Refinar búsqueda
Resultados 1-10 de 12
Complete primary sequence of equine cartilage link protein deduced from complementary DNA
1995
Dudhia, J. | Platt, D.
Investigation of the structure of equine articular cartilage link protein (LP) from individuals ranging in age from 1 to 15 years identified 3 distinct isoforms having molecular weights of 46,000, 43,000, and 41,000. The relative amounts of each of the 3 isoforms altered with age. The largest form did not change with age; however, amounts of the Mr 43,000 and 41,000 forms increased with increasing age. The results suggested that an accumulation, in the extracellular matrix of cartilage, of these 2 smaller products may have arisen from proteolytic cleavage. The complete amino acid sequence of the protein core was determined from complementary DNA products prepared by polymerase chain reaction amplification of cartilage LP mRNA. The sequence had 96% similarity with human LP and with that of other species for which the primary structure has been determined. This high degree of sequence conservation and the isoform data indicate that extracellular processing of LP occurs by similar mechanisms in various species. At the transcription level, equine chondrocytes were found to express LP as 2 abundant mRNA of 5.0 and 3.0 kb, and a smaller mRNA of 1.5 kb. Processing of the LP mRNA in horses, thus, appears to be similar to that found in other species investigated, and although multiple transcripts are present, the coding region remains unaltered and only 1 protein product is made.
Mostrar más [+] Menos [-]Evidence for endothelium-derived relaxing factor/nitric oxide in equine digital arteries
1995
Cogswell, A.M. | Johnson, P.J. | Adams, H.R.
Objective--To test the hypothesis that endothelium-derived nitric oxide modulates vasomotor reactivity in equine digital arteries. Design--Digital arteries were isolated from adult horses, and their vasodilator properties were examined in an in vitro controlled environment. Animals--Five adult horses (1 gelding, 4 mares) without evidence of hoof or vascular disease were studied. Procedure--Arterial rings with or without endothelium were exposed to endothelium-dependent vasodilator drugs in the presence or absence of a pharmacologic inhibitor of the enzyme nitric oxide synthase. Results--Vasodilator effects of 3 endothelium-dependent vasorelaxant agents were significantly greater in endothelium-intact vessels than in endothelium-denuded vessels. Moreover a nitric oxide synthase inhibitor reduced vasodilator responses to endothelium-dependent vasodilators in endothelium-intact arteries, but had no discernable effects in endothelium-denuded arteries. Conclusions--These findings indicate the presence of endothelium-derived relaxing factor/nitric oxide in blood vessels of horses, and identify vascular endothelium as an endogenous modulator of vasomotor tone in the digital arteries of this species.
Mostrar más [+] Menos [-]Potential for oxytetracycline administration by three routes to cause milk residues in lactating cows, as detected by radioimmunoassay (Charm II) and high performance liquid chromatography test methods
1995
Anderson, K.L. | Moats, W.A. | Rushing, J.E. | Wesen, D.P. | Papich, M.G.
Milk antimicrobial residues are a serious concern for the dairy industry. Residues of the tetracycline family of antimicrobials have been reported in market milk by investigators, using radioimmunoassay and microbial receptor technology (hereafter referred to as the Charm II test). In response to these reports, an investigation was conducted to determine the potential of 3 extra-label routes of oxytetracycline (OTC) administration to cause milk residues above the Food and Drug Administration safe value of 30 parts per billion (ppb). Lactating Holstein cows were administered OTC once by use of 1 of 3 routes: IV at 16.5 mg/kg of body weight (n = 6); IM at 11 mg/kg (n = 6); and intrauterine (IU) at 2 g in 500 ml of saline solution/cow (n = 6). Duplicate milk samples were collected at the milking prior to drug administration and for the next 13 milkings at 12-hour intervals. Concentrations of OTC in milk samples were analyzed by use of the Charm II test for tetracyclines (limit of OTC detection, approx 5 ppb) and were compared with concentrations determined by use of a high-performance liquid chromatography (HPLC) method (lower limit of OTC quantitation, approx 2 ppb). The potential for milk OTC residues above the Food and Drug Administration safe value of 30 ppb after treatment was considerably greater for the IV and IM routes, compared with the IU route. Mean peak OTC concentrations in milk at the first milking after treatment for the HPLC and Charm II tests were approximately 3,700 to 4,200 ppb for the IV route, 2,200 to 2,600 ppb for the IM route, and 186 to 192 ppb for the IU route, respectively. Pharmacokinetic analysis, based on milk OTC concentrations, indicated that the area under the curve (AUC) and milk maximal concentration (Cmax) differed significantly (P < 0.001) among routes of administration. The AUC was similar for IV and IM administrations; values for both were greater than the AUC for IU administration. The Cmax was greatest for IV, intermediate for IM, and least for IU administration. There were significant (P less than or equal to 0.01) differences in AUC between assay methods (Charm II vs HPLC) for the IV route. Concentrations of OTC in milk determined by the Charm II test were often greater than those determined by HPLC. Administration of OTC to lactating cows via these routes is extra-label drug use. Failure to withhold the product from early milkings of cows administered OTC by the IV or IM route should be considered a potential cause of OTC residues in market milk. Milk from nearly all cows contained OTC (< 30 ppb), the Food and Drug Administration safe level, by 120 hours after OTC administration. Use of appropriate withholding times and antibiotic residue testing is indicated to avoid OTC residues.
Mostrar más [+] Menos [-]Comparison of a radioimmunoassay (Charm II) test with high-performance liquid chromatography for detection of oxytetracycline residues in milk samples from lactating cattle
1995
Moats, W.A. | Anderson, K.L. | Rushing, J.E. | Wesen, D.P.
A radioimmunoassay test for tetracyclines (Charm II) was compared with high-pressure liquid chromatography (HPLC) for detection of oxytetracycline (OTC) residues in milk samples from individual lactating cows. Oxytetracycline was administered by 1 of 3 routes (IV, IM, or intrauterine) to 21 lactating dairy cows. A total of 292 duplicate milk samples were collected from milkings before and through 156 hours after OTC administration. Concentration of OTC in these samples was determined by use of the Charm II test and an HPLC method with a lower limit of quantitation, approximately 2 ng of OTC/ml. Samples were also classified with respect to presence of OTC residues relative to the FDA safe concentration (less than or equal to 30 ng/ml), using the Charm II (by control point determination) and HPLC methods. There was a significant (P less than or equal to 0.05) difference between test methods in classification of milk samples with respect to presence or absence of OTC at the FDA safe concentration. A total of 48 of the 292 test results (16.4%) did not agree. Using the HPLC test results as the standard with which Charm II test results were compared, 47 false presumptive-violative test results and 1 false presumptive-nonviolative Charm II test result (a sample containing 31 ng of OTC/ml, as evaluated by HPLC) were obtained. The samples with false presumptive-violative Charm II results contained (less than or equal to 30 ng of OTC/ml, as evaluated by HPLC. In some respects, the Charm II test performed appropriately as a screening test to detect OTC residues in milk samples from individual cows. However, the tendency for the test to yield presumptive-violative test results at OTC concentrations lower than the FDA safe concentration (as evaluated by HPLC), suggests that caution should be exercised in using the test as the sole basis on which a decision is made to reject milk. As indicated by the manufacturer, presumptive-violative Charm II test results should be confirmed by additional testing Although not specifically evaluated, the tendency for misclassification of milk samples as presumptive-violative by the Charm II test may or may not occur in commingled milk, compared with milk samples from individual cows.
Mostrar más [+] Menos [-]Subcellular biochemical changes during the development of the small intestine of pony foals
1995
Batt, R.M. | Burgess, D. | Edwards, G.B. | Waal, S. van de | Sorensen, S.H. | Hart, C.A.
To examine the postnatal development of equine small intestine, biopsy specimens of jejunal mucosa from 8 ponies, between 6 and 28 weeks old, were subjected to analytical subcellular fractionation and assay of organelle marker enzymes. Fractionation revealed a reduction in the particulate brush border component of beta-galactosidase (lactase) activity between 6 and 28 weeks, and a corresponding increase in soluble activity, although the reduction in mean specific activity was not significant. There also was a decrease in the proportion of brush border to soluble aminopeptidase N activity, a relative loss of brush border gamma-glutamyltransferase activity, and a considerable decrease in the specific activity of alkaline phosphatase throughout the gradient fractions. In contrast, there were marked increases in activities of (alpha-glucosidase (maltase) and sucrase in the older ponies, accompanied by considerable changes in the intracellular distribution of particulate alpha-glucosidase activity, which was predominantly associated with endoplasmic reticulum at 6 weeks, whereas the large increase in activity observed by 28 weeks was clearly associated with the brush border. The modal density of brush borders also increased with age, suggestive of an increase in the glycoprotein-to-lipid ratio of the microvillar membrane. In contrast to these brush border changes, there was relatively little alteration in the activities or density distributions of marker enzymes for endoplasmic reticulum, basolateral membranes, mitochondria, or lysosomes. These findings indicate that maturation of equine intestinal epithelium during the first few months of life results in major changes in the properties and enzyme composition of enterocyte brush borders.
Mostrar más [+] Menos [-]Administration of recombinant human interleukin 1 receptor antagonist during endotoxin-induced mastitis in cows
1995
Shuster, D.E. | Kehrli, M.E. Jr
The role of interleukin 1 (IL-1) as an inflammatory mediator during mastitis and the therapeutic effect of recombinant human IL-1 receptor antagonist (IL-1ra) for bovine mastitis was studied. Cows were intramammarily infused with lipopolysaccharide (25 g) in 1 mammary gland. Half the cows also received infusions of 5 mg of IL-1ra into the same mammary gland just prior to endotoxin infusion and 4, 8, and 12 hours later. After endotoxin infusion, tumor necrosis factor and high IL-1 bioactivity were detected in whey from infused glands. Vascular permeability changes and neutrophil accumulation in milk paralleled the appearance of cytokines. A systemic reaction, characterized by pyrexia and an increase in blood cortisol concentration, also were observed. Milk yield was inhibited and milk composition was altered in infused and noninfused glands. The increase in IL-1 bioactivity in milk after endotoxin infusion was almost completely prevented in glands receiving IL-1ra. However, IL-1ra had no effect on local inflammation, systemic reaction, or impairment in productive performance. These results indicate that IL-1 does not mediate its effect within the milk compartment, and suggest either that IL-1 is not critical to the mastitic response or that intramammary infusion of IL-1ra does not place the antagonist where IL-1 interacts with its receptor.
Mostrar más [+] Menos [-]Effect of psyllium on plasma concentration of glucose, breath hydrogen concentration, and fecal composition in calves with diarrhea treated orally with electrolyte solutions
1995
Naylor, J.M. | Liebel, T.
We investigated the effect of adding psyllium to a standard electrolyte solution in 10 calves with diarrhea. The calves were tested with the standard solution on one day and standard solution plus psyllium on the alternate day. The order of treatments was randomized. Psyllium converted the solution into mucilage, but did not affect fecal consistency. Mean +/- SEM area under the glucose absorption curve was lower for mucilaginous than for nonmucilaginous solutions, 2.1 +/- 0.62 vs 3.75 +/- 1.18 mmol.h, respectively, but the difference was not significant. The area under the breath hydrogen curve was marginally lower for mucilaginous than nonmucilaginous solutions, 102 +/- 20 and 209 +/- 60 ppm.h, respectively. The usefulness of such decreased bacterial fermentation is doubtful.
Mostrar más [+] Menos [-]Morphologic and morphometric characterization of lung collagen content in clinically normal adult Thoroughbreds in race training
1995
Lakritz, J. | Wisner, E.R. | Finucane, T. | O'Brien, T.R. | Tyler, W.S. | Pascoe, J.R. | Plopper, C.G.
Interstitial and bronchointerstitial pulmonary patterns are commonly observed in thoracic radiographs of Thoroughbreds. Prominent interstitial and bronchointerstitial pulmonary patterns are observed in clinically normal horses, and in horses with respiratory tract disease. Until recently, the relevance of these pulmonary patterns was not known. Previous studies indicated that bronchiolitis, bronchiolar epithelial hyperplasia, epithelial metaplasia, and bronchial arteriolar recruitment correlated strongly with the prominence of the interstitial and bronchointerstitial pulmonary patterns observed radiographically. We examined the content and distribution of collagen in the lungs of 7 clinically normal Thoroughbreds in race training. After standardized fixation, lung tissue was treated with a compound that selectively stains collagen. Standard morphometric techniques were used to determine the volume density of parenchymal tissue and parenchymal airspace, mean linear intercept (estimate of alveolar size), alveolar surface area-to-volume ratio, percentage of parenchyma composed of collagen, percentage of airway wall composed of collagen, and airway wall thickness. These values were compared with radiographic and histopathologic scores obtained from the same horses. The volume density of parenchymal tissue and small airway wall thickness correlated strongly with the prominence of the bronchial and bronchointerstitial pulmonary patterns observed radiographically. Small airway thickness was also highly correlated with the perceived prominence of the interstitial pulmonary patterns observed radiographically, and morphometric estimates of parenchymal tissue and parenchymal collagen. There were also strong correlations between the volume density of parenchymal tissue, the percentage of parenchymal collagen, peribronchiolar mononuclear cell infiltrates, and bronchiolar mucosal plication estimates. In horses with more prominent bronchiolar mucosal plication, there was a strong direct relation to the observed prominence of peribronchiolar and submucosal blood vessels, and the bronchial and bronchointerstitial patterns observed radiographically. Horses with prominent peribronchiolar mononuclear cell infiltrates also had more obvious interstitial and bronchointerstitial pulmonary patterns observed radiographically. There also was a direct correlation between the percentage of parenchymal collagen and the observed prominence of peribronchiolar and submucosal blood vessels in these horses. In all horses, there was a strong negative correlation between the estimated average alveolar size and the observed severity of the vascular and bronchial patterns observed radiographically. Four horses with the greatest estimated airway wall and interalveolar collagen had more prominent interstitial and bronchointerstitial densities and histopathologic evidence of bronchiolitis. These horses had evidence of epithelial basement membrane disruption, with disorganized collagen fibers running between the adventitial layer and the epithelial basement membrane. Amounts of collagen were greater in the adventitia and interalveolar septa, with the fibers appearing larger and more coarse and disorganized. In horses with the greatest percentage of interalveolar septal collagen, accumulations of collagen were larger in the interalveolar septal tips. These findings suggest that horses with prominent interstitial and bronchointerstitial pulmonary patterns radiographically have undergone previous episodes of pulmonary injury, which has resulted in deposition of increased amounts of collagen in interalveolar septa and airway walls.
Mostrar más [+] Menos [-]Histochemical and morphometric study of fiber types in ten skeletal muscles of healthy young adult cats
1995
Braund, K.G. | Amling, K.A. | Mehta, J.R. | Steiss, J.E. | Scholz, C.
A histochemical and morphometric study of fiber types in a variety of skeletal muscles of healthy young adult cats was undertaken to provide normative data not available previously. Using a standardized system of nomenclature, fiber types 1, 2A, 2B, and 2C were identified in most cat muscles on the basis of myosin ATPase staining at pH 4.45. Type-2M fibers were present in temporalis (TEM) and masseter (MAS) muscles. Type-1 fibers predominated in medial head of triceps (MHT) and soleus muscles. Type-2B fibers were dominant in biceps femoris, lateral head of gastrocnemius, cranial tibial, long head of triceps, and superficial digital flexor muscles; type-2A fibers were dominant in buccinator muscle samples; and type-2M fibers were dominant in TEM and MAS muscles. Numbers of type-2C fibers did not exceed 2 to 3% of the myofiber population in any muscle. In CT and LHT muscles, a gradient of fiber type distribution was observed, with significant (P < 0.05) increase in numbers of type-1 and type-2A fibers in deeper regions of the muscles. The distribution of fiber types was compartmentalized in MHT and MAS specimens. Diameter of type-2B fibers was significantly (P < 0.05) larger than that of type-1 and type-2A fibers in biceps femoris, lateral head of gastrocnemius, cranial tibial, long head of triceps, and superficial MHT muscles. Diameter of type-2M fibers was significantly (P < 0.05) larger than that of type-1 fibers in TEM and MAS muscles. The soleus type-1 muscle fibers were the largest fibers encountered in any muscle. In MHT muscle, fiber diameter of type-1 and type-2B fibers varied significantly (P < 0.05) in oxidative and glycolytic compartments. Variability coefficients were less than 200 in all muscles. In every muscle specimen, the number of fibers with internal nuclei was less than or equal to 2%.
Mostrar más [+] Menos [-]Bovine immunodeficiency virus in stud bull semen
1995
Bovine immunodeficiency virus (BIV), a lentivirus, is prevalent in dairy and beef cattle in southeastern United States and may be associated with a lymphoproliferative disease. The mode(s) of BIV transmission are undefined. Because artificial insemination is a common practice in dairy production, contaminated stud semen could serve as an important source of infection if the virus is harbored in seminal fluids. To evaluate this possibility, we procured 11 cryopreserved semen specimens from a stud semen repository. Leukocytes were purified from the specimens, and the leukocyte DNA was used as template in a polymerase chain reaction procedure that targeted a 235-base pair, highly conserved domain of the BIV pol gene. The target sequence was amplified from the seminal leukocyte DNA of 9 of the specimens (82%), and nucleotide sequencing confirmed the BIV specificity of the fragment. This finding provides evidence that stud bull semen may serve as an important reservoir of BIV, suggesting the possibility that artificial insemination of dairy cows may have a major role in transmission and wide-spread dissemination of this bovine lentivirus.
Mostrar más [+] Menos [-]