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Cytokine production during endotoxin-induced mastitis in lactating dairy cows.
1993
Shuster D.E. | Kehrli M.E. Jr. | Stevens M.G.
The role of interleukin-1 (IL-1), interleukin-6 (IL-6), and tumor necrosis factor a during endotoxin-induced mastitis in cows was characterized. Six cows had 10 microgram of Escherichia coli lipopolysaccharide infused into 1 mammary gland. Three other cows served as nontreated controls. Within 1.5 to 2.5 hours after infusion, endotoxin caused obvious edema of the mammary gland and increased serum albumin concentration in milk of infused glands 6 times. Milk somatic cell count began to increase 3 to 5 hours after infusion in all treated glands. At 7 hours after infusion, somatic cell counts were increased > 10 times, compared with counts in milk from control cows. Pyrexia of > 1 C developed in only 1 cow, but all treated cows had serum cortisol concentrations > 50 ng/ml in response to endotoxin treatment. High concentrations of IL-1 (10 to 600 U/ml) and IL-6 (2 to 22 U/ml) were detected in milk of infused glands beginning 2.5 to 4 hours after infusion. Endotoxin did not induce detectable amounts of tumor necrosis factor activity in milk or serum. Swelling and mammary gland permeability changes preceded any detectable increase in IL-1 and IL-6 activity, indicating that these clinical signs of inflammation were not mediated by these cytokines. Systemic responses and the leukocytic influx into endotoxin-infused glands developed after or concurrently with initial increases in IL-1 and IL-6 activities in milk. These results suggested that IL-1 and IL-6 may have a role in mammary gland defenses and in the pathophysiologic changes during endotoxin-induced mastitis.
显示更多 [+] 显示较少 [-]Immunohistochemical detection of porcine reproductive and respiratory syndrome virus using colloidal gold
1993
Magar, R. | Larochelle, R. | Robinson, Y. | Dubuc, C.
Two cytopathic agents were isolated on porcine alveolar macrophages following inoculation with homogenates of lung tissues from pigs showing respiratory problems. These isolates were identified as porcine reproductive and respiratory syndrome (PRRS) virus isolates by indirect immunofluorescence using a PRRS virus (PRRSV) specific monoclonal antibody (MAb) and were designated as LHVA-92-1 and LHVA-92-2. Immunogold electron microscopy using a porcine PRRS positive serum pool and protein A-gold resulted in an intense labelling of aggregates of viral particles. Dark specific cytoplasmic staining of porcine alveolar macrophages infected with both virus isolates could be observed by immunogold silver staining (IGSS) using the specific MAb. This method proved effective in detecting PRRSV antigens in several ethanol-fixed tissues of piglets intranasally inoculated with the supernatants of macrophages infected with each isolate. Immunogold silver staining was also successfully used for the detection of PRRSV antigens on sections of formalin-fixed paraffin-embedded lung tissues and on frozen sections of lungs. The present results indicate that colloidal gold may be useful for the identification and immunohistochemical detection of PRRSV in tissues.
显示更多 [+] 显示较少 [-]Inhibition of myelopoiesis by serum from dogs exposed to estrogen
1993
The mechanism of estrogen-induced myelotoxicosis is unknown, although evidence indicates that estrogen does not directly damage the bone marrow granulocyte-macrophage progenitor cells and that the thymus is a probable mediator of the bone marrow suppression. Estrogen-induced production of a myelopoiesis-inhibitory factor by canine thymic stromal cells in vitro has been observed. Then, presence of a myelopoiesis-inhibitory factor in canine serum was investigated immediately after estrogen administration in vivo. Maximal reduction in colony-forming units-granulocyte/macrophage growth by sera from individual dogs varied. Individual dog sensitivity to estrogen-induced myelotoxicosis is seen clinically, and the cause is unknown. This serum factor could have a role in the eventual bone marrow hypoplasia seen in estrogen-treated dogs and is possibly the same factor produced by cultured thymic stromal cells exposed to estrogen.
显示更多 [+] 显示较少 [-]Recognition of triiodothyronine-containing epitopes in canine thyroglobulin by circulating thyroglobulin autoantibodies
1993
Gaschen, F. | Thompson, J. | Beale, K. | Keisling, K.
Serum triiodothyronine autoantibody (T3 AA), triiodothyronine (T3), and thyroxine (T4) concentrations were determined in 45 canine sera containing substantial amounts of thyroglobulin autoantibodies (Tg AA); sera also were assayed to investigate the ability of free T3 to inhibit Tg AA binding to canine Tg. Serum T3 AA concentrations defined 2 groups of sera; 28 sera had low T3 AA concentration (less than or equal to 20 ng/ml) and 17 sera had high T3 AA concentration (greater than or equal to 250 ng/ml). Direct linear correlation between T3 AA concentration and apparent serum T3 concentration was observed (r = 0.75). Serum with low T3 AA concentration had apparent T3 concentration that was significantly (P < 0.01) lower than that in serum with high T3 AA concentration. Mean serum T4 concentration was not significantly different between serum with low or high T3 AA concentration. Mean Tg AA activity was significantly (P < 0.05) lower in serum with low T3 AA concentration than in serum with high T3 AA concentration. Addition of free T3 to serum significantly (P < 0.05) decreased detectable activity of Tg AA in both groups of sera. However, significant difference in magnitude of the reduction was not observed between sera with low or high T3 AA concentration. Results indicate that a fraction of Tg AA recognizes T3-containing epitopes in Tg. Increased prevalence of T3 AA for serum with high Tg AA activity indicates that T3 AA may be another valid indicator of lymphocytic thyroiditis. These antibodies may be generated against the hormonogenic epitopes of Tg.
显示更多 [+] 显示较少 [-]Cytokine production during endotoxin-induced mastitis in lactating dairy cows
1993
Shuster, D.E. | Kehrli, M.E. Jr | Stevens, M.G.
The role of interleukin-1 (IL-1), interleukin-6 (IL-6), and tumor necrosis factor a during endotoxin-induced mastitis in cows was characterized. Six cows had 10 microgram of Escherichia coli lipopolysaccharide infused into 1 mammary gland. Three other cows served as nontreated controls. Within 1.5 to 2.5 hours after infusion, endotoxin caused obvious edema of the mammary gland and increased serum albumin concentration in milk of infused glands 6 times. Milk somatic cell count began to increase 3 to 5 hours after infusion in all treated glands. At 7 hours after infusion, somatic cell counts were increased > 10 times, compared with counts in milk from control cows. Pyrexia of > 1 C developed in only 1 cow, but all treated cows had serum cortisol concentrations > 50 ng/ml in response to endotoxin treatment. High concentrations of IL-1 (10 to 600 U/ml) and IL-6 (2 to 22 U/ml) were detected in milk of infused glands beginning 2.5 to 4 hours after infusion. Endotoxin did not induce detectable amounts of tumor necrosis factor activity in milk or serum. Swelling and mammary gland permeability changes preceded any detectable increase in IL-1 and IL-6 activity, indicating that these clinical signs of inflammation were not mediated by these cytokines. Systemic responses and the leukocytic influx into endotoxin-infused glands developed after or concurrently with initial increases in IL-1 and IL-6 activities in milk. These results suggested that IL-1 and IL-6 may have a role in mammary gland defenses and in the pathophysiologic changes during endotoxin-induced mastitis.
显示更多 [+] 显示较少 [-]Pharmacokinetics and effects of repeated administration of phenylbutazone in neonatal calves
1993
Semrad, S.D. | McClure, J.T. | Sams, R.A. | Kaminski, L.M.
Age, species, and disease state may substantially alter the disposition and clearance of pharmacologic agents. This is particularly important when drugs with low therapeutic index are used in ill neonates. Pharmacokinetic variables for phenylbutazone were determined in 24- to 32-hour-old healthy and endotoxemic calves after IV administration of a single dose (5 mg/kg of body weight, IV). Elimination halflife was 207 and 168 hours, and clearance was 0.708 and 0.828 ml/kg/h in healthy and endotoxemic calves, respectively. Intravenous infusion of endotoxin at the dose (2 micrograms/kg over 4 hours) given did not significantly alter any of the calculated pharmacokinetic variables. Serum thromboxane B2 concentration was significantly (P = 0.05) suppressed for 3 hours after phenylbutazone administration in healthy calves and for 4 hours in endotoxin-challenged calves. Daily administration of phenylbutazone (10 mg/kg loading, then 5 mg/kg for 9 days) to healthy and endotoxemic calves failed to induce any lesions consistent with nonsteroidal anti-inflammatory drug toxicosis.
显示更多 [+] 显示较少 [-]3 alpha-hydroxylated bile acid profiles in clinically normal cats, cats with severe hepatic lipidosis, and cats with complete extrahepatic bile duct occlusion
1993
Center, S.A. | Thompson, M. | Guida, L.
Concentrations of 3 alpha-hydroxylated bile acids were measured in serum and urine of clinically normal (healthy) cats (n = 6), cats with severe hepatic lipidosis (n = 9), and cats with complete bile duct occlusion (n = 4). Bile acid concentrations were measured by use of a gradient flow high-performance liquid chromatography procedure with an acetonitrile and ammonium phosphate mobile phase and an in-line postanalytic column containing 3 alpha-hydroxysteroid dehydrogenase and a fluorescence detector. Specific identification of all bile acid peaks was not completed; unidentified moieties were represented in terms of their elution time (in minutes). Significant differences in serum and urine bile acid concentrations, quantitative and proportional, were determined among groups of cats. Cats with hepatic lipidosis and bile duct occlusion had significantly (P greater than or equal to 0.05) greater total serum and urine bile acids concentrations than did healthy cats. The proportion of hydrophobic bile acids in serum, those eluting at greater than or equal to 400 minutes, was 1.9% for healthy cats, 3.3% for cats with lipidosis, and 5.4% for bile duct-obstructed cats. Both groups of ill cats had a broader spectrum of unidentified late-eluting serum bile acids than did healthy cats; the largest spectrum developed in bile duct-occluded cats. The trihydroxy-to-dihydroxy serum bile acids ratio was 8.8:1 for healthy cats; 24.1:1 for cats with lipidosis: and 20:1 for cats with bile duct obstruction. There was a paucity of glycine-conjugated bile acids in all cats and small quantities of secondary bile acids in ill cats. A significantly (P < 0.05) smaller proportion of unconjugated primary bile acids was detected in sera from ill cats. Serum taurolithocholic acid was detected only in small quantities in cats of each group. There was significantly increased quantity, but lower proportion, of trihydroxy-cholestanoic acid in serum from ill cats, compared with healthy cats. A significantly (P < 0.05) greater proportional amount of unidentified moieties eluting at 130 and 277 minutes was detected in urine of cats with hepatic lipidosis; we believe that the unidentified moiety eluting at 277 minutes is tau- tauroallocholic acid. Large proportional amounts of taurocholic and cholic acids were detected in urine of all cats, but ill cats had significantly (P < 0.05) greater quantities (quantitatively and proportional). Ill cats had significantly (P < 0.05) more taurocholic than cholic acid in urine. Because taurine is an essential amino acid for cats and is a necessary daily dietary constituent, large urinary losses of taurine in conjugated bile acids may further compromise the health of anorectic cats with severe hepatic lipidosis.
显示更多 [+] 显示较少 [-]Effects of sample collection and handling on concentration of osteocalcin in equine serum
1993
Hope, E. | Johnston, S.D. | Hegstad, R.L. | Geor, R.J. | Murphy, M.J.
A commercially available radioimmunoassay kit for measurement of human osteocalcin was validated for use in horses. For accurate measurement of equine serum osteocalcin, blood samples may be collected at a temperature between 20 and 25 C, then centrifuged within 90 minutes; serum may be stored at - 20 C in plastic tubes for up to 26 weeks. Serum may be thawed and refrozen up to 5 times without significant change in measured equine serum osteocalcin concentration. Assay sensitivity was 0.16 ng/ ml. Recovery of bovine osteocalcin standard added to equine serum was linear. Intra-assay coefficient of variation (x 100) for 2 equine serum pools was 6.9 (mean +/- SD, 13.9 +/- 1.0 ng/ml) and 7.5 (10.6 +/- 0.8 ng/ml) %. Interassay coefficient of variation for 3 equine serum pools measured in 12 assays was 12.5 (16.1 +/- 2.0 ng/ml), 12.7 (11.5 +/- 1.5 ng/ml), and 24.6 (3.0 +/- 0.7 ng/ml) %. Dilutional parallelism was documented by assaying pooled equine serum at 4 dilutions and correcting the mean result for dilution. Significant change was not observed in equine serum osteocalcin concentration for various time-of-day blood sample collections in horses housed under continuous lighting.
显示更多 [+] 显示较少 [-]The use of intradermal carrageenan in calves to estimate the dose of oxindanac, a nonsteroidal anti-inflammatory drug
1993
King, J. N.
A simple and humane model of inflammation, induced by the intradermal injection of 0.3 mL of sterile 2% carrageenan, was characterized in calves by measuring the volume of skin swelling plus histological analysis of skin biopsies. Carrageenan produced a biphasic increase in skin swelling, with an early edematous response followed by a more chronic cellular infiltrate. The swelling and sensitivity to pressure observed in the early response were suitable for testing the antiedematous and analgesic activity of a new nonsteroidal anti-inflammatory drug (NSAID), oxindanac. Pretreatment with intravenous oxindanac at doses from 0.5 to 8.0 mg/kg reduced the volume of swelling and this reached statistical significance (p < 0.05) at 2 mg/kg. The ED50 and ED90 values for inhibition of the peak swelling volume (4 h) were estimated to be 1 mg/kg and 2 mg/kg, respectively. These compare with an ED90 of 2.0 mg/kg for inhibition of serum TxB2 production, an index of platelet cyclo-oxygenase activity. The dose of oxindanac required for antiedematous activity correlated, therefore, with maximal inhibition of serum TxB2. The analgesic activity of oxindanac reached no clear maximum response, but statistically significant difference (p < 0.05) from placebo was reached with doses of 2 mg/kg and above. It is concluded that intradermal carrageenan produced a simple, humane and useful model for dose estimation of a new NSAID in calves.
显示更多 [+] 显示较少 [-]Selective parathyroidectomy of the dog
1993
Finco, D. R. | Brown, S. A. | Ferguson, D. C. | Crowell, W. A.
Selective parathyroidectomy (PTX) is preferred to thyroparathyroidectomy (TPTX) when specific effects of parathyroid hormone depletion are being studied. However, because of the anatomic proximity of thyroid and parathyroid glands, TPTX often is performed, leaving animals depleted of thyroxine (T4) and calcitonin as well as parathyroid hormone (PTH). In the present study, six normal dogs had parathyroid tissue and about seven-eighths of thyroid tissue removed. This quantity of thyroid tissue was inadequate to maintain normal serum T4 concentrations, despite allowance of 168 days for thyroid recovery. Five of six dogs with reduced renal mass had successful selective PTX and normal serum T4 concentrations at 28 days, when one-half or more of thyroid tissue was spared. We conclude that with attention to the surgical technique, selective PTX can be achieved in a high percentage of dogs and sufficient thyroid tissue spared to maintain euthyroidism.
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