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Topic:In Vitro Research

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.
In vitro effects of bethanechol on equine gastrointestinal contractility and functional characterization of involved muscarinic receptor subtypes.
Journal of veterinary pharmacology and therapeutics    December 14, 2005   Volume 28, Issue 6 565-574 doi: 10.1111/j.1365-2885.2005.00693.x
Marti M, Mevissen M, Althaus H, Steiner A.The goal of this study was to investigate the effect of bethanechol (BeCh) on contractility patterns of smooth muscle preparations of equine duodenum descendens, jejunum, caecum and pelvic flexure in vitro. Concentration-response relationships were developed for BeCh using in vitro assays with and without preincubation of muscarinic (M) receptor antagonists for M2 and M3 receptors. BeCh induced a significant, concentration-dependent increase in contractile response in equine intestine in specimens with circular orientation. The maximal effect was largest for jejunal specimens with no differenc...
An in vitro evaluation of plate luting using osteotomized equine third metacarpal bones with a limited contact-dynamic compression plate.
Veterinary surgery : VS    December 14, 2005   Volume 34, Issue 6 587-593 doi: 10.1111/j.1532-950X.2005.00091.x
Sod GA, Hubert JD, Martin GS, Gill MS.To evaluate the effects of plate luting on the biomechanical properties of a broad limited contact-dynamic compression plate (LC-DCP) fixation to repair osteotomized equine 3rd metacarpal (MC3) bones. Methods: In vitro biomechanical testing of paired cadaveric equine MC3 with a mid-diaphyseal osteotomy, stabilized by LC-DCP fixation, with 1 of the pair luted with polymethylmethacrylate (PMMA). Methods: Ten pairs of adult equine cadaveric MC3 bones. Methods: Ten pairs of equine MC3 were divided into 2 test groups (5 pairs each) for (1) palmarodorsal 4-point bending single cycle to failure testi...
Effects of induction of capacitative calcium entry on equine laminar microvessels.
American journal of veterinary research    December 13, 2005   Volume 66, Issue 11 1877-1880 doi: 10.2460/ajvr.2005.66.1877
Robertson TP, Peroni JF, Lewis SJ, Moore JN.To determine the effects of induction of capacitative Ca2+ entry on tone in equine laminar arteries and veins. Methods: Laminar arteries and veins from 6 adult mixed-breed horses. Methods: Arteries and veins were isolated and mounted on small vessel myographs for the measurement of isometric tension. Capacitative Ca2+ entry was induced by incubating the vessels with the specific Ca2+-ATPase inhibitor thapsigargin (100nM) in a Ca2+-free physiologic salt solution. Capacitative Ca2+ entry-associated contractile responses were determined by the subsequent addition of 2mM Ca2+ to the solution bathi...
Effects of glucosamine and chondroitin sulfate on mediators of osteoarthritis in cultured equine chondrocytes stimulated by use of recombinant equine interleukin-1beta.
American journal of veterinary research    December 13, 2005   Volume 66, Issue 11 1861-1869 doi: 10.2460/ajvr.2005.66.1861
Neil KM, Orth MW, Coussens PM, Chan PS, Caron JP.To determine whether glucosamine and chondroitin sulfate (CS) at concentrations approximating those achieved in plasma by oral administration would influence gene expression of selected mediators of osteoarthritis in cytokine-stimulated equine articular chondrocytes. Methods: Samples of grossly normal articular cartilage obtained from the metacarpophalangeal joint of 13 horses. Methods: Equine chondrocytes in pellet culture were stimulated with a subsaturating dose of recombinant equine interleukin (reIL)-1beta. Effects of prior incubation with glucosamine (2.5 to 10.0 microg/mL) and CS (5.0 t...
Evaluation of coexpression of microsomal prostaglandin E synthase-1 and cyclooxygenase-2 in interleukin-1-stimulated equine articular chondrocytes.
American journal of veterinary research    December 13, 2005   Volume 66, Issue 11 1985-1991 doi: 10.2460/ajvr.2005.66.1985
Farley J, Sirois J, MacFarlane PH, Kombé A, Laverty S.To characterize expression of cyclooxygenase-2 (COX-2) and microsomal prostaglandin E synthase-1 (mPGES-1) and regulation of prostaglandin E2 (PGE2) production by equine articular chondrocytes. Methods: Articular cartilage from the metacarpophalangeal joints of 7 adult horses. Methods: Equine chondrocyte monolayer cultures were stimulated with different concentrations (2.5, 5, 10, and 20 ng/mL) of recombinant human interleukin-1beta (rhIL-1beta) for 24 hours and then with rhIL-1beta (5 ng/mL) for 3, 6, 9, 12, and 24 hours. Concentration of PGE2 in the media was measured via radioimmunoassay. T...
Efficacy demonstration of tetanus vaccines by double antigen ELISA.
Pharmeuropa bio    December 13, 2005   Volume 2005, Issue 1 31-52 
Rosskopf U, Noeske K, Werner E.This paper describes a double antigen ELISA (DAE) for rapid, specific and reliable assessment of the antitetanus immune status of horses and sheep. Compared with the indirect ELISA, the double antigen ELISA has the advantage of species-independent testing of sera. Thanks to its test design, it is more specific since the detected antibodies are forced to bind tetanus toxoid twice. In addition, it is very sensitive to tetanus antibodies, enabling the detection of low antibody titres, in range which is relevant for the assessment of the protective status (tetanus toxin neutralising antibodies). T...
Serum amyloid A isoforms in serum and synovial fluid in horses with lipopolysaccharide-induced arthritis.
Veterinary immunology and immunopathology    December 5, 2005   Volume 110, Issue 3-4 325-330 doi: 10.1016/j.vetimm.2005.10.012
Jacobsen S, Niewold TA, Halling-Thomsen M, Nanni S, Olsen E, Lindegaard C, Andersen PH.The aim of the study was to determine the intraarticular serum amyloid A (SAA) response pattern in horses with inflammatory arthritis. Inflammatory arthritis was induced by injection of lipopolysaccharide (LPS) into the radiocarpal joint of four horses. Serum and synovial fluid (SF) samples were collected before and at 4, 8, 12, 24, 48, 72, 96, and 144 h after injection. Concentrations of SAA were measured by immunoturbidometry, and expression of SAA isoforms was visualized by denaturing isoelectric focusing and Western blotting. The LPS injection caused systemic and local clinical signs of in...
Isolation of equine herpesvirus-2 from the lung of an aborted fetus. Galosi CM, de la Paz VC, Fernández LC, Martinez JP, Craig MI, Barrandeguy M, Etcheverrrigaray ME.This study describes the isolation of equine herpesvirus-2 (EHV-2) from the lung of an aborted equine fetus in Argentina. The isolated virus was confirmed as EHV-2 by indirect immunofluorescence using a rabbit anti-EHV-2 polyclonal antiserum and by virus-neutralization test using an equine polyclonal antibody against EHV-2. Restriction endonuclease DNA fingerprinting with BamHI also confirmed the identity of the virus as EHV-2. Furthermore, viral nucleic acid was detected by polymerase chain reaction from the original lung sample and from the DNA obtained from cells infected with the virus iso...
Black walnut extract-induced laminitis in horses is associated with heterogeneous dysfunction of the laminar microvasculature.
Equine veterinary journal    November 22, 2005   Volume 37, Issue 6 546-551 doi: 10.2746/042516405775314781
Peroni JF, Harrison WE, Moore JN, Graves JE, Lewis SJ, Krunkosky TM, Robertson TP.Equine laminitis purportedly involves haemodynamic dysfunction at the level of the laminar vasculature. However, to date, no studies have been performed characterising the function of laminar arteries and veins during the prodromal stages of equine laminitis. Objective: That the prodromal stages of laminitis are associated with contractile dysfunction of the equine laminar vasculature. Objective: To assess contractile function of laminar arteries and veins to phenylephrine (PE) and 5-hydroxytryptamine (5-HT). Methods: Horses were administered black walnut heartwood extract (BWHE) or water (con...
Cytogenetic mapping of immunity-related genes in the domestic horse.
Animal genetics    November 19, 2005   Volume 36, Issue 6 507-510 doi: 10.1111/j.1365-2052.2005.01348.x
Musilova P, Kubickova S, Vychodilova-Krenkova L, Kralik P, Matiasovic J, Hubertova D, Rubes J, Horin P.Chromosomal locations of 19 horse immunity-related loci (CASP1, CD14, EIF5A, FCER1A, IFNG, IL12A, IL12B, IL12RB2, IL1A, IL23A, IL4, IL6, MMP7, MS4A2, MYD88, NOS2A, PTGS2, TFRC and TLR2) were determined by fluorescence in situ hybridization. For IFNG and PTGS2, this study is a confirmation of their previously reported position. In addition, microsatellite (HMBr1) was localized in the same region as IFNG. All genes were assigned to regions of conserved synteny and the data obtained in this study enhance the comparative human-horse map. Cytogenetic localization of IL6 to ECA4q14-q21.1 suggested a...
cDNA sequence of the horse (Equus caballus) LAMA3 gene and characterization of two intronic SNP markers.
DNA sequence : the journal of DNA sequencing and mapping    November 17, 2005   Volume 16, Issue 6 468-473 doi: 10.1080/10425170500287674
Milenkovic D, Mata X, Chadi S, Guérin G.Laminins are large heterotrimeric basement membrane glycoproteins composed of alpha, beta and gamma chains. The Laminin 5 isoform has an alpha3beta3gamma2 composition and is essential for the adhesion of basal keratinocytes to the underlying epithelial basement membrane where it is mainly located. Mutations in the genes coding for the 3 chains have been associated with a severe skin blistering disease, Herlitz's junctional epidermolysis bullosa (JEB), observed in different species as man, dog, cat and horse. In this study, we report the sequence of the 5.2 kb horse laminin alpha 3 cDNA (LAMA3)...
Recovery of Swedish Equine arteritis viruses from semen by cell culture isolation and RNA transfection.
Journal of virological methods    November 16, 2005   Volume 133, Issue 1 48-52 doi: 10.1016/j.jviromet.2005.10.017
Mittelholzer C, Johansson I, Olsson AK, Ronéus M, Klingeborn B, Belák S.Recovery of infectious Equine arteritis virus (EAV) from the semen of persistently infected Swedish stallions was attempted by classical cell culture isolation and by transfection of extracted total RNA. Whereas virus from semen samples stored for several months at -20 degrees C or from extended semen could only be recovered by transfection of extracted RNA, isolation in cell culture was achieved readily with fresh, unextended semen stored at -70 degrees C or directly used after sampling. In parallel, the viruses were examined in the variable region of the large glycoprotein GP5 by nested RT-P...
Co-infection between influenza virus and flagellated bacteria.
Revista do Instituto de Medicina Tropical de Sao Paulo    November 16, 2005   Volume 47, Issue 5 275-280 doi: 10.1590/s0036-46652005000500007
Mancini DA, Mendonça RM, Dias AL, Mendonça RZ, Pinto JR.Trypsin is required in the hemagglutinin (HA) cleavage to in vitro influenza viruses activation. This HA cleavage is necessary for virus cell entry by receptor-mediated endocytosis. Bacteria in the respiratory tract are potential sources of proteases that could contribute to the cleavage of influenza virus in vivo. From 47 samples collected from horses, pigs, and from humans, influenza presence was confirmed in 13 and these samples demonstrated co-infection of influenza with flagellated bacteria, Stenotrophomonas maltophilia from the beginning of the experiments. Despite treatment with antibio...
Endocytosis and a low-pH step are required for productive entry of equine infectious anemia virus.
Journal of virology    November 12, 2005   Volume 79, Issue 23 14482-14488 doi: 10.1128/JVI.79.23.14482-14488.2005
Brindley MA, Maury W.Recently, it has become evident that entry of some retroviruses into host cells is dependent upon a vesicle-localized, low-pH step. The entry mechanism of equine infectious anemia virus (EIAV) has yet to be examined. Here, we demonstrate that wild-type strains of EIAV require a low-pH step for productive entry. Lysosomotropic agents that inhibit the acidification of internal vesicles inhibited productive entry of EIAV. The presence of ammonium chloride (30 mM), monensin (30 microM), or bafilomycin A (50 nM) in the medium dramatically decreased the number of EIAV antigen-positive cells. We foun...
The effect of enzyme treatment on the in vitro fermentation of lucerne incubated with equine faecal inocula.
The British journal of nutrition    November 10, 2005   Volume 94, Issue 5 771-782 doi: 10.1079/bjn20051561
Murray JA, Longland AC, Moore-Colyer MJ, Dunnett C.A series of experiments was conducted to determine the effects of a fibrolytic enzyme preparation (enzyme 1; E1) on the in vitro fermentation of lucerne incubated with equine faecal inocula. In experiment 1, high-temperature-dried (HT) lucerne was treated with five levels of E1 (0 to 2.4 ml/g DM) and incubated at 50 degrees C for 20 h. Samples then received a simulated foregut digestion (SFD) treatment before DM and NSP analysis. In experiment 2, HT lucerne was treated with the same enzyme levels used in experiment 1. Samples were then split into two groups; plus or minus an SFD treatment befo...
Effects of radial shock waves on membrane permeability and viability of chondrocytes and structure of articular cartilage in equine cartilage explants.
American journal of veterinary research    November 9, 2005   Volume 66, Issue 10 1757-1763 doi: 10.2460/ajvr.2005.66.1757
Byron CR, Benson BM, Stewart AA, Stewart MC.To investigate in vitro effects of radial shock waves on membrane permeability, viability, and structure of chondrocytes and articular cartilage. Methods: Cartilage explants obtained from the third metacarpal and metatarsal bones of 6 horses. Methods: Equine cartilage was subjected to radial shock waves and then maintained as explants in culture for 48 hours. Treatment groups consisted of a negative control group; application of 500, 2,000, and 4,000 impulses by use of a convex handpiece (group A); and application of 500, 2,000, and 4,000 impulses by use of a concave handpiece (group B). Effec...
Differences in tumor necrosis factor (TNF)alpha and TNF receptor-1-mediated intracellular signaling factors in normal, inflamed and scar-formed horse tendons.
The Journal of veterinary medical science    November 9, 2005   Volume 67, Issue 10 985-991 doi: 10.1292/jvms.67.985
Hosaka Y, Kirisawa R, Ueda H, Yamaguchi M, Takehana K.Tumor necrosis factor (TNF) receptors (TNF-R)-mediated cell survival or apoptosis has been demonstrated in many cells, but little is known about survival or apoptotic signals via TNF-R1 in tendinocytes. In this study, we focused on four signaling factors, TNFalpha, TNF-R1, TNFR-associated factor2 (TRAF2) and caspase-3, in order to elucidate the signaling events in tendinocytes. Samples were obtained from normal, inflamed and scar-formed equine superficial digital flexor tendons. To detect these signaling factors, samples were subjected to immunohistochemistry and Western blot analysis, and som...
Genomic characterization of MHC class I genes of the horse.
Immunogenetics    November 8, 2005   Volume 57, Issue 10 763-774 doi: 10.1007/s00251-005-0034-9
Tallmadge RL, Lear TL, Antczak DF.The availability of a contig of bacterial artificial chromosome (BAC) clones spanning the equine major histocompatibility complex (MHC) made possible a detailed analysis of horse MHC class I genes. Prior to this study, only a single horse MHC class I gene had been sequenced at the genomic level. Although many ( approximately 60) MHC class I cDNA sequences had been determined and published, from this information, it was not possible to determine how many class I loci are expressed in horses or to assign individual sequences to allelic series. In this study, 15 MHC class I genes were identified ...
Prepackaging of equine semen in goblets prior to cryopreservation.
Animal reproduction science    November 3, 2005   Volume 89, Issue 1-4 248-250 
Love CC, White RD, Varner DD.No abstract available
Evaluation of slow cooling after centrifugation and glycerol addition at 22 degrees C versus direct freezing of semen in stallions with good and poor sperm longevity.
Animal reproduction science    November 3, 2005   Volume 89, Issue 1-4 299-302 
Knop K, Hoffmann N, Rath D, Sieme H.No abstract available
Cryopreservation of stallion semen collected from good and poor freezers using a directional freezing device (Harmony CryoCare–Multi Thermal Gradient 516).
Animal reproduction science    November 3, 2005   Volume 89, Issue 1-4 291-294 
Zirkler H, Gerbes K, Klug E, Sieme H.No abstract available
Effect of extender and method of preservation on motility of cooled stallion spermatozoa.
Animal reproduction science    November 3, 2005   Volume 89, Issue 1-4 281-283 
Rota A, Magelli C, Impeduglia R, Panzani D, Camillo F.No abstract available
Effects of cushioned centrifugation technique on sperm recovery and sperm quality in stallions with good and poor semen freezability.
Animal reproduction science    November 3, 2005   Volume 89, Issue 1-4 294-297 
Knop K, Hoffmann N, Rath D, Sieme H.No abstract available
Effect of antibiotics on viability and fertility of equine semen cooled to 5 degrees C.
Animal reproduction science    November 3, 2005   Volume 89, Issue 1-4 277-280 
Macedo LP, Papa FO, Gomes GM, Melo CM, Oliveira JV, Dellaqua JA.No abstract available
Effect of storage and temperature on in vitro stallion sperm parameters and fertility rate.
Animal reproduction science    November 3, 2005   Volume 89, Issue 1-4 318-321 
Chanavat E, Vidament M, Defoin L, Duchamp G, Levillain N, Yvon JM, Le Vern Y, Kerboeuf D, Magistrini M.No abstract available
Relations between low rates of in vitro fertilization and induction of sperm acrosome reactions by zona pellucida in the equine species.
Animal reproduction science    November 3, 2005   Volume 89, Issue 1-4 270-271 
Mugnier S, Magistrini M, Sachet L, Lahuec C, Yvon JM, Meyers S, Goudet G.No abstract available
Effects of hyaluronic acid supplementation on motility of stallion spermatozoa after cryopreservation.
Animal reproduction science    November 3, 2005   Volume 89, Issue 1-4 284-285 
Mari G, Iacono E, Rizzato G, Merlo B, Belluzzi S.No abstract available
Improvement of the freezability of sex-sorted stallion spermatozoa.
Animal reproduction science    November 3, 2005   Volume 89, Issue 1-4 315-318 
Buss H, Clulow J, Sieme H, Maxwell WM, Morris LH, Sieg B, Struckmann C, Rath D.No abstract available
Improvement of stallion semen post-thaw motility with the association dimethyl formamide and methyl formamide as cryoprotectors.
Animal reproduction science    November 3, 2005   Volume 89, Issue 1-4 286-288 
Carmo MT, Papa FO, Medeiros AS, Araujo GH, Alvarenga MA.No abstract available
Evaluation of a cushioned centrifugation technique for processing equine semen for freezing.
Animal reproduction science    November 3, 2005   Volume 89, Issue 1-4 245-248 
Ecot P, Decuadro-Hansen G, Delhomme G, Vidament M.No abstract available