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Differential cell analysis and phenotypic subtyping of lymphocytes in bronchoalveolar lavage fluid from clinically normal dogs
1995
Vail, D.M. | Mahler, P.A. | Soergel, S.A.
In 33 healthy dogs, 66 bronchoalveolar lavage samples from the right and left caudal lung lobes were analyzed for volume of return, cellularity, differential cellularity, and immunophenotypic lymphocyte subpopulations. Lavage return was 64.8% (mean) following 3 sequential 25-ml lavages, for a total lavage volume of 75 ml. With this technique, 21.1 X 10(6) cells/sample (mean) were obtained. The cellular components of bronchoalveolar lavage samples, in decreasing order of frequency, were alveolar macrophages (79.4%), lymphocytes (13.5%), eosinophils (3.6%), mast cells (2.1%), epithelial cells (0.8%), and neutrophils (0.6%). Mean alveolar lymphocyte subpopulation frequencies, determined in 18 samples, for pan T, CD4, and CD8 cells were 52, 21.9, and 17.8%, respectively, with a CD4/CD8 ratio of 1.3. Variables analyzed did not vary between right and left caudal lung lobes, nor were they affected by body weight.
Показать больше [+] Меньше [-]Anatomic and radiographic appearance of a sesamoid bone in the tendon of origin of the supinator muscle of the cat
1995
Wood, A.K.W. | McCarthy, P.H. | Martin, I.C.A.
Results of radiologic and anatomic studies of each cubital articulation (elbow) of a group of 50 adult cat cadavers indicated that a sesamoid bone may be located in a constantly present sesamoid cartilage associated with the tendon of origin of the supinator muscle. Radiography revealed a sesamoid bone in 40 of the 100 tendons of origin of the supinator muscles dissected from the elbows. The sesamoid bone articulated with the craniolateral aspect of the head of the radius, and the larger sesamoid cartilage, which contained the bone, articulated with the head of the radius and the capitulum of the humerus. Of several possible functions of the sesamoid cartilage (bone), it was considered that protection of the craniolateral part of the humeroradial articulation and maintenance of the complex anatomic system during joint movement were important. In radiographic views of the elbows of lame cats, the sesamoid bone should not be mistaken for a chip fracture or an osteocartilaginous loose body.
Показать больше [+] Меньше [-]Detection of pseudorabies viral DNA in tonsillar epithelial cells of latently infected pigs
1995
Brown, T.T. Jr | Shin, K.O. | Fuller, F.J.
The Rice strain of pseudorabies virus (PRV) was intranasally instilled in pigs that were seronegative to PRV. Cells were scraped or brushed from tonsillar surfaces biweekly until pigs were euthanatized at either 10 or 16 weeks after infection. The DNA extracted from tonsillar cells or parenchyma were subjected to polymerase chain reaction analysis, using either a single set of oligonucleotide primers or nested primers from the PRV gII glycoprotein gene. Pigs became seropositive to PRV by 3 weeks after infection. The virus was isolated from the trigeminal ganglia and tonsils of pigs that were euthanatized or died 1 to 2 weeks after infection, but not from pigs that were euthanatized 10 or 16 weeks after infection. The PRV gene products were consistently detected in trigeminal ganglia and tonsils of all pigs at 1, 10, and 16 weeks after infection, and sporadically in the nasal mucosa, lymph nodes, and lungs of pigs that were euthanatized or died during the first 2 weeks after infection. Cells collected biweekly from tonsillar surfaces were mostly nucleated, squamous epithelial cells with fewer lymphocytes and neutrophils. Polymerase chain reaction analysis of DNA extracted from these cells revealed PRV DNA in a large proportion of the samples when sufficient cells were collected to provide 1 microgram of extracted DNA for use in the reaction mixtures. A second group of pigs had PRV strain 4892 intranasally instilled. The virus was isolated from tonsillar swab specimens until 3 weeks after infection. Tonsillar brushing specimens were collected biweekly until 14 weeks after infection. Some brushing specimens contained all nucleated, squamous epithelial cells, whereas other specimens contained a mixture of epithelial cells and up to 15% neutrophils, lymphocytes, and small mononuclear cells. Results of polymerase chain reaction analysis of DNA extracted from tonsillar cells collected 5, 11, and 14 weeks after infection were consistently positive for PRV gene products. Intact cells collected from tonsillar surfaces were placed in polymerase chain reaction mixtures with nested oligonucleotide primers from the PRV gII glycoprotein gene and were subjected to multiple amplification cycles. Afterward, the specificity of the amplified PRV gene products was determined by hybridization procedures, using a virus-specific oligonucleotide probe. Most nucleated, squamous epithelial cells stained positive for PRV DNA, suggesting that these cells were the primary source of PRV gene products in tonsillar brushing specimens.
Показать больше [+] Меньше [-]Effect of bethanechol, neostigmine, metoclopramide, and propranolol on myoelectric activity of the ileocecocolic area in cows
1995
Steiner, A. | Roussel, A.J. | Martig, J.
The effect of bethanechol, neostigmine, metoclopramide, and propranolol on myoelectric activity of the ileum, cecum, and proximal loop of the ascending colon was determined in 6 healthy Jersey cows implanted with 8 pairs of bipolar electrodes. Assigned at random, each cow received each of 5 treatments in 3-day intervals. The treatments included bethanechol (0.07 mg/kg of body weight, SC), neostigmine (0.02 mg/kg, SC), metoclopramide (0.15 mg/kg, IM), DL-propranolol (0.2 mg/kg, IM), and 0.9% sodium chloride (NaCl) solution (20 ml, SC). All drugs were administered during early phase I of the migrating myoelectric complex in the ileum. Myoelectric activity was recorded for 4 hours after treatment, and data were analyzed for each hour separately. Bethanechol and neostigmine significantly (P < 0.05) increased the number of cecocolic spikes per minute per electrode, duration of cecocolic spike activity (%), and number of cecocolic propagated spike sequences per 10 minutes, relative to NaCI, during 1 or more hours of the recording period. The effect of bethanechol was more pronounced on duration of spike activity and number of propagated spike sequences, whereas neostigmine mainly increased the number of (uncoordinated) spikes. Metoclopramide and propranolol had no significant effect on cecocolic myoelectric activity, relative to NaCl. It was concluded that bethanechol and, less likely, neostigmine at the dosage used in this study may be suitable for medical treatment of cecal dilatation in cattle in which hypomotility of the cecum and proximal loop of the ascending colon has to be reversed. The potential advantage of bethanechol vs neostigmine for medical treatment of cecal dilatation is worth further evaluation.
Показать больше [+] Меньше [-]Definition of chemiluminescence and superoxide production responses of bovine neutrophils to selected soluble and particulate stimulants, and comparisons with the responses to Pasteurella haemolytica
1995
Watson, G.L. | Slocombe, R.F. | Robinson, N.E. | Sleight, S.D.
We defined methods for use of luminol-dependent chemiluminescence (LDCL) and superoxide anion (O2-) production as parameters of the oxidative metabolism of neutrophils isolated from 1.5- to 5-week-old neonatal calves. We determined how variations in blood sample handling, agonist preparation, individual variability, and age of calves influenced the LDCL and O2- responses to certain agonists, and defined concentrations of soluble and particulate agonists that maximally stimulated the oxidative metabolism of bovine neutrophils. Oxidative responses, particularly LDCL, were characterized by marked dayto-day variability, differed greatly within and between calves, were partially age-dependent, and were partially dependent on the individual agonist. Superoxide anion production had substantially less variability. We compared the in vitro oxidative (LDCL and O2-) responses of neutrophils isolated from neonatal calves stimulated by defined concentrations of the agonists-latex, phorbol myristate acetate, calcium ionophore, and opsonized zymosan-with responses to formylated oligopeptides and zymosan-activated serum, and to live, dead, live opsonized, and dead opsonized Pasteurella haemolytica organisms. Opsonization of particulates, pathogenic or nonpathogenic, enhanced the LDCL and O2- responses of stimulated neutrophils although P haemolytica was a less potent stimulant of oxidative functions than were nonbiological agonists. We conclude that the generation of reactive oxygen species by bovine neutrophils in response to P haemolytica is highly dependent on the presence of opsonins and is greatly enhanced in live vs killed bacteria. Futhermore, the in vitro generation of reactive oxygen species, including O2- by stimulated neutrophils, may be of biologic importance if similar events occur in vivo, and could have a major role in the pathogenesis of the acute lung injury associated with pneumonic pasteurellosis.
Показать больше [+] Меньше [-]Development of monoclonal antibodies and capture immunoassays for feline immunodeficiency virus
1995
Dreitz, M.J. | Dow, S.W. | Fiscus, S.A. | Hoover, E.A.
We generated monoclonal antibodies (MAB) against feline immunodeficiency virus (FIV) and characterized these MAB by single competition enzyme immunoassays (EIA), immunoblot analysis, and radioimmunoprecipitation. Four MAB identified 3 distinct epitopes of the FIV p24/26 gag major core protein. One MAB recognized the p16/17 gag protein none recognized envelope proteins. We developed an FIV p26 antigen capture EIA that proved more sensitive (0.5 ng of p26/ml), less expensive, and less time-consuming than reverse transcriptase assay. The same MAB were used to develop an antibody EIA specific for FIV p26. The MAB and capture assays reported should prove useful in FIV diagnosis and research.
Показать больше [+] Меньше [-]Effects of alpha 2-adrenergic receptor agonist and antagonist drugs on cholinergic contraction in bovine tracheal smooth muscle in vitro
1995
Manning, M.M. | Broadstone, R.V.
Effects of alpha 2-adrenergic receptor stimulation on the cholinergic contractile response of bovine tracheal smooth muscle were studied. To determine the presence and function of alpha 2-adrenergic receptors on cholinergic nerves innervating bovine tracheal muscle, effects of 2 alpha 2-adrenoceptor agonists and an antagonist were determined. Muscular contractions were elicited by either electrical field stimulation (EFS) or exogenous acetylcholine (ACH). The contractile response to EFS and exogenous ACH was examined for each tissue. Electrical field stimulation of bovine tracheal smooth muscle caused contractions that were completely abolished by atropine, indicating the predominant excitatory innervation of bovine trachea is cholinergic. The alpha 2-adrenoceptor agonists clonidine and medetomidine (10(-6)M to 10(-4)M) concentration-dependently inhibited the contractile response to EFS but not the response to exogenous ACH. Contractions induced by EFS were significantly (P < 0.05) inhibited in clonidine (10(-4) M)-treated tissues at low frequencies (0.1 to 10 Hz), whereas medetomidine (10(-5)M, 10(-4)M) inhibited contractions at all frequencies (0.1 to 30 Hz). Inhibitory effects of the alpha 2-adrenoceptor agonists clonidine and medetomidine were attenuated by the alpha 2-adrenoceptor antagonist tolazoline. The alpha 2-agonists used in this study appear to cause prejunctional inhibition of cholinergic nerves, because the smooth muscle contractions elicited by EFS, but not exogenous ACH, were inhibited, compared with controls.
Показать больше [+] Меньше [-]Investigation of pseudorabies virus DNA and RNA in trigeminal ganglia and tonsil tissues of latently infected swine
1995
Cheung, A.K.
Pseudorabies virus (PRV) nucleic acids in the trigeminal ganglia and tonsils of swine latently infected with the virus were analyzed. By use of DNA-polymerase chain reaction (PCR), 14 of 14 trigeminal ganglia and 12 of 14 tonsils were positive for PRV genomes. By use of RNA-PCR, RNA containing the large latency transcript splice junction were detected in 4 of 4 trigeminal ganglia and 4 of 5 tonsils. In general, results of both PCR procedures indicated that the amounts of PRV DNA and RNA per microgram of cellular nucleic acids were higher in trigeminal ganglia than in tonsils. Identification of peripheral tissues that harbor latent PRV is an important asset for PRV research. The presence of large latency transcript in tonsil tissues, in the absence of virus replication, is a critical characteristic, which indicates that the tonsil is a site of PRV latency. For diagnostic purposes, animals need not be euthanatized to obtain their nervous tissue to determine latency; instead, tonsil biopsy specimens could be obtained from live animals for analysis. For pathogenesis studies, multiple specimens obtained sequentially from the same animal would be available for examination for the duration of the experiment.
Показать больше [+] Меньше [-]Comparison of argyrophilic nucleolar organizer regions by counting and image analysis in canine mammary tumors
1995
Destexhe, E. | Vanmanshoven, P. | Coignoul, F.
Two techniques for evaluating argyrophilic nucleolar organizer regions (AgNOR) were compared on 74 canine mammary tumors to discriminate between benign and malignant lesions. For each lesion, direct counting of AgNOR on at least 100 cell nuclei was compared with area, perimeter, and integrated optical density AgNOR dot values determined by image analysis. Significant differences between benign and malignant tumors were observed with both methods; however, lesions determined as aggressive or proliferative by histologic evaluation were only singled out by image analysis measurements. Image analysis, in our hands, was a reliable, precise, and convenient technique to characterize malignancy in canine mammary tumors.
Показать больше [+] Меньше [-]Efficacy of a variety of disinfectants against Actinobacillus pleuropneumoniae serotype 1
1995
Gutierrez, C.B. | Rodriguez Barbosa, J.I. | Suarez, J. | Gonzalez, O.R. | Tascon, R.I. | Rodriguez Ferri, E.F.
The efficacy of 23 disinfectants (including the most commonly used chemical groups) and 6 quaternary ammonium compound based commercial formulations against Actinobacillus pleuropneumoniae serotype 1 (ATCC 4074) was studied. The organisms were tested in suspension and carrier tests with serum as the organic matter. Chloramine-T, hydrogen peroxide, glutaraldehyde, and mercurochrome alone, and a quatemary ammonium compound formulation containing 10% benzalkonium chloride, 2.5% glutaraldehyde, 6.8% glyoxal, and 6% formaldehyde were effective in all tests, regardless of the presence or absence of organic load. All but 2 of the nonformulated disinfectants (sodium hypochlorite and an iodophor) caused at least a 3-log10 reduction in colony-forming units in the suspension test. However, most of the disinfectants were not as effective in the carrier test as in the suspension test; this difference ranged from a 1- to 5-log10 reduction in colony-forming units. In addition, the presence of serum considerably reduced the disinfectant capacities of most of the compounds tested, particularly in the carrier test. These results indicate the importance of selecting suitable disinfectants for routine use on surfaces contaminated with this organism, especially in the presence of organic matter. Chloramine-T and the aforementioned commercial formulation were also tested directly under field conditions in pig nurseries, confirming their high effectiveness.
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