VCAM1 Human ELISA Kit

Burn injury has been shown to significantly dampen erythropoiesis in both burn patients and in murine models. Our previous findings elucidated the erythropoietin independent defects in red cell development stages involving erythroid progenitor production and late stage erythroblast enucleation processes. We hypothesized that macrophages (Mo) in erythroblast islands (EBI) could be yet another roadblock impeding erythropoiesis following burn injury. Here we highlight that the methodology to study EBI can be achieved with single cell suspensions using a simple technique such as flow cytometry, as obtaining the central erythroblast island macrophages (EBIMos) of interest is a delicate process. We elucidated the requisite of EBIMo from the erythroblast as well as the Mo perspective. In addition to the primary erythropoiesis organ, the bone marrow (BM), spleens were also examined for extra-medullary erythropoiesis. Femurs and spleens were harvested from adult mice (B6D2F1) subjected to 15% total body surface area (TBSA) scald burn (B) or sham burn (S). Total bone marrow cells (TBM) and splenocytes were probed for total erythrons, early and late erythroblasts and EBIMo by flow cytometry. There was only a marginal increase in the number of EBIMo after burn, but, between the signatures of EBIMo, Siglec-1 expression (MFI) was reduced by 40% in B with and a parallel 44% decrease in TBM erythrons in the BM. There were more (2.5-fold) EEBs and less LEBs (2.4-fold) per million TBM cells in B; with a corresponding decrease in Siglec-1 and Ly6G expressions in EBIMo associated with EEB. Conversely, extra-medullary erythropoiesis was robust in spleens from B. Not only were the numbers of EBIMos increased in B (p < 0.002), both EEBs and LEBs associated with EBIMo were higher by 30 and 75%, respectively. Importantly, an increase in Siglec-1 and Vcam1 expressing F480(+) splenic macrophages was observed after burn injury. Therefore, stagnant EEBs in the BM after burn injury could be due to low Siglec1 expressing EBIMo, which perhaps impede their maturation into LEBs and reticulocytes. Repercussion of myeloid cell phenotype specific to BM after burn injury could plausibly account for a defective late stage RBC maturation resulting in anemia of critical illness. Summary Sentence: Characterization of erythroblast island macrophages (EBIMo) in the bone marrow and spleen at different stages of erythropoiesis after burn injury.

Adipose tissues are related to the meat quality and economic value of cattle. In particular, the extracellular matrix (ECM) is an important factor that influences adipose tissue remodeling. The objective of this study was to measure the expression levels of ECM-related genes in different adipose tissues from bulls, cows, and steers of Korean cattle (Hanwoo). The sample tissues were separated from omental (OM), subcutaneous (SC), and intramuscular (IM) adipose tissues. The expression levels of 24 ECM-related genes and proteins were determined using real-time PCR and Western blot analysis, respectively. We also observed that castration resulted in increased (P < 0.05) expression of 17 genes, whereas it caused decreased (P < 0.05) expression of COL1A2, MMP1, MMP3, MMP7, and VCAM1 in the OM adipose tissue of cows and steers compared to bulls. The mRNA levels of 24 ECM-related genes were similar between IM and SC adipose tissues. Furthermore, except for ITGB1 (IM), ICAM1 (IM), and COL1A1 (OM), the expression levels were generally similar between cows and steers compared to bulls. The FM, MMP7, and LOX mRNA levels exhibited significantly strong (P < 0.001) positive correlations (r=0.74, r=0.73, and r=0.74, respectively) with the marbling score. The protein levels can reflect the mRNA levels. Therefore, our findings suggest that castration affects the expression levels of ECM-related genes, which is in accordance with different fat depots.