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Micropreparative high resolution purification of proteins by a combination of sodium dodecyl sulfate-polyacrylamide gel electrophoresis, isoelectric focusing, and membrane blotting. We report a simple, economical, and efficient protocol for protein purification from cells. First, proteins of cell lysates were separated by standard sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and electroblotted to protein-blotting membrane. The blots were stained with Coomassie blue or developed by immunoblotting to visualize specific proteins. The bands corresponding to those visible by immunoblotting were excised from the dye-stained blots and subjected to isoelectric focusing. The focused gel was stained with Coomassie blue. Finally, the stained bands were excise...
Assessment of viability and mitochondrial function of equine spermatozoa using double staining and flow cytometry. An objective double-staining method was developed to evaluate viability and mitochondrial function of stallion spermatozoa using flow cytometry. Sperm viability was assessed by propidium iodide (PI) exclusion, and mitochondrial function was measured by the intensity of rhodamine 123 (R123) fluorescence. Flow cytometry estimates of sperm viability measured by PI were equivalent (P > 0.05) to estimates made using Hoechst 33258 stain and fluorescent microscopy (% dead: 25 +/- 2.4 vs 21.5 +/- 3.5). The use of both PI and R123 was validated by addition of various proportions of freeze-shocked (m...
Evaluation of itraconazole-dimethyl sulfoxide ointment for treatment of keratomycosis in nine horses. To evaluate the efficacy of itraconazole-dimethyl sulfoxide ointment for treatment of keratomycosis in horses in the northeastern United States. Methods: Prospective clinical trial. Methods: 9 horses (10 affected eyes). Methods: All horses treated for keratomycosis at Cornell University between July 1994 and July 1996 were included in the study. The diagnosis of keratomycosis was confirmed by cytologic examination, and all horses were treated with 0.25 ml of a 1% itraconazole-30% dimethyl sulfoxide petrolatum-based ointment, applied to the affected eye every 4 hours. Results: Topical applicati...
Equine botulism. Botulism is a severe and often fatal disease in equine patients worldwide. Clostridium botulinum is a ubiquitous soil organism which produces a potent neurotoxin resulting in neuromuscular blockade and flaccid paralysis in affected animals. Definitive diagnosis is often impractical or impossible, leading to diagnosis and treatment based on clinical experience and presenting signs. Delay in case identification and treatment results in rapid deterioration of the patient. Treatment of recumbent equine botulism cases presents challenges due to patient size and requires intensive nursing care. This...
Surgical treatment of a comminuted articular fracture of the accessory carpal bone in a thoroughbred horse. The clinical, radiographic and ultrasonographic findings in a case of a comminuted articular fracture of the accessory carpal bone of a thoroughbred chaser are described, and its surgical treatment and aftercare are detailed. The horse made an uneventful recovery and successfully returned to racing.
Ageing draft and trotter horses by their dentition. The accuracy of ageing horses by their dentition was assessed by comparing the dental features with the known dates of birth of 212 trotter horses and 189 Belgian draft horses. The horses ranged in age from two to 25 years. In both breeds it was observed that the shedding of the incisors and the appearance of the dental stars were the most reliable features for age determination. In young animals, the dental configuration was similar in both breeds. With increasing age the incisor teeth of draft horses were more liable to abrasion than those of trotter horses. The sequential changes in appeara...
Immunogenicity and efficacy of baculovirus-expressed and DNA-based equine influenza virus hemagglutinin vaccines in mice. Two fundamentally different approaches to vaccination of BALB/c mice with the hemagglutinin (HA) of A/Equine/Kentucky/1/81 (H3N8) (Eq/KY) were evaluated, that is, administration of HA protein vs administration of HA-encoding DNA. Each vaccine was tested for its immunogenicity and ability to provide protection from homologous virus challenge. HA protein was synthesized in vitro by infection of Sf21 insect cells with a recombinant baculovirus. Intranasal administration of this vaccine induced virus-specific antibodies, as measured by enzyme-linked immunosorbent assay (ELISA), but did not induce ...
Development and validation of a series of three-dimensional finite element models of the equine metacarpus. Three-dimensional finite element (FE) models of the left metacarpi of five adult horses were developed from quantitative computed tomography data, using the algorithms of Keyak et al. (1990, J Biomed. Engng 12, 389-397). The metacarpi were then equipped with 12 rosette strain gauges and loaded non-destructively in a mechanical testing machine. The bones and the models were loaded in axial compression, with the load evenly distributed across the distal row of carpal bones, and with a point load placed mediad to the sagittal midline, to a load equivalent to three times body weight (-15 kN); and ...
An empirical study of the impact of complexity on participation in horserace betting. The aim of the research reported in this paper was to explore empirically whether levels of participation in horserace betting are affected by the complexity of the betting task. The study employed a systematic random sample of 1161 betting decisions made in UK offcourse betting offices during 1987. The research was conducted in a naturalistic setting where it was possible to grade complexity and to measure levels of participation. Complexity was defined in terms of both the number of alternatives in the decision-maker's choice set (number of horses in a race) and the complexity of the attribu...
The ICP0 protein of equine herpesvirus 1 is an early protein that independently transactivates expression of all classes of viral promoters. To assess the role of the equine herpesvirus type 1 (EHV-1) ICP0 protein (EICP0) in gene regulation, a variety of molecular studies on the EICP0 gene and gene products of both the attenuated cell culture-adapted Kentucky A (KyA) strain and the Ab4p strain were conducted. These investigations revealed that (i) the ICP0 open reading frame (ORF) of the KyA virus strain is 1,257 bp in size and would encode a protein of 419 amino acids, and in comparison to the ICP0 gene (ORF63) of the Ab4p strain of 1,596 bp (E. A. Telford, M. S. Watson, K. McBride, and A. J. Davison, Virology 189:304-316, 1992), ...
Anesthetic potency of desflurane in the horse: determination of the minimum alveolar concentration. To determine the minimum alveolar concentration (MAC) of desflurane (DES) in the horse. Methods: Prospective study. Methods: Six healthy adult horses (three males and three females) weighing 370 +/- 16 kg and aged 9 +/- 2 years old. Methods: Anesthesia was induced with DES vaporized in oxygen via a face mask connected to a large-animal, semiclosed anesthetic circle system. The horses were endotracheally intubated and positioned in right lateral recumbency. Inspired and end-tidal DES were monitored using a calibrated Ohmeda RGM 5250 multigas analyzer (Ohmeda-BOC, Spain). The MAC of desflurane t...
Carbonic anhydrase activity in different placenta types: a comparative study of pig, horse, cow, mink, rat, and human. The placenta has multiple functions, being the organ which provides oxygen and nutrients to the developing conceptus. In the placenta, the enzyme carbonic anhydrase (CA) may provide ions for exchange with Na+, K+, and Cl- in transepithelial movement of ions and fluid, as well as facilitating carbon dioxide diffusion. It can also be active in intermediary metabolism, such as gluconeogenesis, urea, and fatty acid synthesis. Placental material from pig, horse, cow, mink, rat, and human was therefore investigated, representing placenta types with variations in shape, internal architecture, and nat...
Incomplete nasomaxillary dysplasia in a foal. Atresia of the nasal punctum is the most common congenital anomaly for the equine nasolacrimal system. Nasomaxillary dysplasia has not been previously documented in foals, is of unknown etiology, and appears to be a rare condition. Conjunctivomaxillary sinostomy was successful in resolving the epiphora.
Clinical evaluation of the serodiagnostic value of enzyme-linked immunosorbent assay for Rhodococcus equi infection in foals. An enzyme-linked immunosorbent assay (ELISA) for detection of serum IgG antibodies against Tween 20-extracted antigen of strain ATCC 6939 was applied in Hidaka, Japan to a total of 752 sick foals showing a variety of signs of infectious disease. An optical density (OD) value of more than 0.3 was tentatively fixed to be positive on the basis of readings made of healthy horse sera in previous studies. During a 2 year study, 138 of the 752 sick foals showed an OD value of 0.3 or higher and were designated as 'suspected of R. equi infection'. Age distribution during the initial medical examination...
Redox regulation of large conductance Ca(2+)-activated K+ channels in smooth muscle cells. The effects of sulfhydryl reduction/oxidation on the gating of large-conductance, Ca(2+)-activated K+ (maxi-K) channels were examined in excised patches from tracheal myocytes. Channel activity was modified by sulfhydryl redox agents applied to the cytosolic surface, but not the extracellular surface, of membrane patches. Sulfhydryl reducing agents dithiothreitol, beta-mercaptoethanol, and GSH augmented, whereas sulfhydryl oxidizing agents diamide, thimerosal, and 2,2'-dithiodipyridine inhibited, channel activity in a concentration-dependent manner. Channel stimulation by reduction and inhibit...
Utilization of both phenotypic and molecular analyses to investigate an outbreak of multidrug-resistant Salmonella anatum in horses. Phenotypic and molecular techniques, including antimicrobial susceptibility testing, plasmid analysis, and pulsed-field gel electrophoresis (PFGE) were used to characterize 15 isolates of multidrug-resistant (MDR) Salmonella anatum cultured during a 16 mo period from horses and a veterinary clinic environment. The isolates were resistant to multiple antimicrobial agents and could be placed into 4 groups based on their antimicrobial resistance patterns. The isolates contained multiple plasmids ranging in size from 2 to > 100 kb that could be grouped into 3 different plasmid profile patterns;...
Comparative studies on mammalian Müller (retinal glial) cells. Müller cells from 22 mammalian species were subjected to morphological and electrophysiological studies. In the 'midperiphery' of retinae immunocytochemically labeled for vimentin, estimates of Müller cell densities per unit retinal surface area, and of neuron-to-(Müller) glia indices were performed. Müller cell densities were strikingly similar among the species studied (around 8000-11,000 mm-2) with the extremes of the horse ( or = 20,000 mm-2). By contrast, the number of neurons per Müller cell varied widely, being clustered at 6-8 (in retinae with many cones), at about 16, and at up t...
Diagnosis and management of a malignant granulosa cell tumor in a holstein nulligravida: a case study. Transrectal palpation of an 18-mo-old, anestrous Holstein nulligravida revealed an enlarged left ovary. Ultrasonographically, the mass resembled known equine granulosa cell tumor (GCT) images. Slowly, virilism developed. Fifty-five days after initial diagnosis, unilateral ovariectomy was performed The presurgical serum concentration of testosterone was equal to that of a mature bull. During the 3 mo period following surgery, estrus expression resumed, the serum testosterone concentration returned to baseline, and the heifer was diagnosed pregnant following AI. A low grade, malignant GCT was th...
Borna disease virus infection in animals and humans. The geographic distribution and host range of Borna disease (BD), a fatal neurologic disease of horses and sheep, are larger than previously thought. The etiologic agent, Borna disease virus (BDV), has been identified as an enveloped nonsegmented negative-strand RNA virus with unique properties of replication. Data indicate a high degree of genetic stability of BDV in its natural host, the horse. Studies in the Lewis rat have shown that BDV replication does not directly influence vital functions; rather, the disease is caused by a virus-induced T-cell mediated immune reaction. Because antibodi...
Localized sequence heterogeneity in the long terminal repeats of in vivo isolates of equine infectious anemia virus. The role of in vivo long terminal repeat (LTR) sequence variation of the lentivirus equine infectious anemia virus (EIAV) has not been explored. In this study, we investigated the heterogeneity found in the LTR sequences from seven EIAV-seropositive horses: three horses with clinical disease and four horses without any detectable signs of disease. LTR sequences were targeted in this study because the LTR U3 enhancer region of tissue culture-derived isolates has been identified as one of the few hypervariable regions of the EIAV genome. Furthermore, LTR variation may regulate EIAV expression in...
Transmission of some species of internal parasites in horses born in 1993, 1994, and 1995 on the same pasture on a farm in central Kentucky. Data are presented on the last 3 years of a 7-year study (1989-1995) on transmission of natural infections of internal parasites in horse foals (n = 27) born in 1993, 1994, and 1995 on the same pasture on a farm in central Kentucky. The foals were in a closed breeding herd of horses. Research on the first 4 years (1989-1992) of the study was published earlier (Lyons et al., 1991, 1994). Thirty-five species of endoparasites were identified, including 24 species of small strongyles. Monthly, seasonal, and host-age transmission patterns were elucidated for the parasites. Comparison of data betwee...
Cellular heterogeneity in cathepsin D distribution in equine articular cartilage. The distribution of cathepsin D in normal equine growth cartilage has been examined immunocytochemically using an antiserum raised against human cathepsin D. The cross-reactivity and specificity of the antiserum for equine cathepsin D was confirmed, and its lysosomal localisation was demonstrated in horse skin fibroblasts by confocal scanning microscopy. Cultured horse chondrocytes were heterogenous in their expression of cathepsin D. Heterogeneity of distribution of the enzyme was also seen in chondrocytes in cartilage from different anatomical sites. A high level of cathepsin D was observed ...
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