Dog erythrocyte antigens 1.1, 1.2, 3, 4, 7, and Dal blood typing and cross‐matching by gel column technique

Background: Testing for canine blood types other than dog erythrocyte antigen 1.1 (DEA 1.1) is controversial and complicated by reagent availability and methodology. Objectives: The objectives of this study were to use available gel column technology to develop an extended blood‐typing method using polyclonal reagents for DEA 1.1, 1.2, 3, 4, 7, and Dal and to assess the use of gel columns for cross‐matching. Methods: Dogs (43–75) were typed for DEA 1.1, 1.2, 3, 4, 7, and Dal. Methods included tube agglutination (Tube) using polyclonal reagents, a commercially available DEA 1.1 gel column test kit (Standard‐Gel) using monoclonal reagent, and multiple gel columns (Extended‐Gel) using polyclonal reagents. Blood from 10 recipient and 15 donor dogs was typed as described above and cross‐matched using the gel column technique. Results: Of 43 dogs typed for DEA 1.1, 23, 25, and 20 dogs were positive using Standard‐Gel, Extended‐Gel, and Tube, respectively. Typing for DEA 1.2 was not achievable with Extended‐Gel. For 75 dogs typed for DEA 3, 4, and 7, concordance of Extended‐Gel with Tube was 94.7%, 100%, and 84%, respectively. Dal, determined only by Extended‐Gel, was positive for all dogs. Post‐transfusion major cross‐matches were incompatible in 10 of 14 pairings, but none were associated with demonstrable blood type incompatibilities. Conclusions: Gel column methodology can be adapted for use with polyclonal reagents for detecting DEA 1.1, 3, 4, 7, and Dal. Agglutination reactions are similar between Extended‐Gel and Tube, but are more easily interpreted with Extended‐Gel. When using gel columns for cross‐matching, incompatible blood cross‐matches can be detected following sensitization by transfusion, although in this study incompatibilities associated with any tested DEA or Dal antigens were not found.     

Intra- and interlaboratory variability of allergen-specific IgE levels in atopic dogs in three different laboratories using the Fc-ɛ receptor testing

The IgE specific Fc-? receptor test is widely used for serological evaluation of allergic canine patients. Aim of the study was to evaluate intra- and interlaboratory variability in three independent laboratories. Duplicate serum aliquots from 15 atopic dogs were submitted simultaneously to 3 laboratories (LA, LB, LC), which subsequently performed the test in a blinded fashion. LA and LC analysed sera for 35 allergens and expressed the results as optical density units (OD), while LB analysed 15 allergens and provided reaction grades (RG). Results were compared with three factorial ANOVA. Intralaboratory variability was evaluated by calculating the dispersion factor for data of LA and LC and standard deviation (SD) for LB. Correlation coefficients (r) were used to evaluate interlaboratory variability. Depending on the allergens, intralaboratory dispersion factor for LA and LC varied from 1.15 to 6.63 and 1.19 to 6.17, respectively. In LB the SD varied between 0 and 0.6 RG. Regarding interlaboratory variability, correlation of OD values between LA and LC was excellent (r > 0.8, p < 0.001) for 13 of 35 allergens. There was a significant difference in OD results for 9 allergens. The results were significantly correlated (r > 0.8, p < 0.001) for all except 3 allergens (Dermatophagoides pteronyssinus, mixed grasses, nettle). As far as negative/positive results were concerned, intralaboratory differences were 3.14% and interlaboratory differences were 4.76%.     

Retrospective evaluation of episodic collapse in the horse in a referred population: 25 cases (1995-2009)

Background: Episodic collapse in horses has equine welfare and human safety implications. There are, however, no published case series describing this syndrome. Objectives: To characterize the cause and outcomes for horses referred for investigation of episodic collapse. Animals: Twenty‐five horses referred for investigation of single or multiple episodes of collapse. Methods: Retrospective study. Clinical records from the Dick Vet Equine Hospital, University of Edinburgh from November 1995 to July 2009 were searched using the following keywords: collapse, collapsing, fall, syncope. Collapse was defined as an incident in which the horse lost postural tone with or without progression to recumbency and with or without loss of consciousness. Long‐term follow‐up information was obtained by telephone conversation with the owner. Results: A final diagnosis was reached in 11 cases, namely cardiac arrhythmia (4), right‐sided heart failure (1), hypoglycemia (2), generalized seizures (2), and sleep disorder (2). A presumptive diagnosis was reached in 8 cases, namely neurocardiogenic syncope (5), exercise‐induced pulmonary hemorrhage (2), and generalized seizures (1). No diagnosis was reached in 6 cases despite comprehensive investigations. Three horses were euthanized at presentation. Treatment was attempted in 9 horses with 6 cases having successful outcome before discharge. Follow‐up information was available for 14 of 19 horses discharged from the hospital. Only 1 of these horses was observed to collapse after discharge. Conclusions and Clinical Importance: Definitive diagnosis was more likely to be reached in cases with multiple episodes of collapse. Horses in which 1 episode of collapse occurred did not necessarily collapse again.     

Cellular localization of Visudyne® as a function of time after local injection in an in vivo model of squamous cell carcinoma: an investigation into tumor cell death

Objective To evaluate the effects of time on cellular localization of Visudyne^® after local injection. Animals Twenty athymic nude mice. Procedures A squamous cell carcinoma (SCC) cell line (A‐431) was injected into right and left dorsolumbar subcutaneous tissue of each mouse, representing treatment (T) and control (C) tumors. In experiment 1 (Exp 1; n = 10) and 2 (Exp 2; n = 10), the T tumors received a local injection of Visudyne^® (0.1 mg/cm³), and C tumors received an equal dose of 5% dextrose in water (D5W). Mice were randomly subdivided into two groups (A and B; n = 5 per group). Mice in Exp 1A and B were sacrificed 1 and 30 min after local injection, respectively. Experiment 1A and B tumors were evaluated by fluorescence microscopy to determine drug localization. Experiment 2A and B tumors were exposed to LED illumination 1 and 30 min after injection, respectively, and evaluated by transmission electron microscopy (TEM) to determine ultrastructural tumor cell damage. Results Fluorescence was detected within the cytoplasm of T tumors in both Exp 1A and B. Significance was detected in fluorescence intensity between T1 min vs. T30 min (P = 0.03) and between T1 min and C1 min tumors (P = 0.01), respectively. Tumors in Exp 2A and B demonstrated evidence of apoptotic cell death. Conclusions Fluorescence microscopy demonstrated higher Visudyne^® concentration within SCC cytoplasm of 1 min compared with 30‐min tumors. Transmission electron microscopy results revealed that tumors treated by photodynamic therapy (PDT) within 30 min of local injection undergo cellular apoptosis.