Evaluation of L-selectin expression and assessment of protein tyrosine phosphorylation in bovine polymorphonuclear neutrophil leukocytes around parturition   总被引：3，自引：0，他引：3  

Monfardini E  Paape MJ  Wang Y  Capuco AV  Husheem M  Wood L  Burvenich C 《Veterinary research》2002,33(3):271-281

Impaired polymorphonuclear neutrophil leukocyte (PMN) function around parturition has been associated with increased clinical mastitis in dairy cows. Rolling and attachment of PMN to the endothelium is the first step in the recruitment process and is accomplished by interaction between L-selectin on PMN and its ligand on endothelial cells. Furthermore, tyrosine phosphorylation is involved in the initiation of many PMN functions. The objective of this work was to determine changes in expression of L-selectin and tyrosine phosphorylation in the perinatal period. Eight clinically healthy Holstein cows were used as PMN donors at d-21, -14, -7,0 (calving), +1, +2, +7, +14, +28. Evaluation of L-selectin expression was carried out on activated and resting PMN. Anti-bovine L-selectin monoclonal antibody (MAB) and flow cytometric analysis were used to measure the percentage of PMN fluorescing and receptor expression (log mean fluorescent channel, LMFC). Activated and resting PMN showed similar trends in % PMN fluorescence and LM FC. The percentage of PMN fluorescing tended to decrease at parturition, followed by a significant increase at d +14 and +28 (P < 0.02). For LMFC a decrease was observed on d +1 followed by an increase through d +28 (P < 0.01). Protein tyrosine phosphorylation of lysates prepared from PMN isolated throughout the study was detected by electrophoresis and western blotting using anti-phosphotyrosine MAB. Several protein bands were tyrosine phosphorylated. Two of these bands (42-44 kDa and 90 kDa) varied in intensity over time. The intensity of the 42-44 kDa band gradually increased from d -7, peaked at d +7 (P < 0.03), and steadily decreased to d +28 (P < 0.02). Antibody to activated mitogen protein kinase reacted with the 42-44 kDa band. Reduced PMN function during the periparturient period could be related to reduced L-selectin adhesion molecules on the cell surface, and to modulation in the phosphorylation of functionally important molecules.  相似文献   

Effect of intravenous infusion of recombinant ovine leptin on feed intake and serum concentrations of GH, LH, insulin, IGF-1, cortisol, and thyroxine in growing prepubertal ewe lambs   总被引：3，自引：0，他引：3  

Morrison CD  Wood R  McFadin EL  Whitley NC  Keisler DH 《Domestic animal endocrinology》2002,22(2):103-112

In sheep, serum concentrations of leptin change congruently with increases or decreases in nutritional status, while intracerebroventricular infusions of leptin dramatically suppress feed intake in well-fed lambs, and may also increase growth hormone (GH), and/or luteinizing hormone (LH) in undernourished lambs. The objective of the present study was to determine the effects of peripherally delivered ovine leptin, via intravenous infusions, on feed intake and serum concentrations of GH, LH, insulin, IGF-1, cortisol, and thyroxine. Twelve ewe lambs weighing 29.4 +/- 0.7 kg were infused intravenously with a linearly increasing dose of leptin or saline (n = 6 per group) for 10 days, reaching a maximum dose delivered of 0.5mg/h on day 10. Feed intake was assessed twice daily, and blood samples were collected every 10 min for 6 h on days 0, 2, 5, 8, and 10. Serum concentrations of leptin increased in leptin-treated lambs by day 2 (P = 0.05), and continued to increase to concentrations 9-fold greater than saline-infused lambs by day 10 (P < 0.001). Despite the substantial increase in serum leptin, feed intake did not differ between leptin and saline-infused lambs except on day 3.5 (P = 0.01). Furthermore, intravenous infusions of leptin did not significantly influence serum concentrations of insulin, cortisol, IGF-1, thyroxine, LH, or GH. Collectively, these observations contrast with the potent hypophagic effects of leptin when delivered intracerebroventricularly into well-fed lambs. The reasons for the disparate response of lambs treated intravenously with leptin, versus that reported for lambs treated intracerebroventricularly with leptin are not known, but may provide insight into the mechanism(s) of leptin resistance.  相似文献   

Flow cytometry: clinical applications in equine medicine     

Davis EG  Wilkerson MJ  Rush BR 《Journal of veterinary internal medicine / American College of Veterinary Internal Medicine》2002,16(4):404-410

The use of flow cytometry in veterinary diagnostics is becoming a valuable clinical tool with a broad range of applications. Physical characteristics of cells can be determined by the flow cytometer laser and electronics through the measurement of changes in light scatter properties. Other components and functions of cells can be defined through the application of fluorochrome dyes that have an affinity for cellular components. Traditionally, common clinical applications are immunophenotyping of cells of the hematopoietic system with fluorescent-labeled antibodies raised against specific cell surface proteins. Other approaches have been used to elucidate changes in cell function and DNA content. This review is intended to provide the reader with the fundamental uses of flow cytometry. Examples of clinical applications in equine patients include immune-mediated hemolytic anemia, immune-mediated thrombocytopenia (IMT), chronic inflammatory disease, and neoplasia.  相似文献   

Dermatophyte granulomas caused by Trichophyton mentagrophytes in a dog     

Bergman RL  Medleau L  Hnilica K  Howerth E 《Veterinary dermatology》2002,13(1):49-52

Multiple, dermal and subcutaneous nodules developed in a young female Manchester Terrier dog that had a chronic history of superficial dermatophytosis. Skin biopsy specimens of the nodules revealed granulomatous inflammation in the deep dermis and subcutis with branching fungal organisms. Cultures of multiple biopsy specimens from the nodules all yielded Trichophyton mentagrophytes. The lesions in this dog were similar to granulomatous dermatophytosis, a skin disease that has been reported in Persian cats and one Yorkshire Terrier dog.  相似文献   

Pheasant ataxia: a new condition in pheasant poults     

de B Welchman D  Ainsworth HL  Pennycott TW  MacKenzie G  Wood AM 《The Veterinary record》2000,147(4):93-97

Between 1995 and 1997 a neurological condition in pheasant poults from 24 sites in England and Scotland was investigated. Affected birds showed varying degrees of ataxia and incoordinated movements and, in severe cases, recumbency, but generally remained alert with their heads held upright. The condition characteristically affected poults from seven weeks of age and the incidence on any one site was low. No significant bacteria were isolated consistently from brain tissue. The condition was characterised histologically by a non-suppurative meningoencephalitis, in which lesions were found predominantly in the cerebellum in 61 of 81 samples examined (75.3 per cent). A non-suppurative myelitis was recorded in 16 of 20 spinal cords examined. No lesions were recorded in peripheral neural tissue and lesions were rare in other tissues. The condition appeared not to have been recorded previously in pheasants. A viral aetiology was suspected but Newcastle disease virus was not involved.  相似文献   

Characterisation of monoclonal antibodies to ovine interleukin-6 and the development of a sensitive capture ELISA     

McWaters P  Hurst L  Chaplin PJ  Collins RA  Wood PR  Scheerlinck JP 《Veterinary immunology and immunopathology》2000,73(2):155-165

A purified recombinant ovine (rOv) interleukin-6 (IL-6) was used to generate specific murine monoclonal antibodies (mAbs) and a polyclonal rabbit antisera to this cytokine. From the 31 initial hybridoma cell lines generated, three stable clones were established which secreted mAbs to rOvIL-6, as judged by a direct enzyme-linked immunosorbent assay (ELISA) and Western blotting. Their specificity was further confirmed by demonstrating that none of the mAbs recognised any of the six other irrelevant recombinant ovine cytokines tested by direct ELISA. All three mAbs displayed cross-reactivity with human and African green monkey IL-6 as demonstrated by direct ELISA and Western blotting. In contrast, the polyclonal antibodies only cross-reacted with bovine IL-6 and not with either of the human or monkey homologues. By combining a mAb with the polyclonal antisera a sensitive, IL-6-specific, capture ELISA was developed that had a sensitivity of 150 pg/ml. This detection system was unequivocally validated by demonstrating that native OvIL-6 could be detected in efferent lymph draining from a stimulated popliteal lymph node. In addition, one of the mAbs was shown to allow the detection of OvIL-6 by intracellular cytokine staining and flow cytometry.  相似文献   

Ultrasonography of the equine neonatal kidney     

Hoffmann KL  Wood AK  McCarthy PH 《Equine veterinary journal》2000,32(2):109-113

Ultrasonography is important in the clinical examination of the foal. The ultrasonographic appearance and size of the neonatal kidneys were defined and an imaging protocol established in 6 normal Thoroughbred foals (mean age +/- s.d. 5.0 +/- 3.2 days). Characteristically, in both the heart-shaped right kidney and bean-shaped left kidney, the renal cortex was more echogenic than the medulla. The terminal recesses, renal crest and pelvis were identified, as was the ureter, which contained anechoic urine in its lumen. The renal, interlobar and arcuate vessels were seen. For the right kidney, the ultrasonographic probe was placed at the 14-17th intercostal spaces and paralumbar fossa. For the left kidney, the probe was at the 16th or 17th intercostal spaces and paralumbar fossa. Perirenal structures, including the caudate lobe of the liver, the dorsal extremity of the spleen, the adrenals, the aorta and caudal vena cava were also identified. An understanding of the ultrasonographic appearance of the normal neonatal kidney, accompanied by a routine imaging protocol to ensure that all regions of each kidney are examined, permit a more informed interpretation of renal images in the first few days postpartum.  相似文献   

Experimental transmission of bovine anaplasmosis (caused by Anaplasma marginale) by means of Dermacentor variabilis and D. andersoni (Ixodidae) collected in western Canada          下载免费PDF全文

Murray W. Lankester  W. Brad Scandrett  Elizabeth J. Golsteyn-Thomas  Neil C. Chilton    Alvin A. Gajadhar 《Canadian journal of veterinary research》2007,71(4):271-277

Canadian cattle are free of bovine anaplasmosis, with the exception of 4 isolated incursions since 1968, which were eradicated. It is not known why the disease has not become established in regions of Canada adjacent to the United States where it is endemic. To assess the vector competence of wild-caught ticks in cattle-rearing regions, Dermacentor variabilis and D. andersoni were collected in western Canada and fed on calves experimentally infected with Anaplasma marginale (St. Maries strain). The 2 tick species were equally competent in transmitting A. marginale to splenectomized calves, all 15 tick-exposed calves becoming infected. The prepatent periods in 13 calves ranged from 18 to 26 d and did not vary in relation to the numbers of ticks fed or the duration of transmission feedings. The unusually long prepatent periods in 2 calves (45 and 55 d) were probably due to concomitant Eperythrozoon infection. This study clearly demonstrated that tick species present in western Canada are competent vectors of bovine anaplasmosis. Potential barriers, including climate, must be considered in developing strategies to prevent A. marginale from becoming established in anaplasmosis-free regions.  相似文献   

Graham Warner Ward     

E Tudor  A Wood 《Australian veterinary journal》2007,85(1-2):84-84

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Protocol: a highly sensitive RT-PCR method for detection and quantification of microRNAs   总被引：5，自引：0，他引：5  

Erika Varkonyi-Gasic  Rongmei Wu  Marion Wood  Eric F Walton  Roger P Hellens 《Plant methods》2007,3(1):12

MicroRNAs (miRNAs) are a class of small non-coding RNAs with a critical role in development and environmental responses. Efficient and reliable detection of miRNAs is an essential step towards understanding their roles in specific cells and tissues. However, gel-based assays currently used to detect miRNAs are very limited in terms of throughput, sensitivity and specificity. Here we provide protocols for detection and quantification of miRNAs by RT-PCR. We describe an end-point and real-time looped RT-PCR procedure and demonstrate detection of miRNAs from as little as 20 pg of plant tissue total RNA and from total RNA isolated from as little as 0.1 μl of phloem sap. In addition, we have developed an alternative real-time PCR assay that can further improve specificity when detecting low abundant miRNAs. Using this assay, we have demonstrated that miRNAs are differentially expressed in the phloem sap and the surrounding vascular tissue. This method enables fast, sensitive and specific miRNA expression profiling and is suitable for facilitation of high-throughput detection and quantification of miRNA expression.  相似文献