Rapid,effective and user-friendly immunophenotyping of canine lymphoma using a personal flow cytometer     

Stratos Papakonstantinou  Inese Berzina  Amanda Lawlor  Emma J O’Neill  Peter J O’Brien 《Irish veterinary journal》2013,66(1):6

Background

Widespread use of flow cytometry for immunophenotyping in clinical veterinary medicine is limited by cost and requirement for considerable laboratory space, staff time, and expertise. The Guava EasyCyte Plus (Guava Technologies, Hayward, CA, US) is the first, personal, bench-top flow cytometer designed to address these limitations.

Objective

The aim of this study was to adapt the immunohistochemical protocol used for immunophenotyping of canine lymphoma to the personal flow cytometer for rapid, effective and user-friendly application to the diagnosis and prognosis of canine lymphoma and to demonstrate its practicality for widespread veterinary application. Performance of the personal flow cytometer for immunophenotyping T and B lymphocytes in blood and lymph nodes from normal dogs and dogs with lymphoproliferative disease, was assessed using only two monoclonal antibodies (against CD3 and CD21), and by comparison with analysis using two conventional flow cytometers.

Methods

26 dogs with lymphoproliferative disease (23 with lymphoma, 3 with lymphocytic leukaemia) were studied along with 15 controls (2 non-lymphoma lymph nodes and 13 non-leukemic bloods. Lymphocytes were immunostained with fluorescent-labeled, monoclonal antibodies against CD3 and CD21. To assess the effectiveness of the personal flow cytometer in discrimination between T and B cell immunophenotypes, T and B cell counts for half the samples (14 blood and 11 lymph node) were also determined using the same method and conventional flow cytometers (FACSCalibur, Cyan Dako). To assess the effectiveness of the personal flow cytometer in discriminating between leukocyte types, lymphocyte differential counts were determined for 21 blood samples and compared with those from automated hematology analyzers (CELL-DYN 3500, n=11 and ADVIA 2120, n=10). Quality and sub-cellular distribution of immunostaining was assessed using fluorescence microscopy.

Results

The protocol for immunophenotyping took 2 to 3 hours to complete from the point of receipt of sample to reporting of immunophenotype. The personal flow cytometer differential lymphocyte counts correlated highly (n=20; r=0.97, p<0.0001) with those of automated haematology analyzers. The personal flow cytometer counts consistently, but mildly, underestimated the percentages of lymphocytes in the samples (mean bias of -5.3%.). The personal flow cytometer immunophenotype counts were indistinguishable from those of conventional flow cytometers for both peripheral blood samples (n=13; r=0.95; p<0.0001; bias of -1.1%) and lymph node aspirates (n=11,r=0.98; p<0.001; bias of 1%). All but one leukemic and one lymphomatous lymph node sample, out of 26 samples of dogs with lymphoproliferative disease analyzed, could be immunophenotyped as either B or T cells.

Conclusions

We conclude that use of only 2 monoclonal antibodies is sufficient for immunophenotyping most cases of canine lymphoma by flow cytometry and enables rapid immunophenotyping. The personal flow cytometer may be as effectively used for immunophenotyping canine lymphoma as conventional flow cytometers. However, the personal flow cytometer is more accessible and user-friendly, and requires lower sample volumes.  相似文献   

The use of lung biopsy to determine early lung pathology and its association with health and production outcomes in feedlot steers     

Brandy A. Burgess  Steve H. Hendrick  Colleen M. Pollock  Sherry J. Hannon  Sameeh M. Abutarbush  Amanda Vogstad  G. Kee Jim  Calvin W. Booker 《Canadian journal of veterinary research》2013,77(4):281-287

The objectives of this study were to determine if percutaneous lung biopsy can be used to characterize early pathologic changes in bovine lung associated with bovine respiratory disease (BRD), to determine if specific infectious respiratory pathogens can be identified in association with these changes, and to determine whether pulmonary pathology at arrival and at the time of initial diagnosis are associated with health and production outcomes. One hundred auction-market derived crossbred steer calves from a commercial feedlot in southern Alberta were included in this study. A percutaneous lung biopsy technique was used to obtain lung samples from the right middle lung. Steers were sampled 295 times yielding 283 samples with 210 (74%) containing lung tissue. Overall, histopathological changes were observed in 20 (9.5%) of lung biopsy samples. There were too few samples with pathology to reveal an association between lung pathology and subsequent health events. In general, percutaneous lung biopsy can be done safely on feedlot steers in a commercial feedlot setting with few clinical side effects. This technique did not prove useful as a diagnostic tool or prognostic indicator for early BRD. However, it may be useful for the diagnosis of BRD in targeted populations of commercial feedlot steers.  相似文献   

Hypoperfusion and acute traumatic coagulopathy in severely traumatized canine patients     

Amanda L. Abelson DVM  DACVECC  Therese E. O'Toole DVM  DACVIM  DACVECC  Andrea Johnston DVM  Meghan Respess DVM  Armelle M. de Laforcade DVM  DACVECC 《Journal of Veterinary Emergency and Critical Care》2013,23(4):395-401

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Letter to the Editor     

Amanda Boag MA  VetMB  DACVIM  DACVECC 《Journal of Veterinary Emergency and Critical Care》2013,23(4):376-376

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The use and interpretation of serum retinol distributions in evaluating the public health impact of vitamin A programmes     

Palmer AC  West KP  Dalmiya N  Schultink W 《Public health nutrition》2012,15(7):1201-1215

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Formation of conidial anastomosis tubes and melanization of appressoria are antagonistic processes in Colletotrichum spp. from apple     

Amanda Emy Gonçalves  Aline Cristina Velho  Marciel J. Stadnik 《European journal of plant pathology / European Foundation for Plant Pathology》2016,144(3):497-507

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Anadenanthera colubrina (Vell.) Brenan produces steroidal substances that are active against Alternaria alternata (Fr.) Keissler and that may bind to oxysterol‐binding proteins     

Viviane AC Campos  Fabiano J Perina  Eduardo Alves  Jaqueline Sartorelli  Amanda M Moura  Denilson F Oliveira 《Pest management science》2014,70(12):1815-1822

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Characterization of catalytic efficiency parameters of brain cholinesterases in tropical fish     

Caio Rodrigo Dias de Assis  Amanda Guedes Linhares  Vagne Melo Oliveira  Renata Cristina Penha França  Juliana Ferreira Santos  Marina Marcuschi  Elba Verônica Matoso Maciel Carvalho  Ranilson Souza Bezerra  Luiz Bezerra Carvalho Jr. 《Fish physiology and biochemistry》2014,40(6):1659-1668

Brain cholinesterases from four fish (Arapaima gigas, Colossoma macropomum, Rachycentron canadum and Oreochromis niloticus) were characterized using specific substrates and selective inhibitors. Parameters of catalytic efficiency such as activation energy (AE), k _cat and k _cat/k _m as well as rate enhancements produced by these enzymes were estimated by a method using crude extracts described here. Despite the BChE-like activity, specific substrate kinetic analysis pointed to the existence of only acetylcholinesterase (AChE) in brain of the species studied. Selective inhibition suggests that C. macropomum brain AChE presents atypical activity regarding its behavior in the presence of selective inhibitors. AE data showed that the enzymes increased the rate of reactions up to 10¹² in relation to the uncatalyzed reactions. Zymograms showed the presence of AChE isoforms with molecular weights ranging from 202 to 299 kDa. Values of k _cat and k _cat/k _m were similar to those found in the literature.  相似文献   

Clopidogrel induced suppression of bovine platelet activation in vitro and a preliminary study of its effect on the development of Mannheimia haemolytica induced pneumonia     

Coomber BL  Mitchell GB  Starr AE  Minhas K  Tamblyn A  Shewen PE  Gentry PA 《Veterinary journal (London, England : 1997)》2006,171(1):126-134

We report here on the influence of the platelet antagonist clopidogrel (Plavix) on bovine platelet function. We first evaluated the capacity of clopidogrel to inhibit adenosine diphosphate (ADP)-stimulated platelet function in the bovine species, using an ex vivo approach with blood from treated animals. Platelets isolated from treated calves displayed rapid and consistent reduction in function (aggregation, thromboxane production) upon ADP, but not platelet activating factor (PAF), stimulation. We then examined the possibility that clopidogrel could influence Mannheimia haemolytica pneumonia pathobiology using an experimental challenge model. We were unable to detect significant differences between clopidogrel treated and untreated animals when challenged with intra-tracheal inoculation of M. haemolytica. There was a trend towards inhibition of platelet degranulation in the affected regions of lungs from clopidogrel treated calves, and pre-treated challenged animals had similar amounts of fibrin deposition and enhanced fibrous tissue formation in their lungs when compared with control counterparts.  相似文献   

Adjuvant-enhanced antibody responses in the absence of toll-like receptor signaling     

Gavin AL  Hoebe K  Duong B  Ota T  Martin C  Beutler B  Nemazee D 《Science (New York, N.Y.)》2006,314(5807):1936-1938

Innate immune signals mediated by Toll-like receptors (TLRs) have been thought to contribute considerably to the antibody-enhancing effects of vaccine adjuvants. However, we report here that mice deficient in the critical signaling components for TLR mount robust antibody responses to T cell-dependent antigen given in four typical adjuvants: alum, Freund's complete adjuvant, Freund's incomplete adjuvant, and monophosphoryl-lipid A/trehalose dicorynomycolate adjuvant. We conclude that TLR signaling does not account for the action of classical adjuvants and does not fully explain the action of a strong adjuvant containing a TLR ligand. This may have important implications in the use and development of vaccine adjuvants.  相似文献