Cytochemical staining characteristics of lymph nodes from normal and lymphoma-affected dogs

Frozen sections and imprint smears were used to evaluate the presence and pattern of cytochemical staining reactions in the B- and T-cell regions of lymph nodes from normal dogs and dogs with lymphoma. Staining procedures evaluated included peroxidase (PER), Sudan black B (SBB), naphthol AS-D chloroacetate esterase (CAE), alpha-naphthyl butyrate esterase (NBE), acid phosphatase (ACP), and leukocyte alkaline phosphatase (LAP). In normal lymph nodes, macrophages and some lymphocytes within the interfollicular (T-cell) region and medulla stained positive with ACP and NBE. Smaller numbers of macrophages also occurred sporadically within the germinal follicles. Cells positive for PER, SBB, and CAE were scattered infrequently throughout all regions of the normal lymph node, consistent with granulocytes and mast cells. The LAP stained cells were predominantly and prominently located within the mantle zone of secondary follicles and to a much lesser extent within the germinal centers, compatible with B-cell lymphocytes derived from follicular center cells. Of the 12 dogs with lymphoma, 7 cases (4 immunoblastic, 2 large noncleaved, 1 small noncleaved) stained diffusely positive with LAP, 4 cases (all lymphoblastic) had numerous focally positive lymphocytes using ACP and NBE, and 1 case (immunoblastic) did not stain positive with any of the cytochemical reactions. Cytochemical staining of canine lymph nodes with NBE, ACP, and LAP proved useful in distinguishing between B- or T-cell regions and detecting different cell types of canine lymphoma.     

Different biological behaviour of Waldenström macroglobulinemia in two dogs

Waldenström Macroglobulinemia is a low‐grade immunosecretory disorder associated with lymphoid tumours, which is rarely reported in veterinary medicine. In this study, we describe two clinical cases of this rare syndrome in dogs, each characterized by a different onset and clinical course. In one case, a hyperacute onset and aggressive behaviour of the neoplasm was observed. Absolute serum viscosity (SV) was retrospectively evaluated in order to explain clinical findings. Rotational viscosimetry showed good precision in measuring SV. Both dogs had SV values higher than a control groups of healthy dogs although only one subject developed hyperviscosity symptoms and complications. At high paraprotein concentrations, a slight reduction of the M‐component was associated with a marked decrease in SV. Thus, this work suggests that SV assessment is a relevant tool for managing monoclonal gammopathies, whose usefulness should be further confirmed in larger cohorts of dogs.     

Expression of P-glycoprotein and breast cancer resistance protein in three cases of canine lymphoma showing drug resistance

In canine lymphoma, drug resistance is the major factor hindering treatment. In this study, we performed immunohistochemical examination of P-glycoprotein (P-gp) and breast cancer resistance protein (BCRP), which are considered as transporters related to multidrug resistance in three recurrent canine lymphomas. All cases were negative for both transporters before anticancer drug administration, but became positive after this administration. The expression was confirmed in capillary endothelial cells, such as in brain capillaries acting as the blood-brain barrier (BBB). It is suggested that both transporters expressed on capillary endothelial cells in lymphoma tissue may inhibit the spread of anticancer drugs into tumor tissues from blood, the same as the BBB. Therefore, capillary endothelial cells could act as a blood-tumor barrier, which might be involved in drug resistance in canine lymphoma.     

Fractionated oral dosing and its effect on cyclophosphamide pharmacokinetics in dogs with high-grade multicentric lymphoma

Cyclophosphamide (CP) is an alkylating agent commonly included in multi-drug treatment protocols for canine cancer. As a prodrug, CP requires hepatic metabolism for activation to the intermediate compound 4-hydroxycyclophosphamide (4-OHCP) which then spontaneously forms alkylating phosphoramide mustard. CP is frequently administered in a fractionated manner, with the total dose given over multiple days. CP is reported to cause auto-induction of metabolism in humans, with faster CP clearance and relatively increased 4-OHCP formation following fractionated versus bolus dosing, however canine pharmacokinetic studies of CP dose fractionation are lacking. The study objective was to evaluate the pharmacokinetics of fractionated oral CP dosing at a dose of 200–250 mg/m² over 3 to 4 days in a prospectively identified population of cancer-bearing dogs. Plasma concentrations of CP and 4-OHCP were measured by ultra-high performance liquid chromatography tandem-mass spectrometry in eight dogs following the first and last doses to assess for auto-induction of CP metabolism. No significant difference in the rate of CP elimination between first and last doses were detected (0.73 ± 0.46 vs. 1.22 ± 0.5 h⁻¹; p = .125). Additionally, no significant difference in dose-normalized 4-OHCP exposure was identified between first and last doses (5.9 ± 2.1 vs. 7.9 ± 6.4 h × ng/ml; p = .936). These results suggest that fractionated dosing may not increase exposure to the active metabolite of CP in dogs as it does in humans. As such, standard bolus dosing and fractionated dosing may be equivalent in terms of bio-activation of CP in dogs administered a dose of 200–250 mg/m².