Megakaryocyte-stromal cell interactions: Effect on megakaryocyte proliferation, proplatelet production, and survival.

Bone marrow stromal cells provide a proper environment for the development of hematologic lineages. The incorporation of different stromal cells into in vitro culture systems would be an attractive model to study megakaryopoiesis and thrombopoiesis. Our objective was to evaluate the participation of different types of stromal cells in in vitro megakaryopoiesis, thrombopoiesis, and megakaryocyte (MK) survival. CD34-positive progenitors from umbilical cord blood were differentiated into MK precursors and then cocultured with umbilical vein endothelial cells (HUVECs), bone marrow mesenchymal stem cells (MSCs), skin fibroblasts (SFs) (all human), or the mouse fibroblast cell line L929. The number of MKs (CD61-positive cells) was increased in the presence of HUVECs and SFs, whereas L929 cells decreased total and mature MK counts. With respect to thrombopoiesis, HUVECs increased proplatelet (PP)-producing MKs, while MSCs, L929 cells, and SFs had the opposite effect (immunofluorescence staining and microscopic analysis). MK survival was enhanced in MSC and SF co-cultures, as assessed by evaluation of pyknotic nuclei. However, HUVECs and L929 did not prevent apoptosis of MKs. Reciprocally, the cloning efficiency of MSCs was decreased in the presence of MKs, while the ability of stromal cells (either MSCs or SFs) to produce the extracellular matrix proteins type III collagen, fibronectin, dermatan sulfate, heparan sulfate, and prolyl 4-hydroxylase subunit ß was preserved. These data indicate that each stromal cell type performs distinctive functions that differentially modulate MK growth and platelet production and, at the same time, that MKs also modify stromal cell behavior. Overall, our results highlight the relevance of considering the influence of stromal cells in MK research as well as the close interplay of different cell types within the bone marrow milieu.

Oligonucleotide IMT504 Improves Glucose Metabolism and Controls Immune Cell Mediators in Female Diabetic NOD Mice.

Type 1 diabetes occurs as a consequence of progressive autoimmune destruction of beta cells. A potential treatment for this disease should address the immune attack on beta cells and their preservation/regeneration. The objective of this study was to elucidate whether the immunomodulatory synthetic oligonucleotide IMT504 was able to ameliorate diabetes in NOD mice and to provide further understanding of its mechanism of action. We found that IMT504 restores glucose homeostasis in a diabetes mouse model similar to human type 1 diabetes, by regulating expression of immune modulatory factors and improving beta cell function. IMT504 treatment markedly improved fasting glycemia, insulinemia, and homeostatic model assessment of beta cell function (HOMA-Beta cell) index. Moreover, this treatment increased islet number and decreased apoptosis, insulitis, and CD45+ pancreas-infiltrating leukocytes. In a long-term treatment, we observed improvement of glucose metabolism up to 9 days after IMT504 cessation and increased survival after 15 days of the last IMT504 injection. We postulate that interleukin (IL)-12B (p40), possibly acting as a homodimer, and Galectin-3 (Gal-3) may function as mediators of this immunomodulatory action. Overall, these results validate the therapeutic activity of IMT504 as a promising drug for type 1 diabetes and suggest possible downstream mediators of its immunomodulatory effect.

Can QT/RR relationship differentiate between low- and high-risk patients with hypertrophic cardiomyopathy?

BACKGROUND: Abnormal dynamicity of repolarization is considered to be a marker of myocardial vulnerability contributing to increased risk of arrhythmic events and sudden death. However, little is known about QT dynamics in hypertrophic cardiomyopathy (HCM). In this study, we aimed to evaluate ventricular repolarization by QT dynamicity in patients with HCM, focusing on its value to define if it is able to differentiate among low- and high-risk HCM patients. METHODS: The linear regression slopes of the QT interval, measured to the apex and to the end of the T wave plotted against RR intervals (QTapex/RR and QTend/RR slopes, respectively) were calculated from 24-hour Holter recordings using a standard algorithm in 36 HCM patients and 64 control subjects. RESULTS: QTapex/RR and QTend/RR slopes were significantly steeper in the HCM patients in contrary to healthy control subjects (QTapex/RR = 0.22 + 0.08 vs 0.20 + 0.05, P = 0.0367; QTend/RR = 0.25 + 0.10 vs 0.20 + 0.06, P = 0.023). Moreover, the slopes of QTend/RR and QTapex/RR of high-risk patients were significantly steeper than those of control subjects while no significant differences were found among low-risk HCM patients and control subjects and only QTe/RR of high-risk patients was significantly different between low- and high-risk HCM patients. CONCLUSIONS: Our study results suggest that QT dynamicity is impaired in patients with HCM and may help to differentiate among low- and high-risk patients. Further studies are needed to elucidate the prognostic significance and clinical implications of impaired ventricular repolarization in patients with HCM.