Objective—To demonstrate efficacy of flow cytometric evaluation of expression and activity of P-glycoprotein (P-gp) and multidrug resistance–associated protein (MRP) efflux pumps and characterize and correlate their expression and activity in grossly normal canine nodal lymphocytes.
Sample Population—Nodal lymphocytes from 21 clinically normal dogs.
Procedures—Pump expression was assessed by use of fluorescent-labeled mouse antihuman P-gp (C494) and MRP1 (MRPm6) antibodies and expressed as median values (antibody value divided by isotype control value). The P-gp and MRP activities were assessed by measuring cellular retention of rhodamine 123 and 5(6)-carboxyfluorescein diacetate in the absence and presence of inhibitors (verapamil and PSC833 for P-gp, probenecid and MK-571 for MRP). Protein activity was expressed as median fluorescence of cells with inhibitors divided by that without inhibitors.
Results—Expression of P-gp was (mean ± SEM) 50.62 ± 13.39 (n = 21) and that of MRP was 2.16 ± 0.25 (13). Functional activity was 1.27 ± 0.06 (n = 21) for P-gp and both inhibitors and 21.85 ± 4.09 (21) for MRP and both inhibitors. Function and expression were not correlated.
Conclusions and Clinical Relevance—Use of flow cytometry effectively assessed P-gp and MRP expression and activity in canine lymphocytes. Optimization of the flow cytometric assay was determined for evaluating activity and expression of these pumps in canine lymphoid cells. Evaluation of expression or activity may offer more meaning when correlated with clinical outcome of dogs with lymphoproliferative diseases. Cell overexpression of P-gp and MRP can convey drug resistance.
Objective—To compare plasma and synovial fluid
endothelin-1 (ET-1) and nitric oxide (NO) concentrations
in clinically normal horses and horses with joint
Animals—36 horses with joint disease, and 15 horses
without joint disease.
Procedure—Horses with joint disease were assigned
to 1 of the 3 groups (ie, synovitis, degenerative joint
disease [DJD], or joint sepsis groups) on the basis of
findings on clinical and radiographic examination and
synovial fluid analysis. Endothelin-1 and NO concentrations
were measured in plasma from blood samples,
collected from the jugular vein and ipsilateral
cephalic or saphenous vein of the limb with an affected
or unaffected joint, as well as in synovial fluid samples
obtained via arthrocentesis from the involved
Results—Plasma ET-1 concentrations between
affected and unaffected groups were not significantly
different. Median concentration and concentration
range of ET-1 in synovial fluid obtained from the joint
sepsis group (35.830 pg/mL, 7.926 to 86.614 pg/mL;
n = 7) were significantly greater than values from the
synovitis (17.531 pg/mL, 0.01 to 46.908 pg/mL; 18),
DJD (22.858 pg/mL, 0.01 to 49.990 pg/mL; 10), and
unaffected (10.547 pg/mL, 0.01 to 35.927 pg/mL; 10)
groups. Plasma and synovial fluid NO concentrations
between affected and unaffected groups were not
Conclusions and Clinical Relevance—Endothelin-1
is locally synthesized in the joints of horses with various
types of joint disease. Synovial fluid concentrations
of ET-1 varied among horses with joint disease,
with concentrations significantly higher in the synovial
fluid of horses with joint sepsis. These results
indicate that ET-1 may play a role in the pathophysiologic
mechanism of joint disease in horses.
(Am J Vet Res 2002;63:1648–1654)
Objective—To evaluate the effect of diets enriched with eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) on in vivo production of interleukin (IL)-1, IL-6, tumor necrosis factor (TNF)-α, prostaglandin E2 (PGE2), and platelet-activating factor (PAF) in dogs.
Animals—15 young healthy dogs.
Procedures—Dogs were randomly allocated to receive an isocaloric ration supplemented with sunflower oil (n = 5), fish oil (5), or fish oil plus vitamin E (5) for 12 weeks. At week 12, in vivo production of inflammatory mediators was evaluated in serum at multiple time points for 6 hours following stimulation with IV administration of lipopolysaccharide (LPS).
Results—Serum activity or concentration (area under the curve) of IL-1, IL-6, and PGE2 significantly increased after LPS injection in all groups but to a lesser extent in dogs receiving the fish oil diet, compared with results for dogs receiving the sunflower oil diet. Serum activity of TNF-α and PAF concentration also increased significantly after LPS injection in all groups but did not differ significantly among groups.
Conclusions and Clinical Relevance—A fish oil–enriched diet consisting of 1.75 g of EPA/kg of diet and 2.2 g of DHA/kg of diet (dry-matter basis) with an n-6:n-3 fatty acid ratio of 3.4:1 was associated with significant reductions in serum PGE2 concentrations and IL-1 and IL-6 activities. Results supported the use of EPA- and DHA-enriched diets as part of antiinflammatory treatments for dogs with chronic inflammatory diseases. Additional studies in affected dogs are warranted to further evaluate beneficial anti-inflammatory effects of EPA- and DHA-enriched diets.
Objective—To quantify changes in endothelium-derived factors and relate those changes to various aspects of digital hemodynamics during the prodromal stages of carbohydrate overload (CHO)-induced laminitis in horses.
Animals—20 adult horses without abnormalities of the digit.
Procedures—Digital and jugular venous blood samples were collected at 1-hour intervals (for assessment of endothelin-1 [ET-1] immunoreactivity and measurement of glucose, insulin, and nitric oxide [NO] concentrations) or 4-hour intervals (CBC and platelet-neutrophil aggregate assessment) for 8 hours or 16 hours after induction of CHO-associated laminitis in horses treated with an ET-1 antagonist. Effects of treatment, collection site, and time and the random effects of horse on each variable were analyzed by use of a repeated-measures model. Where treatment and collection site had no significant effect, data were combined.
Results—Compared with baseline values, CHO resulted in changes in several variables, including a significant increase from baseline in digital blood ET-like immunoreactivity at 11 hours; digital blood ET-like immunoreactivity was significantly greater than that in jugular venous blood at 8, 9, 11, and 12 hours. Digital and jugular venous blood concentrations of glucose increased from baseline significantly at 3, 4, and 5 hours; insulin concentration increased significantly at 5 hours; and the number of platelet-neutrophil aggregates increased significantly at 12 hours.
Conclusions and Clinical Relevance—In horses, concurrent increases in venous blood ET-1 immunoreactivity, insulin and glucose concentrations, and platelet-neutrophil aggregates support a role of endothelial dysfunction in the pathogenesis of CHO-induced laminitis.
Case Description—An 8-year-old domestic shorthair cat was evaluated because of signs of depression, circling, and visual deficits.
Clinical Findings—The cat had no cutaneous lesions, and results of an ophthalmologic examination and thoracic radiography were within reference limits. Computed tomography of the brain revealed a mass lesion involving the right parietal, temporal, and occipital lobes; the mass was in broad-based contact with the skull and smoothly marginated and had strong homogenous enhancement after contrast agent administration. During craniectomy, samples of the mass were collected for cytologic and histopathologic evaluations and microbial culture. A diagnosis of Blastomyces dermatitidis—associated meningoencephalitis with secondary pyogranulomatous inflammation was made.
Treatment and Outcome—Amphotericin B (0.25 mg/kg [0.11 mg/lb], IV) was administered on alternate days (cumulative dose, 1.75 mg/kg [0.8 mg/lb]). To minimize the risk of nephrotoxicosis, assessments of serum biochemical variables (urea nitrogen and creatinine concentrations) and urinalyses were performed at intervals. The third dose of amphotericin B was postponed 48 hours because the cat became azotemic. The cat subsequently received fluconazole (10 mg/kg [4.5 mg/lb], PO, q 12 h) for 5.5 months. Six months after discontinuation of that treatment, the cat appeared healthy and had no signs of relapse.
Clinical Relevance—Brain infection with B dermatitidis is typically associated with widespread disseminated disease. The cat of this report had no evidence of systemic disease. Blastomycosis of the CNS should be considered as a differential diagnosis for brain lesions in cats from areas in which B dermatitidis is endemic.
To evaluate survival times for dogs with previously untreated, peripheral nodal, intermediate- or large-cell lymphoma treated with prednisone alone.
109 client-owned dogs recruited from 15 institutions in the United States.
Dogs were treated with prednisone at a dosage of 40 mg/m2, PO, once daily for 7 days and at a dosage of 20 mg/m2, PO, once daily thereafter. Quality of life (QOL) was assessed by owners with a visual analog scale when treatment was started (day 0), 1 and 2 weeks after treatment was started, and every 4 weeks thereafter. The primary outcome of interest was survival time as determined by the Kaplan-Meier method. Factors potentially associated with survival time were examined.
Median overall survival time was 50 days (95% CI, 41 to 59 days). Factors associated with survival time included substage (a vs b) and immunopheno-type (B cell vs T cell). Owner-assigned QOL scores on days 0 and 14 were significantly positively correlated with survival time. When QOL score was dichotomized, dogs with day 0 or day 14 QOL scores ≥ 50 had significantly longer survival times, compared with dogs with day 0 or day 14 QOL scores < 50. No variables were predictive of long-term (> 120 days) survival.
CONCLUSIONS AND CLINICAL RELEVANCE
Results suggested that survival times were short for dogs with previously untreated, peripheral nodal, intermediate- or large-cell lymphoma treated with prednisone alone. Owner-perceived QOL and clinician-assigned sub-stage were both associated with survival time. Findings provide potentially important information for clinicians to discuss with owners of dogs with lymphoma at the time treatment decisions are made. (J Am Vet Med Assoc 2021;259:62–71)