Laboratory methods in equine research encompass a variety of techniques and procedures used to analyze biological samples from horses to study health, disease, genetics, and physiology. These methods include hematological analyses, biochemical assays, molecular biology techniques, and microbiological cultures. Commonly utilized laboratory techniques involve blood tests for complete blood count (CBC) and serum chemistry, polymerase chain reaction (PCR) for genetic and infectious disease studies, and enzyme-linked immunosorbent assays (ELISA) for detecting specific proteins or antibodies. These methods provide valuable data that contribute to understanding equine health and disease mechanisms. This page compiles peer-reviewed research studies and scholarly articles that explore the application, development, and outcomes of laboratory methods in the context of equine research.
Courtot D.At the request of the Service des Haras, our laboratory works on the toxicological problems of the sport-horse. These studies have resulted in the setting up of an anti-doping control for equestrian competitions of various types, not only flat racing. During events, horses, must be calm and docile to the riders' order. Frequently, the latter use tranquillizers to try and win events. The analytical method for the research and identification of these compounds is described. The technique involves successively: 1. alkalinisation of the sample - saliva, blood or urine after enzymatic hydrolysis. 2...
Haywood PE, Chalmers P.The chromatographic and spectroscopic properties of several unusual substances which have been detected in the "alkaloidal" chloroform extract from racehorse urine and saliva samples are reported. Some of these substances have been identified by combined gas chromatography-mass spectrometry and the source of the substance is stated where this is known. Other substances whose identity is not known have been detected and their mass spectra show characteristic amine fragments. The occurrence of these unidentified substances is more frequent in aged urine samples and it would therefore appear that...
Nakajima T, Matsumoto T.Horse racing in Japan consists of two systems, the National (10 racecourses) and the Regional public racing (32 racecourses) having about 2,500 racing meetings in total per year. Urine or saliva samples for dope testing are collected by the officials from thw winner, second and third, and transported to the laboratory in a frozen state. In 1975, 76, 117 samples were analyzed by this laboratory. The laboratory provides the following four methods of analysis, which are variously combined by request. (1) Method for detection of drugs extracted by chloroform from alkalinized sample. (2) Methods fo...
Kundriutskova LA, Kruglikova RI.Hydrolysis of ethers of saturated and unsaturated alcohols and ethers, e.g. phenol and choline, under the action of horse blood serum cholinesterase, was studied. The reactivity towards enzymatic hydrolysis is decreased due to a greater length of the chain in the alcohol residue of the benzoic acid aminoethers; at nCH2 = 4 the compound is a poor substrate. An increase in nydrophobicity of the acyl residue of the ether molecule also leads to a decrease in the Vmax and Km values. In case of cholinesterase substrates, an increase in the molecule hydrophobicity results in an increase of its non-pr...
Helmstaedter V, Feurle GE, Forssmann WG.Equine pancreas was investigated with immunohistochemical methods to study the distribution of endocrine cells immunoreactive to anti-insulin, anti-glucagon, and anti-somatostatin. A-cells demonstrable by anti-glucagon are located in the center of Langerhans islets and frequently in the duct epithelium. Few A-cells are seen associated to acini. Anti-insulin reactive B-cells form a large zone around the center of the Langerhans islets in which some B-cells lie between exocrine cells and others, although few, are located in the duct epithelium. D-cells stained with anti-somatostatin serum form a...
Wilbur DJ, Allerhand A.The titration behavior of individual tyrosine residues of myoglobins has been studied by observing the pH dependence of the chemical shifts of Czeta and Cgamma of these residues in natural abundance of 13C Fourier transform NMR spectra (at 15.18 MHz, in 20-mm sample tubes, at 37 degrees) of cyanoferrimyoglobins from sperm whale, horse, and red kangaroo. A comparison of the pH dependence of the spectra of the three proteins yielded specific assignments for the resonance of Tyr-151 (sperm whale) and Tyr-103 (sperm whale and horse). Selective proton decoupling yielded specific assignments for Cze...
Glöckner WM, Newman RA, Dahr W, Uhlenbruck G.Phenol extraction of horse, sheep, cow, pig and human erythrocyte membranes and human milk fat globule membranes gave glycoprotein fractions, all of which were shown by gas chromatography to contain the reduced disaccharide beta-D-galactosyl (1-3)-N-acetyl-D-galactosaminital after treatment with alkaline borohydride. Cow and pig erythrocyte membrane glycoproteins were found however to contain much lower amounts than the erythrocyte membrane glycoproteins of the other species tested. After gel filtration, a tetrasaccharide was isolated from horse and sheep glycoproteins containing the disacchar...
Rance TA, Park BK, Rowe PH, Dean PD.A radioimmunoassay procedure was developed for the measurement of dehydroepiandrosterone (DHA) in peripheral serum in nonpregnant mares. The synthesis and conjugation of 3beta-hydroxy-5-androsten-19-al-17-one 19(0-carboxymethyl) oxime is described. Antisera were developed against this antigen and characterized. The most specific antiserum was used to measure DHA. Concentrations of DHA were greatest immediately before ovulation.
Sampaio MU, Galembeck F, Paiva AC, Prado ES.The kinetic constants for horse urinary kallikrein and trypsin hydrolysis of BAEE, TAME, bradykinin methyl ester and bradykinyl-Ser-Val-Gin-Val-Ser were determined. The values of the ratio kcat/Km show that (1) kallikrein is catalytically less efficient than trypsin for all the substrates (2) the three esters are equally good substrates for trypsin while horse urinary kallikrein is 100-fold more effective on bradykinin methyl ester than on the other substrates (3) for both enzymes the ester of bradykinin is a better substrate than the tetradecapeptide.
Sampaio CA, Prado ES.Ala-Phe-Lys-CH2-Cl is a chloromethylketone derivative which is able to promote the inhibition of several proteolytic enzymes. In this paper the inhibition of horse urinary and plasmatic kallikreins is described and this inhibition is compared to that produced in human plasma kallikrein. This compound was designed based upon the structure of bradykinin. This enzyme substrate system can provide a model for the study of the interactions between bradykinin and its receptor. The inhibition of the enzymes was achieved both for its esterase and kinin-releasing activities.
Holowka DA, Cathou RE.IgM antibodies specific for the fluorophore epsiolon-1-dimethylamino-5-naphthalenesulfonyl-L-lysine(DNS-lysine) were elicited in the horse and nurse shark by immunization with a DNS-lysine streptococcal conjugate; the antibodies were purified by specific adsorption with an immunoadsorbent followed by gel filtration to select the IgM class (molecular weight 900 000). About 90% of the equine anti-DNS was IgM.DNS-Lysine, when bound in the combining sites of a population of these anti-DNS IgM antigodies from horse and nurse shark, as well as from pig, exhibited a marked fluorescence enhancement an...
Saiatov MKh, Beĭsembaeva RU.Profiles of distribution of non-specific gamma-inhibitors of influenza A2/Victoria/35/72 in donkey and horse sera were established by gel chromatography in Sephadex G-200. High and low molecular inhibitors were found in 19S and 4S serum fractions. Highly purified preparations of a2-macroglobulin, transferrine and albumin were isolated by a combination of methods of salt precipitation, gel chromatography on Sephadex G-100, G-200 and ion exchange on DEAE-Sephadex A-50. Heating sera resulted in a considerable increase of the antiviral activity of a2-macroglobulin and transferrine and a reduction ...
Korkeala H, Stabel-Taucher R, Pekkanen TJ.When 33 horse kidneys were tested for the presence of inhibitory substances by the Bacillus subtilis BGA method at pH 8 and the Micrococcus luteus ATCC 9341 method, 24 were positive and 9 negative. The pH of the seeded M. luteus test medium changed from pH 6.6 before incubation to 8.7 after 24 hours incubation at 30 degrees C. When the same 33 kidneys were tested by the B. subtilis BGA method, medium pH 6, and 15 of them also by the M. luteus method using a medium buffered to pH 6, all were negative. The cadmium concentration of the 33 horse kidneys was found to be 70.17 +/- 81.28 mg/kg wet we...
Targowski SP.The present study describes a two step technique for the separation of mononuclear leukocytes or mononuclear and polymorphonuclear leukocytes from whole equine blood. First, the leukocyte rich plasma was obtained by sedimentation of erythrocytes in the undiluted blood. Subsequently, separation of the different populations of white blood cells was performed by centrifugation with different gradients overlaid with the leukocyte rich plasma. The optimal separation of the mononuclear cells was obtained by the centrifugation of the leukocyte rich plasma overlaying the gradient containing 24 parts o...
Johnson PE, Maister SG, Knowles JR.Phosphoglycerate kinase has been isolated in crystalline form from horse muscle. A convenient isolation procedure is described that yields homogeneous enzyme of specific activity 700 units/mg (30 degrees C). The enzyme is monomeric, and has a molecular weight 47 000. Of the eight cysteine residues in the protein, two react rapidly with Nbs21 with the concomitant loss of the catalytic activity. Since the isolation of phosphoglycerate kinase from yeast (Bücher, 1955) there have been several reports of purification methods yielding enzyme approaching molecular homogeneity, from rabbit muscle (Be...
Scherer WF, Anderson K, Pancake BA, Dickerman RW, Ordonez JV.Seventy-four strains of Venezuelan encephalitis (VE) virus recovered from sentinel hamsters or mosquitoes at enzootic habitats in Guatemala in the two years following the 1969 epidemic-equine epizootic were examined for ability to produce small plaques in Vero African green monkey kidney cell cultures, like isolates obtained during the epizootic. (a) One strain recovered from a sentinel hamster in late October 1969 at an enzootic habitat near the epicenter of the hemagglutination-inhibition (HI) and equine-virulence properties like epizootic virus; this strain retained its small plaque charact...
Poskus E, Zakin MM, Fernámdez HN, Paladini AC.Peptide fragments, obtained from equine growth hormone by cyanogen bromide cleavage and further chemical treatment, were isolated and identified. Their immunological reactivities were tested by hemagglutination and complement fixation methods using rabbit antisera against native hormone. Antigenic determinants were detected in the fragments comprising amino acid sequences 5-72 and 73-123, this last one being predominant. Fragment 124-178 had very low reactivity. Nitration of peptide 73-123 did not modify its immunological properties,but oxidation diminished them. Comparison of the antigenicity...
Ronda GJ, Gaastra W, Beintema JJ.The kinetic parameters Km, k+2 and k+2/Km of the pancreatic ribonucleases (EC 3.1.4.22) from cow, giraffe, horse, rat and lesser rorqual have been determined, using 2',3'-cyclic cytidine monophosphate and 2',3'-cuclic uridine monophosphate as substrates. No large differences were found between the activities of the five enzymes. The relative differences between the activities of the five enzymes are mainly due to differences in the rates of hydrolysis and not to differences in the affinities for the substrates.
Kotts C, Jenness R.Kappa-Casein-like proteins were isolated from the milks of cow, goat, reindeer, horse, rat, and rabbit. When treated with rennin, all of the isolated kappa-casein components yielded para-kappa-casein-like bands on gel electrophoresis. The rate of cleavage of these components with rennin was determined by measuring material soluble in trichloroacetic acid (macropeptide). The curves were characteristic of a limited, specific attack by rennin on these proteins. The goat and reindeer kappa-caseins were nearly as bovine kappa-casein, but the cleavage of horse, rat, and rabbit kappa-casein-like comp...
Kihurani DO, Masake RA, Nantulya VM, Mbiuki SM.Following an outbreak of trypanosomosis in horses on a farm in Kenya, 18 trypanosome isolates were collected from the infected animals over a period of one and a half years and cryopreserved for characterization. The characterization was done on the basis of morphology using Giemsa-stained blood and buffy coat smears, infectivity to mice, recombinant DNA hybridization, and chromosome separation by orthogonal field alternation gel electrophoresis (OFAGE). Morphologically, all the trypanosome isolates were identified as belonging to the subgenus Nannomonas, and a total of 16 out of the 18 isolat...
De Ceulaer K, Van Ginneken C, Delesalle C, Van Brantegem L, Deprez P, Weyns A.This study aimed to evaluate the reliability of slaughterhouse-obtained small intestinal tissue as control material in equine colic research where molecular stress responses in small intestinal tissue are investigated. For this purpose, small intestinal samples from colic horses were collected during surgery or immediately after euthanasia at the oral border of strangulation resection sites and routinely processed for histopathology (i.c. rinsed with 4°C Krebs' solution, fixated overnight with 4% neutral buffered formaldehyde (FH) at room temperature). Control samples consisted of pieces of m...
Meistrich ML.The problem of extrapolating effects of reproductive toxins on experimental animals to predict the doses that would produce infertility in human males is discussed using published data on effects of testosterone and estradiol on sperm production in the rat, rabbit, rhesus monkey, ram, stallion, and humans. This analysis indicates that calculation of the dose of testosterone that reduces human sperm counts by a given percentage is best done using the dose administered to laboratory animals expressed on the basis of body weight, as opposed to some other parameter such as body surface area. A sur...
Stott ML, Osburn BI.Long-term equine lymphocyte cultures were initiated and maintained in continuous culture with medium containing recombinant human interleukin-2. Cultures were successfully maintained with lectin activation signals and recombinant human interleukin-2 or with recombinant human interleukin-2 alone. All cell cultures that were characterized had a T-lymphocyte phenotype and had lectin-dependent or -independent cytotoxicity directed to various cell types. These findings demonstrate that long-term equine T-lymphocytes cultures can be initiated and maintained easily.
Catena C, Asprea L, Carta S, Tortora G, Conti D, Parasacchi P, Righi E.We have investigated and compared DNA damage and cell killing induced in human and equine lymphocytes after in vitro X-irradiation. Our data show that the cytogenetic and the lethality effects are both greater in equine lymphocytes, but that the difference is wider for lethality. The ratios between doses inducing the same effect are 1.3, 1.7 and 9.4 for the number of binucleated cells with micronuclei, micronucleus frequency in binucleated cells and DNA synthesis inhibition, respectively. The very different radiosensitivity observed for the two mammalian species encourages us to use their lymp...
Freer H, Hillegas JM, Wimer C, Baldwin C, LaBresh J, Wagner B.Interleukin-2 (IL-2) is a T cell growth factor and major modulator of T helper (Th) cell differentiation. Here, we have developed and characterized a monoclonal antibody to equine IL-2 (anti-IL-2 mAb, clone 158-1). The IL-2 mAb detected rIL-2 by ELISA, intracellular staining and flow cytometry analysis and Western blotting. The IL-2 mAb was also paired with a polyclonal IL-2 detection antibody in both ELISA and a fluorescent bead-based assay. When these two assays were compared using identical reagents there was an improved analytical sensitivity (46pg/ml) and wider linear quantification range...
Gregg AS, Jones RS, Snowdon SL.A fault in the assembly of a Matrix Large Animal Circle anaesthetic machine resulted in reversal of fresh gas flow through the vaporizer. The fault was discovered only after the sudden development of excessive depth of anaesthesia in two equine patients. Laboratory investigations were conducted to determine the effect of flow reversal on vaporizer output. Results indicated that output concentration was approximately doubled under these conditions.
Suter M, Fey H.Horse IgE was isolated from a serum pool collected from foals naturally infected with endoparasites. The serum was precipitated with ammonium sulfate, delipidated with dextran sulfate and further purified by gel filtration, anionic exchange, immunosorption or preparative polyacrylamide gelelectrophoresis. By these methods IgE could be isolated at a purity of 81%. The sera from rabbits immunized with the purified horse serum fractions were tested using reversed passive cutaneous anaphylaxis and an enzyme linked immunosorbent assay (ELISA). By the ELISA method cross reaction of rabbit anti horse...
Fischer A, Lechner J, Kraft W, Hirschberger J.The QBC is a centrifugal haematology system. Modified haematocrit capillaries are measured optically. The parameters haematocrit, total leukocyte count, relative and absolute values of granulocyte and lympho-/monocyte fractions, and total thrombocyte count are stated. The microhaematocrit method, the counting chamber, and the differential blood count are reference methods. Blood samples of the dog, cat and horse were used for the study. As a screening method the QBC analysis meets all requirements for the veterinary practice.
Leung GN, Tang FP, Wan TS, Wong CH, Lam KK, Stewart BD.This paper describes the application of gas chromatography-mass spectrometry (GC-MS) for in vitro and in vivo studies of 6-OXO in horses, with a special aim to identify the most appropriate target metabolite to be monitored for controlling the administration of 6-OXO in racehorses. In vitro studies of 6-OXO were performed using horse liver microsomes. The major biotransformation observed was reduction of one keto group at the C3 or C6 positions. Three in vitro metabolites, namely 6alpha-hydroxyandrost-4-ene-3,17-dione (M1), 3alpha-hydroxyandrost-4-ene-6,17-dione (M2a) and 3beta-hydroxyandrost-...
Pedroso RC, Salvadori MC, Andraus MH, Lopez NM.A chromatographic method was developed to detect and confirm the presence of succinonitrile (SDN) in horse urine samples, for antidoping control. The urine samples (5 ml) were extracted with diethyl ether and screened by gas chromatography-nitrogen-phosphorus detector and the confirmation of the drug's presence was accomplished by using gas chromatography-mass selective detection. The recovery of extraction was 78 and 81% for 1.0 and 2.0 micrograms ml-1 (relative standard deviation, < 10%), respectively. Urine samples collected after the administration of Energisan were positive for SDN (1-30 ...
Sharma OP.The concentrations of PGF-2alpha in the peripheral blood of five foaling mares were measured by radioimmunoassay. Low levels of PGF-2alpha were detected as early as 1 week before foaling in two of the mares. These levels increased steadily, reaching a peak (1-74 +/- 0-44 ng/ml) during fetal expulsion. A relatively high PGF-2alpha level was found in samples collected 60 min after foaling.
Johansson IM, Schubert B.Flunixin is determined in equine plasma by liquid chromatography on LiChrosorb RP-18 with 70% methanol in phosphate buffer pH 3.1 as the eluent, with detection at 284 nm. Plasma is deproteinized with methanol and the supernatant is then injected directly into the system. With a short pre-column (5 x 3 mm i.d.), which is replaced after 25-40 injections of sample, 420 plasma samples could be analysed on one analytical column. The detection limit in plasma is 0.30 micromol/l (89 ng/ml) and the method can be used in pharmacokinetic studies.
Rozanova SL, Rozanova ED, Nardid OA.Experimental data are presented which were obtained under comparative evaluation of influence of different freezing-thawing conditions on antioxidant properties of isolated proteins: human serum albumin, cytochrome c from the horse heart and glucose oxidase from Aspergillus niger. The observed protein antioxidant activity alterations are assumed to be a result of protein conformational changes. The character of freezing-thawing influence on the protein antioxidant activity depends on the molecular structure and cooling conditions.
Ledwozyw A, Jabłonka S, Tusińska E, Herbut M.The levels of endogenous heparin in the plasmas of horses, cows, sheep and pigs were determined with the use of synthetic chromogenic substrate benzoyl-isoleucyl-glutamyl-glycyl-arginyl-p-nitroanilide (S-2222). The lowest heparin concentrations were stated in cattle plasma, the highest ones in the plasma of pigs.
McCoy DJ, Beasley R.Hypercalcemia associated with malignancy was diagnosed in a 2-year-old Thoroughbred filly admitted because of weight loss and reduced exercise tolerance of approximately 2 months' duration. Laboratory findings included hypercalcemia, hypophosphatemia, anemia, marked neutrophilia with lymphopenia and eosinopenia, and normal immunoreactive parathyroid hormone concentration. At necropsy, a 53.6-kg tumor was located in the cranioventral aspect of the abdominal cavity. Gross renal lesions were not noticed. Bone tissue appeared to be normal on gross and histologic examinations. The parathyroid gland...
Magnuson NS, Perryman LE, Decker DM, Magnuson JA.2'-Deoxyadenosine and 9-beta-D-arabinofuranosyladenine (ARA) are apparent suicide inhibitors for equine S-adenosylhomocysteine hydrolase. In initial velocity studies of the synthetic reaction converting adenosine and homocysteine to S-adenosylhomocysteine, adenine, adenosine 5'-triphosphate, and 9-beta-D-arabinofuranosyladenine were found to be competitive inhibitors with Kis of 3.8 microM, 1.1 mM, and 30 microM, respectively. In contrast, linear mixed inhibition was observed for 2'-deoxyadenosine, indicating that 2'-deoxyadenosine must bind in more than one fashion to the enzyme.
Rivero JL, Cascone O, Biscoglio de Jimenez Bonino MJ.1. The method of Kubota et al. [Biochim. biophys. Acta 701, 242-252 (1982)] was applied to several members of the growth hormone family in order to examine their conformational homology. 2. The method neither detects differences between rat, cow, sheep, horse and alpaca hormones, nor between monkey and human hormones. 3. Lack of homology between primate and non-primate growth hormones was found in segments 42-49 and 184-191. The first fragment could be linked to species-specificity.
Qureshi GA, Eriksson A.A method for the routine determination of the beta-adrenergic drugs clenbuterol and mabuterol in equine plasma has been developed. The drugs were isolated from alkalinized plasma by liquid-liquid extraction. The organic phase was evaporated to dryness and the residue was dissolved in the mobile phase prior to injection. The recoveries were 98% and 95% for clenbuterol and mabuterol, respectively. The drugs were separated by reversed-phase high-performance liquid chromatography and quantitated by a use of a coulometric detector set at +0.75 V vs. the internal reference electrode. The influence o...
Bugno M, Jabłońska Z, Słota E.The aim of the study was to optimize hybridization conditions of molecular probes specific for X sex chromosomes of the domestic horse in mare oocyte chromosomes. Mare oocytes, recovered from slaughterhouse ovaries by scraping the granulosa layer, were cultured in vitro. Metaphase II mature oocytes were treated with hypotonic solution and fixed, followed by hybridization of the molecular probe specific for the X chromosome ofthe domestic horse. Hybridization of probes specific for mouse heterosomes on mouse oocytes and early embryos was performed to verify the FISH technique. Of 438 oocytes an...
Higashi T, Harasawa R.The three equine adenovirus strains isolated in different locations showed a similar cleavage pattern with HindIII and the DNA homology among the strains was confirmed by Southern blot hybridization. The three strains revealed differences in cleavage patterns with BamHI, EcoRI and PstI, suggesting the presence of DNA polymorphisms among equine adenoviruses.
Stalheim OH, Cottew GS, Freundt EA, Koski TA, Leach RH, Perreau P, Stone SS.Antisera to 10 mycoplasma species of bovine origin were produced in 10 ponies and were distributed for evaluation in growth-inhibition tests at 6 laboratories in Australia, England, Denmark, France, and the United States. Except for a few failures with some antigens produced at the 6 laboratories, the antisera induced large zones of growth inhibition in homologous, but not heterologous, systems. These antisera may be useful as standard reagents for the identification of the bovine mycoplasmas.
Saint-Dizier M, Foulon-Gauze F, Lecompte F, Combarnous Y, Chopineau M.In order to investigate the role of the unique seventh N23-glycosylation site of the equine LH/CG receptor (eLHCGR) in the cAMP pathway activation, COS-7 cells were transiently transfected with either the wild-type or the mutant eLHCGR(N23Q) cDNA and challenged with porcine LH and eCG for cAMP production. We showed that the N23-glycosylation site of the eLHCGR is not required for the functional coupling of the receptor with the cAMP pathway and is not responsible for the limited potency of eCG relative to pLH to activate this receptor.
Bacigalupo MA, Ius A, Meroni G, Dovis M, Petruzzelli E.A time-resolved fluoroimmunoassay (TR-FIA) for the direct determination of clenbuterol residues in horse urine using a highly specific monoclonal antibody has been compared with an immunoenzymometric assay (IEMA). The sensitivity of both methods was 10 pg; the calibration curve was linear between 10 and 10(5) pg for the TR-FIA and between 10 and 10(4) pg for the IEMA.
Allen BV.A technique for automating equine differential leucocyte counts by analysis of volume distribution curves using the Coulter Channelyzer has been developed and evaluated. A comparison between the results obtained by this method and standard microscopic techniques showed good agreement in most cases. Blood samples can be analysed for both differential and total leucocyte counts at a rate of 25/h. For each sample an average 16,000 leucocytes are classified by the Channelyzer. The method of volume analysis is suitable for the precise counting of polymorphonuclear neutrophils, lymphocytes and eosin...
Takesue M, Osaka Y, Muranaka M, Katayama Y, Ikadai H.In the present study, the serum and cerebrospinal fluid of horses diagnosed with Setaria digitata cerebrospinal filariasis were analyzed by western blot. The results revealed S. digitata protein bands measuring 65, 34, 22, and 18 kDa in molecular weight. In particular, the 18 kDa band is a possible candidate for clinical immunodiagnosis on the basis of western blot findings.
Simpson KS, Adams MH, Behrendt-Adam CY, Baker CB, McDowell KJ.Complex changes in gene expression must occur at the proper time and in the appropriate tissues for pregnancy to be successful. Therefore, research aimed at defining the regulation of gene expression in conceptuses is of critical importance. However, information on developmentally regulated changes in gene expression in horse conceptuses is sparse and inadequate. In the present study, suppression subtractive hybridization was used to identify genes that are expressed more highly at day 15 than on day 12 of gestation. This period encompasses maternal recognition of pregnancy and the beginning o...
Brostroöm H, Paulie S, Perlmann P.To characterise the expression of surface antigens on equine sarcoid cells compared to normal equine fibroblasts, immune sera were produced in rabbits against transformed cells of a virus-containing sarcoid cell line (Mc-1) and normal dermal fibroblasts, respectively. The specificities of the sera were analysed by antibody-dependent cellular cytotoxicity against 51Cr-labelled target cells using human lymphocytes as effector cells. Anti-Mc-1 antiserum induced strong cytotoxicity against transformed cells of two sarcoid cell lines (Mc-1 and Bay Mc-1), whereas the cytotoxicity against transformed...
Murata T, Yamashiro Y, Kondo T, Nakaichi M, Une S, Taura Y.Complementary DNA (cDNA) for bovine quaking gene (Bqk), equine quaking gene (Eqk) and porcine quaking gene (Pqk), which are homologous to mouse quaking gene (qkI), were isolated, and their nucleotide sequences were determined. cDNA sequences of Bqk, Eqk and Pqk showed very high homology to that of qkI at nucleotide level; 94.2, 95.7 and 95.6%, respectively. Deduced amino acid sequences for Bqk, Eqk and Pqk perfectly matched to that of qkI. These findings suggest that the quaking gene family is highly conserved during mammalian evolution, and that Bqk, Eqk and Pqk are likely to have important b...
Boudouard M, Giudicelli J, Sudaka P.A rapid method for preparation of brush border membrane vesicles from a large amount of horse kidney cortex is described. Self-orienting Percoll-gradient centrifugation minimized contamination by microsomal membranes. The characteristics of this preparation were checked by electron microscopy and measurement of L-alanine uptake.
