in electrophoretic mobility caused by interaction with protease were similar to those of human alpha...
Latif ZA, Fletcher MA.Immunochemical comparisons were made of the reactivity of membrane glycoproteins from horse, bovine, sheep and goat erythrocytes with heterophile antibodies of infectious mononucleosis. The four receptors were tested as competitive inhibitors of a sandwich-type solid-phase radioimmunoassay and of agglutination of glycoprotein-latex reagents by infectious mononucleosis serum. The results of this study showed that the bovine glycoprotein had a superior reactivity with this heterophile antibody system and sheep erythrocyte glycoprotein was the least reactive. The latter had negligible ability to ...
Peters GJ, Oosterhof A, Veerkamp JH.1. Activity of uridine kinase was very low in ovine lymphocytes and in those of some pigs. Lymphocytes of other pigs showed a significantly higher activity of this enzyme. Activity of uridine kinase in lymphocytes of man, horse and cattle was intermediate. 2. Activity of uridine phosphorylase was higher than that of uridine kinase with lymphocytes of all species. 3. Activity of uridine kinase in equine lymphocytes increases at PHA-stimulation and also in porcine lymphocytes with a low activity at the start of the culture. Activity of uridine kinase decreased in porcine lymphocytes with a high ...
Marcum JA, Highsmith RF, Kline DL.Complete activation of purified horse plasminogen to plasmin was obtained with a 1:10 molar ratio of streptokinase to plasminogen after 5 min of incubation at 37 degrees C. At a 1:1 molar ratio, maximal activity did not appear until 15-30 min, while at a ratio of 6:1 complete activation was delayed for 120-180 min. Gel filtration studies of isotopically labeled streptokinase and horse plasminogen suggest that the delay was due to impaired formation of a streptokinase-plasminogen complex. The predominant streptokinase moiety within the streptokinase-plasmin complex which forms from the streptok...
Campbell TM, Studdert MJ.An inactivated equine adenovirus type 1 (EAdV1) vaccine was administered to 4 horses. The horses had virus-neutralizing (VN) antibody titers before they were vaccinated, but developed higher VN antibody titers in response to vaccination. Nonvaccinated control horses did not show increases in VN antibody during the study, indicating that any increase in antibody titer in vaccinated horses was a result of vaccination and not due to an EAdV1 epizootic during the study. Specific EAdV1 in vitro lymphocyte blastogenesis (LB) was evaluated, using lymphocytes from 4 vaccinated and 2 control horses. Ho...
Jacobsen K, Gintz T, Reed SM, Newbry J, Bayly WM, Perryman LE, Leid RW.Equine neutrophils (PMN) were isolated to greater than 99% purity by isopyknic sedimentation on coated colloidal silica particles. A cell recovery of 84.7 +/- 4.0%, with a viability of greater than 99%, was observed with this method. The isolated PMN were compared with mixed population of equine peripheral leukocytes with respect to functional integrity by chemiluminescence and bactericidal assays. There was no significant difference (P less than 0.01) observed in either assay between the isolated equine PMN and the mixed-cell populations. The methods used in both the isolation as well as the ...
Read NG, Trist DG.The uptake of mepacrine by isolated horse polymorphonuclear leucocytes (PMN) was measured using spectrophotofluorimetry. Two phases of uptake were observed, the first, rapid fraction, essentially complete by 10 min, and a second, slow fraction, which was still proceeding after 60 min. The appearance of mepacrine within the PMN was also visualized by fluorescence microscopy. Discrete yellow points of fluorescence were observed in the cytoplasm of PMN within 30 s. These discrete points corresponded both in size and number to the PMN granules. After 5 min, the nuclei showed faint fluorescence whi...
Bertram TA, Coignoul FL, Jensen AE.Equine neutrophils were combined with Haemophilus equigenitalis (contagious equine metritis organism; CEMO) or Escherichia coli in low- and high-antibody-titer serum to evaluate the neutrophils ability to phagocytize and kill these bacteria. More E. coli than CEMO were phagocytized at each time period. After 120 min in low-antibody-titer serum, 56.3% of the E. coli and 34.3% of the CEMO were phagocytized. A total of 45% of CEMO and 74.9% of E. coli were phagocytized by 120 min when neutrophils were in high-antibody-titer serum. More than 75% of the ingested E. coli and 90% of the ingested CEMO...
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...
Bertram TA, Coignoul FL.Equine neutrophils were evaluated ultrastructurally and by morphometric analysis. Homogeneous populations of neutrophils were isolated from peripheral blood at 4 degrees and 22 degrees C by centrifugation on two sequential Ficoll-Hypaque density gradients. Isolation procedures at both temperatures resulted in neutrophil degranulation but not cell swelling. Degranulation was more extensive in cells isolated at 22 degrees C. Isolation temperature affected the neutrophil content of secondary granules more than primary granules. A granule similar to immature specific granules of human neutrophils ...
Blikslager AT, Tate LP, Weinstock D.Effects of neodymium:yttrium aluminum garnet (Nd:YAG) laser irradiation on equine endometrium were evaluated in vitro and in six mares with endometrial cysts. The Nd:YAG laser was applied to six endometrial sites, in each of five uterine specimens, with power densities of 5659 to 33,954 J/cm2. Depth of tissue ablation was measured and graded on histologic sections of the tissue lesions. Power density had a significant effect on the depth of tissue ablation (p < .001). Grade 3 lesions (full-thickness ablation of the endometrium) were created with energy densities of 16,977 to 33,954 J/cm2. Six ...
Van Hoogmoed L, Rakestraw PC, Snyder JR, Harmon FA.To determine the in vitro effect of various prostaglandins (PG) and nonsteroidal anti-inflammatory drugs (NSAID) on contractile activity of the large-colon taenia of horses. Methods: 14 healthy horses. Methods: The taenia was collected from the ventral colon, cut into strips (2 X 10 mm), and mounted in a tissue bath system (20-ml capacity) that contained oxygenated Krebs buffer solution warmed to 37.5+/-0.5 C. After equilibration, incremental doses of PGE2, PGF2alpha, PGl2, flunixin meglumine, carprofen, ketoprofen, and phenylbutazone were added to the baths, and contractile activity was recor...
Jarvis GE, Evans RJ.Endotoxaemia is a leading cause of death among horses. Thrombocytopenia is a common finding in clinical and experimentally-induced cases of endotoxaemia and can lead to coagulopathies, including disseminated intravascular coagulopathy which is usually fatal. In this study it was shown that endotoxin (3 ng ml-1 to 25 micrograms ml-1) can aggregate equine platelets in heparinised whole blood in vitro. The endotoxin-induced aggregation (EIA) was shown to be dependent on the presence of leucocytes in the blood and did not occur when detoxified endotoxin was used, suggesting that lipid A was necess...
van Weeren PR, van den Bogert AJ, Barneveld A, Hartman W, Kersjes AW.The function of the reciprocal apparatus in the hind limb of the horse was studied by kinematic gait analysis. For recording purposes a modified opto-electronic CODA-3 kinematic analysis system was used. The raw kinematic data were corrected for skin displacement artifacts by use of recently developed correction models. It was concluded that contradictory findings about the coupling of tarsal and stifle joints by the reciprocal apparatus, when comparing in vitro and in vivo studies, can be fully attributed to artifacts due to the movement of the skin markers over the underlying bony structures...
Watson TD, Burns L, Packard CJ, Shepherd J.Affinity chromatography on heparin sepharose was used to identify 2 lipolytic enzymes in heparinized plasma from horses. One enzyme was typical of hepatic triglyceride lipase (HTGL), because it was resistant to inactivation by high concentrations of NaCl, and it did not require the addition of serum for activity. The other enzyme was identified as lipoprotein lipase (LPL), because of its inactivation at NaCl concentrations in excess of 0.2M, and its dependency on addition of serum as a source of apolipoprotein C-II activator. The enzymes were purified by 347-(HTGL) and 442- (LPL) fold, with yi...
Hampson BA, de Laat MA, Mills PC, Pollitt CC.To determine the effect of various environmental conditions on the degree of hydration in hoof wall horn tissue from feral horses and investigate the effect of short-term foot soaking on moisture content in hoof wall and sole tissue in domestic horses. Methods: 40 feral horses from 3 environments (wet and boggy [n = 10], partially flooded [20], and constantly dry desert [10]) and 6 nonferal Quarter Horses. Methods: The percentage of moisture content of hoof wall samples from feral horses was measured in vitro. In a separate evaluation, the percentage of moisture content of hoof wall and sole t...
Henderson KM, Eayrs K.To develop a means of determining pregnancy status in horses based on measuring serum oestrone sulphate (OS) concentrations using a rapid lateral flow immunoassay, and to determine the assay's effectiveness using a visual end-point. Methods: Serum samples from mares >100 days post-mating (n=701) were assayed using a nitrocellulose membrane-based lateral flow immunoassay device. The device was developed using membrane-bound 1,3,5 (10)-estratrien-3-ol-17-one conjugated to bovine serum albumin as the capture antigen, and an OS-detection monoclonal antibody coupled to colloidal gold as the visi...
Lee OJ, Koch TG.Inflammation-associated disorders are significant causes of morbidity in horses. Equine single-donor mesenchymal stromal cells (sdMSCs) hold promise as cell-therapy candidates due to their secretory nonprogenitor functions. This has been demonstrated by mononuclear cell suppression assays (MSAs) showing that sdMSCs are blood mononuclear cell (BMC) suppressive in vitro. sdMSCs derived from umbilical cord blood are of clinical interest due to their ease of procurement, multipotency, and immunomodulatory ability. Due to the inherent donor-to-donor heterogeneity of MSCs, the development of robust ...
Stewart F, Leigh SE, Thomson JA.Complementary (c)DNA libraries have been made from horse pituitary gland and endometrial cup tissues with the aim of isolating the genes for the horse gonadotrophins (FSH, LH and CG) and growth hormone (GH). Southern (DNA) and Northern (RNA) blotting techniques were used to demonstrate that several heterologous (human and ovine) cDNA probes would be adequate for isolating the horse genes. A human cDNA probe was then used to isolate the horse gonadotrophin alpha-subunit cDNA from the pituitary and endometrial cup libraries. The nucleotide sequences from both tissue sources were identical, there...
Thway TM, Wolfe MW.Primates and equids are the only species known to express the placental glycoprotein hormone, chorionic gonadotropin (CG), a heterodimeric glycoprotein composed of an alpha subunit linked to a hormone-specific beta subunit. The regulatory mechanisms involved in the induction of equine glycoprotein alpha subunit gene expression have not been identified. Epidermal growth factor (EGF) receptor is known to transduce signals that alter a number of different cellular functions (cell proliferation, differentiation, hormone secretion, and gene regulation). In the present study, we investigated the reg...
Martínez AC, Rivera L, Raposo R, García-Sacristán A, Benedito S.The effect of histamine (10(-9)-10(-3) M) on horse penile dorsal artery was evaluated. Precontracted vessels showed a biphasic response (relaxation-contraction) to histamine, while at basal tone, histamine only induced a contractile effect. The H1 receptor agonist, 2-pyridylethylamine (PEA) (10(-9)-10(-3) M), induced concentration-dependent relaxation in precontracted rings and provoked vasoconstriction at basal tone. Mepyramine (10(-9)-10(8) M), an H1 receptor antagonist, competitively antagonized the relaxant response to histamine (pA2 = 9.7) and PEA (pA2 = 9.2). At basal tone, mepyramine (1...
Paik WK, Farooqui J, Gupta A, Smith HT, Millett F.The present observations are the continuation of our earlier study on the physicochemical mechanism of protein-lysine methylation. In this paper the electrophoretic behaviour (pI values) of two chemically modified horse heart cytochromes c at lysine-72 with trifluoromethylphenylcarbamoyl (neutral group) or carboxydinitrophenyl (acidic group) is compared with the enzymatically methylated cytochrome c. The results indicate that although both chemically modified cytochromes c have lower pI values than the unmodified cytochrome c, the enzymatic methylation appears to be much more efficient in lowe...
Whitcomb RW, Schneyer AL, Roser JF, Hughes JP.Using a LH radioligand receptor assay (RRA) previously validated for use in serum and an equine monoclonal RIA, we have distinguished a subset of subfertile stallions with an elevated RRA/RIA ratio. After purification of the active moiety by anion exchange chromatography and immunoprecipitation with the equine LH (eLH) monoclonal antibody, RRA activity remained in the supernatant. This activity was also recognized by a polyclonal LH antibody (GDN 15) with wide cross-species recognition. This active fraction was further purified by gel filtration chromatography and shown to displace labeled eLH...
Auer DE, Ng JC, Seawright AA.Free radical oxidation products, namely conjugated dienes, ultraviolet fluorescence (excitation 325 nm, emission 395 nm) and visible fluorescence (excitation 360 nm, emission 460 nm) were measured in equine synovial fluid exposed to free radicals in vitro and in the plasma and synovial fluids of horses with synovial effusions. The synovial effusions were induced by intra-articularly administered carrageenin (0.3 ml, 1%), which rarely resulted in clinical lameness. The free radicals were generated in vitro by mixtures of iron and ethylene diamine tetra acetate (Fe/EDTA) or mixtures of hypoxanth...
Scarlet D, Budik S, Aurich C.A new device for storage and shipping of cell cultures--the Petaka G3 cell management device--was tested for its applicability for cooled-storage of equine semen. Semen from three stallions was processed with EquiPro extender either without antibiotics (three ejaculates per stallion) or with gentamicin (250 mg/l; three ejaculates per stallion). Semen was either stored at five (anaerobic conditions) or 15 °C (aerobic conditions) in syringes or cell culture devices. Total and progressive motility, as well as membrane integrity of spermatozoa, were evaluated from days 1 to 7 after collection wit...
Sod GA, Mitchell CF, Hubert JD, Martin GS, Gill MS.To compare monotonic biomechanical properties and fatigue life of a 5.5 mm broad limited-contact dynamic compression plate (5.5-LC-DCP) fixation with a 4.5 mm broad LC-DCP (4.5-LC-DCP) fixation to repair osteotomized equine third metacarpal (MC3) bones. Methods: In vitro biomechanical testing of paired cadaveric equine MC3 with a mid-diaphyseal osteotomy, stabilized by 1 of 2 methods for fracture fixation. Methods: Adult equine cadaveric MC3 bones (n=18 pair). Methods: MC3 were divided into 3 test groups (6 pairs each) for: (1) 4-point bending single cycle to failure testing; (2) 4-point bendi...
Hänni K, Hesford F, Lazary S, Gerber H.Genomic DNA isolated from 20 horses was digested with up to six restriction endonucleases and subjected to southern blot hybridization analysis using various human class II alpha- and beta-chain cDNA probes. A high degree of restriction fragment length polymorphism (RFLP) was found for the DQ alpha, DP beta, DQ beta and DR beta probes, about 20 polymorphic bands being detected for each. DR alpha showed 2-4 polymorphic bands, whereas no evidence for DP alpha-like genes was found. A number of correlations of RFLPs with individual alloantisera were apparent.
Raeside JI, Christie HL.C(18) neutral steroid formation by cytochrome P450 aromatase has been recorded for several equine and porcine tissues. High activity of P450 aromatase is reflected in the quantities of estrogens in yolk-sac (y-s) fluid of early equine conceptuses. In a previous study of y-s fluid we detected large amounts of androgens by radioimmunoassay (RIA), using an antiserum for androstenedione (A(4)). Here, we report that RIA, following chromatography, gave tentative identification of the major peak as norandrostenedione (19-norA) not as A(4). Furthermore, even greater quantities of 19-norA seemed to be ...
Kwok WH, Kwok KY, Leung DK, Leung GN, Wong CH, Wong JK, Wan TS.The positive findings of norbolethone in 2002 and tetrahydrogestrinone in 2003 in human athlete samples confirmed that designer steroids were indeed being abused in human sports. In 2005, an addition to the family of designer steroids called 'Madol' [also known as desoxy-methyltestosterone (DMT)] was seized by government officials at the US-Canadian border. Two years later, a positive finding of DMT was reported in a mixed martial arts athlete's sample. It is not uncommon that doping agents used in human sports would likewise be abused in equine sports. Designer steroids would, therefore, pose...
Bueno AC, Galuppo LD, Taylor KT, Jensen DG, Stover SM.To compare the mechanical shear strengths and stiffnesses obtained from in vitro testing of a simulated complete third carpal bone (C3) frontal plane radial facet slab fracture (osteotomy) stabilized with either a 4/5 Acutrak (AT) compression screw or a 4.5-mm AO cortical bone (AO) screw inserted in lag fashion. Drilling, tapping, and screw insertion torques, forces, and times also were compared between AT and AO implants. Methods: In vitro biomechanical assessment of site preparation, screw insertion, and shear failure test variables of bone screw stabilized simulated C3 slab fracture in pair...
Salloum RM, Duckworth D, Madison JB, Souba WW.The sodium-dependent transporter system responsible for L-glutamine uptake by brush border membrane vesicles prepared from equine jejunum was characterized. Vesicle purity was ascertained by a 14- to 17-fold increase in activity of the brush border enzyme markers. Glutamine uptake was found to occur into an osmotically active space with negligible membrane binding. The sodium-dependent velocity represented approximately 80% of total uptake and demonstrated overshoots. Kinetic studies of sodium-dependent glutamine transport at concentrations between 5 microM and 5 mM revealed a single saturable...
Turek JJ, Lamar CH, Fessler JF, Bottoms GD.An in vitro system of cultured equine endothelial cells was evaluated as a model for endotoxin (ET) exposure in the horse. Primary cell lines from pulmonary vessels and aortas were cultured from tissues of 6 horses. Effects of ET alone with and without serum and in combination with the cyclo-oxygenase inhibitor flunixin meglumine and isolated equine neutrophils were evaluated by transmission electron microscopy. Cells plus serum were incubated with 10, 25, 50, or 100 micrograms of ET/ml of incubation medium for 1, 3, 8, or 24 hours. Cells without serum were cultured for 1 and 3 hours. Flunixin...
Leclere M, Chevalier Z, Dubuc V, Beauchamp G, Bédard C.The amount of thrombin generated reflects the endogenous thrombin potential (ETP), which depends on the balance of pro- and anticoagulant factors. The calibrated automated thrombogram (CAT) allows for the direct measurement of thrombin generation during the clotting process. Objective: (1) To describe the results of the CAT assay in horses, (2) to establish intra-assay and intra- and interindividual variation of thrombin generation in healthy horses, and (3) to compare in vitro low-molecular-weight heparin (LMWH) sensitivity between healthy and sick horses. The hypothesis for the last objectiv...
Rosendal S, Lumsden JH, Viel L, Physick-Sheard PW.Neutrophils were isolated from the peripheral blood of adult equids (group 1) and were purified on a density gradient of polyvinylpyrrolidone-coated silica gel. A bactericidal assay was developed, using an equine skin isolate of Staphylococcus epidermidis as target bacterium in medium containing pooled fresh equine serum for opsonization. Significant (P less than 0.05) killing was observed after 60 or 120 minutes' incubation. Reduction in bactericidal function of blood neutrophils was not found after incubation with a virulent strain of Mycoplasma felis for 30 or 60 minutes. Similarly, the fun...
Quinteros DD, García-López JM, Abuja GA, Tarkanian M, Maranda LS, Bubeck K, Kowaleski MP.To determine if the mechanical properties (strength and stiffness) of a new prototype 4.5 mm broad locking plate (NP-LP) are comparable with those of a traditional 4.5 mm broad limited-contact self compression plate (LC-SCP), and to compare the bending and torsional properties of the NP-LP and LC-SCP when used in osteotomized equine third metacarpal bones (MC3). Methods: The plates alone were tested in four-point bending single cycle to failure. The MC3-plate constructs were created with mid-diaphyseal osteotomies with a 1 cm gap. Constructs were tested in four-point bending single cycle to fa...
Kirschvink N, Art T, Lekeux P, Roberts C, Gustin P.8-Epi-PGF2alpha, a prostaglandin-like compound generated by oxidative stress, has been shown to be an in vitro bronchoconstrictor in airways from healthy laboratory animals and healthy humans, but it has never been studied in diseased airways. Here, the bronchoconstrictive capacity of 8-epi-PGF2alpha on isolated bronchial rings (BR) of healthy and heaves-affected horses was evaluated by comparing the maximal effect and the potency of 8-epi-PGF2alpha to those of (1) acetylcholine (ACh), (2) its stereoisomer PGF2alpha and (3) its synthetic receptor agonist, U46619. Furthermore, the potential cap...
Weston MC, Collins ME, Cunningham FM.Inappropriately activated eosinophils can contribute to disease pathogenesis and intracellular signalling pathways that regulate functional responses may represent a therapeutic target. Little is known about intracellular signalling in equine eosinophils and this study examined the role of phospholipase C (PLC) and a range of protein kinases on responses to histamine and CCL11. Histamine (10(-4) M) or CCL11 (5.6 x 10(-9) M)-induced actin polymerization, migration and superoxide production by eosinophils from healthy horses were compared in the presence and absence of selective kinase inhibitor...
Wilsher S, Rigali F, Kovacsy S, Allen WT.Vitrification of embryos >300 µm in diameter requires puncture of the glycoprotein capsule, although the size of the hole compatible with embryo survival is unknown. Forty-five day-7 or -8 embryos were punctured using a 30-µm glass biopsy pipette mounted on a micromanipulator (n = 20) or manually with either an acupuncture needle (∼100-µm diameter -hole; n = 10) or a microneedle with a <1 µm tip to produce a ∼30-µm diameter hole (n = 15) before transferring to recipient mares; further 12 embryos were punctured with either the acupuncture needle or microneedle before being cultured ...
Major MD, Grant BD, White KK, Ratzlaff MH, Gallina AM, Crawley GR.Mechanical properties of equine suspensory apparatus preparations and three braided synthetic prostheses were evaluated in vitro. Force versus displacement plots and failure modes were recorded from single load-to-failure testing in 18 cadaver limbs before and after replacement of each suspensory apparatus with a prosthesis. Mean load at failure, energy to failure, and stiffness values of polytetrafluoroethylene (PTFE) prostheses were lower than those of the suspensory apparatus and aramid prosthesis. The PTFE prosthesis failed by elongation or rupture of the prosthesis. Mechanical properties ...
Hillman RB, Olar TT, Squires EL, Pickett BW.Stallion semen was collected, using artificial vaginas at 44 to 46, 48 to 50, and 52 to 54 C, to study the effects of temperature on seminal quality and sexual behavior. The temperature of the artificial vagina had no significant effect on motility, gel volume, gel-free seminal volume, total seminal volume, pH, number of mounts per ejaculate, total time to ejaculation, or seminal temperature. Spermatozoa were collected, then exposed to water-bath temperatures of 38, 45, 49, or 53 C for 1 minute. Mean motility was similar after exposure to temperatures of 38 or 45 C, but exposure to temperature...
