ABSTRACT
Immune-mediated hemolytic anemia (IMHA) is the most common autoimmune disease in dogs. This study was conducted to evaluate prednisolone and azathioprine therapeutic protocols for the management of idiopathic IMHA in dogs. The anaemic dogs brought with clinical signs such as pale or icteric mucous membranes were screened for IMHA by saline agglutination and spherocyte count and confirmed by flow cytometry. The positive cases were further subjected to haematology, biochemistry, coagulation profile, MAT and polymerase chain reaction (PCR) for the diagnosis of underlying secondary causes like Babesia spp, Ehrlichia canis and Leptospira spp (secondary IMHA). Thirty two cases were positive for IMHA, out of which thirteen cases were primary (Idiopathic) IMHA (17.3 %) and remaining nineteen cases were secondary IMHA (82.7 %) due to underlying causes such as Babesia gibsoni (13), Ehrlichia canis (3) and Leptospira spp. (3) respectively. Immunosuppressive therapy with prednisolone and prednisolone in combination with azathioprine and specific therapy of etiological agent with supportive therapy was used. Significant increase in Hb, PCV, RBC and thrombocyte count, significant decrease in leucocyte, neutrophil, monocyte and total protein and significant increase in ALT activity was recorded after therapy. Prednisolone was found to be effective in the management of canine IMHA than prednisolone combined with azathioprine
ABSTRACT
Immune-mediated hemolytic anemia (IMHA) is the most common autoimmune disease in dogs. This study was conducted to study the incidence and clinicopathological changes of secondary IMHA, which is having any secondary underlying triggering causes. The anaemic dogs brought with clinical signs such as pale or icteric mucous membranes were screened for IMHA by saline agglutination and spherocyte count and confirmed by flow cytometry. The positive cases were further subjected to haematology, biochemistry, coagulation profile, MAT and polymerase chain reaction (PCR) for the diagnosis of underlying secondary causes like Babesia spp, Ehrlichia canis and Leptospira spp. (secondary IMHA). Thirty two cases were positive for IMHA, out of which thirteen cases were primary IMHA (17.3 %) and remaining nineteen cases were secondary IMHA (82.7 %) due to underlying causes such as Babesia gibsoni (13), Ehrlichia canis (3) and Leptospira spp. (3) respectively. Highest incidence was observed in Labrador dogs with age group of 2-8 years in male dogs. The most common clinicopathological findings were anaemia with reduced Hb, PCV and RBC, leucocytosis, neutrophilia, monocytosis, thrombocytopenia, elevated PT, BUN, Creatinine, ALT, ALP, hypoalbuminemia and hyperbilirubinaemia
ABSTRACT
Background & objectives: Mesencymal stem cells (MSCs) derived from foetal tissues present a multipotent progenitor cell source for application in tissue engineering and regenerative medicine. The present study was carried out to derive foetal mesenchymal stem cells from ovine source and analyze their differentiation to osteogenic linage to serve as an animal model to predict human applications. Methods: Isolation and culture of sheep foetal bone marrow cells were done and uniform clonally derived MSC population was collected. The cells were characterized using cytochemical, immunophenotyping, biochemical and molecular analyses. The cells with defined characteristics were differentiated into osteogenic lineages and analysis for differentiated cell types was done. The cells were analyzed for cell surface marker expression and the gene expression in undifferentiated and differentiated osteoblast was checked by reverse transcriptase PCR (RT PCR) analysis and confirmed by sequencing using genetic analyzer. Results: Ovine foetal samples were processed to obtain mononuclear (MNC) cells which on culture showed spindle morphology, a characteristic oval body with the flattened ends. MSC population CD45-/CD14- was cultured by limiting dilution to arrive at uniform spindle morphology cells and colony forming units. The cells were shown to be positive for surface markers such as CD44, CD54, integrinβ1, and intracellular collagen type I/III and fibronectin. The osteogenically induced MSCs were analyzed for alkaline phosphatase (ALP) activity and mineral deposition. The undifferentiated MSCs expressed RAB3B, candidate marker for stemness in MSCs. The osteogenically induced and uninduced MSCs expressed collagen type I and MMP13 gene in osteogenic induced cells. Interpretation & conclusions: The protocol for isolation of ovine foetal bone marrow derived MSCs was simple to perform, and the cultural method of obtaining pure spindle morphology cells was established. Criteria proposed for defining MSCs by this study includes the cell adherence to culture plates, specific surface protein profiles and differentiation to osteogenic lineage. The MSCs and osteogenic differentiated cells in this ovine animal model may serve as a large source for stem cell applications in regenerative medical therapies.