Role of tumour necrosis factor receptor-1 and nuclear factor-κB in production of TNF-α-induced pro-inflammatory microparticles in endothelial cells.

Tumour necrosis factor-α (TNF-α) is upregulated in many inflammatory diseases and is also a potent agent for microparticle (MP) generation. Here, we describe an essential role of TNF-α in the production of endothelial cell-derived microparticles (EMPs) in vivo and the function of TNF-α-induced EMPs in endothelial cells. We found that TNF-α rapidly increased blood levels of EMPs in mice. Treatment of human umbilical vein endothelial cells (HUVECs) with TNF-α also induced EMP formation in a time-dependent manner. Silencing of TNF receptor (TNFR)-1 or inhibition of the nuclear factor-κB (NF-κB) in HUVECs impaired the production of TNF-α-induced EMP. Incubation of HUVECs with PKH-67-stained EMPs showed that endothelial cells readily engulfed EMPs, and the engulfed TNF-α-induced EMPs promoted the expression of pro-apoptotic molecules and upregulated intercellular adhesion molecule-1 level on the cell surface, which led to monocyte adhesion. Collectively, our findings indicate that the generation of TNF-α-induced EMPs was mediated by TNFR1 or NF-κB and that EMPs can contribute to apoptosis and inflammation of endothelial cells.

Tie2-mediated multidrug resistance in malignant gliomas is associated with upregulation of ABC transporters.

Resistance and relapse are still primary causes that result in poor effectiveness of chemotherapy in malignant gliomas. Therefore, development of new therapeutic strategies requires the identification of key molecular pathways regulating chemoresistance. We previously found that abnormal high expression of the Tie2 receptor in gliomas was associated with tumor malignancy. Here, we studied the role of Tie2 activation in drug resistance by testing the cytotoxicity of several chemotherapeutic drugs in a panel of human glioma cell lines and brain tumor stem cells and found that Tie2 activation was significantly related to chemoresistance. The essential role of Tie2 in this phenotype was illustrated by silencing Tie2 using specific siRNA, and the subsequent abrogation of the angiopoietin 1 (Ang1)-mediated chemoresistance. Using quantitative real-time PCR and functional drug efflux studies, we observed that Tie2 activation resulted in increased expression of ATP-binding cassette (ABC) transporters. Consistent with these results, downmodulation of ABCG2 or ABCC2 resulted in the inability of Tie2 activation to induce a chemoresistant phenotype. Our results indicate that Tie2 activation may be important in modifying the evolution of gliomas during conventional chemotherapy regimens, and open new avenues for the search of more effective therapies to avoid the inevitable brain tumor recurrence.

Radical scavenging-linked antioxidant activity of ethanolic extracts of diverse types of extra virgin olive oils.

The present study evaluated the radical scavenging-linked antioxidant activity of hexane/80% ethanol extracts from several types of extra virgin olive oils (EVOOs) derived from varieties arbequina, hojiblanca, picual, their blends, and pure olive oil (POO). The antioxidant potential of the olive oil extracts was assessed by radical scavenging assays using DPPH (2, 2-diphenyl-1-picrylhydrazyl), ABTS (2, 2'-azino-bis (3-ethylbenzthiazoline-6-sulfonic acid), and hydroxyl radical, as well as hydrogen peroxide and superoxide anion inhibitory activities. Electron donating ability (EDA) using DPPH assay of 80% ethanol extracts from EVOOs, except arbequina oil, was significantly higher than POO. EDA was markedly higher in blended and picual EVOOs than the extracts from arbequina and hojiblanca EVOOs (P < 0.05). Similarly, ABTS radical scavenging activity of the extracts from the EVOOs was in order of picual EVOO > blended EVOO > hojiblanca EVOO >or= POO >or= arbequina EVOO. Further, the superoxide anion scavenging activity of blended, picual, and arbequina EVOOs was significantly higher than that of hojiblanca EVOO and POO, which were barely detectable. Hydroxyl radical scavenging activity of arbequina and hojiblanca was higher than that of blended, picual EVOOs, and POO. In addition, hydrogen peroxide scavenging activity of the extracts from blended, arbequina, hojiblanca, picual EVOOs, and POO was 63.1 +/- 3.1%, 44.4 +/- 10.2%, 52.0 +/- 2.7%, 71.8 +/- 2.5%, and 35.7 +/- 10.0%, respectively. Our results indicate that ethanol extracts of several EVOOs contained higher radical scavenging and antioxidant activity than the POO. This antioxidant potential is partly due to the phenolic compounds present in different olive oil grade and is influenced by cultivar type.

Angiopoietin-2 decreases vascular endothelial growth factor expression by modulating HIF-1 alpha levels in gliomas.

Angiogenesis is thought to depend on a perfectly coordinated balance between endogenous-positive and negative regulatory factors. Of these factors, the vascular endothelial growth factor (VEGF) and angiopoietins (Angs) seem to play an essential role. Recently, we reported the expression of the Ang-natural receptor, Tie2, in neoplastic astrocytic cells within gliomas. Because of the VEGF/Ang2 functional partnership together with the presence of Tie2 in gliomas, we hypothesized a role of Ang2 on the modulation of VEGF levels in these tumors. We examined the effect of Ang2 on VEGF expression in a panel of glioma cells, which showed that Ang2 inhibited VEGF expression at both mRNA and protein levels in Tie2-expressing cells, but not in Tie2-negative cells. VEGF promoter analysis showed that Ang2 regulated VEGF expression at the transcriptional level in relation to a decrease in HIF-1alpha expression and HIF-DNA-binding activity. Tie2 silencing by siRNA rescued the Ang2-mediated downmodulation of VEGF, suggesting an essential role for Tie2 in this regulatory loop. To our knowledge, this is the first report on the role of Ang2/Tie2 in the regulation of HIF-1alpha/VEGF expression, providing additional evidence of the intrinsic coordination that occurs among these factors during angiogenesis.

Development of a measure of resident satisfaction with the nursing home.

A satisfaction instrument specifically designed for use with nursing home residents, the Satisfaction with the Nursing Home Instrument (SNHI), was developed and tested with a sample of 110 nursing home residents from three proprietary facilities in Minnesota. As hypothesized, significant relationships were found between SNHI scores and measures of affect (negatively associated with depression and positively associated with morale), providing support for the construct validity of the scale. The lack of a significant relationship between SNHI scores and both age and mental status confirmed the predicted divergent validity of the instrument. The alpha coefficient for the 29-item scale was 0.81.