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.
Marr K, Marsh K, Hernandez L, Cunningham FM, Lees P.Leukotrienes, products of the 5-lipoxygenase pathway of arachidonic acid metabolism, possess properties consistent with their involvement in a range of inflammatory diseases. In this study the pharmacokinetics and pharmacodynamics of the selective 5-lipoxygenase inhibitor, fenleuton, have been examined in the horse. Orally administered fenleuton (four 5 mg kg(-1) doses, given once daily) was absorbed from the gastrointestinal tract, and penetrated readily into tissue cage exudate, the ratio of the plasma:exudate AUC0-48h being 0.90+/-0.02 (n=6). Ionophore-stimulated leukotriene (LT) B4 synthes...
Howard RD, McIlwraith CW, Trotter GW, Nyborg JK.To clone equine interleukin 1 alpha (IL-1 alpha) and equine interleukin 1 beta (IL-1 beta) and determine their full-length cDNA sequences. Methods: The mRNA isolated from lipopolysaccharide-stimulated cultured equine monocytes was reverse transcribed, and a cDNA library was constructed in a lambda phage. The cDNA library was screened by means of plaque hybridization with radiolabeled human IL-1 alpha and IL-1 beta cDNA probes. The cDNA nucleotide sequences for equine IL-1 alpha and equine IL-1 beta were determined by use of the dideoxy chain termination technique. The cDNA sequences were analy...
Barton MH, Bruce EH, Moore JN, Norton N, Anders B, Morris DD.To test efficacy of murine monoclonal, rabbit polyclonal recombinant equine or human tumor necrosis factor-alpha (rETNF or rHTNF, respectively) antibodies to inhibit native equine tumor necrosis factor (TNF) activity. Methods: 8 and 18 healthy adult horses for parts 1 and 2 of the study, respectively. Methods: In part 1, supernates from endotoxin-activated peritoneal macrophages were incubated with various dilutions of each rETNF antibody and subsequently tested for TNF activity. Serum was also obtained from a horse 1 hour after infusion with 20 ng of endotoxin/kg of body weight and was incuba...
Otte K, Choudhury D, Charalambous M, Engström W, Rozell B.The equine IGF2 gene has been cloned and characterised. It spans a 9 kb region, which is substantially less than the corresponding human gene. Three coding exons and three untranslated leader exons, all highly homologous to those in other species, were identified. Downstream of the polyadenylation site in exon 6, a dinucleotide repeat sequence was identified. Three putative promoters (P1-P3) were localised in the 5' region of the gene. RNase protection analysis revealed two active promoters in fetal tissues, P2 and P3, whereas P3 was the only promoter active in adult tissues. This represents a...
Willbold D, Metzger AU, Sticht H, Gallert KC, Voit R, Dank N, Bayer P, Krauss G, Goody RS, Rösch P.Lentiviral transactivator (Tat) proteins are essential for viral replication. Tat proteins of human immunodeficiency virus type 1 and bovine immunodeficiency virus form complexes with their respective RNA targets (Tat responsive element, TAR), and specific binding of the equine anemia virus (EIAV) Tat protein to a target TAR RNA is suggested by mutational analysis of the TAR RNA. Structural data on equine infectious anemia virus Tat protein reveal a helix-loop-helix-turn-helix limit structure very similar to homeobox domains that are known to bind specifically to DNA. Here we report results of...
Prieto D, Simonsen U, Hernández M, GarcÃa-Sacristán A.1. Penile small arteries (effective internal lumen diameter of 300 600 microm) were isolated from the horse corpus cavernosum and mounted in microvascular myographs in order to investigate the mechanisms underlying the endothelium-dependent relaxations to acetylcholine (ACh) and bradykinin (BK). 2. In arteries preconstricted with the thromboxane analogue U46619 (3-30 nM), ACh and BK elicited concentration-dependent relaxations, pD2 and maximal responses being 7.71+/-0.09 and 91+/-1 % (n=23), and 8.80+/-0.07 and 89+/-2% (n=24) for ACh and BK, respectively. These relaxations were abolished by me...
Breidenbach A, Schlumbohm C, Harmeyer J.The aim of the present study was to investigate the importance of putative regulatory factors of the calcium (Ca) and inorganic phosphate (P(i)) homeostatic system in the horse. The concentrations of Ca, P(i), vitamin D metabolites, parathyroid hormone (PTH), the activity of the alkaline phosphatase (AP) and the concentration and binding properties of vitamin D binding protein (DBP) were measured in the plasma. In addition, the ability of the renal cortex to hydroxylate calcidiol into 24,25(OH)2D3 and 1,25(OH)2D3 was evaluated in vitro. The plasma concentration of Ca (3.2 +/- 0.15 mmol.L-1, N ...
Hinrichs K.When recovered from the follicle, horse oocytes may be categorised as having either a compact or an expanded cumulus. Cumulus expansion is strongly associated with follicle atresia. Oocytes with expanded and compact cumuli have similar proportions in the germinal vesicle stage when recovered from the follicle. However, during in vitro culture, a higher proportion of oocytes with expanded cumuli mature, and they do so more quickly, than do oocytes with compact cumuli. Using Hoechst 33258 to label chromatin, in the germinal-vesicle stage horse oocytes can be divided into those in which the nucle...
Kato H, Seidel GE, Squires EL, Wilson JM.In vitro matured horse oocytes with a first polar body (n = 68) were each injected with a single spermatozoon and divided into 2 groups: Group 1 oocytes were treated with 10 microM calcium ionophore A23187 for 5 min while Group 2 oocytes received no activation treatment. After culture in vitro for 2 days, significantly more oocytes treated with A23187 (5/24, 21%) cleaved than oocytes without activation treatment (2/44, 5%, P<0.05). All 7 cleaved zygotes from both treatment groups were transferred to recipient mares but no pregnancies resulted.
Huhtinen M, Lagneaux D, Koskinen E, Palmer E.Seventy-five embryos were collected 6 days after ovulation. Sixty embryos were frozen in straws using glycerol as the cryoprotectant in an automatic freezer. In Experiment 1 the freezing and thawing media were supplemented with 1.3 g/l PVP; in Experiment 2 the supplement was 5% FCS. The embryos were thawed for 30 s at +37 degrees C in a waterbath. In Experiment 1 glycerol was removed from 10 embryos in 6 steps. In 10 other embryos, glycerol and sucrose were both removed from the medium in 6 steps. After glycerol and sucrose removal, the embryos were stained with 4',6'-diamidino-2-phenylindole ...
Young CA, Squires EL, Seidel GE, Kato H, McCue PM.Larger grade 1 or 2 (1 = excellent,.... 4 = degenerate) equine embryos that ranged in diameter from 300 to 680 microm and were recovered from mares on Day 7 or 8 after ovulation, were randomly assigned to 3 widely divergent cryopreservation treatments. Treatment 1 consisted of cooling from -6 degrees C to -35 degrees C at 0.5 degrees C per min followed by plunging into liquid nitrogen, with a one-step addition and a 4-step removal of 1.0 M glycerol. Treatment 2 (step-down equilibration) consisted of a 2-step addition of glycerol to 4.0 M followed by a decrease to 2.0 M prior to freezing, with ...
Ferreira JC, Meira C, Papa FO, Landin e Alvarenga FC, Alvarenga MA, Buratini J.Six or 7-day-old equine embryos were divided into 4 groups; Group 1, n = 15, Day 7 embryos destined for immediate transfer; Group 2, n = 15, Day 6 embryos destined for deep-freezing with glycerol plus sucrose as cryoprotectant; Group 3, n = 10, Day 6 embryos destined for deep-freezing with glycerol plus 1,2-propanediol as cryoprotectant and Group 4, n = 3, fresh embryos destined for ultrastructural analysis. All the frozen/thawed embryos were transferred to recipient mares, except 3 embryos in Group 3 that were subjected to ultrastructural analysis. After thawing the cryoprotectants were remov...
Alm H, Torner H, Kanitz W, Becker F, Hinrichs K.The object of this study was to compare 4 different methods of oocyte recovery from mares; 1) transvaginal follicle aspiration in vivo; 2) follicle aspiration in vitro; 3) oocyte recovery by isolation of follicles in vitro and 4) follicle scraping in vitro. Oocyte recovery was highest after follicle scraping (71.1%) and follicle isolation and rupture (61.3%). Follicle aspiration in vitro and in vivo yielded oocytes on 31.2% and 19.3% of occasions, respectively. The output of different types of cumulus-oocyte-complexes was different among the methods; the portion of compact cumulus-oocyte-compl...
Aguilar JJ, Woods GL, Miragaya MH, Olsen LM.The object of this experiment was to estimate the number and type of living cells in oviductal masses of mares. Oviducts of abattoir mares were dissected, divided into 3 sections, and flushed individually. Oviductal masses were recovered from 220 of 250 mares and from 389 of 500 oviducts. A greater number of masses was recovered from the left than the right oviducts. A higher percentage of masses was recovered from the ampullary-isthmic junction than from the ampulla or isthmus. The number of masses increased slightly with increasing mare age and was weakly correlated with the number of unfert...
Mlodawska W, Okólski A.From the ovaries of 256 slaughtered mares a total of 1713 follicles were isolated from which 1641 (95.8%) oocytes were recovered (6.4/mare). A total of 564 follicles and oocytes were evaluated for the degree of vascularisation of the follicle wall, the appearance of the follicular fluid and the location and morphology of the cumulus-oocyte-complex. Follicles with a diameter of >10 mm displayed more numerous, well branched and more pronounced blood vessels than the smaller ones (4-10 mm diameter) and most of them contained clear, yellowish fluid with few granulosa cells. The percentage of oo...
Lagneaux D, Huhtinen M, Koskinen E, Palmer E.Equine embryos recovered on Day 6 after ovulation were cooled to +4 degrees C, or frozen with AFP alone or together with glycerol. Twenty embryos (140-200 microm in diameter) were randomly assigned to 6 treatment groups. In the first 3 groups, the embryos were cooled from room temperature to +4 degrees C at a rate of 3 degrees C/min and warmed again at a rate of 32 degrees C/min in a programmable freezer. In the second 3 groups, the embryos were frozen using a standard protocol, stored in liquid nitrogen for 5-7 days and then thawed in a 37 degrees C waterbath. After cooling/warming or freezin...
Meintjes M, Graff KJ, Paccamonti D, Eilts BE, Paul JB, Thompson DL, Kearney MT, Godke RA.When aspirating ovarian follicles in pregnant mares to obtain oocytes for in vitro fertilisation (IVF), the effect of the manipulation on circulating concentrations of progesterone may be an important consideration in terms of the maintenance of pregnancy. The object of this study was to compare the effects of 3 different forms of transvaginal ultrasound-guided follicle aspiration (Treatment 1, no aspiration, n = 4; Treatment 2, aspirate only follicles > or =20 mm in diameter, n = 7; Treatment 3, aspirate all visible follicles, n = 7) on peripheral plasma progesterone concentrations between Da...
Liang FT, Granstrom DE, Zhao XM, Timoney JF.Sarcocystis neurona is the etiologic agent of equine protozoal myeloencephalitis (EPM). Based on an analysis of 25,000 equine serum and cerebrospinal fluid (CSF) samples, including samples from horses with neurologic signs typical of EPM or with histologically or parasitologically confirmed EPM, four major immunoblot band patterns have been identified. Twenty-three serum and CSF samples representing each of the four immunoblot patterns were selected from 220 samples from horses with neurologic signs resembling EPM and examined for inhibitory effects on the infectivity of S. neurona by an in vi...
Smith TA, Davis E, Carpenter S.Lentiviruses replicate in cells of the immune system, and activation of immune cells has been shown to modulate virus replication. To determine the effects of macrophage activation on replication of equine infectious anaemia virus (EIAV), primary horse macrophage cultures (HMCs) were established from 20 different horses, infected with an avirulent strain of EIAV, and stimulated with 5 microg/ml of bacterial endotoxin. Supernatants collected from HMCs were assayed for the presence of tumour necrosis factor (TNF-alpha) and for production of infectious virus. Results indicated that EIAV replicati...
Nichani AK, Sharma RD, Sarup S.Efficacy of medium RPMI-1640 (supplied by Gibco USA, Centron and Hi-media) supplemented with horse, donkey, sheep and goat sera was evaluated for in vitro propagation of Theileria annulata (Hisar) infected bovine mononuclear cells. The results were compared with the growth rate in RPMI-1640 supplemented with foetal bovine serum (Gibco). RPMI-1640 (Gibco) proved to be the best medium for in vitro cultivation of the parasite infected cells. Foetal bovine serum could be easily, safely and reliably substituted with goat and sheep sera in the growth medium. Horse and donkey sera also gave comparabl...
Sternberg S.Swab samples were collected from the oral cavity of 174 horses in 10 farms and cultured selectively for Actinobacillus equuli. A. equuli could be isolated from 37% of all samples, varying between 12 and 88% in the different farms. Eight horses were sampled repeatedly for several days, with a variation in isolation frequency between 50 and 88%. Isolates were compared by restriction enzyme digestion and Pulsed-Field Gel Electrophoresis. A high degree of strain variability was found within each horse population as well as some variability over time between strains isolated from the same horse.
Hare JE, Viel L, Conlon PD, Marshall JS.Pulmonary mast cells (PMC) are important components of the inflammatory process in equine allergic lung diseases such as heaves. Very little, however, is known of the degranulation kinetics of these cells and thus, their pathophysiologic role remains largely speculative. The purpose of this study was to develop a repeatable protocol for in vitro equine PMC degranulation. Five mature horses (sex: 2 M, 3 F; age: 8.8 +/- 6.5 y), historically free of pulmonary disease and normal on clinical respiratory examination, arterial blood gas analysis, pulmonary mechanics testing and histamine inhalation c...
Hinrichs K, Matthews GL, Freeman DA, Torello EM.Oocytes were collected from dominant preovulatory follicles of donor mares 24 hours after administration of human chorionic gonadotropin. Oocytes were incubated in vitro for 12 or 18 hours before transfer to recipient mares, representing maturation times after human chorionic gonadotropin administration of 36 and 42 hours, respectively. Pregnancy rates after transfer were 4 of 5 in the 36-hour group and 2 of 3 in the 42-hour group. The overall pregnancy rate achieved (6/8 mares) indicated that oocyte transfer may be useful clinically.
Bailey SR, Elliott J.Cumulative concentration response curves to 5-hydroxytryptamine (5-HT; 10(-10)-10(-4) mol/l) were constructed using isolated rings of equine digital, facial, tail and coronary arteries (endothelium intact). 5-HT was 17.7 and 41 times more potent as a vasoconstrictor of digital arteries than facial and tail arteries respectively. Removal of the endothelium increased the vasoconstrictor potency of 5-HT in the facial artery by 3.7-fold (P<0.05) but did not alter the sensitivity of digital arteries to 5-HT. Coronary arteries failed to contract to 5-HT. Coronary arteries pre-contracted with U440...
Graham JK.Methodologies to capacitate bovine spermatozoa, induce the acrosome reaction, and fertilize bovine oocytes in vitro have been established. The capability to do the same with stallion spermatozoa, however, is not available. Several different methods have been used to capacitate stallion spermatozoa with variable results. More basic research needs to be done to establish in vitro conditions necessary to capacitate and induce an acrosome reaction in stallion spermatozoa. Although much progress can be expected in this area, it is unlikely that the general practitioner will use these technologies i...
Bertrand ML, Harris DC.To test whether the reactivity of ferritin iron is affected by the heat treatment used in ferritin isolation, we prepared ferritin from the same horse spleen with or without heating. Both samples exhibited similar reactivity upon reduction or chelation of iron.
Kujawa TJ, van Doorn DA, Wambacq WA, Hesta M, Pellikaan WF.The in vitro gas production technique (IVGPT) has been a valuable tool in ruminant nutrition research for decades and has more recently been used in horse nutrition studies to investigate fermentation activities of the equine hindgut though primarily using feces as inoculum. This study was conducted to evaluate the use of equine rectal content in the IVGPT system as a viable inoculum that can be considered representative of the activities throughout the equine hindgut. Additionally, the study was conducted to measure the effects on fermentation kinetics and end-product production using inoculu...
Pycock JF, Allen WE, Porter DJ, Boyd EH.Two independent assay systems were used to study the effect of three antibacterial preparations on in vitro morphology and chemotaxis of equine neutrophils. Incubation of neutrophils with high (200 micrograms/ml) and medium (20 micrograms/ml) concentrations of neomycin impaired their response to standard chemoattractants. Trimethoprim/sulfadoxine (0.4/2.0 micrograms/ml-40/200 micrograms/ml) and benzylpenicillin (0.25-25 micrograms/ml) had no effect. Neutrophils collected from geldings 2 and 24 h after neomycin (5 mg/kg) administration had impaired responses to standard chemoattractants. Benzyl...
Akridge HD, Rankin SC, Griffeth GC, Boston RC, Callori NE, Morris DO.Meticillin-resistant Staphylococcus aureus (MRSA) strain USA 500 predominately colonizes horses and people working with them. Previous studies demonstrate that some Staphylococcus species exhibit higher affinity for corneocytes of specific mammalian species. Objective: The objective was to determine the relative affinities of various MRSA strains, meticillin-susceptible S. aureus (MSSA) strains and a meticillin-susceptible Staphylococcus pseudintermedius (MSSP) for equine corneocytes. We hypothesized that MRSA strain USA 500 would exhibit greater adhesion than other staphylococcal strains te...
INGRAM DG.A method is described for the quantitative measurement of the reactions between sensitized cells and horse complement and between alexinated cells and conglutinin. The method is laborious but its application has allowed the determination of the optimal times of the reactions at various temperatures. The results obtained in these experiments indicate that the alexinated configuration with which conglutinin and immuno-conglutinin react is not one of the recognized intermediates formed during the process of immune haemolysis.
Diethelm AG, Chambers LM, Balch CM, Phillips SJ.Anti-human thymocyte cell membrane antibody prepared by hyperimmunization of the horse produced an antiserum capable of prolonging skin allografts in the rhesus monkey for an average of 26 days. Lymphocyte depletion was present in paracortical areas of mesenteric lymph nodes of these animals after 28 days of treatment; the intravenous administration was tolerated without ill effects. Immunofluorescent studies identified both broad specificity antibodies reacting with numerous human cell types as well as thymus-dependent (T) cell antibodies reactive with human thymocytes and peripheral T-cells....
Erice I, Gil L, Josa A, Echegaray A, Martinez F, Espinosa E.Mares (n = 39) were classified according to age as young (less than 1.5 yr, n = 17) or old (more than 1.5 yr, n = 22) and sacrificed. Ovaries were measured and weighed, and the number of follicles and CL were counted. Follicle size and distribution were recorded (external: > 20 mm, 5 mm, < 5 mm). External follicles were aspirated while internal follicles were sliced. The number and Type of oocytes recovered using each method were recorded. Oocyte recovery rates (oocytes/ovary) resulted in a mean of 0.92 oocytes by aspiration and 1.36 oocytes by additional slicing. The mean numbers of av...
GarcÃa-Sacristán A, Casanueva CR, Castilla C, Labadia A.The presence and types of alpha and beta adrenergic receptors in the urethra and prostate of the horse were studied in vitro using adrenergic agonist and antagonist drugs. The existence of these receptors was shown. This finding was based on the observation that the contractile action was mediated by adrenergic receptors of alpha-1 type, although in the prostate alpha-2 type receptors also participated. Relaxation in both tissues was controlled by receptors of the beta-2 type.
Kingston JK, Bayly WM, Meyers KM, Sellon DC, Wardrop KJ.There is evidence that equine platelet reactivity is altered by strenuous exercise. Changes in platelet reactivity could impact haemostasis following exercise-induced injury and may play a role in the pathophysiology of exercise-induced pulmonary haemorrhage. Interpretation of results of previous studies is hindered by potential in vitro-induced changes in platelet activity through the choice of anticoagulant and the use of platelet inhibitors. The present study was undertaken to re-evaluate the effect of exercise on equine platelets using methodologies that minimise in vitro-induced changes i...
Pycock JF, Allen WE.Streptococcal endometritis was induced experimentally in pony mares during oestrus. Uterine fluid was collected 30, 60, 120 or 240 minutes later and tested for its effect on the in vitro morphology and chemotaxis of equine neutrophils by two independent methods. The maximal response occurred between 30 and 60 minutes after infection and persisted until 240 minutes. The chemo-attractant contained both heat labile and heat stable components and the latter appeared to be active at low concentrations.
Santos MP, Gutierrez-Nibeyro SD, Horn GP, Hicke JD, Stewart MC, Schaeffer DJ.To compare the mechanical properties of laryngeal tie-forward (LTF) constructs prepared with different suture materials and suture placement patterns during single load to failure testing. Methods: Larynges harvested from 50 horse cadavers and 5 intact horse cadavers. Methods: In vitro LTF constructs were created by a standard technique with polyester sutures, a standard technique with polyethylene sutures, a modified technique with metallic implants and polyester sutures, a modified technique with metallic implants and polyethylene sutures, or a modified tie-off technique with polyester sutur...
Takeuchi K, Shibata N, Senda N.Myosin was isolated from leucocytes in horse arterial blood by the same procedures used for the isolation of myosin from skeletal muscle. The Ca2+-, EDTA-, and Mg2+-ATPase [EC 3.6.1.3] activities of the protein was 0.148, 0.147, and 0.001 mumoles/min/mg, respectively, in 0.5 M KCl at pH 7.0 and 25 degrees. The Ca2+-ATPase activity decreased with decrease in the ionic strength. No difference was found between leucocyte myosin and skeletal myosin in the pH profiles of Ca2+- and EDTA-ATPases. The rate and amount of the initial burst of Pi liberation of leucocyte myosin were 0.002 mumoles/min/mg a...
Quirino M, Jakop U, Mellagi APG, Bortolozzo FP, Jung M, Schulze M.For a more practically applicable analysis of different sperm characteristics, this study aimed to develop a 5-color flow cytometry (FC) panel to concurrently analyze four sperm parameters in liquid boar and stallion semen, using also a DNA-marker for selecting sperm cell events. From each of thirty extended boar semen doses and twelve stallion semen doses, six aliquots were taken. For evaluating mitochondrial activity (A), degree of lipid disorder of plasma membrane (B), integrity of plasma membrane (C), acrosomal status (D) and marking DNA (E), five aliquots were individually stained with Rh...
Vörös K, Holmes JR, Gibbs C.This study was performed on 15 horses to validate the cardiac anatomy as imaged with two dimensional echocardiography (2DE) and to determine the accuracy of intracardiac measurements. 2DE images were recorded in five horses in life with a Diasonics DRF100 ultrasound system and a 3.5 MHz transducer in different tomographic planes. After slaughter, the hearts were fixed in 10 per cent formalin. To compare in vivo and in vitro results, the specimens were suspended in a water-filled tank and 2DE images were made using the same transducer positions as in life. In vitro 2DE and autopsy measurements ...
Freeman DE, Donawick WJ.Mucosa obtained from the cecum of healthy horses and incubated in vitro with 0.1 mM cycloleucine could accumulate this amino acid against an apparent concentration gradient after 60 and 120 minutes. Accumulation by the serosal (antiluminal) surface of the tissue was 3 times greater than accumulation by the mucosal (luminal) surface after 120 minutes (P less than 0.001). Cycloleucine accumulation was significantly reduced by Na deprivation after 60 minutes (P less than 0.05) and 120 minutes (P less than 0.01) and by anoxic conditions after 120 minutes (P less than 0.05). Transmucosal flux from ...
Rijkenhuizen AB, Sommerauer S, Fasching M, Velde K, Peham C.There are no studies on the ideal ligature technique for the spermatic cord. Objective: To compare the maximal resistance pressure in the testicular artery and the maximal tensile forces to produce failure of 2 different ligature techniques used for ligation of the equine spermatic cord. Methods: The capabilities of 2 types of ligatures, single knot loop and double knot loop, were assessed using a pressure-resistance test in testicular arteries and with an in vitro mechanical evaluation of the tensile strength by single cycle-to-failure testing. Results: In the pressure-resistance test, the me...
Aggarwal N, Holmes MA.The aim of this study was to characterise CD4+T-cells in equines, as these cells are pivotal in establishing immune responses or regulating established ones. Peripheral blood mononuclear cells from a pony immunised with ovalbumin were cultured in vitro in the presence of the specific antigen and autologous antigen presenting cells. During the antigen starvation phase, cells were maintained on recombinant equine IL-2. After 35 days of culture, most of the cells were CD4+, CD8-and sIg-. Cells proliferated specifically in the presence of antigen, as tested on day 42 of culture. These cells were a...
Frost AB, Larsen F, Ostergaard J, Larsen SW, Lindegaard C, Hansen HR, Larsen C.Development of suitable in vitro release models for formulation development as well as quality control purposes has to be initiated in the early design phase of injectable depots. Optimally, construction of an in vitro release model may lead to the establishment of in vitro in vivo correlations. By using a model compound (sodium diatrizoate, DTZ), the purpose of this study was to investigate the possibility of establishing in vitro in vivo relations between the DTZ disappearance profile obtained from the donor compartment of the rotating dialysis cell model and the joint disappearance profile ...
Gargiulo AM, Pedini V, Ceccarelli P, Lorvik S.In the present work, gustatory glands (von Ebner's glands) of the horse tongue were examined by means of five peroxidase-conjugated lectins (PNA, DBA, SBA, UEA I, WGA), with and without prior sialidase digestion, in order to investigate the presence and distribution of carbohydrate residues in secretory cells and duct cells. The most intense staining of secretory cells was observed with PNA after pre-treatment with neuraminidase. This indicates that the terminal trisaccharide sequence sialic acid- (alpha 2-->3, 6) galactosyl (beta 1-->3) N-acetylgalactosamine is the most frequent oligosa...
Wang L, Tong G, Liu H, Yang Z, Qiu H, Kong X, Wang M.Proviral DNA was extracted from donkey leukocyte infected with Chinese donkey leukocyte attenuated equine infectious anemia virus (DLA-EIAV), and peripheral blood lymphocytes (PBL) from a horse infected with the virulent EIAV strain Liaoning (EIAV L). The entire proviral DNA from both viruses was cloned and sequenced. The lengths of complete genomic sequences of DLA-EIAV and EIAV L provirus were 8266 bp and 8235 bp, respectively. Sequence comparison indicated that DLA-EIAV shares 97.0% and 97.5% in sequence homology with EIAV L and donkey-adapted EIAV (DA-EIAV), respectively. Lots of variation...
Kuwano A, Niwa H, Arai K.To establish a new system to isolate keratolytic bacteria from the hoof wall cavity () of a racehorse, we invented the horn powder agar-translucency (HoPAT) test and horn zymography (HZ). Using routine bacteriological techniques and these methods, we isolated five strains of keratolytic soil bacteria, which were then identified by means of 16S ribosomal RNA (rRNA) gene sequencing analysis. The findings from the study on the horse suggested that Brevibacterium luteolum played the main role in the local fragility of the hoof, eventually forming a in coordination with four other strains of kerat...
Grünig G, Winder C, Hulliger C, Witschi U, Hermann M, Jungi TW, von Fellenberg R.Increases in procoagulant activities (PCA) in equine lung macrophages were induced by non-adherent blood lymphocytes which were prestimulated with phytohaemagglutinin for 48 to 72 hours or by supernatants harvested from prestimulated blood lymphocyte cultures. However, prestimulated lymphocyte suspensions themselves expressed PCA which was most probably derived from contaminating monocytes. Because non-adherent cells from lymphocyte suspensions may have attached to adherent macrophages, cells within lymphocyte suspensions might have contributed to the PCAs expressed by lymphocyte-stimulated lu...
Noll CV, Kilcoyne I, Nieto JE, Thio T, Byrne BA.To determine the ability of 0.2% polyhexamethylene biguanide (PHMB)-impregnated gauze to inhibit the growth of bacteria isolated from equine infected sites. Methods: In vitro study. Methods: Nine bacterial isolates were obtained from cultures submitted from equine patients presenting with penetrating injuries of the hoof (n = 4), septic osteitis (n = 1), synovial sepsis (n = 1), wounds (n = 2), and incisional infection following laparotomy (n = 1). Two standardized strains were also included. A standard inoculum of each isolate was placed on 12 Muller-Hinton agar plates. Squares (2.5 cm ×â€...
Wallace KD, Selcer BA, Tyler DE, Brown J.Ninety-one equine aortic and cranial mesenteric arterial segments were evaluated ultrasonographically in a water bath. On the basis of pathologic evidence of verminous arteritis, arterial segments were classified into 4 categories, and the ultrasonographic characteristics of each group were evaluated. Normal arteries (class 1) were ultrasonographically characterized by a smooth luminal surface layer and uniform wall thickness and echogenicity. Arteries with only histopathologic evidence of verminous arteritis (class 2) were ultrasonographically characterized by a smooth luminal surface layer, ...
May SA, Hooke RE, Lees P.Although an E-series prostaglandin has previously been identified in equine inflammatory exudate, the identity of this eicosanoid as PGE2 has not been confirmed. The objective of this study was the specific characterisation of the prostaglandin produced by equine cells in the presence of an inflammatory stimulus. By using two radioimmunoassays, a relatively non-specific PGE2 assay and a more specific PGE1 assay, it has been possible to identify the E-series prostaglandin produced by equine chondrocytes and synovial cells, in response to a variety of stimuli, as PGE2.
Coignoul FL, Bertram TA, Roth JA, Cheville NF.Neutrophils from 4 pony foals, 3 lactating pony mares, and 3 nonlactating mares were evaluated ultrastructurally and by in vitro function tests. Neutrophils from foals had significantly (P = 0.05) less random migration than neutrophils from mares; values in tests for iodination and Staphylococcus aureus ingestion were also lower with foal neutrophils. Neutrophils from lactating mares had lower responses to iodination, antibody-dependent cell-mediated cytotoxicity, and random migration tests than did neutrophils from nonlactating mares. Ultrastructurally, granule concentration did not differ si...
Benanchi PL, Gazzei G, Giannozzi A.A simple two-step procedure for purifying F(ab)2 fragments of horse immunoglobulins is described. In the first step, the horse plasma is diluted, made up to 12% (w/v) with ammonium sulphate and digested with pepsin. In the second step, the previously dialyzed solution is chromatographed. Instead of a normal ion-exchange resin, a DEAE-cellulose, covalently linked to a synthetic vinyl polymer, was used (DEAE-Zeta-Prep). With this assembly it is possible to perform chromatography at a high flow-rate without the problems related to the use of large columns. The yield and purity of the final produc...
Wauters J, Franck T, Pille F, Martens A, Demeyere K, Sys S, Serteyn D, Gasthuys F, Meyer E.Myeloperoxidase (MPO) is a protein of interest due to its involvement in equine pathologies. Until now, results in equine diagnostic research were achieved through extracellular MPO detection. However, studying the cellular MPO content in neutrophils has revealed important insights in human diseases. This study aimed to develop a technique for the specific detection of MPO on the single cell level defining a flow cytometric protocol for the detection of both equine surface-bound and cellular MPO. Both indirect and direct labeling techniques are described which include the comparison of two sec...
