Evidence for proangiogenic cellular and humoral systemic response in patients with acute onset of spinal cord injury.

CONTEXT/OBJECTIVE: Traumatic spinal cord injury (SCI) leads to disruption of local vasculature inducing secondary damage of neural tissue. Circulating endothelial progenitor cells (EPCs) play an important role in post-injury regeneration of vasculature, whereas endothelial cells (ECs) reflect endothelial damage. METHODS: Twenty patients with SCI were assessed during the first 24 hours, at day 3, and day 7 post-injury and compared to 25 healthy subjects. We herein investigated EPC and EC counts by flow cytometry as well as the levels of soluble factors (SDF-1, HGF, VEGF, Ang2, EGF, endoglin, PLGF, FGF-2, ET-1, BDNF, IGF-1) regulating their migration and proangiogenic function. To better characterize peripheral blood (PB) cells, global gene expression profiles of PB-derived cells were determined using genome-wide RNA microarray technology. RESULTS: We found significantly higher EPC (CD34(+)/CD133(+)/VEGFR2(+)) as well as EC (VEGFR2(+)) count in PB of patients with SCI within 7 days post-injury and the increased HGF, ET-1, Ang2, EGF, and PLGF plasma levels. Global gene expression analysis revealed considerably lower expression of genes associated with both innate and adaptive immune response in PB cells in patients. CONCLUSION: Collectively, our findings demonstrate that SCI triggers bone marrow-derived EPC mobilization accompanied by increased circulating EC numbers. Significant changes in both chemoattractive and proangiogenic cytokines plasma levels occurring rapidly after SCI suggest their role in SCI-related regenerative responses to injury. Broadened knowledge concerning the mechanisms governing of human organism response to the SCI might be helpful in developing effective therapeutic strategies.

Sodium iodate selectively injuries the posterior pole of the retina in a dose-dependent manner: morphological and electrophysiological study.

Sequential morphological and functional features of retinal damage in mice exposed to different doses (40 vs. 20 mg/kg) of sodium iodate (NaIO(3)) were analyzed. Retinal morphology, apoptosis (TUNEL assay), and function (electroretinography; ERG) were examined at several time points after NaIO(3) administration. The higher dose of NaIO(3) caused progressive degeneration of the whole retinal area and total suppression of scotopic and photopic ERG. In contrast, the lower dose induced much less severe degeneration in peripheral part of retina along with a moderate decline of b- and a-wave amplitudes in ERG, corroborating the presence of regions within retina that retain their function. The peak of photoreceptor apoptosis was found on the 3rd day, but the lower dose induced more intense reaction within the central retina than in its peripheral region. In conclusion, these results indicate that peripheral area of the retina reveals better resistance to NaIO(3) injury than its central part.

Morphology of the bone marrow, spleen and liver during hematopoietic cell mobilization with cyclophosphamide in mice.

Cyclophosphamide (CY), the agent with cytoreductive activity, is widely exploited in cancer chemotherapy, and can be used alone or in combination with various cytokines and growth factors to stimulate the egress of hematopoietic stem/progenitor cells (HSPC) from the BM compartment. The aim of the present study was to exam the morphology and ultrastructure of the bone marrow, spleen and liver of mice injected intraperitoneally with a single dose of cyclophosphamide (200 mg/kg bw) and the localization of cells expressing markers of early hematopoietic cells in studied organs and the peripheral blood. We observed that the CY-induced morphological changes in the BM and spleen were reconstructed on day 4. of experiment, and the spleen was repopulated by HSPC on the 6th day. In this time, the highest number of c-Kit-R-positive cells was determined by flow cytometry in the peripheral blood. The results confirmed, that the egress of HSPC from the bone marrow into the peripheral blood was delayed compared to mice treated with G-CSF or GCS-F plus CY.

The influence of STAT5 antisense oligodeoxynucleotides on the proliferation and apoptosis of selected human cutaneous T-cell lymphoma cell lines.

STAT5 (signal transducers and activators of transcription) are suggested to play a role in the pathogenesis of leukaemia and lymphoma; however, their influence on the growth of cutaneous T-cell lymphoma cells is not clear enough. The aim of our study was to analyse the function of STAT5 proteins in the proliferation and apoptosis of selected cutaneous T-cell lymphoma cell lines (HUT 78; PB-1; HUT 102B), using antisense oligodeoxynucleotide (ODN) strategy. RT-PCR and Western blot were applied to analyse the expression of STAT5 after incubation with antisense ODN (AS ODN). The effect of ODN pretreatment on the cell clonogenecity was analysed in methylcellulose cultures. The process of apoptosis was estimated using two different flow cytometry (FACScan) methods: (1) combined Annexin V/propidium iodide staining, (2) the TUNEL method. Perturbation of STAT5 expression reduced the proliferation of the PB-1 cells after a 24-h exposure to antisense ODNs. Prolonged exposure (72 h) decreased the growth of each examined cell line, especially after antisense STAT5A (AS STAT5A) treatment. Incubation with AS STAT5 induced apoptosis in the population of HUT 78 and PB-1 cells. STAT5s may play a significant role in the growth and the process of apoptosis of selected human cutaneous T-cell lymphoma cells.