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Cytologic evaluation of bronchoalveolar lavage fluid obtained from Standardbred racehorses with inflammatory airway disease.
1995
Moore B.R. | Krakowka S. | Robertson J.T. | Cummins J.M.
Cytologic examination of bronchoalveolar lavage fluid (BALF), including phenotypic analysis of lymphocytes, was performed on 32 Standardbreds with poor race performance and endoscopic examination findings characteristic of inflammatory airway disease (IAD). Nucleated cell counts in BALF from IAD-affected horses were higher than those in control horses; the cytologic profile of BALF in affected horses included mixed inflammation, characterized by mild neutrophilia, lymphocytosis, and monocytosis. Eosinophil and mast cell counts were not higher in the IAD-affected group, compared with those in the control group; however, 4 IAD-affected horses had marked eosinophilia (24.7 +/- 4.8% SEM) in BALF. Phenotypic analysis of lymphocytes in BALF obtained from IAD-affected horses revealed a low proportion of CD4-positive cells and B cells, compared with those in the control group; these findings may have been representative of a greater proportion of non-B, non-T cells (null cells) in horses with IAD. The cytologic profile of BALF obtained from horses with IAD differed from that in horses affected with chronic obstructive pulmonary disease, suggesting that the pathogenesis of inflammation in horses with IAD may differ from that of chronic obstructive pulmonary disease.
Mostrar más [+] Menos [-]Cytologic and bacteriologic evaluation of tracheobronchial aspirates from clinically normal foals.
1989
Crane S.A. | Ziemer E.L. | Sweeney C.R.
Thirty-eight tracheobronchial aspirates (TBA) were collected from twenty 1 to 6-month-old foals, which were free of clinical signs of respiratory tract or other infectious disease. We collected TBA from 9 of the foals 3 times when they were approximately 8, 16, and 24 weeks old. Aspirates were examined cytologically after staining with modified Wright-Giemsa, Gram, toluidine blue, and prussian blue stains. Aerobic bacterial culturing was performed on all aspirates. Of the 20 initial TBA, 4 (20%) were normal cytologically on the basis of previously defined criteria for TBA from clinically normal horses, 6 (30%) had a high percentage of eosinophils (> 5%), 8 (40%) were classified as indicative of subacute inflammation, and 2 (10%) were classified as indicative of acute inflammation. Nine (45%) were positive for mast cells and none were positive for hemosiderin-laden macrophages (hemosiderophages). Of the 9 foals from which samples were collected at 16 and 24 weeks of age, results were similar, except for an increase in the number of TBA classified as indicative of chronic inflammation (33% and 22% respectively) and the number positive for hemosiderophages (33% and 88%, respectively). One TBA was considered nondiagnostic because of pharyngeal contamination. Culturing of 12 of the 37 aspirates (32%) yielded a potential microbial pathogen. Only 2 were positive cultures from the same foal. The following organisms were isolated: beta-hemolytic Streptococci spp (4), Actinobacillus/Pasteurella spp (4), Rhodococcus equi (2), unidentified nonenteric Gram-negative rod (1), and Escherichia coli (1). Thirty-four of the 37 aspirates (92%) yielded light growth of various organisms considered to be nonpathogenic and normal inhabitants of the upper respiratory tract. It was concluded that the presence of inflammatory cells, eosinophils, and mast cells in the tracheobronchial aspirates from clinically normal foals is a common finding. These cytologic findings were consistent in the samples collected from foals at 8, 16, and 24 weeks of age. It was also concluded that bacteria with recognized pathogenicity can be isolated from TBA from clinically normal foals and were most frequently isolated from 1- to 2-month-old foals or those with cytologic evidence of inflammation, even in the absence of clinical signs of respiratory tract disease.
Mostrar más [+] Menos [-]Activity of feline interferon-omega after ocular or oral administration in cats as indicated by Mx protein expression in conjunctival and white blood cells
2006
Bracklein, T. | Theise, S. | Metzler, A. | Spiess, B.M. | Richter, M.
Objective-To assess the biological response to recombinant feline interferon-omega (rFeIFN-omega) following ocular or oral administration in cats via estimation of Mx protein expression in conjunctival cells (CCs) and WBCs. Animals-10 specific pathogen-free cats. Procedures-In multiple single-dose drug experiments, each cat received various concentrations of rFeIFN-omega administered topically into both eyes (50 to 10,000 U/eye) and orally (200 to 20,000 units). The same cats received saline (0.9% NaCl) solution topically and orally as control treatments. The CCs and WBCs were collected prior to treatment (day 0), on day 1, and every third or seventh day thereafter until samples yielded negative results for Mx protein. Samples were examined for Mx protein expression via immunohistochemistry and immunoblotting procedures involving murine anti-Mx protein monoclonal antibody M143. Results-After topical application of 10,000 U of rFeIFN-omega/eye, CCs stained for Mx protein for a minimum of 7 days, whereas WBCs were positive for Mx protein for a minimum of 31 days. After topical application of lower concentrations, CCs did not express Mx protein, in contrast to WBCs, which stained for Mx protein at 1,000 units for at least 1 day. Following oral administration, Mx protein was expressed in WBCs at rFeIFN-omega concentrations as low as 200 units, whereas CCs did not stain for Mx protein at any concentration. Conclusions and Clinical Relevance-Results indicate that Mx protein expression (a marker of the biological response to rFeIFN-omega) in CCs and WBCs of rFeIFN-omega-treated cats depends on the dose of rFeIFN-omega, site of administration, and cell type.
Mostrar más [+] Menos [-]Effect of exercise on activation of the p38 mitogen-activated protein kinase pathway, c-Jun NH2 terminal kinase, and heat shock protein 27 in equine skeletal muscle
2006
Ginneken, M.M.E van | Graaf-Roelfsema, E de | Keizer, H.A. | Dam, K.G van | Wijnberg, I.D. | Kolk, J.H van der | Breda, E van
Objective-To investigate the effects of exercise on activation of mitogen-activated protein kinase (MAPK) signaling proteins in horses. Animals-6 young trained Standardbred geldings. Procedure-Horses performed a 20-minute bout of exercise on a treadmill at 80% of maximal heart rate. Muscle biopsy specimens were obtained from the vastus lateralis and pectoralis descendens muscles before and after exercise. Amount of expression and intracellular location of phosphospecific MAPK pathway intermediates were determined by use of western blotting and immunofluorescence staining. Results-Exercise resulted in a significant increase in phosphorylation of p38 pathway intermediates, c-Jun NH2 terminal kinase (JNK), and heat shock protein 27 (HSP27) in the vastus lateralis muscle, whereas no significant changes were found in phosphorylation of extracellular regulated kinase. In the pectoralis descendens muscle, phosphorylation of p38 and HSP27 was significantly increased after exercise. Immunohistochemical analysis revealed fiber-type-specific locations of phosphorylated JNK in type 2a/b intermediate and 2b fibers and phosphorylated p38 in type 1 fibers. Phosphorylated HSP27 was strongly increased after exercise in type 1 and 2a fibers. Conclusions and Clinical Relevance-The p38 pathway and JNK are activated in the vastus lateralis muscle after a single 20-minute bout of submaximal exercise in trained horses. Phosphorylation of HSP27 as detected in the study reported here is most likely induced through the p38 signaling pathway.
Mostrar más [+] Menos [-]Evaluation of porcine ileum models of enterocyte infection by Lawsonia intracellularis
2006
McOrist, S. | Gebhart, C.J. | Bosworth, B.T.
The early interaction of Lawsonia intracellularis with host cells was examined with the use of porcine ileum models. Two conventional swine were anesthetized, and ligated ileum loops were prepared during abdominal surgery. The loops were inoculated with 10⁸ L. intracellularis or saline. After 60 min, samples of each loop were processed for routine histologic and electron microscopic study. Histologic and ultrathin sections of all the loops appeared normal, with no apposition of bacteria and host cells or bacterial entry events in any loop. Portions of ileum from a single gnotobiotic piglet were introduced as xenografts into the subcutis of each flank of 5 weaned mice with severe combined immunodeficiency disease. After 4 wk, 10⁸ L. intracellularis were inoculated into each of 4 viable xenografts with a sterile needle; the other 3 viable xenografts received saline. Histologic and ultrathin sections of all the xenografts 3 wk after inoculation showed relatively normal porcine intestinal architecture, with normal crypts, crypt cell differentiation, and low villous structures; the xenografts treated with the bacteria also showed intracytoplasmic L. intracellularis within crypt and villous epithelial cells. Thus, entry of L. intracellularis into target epithelial cells and multiplication may not be sufficient alone to directly cause cell proliferation. A proliferative response may require active division of crypt cells and differentiation in conjunction with L. intracellularis growth.
Mostrar más [+] Menos [-]New approach to percutaneous muscle biopsy in dogs
1995
Reynolds, A.J. | Fuhrer, L. | Valentine, B.A. | Kallfelz, F.A.
The size and quality of muscle specimens obtained by use of a percutaneous biopsy technique were studied. All biopsies were performed under local anesthesia, using an 11-gauge biopsy needle. The mean +/- SEM size of specimens obtained from 128 biopsies of the semitendinosus muscles of 16 Alaskan Huskies was 23.8 +/- 4.4 mg. All biopsy specimens were of sufficient quality to permit histochemical differentiation of the fiber types by use of myosin ATPase staining. An additional 8 biopsy specimens were obtained from 1 dog and analyzed for muscle glycogen content. These specimens contained 50.6 +/- 7.2 mmol of glucose/kg of muscle wet weight. This modified biopsy procedure was free of notable complications, and repeatable use produced specimens of adequate size and quality for histologic and biochemical analysis. It is concluded that this procedure is a safe and reliable alternative to open biopsy for diagnosis and management of neuromuscular, metabolic, and nutritional myopathies.
Mostrar más [+] Menos [-]Comparative analyses of peritoneal fluid from calves and adult cattle
1995
Anderson, D.E. | Cornwell, D. | Anderson, L.S. | St-Jean, G. | Desrochers, A.
Reference values for hematologic variables change with increasing age in cattle. Therefore, the purpose of the study reported here was to describe the peritoneal fluid constitutents of clinically normal young calves, and to compare cellular concentration and distribution in blood and peritoneal fluid of young calves with those of adult cattle. Eight healthy 8-week-old male Holstein calves and 8 healthy 3- to 8-year-old Holstein cows were studied. Peritoneal fluid was collected from calves along the ventral midline, 4-cm cranial to the umbilicus. Abdominocentesis was performed in the region of the lower right flank in adult cattle. Correlation analysis, using the Pearson's correlation coefficient, and regression analysis were performed for blood and peritoneal fluid data from calves. Data from calves were compared with those of cows, using Wilcoxon's rank sum test. A P value < 0.05 was considered significant for all tests. Calves had significantly lower blood eosinophil count (P < 0.003) and plasma protein concentration (P < 0.001) than did cows. Calves had significantly higher peritoneal fluid nucleated cell (P < 0.05) and mononuclear cell (P < 0.05) counts, but lower peritoneal fluid eosinophil cell count (P < 0.003) than did cows. For calves, nulceated cell and lyhocyte cell counts in the blood had a high, positive correlation with those of peritoneal fluid. However, the prediction equation for nucleated cell count accounted for a modest proportion of variability. A prediction equation for peritoneal fluid lymphocyte cell count was established. On the basis of results of this study, reference ranges established for peritoneal fluid constituents of clinically normal adult cattle may not be appropriate for interpretation of peritoneal fluid analysis of calves.
Mostrar más [+] Menos [-]Effect of coculture with stallion spermatozoa on de novo protein synthesis and secretion by equine oviduct epithelial cells
1995
Thomas, P.G.A. | Ignotz, G.G. | Ball, B.A. | Brinsko, S.P. | Currie, W.B.
Adhesion of equine spermatozoa to homologous oviduct epithelial cells (OEC) in vitro results in specific changes in spermatozoa and OEC function. To test the hypothesis that adhesion of spermatozoa affects protein synthesis and secretion by OEC, the following treatment groups were established in culture: OEC with culture medium only; control spermatozoa in culture medium only; OEC in coculture with spermatozoa; and OEC and spermatozoa in coculture, but physically separated by a microporous membrane. The experiment was replicated within each of 4 ejaculates from 3 stallions. De novo protein secretion by OEC was measured and compared by incorporation of [35S]methionine, and evaluated, using two-dimensional polyacrylamide gel electrophoresis and fluorography. Monolayers of OEC secreted a large number of proteins of molecular mass ranging from 14 to 205 kd. Adhesion of spermatozoa consistently caused reduced synthesis of 2 OEC secretory proteins and new or increased synthesis of 6 proteins. When spermatozoa and OEC were separated by a microporous membrane, some but not all of these changes were duplicated. Synthesis of 3 OEC secretory proteins, unaffected by binding of spermatozoa, was reduced when spermatozoa were prevented from contact with OEC by a microporous membrane. Adhesion of equine spermatozoa to homologous OEC monolayers and presence of equine spermatozoa resulted in qualitative and quantitative changes in synthesis and secretion of proteins by OEC. These changes have implications for storage, longevity, and maturation of spermatozoa.
Mostrar más [+] Menos [-]Detection of bovine immunodeficiency virus in blood and milk-derived leukocytes by use of polymerase chain reaction
1995
Nash, J.W. | Hanson, L.A. | St Cyr Coats, K.
Bovine immunodeficiency virus (BIV) is prevalent in beef and dairy cattle, yet the mode(s) of BIV transmission are undefined. Using polymerase chain reaction, which specifically targeted a 235-bp, highly conserved region of the BIV pol gene, BIV-infected leukocytes were detected in the blood and milk of BIV-seropositive cows. These data confirm the presence of BIV in milk and identify the potential for lactogenic transmission of this virus.
Mostrar más [+] Menos [-]Cytologic characterization of bronchoalveolar lavage fluid collected through an endotracheal tube in cats
1994
Hawkins, E.C. | Kennedy-Stoskopf, S. | Levy, J. | Meuten, D.J. | Cullins, L. | DeNicola, D. | Tompkins, W.A.F. | Tompkins, M.B.
Bronchoalveolar lavage was performed through an endotracheal tube in 34 specific-pathogen-free cats to determine expected values for bronchoalveolar lavage fluid cytologic analysis, using this method of collection. Saline solution for lavage was instilled in 3 separate aliquots at a volume of 5 ml/kg of body weight each. Analysis of sequential aliquots was performed to investigate the differences in cell counts among the 3 fractions. The effect of combining aliquots, including or omitting the first fraction, was evaluated to determine whether all aliquots could be combined for analysis without substantially affecting results. The total number of nucleated cells retrieved from each cat ranged from 0.9 to 31.1 X 10(6). Most of these cells were macrophages (78 +/- 15%, mean +/- SD) and eosinophils (16 +/- 14%). The first aliquot had the greatest number of epithelial cells, and the lowest total nucleated cell count and relative and absolute eosinophil counts. Differences among aliquots also were identified for relative and absolute macrophage counts, relative and absolute neutrophil counts, and absolute lymphocyte count. Statistically significant differences were found for many of the cell counts when values from the combination of the second and third aliquots were compared with values from the combination of all 3 aliquots. Magnitude of the differences was small, and these differences were not believed to be of practical consequence.
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