text:
Florian P. Limbourg1, Zhihong Huang2, Jean-Christophe Plumier2, Tommaso Simoncini3, Masayuki Fujioka2, Jan Tuckermann4, Günther Schütz4, Michael A. Moskowitz2 and James K. Liao1
1Cardiovascular Division, Brigham and Women's Hospital and Harvard Medical School, Boston, Massachusetts, USA2
Laboratory of Stroke and Neurovascular Regulation, Massachusetts General Hospital and Harvard Medical School, Boston, Massachusetts, USA3
Department of Reproductive Medicine and Child Development, University of Pisa, Pisa, Italy4
Department of Molecular Biology of the Cell I, German Cancer Research Center, Heidelberg, Germany
Address correspondence to: James K. Liao, Vascular Medicine Research, Brigham and Women's Hospital, 65 Landsdowne Street, Room 275, Cambridge, Massachusetts 02139, USA. Phone: (617) 768-8424; Fax: (617) 768-8425; E-mail: firstname.lastname@example.com.
Published December 1, 2002Submitted: March 19,
2002; Accepted: October 8,
Many cellular responses to corticosteroids involve the transcriptional modulation of target genes by the glucocorticoid receptor (GR). A rapid, non-nuclear effect of GR was found to mediate neuroprotection. High-dose corticosteroids (20 mg/kg intraperitoneally), given within 2 hours of transient cerebral ischemia, acutely increased endothelial nitric oxide synthase (eNOS) activity, augmented regional cerebral blood flow (CBF) by 40% to 50%, and reduced cerebral infarct size by 32%. These neuroprotective effects of corticosteroids were abolished by the GR antagonist RU486 and by inhibition of phosphatidylinositol 3-kinase (PI3K), and were absent in eNOS–/– mice. To determine the mechanism by which GR activated eNOS, we measured the effect of corticosteroids on PI3K and the protein kinase Akt. In a ligand-dependent manner, GR activated PI3K and Akt in vitro and in vivo caused NO-dependent vasodilation, which was blocked by cotreatment with RU486 or the PI3K inhibitor LY294002 but not by transcriptional inhibitors. Indeed, a mutant GR, which cannot dimerize and bind to DNA, still activated PI3K and Akt in response to corticosteroids. These findings indicate that non-nuclear GR rapidly activates eNOS through the PI3K/Akt pathway and suggest that this mechanism mediates the acute neuroprotective effects of corticosteroids through augmentation of CBF.
Steroid hormones are essential for normal development and stress responses. Corticosteroids bind to and activate the cytoplasmic glucocorticoid receptor (GR), a widely expressed steroid hormone receptor (1). In the classic model of steroid hormone action, GR acts as a ligand-dependent transcription factor by either activating or repressing gene expression through direct interactions with DNA or other transcription factors (2). The actions of GR are essential for survival. Mice lacking GR (GR–/–) die at birth (3); in contrast, overexpression of GR renders mice resistant to stress and endotoxic shock (4). Although the absence of GR is incompatible with life, DNA binding and transactivation of target genes by GR is not essential for development or survival, since mice with a targeted mutation in the dimerization domain of GR (GRdim), which impairs cooperative DNA binding and transactivation by GR homodimers, lack GR-dependent gene expression but survive to adulthood (5).
Some effects of corticosteroids on the brain and vascular system are very rapid, which makes transcriptional mechanisms of action unlikely. For example, behavioral changes in amphibians and increases of inositol trisphosphate in vascular smooth-muscle cells occur within minutes of corticosteroid administration (6, 7). Furthermore, the acute, antianaphylactic effects of high-dose corticosteroids in mice are unaltered by the transcriptional inhibitor actinomycin D (8). Thus, GR may exert important nontranscriptional functions. There is growing evidence that nontranscriptional actions of steroid hormone receptors regulate physiologically important processes. For example, osteocyte apoptosis is regulated by the androgen and estrogen steroid receptors through nontranscriptional activation of Src and mitogen-activated protein kinase (MAPK) (9). Vascular nitric oxide (NO) production and vasodilation by estrogen depend on nontranscriptional activation of endothelial NO synthase (eNOS) and are mediated by estrogen receptor α–dependent (ERα)-dependent activation of the phosphatidylinositol 3-kinase (PI3K)/Akt pathway (10–12). It is important to note that the vascular protective effects of estrogen are dependent on the nontranscriptional activation of eNOS through the PI3K/Akt pathway (11).
Endothelial-derived NO is a critical mediator of vascular integrity. Vascular NO production regulates cerebrovascular perfusion and protects against stroke by increasing collateral flow to the ischemic area (13). Mice lacking eNOS exhibit larger cerebral infarctions (14), and further inhibition of NOS activity by Nω-nitro-L-arginine methyl ester (L-NAME) decreases cerebral blood flow (CBF) and increases infarct size after ischemia in mice lacking neuronal nitric oxide synthase (nNOS) (15). In contrast, enhanced NO production by administration of the eNOS substrate, L-arginine, or upregulation of eNOS by statins confer protection from stroke (16, 17). Therefore, conditions that enhance eNOS activity could have beneficial effects on cerebrovascular disease.
Corticosteroids have been used in the treatment of stroke with conflicting results (18). Some clinical and experimental data, however, suggest a beneficial role for these steroids in reducing ischemic tissue injury. Corticosteroids reduce ischemic injury in myocardial infarction (19) and substantially reduce stroke size in focal cerebral ischemia (20–22). Indeed, we have recently shown that the anti-inflammatory and myocardial protective effects of acute steroid therapy are mediated by nontranscriptional activation of eNOS through the PI3K/Akt pathway (23). However, long-term corticosteroid therapy may be associated with an increased risk of stroke, due in part to transcriptional downregulation of eNOS expression and an increase in systemic blood pressure (24). The mechanism of stroke protection by steroids, therefore, is essentially unknown. Here, we examine whether non-nuclear signaling by GR could contribute to neuroprotection by a novel mechanism involving an increase in CBF.
Cells and reagents. Human endothelial cells were maintained in EGM-2 growth medium (Clonetics, San Diego, California, USA) and used at the third passage. Bovine aortic endothelial cells (BAECs) and COS7 cells were grown in DMEM (Life Technologies Gibco BRL, Grand Island, New York, USA) with 10% FCS. For experiments, cells were cultured for 48 hours in phenol red–free medium with charcoal-stripped FCS, and this procedure was followed by serum-starvation. Cells were incubated with dexamethasone (Sigma-Aldrich, St. Louis, Missouri, USA), RU486 (Biomol, Plymouth Meeting, Pennsylvania, USA), actinomycin D, LY294002 (Calbiochem, San Diego, California), or vehicle (DMSO). Phosphatidylinositol 4,5-bisphosphate was purchased from Biomol and [32P]γATP from New England Nuclear Life Science Products (Boston, Massachusetts, USA). The following antibodies were used: GR (rabbit, Santa Cruz Biotechnology, Santa Cruz, California, USA; mouse, Transduction Laboratories, Lexington, Kentucky, USA); MR (rabbit and goat, Santa Cruz Biotech); ERα (rabbit, Santa Cruz Biotech; mouse, Neomarkers, Freemont, California, USA); p85α (rabbit, Upstate Biotechnology, Lake Placid, New York, USA; mouse, Transduction Laboratories); P-Akt and Akt (rabbit, Cell Signaling Technology, Beverly, Massachusetts, USA); P-Tyr 4G10 (mouse, Upstate Biotechnology); eNOS (mouse, Transduction Labs); actin (rabbit, Sigma-Aldrich). All other reagents were purchased from Sigma-Aldrich.
PI3K assay. Approximately 500 μg of protein in buffer A (1% NP-40, 20 mM Tris [pH 7.4], 137 mM NaCl, 1 mM CaCl2, 1 mM MgCl2, 0.1 mM Na3VO4) were incubated with 1 μg of GR antibody and immunoprecipitated with 40 μl of 1:1 protein A/agarose. PI3K activity in the immunoprecipitate was assayed as described (25). The phospholipids were extracted with 160 μl of chloroform/methanol (1:1, v/v) and separated by borate thin-layer chromatography.
Akt kinase assay and Western blotting. Cells or tissues were homogenized in lysis buffer (1% Triton, 20 mM Tris [pH 7.4], 150 mM NaCl, 1 mM EDTA, 1 mM EGTA, 2.5 mM sodium pyrophosphate, 1 mM β-glycerolphosphate, 1 mM Na3VO4), and Akt kinase activity was determined by detection of phosphorylated GSK-3 fusion protein using the Akt kinase assay kit (Cell Signaling Technology) according to the manufacturer's guidelines. For Western blotting of cell extracts, typically 25–40 μg of protein were separated by SDS-PAGE, transferred to nitrocellulose membranes (Osmonics, Westborough, Massachusetts, USA), and probed with antibodies according to the manufacturer's instructions.
Transient transfection and reporter assays. BAECs were transfected for 6 hours with 8 μg of pGRE-Luc (Clontech, Palo Alto, California, USA) and Lipofectamine 2000 reagent (Invitrogen, Carlsbad, California, USA) at a ratio of 3:1. COS7 cells were transfected with 0.1 μg of expression plasmid and 0.4 μg of GRE-Luc using the Fugene 6 Transfection Reagent (Roche Molecular Diagnostics, Mannheim, Germany) at a ratio of 3:1. In all conditions, pCMV β-galactosidase was cotransfected, luciferase and β-galactosidase activity was determined with the Dual-Light kit (Tropix, Bedford, Massachusetts, USA), and luciferase activity was normalized to β-galactosidase activity.
eNOS activity assay and cGMP measurement. eNOS activity was determined by measuring the conversion of [3H]L-arginine to [3H]L-citrulline in the presence or absence of the competitive NOS inhibitor L-NAME (1 mM) using an NOS assay kit (Calbiochem-Novabiochem). Mouse aortae were homogenized in ice-cold PBS containing 1 mM EDTA, the homogenates were pelleted, and 5 μg of protein extracts from the supernatant were used for the eNOS assay as described (17). Unlabeled L-arginine was added to [3H]L-arginine (specific activity, 60 Ci/mmol) at a ratio of 3:1. For measurement of intracellular cGMP (26), cells were pretreated with 0.5 mM isobutylmethylxanthine for 15 minutes, and proteins were extracted with 0.1 N HCl and analyzed with a cGMP radioimmunoassay kit (Amersham International, Little Chalfont, United Kingdom).
Real-time PCR. Total RNA isolation and reverse transcription from ischemic regions were performed as described (27). Real-time PCR was conducted using Biorad iCycler (iCycler iQ Multi-Color Real Time PCR Detection System; Biorad, Hercules, California, USA). The following primer pairs were used: mouse eNOS (5′-TTCCGGCTGCCACCTGATCCTAA-3′ forward and 5′-AACATGTGTCCTTGCTCGAGGCA-3′ reverse) and mouse GAPDH (5′-GAGAATGGGAAGCTTGTCATC-3′ forward and 5′-GTCCACCACCCTGTTGCTGTA-3′ reverse). The fluorescence of Cyber green (PE Biosystems, Foster City, California, USA) was used for quantitation of amplicons. The PCR conditions for eNOS were 95°C (13:30 minutes), 94°C (5:00 minutes), and then 34 cycles of 94°C (40 seconds), 60°C (1:20 minutes), and 72°C (1:20 minutes), followed by 88°C (15 seconds) for data collection. The PCR conditions for GAPDH were 95°C (13:30 minutes), 94°C (5:00 minutes), and then 23 cycles of 94°C (1 minute), 62°C (2:00 minutes), and 72°C (2:30 minutes), followed by 83°C (15 seconds) for data collection. The relative amount of eNOS cDNA was calculated by reference to the GAPDH cDNA in the sample using the Ct method (ABI PRISM 7700 Sequence Detection System; Applied Biosystems, Foster City, California, USA) and then compared with the amount at the 0 time point. Ct is the point at which the exponential increase in signal (fluorescence) crosses a somewhat arbitrary signal level (usually 10 times background) and is related to the copy number by 2–Ct.
Vascular ring bioassay. Wall tension was measured as described (28). Briefly, isolated aortae were cut into rings (1.5–2 mm) and mounted on an isometric myograph (610M, Danish Myo Technology, Aarhus, Denmark) in physiological solution (118 mM NaCl, 4.6 mM KCl, 25 mM NaHCO3, 1.2 mM MgSO4, 1.2 mM KH2PO4, 1.25 mM CaCl2, 10 mM glucose, 0.025 mM EDTA) at 37°C with or without dexamethasone for 60 minutes. After equilibration, contractile responses were recorded using a data-acquisition and —recording software (Myodaq and Myodata, Danish Myo Technology).
Transient middle-cerebral-artery occlusion. All experiments were conducted in accordance with National Institute of Health and Massachusetts General Hospital institutional guidelines. The left middle cerebral artery of male SV-129 (Taconic, Germantown, New York, USA) or eNOS–/– mice was occluded for 2 hours by advancing an 8-0 nylon silicone-coated monofilament 10 mm distal to the carotid bifurcation as described (14). The infarcted areas were measured on brain sections stained with 2,3,5-triphenyltetrazolium (TTC; 24 hours) or on hematoxylin and eosin–stained cryostat sections (72 hours) with an image-analysis system (Bioquant IV, R&M Biometrics, Nashville, Tennessee, USA) and were quantitated by summing the volumes of each section. In some experiments, infarct volume was corrected for edema (29). Regional CBF was determined by laser Doppler flowmetry (Perimed PF2B, Stockholm, Sweden) and recorded on a MacLab/8 data acquisition system (AD Instruments, Milford, Massachusetts, USA) as described previously. Serum dexamethasone levels were determined by Quest Diagnostics Inc (San Juan Capistrano, California, USA).
CBF measurement. Regional CBF was quantified using the [14C]iodoantipyrine autoradiography technique as described previously (30). Isoflurane-anesthetized mice received [14C]iodoantipyrine (5 μCi in 100 μl saline) for 1 minute by infusion. Arterial blood samples were collected at 5-second intervals onto preweighed filter paper disks and measured by liquid scintillation spectrometry (RackBeta 1209; Pharmacia-Wallac, Gaithersburg, Maryland, USA). Immediately after decapitation, the whole head was immersed in chilled isopentane over dry ice (–45°C), and then the frozen brains were sectioned coronally (20 μm). The sections were thaw-mounted on glass coverslips, dried (60°C), and exposed to autoradiography along with [14C]polymer standards (Amersham Life Sciences Inc.). In some experiments, CBF was determined using the indicator fractionation technique with N-isopropyl-[methyl 1,3-14C]-p-iodoamphetamine as described previously (17).
Statistics. The difference in cerebral infarct volume was analyzed by one-way ANOVA followed by Duncan's test. CBF and ring experiments were analyzed by two-way ANOVA followed by t-test or Tukey test, respectively. Student's t test was used for all other analyses. All values are expressed as means ± SEM unless otherwise indicated.
Acute neuroprotective effects of corticosteroids mediated by eNOS. To determine whether acute administration of corticosteroids protects against ischemic stroke, mice were treated with the synthetic corticosteroid dexamethasone and subjected to transient middle cerebral-artery occlusion (MCAo). In a dose-dependent manner, dexamethasone treatment 1 hour before and 2 hours after MCAo decreased cerebral infarct volume by 32% as compared with vehicle treatment (74 ± 8 mm3 vs. 108 ± 9 mm3, P < 0.05). The reduction in stroke volume by dexamethasone was evident up to 3 days after ischemia, suggesting sustained neuroprotection by corticosteroids (Figure 1a).
Acute neuroprotection by corticosteroids requires eNOS. (a) Dose–dependent effects of dexamethasone (Dex) on stroke volume derived from TTC-stained (24 hours) or hematoxylin and eosin–stained (72 hours) brain sections after MCAo (n = 8–11). *P < 0.05 vs. vehicle. (b) Effects of delayed steroid treatment on neuroprotection. Shown is a schema of treatment regimens and corresponding cerebral infarct reduction by dexamethasone relative to the vehicle (Veh) group (n = 7–11). *P < 0.05 vs. vehicle. (c) Infarct volume corrected for edema in wild-type and eNOS–/– mice treated with vehicle or dexamethasone (20 mg/kg bolus administered intraperitoneally [i.p.]) 24 hours after MCAo (n = 5–11). Wild-type mice are identical to those in (a). *P < 0.05 vs. all other groups; WT, wild-type. (d) The eNOS protein and mRNA levels after steroid treatment. Equal amounts of aortic tissue homogenates (40 μg) were immunoblotted against eNOS followed by stripping and reprobing against actin (top panels). A representative experiment is shown. Three separate experiments yielded similar results. Brain eNOS mRNA expression from mice treated with vehicle or dexamethasone with or without RU486 (RU) or LY294002 (LY) for 24 hours is shown in the lower panel. The corrected eNOS mRNA levels (eNOS/GAPDH) were determined by real time PCR (n ≥ 3 for each condition). t, time.
To evaluate neuroprotection by corticosteroids after the onset of ischemia, we compared the efficacy of different treatment regimens to reduce stroke size (schema in Figure 1b). The standard regimen was compared to a single bolus injection of dexamethasone 2 or 4 hours after the onset of ischemia. Administration of dexamethasone up to 2 hours after the onset of ischemia reduced stroke volume by 25% as compared with the vehicle group (84 ± 7 mm3 vs. 112 ± 8 mm3, P < 0.05), an effect that was lost when treatment was initiated 4 hours after MCAo (Figure 1b). Thus, corticosteroids rapidly confer neuroprotection when administered early after the onset of ischemia.
Reduction in myocardial infarct size by steroids involves the nontranscriptional activation of eNOS (23). To test whether eNOS mediates the acute cerebrovascular protective effect of dexamethasone, cerebral infarct volume was evaluated in mice with a targeted disruption of the eNOS gene (eNOS–/–) (31). In contrast to the wild-type mice treated with high-dose steroids, eNOS–/– mice treated with dexamethasone showed no reduction in stroke size (reduction, 4% ± 10%; P > 0.05) after transient ischemia (Figure 1c), whereas physiological parameters were comparable (data not shown). These findings indicate that eNOS mediates the acute neuroprotective effects of corticosteroids. Although eNOS is required for neuroprotection by steroids, eNOS mRNA and protein levels in brain tissues were unchanged in vivo, suggesting a nontranscriptional mechanism of eNOS enhancement (Figure 1d).
Corticosteroids have been thought to ameliorate ischemic tissue damage by reducing associated brain edema (18). Nevertheless, when stroke volume was corrected for brain edema (29), the protective effect of dexamethasone was sustained (65 ± 8 mm3 vs. 92 ± 7 mm3, n = 11, P < 0.05), indicating that corticosteroids protected against cerebral injury (Figure 1c). It is interesting to note that we also found an eNOS-dependent effect of steroids on edema formation. Dexamethasone reduced cerebral edema by 37% in the eNOS–/– mice as compared with 72% in the wild-type strain (17± 3 mm3 [n = 10] vs. 5 ±1 mm3 [n = 11], P < 0.05).
Corticosteroids enhance NO-dependent CBF. Because cerebral infarct size is inversely related to CBF, and endothelium-derived NO regulates cerebral vascular tone and CBF (32), we examined whether corticosteroids can enhance regional CBF to ischemic areas of the brain. Mice were subjected to transient MCAo and regional intraischemic CBF was determined using [14C]iodoantipyrine autoradiography. After MCAo, there was little or no blood flow in the cortical and subcortical parietal lobe of vehicle-treated mice. In mice treated with dexamethasone, the entire core infarct zone appeared smaller, and blood flow was substantially higher in the core region, particularly in the caudate putamen and in the adjacent peri-ischemic areas (Figure 2a). These findings demonstrate that corticosteroids can enhance intraischemic CBF.
Regulation of CBF by corticosteroids.(a) Representative pseudocolor image of CBF ([14C]iodoantipyrine autoradiography) from coronal brain sections 1 hour after MCAo in mice treated with dexamethasone (20 mg/kg) or vehicle. (b) Regional changes in absolute CBF ([14C]iodoantipyrine) in ipsilateral and contralateral brain areas of mice treated with dexamethasone (20 mg/kg administered intraperitoneally) or vehicle (n = 10 per group). (c) Changes in absolute CBF in eNOS–/– mice were determined by the [14C]iodoamphetamine technique and expressed as percentages of contralateral CBF of vehicle-treated mice. *P < 0.05 vs. vehicle.
To confirm that corticosteroids augment regional CBF, absolute CBF was quantified by [14C]iodoantipyrine measurement. Within 1 hour, treatment with dexamethasone increased absolute CBF in the core ischemic zone (caudate putamen and thalamus) and the penumbral areas (Figure 2b). It is interesting to note that corticosteroids also enhanced CBF in nonischemic areas of the brain. Since equal percentages of ischemia between treatment groups were confirmed by laser Doppler flowmetry (Table 1 and Methods), this demonstrates a beneficial effect of corticosteroids on absolute ischemic CBF. In addition, it provides evidence that enhancement of cerebral perfusion by steroids occurs rapidly, suggesting a nongenomic mechanism of action. Indeed, the increase in CBF by corticosteroids was mediated by NO, since no augmentation of CBF was observed in either the ischemic or nonischemic hemispheres after MCAo in eNOS–/– mice (Figure 2c).
Effects of high-dose corticosteroids on SV-129 mice
Nontranscriptional activation of PI3K mediates stroke protection by corticosteroids. Endothelial NO release is enhanced through direct phosphorylation of eNOS by the protein kinase Akt downstream of PI3K (33, 34). We have previously shown that GR activates eNOS through the PI3K/Akt pathway in a nontranscriptional fashion (23). To determine whether rapid GR signaling to PI3K mediates neuroprotection by steroids, we evaluated cerebral infarct volume with the PI3K inhibitor, LY294002, and the GR antagonist RU486. As compared with the high-dose treatment group, LY294002 and RU486 both reversed the beneficial effects of dexamethasone on cerebral infarct volume without altering baseline values, suggesting that neuroprotection is mediated by PI3K and GR (Figure 3a). Comparison of physiological parameters, mortality, and serum dexamethasone levels after treatment revealed no significant difference between treatment groups (Table 1).
Activation of PI3K/Akt mediates neuroprotection. (a) Neuroprotection by dexamethasone is mediated by GR and PI3K. Cerebral infarct volume after MCAo in response to dexamethasone (20 mg/kg), RU486 (200 mg/kg) and LY294002 (5 mg/kg, n = 7–11). Vehicle and dexamethasone groups are identical to those in figure 1a. *P < 0.05 vs. vehicle, **P < 0.05 vs. dexamethasone. RU, RU486; LY, LY294002. (b) In vivo activation of PI3K by dexamethasone. PI3K activity from mouse brain tissue immunoprecipitated with GR or p85 antibody. The product of the kinase reaction, PIP3, was visualized by autoradiography. (c) The left panel shows Akt activity and total Akt from mouse brain tissue. Mice were treated with vehicle or dexamethasone, with or without actinomycin D (ActD) (5 mg/kg), RU486, or LY294002. The right panel shows P-Akt (S473) and total Akt from mouse aortae. P, phosphorylation form of the protein. Representative experiments are shown.
Class I PI3K catalyzes the formation of 3′-phosphorylated phosphatidylinositols (e.g., phosphatidylinositol 3,4,5-trisphosphate [PIP3]) from phosphatidylinositol phosphate precursors (35). This leads to the subsequent recruitment and activation of protein kinase Akt (36). To evaluate the rapid signaling mechanisms initiated by GR in vivo, we measured GR-associated PI3K activity from brains of dexamethasone-treated mice. Dexamethasone treatment leads to a rapid increase in PI3K activity, as measured by the generation of PIP3. GR-associated PI3K activity was blocked by cotreatment with the PI3K inhibitor LY294002, and the GR antagonist RU486, suggesting that the effect of dexamethasone was mediated by GR (Figure 3b).
Furthermore, steroid treatment leads to in vivo activation of Akt downstream of GR and PI3K, as determined by Akt kinase activity (upper panels in Figure 3c) and Akt phosphorylation (lower panels in Figure 3c). Of note, the transcriptional inhibitor actinomycin D, in doses reported to inhibit transcriptional responses to thyroid hormone and dexamethasone in vivo (8, 37), did not inhibit activation of Akt by corticosteroids. Thus, activation of the PI3K/Akt axis in vivo does not require induction of gene expression by GR. These findings suggest that activation of the PI3K/Akt axis mediates acute neuroprotection by steroids.
Regulation of vascular tone by corticosteroids. Blood flow is partially determined by peripheral vascular resistance, which is negatively regulated by endothelium-derived NO. To determine the effects of acute administration of corticosteroids on vascular tone, we measured wall tension of vascular rings. In vitro, the α-adrenergic agonist phenylephrine caused dose-dependent contraction of aortic rings, which can be reversed by acetylcholine (ACh) in an NO-dependent manner. Cotreatment of aortic rings with relatively high doses of dexamethasone reduced aortic-ring wall tension (Figure 4, a and b), which was reversed by RU486 and LY294002 (Figure 4c). Furthermore, in the absence of eNOS activity, due to either inhibition by L-NAME or lack of enzyme in eNOS–/– tissue, baseline contraction was markedly increased and remained unchanged by dexamethasone or ACh. These data demonstrate that dexamethasone exerts rapid, NO-dependent effects and suggest that vasorelaxation by dexamethasone requires activation of eNOS through GR and PI3K.
NO-dependent vasodilation by corticosteroids. (a) Wall tension to cumulative concentrations of phenylephrine in aortic rings incubated with 1 μM dexamethasone (n ≥ 6). (b) Dose-dependent effects of dexamethasone on wall tension in aortic rings pretreated with 1 μM phenylephrine (n ≥ 4). (c) Wall tension in aortic rings from wild-type or eNOS–/– mice treated with phenylephrine and dexamethasone or ACh (3μM) with or without RU486, LY294002, or L-NAME (n ≥ 4). (d) Aortic eNOS activity from wild-type or eNOS–/– mice treated with vehicle or dexamethasone with or without RU486 or LY294002 for 2 hours. Three separate experiments yielded similar results. *P < 0.05.
To determine whether corticosteroids actually stimulate eNOS in vivo, mice were treated with dexamethasone, and aortic NOS activity was measured by the enzymatic conversion of the eNOS substrate L -arginine to L -citrulline (Figure 4d). Dexamethasone treatment induced a threefold increase in eNOS activity, which was completely blocked by cotreatment with LY294002 and RU486. In contrast, baseline NOS activity was virtually absent in eNOS–/– mice and remained unresponsive to dexamethasone stimulation (data not shown), indicating that GR specifically enhances eNOS activity through a PI3K-dependent mechanism in vivo.
Nontranscriptional actions of GR require high-dose corticosteroids. Dexamethasone activates eNOS in human endothelial cells (ECs) in a nontranscriptional fashion, which is mediated by the PI3K/Akt pathway (23). Generation of NO increases intracellular cGMP through stimulation of guanylate cyclase. To confirm that dexamethasone stimulates NO production in ECs from different vascular domains, we measured cGMP levels (Figure 5a). Dexamethasone increased cGMP levels about twofold in human umbilical-vein ECs (HUVECs), an effect which was reversed by LY294002 and the NOS inhibitor L-NAME. Similar effects were observed in human aortic ECs (HAECs). We also compared the ability of different corticosteroids to activate eNOS (Figure 5b). All of the tested corticosteroids, including the endogenous steroid hydrocortisone, were found to activate eNOS, demonstrating a corticosteroid class effect on eNOS activity.
Nontranscriptional activation of eNOS by high-dose corticosteroids. (a) Shown are cGMP levels in human ECs after dexamethasone treatment. The upper panel shows cGMP levels in HUVECs stimulated with S-nitrosoglutathione (GSNO, 300 μM) or dexamethasone (100 nM) with or without LY294002 (LY, 30 μM) or L-NAME (1 mM, n = 4). The lower panel shows cGMP levels in HAECs stimulated with dexamethasone with or without LY294002 (n = 4). Ctrl, control. *P < 0.05 vs. control. GSNO, S-nitrosoglutathione; LY, LY294002. (b) Effects of 100 nM of dexamethasone, prednisolone, hydrocortisone, or triamcinolone acetonide on eNOS activity in HUVECs (n = 2). Pred, prednisolone; HC, hydrocortisone; TA, triamcinolone acetonide. (c) Dose–response effects of dexamethasone on eNOS activity (HUVECs, n = 5) and GRE-luciferase activity (bovine aortic endothelial cells transfected with pGRE-luc, n = 6). *P < 0.01 vs. control. (d) Dexamethasone serum levels in mice after intraperitoneal bolus administration of low- and high-dose dexamethasone.
Nontranscriptional activation of PI3K in ECs requires high doses of dexamethasone (23). Furthermore, only high-dose steroid treatment provides neuroprotection. This is in contrast to the genomic responses to steroids. To further characterize the transcriptional and nontranscriptional actions of GR, we compared the dose–response data for eNOS activity with results from a glucocorticoid-responsive element–dependent (GRE-dependent) luciferase reporter. Although maximal induction of eNOS activity required doses of 100 nM to 1 μM of dexamethasone, much lower doses of dexamethasone (10 nM) readily induced maximal GRE-mediated transcriptional activity. This was reflected in an approximately 10-fold difference in EC50 values (1.1 nM for GRE-reporter vs. 13 nM for eNOS activity) and ECmax values (10 nM for GRE-reporter vs. 100 nM for eNOS activity), as derived from logarithmically transformed curve-fit analysis (Figure 5c).
Consistent with our in vitro findings, the peak serum dexamethasone level in mice after administration of neuroprotective doses (20 mg/kg bolus administered intraperitoneally) was approximately 140 nM (5.5 μg/dl) (Figure 5d). No neuroprotection was observed when mice were given 0.2 mg/kg of intraperitoneal dexamethasone, which gave a peak dexamethasone level of 33 nM (1.3 μg/dl). Together, these findings indicate that pharmacologic concentrations of corticosteroids are required for nontranscriptional activation of eNOS and NO-mediated neuroprotection. Despite such high concentrations of corticosteroids, the observed effects are relatively specific since the neuroprotective effects of corticosteroids are absent in eNOS–/– mice and could be blocked in vivo by inhibitors of GR and PI3K.
GR mediates the non-nuclear activation of PI3K/Akt by corticosteroids. To determine on a molecular level whether GR mediates PI3K activation, we performed studies in COS7 cells that lack endogenous GR (38). In mock-transfected cells, treatment with dexamethasone did not activate PI3K or Akt, whereas transfection of human GR enabled activation of the PI3K/Akt pathway by dexamethasone. Furthermore, transfection of a dimerization-defective GR mutant (A458T), which is unable to bind DNA and transactivate target genes (38), still activated the PI3K/Akt pathway in a ligand-dependent manner (upper panels in Figure 6a). These results suggest a nontranscriptional mechanism of PI3K/Akt activation, since GR mutant A458T was unable to induce significant transcriptional activity (lower panel in Figure 6a).
Nontranscriptional activation of PI3K by GR. (a) Nontranscriptional activation of PI3K and Akt kinase by GR: The upper panels show PI3K and Akt kinase assays in COS7 cells transfected with expression plasmids encoding GRα (GRSB) or dimerization-defective GR (A458T). Lysates were immunoblotted against GR. The lower panel shows GRE-reporter activity of COS7 cells cotransfected with either GRSB or A458T expression plasmid with GRE-luc and stimulated with dexamethasone (100 nM). (b) MEFs obtained from wild-type, GR–/–, and dimerization-defective GRdim mice were assayed for Akt activity and expression of GR by Western blotting. (c) Dexamethasone activates the PI3K/Akt pathway through GR. The upper panel shows immunoprecipitation of ERα, GR, or MR followed by PI3K assay or immunoblotting against ERα, GR, and MR. (d) Akt kinase assay in COS7 cells transfected with expression plasmids encoding ER, GR or MR. Lysates of mock-transfected or receptor-transfected cells were immunoblotted against ER, GR, or MR. Representative experiments are shown; m, mock transfected. WB, Western blot; P-GSK-3, phospho-GSK-3.
Nontranscriptional activation of PI3K was further evaluated in mouse embryonic fibroblasts (MEFs) from genetically modified mice (Figure 6b). While dexamethasone did not activate Akt in GR–/– MEFs, activation was readily observed in wild-type MEFs, expressing endogenous levels of GR, and in cells with a dimerization-defective GR (GRdim) that renders GR unable to bind to DNA and induce gene expression (5). These results indicate that GR is required for nontranscriptional activation of PI3K and downstream effectors. These findings also suggest specific recruitment of PI3K to GR. Indeed, we have recently described the association of GR with the p85 α regulatory subunit of PI3K in a ligand-dependent manner (23). Corticosteroids can bind to and activate the mineralocorticoid receptor (MR). To determine whether the nontranscriptional effects of dexamethasone were specific for GR activation, PI3K activity associated with different steroid receptors was examined in HUVECs (Figure 6c). Treatment with dexamethasone readily stimulated GR-associated PI3K activity but did not stimulate association of PI3K activity with either MR or ERα, suggesting that PI3K specifically associates with ligand-bound GR.
To corroborate the specificity of high-dose dexamethasone action toward GR, we evaluated Akt activation in COS7 cells transfected with expression plasmids for ERα, GRα, or MR (Figure 6d). Although all receptors were sufficiently expressed after transfection, only GR-mediated activation of Akt by dexamethasone. These findings suggest that although high doses of steroids are required for non-nuclear actions, the effects are nevertheless specifically mediated by GR.
Clinical studies of corticosteroids in stroke have yielded conflicting results, which may reflect differences in dosage regimens, the time to onset of treatment, or the specific clinical condition. Although several trials in patients with presumed ischemic stroke have failed to show a reduction in mortality (39–42), at least one study has demonstrated a significant improvement in functional outcome (43). In experimental stroke, physiological levels of corticosteroids increase the severity of ischemic injury after global ischemia (44). However, our finding that corticosteroids are neuroprotective is consistent with previous studies showing reduction in stroke size after focal cerebral ischemia (20–22). Similar to these earlier studies, we found that pharmacological concentrations of corticosteroids were required for acute neuroprotection, whereas lower doses had no effect. It is interesting to note that this dose–response relationship has also been observed in spinal cord injury, where low-dose corticosteroids are not protective, whereas high doses improve clinical outcome when given within the first 8 hours after injury (45, 46).
Several lines of evidence support the conclusion that the neuroprotective effect of high-dose steroids is mediated by nontranscriptional activation of eNOS through the PI3K/Akt pathway. Rapid augmentation of ischemic CBF and stroke protection by steroids is mediated by eNOS, since these effects are reversed in the absence of eNOS. Although neuroprotection is mediated by eNOS, eNOS expression levels were unchanged by steroid treatment in our studies (i.e., <24 hours), suggesting nontranscriptional enhancement of eNOS activity. This is in contrast to a previous report showing that longer-term steroid therapy (i.e., >72 hours) leads to the downregulation of eNOS and an increase in systemic blood pressure (24). Thus, the beneficial and detrimental effects of steroid therapy with regard to eNOS expression are, in part, dependent on the duration and dose of treatment. In addition, two previous studies suggest that injury from MCAo could lead to increased eNOS expression (47, 48). This is in contrast to the findings of our study, which did not show significant change in eNOS expression in the brain after MCAo in any treatment group. Differences between our study and previous studies include the animal species (i.e., mouse vs. rat), the model of cerebral ischemia (transient vs. permanent occlusion), and the method of measuring eNOS expression (real time PCR vs. immunostaining or semiquantitative RT-PCR).
We found that inhibition of PI3K reversed the beneficial effects of dexamethasone on stroke. Steroids activate eNOS in vivo and in vitro through the PI3K/Akt pathway. Although the activation of PI3K by corticosteroids requires GR, no GRE-mediated gene transcription is necessary for PI3K activation. Instead, activation of PI3K involves the association of the regulatory p85 subunit of PI3K with GR (23). This non-nuclear effect of GR is similar to that of the estrogen receptor, which interacts with and activates PI3K in a ligand-dependent manner (11). Furthermore, the peak serum dexamethasone level in mice treated with neuroprotective doses corresponded to the in vitro concentration that maximally stimulated eNOS and PI3K. Together, these data strongly suggest that nontranscriptional actions of GR mediate neuroprotection through activation of PI3K and eNOS, leading to increases in CBF.
In our studies, the non-nuclear and protective actions of GR require steroid doses at least 10 times as high as those required for the genomic response by the receptor. Yet, these nongenomic actions are specifically mediated by GR. Clearly, further studies are needed to address this discrepancy between transcriptional and nontranscriptional actions. One might speculate that activation of PI3K requires so-far-undiscovered adaptor molecules bound to a subset of GR that alter the binding affinity of the receptor for steroids.
Postischemic inflammation and microvascular occlusion contribute to ischemic brain injury, which can be counteracted by endothelial-derived NO (13, 49). The inhibitory effects of corticosteroids on endothelial-leukocyte adhesion are mediated by GR (50). However, the mechanism by which GR inhibits endothelial-leukocyte interaction is not well understood. By enhancing endothelial NO production, corticosteroids might inhibit platelet aggregation and leukocyte adhesion, thereby enhancing blood flow. The features of neuroprotection by GR resemble the role of NO in stroke protection, since endothelial-derived NO decreases stroke size by improving regional CBF (14). On the other hand, NO donors lose their protective efficacy when administered more than 2 hours after ischemia, an effect that is similar to the effects of dexamethasone (32, 51). Indeed, the ability of dexamethasone to induce NO-dependent relaxation in the mouse aorta probably also contributes to its ability to increase eNOS activity and CBF in the nonischemic regions of the brain. This is a novel feature of corticosteroids that is distinct from its anti-inflammatory effects.
Protection from ischemia in our model is primarily dependent on CBF. However, in a different setting, non-nuclear activation of PI3K and Akt by GR could exert perfusion-independent protective effects, since Akt directly inhibits apoptosis in neural tissue and the myocardium (52, 53). We have recently shown that high-dose corticosteroids decrease myocardial infarct size through the PI3K/Akt pathway (23). Although steroid therapy reduced myocardial infarct size, there was no change in the area at risk between treatment groups, suggesting additional blood flow–independent effects of corticosteroids on ischemic myocardium. Indeed, corticosteroids can promote survival through an Akt-related protein kinase, serum- and glucocorticoid-regulated kinase (SGK) (54, 55). Because SGK is a downstream target of PI3K (56), the ability of ligand-bound GR to activate PI3K may, therefore, provide an additional mechanism by which corticosteroids can regulate survival.
The therapeutic use of corticosteroids is limited by side effects, which occur at relatively low concentrations and are typically associated with long-term treatment. Thus, it appears that these detrimental effects might be related to the genomic actions of corticosteroids. By linking GR to PI3K, the biological and pharmacological actions of corticosteroids could be considerably broadened. This would require the development of a novel class of drugs that selectively activate the nontranscriptional actions of GR. Indeed, synthetic compounds that separate specific GR functions, such as transactivation from transrepression, have recently been identified (57).
We thank R. Mandle and F. Rehman (Center for Blood Research, Boston, Massachusetts, USA) for assistance in performing real-time PCR. Plasmids GRSB and A458T were kindly provided by A.C.B. Cato (Forschungszentrum Karlsruhe, Karlsruhe, Germany), expression plasmids for human GR and MR by R. Evans (Salk Institute, La Jolla, California, USA), and human ERα by W.W. Chin (Eli Lilly and Co., Indianapolis, Indiana, USA). This work was supported by grants from the National Institutes of Health (HL-70274, HL-52233, HL-48743, and NS-10828), the American Heart Association Bugher Foundation Award, the Ministero per l'Università e la Ricerca Scientifica e Tecnologica (MURST), the Deutsche Forschungsgemeinschaft (Li-948 1/1 to F.P. Limbourg), and the American Heart Association, New England Affiliate Inc. (to J.-C. Plumier).
Florian P. Limbourg and Zhihong Huang contributed equally to this work.
Conflict of interest: The authors have declared that no conflict of interest exists.
Nonstandard abbreviations used: glucocorticoid receptor (GR); endothelial nitric oxide synthase (eNOS); neuronal nitric oxide synthase (nNOS); cerebral blood flow (CBF); phosphatidylinositol 3-kinase (PI3K); mitogen-activated protein kinase (MAPK); estrogen receptor α (ERα); Nω-nitro-l-arginine methyl ester (l-NAME); bovine aortic endothelial cells (BAECs); 2,3,5-triphenyltetrazolium (TTC); middle-cerebral-artery occlusion (MCAo); phosphatidylinositol 3,4,5-trisphosphate (PIP3); acetylcholine (ACh); endothelial cells (ECs); human umbilical-vein ECs (HUVECs); human aortic ECs (HAECs); glucocorticoid-responsive element (GRE); mouse embryonic fibroblasts (MEFs); mineralocorticoid receptor (MR); serum- and glucocorticoid-regulated kinase (SGK).
Copyright © 2014
American Society for Clinical Investigation


text:
No class on Tues. Sept. 23rd.
Makeup class on Friday, Sept. 26 at 2:30pm in Marcus 106.
WinDLX InformationComputer Architecture Educational Tools
Technical papers (Copyright Note):
Additional Resources for the recommended textbook (select "Resources"and then "Companion Material").
The Computer Architecture Home Page.
Additional Links
Created by Prof. Israel Koren, koren 'at' ecs.umass.edu

subset: commoncrawl


text:
Edited Kentucky Downs release
Thoroughbred racing returns to Kentucky Downs in Franklin, Ky., on Saturday, Sept. 16 for a six day racing season, which concludes on Tuesday, Sept. 26. The highlight of the meet is the $450,000 Kentucky Cup Turf Festival, featuring four stakes races on Saturday, Sept. 23. The grade III, $200,000 Kentucky Cup Turf, to be run on Turf Festival Day, is the showcase race of the season. Last year, Kentucky-based Silverfoot pulled away down the stretch for a decisive win in the Kentucky Cup Turf. The 2001 Turf was won by Chorwon and the $186,000 top prize boosted his career earnings over the $1 million mark. Chorwon and Silverfoot both competed in the $2-million John Deere Breeders' Cup Turf (gr. IT) in their next starts. The six-race Turf Festival card also includes the $50,000 Yaqthan Stakes, the $100,000 Kentucky Cup Turf Dash and the $100,000 Kentucky Cup Ladies Turf. Sand Springs, the winner of the 2004 Ladies, later won the grade I, $500,000 Diana Stakes at Saratoga Race Course. Battle Won, the winner of the 2004 Sprint at Kentucky Downs, also moved to the top of the national standings following his win at Kentucky Downs. Battle Won won the grade II Churchill Downs Handicap. Also in 2004, jockey Rafael Bejarano left his mark on Kentucky Downs while on his way to being the nation's leading jockey in wins. Bejarano won the leading jockey honors and also won six races, a Kentucky Downs record, on one card. Last year, Bejarano was based in New York, but came to Kentucky Downs on Turf Festival day and recorded three stakes wins. Kentucky Downs will run Sept. 16, 18 and 19 and resume for closing weekend, Sept. 23, 25 and 26. The two Saturday cards will be run in conjunction with the races from Turfway Park. Saturday post-times will be 12:10 p.m. (CDT).On opening day, Sept. 16, Kentucky Downs will feature the $40,000 Pleasant Temper Stakes. Fans will also be treated to the 3rd Annual Kentucky Downs Wiener Dog Challenge. The charity event will crown the region's fastest dachshund and all proceeds will go to the Simpson County Animal Shelter. Also on Saturday, Sept. 16, the first 1,000 patrons will receive an embroidered Kentucky Downs souvenir baseball hat. All giveaways are with clubhouse admission.Kentucky Downs will run nine live races on Monday, Sept. 18 and Monday, Sept. 25. Eight live races will be held on Tuesday, Sept. 19 and Tuesday, Sept. 26. Post-time on Monday and Tuesday is 1:45 p.m. CDT. Located on the Kentucky/Tennessee border, Kentucky Downs was originally opened as Dueling Grounds Racecourse. The only "European style," all turf racing Thoroughbred facility in the United States, Kentucky Downs is now owned by Kelley Farms, Turfway Park and Churchill Downs.
Kentucky Cup Turf Festival Highlights Kentucky Downs Meet
Most Popular Stories
- Game On Dude, Won Three Big 'Caps, Retired
- Sherman Says 'Chrome' Looks as Good as Ever
- Top Filly Princess of Sylmar Retired
- California Chrome Draws Rail in PA Derby
- Big Gains Mark Keeneland Sale's Day 11
- Undrafted Returns in Kentucky Downs Turf Dash
- Illinois Board Has Tough Call on 2015 Dates
- Filly First Winner for Line of David
- Pedigree Analysis: Inbreeding Trends
- Top Stallion Street Cry Dead at Age 16


text:
girls who were bullied most of their life and gain confidence at one point should be feared most because they dont take anyone's shit no longer and they will destroy you if you think otherwise
my life got about a thousand times better once i stopped censoring myself
and by censoring i don't mean i suddenly embraced indiscriminate swearing; i mean i stopped trying to sugarcoat my past or my feelings; i stopped lying by omission; i stopped having guilty pleasures; i began unabashedly enjoying whatever i liked; i became very honest; i cut out of my life poisonous people and negative ideals, and i am so, so much happier for it
Do you ever think about the fact that the US has created and legitimized a system of institutionalized inequality by funding schools through property taxes? That basically a child's education is only as good as the value of the property in their neighborhood. Funny how education is so often viewed as an equalizing factor when there is nothing equal about it.
Not all Americans:
• are fat
• like McDonald's
• like meat
• drink coffee
All Americans:
• PouR THEIR TEA IN THE BOSTON HARBOR
(Source: hipsteryoghurt)


text:
WHO'S GOING TO RIOT FEST 2014? Hi my names Khadija. I'm fueled by cheese and grapes as well as sarcasm and I like stimulating conversations and movies and being outside and marijuana and run on sentences.
Instagram: deejlah
Instagram: deejlah
Find better matches with our advanced
matching system
19 • F • Lake Bluff, IL
Restore your account to continue meeting new people.
We'll email you a link to reset it.
An email is on its way to you. If you don't see it, try checking your Spam folder.


text:
During Oklahoma's 41-20 victory over Texas Tech, Sooners defensive back Aaron Colvin tipped a pass. Javon Harris intercepted it returned it for a touchdown. Soon after, the Texas Tech crowd began to make its way out of Jones Stadium with 8:40 remaining in the third quarter. The interception was Harris' second of this season and the fifth of his career. The pick-six was the first of his career. Harris sat down with The Oklahoman after the victory.
Oklahoma football: Q&A with defensive back Javon Harris
Was that a good feeling to score a touchdown and see all the fans walk out of the stadium?
Ha. Yeah, it was a good feeling but I have to give credit to (Colvin) on that play. He had the pass deflection. I just took advantage of the opportunity of the ball being in the air to take it in.
What did it feel like to go on the road and put away a team in the third quarter?
We just wanted to come out, start fast, finish fast from halftime until the end of the game. You gotta finish a game out strong and to put the game away the way we did is great. We just have to continue to work on everything we've been working on and we played good.
On that touchdown, what went through your head specifically when you saw it coming?
Make sure I catch the ball. The one thing we've been working on is these turnovers and making sure I have the ball and then just take it as far as I could. All I saw was the end zone when I caught it.
Trending Now
- 53702Oklahoma beheading: Prosecutors to seek death penalty for attack suspect
- 18697OSU basketball: Five-star guard Jawun Evans commits to Cowboys
- 17175Oklahoma Beheading: Neighbors say murder defendant in Oklahoma beheading case was 'odd' and 'quiet'
- 16220Oklahoma beheading: Murder defendant confessed, attempted second beheading
- 13685Oklahoma beheading: Friend remembers Colleen Hufford
- 13561Oklahoma football: Alex Ross has unique personality, talent
- 12281OKC Thunder: Mitch McGary brings infectious enthusiasm


text:
Hail Mary in a Song- Jorge L Reyes- Catholic Christian Music
Version 8 and above required to view videos
Uploaded by on Dec 21, 2012
The Hail Mary in a song, I hope you like it, sing it along with me to Mary, Jesus loves you!
Mary Catholic.music
0 Comments
Leave a Comment
Suggested Videos
Catholic Worship Podcast Episode 8 - ... 7,606 views View Video
Catholic Worship Podcast Episode 3 - Divine ... 6,958 views View Video
Catholic Worship Podcast Episode 2 - Tantum ... 6,987 views View Video
Hail Mary 8,166 views View Video
Mary Ann Glendon named New U.S. Ambassador ... 8,333 views View Video
Want to share your informative and entertaining video with an international audience? Sign In
Most Popular
WARNING GRAPHIC PHOTOS (RAW) - ISIS begins killing Christians in Mosul, CHILDREN BEHEADED Read More
Islamic State beheads journalist James Foley (WARNING, GRAPHIC VIDEO) Read More
WARNING GRAPHIC VIDEO - Jihadi John beheads David Haines (FULL VIDEO) Read More
WARNING GRAPHIC, RAW PHOTOS -- ISIS on Christians: 'There is nothing to give them but the sword' Read More
Steven Sotloff beheading video, uncensored (WARNING GRAPHIC VIDEO) Read More


text:
Is MLS risking several rounds of successful expansion by focusing too heavily on the New York and Los Angeles markets?
Soccer News
Soccer Scores: Anderlecht 0 – Borussia Dortmund 3
The soccer scores for USMNT players in Europe and Mexico, with Sacha Kljestan and Anderlecht in the Champions League. Read More
Soccer News: PSG and winning every game
PSG got a big win in the Champions League over Barcelona, but the concern is all the games they're not winning in Wednesday's soccer news. Read More
Soccer TV: Krasnodar vs Everton and Spurs vs Besiktas
Krasnodar host Everton and Spurs play Besiktas in the Europa League on Fox Sports 1 on Thursday's soccer TV schedule. Read More
Did MLS fail Chivas USA?
MLS commissioner Don Garber's public statements concerning Chivas USA suggests that the league could've done more for the club. Read More
Jonathan Spector out for Birmingham City with thigh injury
Birmingham City defender Jonathan Spector injured his thigh against Fulham and will miss time with the Championship club. Read More


text:
Review for October
(#) midnightblackparade 2011-10-04 11:52:57 AM
AMAZING .. AGAIN!! Keep up the good work
Author's response
Awhh, thank you! I'm writing up a new chapter now, should go up tomorrow sometime! :) xx
Log in!Register Lost password
You won't see stories with a lower score when you browse or search. Log in to adjust filter.
Featured Story
Harry Potter and the Narrative Causalityby Clell65619 (R)
Hermione discovers the power Harry has that the Dark Lord knows not. Harry enlists the aid of a cert...
In: Harry Potter
Site Stats
- Authors: 657250
- Stories: 39954
Recent Stories
The Werewolf & The Reaporby angelrose1205 (G)
In: Horror
Dragon Ball Heroes: A New Generation of Heroesby TheOtakuFox43 (R)
With his future in shambles, Trunks only has one option left after his time machine is stolen by Cel...
In: Dragon Ball Z
Humble Administrator's Backyard: a unique back gardenby bailrepair5 (PG-13)
In: Over There
Speed of Lyghtby J_Linz (PG-13)
Tabitha Lyght, an Army Reservist, finds herself mutated from a military experiment gone wrong, givin...
Rapture Like Bloodby Noizchild (R)
Eighth one in my series, "Ghost Doll." Three cases in one: missing and moving corpses, Hotaru's frie...
In: Yami no Matsuei


text:
Monday, 15 Jun 2009 12:24 PM
By Dan Weil
The next shoe that looks ready to drop for the housing market is the adjustable rate mortgage sector.
ARMs, popular during the real estate boom, have low initial payments and then reset at loftier levels down the road.
About 1 million option ARMs will reset higher during the next four years, according to First American CoreLogic.
About 75 percent of those loans will shift in 2010 and 2011, with the zenith looming in August 2011, when about 54,000 loans reset, the data show.
The resets will pressure many homeowners, making their payments unaffordable.
"The option ARM recasts will drive up the foreclosure supply, undermining the recovery in the housing market," Susan Wachter, professor of real estate finance at the University of Pennsylvania's Wharton School, tells Bloomberg News.
"The option ARMs will be part of the reason that the path to recovery will be long and slow."
RealtyTrac reports that foreclosures totaled more than 321,000 in May, the third-highest monthly rate since the research firm began its report in January 2005, and the third straight month with more than 300,000 foreclosures.
To be sure, some found a bit of comfort in the fact that foreclosure filings fell 6 percent in May from April.
"It would not be a huge surprise to see the numbers level off a little bit at this point," Rick Sharga, RealtyTrac's senior vice president for marketing, told Associated Press.
© 2014 Newsmax. All rights reserved.
Powered by
How about we get some whistleblowers for "fast and furious" and have him answer...
The poor underpaid McD's Workers
They need $15/hr so they can get more tattoos,...
The minimum wage was established to give employers a way to hire students and...
We must get rid of "The Fed". We can start by indicting the Board members who...
Oh, please, simply abolish the privately-owned Federal Reserve, which is no...
Newsmax, Moneynews, and Independent. American. are registered trademarks of Newsmax Media, Inc. Newsmax TV, NewsmaxWorld, NewsmaxHealth, are trademarks of Newsmax Media, Inc.


text:
That's something to celebrate! Jennifer Aniston looks youthful in her blonde wig as she spends 44th birthday on movie set
She is one of Tinseltown's most celebrated beauties, and Jennifer Aniston showed her star power hasn't waned one bit as she stepped out on her 44th birthday.
The actress appeared years younger than some of her acting counterparts while working on the set of her new film on her special day.
Working on her special day: The star was on the set of the new untitled Elmore Leonard Film in Connecticut
Cold outside: Jennifer covered up in a padded coat and donned a brown dress and knee-high boots
Glowing skin: Jennifer was looking especially youthful on her 44th birthday
Jennifer was the picture of perfection as she stepped out wearing a fresh face of make-up on the set of the new untitled Elmore Leonard Film in Greenwich, Connecticut.
Apparently she wanted a low-key day as she carried out her lunch in a polystyrene box as she emerged from a door which read, 'Attention! No food pantry until Friday'.
She told PEOPLE magazine: 'I'll just be working. I know that I have a 5:30 am pick-up but that's not bad.
'I'm lucky, I'm very happy to be working!'
And like most people, she's wondering where the time goes.
Feeling happy: The star - who donned a blonde wig - has been announced for a new film project
Downcast: She paid little attendtion to photographers on set
'This birthday is going to come and go, which they all seem to be doing these days. Wasn't it just my birthday? I have like windburn from this year.'
But she expressed her idea of a perfect birthday 'would be a walk in the city, brunch somewhere, see a movie, have a great dinner.'
Jennifer is attached to the production for five weeks, and has relocated to the east coast. But she continues to fulfill her duties as spokesperson/co-owner for Living Proof hair care.
For her role in the film, she opted to wear a shorter, light blonde wig instead of coloring her famed tresses.
She said: 'It's so nice to wear a wig. It's like putting a hat on. I've been using our conditioners on my hair underneath and it's great to be able to watch your hair actually grow.'
She bangs! Jen was seen standing next to a clapped out car between takes
Jennifer is starring in an adaptation of Elmore Leonard novel, The Switch.
The story follows Ordell Robbie and Louis Gara, two career criminals in Detroit who team up to kidnap the wife of a corrupt real estate developer.
When their ransom demands backfire and the jilted housewife demands their assistance in seeking revenge, the two criminals must rethink their plan.
The former Friends star is set for a busy year, after it was announced she has signed on for motion picture She's Funny That Way.
Jennifer will be reunited with Marley & Me co-star Owen Wilson in the Peter Bogdanovich comedy in which she will play a therapist whose mother is in rehab for alcohol issues.
Retro look: Jennifer wore a burnt orange coat and tweed goucho pants on set
Emotional: The actress appeared focused on getting into character
Owen takes on the role of a married Broadway stage director who falls for a prostitute (Brie Larson).
When Brie's character shows interest in becoming an actress, he decides to help her with her career.
Other stars to have signed up for
the film - which has been co-written by Bogdanovich and Louise Stratton -
include Jason Schwartzman, Cybil Shepherd, Eugene Levy and Kathryn
Filming is set to start in May in New York.
Outfit change: She was later seen wearing a thick parka and dark sunglasses as she stayed warm with a coffee
Wet weather: Jennifer was sheltered underneath an umbrella as it poured down with rain
Earlier this month, it was revealed she had signed up for new romantic comedy Convention.
She is on board to play the female lead in Justin Reardon's 'mind-bending' feature film alongside Safety Not Guaranteed star Mark Duplass, which will see the dynamic duo revolutionise the box storage world thanks to the invention of a five-sided box.
And action! Jennifer with co-star John Hawkes
Warm embrace: The pair went in for a hug as they acted out a scene
In character: John seemed to smile while Jennifer was caught in the middle of an awkward facial expression
- Unknown creature seen in Chicago river believed to be a...
- RFK Jr loses his cool over his own carbon footprint
- F*** it I quit, news reporter quits live on air
- Ugly fight at the 49ers game vs Cardinals
- Obama salutes Marines with coffee cup in hand
- Tiger kills man who climbed into enclosure in New Delhi
- Adorable Budweiser ad encourages safe and responsible...
- 'F**k it, I quit' reporter explains why she resigned
- Don Spirit calls 911 after killing 6 grandkids and daughter
- Israeli military says it shot down Syrian war plane
- Leonardo DiCaprio and Jamie Foxx rapping up a storm
- Detroit Lions' Tulloch injures knee doing victory dance
- Tempest over Obama's tea cup salute: Commander-in-chief...
- Fake or freak? Woman, 21, claims to have paid $20,000 to...
- 'This is only the beginning': Pentagon reveal how dozens of...
- 'He tried to run us off the road!': Woman's terrified 911...
- What really happened behind Palace doors in royalty's...
- 'Drunk' man mauled to death by white tiger after leaping...
- BREAKING NEWS: New arrest warrant for abduction with intent...
- Robert F Kennedy Jr loses his cool and grabs mic from...
- They're fake! Woman who claims to have surgically implanted...
- Student 'raped teacher at high school after he body-slammed...
- 'We use MSG and we don't give a f*** about gluten-free': San...
- Cyclist whose 'speeding' crash killed CBS executive's wife...
The comments below have not been moderated.
The views expressed in the contents above are those of our users and do not necessarily reflect the views of MailOnline.
We are no longer accepting comments on this article.


text:
Phil Handler
Canyon Building fined $10,500 for asbestos violations
A Tucson construction company exposed workers to asbestos when it sent them to remove pipes from a job site because it didn't want another contractor to get the valuable copper, a state investigation found.
Follow the Arizona Daily Star
Featured businessesView more...
Deals, offers & eventsView more...
FREE Remote
with the purchase of a pair of Beltone Promise Hearing Aids
Our facility is available for Birthday Parties & Team Parties. Click here for more information!
SEPTEMBER SALE EVENT! Check out Quality Pottery Etc. for huge savings on all of the pottery pieces you love! Click here for sale details!
Have you seen all of our latest inventory? Now is the perfect …
10% OFF for a Location Check
10 % Off to locate your leak. Not in combination with any oth…
Whether you are moving across town or across the country, Ralph's Transfer Inc. will provide you with the peace of mind you deserve! (520) 622-6461
Whether you need local moving, in-state moving, cross country …
Event Calendar
Most viewed:


text:
Sunday, 24 Nov 2013 12:31 PM
By Audrey Hudson
Another key Democrat, Sen. Charles E. Schumer of New York, said in a statement that he was disappointed in the interim deal reached in Geneva regarding Iran's nuclear program, saying "it does not seem proportional" because "Iran simply freezes its nuclear capabilities while we reduce the sanctions."
The Senate agreed unanimously to keep financial sanctions on Iran, and the House passed a similar measure this summer on a 400-20 vote.
© 2014 Newsmax. All rights reserved.
Thursday, 02 Oct 2014 10:25 AM
Speaking out against recent Secret Service gaffes, at least one member of Congress is noting that it's a sad day when a . . .
Thursday, 02 Oct 2014 09:57 AM
As the White House considers a replacement for Attorney General Eric Holder following his resignation last week, insider . . .
Thursday, 02 Oct 2014 09:56 AM
Former Secret Service Director Julia Pierson was "famous" for saying that she wanted the agency in charge of the securit . . .
What this country needs is a hard working, adult President....
Your perpetual state of socialist bliss is possible due to the
Honestly, is anyone, ANYONE, at all surprised in the least that when being faced...
Ohhhhh but of course allow a man whose whole career has been hatred, race...
Simply indicative of the general ignorant attitudes that prevail about our...
Newsmax, Moneynews, and Independent. American. are registered trademarks of Newsmax Media, Inc. Newsmax TV, NewsmaxWorld, NewsmaxHealth, are trademarks of Newsmax Media, Inc.


text:
Jamie Bestwick 8-Peats in BMX vert
Jamie Bestwick talks about winning his 8th gold medal in BMX vert at X Games Barcelona 2013.
(01:40) | May 17, 2013
Latest Video & Audio More »
Play Video
Lochte glad to see Phelps back in action
More Sports Video: As he prepares to face Michael Phelps at the Pan Pacific Championships this week, Ryan Lochte says he's glad to see the 18-time Olympic champion back in action.
(01:52) | Aug 21, 2014
Play Video
50 Cent's 1st pitch fail
More Sports Video: Curtis Jackson, aka 50 Cent, threw out the ceremonial first pitch at Citi Field as the Mets hosted the Pirates. It may have been the first time he had thrown a baseball.
(00:19) | May 28, 2014
Play Video
Exclusive interview: Mark Selby
More Sports Video: 2014 snooker World Champion talks to Ismail Vedat about beating Ronnie O'Sullivan and his plans for fatherhood.
(07:30) | May 21, 2014
Latest Video & Audio More »
Play Video
Tevez left out Argentina squad
ESPN FC on TV: ESPN FC's Alejandro Moreno reacts to Tata Martino's decision to leave Carlos Tevez out of the Argentina squad despite his excellent form.
(00:57) | Sep 23, 2014
Play Video
Man United failing to control games
ESPN FC on TV: ESPN FC's Stewart Robson finds fault in Manchester United's inability to control matches.
(00:41) | Sep 23, 2014
Play Video
Teams not sitting back against Real Madrid
ESPN FC on TV: ESPN FC's Alejandro Moreno says teams in La Liga refuse to sit back and defend like their Premier League counterparts.
(01:23) | Sep 23, 2014
Play Video
Should Rodgers limit Gerrard's playing time?
ESPN FC on TV: The ESPN FC team believe that Steven Gerrard should not be playing three matches in a week for Liverpool.
(01:36) | Sep 23, 2014
Play Video
Burley: Man United deserved to lose
ESPN FC on TV: Former Chelsea midfielder Craig Burley isn't buying the argument that the referee was at fault for Manchester United's loss to Leicester City.
(01:35) | Sep 23, 2014
Play Video
Ronaldo on another scoring tear
ESPN FC on TV: Cristiano Ronaldo has scored 21 career hat-tricks in La Liga, one away from Alfredo di Stefano's record.
(00:36) | Sep 23, 2014


text:
Jiu Jitsu GearMMAWarehouse.com understands the significance of the Brazilian Jiu Jitsu/grappling community and the importance of Jiu Jitsu in MMA. All of our Brazilian Jiu Jitsu gear is designed and built to the high standards of professional grapplers in Brazilian Jiu Jitsu and MMA fighters who train in No-Gi and BJJ Gi. We seek to provide the best products specifically designed to improve and optimize performance so you can perform at your best.
Take a look at our bjj/grappling gear from the best brands in the world and equip yourself with excellent Jiu Jitsu and grappling gear from MMAWarehouse.com.
Showing 1 - 3 items
$89.99 - $99.99
Spend $50+ in Gracie and get a free sweat towel!
(2 ratings)
{boxId:"comparebdtvsiaai2BwgaaadgjnYmbhJU", id:"gra-1099", catId:"jiu-jitsu-gis"}
$88.99 - $94.99
Spend $100+ in Fuji and get a free Go Rin Elements Tee!
(1 ratings)
{boxId:"comparebdlfkiaai2zBkaaadgu9YmbhJU", id:"fuj-1049", catId:"jiu-jitsu-gis"}
(1 ratings)
{boxId:"comparebdyRIiaai2TecaaadgynYmbhJU", id:"gri-1022-0001", catId:"jiu-jitsu-gis"}


text:
These are all the News & Announcements posted on Monday, 1 October, 2007.
Banned Books Week
This week (Sept 29 - Oct 6) is Banned Books Week. Banned Books week is an annual event that celebrates freedom to read or the freedom to express one's opinion even if that opinion might be considered unorthodox or unpopular and stresses the importance of ensuring the availability of those unorthodox or unpopular viewpoints to all who wish to read them. To see a list of books that others have attempted to ban, view The American Library Association's list of The 100 Most Frequently Challenged Books.
Celebrate your freedom to read by selecting a book from a list of banned books available at the Logan Library.
Posted By Logan Library at 2:51 PM in Category:Books & Reading
Link directly to this article.
The Latest News!
Sep 18, 2014
» Hispanic Heritage Month / Mes de la Herencia Hispana Sep 15, 2014
» Send Us Your Book Selfie Sep 9, 2014
» Helicon West Showcases Local Authors


text:
Associated Press
RIGBY, Idaho —When she lost her diamond wedding ring four years ago, Necia Rasmussen held out hope for several months before finally buying a replacement. But she never stopped missing the one that got away.
She nearly fell over from joy last week when she was reunited with the platinum Princess cut diamond ring that washed up on an Idaho lake shore and was found by a kayaker four years after it went missing.
Rasmussen, 34, said she started crying when she claimed the ring after describing it in great detail and providing a photograph.
Rasmussen was at Rigby Lake near Idaho Falls four years ago when she took the ring off so she could rub sunscreen on her children. She placed the ring inside a chair's cup holder and thinks that it later fell out later when she was folding up the chair.
The loss was particularly upsetting because Rasmussen had recently learned that her husband, Heber, an Army reservist, was going to be deployed to Iraq. He had picked out the ring and it was platinum, just like his mother's wedding ring.
His mother also lost her wedding ring and it was found about three decades later, Rasmussen said.
"Thirty years later, his dad found it in the garden," she said.
In her case, she found herself reluctant to replace the ring that went missing at Rigby Lake, even with her husband's encouragement.
"I was having a hard time getting a new one," she said. "I just wanted my old one back."
The ring remained missing until Monday, when Emily Geisler spotted it while kayaking.
She said she hadn't been kayaking in a while and was practicing getting in and out of the watercraft on the shore when the ring caught her eye.
Geisler announced her finding at a meeting of the Rigby Rotary Club late Tuesday, after the group had finished its business. The announcement caught the attention of Rasmussen's brother-in-law, Glenn Walker, a fellow Rotary Club member.
"He happened to remember me and my sister talking about my ring missing a year ago," Rasmussen said.
Walker contacted his wife, who then contacted Rasmussen. He then approached Geisler, who remained skeptical until Walker started to describe the ring in specific detail, with Rasmussen on the other end of the phone telling him what her ring looked like.
Rasmussen then joined Walker at the restaurant where the Rotary Club was meeting. She provided a photograph of her ring, which was identical to the one in Geisler's possession. The ring was in good condition and just needed a cleaning. It fit Rasmussen perfectly.
"I totally believe in miracles," she said. "I had prayed about it, but I thought maybe somebody found it who needed it more than me."
- Mom battling cancer determined to live for...
- Ex-Tabiona High teacher sent to prison for...
- The ghosts under our feet: 88...
- Police break silence about controversial...
- 3 South Jordan siblings battling rare disease...
- Utah, Western states say feds are all wet on...
- 'Living nightmare' almost over for daughters...
- Police chief of Myton charged with stalking 3...
- 'They killed my son because he's... 79
- Jason Chaffetz: Mitt Romney is leaving... 67
- Was Saratoga Springs man killed while... 52
- Police break silence about... 44
- Friends, family, strangers gather at... 35
- Utah, Western states say feds are all... 20
- New definition of homeless would give... 20
- Utah's Gov. Gary Herbert eyes more... 11


text:
More Videos...
BMW Concept Cars
Film przedstawia prototypy BMW, zarówno te doskonale znane jak i te już
dawno zapomniane, zagubione w odmętach historii. Są w śród nich
projekty które miały bardzo duże szansę wejść do seryjnej produkcji,
jak i te których celem było ukazanie umiejętności projektantów i
zaprezentowanie możliwości firmy. Niektóre wciąż mają szansę ożyć!
First Video of 2016 BMW M2 F87 Prototype Testing on Nurburgring Track ! ByKayseri !
bugatti ace hood,bugatti Veyron,bugatti,Mustang,nissan gtr,lotus flower
bomb,audi r8,flying lotus,top gear,Camaro,bmw m3,ace hood
bugatti,bmw,toyota supra,corvette,dodge challenger,honda civic,corvette
zr1,Mustang cobra,Mustang sally,nissan skyline,lotus,Mustang gt,radiohead lotus
flower,dodge charger,Camaro ss,bmw m5,
cadillac records,honda,dark lotus,mercedes benz,dodge viper, audi,honda
civic turbo,honda s2000,bmw m6,2010 Camaro,lotus flower,turbo Mustang,Mustang 5.0,dodge cummins,sideways
dierks bentley,2011 Mustang
gt,corvette z06,audi a4,honda civic si,little red corvette,Camaro zl1,bentley green,audi
s5,nissan 350z,toyota,honda prelude,street racing,ford Mustang,honda accord,Mustang burnout,2010 Camaro ss, nissan
370z,bentley,lamborghini,supra,window seat,dierks bentley home,kia
soul,supra from hell,toyota tacoma,cadillac cts-v,supercharged Mustang,toyota tundra,dodge ram,audi
rs4,dodge dart,dodge,bugatti remix,Camaro vs Mustang,kia,zr1 corvette,Mustang vs Camaro,range rover,cadillac
kimberly,erykah badu,dierks bentley sideways,dirks bentley,future
bugatti,nissan honda chevy,audi a8,5.0 Mustang,audi tt,dodge tomahawk,bmw
x6,home dierks bentley
Supercharged BMW M3 E92 Amazing Drift & Sound - Francesco Conti' KOE Car
Flickr Page: http://www.flickr.com/photos/19bozzy92/
Facebook page: https://www.facebook.com/19Bozzy92YT
Google+ page: https://plus.google.com/100567781241722617164/posts
This video is about the fantastic supercharged BMW M3 E92 of Francesco
Conti, the champion of the 2013 King of Europe drifting series. Here you
can see him in action on a slightly wet track during the 4th round of the
2014 Kamikaze Drift Events.
Camcorder: Canon Legria HF M46 + Canon DM-100 Microphone
Event: Kamikaze Drift Events 2014 Round 4
Where: Castelletto di Branduzzo, Italy
Link To My Channel: http://www.youtube.com/user/19Bozzy92?feature=mhee
BMW M3 GTS VS BMW M5 F10 Ring Taxi Nürburgring Nordschleife with external mic + Harrys Laptimer
BMW M3 GTS VS BMW M5 F10 Ring Taxi Nürburgring Nordschleife 13 07 2014
► Subscribe here: http://goo.gl/IkIKxo
► Instagram: http://goo.gl/cVEQKG
► Like us: http://goo.gl/YAdLja
BMW 330(e46) Тест-драйв.Anton Avtoman.
http://vk.com/antonavtoman Добавляйтесь в друзья!)
Я в Инстаграме http://instagram.com/antonvorotnikov
BMW M3 CSL - Top Gear
On Top Gear 03x02 2003-11-02
2014 BMW M3 sedan is spied out in the Wild
Read all about the M3's full specs here: http://wp.me/p1sgG3-ZQ
Can you see it out there on the horizon? There is a car that is quickly
racing towards us. BMW may have two new M cars that are reaching their
final stages of development/testing and they look like they are keeping
their promise of class leading performance. Let us first explain the
reason for why two cars maybe in the works as the Boys from Bavarian are
creating the traditional M3 alongside a new two door version that is
rumored to be called the M4.
BMW M3 drift
Directed & Edited by Bahrom Jalilov
Post production by JALILOV.COM
Camera: EOS7D + 16-35 f/2.8
Dushanbe 2013
How To Drift, BMW M235i - /Chris Harris On Cars
Chris Harris explains how to drift with the new BMW M235i. See the full
BMW Drift Mob video here: https://www.youtube.com/watch?v=vz2rAgXjkCA
Rozbieracze BMW e36 M3 VS Mercedes 380 SL
Rozbieracze - Biznes w garażu (discovery) BMW e36 M3 VS Mercedes
The all-new BMW X6: All you need to know.
Have a look at the all-new BMW X6 and learn the most important facts in
less than 8 minutes. In this clip we compare the 2nd BMW X6 generation to
the first one, show you details of the new exterior and interior design,
listen to the sound and demonstrate the latest BMW ConnectedDrive features.
Register interest and get updates for the all-new BMW X6:
To see more clips about the all-new BMW X6 or to get the latest BMW clips
in general, just subscribe to the official BMW YouTube channel:
► The BMW i8 Production
BMW i8 deliveries to customers starting in June.
Plug-in hybrid system developed and produced by the BMW Group represents
the latest development stage of Efficient Dynamics; debut for
three-cylinder petrol engine with BMW TwinPower turbo technology, displacement: 1.5 litres,
output: 170 kW/231 hp, maximum torque: 320 Nm (236 lb-ft); power sent
to the rear wheels via a six-speed automatic gearbox; model-specific hybrid
synchronous electric motor, output: 96 kW/131 hp, maximum torque: 250 Nm
(184 lb-ft); power channelled through the front wheels via a two-stage
automatic transmission; lithium-ion high-voltage battery with liquid
cooling and usable capacity of 5 kWh.
Combination of BMW TwinPower turbo and
BMW eDrive technology plus intelligent energy management produce system
output of 266 kW/362 hp (max. torque: 570 Nm / 420 lb-ft) and give the
BMW i8 the performance characteristics of a pure-bred sports car (0 --
100 km/h / 62 mph in 4.4 seconds) combined with fuel economy and
emissions figures more familiar from a small car (EU fuel consumption:
2.5 litres per 100 km / 113 mpg imp); "glued-to-the-road" AWD driving
experience with torque distribution geared towards optimised dynamics.
Driving Experience Control switch and eDrive button allow driver to choose
from five driving modes; range of up to 35 kilometres (22 miles) on
electric power alone and a top speed of 120 km/h (75 mph); COMFORT mode
offers optimum balance between dynamics and efficiency; combined range in
everyday conditions: over 500 kilometres (310 miles); SPORT mode with
ultra-intense Boost function
provided by the electric motor; ECO PRO mode can be used in both
all-electric mode and hybrid mode.
Sophisticated chassis technology featuring a double-wishbone front axle and
a five-link rear axle; Electric Power Steering; Dynamic Damper Control
comes as standard; 20-inch light-alloy wheels are standard, carbon wheels
are optional.
Emotion-led visual impression based around established BMW i design
language; classical sports car proportions and fresh interpretation of BMW
design features; doors open upwards like wings; clean lines, plus surface
design (external and internal) based on the layering principle; full-LED
headlights as standard, innovative laser headlights -- unique worldwide --
available as an option.
Intelligent lightweight construction with elements including a CFRP
passenger cell, doors with a CFRP-aluminium structure, an instrument panel
with magnesium support, an aluminium chassis and a partition between the
passenger compartment and boot made from thin glass; comprehensive safety
concept and an ultra-torsionally stiff passenger cell.
Extensive standard equipment includes the Navigation system Professional
with proactive drive system for all-electric driving, fully-digital
instrument display, BMW iDrive with freestanding Control Display and
leather sports seats; choice of four exterior paint finishes and four
interior equipment variants.
Wide range of BMW ConnectedDrive features: Park Distance Control, cruise
control system with braking function, rain sensor and Intelligent Emergency
Call function as standard; optional driver assistance package with High
Beam Assistant, a rear view camera, Surround View, Speed Limit Info
including No Passing Info display, and Collision Warning with pedestrian
recognition and braking function; also available are the Head-Up Display,
BMW Online Entertainment, Concierge Services, Real Time Traffic
Information and mobility services developed specifically for BMW i, e.g.
intermodal route guidance.
► SUBSCRIBE NOW: http://urlz.fr/lEd
► WEBSITE: http://www.youcarnews.com
► FACEBOOK: http://www.facebook.com/youcarpress
► TWITTER: http://www.twitter.com/youcartv
► GOOGLE+: http://plus.google.com/+YouCarPress
EXCLUSIVE: First Review Of The All New BMW M235i. Does It Resurrect The Soul Of The E36 M3?
Automotive social media news site AutoSpies.com (The TMZ/WikiLeaks of
autos) breaks this exclusive video review of the all new BMW M235i and a
showcase of the M-Performance package add ons available for it. Is it a
worthy successor to the 1m? Should the competition fear it? Is it a
resurrected E36 M3 geared towards the BMW traditionalists? To find out plus
more info including hundreds of photos and more details of the vehicle
visit the number one automotive social media news site www.AutoSpies.com.
Petrol BMW M3 vs diesel Alpina D3 - fast saloon showdown
Subscribe to the channel now: http://smarturl.it/autocar
Steve Sutcliffe has the keys to two very fast versions of the BMW 3-series:
the 435bhp petrol BMW M3 and the 345bhp diesel Alpina D3. Should you
satisfy your high-performance saloon cravings with a petrol or a diesel
engine? Let's find out.
Autocar, the world's leading motoring magazine and website, delivers
industry-leading news, the most in-depth car reviews and opinion from our
team of experts. Our presenters include some of the world's top motoring
journalists who have unrivalled access to the world's fastest, rarest, most
exotic and most exciting cars on some of the world's best roads and race
Visit http://www.autocar.co.uk for the latest news, reviews and analysis
New videos are uploaded to Autocar's YouTube channel each week. Don't miss
a single one. Subscribe to the channel now: http://smarturl.it/autocar
LIKE Autocar on Facebook: http://www.autocar.co.uk/facebook
FOLLOW Autocar on Twitter: https://twitter.com/autocar
+1 Autocar on Google+: https://plus.google.com/+autocar
2015 BMW M3: Restoring the Faith, BMW M is Back! - Ignition Ep. 112
On this episode of Ignition, Carlos Lago heads to a rain soaked Road
America to test the limits of the all new BMW M3! Reworked from the ground
up, the 2015 M3 ditches the screaming V8 of the last generation for an all
new twin turbocharged inline six
producing 425hp and 406lb-ft of twisting force. Designed to outperform its
predecessor in every measurable way, has the M3 become numb in its quest
for higher performance, or will it deliver on its promise of being the best
driving M yet? Stay tuned to find out!
Ignition appears every other Monday on the Motor Trend youtube channel.
Subscribe now to make sure you're in on all the action!
Facebook - http://facebook.com/motortrendmag &
Twitter - http://twitter.com/motortrend & https://twitter.com/mt_ignition
Google+ - https://plus.google.com/+motortrend/posts
Instagram - @mt_ignition
Website - http://www.motortrend.com
Lamborghini Huracan Driving Event and 60-130 MPH Testing
Ford GT McLaren MP4-12C Lamborghini Gallardo Super Trofeo Stradale Photo Gallery
GoPro Winner for August 2014 is Steve with a 2002 Dodge Dakota
Lamborghini Huracan vs Porsche 911 Turbo S PDK Racing
Which car is faster?
Similar 1/4 mile timeslips to browse:
1994 Fiat Coupe t16: 9.405 @ 161.860
Lanno Angelo, Engine: B&L engineering, Supercharger: no Turbos: Garrett gt 4202r Tires: hoosier 275/45/17 d.o.t.
1990 Audi Coupe Quattro: 9.780 @ 145.980
Aaron C, Engine: 20v 2.0L Integrated Engineering stroker 4cyl, Turbos: Borg Warner S400sx 67mm Billet divided T4 1.25 A/R Tires: 4 X M&H Racemaster 8.5/24.5-15 Drag Slicks
1990 Audi Coupe Quattro: 9.907 @ 141.440
Aaron, Engine: Integrated Engineering 2.0LStroker, Turbos: Bullseye S366XL T4 divided 1.10 A/R @ 32psi Tires: 4 X M&H Racemaster 8.5/24.5-15 Drag Slicks
1995 Fiat Coupe 2.0 t16 4x4: 10.277 @ 137.270
Magrelli Marco, Engine: fiat 1995cc, Turbos: garrett gt3582r Tires: bfgoodrich t/a drag radial 205/50R15
1995 Fiat Coupe 2.0 t16 4x4 drag: 10.544 @ 133.980
Marco Magrelli, Engine: 1995 fiat, Turbos: gt3582r Tires: toyo r888 195/50/16
1990 Audi Coupe 20vt QUATTRO: 10.706 @ 135.660
janis Krauklis, Engine: 7A, Turbos: gt35r Tires: M/T drag radials
1995 Fiat Coupe 2.0 16v turbo 4x4: 10.747 @ 130.850
magrelli marco, Turbos: t3/60-1 Tires: 195/50r16
1995 Fiat Coupe 2.0 16v turbo 4x4: 10.747 @ 130.850
Magrelli Marco (wind), Turbos: full race garrett T3/60-1 Tires: 195/50zr16 toyo r888
1990 Audi Coupe quattro 20vt: 10.809 @ 129.050
janis, Engine: 7a, Turbos: GT35r Tires: MT 225/50/15 drag radials
1995 Audi Coupe S2: 10.970 @ 128.000
Audrius, Engine: 2.2t 20v, Turbos: Holset hx40 Tires: yokohama 048m
1991 Audi Coupe S2: 11.477 @ 124.650
Zeki Fidan, Engine: 2226ccm, Supercharger: no Turbos: kkk 29 Tires: 234/35/17
2000 Buell M2 : 11.731 @ 112.950
Matt Bluett, Engine: 1203cc, Tires: stock
2000 Buell M2 : 11.794 @ 112.630
Matt Bluett, Engine: 1203cc, Tires: stock
2006 Superformance Coupe Brock: 11.900 @ 119.000
NA, Engine: Stock Roush 427ci V8, Tires: Kumho Ecsta V710: inflated to 24psi Rear: 315/35R1
1929 Ford Coupe : 12.000 @ 111.000
JB, Engine: Chevy 400, Supercharger: 4-71 Turbos: 400 Tires: Firestone/McQuiry
2003 Maserati Coupe Cambiocorsa: 13.000 @ 110.410
2004 Maserati Coupe Cambiocorsa: 13.300 @ 109.000
1997 Fiat Coupe 20 VT: 13.842 @ 0.000
Alen Pirgo, Turbos: GT 28R
1999 Fiat Coupe : 13.900 @ 100.650
Dab, Engine: Stock bar ECU remap, Turbos: stock
©2014 DragTimes - Disclaimer


text:
The official site of strivectin anti-aging products. free, The official site of strivectin anti aging products offers multiple creams, scrubs & serums with wrinkle reducing, skin tightening & moisturizing benefits..Game tracker - server stats, list search, ranking, banners, Track your server's world ranking see detailed daily usage stats collect top player statistics get detailed server banners. You., Share your videos with friends, family, and the world.
Wacky mania - just another wordpress weblog, Mirrors of memories. here you will find a unique photo project made by famous photographer tom hussey, he is very talented, his work wasled 'mirrors' it's.Stevelange.net › making your own minecraft steve head, As with most parents, i have an 11yr old that is mad mad mad on minecraft!! he is going to his disco its fancy dress, so it was "minecraft steve" here i. Wharton's spike® - university of pennsylvania, Wharton's spike, a suite of web-based applications and services, gives students simple and integrated access to course materials, resources, information, and online.
The best free video maker for you- ezvid.com, Fast & easy screen recorder. instant facecam & voice synthesis. awesome video editor. fun to use. instant slideshow maker. beautiful screen capture for games, desktop.


text:
Paul Laurence Dunbar
From The Porch At Runnymede
I stand above the city's rush and din,
And gaze far down with calm and undimmed eyes,
To where the misty smoke wreath grey and dim
Above the myriad roofs and spires rise;
Still is my heart and vacant is my breath--
This lovely view is breath and life to me,
Why I could charm the icy soul of death
With such a sight as this I stand and see.
I hear no sound of labor's din or stir,
I feel no weight of worldly cares or fears,
Sweet song of birds, of wings the soothing whirr,
These sounds alone assail my listening ears.
Unwhipt of conscience here I stand alone,
The breezes humbly kiss my garment's hem;
I am a king--the whole world is my throne,
The blue grey sky my royal diadem.
Paul Laurence Dunbar's Other Poems
Read this poem in other languages
This poem has not been translated into any other language yet.
I would like to translate this poem »
Comments about this poem (From The Porch At Runnymede by Paul Laurence Dunbar )
PoemHunter.com Updates
Biography of
Dante Gabriel Rossetti
Brass Spittoons
by Langston Hughes
Best quotes of
F. Scott Fitzgerald
Read more about
Anne Sexton
New Poems
- LET PRESIDENT UHURU KENYATTA COMPLY WITH.., alexander opicho
- Khak me mil jaye ya..., Azhar Sabri
- Coding By Instruments, RoseAnn V. Shawiak
- Coming Out, Edward Kofi Louis
- Teachers Of Peace, Edward Kofi Louis
- Under Tree Tops, RoseAnn V. Shawiak
- A Friendship Falling Into Love!, Kamini Arichandran
- In Your Neighbourhood, Edward Kofi Louis
- COPLA 114 CONCLUSION: This Bad Guy World, T (no first name) Wignesan
- Heal The World With Love, Edward Kofi Louis
Poem of the Day
Trending Poems
- 04 Tongues Made Of Glass, Shaun Shane
- Still I Rise, Maya Angelou
- Daffodils, William Wordsworth
- The Road Not Taken, Robert Frost
- If You Forget Me, Pablo Neruda
- I Know Why The Caged Bird Sings, Maya Angelou
- Phenomenal Woman, Maya Angelou
- Autumn Song, Dante Gabriel Rossetti
- O Captain! My Captain!, Walt Whitman
- If, Rudyard Kipling
Trending Poets
Shaun Shane
(196? -)
Maya Angelou
(4 April 1928 - 28 May 2014)
Jose Rizal
Robert Frost
(March 26, 1874 – January 29, 1963)
Wilfred Owen
- Heather Burns
Pablo Neruda
(12 July 1904 – 23 September 1973)
William Wordsworth
Edward Thomas
(3 March 1878 - 9 April 1917)
Charles Bukowski
(16 August 1920 – 9 March 1994)