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Hepatocyte growth factor (HGF) has been shown to enhance recovery from renal tubular ischemia. We investigated the possibility that HGF improves recovery by preventing ischemia-induced loss of cell adhesion. Murine inner medullary collecting duct-3 (mIMCD-3) cells subjected to 90% ATP depletion demonstrated a 55% decrease in adhesion, an effect that was completely reversed by the addition of HGF. Assays examining release of adherent cells revealed similar results with 30 min of ATP depletion causing loss of adhesion of 25% of mIMCD-3 cells and HGF completely reversing this effect. In contrast, HGF was unable to reverse the loss of adhesion of cells exposed to 99% ATP depletion. Examination of the mitogen-activated protein kinase (MAPK) signaling pathway revealed that HGF could induce extracellular signal-regulated kinase (ERK) phosphorylation in control and 90% ATP-depleted cells but not in 99% ATP-depleted cells. Inhibition of ERK activation with U0126 completely blocked the HGF-dependent reversal of ATP-depleted cell adhesion. Thus ATP-depleted cells demonstrate a marked decrease in cell adhesion that is reversible by the addition of HGF. This effect of HGF requires activation of the MAPK pathway.
- extracellular signal-regulated kinase
- renal ischemia
- inner medullary collecting duct
- mitogen-activated protein kinase
- adenosine 5′-triphosphate
hepatocyte growth factor(HGF) is a circulating protein that is produced primarily by fibroblasts and peritubular interstitial cells in a variety of organs including liver, kidney, lung, and pancreas (8, 31, 46,52). The addition of HGF to cultured epithelial cells may result in at least three distinct responses: mitogenesis, motogenesis (scattering/chemotaxis), and morphogenesis (branching tubule formation) (5). These responses are mediated by the c-met tyrosine kinase receptor and are likely to be involved in developmental and repairative epithelial cell migration and branching tubule formation (8, 18, 44, 46).
There is significant evidence to suggest that HGF and its receptor c-met play an important role in determining tubular cell fate after acute tubular injury. In the adult rat kidney, several laboratories have found that transient injury results in rapid upregulation of both HGF (at multiple sites) (16, 17) and c-met (selectively at the site of injury) (18, 25), suggesting a role for this growth factor receptor in tubular repair. Similarly, HGF levels were increased after tubular injury with either CCl4 or HgCl2 (21, 30). Two studies have shown accelerated recovery from models of acute tubular necrosis in rodents after the administration of HGF. In a study by Miller and colleagues (29) examining renal artery cross-clamp in the rat, the administration of HGF 30 min after reperfusion decreased the peak rise of blood urea nitrogen (BUN) and creatinine and hastened the recovery back to baseline renal function (29). By using exposure to either HgCl2 or cisplatin in mice to cause acute tubular injury, Kawaida and co-workers (21) found that pretreatment with HGF prevented the doubling of BUN that occurred in control mice at 24 and 48 h after toxin exposure. In both studies, histology revealed that control animals had widespread cell necrosis and areas of denudation of the tubular basement membrane, whereas HGF-treated animals had almost no areas of tubular cell loss. Thus HGF appears to not only enhance recovery but can actually diminish the acute injury.
After renal ischemia, the initial response of the tubular epithelial cells is loss of both polarity and tight junctional integrity followed over hours to days by one of three cell outcomes: frank necrosis, detachment and apoptosis, or dedifferentiation into a more fibroblast phenotype (thinning and spreading of the cytoplasm to cover the denuded basement membrane) (2, 47). Many of the cells that detach after acute renal injury are actually viable and can be cultured from urine (12, 26,35-37). These cells, instead of dedifferentiating and serving to aid in repopulation and recovery, are a contributor to the luminal obstruction that occurs after acute injury and therefore actually serve to worsen acute renal failure (23, 47). The mechanism by which otherwise viable cells undergo detachment from the basement membrane, and strategies to prevent this, are therefore of great interest. We investigated the possibility that part of the mechanism by which HGF can minimize renal tubular injury and promote repair is by enhancing cell adhesion of ischemic, ATP-depleted epithelial cells.
In an attempt to mimic the effects of in vivo ischemia on cell adhesion, we used a combination of the mitochondrial inhibitor oligomycin A and the glycolytic inhibitor 2-deoxyglucose to induce ATP depletion in cultured murine inner medullary collecting duct-3 (mIMCD-3) cells (28, 43). Conditions were determined whereby cultured renal epithelial cells are substantially ATP depleted for 30 min but recover if ATP levels are restored. We found that ATP-depleted cells failed to adhere to culture plates and that this diminished adhesion was reversed by treatment with HGF. In addition, we found that 25% of adherent cells would release attachment in the setting of ATP depletion, an effect that was again reversed by treatment with HGF. This response is fully prevented by the mitogen-activated protein kinase (MAPK) kinase (MEK) inhibitor U0126, suggesting that HGF-mediated cell adhesion is dependent on activation of the MAPK-signaling pathway.
Cells and reagents.
Experiments were performed with mIMCD-3 cells, immortalized murine inner medullary collecting duct cells of ureteric bud origin (39) that have been shown to express the c-met receptor and undergo migration and tubulogenesis in response to HGF (5). All cells were grown in DMEM-Ham's F-12 medium (DMEM/F-12) media supplemented with 10% FCS in a 5% CO2environment. HGF, 2-deoxy-d-glucose, oligomycin A, and poly-l-lysine were obtained from Sigma. Laminin, fibronectin, and collagen type IV were obtained from Becton Dickinson. Type I collagen was obtained from Upstate Biotechnology, and U0126 was obtained from Promega.
ATP depletion model.
mIMCD-3 cells were bathed in human peripheral-type benzodiazepine receptor (HPBR) buffer solution [(in mM) 135 NaCl, 5 KCl, 3.33 NaH2PO4, 0.83 Na2HPO4, 1 CaCl2 , 1 MgCl2, and 5 HEPES, pH 7.4] for 20 min and then exposed to HPBR containing either 10 mM 2-deoxyglucose and 5 μM oligomycin A or 100 μM 2-deoxyglucose and 25 nM oligomycin A for 30 min (3, 28, 50). For recovery experiments, the cells were washed once with HPBR buffer, and the media were replaced with standard DMEM/F-12 media containing 17.5 mM glucose. ATP levels were determined using the Bioluminescence Detection Kit for ATP (Promega). Briefly, mIMCD-3 cells were extracted with 2.5% trichloroacetic acid, the sample was neutralized with 250 mM Tris-acetate (pH 7.75), and ATP levels were measured using the Luciferase/Luciferin (L/L) Regent according to the manufacturer's protocol. A 2-s delay time after L/L Reagent injection and a 10-s relative light unit signal integration time were used. An ATP standard curve was obtained by adding 10 μl of ATP standard diluted to the proper concentrations and assayed using L/L Regent.
Cell adhesion assay.
To examine cell adhesion on different matrix substrates, plastic tissue culture dishes were coated overnight at 4°C with either laminin (1 μg/ml in DMEM/F-12), type I collagen (1 μg/ml in PBS), type IV collagen (1 μg/ml in 0.05 N HCl), fibronectin (1 μg/ml in DMEM/F-12), or polylysine (0.01% in water) as per manufacturer's instructions. mIMCD-3 cells were harvested by bathing in 1 mM EDTA for 10 min. The cells were then counted, and 4 × 104cells/well were plated on the coated 96-well plates in DMEM/F-12 media for the indicated time. Nonadherent cells were removed by being washed once with PBS, and remaining cells were quantitated by the method of Oliver et al. (34). Briefly, cells were fixed for 30 min with 10% formalin, stained for 30 min with 1% methylene blue, and washed three times with 0.01 M boric acid (pH 8.5). The intracellular dye was extracted with 0.1 N HCl/ethanol (1:1) for 1 h at room temperature and quantified by absorbance at 655 nm.
To examine adhesion after ATP-depletion, cells were harvested with 0.5% trypsin, 5.3 mM EDTA for 5 min. Equal numbers of suspended cells were incubated in either control media (DMEM/F-12) or HPBR buffer for 20 min. Trypsin had to be utilized to harvest cells for these experiments to prevent excessive cell clumping during the incubation period. Appropriate concentrations of 2-deoxyglucose and oligomycin A were then added to the cells suspended in HPBR for an additional 30 min. The cells were then pelleted, resuspended in DMEM/F-12 ± HGF (40 ng/ml), and plated for 30 min on a 96-well plate. For inhibitor studies, 20 μM U0126 was added at the same time as the HGF. Nonadherent cells were removed by washing once with PBS, and adherent cells were quantified, as above. To compare multiple experiments, adhesion of control cells was designated as 100%, and all other values were normalized to the control value. Each individual well representsn = 1, and each experiment was repeated on at least two separate occasions. P values were obtained using the Student's t-test.
Cell release assay.
mIMCD-3 cells were harvested by bathing in 1 mM EDTA for 10 min, counted, and 4 × 104 cells/well were plated on 96-well plates in DMEM/F-12 media and allowed to adhere for 2 h. Nonadherent cells were removed by being washed once with PBS, and the media were changed to control media (DMEM/F-12) or HPBR buffer for 20 min. Cells in the HPBR buffer were then ATP depleted for 30 min by the addition of 2-deoxyglucose and oligomycin A ± HGF. Cells that had released from the plate after the 30-min incubation were removed by washing once with PBS, and remaining cells were fixed and quantitated, as described above.
Western protein analysis.
Adherent mIMCD-3 cells were serum starved overnight followed by incubation in control media or ATP depletion media for 30 min, as described. In the MEK inhibition experiments, 20 μM U0126 or vehicle control was added for 20 min during the 30-min incubation in ATP depletion buffer. The cells were then stimulated with 40 ng/ml HGF or vehicle control for 10 min, lysed with radioimmunoprecipitation assay lysis buffer [(in mM) 160 NaCl, 20 Trizma base, 1 EDTA, 1 EGTA, 1 dithiothreitol, 1% triton X-100, 1% sodium deoxycholate, 0.1% sodium dodecylsulfate, 1 sodium orthovanadate, 1 sodium fluoride, 1 phenylmethylsulfonyl fluoride, 0.5 μg/ml leupeptin, and 0.5 μg/ml pepstatin A], and insoluble debris removed by centrifugation. Forty micrograms of cell lysates were then separated on 10% SDS-PAGE and immunoblotted with either anti-phospho MAPK or anti-extracellular signal-regulated kinase (ERK) (New England Biolabs).
HGF increases renal epithelial cell adhesion.
Initial experiments were designed to determine whether HGF can enhance adhesion of control mIMCD-3 cells. A time course of adhesion revealed that HGF had no effect on initial cell attachment at 5 min but caused a more rapid increase in adhesion at 30 and 60 min (Fig.1). By 2 h, all of the cells were adherent, with no difference between HGF-treated and control cells. We also compared adhesion at 30 min to commercial plastic tissue culture dishes coated with laminin, type I collagen, type IV collagen, or fibronectin vs. cell adhesion to poly-l-lysine-coated dishes. Consistent with the results seen in carcinoma cells (15), treatment with HGF increased mIMCD-3 cell adhesion to plastic dishes, type I collagen, and fibronectin (data not shown). In contrast, HGF had no effect on attachment to dishes coated with laminin, type IV collagen, or poly-l-lysine in these cells.
Reversible ATP depletion in mIMCD-3 cells.
In an attempt to mimic in vivo transient ischemia, we utilized the combination of the mitochondrial inhibitor oligomycin A and the glycolytic inhibitor 2-deoxyglucose to induce ATP depletion. mIMCD-3 cells exposed to 25 nM oligomycin A and 100 μM 2-deoxyglucose for 30 min demonstrated an ∼90% decrease in intracellular ATP concentration (Fig. 2 A), whereas ATP levels in cells treated with 5 μM oligomycin A and 10 mM 2-deoxyglucose for the same time period declined to 1% of baseline values (Fig.2 B). Of note, treatment with HGF did not alter the degree of ATP depletion induced by oligomycin A and 2-deoxyglucose (90% ATP depletion = 9.3 ± 0.5% of control ATP levels, 90% ATP depletion + HGF = 11.0 ± 0.58%; 99% ATP depletion = 1.82 ± 0.1%, 99% ATP depletion + HGF = 2.02 ± 0.1%, n = 3 for all conditions). When the oligomycin A and 2-deoxyglucose were removed and cells were cultured in glucose-containing media, ATP levels fully recovered within 1 h in the 25 nM oligomycin A/100 μM 2-deoxyglucose-treated (90% ATP-depleted) cells but recovered to only 20% of baseline values in the 5 μM oligomycin A/10 mM 2-deoxyglucose-treated (99% ATP-depleted) cells. After 30 min of ATP depletion, 1.3% of 90% ATP-depleted cells and 2.3% of 99% ATP-depleted cells were judged to be dead by trypan blue staining. When these ATP-depleted cells were allowed to recover for 2 h suspended in normal media, 1.7% of 90% ATP-depleted cells and 2.7% of 99% ATP-depleted cells were trypan blue positive. In contrast, 5.3% of cells maintained for 2 h in 90% ATP depletion media were trypan blue positive and 34% of cells maintained for 2 h in 99% ATP-depletion media were judged to be dead. Therefore, 30 min of ATP depletion results in cells that appear to remain viable if resuspended in normal, glucose-containing media.
ATP depletion causes reversible loss of cell adhesion.
To examine the effects of ATP deletion on cell adhesion, suspended mIMCD-3 cells were subjected to 30 min of ATP depletion media or control media before adhesion for 30 min on 96-well plates. Ninety percent ATP depletion resulted in a 55% decrease in mIMCD-3 cell adhesion compared with control cells, whereas 99% ATP depletion resulted in <1% of the cells adhering after 30 min (Fig.3 A). The addition of HGF at the time of plating again resulted in increased adhesion of control cells and completely reversed the loss of adhesion seen in 90% ATP-depleted cells. In contrast, HGF treatment of mIMCD-3 cells subjected to 99% ATP depletion resulted in only a modest increase in the number of cells adhering to the plate.
In an attempt to mimic the conditions of cell detachment after ischemic acute renal failure, we examined the effect of ATP depletion on detachment of adherent cells. mIMCD-3 cells that were plated and allowed to fully adhere (2 h) were subjected to ATP depletion for 30 min followed by the removal of detached cells and quantitation of remaining cells. As has been described in the renal tubule after transient ischemia, 90% ATP depletion resulted in the detachment of 25% of the cells at 30 min, whereas 99% ATP depletion causes ∼40% of the cells to lose attachment (Fig.3 B). Of note, in agreement with the in vivo results of Racusen (35), 45% of the cells that detached after 30 min of 90% ATP depletion reattached and proliferated when replated in normal culture media, demonstrating that these cells were viable at the time of detachment (data not shown). Similar to the effects seen in the ATP-depleted cell adhesion assay, the addition of HGF at the time of ATP depletion completely prevented the release of mIMCD-3 cells subjected to 90% ATP depletion. In 99% ATP-depleted cells, there was no significant effect of HGF on cell detachment.
We then examined the effects of ATP depletion on mIMCD-3 cell morphology. When these cells are plated on plastic dishes, they are initially rounded. In the absence of ATP depletion, over several hours the cells extend processes in multiple directions and flatten out on the plate (Fig. 4, top). In contrast, when cells were either 90% ATP depleted or 99% ATP depleted before plating, the process of flattening and of extension was severely inhibited (Fig. 4, middle left and bottom left). Treatment with HGF at the time of plating had no further effect on spreading of control cells at 2.5 h (top right) but dramatically reversed the loss of process extension and flattening observed in 90% ATP-depleted cells (middle right). In 99% ATP-depleted cells, HGF failed to reverse the loss of process extension and flattening in the majority of cells (bottom right).
The HGF-mediated increase in cell adhesion is MAPK dependent.
HGF stimulation of mIMCD-3 cells results in activation of multiple signaling pathways including the MAPK proteins ERK1 and ERK2 (9,13). We have demonstrated that activated ERK2 can bind and phosphorylate the membrane-associated protein GAB1 after HGF stimulation (41), and active ERK has been recently demonstrated to associate with focal adhesions at the sites of cell attachment after src activation (11). These results suggest that activation of ERK can result in its recruitment to focal adhesion complexes where it might phosphorylate proteins that are critical for cell adhesion. To determine whether activation of ERK is critical for the observed HGF effects on ATP-depleted cell adhesion, we first investigated whether HGF can stimulate ERK activation in ATP-depleted cells. Adherent mIMCD-3 cells were pretreated with control media or ATP depletion media ± the MEK inhibitor U0126 for 30 min, followed by 10 min of stimulation with HGF. Whole cell lysates were then probed with an antibody directed against activated ERK1/2. The addition of HGF to control mIMCD-3 cells resulted in phosphorylation and activation of ERK1 and 2 as has been previously described (Fig. 5 A). This phosphorylation of ERK after HGF stimulation still occurs in 90% ATP-depleted cells but fails to occur in 99% ATP-depleted cells. Pretreatment with the MEK inhibitor U0126 completely prevents ERK activation. Reprobing of this same blot with an antibody to total ERK1/2 reveals essentially equal loading of protein in all lanes.
To determine the importance of HGF-induced ERK activation for the increase in ATP-depleted cell adhesion, we examined the effects of the MEK inhibitor U0126 on adhesion of mIMCD-3 cells plated for 30 min. A dose-response curve demonstrated that 10 μM U0126 was required for complete inhibition of HGF-induced ERK1 and ERK2 activation (data not shown). Pretreatment with 20 μM U0126 prevented the baseline increase in adhesion seen in control cells after HGF stimulation (Fig.5 B) and completely reversed the HGF-dependent increase in adhesion of 90% ATP-depleted cells. Thus the increase in mIMCD-3 cell adhesion after HGF treatment requires activation of the ERK-signaling pathway.
Acute renal failure, whether caused by ischemic or toxic injury, can result in exfoliated renal tubular cells that are morphologically intact and still viable, ultimately resulting in apoptosis of these cells (35). HGF and c-met mRNA levels increase significantly in animals after acute ischemic or toxic injury (16, 18), and the administration of exogenous HGF diminishes tubular injury and accelerates renal regeneration in animal models of acute renal failure (21,29). It has recently been demonstrated that carcinoma cells stimulated with HGF will more rapidly adhere to either plastic dishes or dishes coated with various extracellular matrix components, including type 1 collagen, laminin, and fibronectin (1,48), suggesting that the HGF/c-met signaling system can regulate cell-matrix adhesion. We therefore examined the possibility that HGF can regulate adhesion in normal and ischemic renal tubular epithelial cells.
Cell adhesion is a complex, multistage process in which suspended, rounded cells initially engage matrix and form focal contacts via a divalent cation-dependent interaction with cell surface proteins known as integrins (14). The rate of engagement of the integrin with matrix is dependent on the affinity state of the extracellular domain of the integrin, a process that is not fully understood but believed to be dependent on the interaction of the cytoplasmic domain with integrin-binding proteins such as talin, α-actinin, and focal adhesion kinase (FAK). Presumably, a conformational change in the extracellular domain of the integrin then allows binding to the matrix ligand, such as the classic RGD motif in fibronectin (19). The focal contact formed by the αβ-integrin heterodimer binding to matrix matures to form a focal adhesion, and clusters of these focal adhesion complexes become major sites of actin-filament attachment and trigger the process of cell spreading. Cell spreading provides more surface area for focal contacts, resulting in progressive focal adhesion complex formation and clustering and eventually a tightly adherent cell.
The mechanism by which HGF induces increased adhesion is not well understood. In the present experiments, HGF had no effect on adhesion to poly-l-lysine [a positively charged substrate that mediates cell attachment via charge interactions with anionic proteins on the cell surface rather than by integrin engagement (40,45)] but did increase adhesion to the integrin-specific substrates fibronectin and type 1 collagen. This result suggests that HGF induces adhesion in an integrin-dependent manner in renal epithelial cells and is in agreement with two studies of carcinoma cells that have found that increased adhesion to laminin after HGF stimulation was prevented by neutralizing antibodies to α2/β1-, α3/β1-, and α6/β4-integrins (1, 48). Because HGF is known to cause "scattering" (32), a process in which epithelial cells flatten, lose their cell-cell contacts and migrate away from each other, it is possible that the effect of HGF on cell adhesion is mediated by more rapid flattening and spreading of the cell, allowing more surface area for contact with the substrate and more rapid formation of focal adhesion complexes. In agreement with this, a time course of adhesion reveals that HGF has no effect on initial attachment of mIMCD-3 cells at 5 min (suggesting that HGF does not alter the affinity state of unengaged integrins) but does increase adhesion at 30 and 60 min (consistent with an effect on focal adhesion maturation and cell spreading after initial attachment).
In an attempt to determine the potential effects of in vivo ischemia on cell adhesion, we utilized the classic in vitro model of ATP depletion. By removing cells from glucose and exposing them to the combination of the mitochondrial inhibitor oligomycin A and the glycolytic inhibitor 2-deoxyglucose, graded intracellular ATP depletion can be achieved (6, 28, 42, 43). In an effort to distinguish sublethal and lethal injury, conditions were defined whereby cells are substantially ATP depleted for 30 min but recover if ATP levels are restored. Based on the results of Venkatachalam et al. (49) and Lieberthal and coworkers (24) in proximal tubular cells, we predicted that transient 90–99% ATP depletion would be sublethal, whereas sustained 99–100% ATP depletion should be lethal to our renal tubular cells. In agreement with this, mIMCD-3 cells exposed to 30 min of 25 nM oligomycin and 100 μM 2-deoxyglucose showed ATP levels between 10 and 13% of baseline and were fully recovered when glucose was added for 1 h, whereas ATP levels in cells treated with 5 μM oligomycin and 10 mM 2-deoxyglucose for the same time declined to 1% of baseline values and recovered only to 20% of baseline values over the ensuing 60 min. Despite this slow recovery of ATP levels in the 99% ATP-depleted cells, cell survival (as judged by trypan blue exclusion) was not significantly different in transiently 90% ATP-depleted and 99% ATP-depleted cells (1.7 ± 0.6 dead and 2.7 ± 0.6% dead, respectively). In contrast, sustained 99% ATP depletion for 2 h resulted in >30% trypan blue-positive cells.
To examine the effects of ATP depletion on cell substrate interactions, we explored both the adhesion of suspended cells and the release of adherent cells. In both cases, 90% ATP depletion resulted in a significant loss of adhesion that was totally reversible after stimulation with HGF, despite no affect of HGF on cellular ATP levels. In contrast, 99% ATP depletion resulted in a more severe loss of adhesion that was only modestly affected by treatment with HGF. In a similar approach of examining cyanide-mediated ATP depletion in MPT cells, Kroshian et al. (22) found that 94% ATP depletion for 1 h resulted in a loss of 66% of the previously adherent cells and that this could be reversed by allowing ATP levels to recover for 30 min. They demonstrate disruption of the actin cytoskeleton after ATP depletion and suggest that regulation of the actin cytoskeleton may be critical for the process of adhesion in this setting. Indeed, work by Raman and Atkinson (38) confirms the disruption of actin stress fibers with ATP depletion and has demonstrated that expression of constitutively active Rho can partially reverse this effect (38). Interestingly, full reversion of the ATP-depleted cell phenotype to normal did not occur with expression of constitutively active Rho because the dispersion of vinculin (which forms the site for anchoring stress fibers to focal adhesions) and paxillin from sites of focal adhesions was not reversed.
Examination of the morphology of mIMCD-3 cells revealed that whereas control cells had extended processes and spread out 2.5 h postplating, the majority of ATP-depleted cells remained rounded. In the setting of stimulation with HGF, this failure to extend processes and flatten was partially reversed in 90% ATP-depleted cells but only minimally altered after HGF treatment in severely ATP-depleted cells. This result, coupled with the HGF-mediated increase in adhesion to the integrin substrates fibronectin and type 1 collagen, suggested to us that HGF increased adhesion by regulating integrin-dependent cell spreading and that stimulation with HGF could overcome a loss of integrin-dependent cell spreading in the setting of moderate ATP depletion.
After the binding of HGF to c-met, multiple signaling pathways are activated, including the phosphatidylinositol 3-kinase, phospholipase C-γ, src, and growth factor receptor-bound protein 2 (grb2) (4,7). The association with grb2 mediates activation of the ERK1 and ERK2 members of the MAPK signaling pathway as well as a unique association with GAB1 and its membrane localization (27,51). We have demonstrated that activated ERK2 can bind and phosphorylate GAB1 at the membrane after HGF stimulation (41), and active ERK has been recently demonstrated to associate with focal adhesions at the sites of cell attachment after src activation (11). We therefore examined the possibility that ERK activation and subsequent localization to focal adhesions at the cell membrane may be an important mediator of HGF-induced cell adhesion.
To inhibit ERK activation, we utilized the selective MEK inhibitor U0126 (10). A dose-response curve in mIMCD-3 cells revealed that 20 μM U0126 caused complete inhibition of HGF-induced ERK1 and ERK2 activation (20) but had no effect on tyrosine phosphorylation of c-met or GAB1 or on activation of the PI 3-kinase after stimulation with HGF (Yu C, unpublished observations), and this dose was therefore chosen for the present experiments. In the setting of inhibition of ERK activation, HGF failed to cause an increase in adhesion in either control or ATP-depleted cells, strongly arguing that the increase in adhesion after stimulation with HGF requires activation of the MAPK-signaling pathway. In agreement with this, HGF failed to activate ERK 1 or 2 in 99% ATP-depleted cells, a setting where HGF also failed to reverse the loss of cell adhesion. The substrate for the MAPK pathway in HGF-induced cell adhesion has yet to be identified. Recently, stimulation with HGF in breast carcinoma cells and hepatocytes has been found to cause tyrosine phosphorylation of FAK with an increased linkage between integrin complexes and the actin cytoskeleton (1, 33). Whether FAK phosphorylation is regulated by ERK activation in mIMCD-3 cells is as yet unknown.
In summary, HGF and its receptor have been found to be upregulated in the kidney after ischemic renal injury, and the administration of HGF diminishes the degree of tubular epithelial cell loss and/or promotes tubular repair. By using an in vitro ATP depletion model to partially mimic the effect of ischemia, we demonstrate the loss of matrix attachment by otherwise viable epithelial cells, and make the novel observation that this effect is completely reversed by HGF in moderately ATP-depleted cells. The ability of HGF to reverse the ATP-depletion-induced cell detachment is dependent on activation of the ERK-signaling pathway. We therefore propose that at least part of the mechanism by which HGF decreases injury after renal ischemia involves the promotion of sustained adhesion of sublethally injured cells that would otherwise detach and undergo apoptosis.
This work was supported by National Institute of Diabetes and Digestive and Kidney Diseases Grant DK-52897 to L. G. Cantley.
Address for reprint requests and other correspondence: Z.-X. Liu, Section of Nephrology, Yale Univ. School of Medicine, 333 Cedar St., LMP 2093, New Haven, CT 06520 (E-mail:zhen-xiang.liu{at}yale.edu).
The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.
- Copyright © 2001 the American Physiological Society


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MyEssayWriting.com - Essay Writing Services.
Fairy Tales
by Wilhelm Hauff
(Rare Book Collection)
A collection of eleven fairy tales in English, written by Wilhelm Hauff, illustrated by Arthur A.
Dixon, and published by E. Nister circa 1910 in New York.
* The Story Of Caliph Stork
* Fatima's Rescue
* The Story Of Little Mouk
* The Story Of The False Prince
* The Dwarf Long-Nose
* Abner, The Jew Who Saw Nothing
* The Story Of Almansor
* The Story Of The Florin
* The Cold Heart
* The Adventures Of Said
* The Cavern Of Steenfoll: A Scottish Legend

subset: commoncrawl


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Who can wear the jewelry?
Yogis, fashionistas, surfers, meditators, nine-to-fivers, executives, professional hippies, moms, dads, aunts & uncles, teens... the list goes on. The jewelry can be worn by anyone seeking a calmer mind, body, and spirit.
How is the jewelry unique?
Each piece of jewelry is hand-made in Bali. One of things that makes our malas unique is that the beads are received without holes in them- fresh off the trees! They are drilled from the top, where the bead was attached to the tree, to the bottom. The bead is also threaded the same way to maintain the prana, or energy.
What is the significance of having 108 beads in my mala?
All of our malas are made to specific spiritual numbers. The number 108 is significant for numerous reasons. Some believe there are 108 stages on the journey of the human soul, while others believe there are 108 desires in mortals, matched by the same amount of lies and delusions. Some associate the possibility of enlightenment with taking only 108 breaths a day, while in deep meditation. Others connect the number's significance to the heart chakra, as there is said to be 108 energy lines leading to it, one of which is believed to be the path to self realization.
How do I use my mala?
Malas are often used to help count mantras during meditation. What's a mantra? It's a word or sound repeated to aid in your concentration during meditation. To count your mantras, hold your mala in your hand, and turn each bead with your thumb. Reaching the guru bead during meditation, the bead that dangles from the mala, signals a time for reflection. Do not continue over the guru bead. Instead, turn around and continue in the other direction.
Are these the actual malas from Eat, Pray, Love?
Yes, as a matter of fact they are! Julia Roberts donned the malas in the movie while filming in India and Ubud, Bali. She was also featured in O Magazine wearing one of our gold malas, which can be ordered upon request.
Where do rudraksha beads come from?
Our rudraksha jewelry is made from rudraksha beads. They are grown inside of what looks like a blueberry on trees in South East Asia. Our authentic beads are sustainably harvested and fairly traded.
What does rudraksha mean?
The name rudraksha stems from Hindu mythology and means "the eyes of Shiva." It is widely believed that the god meditated on the welfare of mankind and emerged crying tears of compassion. Upon hitting the earth, the tears crystallized to form the trees.
What are the healing qualities of rudraksha beads?
It's believed that there are many healing qualities associated with wearing rudraksha beads. Many people believe the beads help alleviate stress, depression and lethargy. Others have said the beads reduce heart disease, lower blood pressure and increase mental clarity. Ancient Hindu scriptures have proclaimed that the beads bless one with peace of mind, protect against evil doers and spirits, and grant nirvana, moksha, peace, and prosperity.
Are there different types of rudraksha beads?
The rudraksha bead varies in size from 3 mm to 40 mm. The smaller beads are the most rare. The beads have a very long life, sometimes known to last up to eight generations. Most importantly, each bead has a number of facets or serrations on the face, which are called mukhis and or mukhas. We mainly use five mukhis, which is the bead that represents lord Shiva, and is the most commonly sought after.
What are the healing qualities of Sandalwood beads?
In ancient Buddhist tradition, Sandalwood has been celebrated as the perfect meditation tool for thousands of years. It is widely believed that the sandalwood scent can stimulate sensuality, grant a sense of awareness, invoke tranquility, awaken the divine thoughts within and promote profound relaxation. Burning sandalwood incense can benefit healing work by promoting a deeper relaxed state in the healer, increasing spiritual energy.
How do I care for my rudraksha / sandalwood jewelry?
Rudraksha & Sandalwood beads are sacred and need to be treated with respect and care. We suggest you wash them every so often to remove dirt and dust, and to oil them with coconut or sandalwood oil for preservation. Due to your natural body oil, they will turn darker with wear, which is the desired effect. It's best to remove the jewelry when sleeping.
Will my rudraksha / sandalwood jewelry look the same as in the picture?
Each piece of jewerly is hand made, using hand-picked rudraksha or sandalwood beads and gemstones based on their vibrations and energies. Therefore, each gemstone is unique and may vary in colour and pattern. But rest assured, the rudkrasha or sandalwood beads and gemstones will still carry their believed qualities.
Will the rudraksha / sandalwood jewelry cure my problems?
While many believe in the healing qualities of rudraksha and sandalwood beads, as well as the benefits from certain gemstones, Mala Collective does not intend for these products to treat, cure or prevent any disease. The results are the sole responsibility of the wearer. Please consult your own physician regarding the treatment of any medical condition. This information is not a replacement for a thorough consultation and examination from a licensed health professional.
All prices are listed in USD and do not include applicable taxes or shipping fees. The risk of loss for all merchandise ordered on this website passes to you the buyer when the merchandise is delivered to our shipping carrier. Please contact us at email@example.com to request a tracking number, and we will gladly add it to your order. We are not responsible for lost or stolen packages and/or missing items that do not have a tracking number or are in process with the courier or in the case it has been delivered, does not have a signature at delivery point.


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A LAWYER has argued that a sex act between a convicted rapist and his 20-year-old female victim was consensual, because the woman didn't "close her legs".
George Pule, 34, was found guilty of raping the young woman in an alleyway after the bouncer promised to help her gain entry to a club.
The lawyer, Keith Jeffries, told a hearing at the Wellington District Court the woman's allegations were made out of regret and not because the sex was non-consensual, Fairfax NZ reports.
"All she would have had to do was to close her legs ... it's as simple as that," he said.
"Why didn't she do that? ... The reason she didn't do that was because the sex was consensual, as easy as that."
Mr Jeffries' comments were immediately reprimanded by Natalie Gousmett of the Wellington Rape Crisis group.
"It's disgusting, but also unnecessary, to use that kind of victim-blaming rhetoric."
"No, she didn't fight back, she didn't scream her head off, she didn't go running into the street screaming 'Rape!' But this isn't an American TV show.
"This is real life. She was scared, and she didn't want to make the situation worse."


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Price: $21.99
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Description: Help your young fan show off his love for the Orlando Predators with this Victory tee. This awesome tee feature bold Orlando Predators graphics for a look hes sure to love. Its the perfect thing to sport in the stands on game day.
Date Added: 8/21/2016 12:36:51 PM


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|Google Scholar output for 3bm7
The CC_2132 (NP_420935.1) gene from Caulobacter crescentus encodes a protein that belongs to the ABM (antibiotic monooxygenase) family (PF03992 ). The structure adopts a ferredoxin-like fold that belongs to the dimeric alpha+beta barrel superfamily. Most of the homologues from DALI search are hypothetical protein with unknown function. CC_2132 has been directly compared with 2OMO, 1LQ9 and 1N5Q. CC_2132 is a monomer in each asymmetric unit. Based on the interface calculation with/without the tag at the N-terminal, it should be a dimer as the biomolecule.
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text: just came back from new york, there was a no gi tournament held at columbia university where basically people from college bjj clubs from all over came to compete against each other. I did 155 lbs and under advanced and won the division (2 matches). My first opponent was a purple belt from columbia (renzo's) I beat him 2-0 takedown. And second opponent was the wrestling coach of columbia who i beat by advantage (this guy had more fury than anybody ive ever rolled with ). I have the matches on video, will post them soon. Best part about it was that it only cost 5 bucks to compete, and I won a 20$ gift certificate to budovideos.com Anybody got reccomendations on what I should buy? edit: videos up, and turns out i beat the second guy 2-0 as well, guy who was taping it just told me that i got points for a sweep at the end match 1: http://hezkezl.com/cage/ken1.wmv match 2: http://hezkezl.com/cage/ken2.wmv


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kudrna family
I am searching for a William Kudrna born about 1871-1873 in bohemia/czechoslavika, immigrated to the united states as a young boy with is father. They immigrated to Red Cloud, Nebraska. William Kudrna married a Josephine Havel in Nebraska and they had 4 children. Robert J. Kudrna, Ralph W., Elsie G.,and Wilma L. or I.. In the 1930 census this family was still located in Red Cloud, Nebraska. If anyone has any information on this family I would be greatly appreciative if you would contact me.I have hit a brick wall and don't know where else to look. Thank you in advance for any and all help.
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Re: kudrna family
alice burley 11/05/04
Re: kudrna family
Kris Iveans 11/07/04
- Re: kudrna family


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With smartphone designs growing bigger by the day, the Steelie HobKnob Kit gives you a better way to hold and display your phone using the ingenious Steelie magnetic mounting technology.
The kit contains two components: the Steelie Magnetic Phone Socket, and the HobKnob Handle. The Magnetic Socket adheres to the back of any phone (or case) with 3M® VHB™ foam adhesive tape. With the Magnetic Phone Socket on your device, the HobKnob handle can then magnetically attach in a snap to create an ergonomic handle or mini stand for your device. The Steelie HobKob Handle consists of a specially engineered aluminum knob topped with a grippy rubber ball that fits securely around the socket and makes a comfortable and secure handle. Perfect for one-handed viewing, showing videos without obscuring the screen or taking selfies without any fumbling in the process. The HobKnob makes a useful angled stand, in either landscape or portrait, for typing, viewing, reading, or video chatting.


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When you are looking for the best bagels and coffee in Calverton, Bean & Bagel Café is your one-stop eatery for breakfast and lunch items - at a reasonable price. Co-owned by John and Joanne Leibold, Bean & Bagel Café offers Suffolk County customers high quality, homemade food at a reasonable price. Bean & Bagel Café serves a wide variety of breakfast and lunch items: bagels, daily specials and so much more. Dedicated to providing personalized and friendly service to customers each and every day, Bean & Bagel Café is your home away from home. We also provide catering!
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The framework calls this member function if CWnd has the input focus and the WM_SYSKEYUP and WM_SYSKEYDOWN messages are translated.
It specifies the virtual key code of the Control-menu key. (For a list of of standard virtual key codes, see Winuser.h)
When the context code is 0, WM_SYSCHAR can pass the WM_SYSCHAR message to the TranslateAccelerator Windows function, which will handle it as though it were a normal key message instead of a system character-key. This allows accelerator keys to be used with the active window even if the active window does not have the input focus.
For IBM Enhanced 101- and 102-key keyboards, enhanced keys are the right ALT and the right CTRL keys on the main section of the keyboard; the INS, DEL, HOME, END, PAGE UP, PAGE DOWN, and arrow keys in the clusters to the left of the numeric keypad; and the slash (/) and ENTER keys in the numeric keypad. Some other keyboards may support the extended-key bit in nFlags.
This member function is called by the framework to allow your application to handle a Windows message. The parameters passed to your function reflect the parameters received by the framework when the message was received. If you call the base-class implementation of this function, that implementation will use the parameters originally passed with the message and not the parameters you supply to the function.


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HCM CITY — In a move to welcome the start of the summer season, Vietjet has announced the start of three golden days, in which one million promotional tickets will be on sale, priced from as low as zero dollars, at www.vietjetair.com.
The promotion, which is available on all domestic routes, continues Monday through Wednesday from noon to 2pm. Tickets may be used from May 1 to December 20.
Payment can be made with debit and credit cards, including Visa, MasterCard, JCB, KCP and American Express, as well as ATM cards issued by 29 banks in Viet Nam that have been registered for internet banking.
"To prepare for coming trips this summer, April is the best time for travellers to "hunt" for affordable tickets," the airline said.
With promotional tickets good throughout the year, Vietjet is expected to offer its passengers with a memorable and exciting travel season. — VNS


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How do I get there?
Directions to:
- CLOSED: Co Chi Vegan
- 1 Werbellinstrasse (at Karl-Marx Strasse), Berlin, Germany, 12053
- 015772039058
Note: Driving directions do not work in all areas of the world.
If this is not working for you, consider going directly to Google.
Please Report Incorrect Markers.


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The event was postponed in August because of Hurricane Irene. Metro said it was postponing its scheduled work on the tracks for Sunday, Oct. 16, the day of the event.
Metro had planned to work on the Red, Blue, Yellow and Orange lines. Part of the Red Line was also going to be closed between New York Avenue and Fort Totten stations.
"We will adjust our plans to accommodate the rescheduling of this one-in-a-lifetime event," said Richard Sarles, Metro's general manager, in a press release. "While the rebuilding efforts of the Metrorail system are critical, we recognize the national significance of this event and will reschedule our work for a later date to ensure that customers have a seamless travel experience."
Metro is spending $5 billion over the next six years on a massive capital improvement plan. Part of that involves repairing and rebuilding track work and implementing recommendations from the National Transportation Safety Board, following the 2009 fatal crash on Metro's Red Line.
On Oct. 16, Metro plans to open at 5 a.m, two hours earlier than normal, operate eight-car trains and offer free parking at all rail stations.
The transit authority said it plans to have "customer service ambassadors" on hand at stations to help guide people to the memorial.
Follow me on Twitter @postmetrogirl .


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About Me
Natalia Romano, a Los Angeles native, is a self-taught couturier and a rising young designer in the West Coast fashion scene. At age fourteen, she attended the summer fashion program at OTIS School of Art and Design in Los Angeles where she was one of the youngest students to enroll. While in high school, Natalia was accepted to the Fashion Institute of Design and Merchandising (FIDM). As the youngest student in her class again, Natalia stood out as a creative and energetic young talent. As a student in the prestigious advance study program, Natalia was asked by Mattel to design a gown in celebration for 50th anniversary of Barbie. Her winning design is a one-of-a-kind gown that
was reminiscent of Barbie's first 1950's bathing suit. Natalia additionally took on other projects, designing a dress for the unveiling of the Los Angeles Kings' new team jersey and Warner Brothers for the 70th anniversary of the Wizard of Oz. Later working with Disney and Colleen Atwood, for the release of Tim Burton's Alice in Wonderland.
Soon after graduating, Natalia Romano established her own company, debuting her ready-to-wear collection in Spring
2009. Natalia Romano collections are now available in boutiques worldwide.
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|Facts collected from tagged text on this page
|Facts about this page
It's tough to say which player did more for short people in basketball, Muggsy Bogues or Spud Webb. The former was the shortest professional athlete in the modern era in a sport that encourages height, but the latter, Anthony "Spud" Webb, may have done more when he won the 1986 NBA Slam Dunk contest. Standing just 5-foot-7, Webb flew through the air like a hummingbird to win the title.
Before his moment in the limelight, Webb had been a solid player at N.C. State. The Texas native went on to play nine seasons in the NBA, and according to videos on YouTube, was still dunking a basketball well into his 40s.


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Chapter 13 : The Party - Part One
|Rating: 15+
|Chapter Reviews: 15
Background: Font color:
-Chapter 13-
Lyrics – Undercover Martyn by Two Door Cinema Club (OMG yay!) as sung by Sirius Black ;D
Warning; Substance use - Alcohol
The Potter household…
James's P.O.V
"Yeah, up there. That banner that way. No, not that way! Peter! The hell are you doing man!?"
"I-I'm so sorry Prongs!"
"Gah! Just…go eat cheese or something will you!?"
"Yeah, o-okay!"
And with that, rat man scurried into the kitchen hastily and out of my sight. I knew I wasn't being fair on him, I was just so frustrated, and any of the other guys would most likely punch me if I ordered them about like that. Or something alike. And Moony with his werewolf strength hurts.
I blew my fringe up in frustration.
Sirius hopped down off the chair he was using to 'survey the room as we decorated' and slapped me on the back consolingly.
"Really man, is she worth this?" he asked gently.
I paused for a second and deliberated. Only for a second.
"Without a doubt. Padfoot, I'm just worried that she won't come. I have no idea what I'll do if she doesn't, and I know Lily. She most likely won't. And you guys will have your girls, which is great for you, but a massive reminder to me that the girl I'd die for hates my guts."
"Dude, she prolly doesn't hate your guts. I mean, they may look a little icky, but on the whole they do their job nicely!"
I shoved the grinning shaggy haired boy slightly and shook my head, a little smile playing around my lips. But my thoughts then inevitably returned back to the beautiful red haired love of my life. Who probably won't show up to my party I invited her to. Although, on the off chance she did, I even bought some chicken; her favourite party food.
I was also worried about appearances. If Lily didn't show up, then people would most likely realise something's up and start who knows what rumours. And I didn't need that. Plus, I'd look so sad to be the only bloke there, at my own party, without a date. Why must love be so cruel?
If I was honest, I still didn't completely understand what I had done wrong. Sure, maybe I shouldn't have said I saw a fit girl. Even if I was talking about her, she wouldn't have realised that and would've taken it the wrong way. But even if she had taken it that way, why had it affected her so much? There was one answer I could guess. But I didn't dare jump to that amazing conclusion. I was hesitant to even think it, just in case I'd get my hopes up and be wrong. But it was still there…
Lily liked me too.
There. I thought it. Could that amazing possibility be reality? I mean, she had said that kiss had meant something to her, right? She must like me a little. And if the reason why she's not speaking to me right now is because she thought I was looking at other girls, she must like me a lot…
Did I just think that? Bullocks! I can't take it back either! I'm gonna be jinxed now, she won't love me! Dear Merlin I'm falling apart. Look at what you do to me Lily…if only I had the guts to tell you it all…
"Oi Prongs, I dunno if you've realised, but I'm kinda still up here…" Remus called from inside my living room's chimney.
"Oh yeah, sorry. D'you think it's clean enough for the people using floo powder?"
"Yeah, totally! ….So can I come down now?"
"Sure Mooney, I guess so."
A few seconds later, a rather dusty looking Remus appeared, falling from my chimney coughing and spluttering.
"Ho ho ho!" Sirius greeted him back.
"Tell me I can get changed before doing anymore?" The sandy haired boy pleaded with me.
"Yeah, of course."
And with that, the dusty, unimpressed werewolf left the room to change. I glanced around my newly decorated living room, considering the décor in place. There were a couple of banners strewn along the walls, nothing cheesy, just red, yellow and blue. No green mind. The fancy leather squishy sofas had been pushed back against the walls to create a big dancing area, the high tech stereo currently where the TV had been. Little Japanese lanterns had been charmed to float near the ceiling, so gentle red would glow for lighting, and then they'd change colour. Purpley red and then to lime green…
"I think we should do the kitchen now Paddy." I said, distracting myself and smirking a little, knowing Sirius hated that name. And I got the desired reaction.
"James! It's Padfoot! Pad-foot! Get it!?" Said Padfoot howled and stomped his foot, following me to the kitchen.
Ahhh that had made me significantly happier. Nothing more amusing than taking digs at the drama queen.
"Right, the theme in this room is-"
"Alcohol?" Sirius asked hopefully.
"…no actually. Because we aren't having any alcohol, are we Sirius?" I said rather loudly, knowing my parents would hear that.
"That's right James!" Sirius added in the same loud tone. "And you are very irresponsible for suggesting it Peter!"
The 'culprit' in question looked up alarmed from the block of cheese he was eating.
I and Sirius stared at him, cheese bits covering his mouth.
"Dude, I wasn't being serious…"
Lily's P.O.V
I winced again as Tracy tugged the brush through my hair once more. I wasn't actually sure how she was styling it, I just knew that it better look bloody stunning, or I would be chasing the hair tugger in question with the brush all night.
Alright, so I wasn't in the best of moods. So shoot me.
My nerves were thrumming with something that felt like…nervousness. Excitement. A little hint of want…
No hormones, behave. We don't like that stupid hazel eyed marauder anymore, remember? That's right. We do not want that gorgeous, tempting, devilishly handsome, amazing kisser…
Not one bit. Nice to know I have everything under control.
Oh who am I kidding? I was a wreck! Underneath I was anyway. Truth was, I was wanting that damn marauder more than ever. And I didn't know why. Why was he still not talking to me? That wasn't right. And the longer he left it, the more I wanted to talk to him…to see those beautiful eyes cast over me once again, just like that night…
Oh sod off! Since when were you aloud to rule my body anyway hormones? I'm sure my brain never agreed to this, 'cause if it did, it's clear I didn't actually have one. Gah, listen to me! I'm having an argument with myself. You stupid worthless marauder! You're screwing with my brain!
"Lily, are you alright…? You look like you want to punch someone…" Beth's concerned voice shot through my internal argument, and I realised how much of a lunatic I must have sounded. Even to myself.
"Yeah, I'm alright thanks Beth."
"Are you still worrying about tonight? Merlin Lily, I thought we were over that."
"Tracy, I'm not over it, I still reckon I'll be the awkward turtle of the group. I'm not looking forward to it."
She scoffed.
"That's not what you're worried about. You're worried about bumping into him aren't you?"
Alright, so what if I was? That wasn't even the scary thing. No, the scary thing was that a part of me wanted to bump into him…I was worriedly unsure of how big that part may be… And I resented it. Therefore resenting that gorgeous marauder who was making me feel this way!
Can you say, conflicted? Little bit.
"Well, I know you want to look good in front of him, whether you're worried about seeing him or not." Tracy smiled at me in the mirror, filling in my silence.
I glanced gratefully back at her. You have to love your friends, right?
"Yeah I do. Though it's gonna be some work on your part." I teased.
"Be quiet! Have you seen yourself? You're bloody stunning! And I'm gonna help that prat notice it, or my name isn't Tracy Stone. Believe me babe, when he sees you, he's not going to be looking at any other girl, fit or not."
"Yeah! Oh, I can do your make up for you! It says here in this magazine that guys this month like the 'vanilla look'. Sort of soft looking. I'll do that bit for you if you'd like! And add a bit of my vanilla spray over here for added smelling effect!" Beth beamed at me, happy to help as always.
"Guys, you are just awesome." I told them.
Tracy nodded in agreement and grinned.
"Right, Beth, you get ready in your dress now while I finish Lily's hair. When that's done, I'll put my dress on and you can do Lily's make up. Then when that's done, me and you can do our make up and take your rags out, while this one here puts her dress on. Deal?" She planned, organising us.
"Deal." I and Beth nodded enthusiastically while the latter hopped off her bed and went to get ready into her dress.
Oh Merlin. And the nerves strike back with double the vengeance…
Later on in the common room…
"Don't be ridiculous woman, come on out!" Tracy hollered at me from the other side of the common room door.
"No! I look stupid! Who bought this dress? What was she thinking? I look like a green cream puff!"
"The ruffles aren't that bad Lily and you bought the dress so you must've liked it." Beth reasoned.
"The sales woman was really convincing okay? I didn't try it on beforehand, and now I look like a tool."
"You won't do at all. Now get your butt out of that room now, or we'll be late and I will not be impressed!" Tracy demanded, and she was right. We were going to be late.
I sighed.
"Fine! Just…give me a minute…"
Tracy huffed unappreciatively. I swallowed a gust of breath to calm me down, then twisted the handle and exited the room. Tracy and Beth beamed at me as I appeared.
"You're rocking that dress and half girl!"
"You're just being nice." I told them while trying to pull down the hem of my dress.
It was quite short, only stopping mid-thigh, leaving the rest of my legs feeling bare. Lime green – my favourite colour – was the dress; it was tight-ish and ruffled slightly at the top, black lace outlining the dress. I felt weird in it. Oh why hadn't I tried it on first? Damn it. At least Tracy had done my hair beautifully (looks like there was to be no chasing with a brush) curled at the ends and twisted into the most complicated bun, slightly to the left side of my head, so little stands could fall onto my shoulder. And my shoes were gorgeous in my opinion, the same lime green and quite tall in the heel; a little bow on the back of them with tiny diamonds in the middle of them. I still felt a little silly in my get up.
Tracy meanwhile looked stunning in hers, bluey-black like her nails; her dress flowed out at the back down to her calves while the front stopped at her mid-thigh too. Strapless and with no pattern it was simple yet stunning and striking. Typical Tracy Stone. Her shoes tied up with ribbons that wound round her legs to her knees and the heel was taller than mine. Her brunette hair was simply straightened, to add to the simple stunning effect.
Beth was stood at the side of her, clad in a pretty lemon yellow dress. She'd gone for a pastel soft look to match her personality, the yellow folds complimenting her body shape well, stopping just above her knees. A single strap slung over one of her shoulders, leaving the other bare, which looked pretty on her, showing her delicate skin. The ringlet rags had worked as her hair curled in tight little ringlets, spilling over her shoulders and down her back. Surprisingly, Beth's shoes were the highest of all of ours (which she later explained that the only nice looking yellow shoes she could find were these ones, so she'd accepted them, not caring if she broke her neck just so she could look good in front of Remus.) And they were open toed pretty kitten heels.
The sandy haired Marauder would go crazy for her. In fact, Sirius too would probably resemble a dog when he saw Tracy with his tongue lolling out. But me…? I didn't know. I wasn't sure if I liked what I looked like, never mind the hazel eyed boy I was attempting to impress…
No, I wasn't trying to impress him! God damn it. I could see this night being a long and difficult one…
"Right, let's take this party by storm!"
And with Tracy's declaration, we headed off to the Potter Mansion… Gulp…
James's P.O.V
The party…
"Man, you know how to throw a party!" My shaggy haired best friend whooped as he and his quickly increasing conga line bounced past me. I laughed and high-fived him, determined not to look like the worrying misery guts I was on the inside.
Damn it.
Lily and her friends still hadn't arrived yet, and the party was in full swing; people partying in every space downstairs I could see, the loud music hyping everyone up, helping them have a good time. Although Remus and Sirius didn't appear to look in the slightest bit worried that their dates hadn't yet arrived, I was kinda freaking out about it. At least I had an excuse; I knew my girl had a chance of not showing at all. And I didn't know how I'd react to it if she didn't.
Huh, 'my girl'. I smirked a little to myself imagining Lily's reaction if she knew I'd called her my girl. Probably a string of obscenities, followed by an "In your dreams Potter." Ahhh, she was so lovely. And funny. And smart, and utterly wonderful…
Christ I needed to get a hold of myself.
But where was this wonderful girl now? I could barely wait to see her. And besides, I was running out of excuses to not accept the offers of a dance from the other girls at my party. And I think I may be seriously close to getting punched by the dates of the some of the girls who asked me.
Well hey, it isn't my fault I'm incredibly sexy, is it?
But if only Lily would see me that way…
"Hey, get this down you." Remus suddenly appeared at my side with a red plastic cup in his hand, filled with a dark liquid. I smiled and nodded at him.
"Cheers mate. But fire-whiskey…?" I took the offer and glanced curiously at my friend. Remus wasn't usually one to encourage drinking, especially the fire-whiskey sort. And also especially after Sirius once had drunk so much that he had mistaken Remus for a very pretty girl…
Sirius tended to lose what little sense he had when drinking fire whiskey. That's why we do our best to hide it from him.
"Yeah, I figured you needed something to calm those nerves." He smiled at me, also leaning against my living room wall as I was.
Avoiding his gaze, I continued to observe the party in front of me, watching people laugh, dance, drink and flirt to high heaven.
"I have no idea what you're talking about." I attempted to achieve one of my signature Potter smirks at a random dancing girl, and she smiled and fluttered her lashes back at me.
Sheesh she was hideous. Although, I may just think that because she's not the fiery haired girl I was seeking.
"Right, of course you don't. And I thought you said brunettes weren't your type?" He asked, nodding at the girl I'd 'flirted' with, an 'I know aaaaall about you' look plastered on his face.
"That was last year."
"Of course it was."
Tch, honestly! When he wasn't being a paranoid, grizzly were-wolf, he was an annoying 'know it all' all seeing guy. Right now I wasn't sure what I'd prefer more.
"She'll be here soon Prongs." He patted my shoulder.
As nice as he was being, I really didn't want to talk about Lily for once. It'd just make me more depressed, at my own party too. I didn't want that, I wanted to get away. Then suddenly a new song came on and inspiration struck me.
"What was that Moony? I can't hear you!" I yelled, cupping my ear in imitation of not being able to hear him properly, despite the fact I was stood next to him.
Confused yet willing to console, he attempted to repeat his sentence a tad louder.
"I said, she'll be here-"
"What!?" I cut him off.
"She will be-"
"It's no use! I'll talk to you later Moony!"
And with that, I left a very annoyed looking Remus behind, leaning against the wall. I thought my acting had been brilliant. However, Moony didn't seem to have appreciated it. I don't think he had believed me. His loss, I thought I was a true actor in the making there.
Dodging the dancing people, I made my way to the kitchen, seeking solstice in there. However I was only met by more of my dancing peers, who were excessively drinking as well. I sighed and poured myself some more fire-whiskey from behind the sink (where we'd hid it from Padfoot) Ah well, if you can't beat 'em, join 'em.
Lily's P.O.V
We could hear the music as we arrived outside the house, walking up to the front door. There were people outside the house already, laughing and/or snogging. I wrinkled my nose as we walked past a particularly wild couple.
"Cheers Tony!" Tracy waved goodbye and thanked our lift as he nodded and went to meet his friends. Tracy had gotten him to apparate us here, through means of persuasion. Ew, not like that you dirty minded people!
"You'll be fine Lils." Beth squeezed my hand as she realised I was breathing a little heavier than usual. Grateful, I squeezed her hand back as thanks. Man I was kinda dreading this.
"Party time!" Tracy hollered, and swung the posh double doors back to reveal a busy scene of dancing, laughing, chatting people already, and we were only in the hallway.
"Where d'you reckon they'd be?" Beth asked aloud as we stepped over the threshold.
Kitchen." Me and Tracy answered in unison, and we set off, me a little lagging behind. I wasn't so eager to find the hazel eyed marauder. We weaved in and out of the rowdy people in the mansion, getting wolf whistled and cat called as we passed. Tracy loved the attention, waving here and there. Beth just went red, bless her heart. She didn't like it. I couldn't care less on the other hand. Though, they were mostly drunk anyway, which slightly irked me a little. They were annoying. Finally, we made it to the kitchen after the long trek of getting there.
Seriously, could his house be any bigger?
Gulping, I quickly surveyed the scene… but to my disappointment, no Potter was to be found…
Wait! No! Not disappointment!
Oh dear Lord.
There was however, a suspicious looking Sirius standing near the sink. We made our way over to him.
"Black! What are you up to?"
He jumped about a mile in the air as he heard Tracy's voice, then sheepishly answered;
"Don't scare me like that Stone! You'll give me a right…heart…attack…"
As predicted, Sirius turned into dog mode as his eyes met Tracy's figure. It was seriously like watching a cartoon, eyes popping out, jaw hitting floor, tongue rolling out a mile… you know the drill.
"Yes Black?" She smugly observed her finger nails, smirking.
He suddenly regained his 'cool' and stood up properly.
"Well, you certainly scrub up nice, don't you Stone?"
"Hmm, glad you think so. You're not so bad yourself… if you tucked that bit of your shirt in."
Sirius seemed to have matched his messy hair with his look; a nice blue and white checked shirt, half tucked, half un-tucked, with his trendy blue jeans. He looked messy, yet cute at the same time. And he knew it.
"Nah, it's the fashion Stone! And hey you girlies, you're looking pretty fine yourselves girlfriends!"
"Um, thanks Sirius. You look pretty…uh, fine, tonight too." Beth attempted to compliment the strange marauder.
"Cheers Sirius. You look awesome." I answered simply, smiling.
He beamed at me, pleased.
"Um, Sirius?" Beth began. "Do you maybe know where I can find Re-"
"Padfoot! If you're in here, I'm gonna-" We never found out what Remus was gonna do to Sirius, because as he charged into the kitchen looking like thunder, he caught sight of Beth and stopped in his tracks.
And dither girl strikes back!
"Leave it!" I muttered under my breath as Beth went for her fringe, her face warming a few million degrees.
"Beth! Wow! You look…amazing!" Remus came over to our group, holding his arms out for Beth to delicately fall into, which she did do. They shared a happy hug as Sirius and Tracy looked on like proud parents smiling, and I stood there. Like a plank.
I knew I'd be the dang awkward turtle! However, in the moment of hilariousness, I had to prevent myself from doing the awkward turtle with my hands, just to cheer myself up. I'd only look more of a weird loner if I was wiggling my thumbs to myself.
"So, shall we go to the living room then?" Remus stared hard at Sirius, who hiccupped and looked highly embarrassed.
"I'm gonna have to find James to hide that bottle again. Just stick to the bloody butter beer Sirius, you can handle that. I seriously don't want a replay of last time."
"Right, to the living room!" Sirius cried over Remus, and we headed to the designated area, wondering what did happen 'last time'.
As we made our way over to the living room, I couldn't help but check everywhere I could for any sign of the messy haired marauder who hadn't yet appeared. I thought I was inconspicuous, but it seems I'd been busted as Beth made a point of asking for James when we bagged ourselves a corner of the room.
"You guys seen James then?" Beth asked, nudging me slightly.
Gee. Thanks. Point attention to the needy loner everybody! Yoo-hoo! That's me!
"Yeah, he's been about. Though, I'm not sure where he is at the minute, he kinda ditched me." Remus frowned and sighed as he recalled.
"Yeah, he seemed a little weird. He didn't want to join my conga line!" Sirius admitted sadly.
Tracy humoured him and patted his back consolingly, as Remus's frown deepened slightly.
"Just when did you find that bottle of whiskey…?"
"Um…uh…well…'AND SHEEEE SPOKE, WORDS THAT WOULD MELT IN YOUR HAND'!" Sirius suddenly started bellowing out the lyrics to the current track playing, and he wasn't in tune either.
"Sirius! Hey! Oi! Siri- ugh. Forget it." Remus face palmed as the shaggy haired marauder continued to sing his heart out. Badly.
Beth giggled a little at the scenario, and Remus smiled at once as he heard the sound, turning his face towards her so he could see her pretty face laughing. They were lost in their own world, smiling and giggling to each other. I knew what would come next, those laughing lips meeting…
I couldn't watch. But then to my horror, as I turned to face Tracy, I found that she'd already gotten one step ahead of Beth and Remus, and was kissing Sirius like there was no tomorrow. Even if it had been a desperate bid to shut him up, I was stood on my own.
Argh! Awkward turtle/plank isn't enough to cover what I felt like! Never mind a third wheel, how about a lone wheel when the rest of the wheels in the whole world were being used? Am I not good enough for the bike? Has the bike forgotten about this wheel…? Damn it Lily, enough bike metaphors, get out of there!
"Um, I need to…yeah." I coughed awkwardly, and began to leave.
"Oh, Lily! You don't need us to come with-"
"No! Uh, no, I'll be fine, I'll um, catch up later."
And with that I spun round and left the happy couples before I puked. Seriously, could they get anymore gross, 'in your face Lily'?
I sighed as I left the living room, knowing they weren't doing it to torture me. They were just having fun and liking their relationships. It was hardly their fault I had thrown a fleggy at James. Ugh, why did I have to be such a hot headed fool? We could have just talked about it at least…
Lost in my thoughts, I bumped into someone, some lanky kid with greasy blonde hair who grinned at me.
"Hello there lady, wanna dance?"
"Uh…no." I told grease ball and continued down the impossibly long corridor, thankful he wasn't following me.
"I know you want me!" He called after me.
Ugh seriously!? Now people were looking at me! Amazing. Brilliant. Thank you grease ball, thank you. I just hoped he was someone's friend from Drumstrang or something so no rumours started. Although if I was honest, he didn't look DS material.
Grumbling to myself I walked along down the corridor, trying to focus on something else other than my humiliation. And that's when I realised I'd walked straight down snogging street. Seriously, the walls were littered with lip locked couples.
Oh nice one. Thank you Merlin! You sure know how to make me feel wanted! Stupid karma! Fine, the next time I see that bloody marauder, I'll snog his face clean off, is that what you want?
Brilliant. Now I'm talking to myself some more. Just have another internal argument why don't you Lily? Ugh! You're losing it! Go somewhere!
Angry tears threatened to overspill from my eyes and I suddenly broke into a run, finding some stairs and deciding to leg it up them, away from the kissing couples. Ignoring the 'off limits' sign, I reached the top of the stairs and was welcomed by a much needed quiet. Only a faint thrumming of the music from way downstairs could be heard.
The hallway upstairs was huge as well; I couldn't see the end of the left or the right side of the hall, although the lights weren't on so that probably would have helped. I could still see quite well though, despite the darkness, and I took a moment to observe my beautiful surroundings. The celling just above the stairs where I was stood, curved outward into a dome shape, beautiful swirls chipped into it. Royal red wallpaper covered the hall walls; a golden dido rail separated the wall in half, looking very majestic in gold. A red plush carpet surfaced the floor that I was suddenly aware of, as I realised I was most likely digging holes into it with my heels.
"Oh bugger!" I mumbled and kicked them off quick, deciding to carry them instead. My bare feet were now in contact with the carpet, and it felt like it had never been stood on in its entire existence. I wriggled my toes in the fabric for a minute, enjoying the soft feel on my already hurting feet. I then looked up from the carpet and my eyes spotted something.
A golden frame hung on one of the walls, the fancy swirls outlining the picture it held. The picture itself was of a small boy who was hard to make out, so curious, I stepped closer. He came into focus, his little face beaming, his tiny hands outstretched happily with five year old joy, clutching a little toy train in one of them. Dressed in oversized robes, his messy black hair on end, his hazel eyes shining…
I gasped, lifting my finger and placing it on the little boy's hand.
His dad was kneeling beside him, looking happy and proud at the camera, laughing at his five year old boy wearing his old Hogwarts robes…
Unknowingly, a smile tugged at my lips as I stared at this little boy's face, his joy radiating through the picture to me. So innocent, so young, so happy… A nice James Potter…
Standing up, I shook myself a little and stepped away from the photograph. It'd had a weird effect on me, and I was keen to go sit somewhere and sulk to myself. But I wasn't sure which way to go, I'd never been upstairs before. Deliberating, I decided to just drop my shoe on the floor and let that decide.
With a soft thud, the shoe landed and the toe pointed to the right. That's where I set off. After picking my shoe up of course, you numpties.
"Hmmm, three." I decided after seeing the hallway stretch right down, doors popping up left right and centre. I closed my eyes, spun round in a circle once or twice with my hand outstretched, then stopped. Opening my eyes, I saw I had picked the right side of the hall way again. I counted three doors down, and wearily put my hand on the golden polished doorknob. Ah what the hell, may as well.
I opened the door and walked in, flicking a light switch and shutting the door behind me. The surroundings now illuminated, I found myself in a bedroom. I wasn't sure if it was a spare bedroom yet, but it looked a little too personalised to be one…so I decided to snoop, praying this wasn't Potter's parents love nest or something.
Now that would be an experience I'd love to forget.
The room was also red and gold. I could certainly see this family were proud Gryffindor-ians. A large king sized bed stood proud in the centre of the room, red bed covers neatly pulled over it, plenty of golden pillows at the head of it and a gold throw placed over the foot of the bed. Red gossamer curtains were either side of the bed, ready to be drawn shut when the person wished to sleep. Perhaps this was a guest room for Gryffindors? There may be others in the house decorated for Hufflepuffs and such. Regardless, I continued to snoop, heading for the dark polished wooden draws at the other end of the large room.
I opened the top draw to find neatly folded shirts. Perhaps this room had been taken by a guest already. But who? Deciding to shut the draws as they'd most likely be full of clothes also, I headed for the bed to search under it. Sticking my head under, I put my hand out in front of me and felt something metal. Quickly I withdrew my hand with the item in question clutched in it. As I slowly unfurled my hand, a worn blue colour became prominent. I cupped the item in my hand, and saw that it was a blue train, the paint worn off in some places it was that old.
"Aw." I smiled, turning one of the wheels with my finger, when I was suddenly reminded of that picture outside…
"Holy crap!" I screeched, dropping the train suddenly.
James's room. I was in James's room!
Well of course. Out of all the millions of freaking doors down two possible hallways, of course I'd end up here.
I'll be honest; I was pretty pissed off at karma right now.
No, it's okay Lily! Just leave. He'll never know you were in here. Quickly I rose to my feet, not bothering to put the train back under the bed and leaving it on top of the bed instead, and headed for the door.
Which opened as I reached it. Revealing James of course, his hazel eyes blinking in wonder.
Aw nuts!
A/N; Hello thar! What do you think? I have honestly been typing this for ages. I have had an essay a week, and essay a week! How unfair? But I carried on typing thanks to you awesome readers/reviewers out there! I had to stop the chappie here, I had originally planned to continue, but then I saw the word count... but on the plus side, the next one should be written pretty quickly!
Anyway, little explanation over! Please leave a review if you've read, always fills me up with the happies when I read them! And encourages writing! ^^
Happy New Year everyone! =D
Love, Alie xxxxxxxx
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A Game of Truth or Dare: The Party - Part One
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Revolutionizing the Functionality of APPS for the Benefit of Merchants and Consumers Worldwide.
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New regulation seeks to ban 50 top invasive species in Europe
A harmonised approach to tackling Europe's 50 most damaging invasive species has been proposed. NGOs would like more species included
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or call 020 8267 8120


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Vehicle PricingEst. Payments $244/mo*
Selling Price: $16,995
Popular Features
Contact Seller
This Vehicle Is Located At
J & R Auto Sales
2800 W. 12th Street
Sioux Falls, SD
Estimate Payments
|Extended Warranties
Financing Available
|Fog Lights
|Wheels - Aluminum
|Rear Defrost
|Trailer Hitch - Receiver
|Front Bucket Seats
|Center Console
|Overhead Console
|Rear Bench Seat
|AM/FM Radio
|CD Player
|CD Changer
|Premium Audio System
|Air Conditioning
|Cruise Control
|Tilt Steering Wheel
|Intermittent Wipers
|Power Windows
|Power Locks
|Power Mirrors
|Power Driver Seat
|Power Passenger Seat
|Remote Keyless Entry
|Heated Mirrors
|Heated Seats
|Satellite Radio
|Side Airbags
|Roof Airbags
|Power Steering
|Airbag - Driver
|Airbag - Passenger
|Traction Control
|Rear Backup Sensors
|Oil Pressure Gauge
|Outside Temp Display
|Trip Odometer
|Volt Meter
|Water Temp Gauge
|Body Color Side Molding
|Tinted Glass
|Tow Hooks
|12V Power Outlet
|Center Storage Armrest
|Door Pocket/Bins
|Drivers Foot Rest
|Floor Mats
|Fold Down Rear Seat
|Front & Rear Cupholders
|Illuminated Entry
|Low Oil Level Warn Light
|Passenger Assist Handles
|Rear Bench Seat
|Rear Seat Center Armrest
|Remote Fuel Filler Door
|Steer Wheel Audio Cntrls
|Steer Wheel Cruise Cntrl
|Vanity Mirrors
*Default estimated payment is based on 3% rate for 60 months and $3,399 down payment. Estimated payments are for informational purposes only and don't account for acquisition fees, destination charges, tax, title, and other fees and incentives or represent a financing offer or guarantee of credit from the seller.


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Prices, promotions, styles, and availability may vary. Our local stores do not honor online pricing. Prices and availability of products and services are subject to change without notice. Errors will be corrected where discovered, and Lowe's reserves the right to revoke any stated offer and to correct any errors, inaccuracies or omissions including after an order has been submitted.
Minwax 8-fl oz Wood Conditioner
Item # 506352 Model # 134074444
Product Information
- Specifically designed to use before staining to help prevent streaking and blotching and help ensure a beautiful, even stain
- Use on any wood but is especially necessary when working with soft or porous woods like pine, alder, birch, and maple
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04-03-2008 #1Administrator
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PSN authentication on PlayStation Forums coming April 14
The official European PlayStation Fourms have revealed that from Monday 14 April onwards, you will need to log in to the Forums with your PSN Sign-In ID (email address) and PSN password.
In preparation for the change, the Eu PlayStation Forums will be offline for one week from midnight on Sunday 6 April until noon on Monday 14 April.
To quote Lestrade (Community Team Coordinator): I have an update for you on the introduction of PLAYSTATION Network authentication on the PlayStation Forums: we will now be launching this new functionality on Monday 14 April.
From Monday 14 April onwards, you will need to log in to the Forums with your PSN Sign-In ID (email address) and PSN password. Your Forum name (i.e. the name that will appear on your public profile and next to your forum posts) will be your PSN Online ID.
If you haven't already, please make sure that you have linked your Forum account with your PSN. Guidance on how to do this can be found here. But don't worry if you don't manage to link your accounts in time, you will be able to log in to the forums and post using any valid PSN account.
To allow us to update the Forums in preparation for this change, they will be offline for one week from midnight on Sunday 6 April until noon on Monday 14 April.
Once again, we're very sorry that this roll-out was delayed from its original launch date but we'll look forward to seeing you back in the Forums after Monday 14st April.
The Sign-in ID used to authenticate and log in to your PLAYSTATION®Network account will be the email address you register for PSN. This is different to your PSN Online ID, which is the equivalent of your Forum Username.
Your Sign-in ID will not be displayed in the Forums, on your Forum Profile or be otherwise accessible to any other person.
To protect your account and to keep your PSN account secure, we strongly recommend that you do not disclose your PSN Sign In ID (your email address) to anyone on the Forums - whether in a public thread or via Private Messaging. And *NEVER* tell anyone your password, even if they claim to be a Sony employee or representative. Doing so could compromise the safety of your PSN Account.
More PlayStation 3 News...


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World Media
Qatar closes its bombed health center in Syria
The Qatar Red Crescent says four bombs were dropped from a helicopter on the hospital in the rebel-held neighborhood of Sakhour on Saturday. The state new agency SANA said 20 were killed and 55 were wounded in the wedding. A suicide bomber killed at least 22 people Monday in an attack targeting a party in the northeastern province of Hasakeh, the Syrian Observatory for Human Rights and medics said.
World Media
Poll shows Clinton taking lead in Florida
In the new results, Clinton was at 45 percent, Trump 41 percent, Libertarian Party candidate Gary Johnson 5 percent, and Green Party nominee Jill Stein was at 2 percent. Forty-two percent of independent likely voters said Clinton bested Trump in the debate, while 27 percent gave Trump the edge. The numbers are from Quinnipiac, which released a slate of state-level polls.
World Media
Kylie Jenner, 19, Buys Fourth California Mansion at $12M
The Cape Cod style mansion, also has a gourmet kitchen, the flawless location for domestic goddess Kylie to experiment with her latest dishes, a swimming pool and a home cinema. "I'm going to post the meme he just sent me right now". Over the course of the very long Q&A, the Kylie Lip Kid creator also touched upon her new hair color, explaining that it had actually been in the works for a while, which is why she hadn't dyed her hair in a year.
World Media
Date announced for Manchester's Olympic and Paralympic parade
Rio de Janeiro, September 19: The closing ceremony of Rio Paralympic Games 2016 turned into a feast of light, colour and music in the Maracana stadium with the theme "Biodiversity: Our Inspiration". Of the Canadian athletes who medalled, four were under age 20. International Paralympic Committee President Sir Philip Craven, addressing the event said Rio Paralympics were " uniquely Brazilian and wondrous ", as quoted by the BBC.
World Media
Yogi Adityanath backs Salman, says India's fight is against terrorism, not art
On Arnab Goswami's NewsHour Debate, Mahesh Bhatt was asked about his views on the current ban on Pakistani actors in India. "Can they not come up with either a decision or a comment on what Thackeray is saying?" A few days ago, superstar Salman Khan had invited the MNS wrath after he had said, "Pakistani artistes are not terrorists".
World Media
Jewish Leader Dumps Labour Party After Jeremy Corbyn Is Reelected
Jeremy Corbyn was re-elected head of the British Labour Party, leading to the resignation of a Jewish official from the party. "That would be the gross betrayal of the Labour party and our people". She said Labour must try to keep "the best of what we have in the European Union, not just because it's good for Britain, but because it's good for Europe and it's good for the world as well".


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Wyoming Technology Business Center
1938 Harney
Laramie, WY 82072
Phone: 307-766-6395
Email: firstname.lastname@example.com
e2e Wyoming is an educational networking program with chapters in Laramie, Sheridan, and Casper designed to improve the climate for the start-up and growth of entrepreneurial companies. Five meetings are held each year where entrepreneurs meet with other entrepreneurs and business service providers. Ideas are shared, plans are developed, and many partnerships are formed.
Thursday, September 14, 2017
5:30-7:00 pm
The Lyric (230 W Yellowstone Hwy)
Travis Todd, co-founder of FullContact, will be the next e2e Wyoming speaker in Casper on September 14th. FullContact is a Denver-based technology company that provides a suite of cloud-based contact management solutions for businesses, developers, and individuals. Launched from a basement in 2010 by Bart Lorang, Dan Lynn and Travis Todd, FullContact has graduated from TechStars accelerator in Boulder, received $50M in venture capital and now employs 230 people.
Travis will tell the story of launching FullContact and the company's growth over the years, which includes acquiring four companies along with the assets of two others. FullContact now has a presence in Denver, Dallas, Latvia and India, giving it a truly global presence. FullContact, which introduced the idea of "paid, paid vacation" in 2012 has consistently been voted as one of the best companies to work for by Outside Magazine.
In addition to serving on FullContact's board of directors, Travis's current role is Head of Labs, which allows him to work on experimental, research and development projects. Travis has a BS in Mechanical Engineering from the University of Wyoming and an MBA from the University of Texas at Austin. He and his wife Mindy have two children Houston (6) and Elsie (2), with a third child on the way. Raised on a family ranch in northeastern Wyoming, Travis and his family were excited for the opportunity to recently return to this great state.
Wyoming Technology Business Center
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Phone: 307-766-6395
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Go to Encyclopedia of Life...
Nash, T.H., Ryan, B.D., Gries, C., Bungartz, F., (eds.) 2002. Lichen Flora of the Greater Sonoran Desert Region. Vol 1.
Squamules: up to 4 mm wide, rounded to elongate, adnate or ascending near the margin, rarely imbricate, weakly concave to plane upper surface: medium brown to dark brown, dull, epruinose or faintly pruinose, smooth to sparingly fissured margin: concolorous with upper side, straight to weakly up-turned, entire or lobed upper cortex: up to 200 µm thick, composed of rather thin-walled hyphae with more or less round lumina, not containing crystals of lichen substances, sometimes containing calcium oxalate (mainly in the epinecral layer) medulla: not containing lichen substances or calcium oxalate lower cortex: of mainly periclinally oriented hyphae, not containing calcium oxalate lower surface: pale to medium brown, K- Apothecia: up to 1.5 mm diam., laminal, convex and immarginate even when young, (medium-) dark brown to black, epruinose ascospores: ellipsoid, 9-14 x 5-7 µm Pycnidia: laminal, immersed conidia: bacilliform, 6-9 x c. 1 µm Spot tests: upper cortex and medulla K-, C-, KC-, P- Secondary metabolites: none detected. Substrate and ecology: on soil and rock, mainly in deserts, up to 1830 m World distribution: western North America Sonoran distribution: locally common in Arizona, California, Baja California and Baja California Sur. Notes: The species is sometimes difficult to distinguish from P. globifera and P. tuckermanii. It differs from the former in its often smaller and more closely adnate squamules with a more smooth, dull, and paler brown upper surface, and in having more thin-walled hyphae with more rounded lumina in the upper cortex. The stainable layer of the upper cortex is roughly 10-15 cells thick in P. globifera and 15-25 cells thick in P. luridella. The apothecia of P. luridella are dark brown to pure black, rarely medium brown, and always epruinose; those of P. globifera are medium to dark brown, rarely black, and sometimes yellow pruinose. Psora tuckermanii differs morphologically in having generally larger, usually somewhat paler and more pruinose squamules and medium brown, sometimes white or yellow pruinose apothecia. Furthermore, it usually contains calcium oxalate in the medulla. Timdal ([1986] p. 274) regarded some specimens from California and Baja California with dark brown to black apothecia and lacking calcium oxalate in the medulla as belonging to P. tuckermanii; these specimens are here regarded as belonging in P. luridella.


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Golden Retriever
12 reviews from the community
Review of "Golden Retriever"
"Never ever buy a blue carpet !!!"
In my defence the carpet came before the dog and although i have to hoover on a daily basis sometimes i get to go 2 days without having to hoover!
I have 3 cats who were here first and a 9 year old boy all of whom have adapted to Reggie very well and in turn he has to them! He does chase them but as a game he has never hurt them he just wags his tail and off they go down the garden at full speed it is the funniest thing to watch!
He is such a loving dog he always looks as if he is smiling and he loves to play.
We got him from a breeder and he is registered as a pedigree but unfortunately he does have hip displacia which retrievers are apparently prone to, we did tell the breeder and she was most upset as it was the first case she allegedly had had, but as she wasn't breeding again from the same dogs it won't be a problem in the future i hope for more litters. It doesn't seem to affect him now but maybe in his later years it will. I look at it that we were given him as a gift because if she had known he would have been put down (she told me this) when he was born. It means he is limited to the amount of daily walking and exercise but have you ever tried to stop a retriever from running and jumping around?! especially one that in his head he is still a puppy?!
The good factors of Reggie is he is loving, happy, great with the kids and the new baby, I was worried he may be a little jealous but he has just accepted her as if she was always there (he keeps trying to give her his squeaky toy!), she gives him the biggest smile!
They have very soft mouths, apparently its why they're used as gun dogs as they won't damage the 'catch' we experimented and even though he was never trained to do it we gave him a raw egg to carry and he did without so much as a crack he didn't even drop it! He is one of the family not just a dog!
The disadvantages of Reggie is mainly the hair and although i am told they are very trainable ours seems to have fallen thru that into a big loop hole! He will give left paw and right paw (for treats usually), he will even do 'high fives' he will sit, he won't lay down unless he wants his belly rubbed, and if you walk him he pulls like mad, he is better off the lead but then when he sees someone he wants to play with or say hello to he will just run off he won't come back until he is ready not even for food we have tried! And if you have ever wanted the model back garden like on the tv programmes forget it ours looks more like baghdad at the moment he loves to dig! You also have to watch the wagging tail which rarely stops and always knocks everything flying! his whole bum wags with the tail.
But everyone loves reggie he gets so much fuss and love from everyone who lives near us and when he plays his houdini trick and escapes thru yet another escape hatch he has dug in the garden people just laugh and bring him home again tail wagging big grin on his face!
I am sorry you just can't be angry for long they are beautiful, friendly and loyal family dogs and i wouldn't change him for the world!!
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karenes published 01/10/2004He sounds lovely! I'd love to get a dog again but it'd be locked up all day which isn't fair. A lot of pedigrees have problems like that. My mum has a west highland terrier who has terrible eczema. I wince everytime I see him! K x
MAFARRIMOND published 08/09/2004A beautiful dog. Maureen
Minha published 08/09/2004I NEARLY had a 'goldie' a few years back - but finished up with border collies! (AND six cats!). If he does have trouble with his hip - try a Bioflow collar. Hazel xx
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Listed on Ciao since: 18/06/2002