Juxtacrine and paracrine interactions of rat marrow-derived mesenchymal stem cells, muscle-derived satellite cells, and neonatal cardiomyocytes with endothelial cells in angiogenesis dynamics.

Research into angiogenesis has contributed to progress in the fast-moving field of regenerative medicine. Designing coculture systems is deemed a helpful method to understand the dynamic interaction of various cells involved in the angiogenesis process. We investigated the juxtacrine and paracrine interaction between 3 different cells, namely rat marrow-derived mesenchymal stem cells (rMSCs), rat muscle-derived satellite cells (rSCs), and rat neonatal cardiomyocytes (rCMs), and endothelial cells (ECs) during angiogenesis process. In vitro Matrigel angiogenesis assay was performed whereby ECs were monocultured or cocultured with rMSCs, rSCs, and rCMs or their conditioned media (CM). In addition, in vivo Matrigel plug assay for angiogenesis was conducted to assess the angiogenic potential of the rCM-, rMSC-, and rSC-derived CM. Our results demonstrated that the rMSCs, rSCs, and rCMs elongated along the EC tubules, whereas the rMSCs formed tube-like structures with sprouting tip cells, leading to improved angiogenesis in the coculture system. Moreover, the rMSC- and rSC-derived CM significantly improved angiogenesis tube formation on Matrigel, accelerated EC chemotaxis, and increased the arteriolar density, vascularization index, and vascularization flow index in the Matrigel plug in vivo. Western blotting showed that rMSCs secreted a high level of vascular endothelial growth factor, basic fibroblast growth factor, and stromal-derived factor-1-alpha. Tie2 is also shed from rMSCs. This study demonstrated that stem cells interact with ECs in the juxtacrine and paracrine manner during angiogenesis, and marrow MSCs have superior angiogenic properties.

Bovine immune-mediated hemolytic anemia: 13 cases (November 2008-August 2009).

BACKGROUND: Immune-mediated hemolytic anemia (IMHA) occurs in cattle; however, there are few reported cases. OBJECTIVE: The aim of this study was to investigate the prevalence of IMHA in cattle with anemia, describe the associated clinical and laboratory findings, including osmotic fragility, and identify potential causative infectious agents or drugs. METHODS: This study included 42 anemic cattle (HCT < 27.5%) comprising 31 females and 11 bulls with a mean age of 3.5 years referred to the University of Tehran Veterinary Teaching Hospital during a 10-month period. CBCs, saline osmotic fragility tests, direct Coombs' tests, and biochemical profiles were performed, and blood smears were evaluated for spherocytosis, parasites, and microscopic agglutination. Five clinically healthy cattle were used as controls for testing osmotic fragility of RBCs. RESULTS: The Coombs' test was positive in 13/42 (30%) cattle; 5 had no evidence of concurrent disease or history of drug administration, and 8 had underlying or concurrent diseases, positivity for BLV, or exposure to drugs. The HCT (mean ± SE) of Coombs'-positive cattle (16 ± 1.7%) was significantly lower than that of Coombs'-negative animals (21 ± 0.8%). Hematologic and biochemical findings in cattle with IMHA included anisocytosis (2), polychromasia (2), basophilic stippling (2), spherocytosis (2), hyperfibrinogenemia (5), left-shifted neutrophilia (3), and hyperbilirubinemia (8). RBCs from Coombs'-positive anemic cattle were more fragile than those from Coombs'-negative anemic cattle. Four osmotically different populations of RBCs were detected in cattle with IMHA, whereas RBC populations were homogeneous in the Coombs'-negative anemic cattle and in normal cattle. CONCLUSION: IMHA was identified in a significant proportion of anemic cattle. Idiopathic IMHA and IMHA secondary to infectious diseases and administration of certain drugs occur in cattle.

Identification of different Theileria species (Theileria lestoquardi, Theileria ovis, and Theileria annulata) in naturally infected sheep using nested PCR-RFLP.

Ovine theileriosis is an important hemoprotozoal disease of sheep and goats in tropical and subtropical regions that leads to economic losses in these animals. A nested PCR-restriction fragment length polymorphism (RFLP) was carried out to identification Theileria species in sheep in some area in western half of Iran (Sari, Rasht, Urmia, Ilam, and Ahvaz). Two hundred and fifty blood samples were taken from sheep during tick activating season (summer of 2008). Microscopic examination revealed that 9.2% (23/250) sheep were infected by Theileria spp. piroplasms. Parasitemia ranged from 0.011% to 0.015%. In nested PCR assessment of DNA samples, 32.8% (82/250) sheep were positive. The negative samples were confirmed by amplifying of ovine beta-actin gene as an internal control. The differentiation of Theileria species was based on RFLP patterns using three restriction enzymes: HpaII, Rsa1, and Bsh 1285I. Out of 82 positive samples, 54.8% (45/82) and 40.2% (33/82) were positive for Theileria lestoquardi and Theileria ovis respectively. Mixed infection was detected in 4.8% (4/82) cases. Based on their PCR product digestion pattern with HpaII (1178, 900, 278, and 106 bp), it seemed to be mixture of Theileria annulata and T. lestoquardi. The presence of T. annulata was supported by sequence analysis. This is the first report of naturally infected sheep with T. annulata in Iran. Geographical distribution of Theileria species in sheep is shown according to the result of microscopy and nested PCR and RFLP data.