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|Table of Contents
|Vol. 50, No. 3, 2013
Issue release date: July 2013
|Section title: Research Paper
Systemic Administration of Thrombin Peptide TP508 Enhances VEGF-Stimulated Angiogenesis and Attenuates Effects of Chronic HypoxiaOlszewska-Pazdrak B.a, b · Carney D.H.a, b
aDepartment of Biochemistry and Molecular Biology, The University of Texas Medical Branch, and bChrysalis BioTherapeutics Inc., Galveston, Tex., USA Corresponding Author
Prof. Darrell H. Carney
Department of Biochemistry and Molecular Biology
The University of Texas Medical Branch
301 University Blvd., Galveston, TX 77555-0645 (USA)
E-Mail firstname.lastname@example.com
Revascularization of chronic wounds and ischemic tissue is attenuated by endothelial dysfunction and the inability of angiogenic factors to stimulate angiogenesis. We recently showed that TP508, a nonproteolytic thrombin peptide, increases perfusion and NO-dependent vasodilation in hearts with chronic ischemia and stimulates NO production by endothelial cells. In this study, we investigated systemic in vivo effects of TP508 on VEGF-stimulated angiogenesis in vitro using aortic explants in normoxic and hypoxic conditions. Mice were injected with saline or TP508 and 24 h later aortas were removed and cultured to quantify endothelial sprouting. TP508 injection increased endothelial sprouting and potentiated the in vitro response to VEGF. Exposure of control explants to hypoxia inhibited basal and VEGF-stimulated endothelial cell sprouting. This effect of hypoxia was significantly prevented by TP508 injection. Thus, TP508 systemic administration increases responsiveness of aortic endothelial cells to VEGF and diminishes the effect of chronic hypoxia on endothelial cell sprouting. Studies using human endothelial cells in culture suggest that protective effects of TP508 during hypoxia may involve stimulation of endothelial cell NO production. These data suggest potential clinical benefit of using a combination of systemic TP508 and local VEGF as a therapy for revascularization of ischemic tissue.
© 2013 S. Karger AG, Basel
Therapeutic angiogenesis Vascular endothelial growth factor Thrombin peptide Rusalatide acetate Endothelial dysfunction Hypoxia Endothelial cells
TP508 is a 23 amino acid synthetic peptide representing residues 508-530 of human prothrombin which was identified as a potential receptor-binding domain based on competition for high-affinity thrombin binding to fibroblasts [1]. In contrast to thrombin, TP508 has no enzymatic activity and does not promote or interfere with blood coagulation [1]. Studies have shown that TP508 accelerates healing of dermal wounds [2,3,4], fractures [5] and bone defects [6,7]. TP508 is being evaluated in human clinical trials as a therapeutic drug for treatment of diabetic foot ulcers [8] and for acceleration of fracture repair [9]. TP508 acceleration of healing is associated with stimulation of early revascularization in dermal and bone injury models [2,3,5,10]. From these studies, however, it is not clear whether TP508 stimulated angiogenesis by itself or promoted angiogenic responses to other factors present in the injured tissue.
VEGF is an important angiogenesis factor that is produced by hypoxic cells during tissue injury to stimulate and direct angiogenesis [11,12]. A number of studies have shown that VEGF-stimulated endothelial angiogenic responses in vitro [13,14,15,16,17] and angiogenesis in vivo [18,19] require activation of endothelial nitric oxide synthase (eNOS). Although VEGF administration increases revascularization of ischemic tissue in experimental animal models, only limited benefit was achieved with VEGF therapeutic angiogenesis in clinical trials in patients with myocardial ischemia [20,21,22]. Thus, this limited effect of VEGF in patients with coronary artery disease suggests decreased endothelial cell responsiveness to VEGF in chronically hypoxic tissues. We have shown that exposure of human endothelial cells to chronic hypoxia attenuates VEGF-stimulated signaling and angiogenic responses [23]. We recently demonstrated that TP508 activates eNOS and stimulates production of NO in human endothelial cells [23]. Using a pig model of chronic myocardial ischemia, we demonstrated that TP508 increased myocardial perfusion and function, and restored NO-dependent vasodilatation and NO production in arterioles isolated from ischemic myocardial regions [24]. More recently, it has been shown that TP508 decreased infarct size, improved NO-dependent microvascular function and increased activation of eNOS following acute myocardial ischemia-reperfusion injury in normal [25], hypercholesterolemic [26] and diabetic pigs [27]. These studies led us to hypothesize that TP508 protects endothelial cells from hypoxia-induced loss of endothelial function, and potentiates responsiveness of vascular endothelial cells to VEGF.
The purpose of this study was to examine systemic effects of TP508 on VEGF-stimulated angiogenesis in vitro using aortic explants and to determine whether TP508 administrated in vivo protects endothelial cells against effects of chronic hypoxia. Our results demonstrate that TP508 intravenous injection increases endothelial cell sprouting and enhances VEGF-stimulated sprouting from aortic explants isolated from mice 24 h after injection. Moreover, chronic hypoxia markedly inhibits basal and VEGF-induced endothelial cell sprouting, but TP508 injection protects endothelial cells from this hypoxic effect. These results suggest potential therapeutic benefit of a combination of VEGF and TP508 for myocardial revascularization or treatment of peripheral artery disease.
Thrombin peptide TP508 (AGYKPDEGKRGDACEGDSGGPFV, also known as rusalatide acetate CAS No. 87455-82-6) was synthesized and purified by American Peptide Company (Sunnyvale, Calif., USA). Human recombinant VEGFA165 was purchased from R&D systems (Minneapolis, Minn., USA). Matrigel Matrix (phenol red free) was obtained from (BD Biosciences, Bedford, Mass., USA). L-arginine, NaNO2, and NaI were purchased from Sigma (St Louis, Mo., USA).
All experiments were approved by the UTMB Institutional Animal Care and Use Committee. Twelve- to 15-week-old ICR male mice (Charles River Laboratories, Wilmington Mass., USA) were housed (5 per cage) in a controlled environment with a 12-hour light/dark cycle, with temperature maintained at 21 ± 0.5°C and a room humidity of 50 ± 20%. Water and chow were provided ad libitum. Mice were placed in a restrainer and their tail veins were injected with a single 500-µg dose of TP508 in 100 µl of saline or with 100 µl of saline alone (placebo control). A single injection of TP508 (500 μg/mouse) was chosen for these studies based on dose response studies of TP508 injection on ex vivo aortic endothelial cell sprouting and in vivo biological effects where injection of 500 µg/mouse showed maximal effects [manuscript in preparation]. After 24 h mice were euthanized by CO2 inhalation and aortas were removed as previously described [28,29].
Angiogenic endothelial cell sprouting was studied by culturing mouse aortic explants on Matrigel Matrix (BD Biosciences) as described previously [28,29]. Twenty-four hours postinjection, thoracic aortas were isolated from TP508- or saline-administrated mice and transferred to culture dishes containing cold endothelial basal medium (EBM; Lonza, Walkersville, Md., USA). The periaortic fibroadipose tissue was removed under a dissecting microscope and aortas were rinsed with cold EBM and cut transversely to create 1 mm aortic rings (∼10 per aorta). Aortic rings were cut, opened and the inner endothelial surface was placed directly on Matrigel Matrix coated wells of 24-well plates. Aortic explants were cultured in endothelial growth medium which contains EBM supplemented with 5% fetal bovine serum and SingleQuots (Lonza) in 5% CO2 at 37°C. After 24 h of culture, aortic explants from control or TP508-injected mice were stimulated with VEGF (50 ng/ml) or vehicle and cultured in normoxia. To determine the effects of hypoxia, aortic explants from control and TP508-injected mice were incubated with or without VEGF under hypoxic conditions (1% O2, 5% CO2) for 2 days and then switched back to normoxia. Endothelial cell sprouting was monitored daily by inverted phase-contrast microscopy (Nikon, Tokyo, Japan) and images were captured using a Spot RT camera with Spot Advance image capture software (Diagnostic Instruments Inc., Sterling Heights, Mich., USA) at ×40 and ×100 magnification.
Image analysis and quantification for the area occupied by sprouting endothelial cells and maximal endothelial cell migration from aortic explants edges was performed using MetaMorph software (Molecular Devices, Downingtown, Pa., USA). For each experiment, we isolated aortas from three controls and from three TP508-injected mice. Aortic sprouting was quantified from triplicate culture wells containing a total of 6 aortic explants per experimental condition. The area of endothelial sprouting was normalized to the perimeter of aortic explants visualized in the field of observation. The area of sprouting from control mouse explants cultured in normoxia for 5 days was expressed as a value of 1.0. For maximum endothelial cell migration we measured the longest distance of migrated cells from the edges of aortic explants in 3 different regions from each explant.
Human coronary artery endothelial (HCAE) cells from Lonza were cultured in endothelial growth medium (EBM supplemented with 5% FBS and SingleQuots; Lonza) in 5% CO2 at 37°C. Two-day postconfluent HCAE cells, between passages 4 and 6, were cultured in normoxic or hypoxic (1% O2, 5% CO2 at 37°C) conditions and assayed for effects of hypoxia on VEGF expression, viability and NO production.
Quantitative SYBR Green real-time PCR was used to determine the expression of human VEGF mRNA. HCAE cells were cultured on 6-well plates. Two-day postconfluent cells were treated with TP508 (50 μg/ml) or vehicle and cultured in normoxic or hypoxic conditions for 24 h. Total RNA was isolated using RNAqueous kits (Ambion, Austin, Tex., USA). Isolated mRNA samples were coded for blinded analysis. Total RNA was reverse transcribed using Taqman Reverse Transcription Reagents Kits (ABI). Quantitative PCR amplifications were done using 2 μl of cDNA in a total volume of 25 μl using SYBR Green PCR Master Mix (ABI) on an ABI Prism 7000 Sequence Detection System. Primers for human VEGFA were sense: TAC CTC CAC CAT GCC AAG TG and antisense: GCG CTG ATA GAC ATC CAT GAA C. These primers recognize common sequence for all splice variant transcripts of human VEGFA. Reverse-transcribed cDNA cycle threshold values were calculated from triplicate sample determinations and normalized to 18S RNA.
Confluent HCAE cells were cultured on 24-well plates under normoxic or hypoxic conditions for 24 h. Normoxic and hypoxic cells were then stimulated with TP508 (50 μg/ml) or VEGF (50 ng/ml) or vehicle (control) in the same conditions. After 24 h of stimulation, the medium was replaced with EBM containing 200 μML-arginine (250 μl/well) and all cells were assayed for NO production during 1 h normoxic incubation. Supernatants were collected and analyzed for NO concentrations using a chemiluminescence NO analyzer (Model 270B, Sievers Instruments, Boulder, Colo., USA) as previously described [30]. The analyzer was calibrated for each experiment using a sodium nitrite standard curve. NO concentration in the EBM containing L-arginine was used as a background value and was subtracted from sample values to determine the amount of NO released from cells.
Analysis of variance (ANOVA) was used to determine if there was a significant difference among groups. A pair-wise comparison of groups was then done using the Tukey test. p < 0.05 was regarded as significant.
Stimulation of angiogenesis leading to tissue revascularization is required for normal wound healing after injury or surgical procedures [31]. The aortic sprouting assay is a well-established model for studying the effects of different factors on angiogenesis in vitro [28,29]. We used this model to determine the systemic effects of TP508 administered in vivo on angiogenic endothelial cell functions. Aortic explants were isolated from mice 24 h after TP508 or saline injection and cultured on Matrigel Matrix. As shown in figure 1a, endothelial cell sprouting was observed in aortic explants from control and TP508-injected mice after 4 days in culture. Explants from TP508-injected mice showed greater sprouting than controls at day 4 (fig. 1a). These sprouting endothelial cells grew progressively to form a branching network by day 5 characteristic of endothelial cells cultured on Matrigel (fig. 1a). Morphometric analysis of the area occupied by endothelial cells and maximal endothelial cell migration was significantly greater in aortic explants isolated from TP508-injected mice than in control explants on days 4, 5 and 6 (fig. 1b). For example, TP508-injected mouse aortic explants had a 6.4-fold increased area and 3.1-fold increased maximal migration of sprouting compared to explants from control mice at day 5 (fig. 3a). Thus, a single TP508 intravenous injection increased endothelial sprouting from aortic explants measured several days later.
VEGF is a key angiogenesis factor produced during tissue injury to stimulate and direct angiogenesis [11,12]. Thus, we examined the effect of in vivo systemic TP508 administration on angiogenesis stimulated by VEGF in vitro using the aortic explants. As expected, VEGF treatment of aortic explants from control mice resulted in increased area and migration of sprouting at all examined days compared to control, untreated explants (fig. 1a, b). We observed an 8.2- and 3.3-fold increased area and migration of sprouting, respectively, after 5 days culture in aortic explants from control mice in response to VEGF (fig. 3a, normoxia). This VEGF-stimulated sprouting from control explants at day 5 was comparable to the sprouting from aortic explants isolated from TP508-injected mice without VEGF stimulation (fig. 3a, normoxia). Importantly, VEGF stimulation of aortic explants isolated from TP508-injected mice showed significantly greater endothelial sprouting compared to aortic sprouting in response to VEGF from control mice on days 4, 5 and 6 (fig. 1a, b). These results demonstrate that in addition to promoting endothelial sprouting, TP508 injection also increases responsiveness of endothelial cells from aortic explants to VEGF-stimulated angiogenesis in vitro.
We have shown that exposure of human coronary endothelial cells to chronic hypoxia (1% O2) attenuates endothelial cell angiogenic responses [23]. The aortic angiogenic sprouting assay allows us to evaluate ex vivo effects of hypoxia on angiogenic responses of endothelial cells from intact vessel walls. This model also allows us to determine whether TP508 delivered in vivo can protect endothelial cells against effects of hypoxia.
Aortic explants isolated from control and TP508-injected mice were cultured for 24 h in normoxia to allow the explants to attach to the Matrigel and then switched to hypoxia (1% O2). In preliminary experiments we observed that continued culture of aortic explants under hypoxia completely inhibited endothelial sprouting, but if explants were cultured under hypoxia (1% O2) for 2 days and then returned to normoxia, measurable sprouting was observed. Explant exposure to hypoxia for 2 days markedly decreased the area of sprouting compared to the same explants cultured continuously in normoxia (fig. 2a, b, 3a, b). Moreover, VEGF-stimulated sprouting was suppressed during hypoxic exposure (fig. 2, 3a). At day 5 of culture, the area and maximal migration of endothelial sprouting in VEGF-stimulated control explants exposed to hypoxia was reduced 27- and 2.5-fold, respectively, compared to VEGF-stimulated sprouting in normoxia (fig. 3a). Hypoxia also significantly decreased endothelial sprouting in aortic explants isolated from TP508-injected mice compared to normoxic culture (2.3- and 1.6-fold; fig. 3a, b), however, the area and cell migration of sprouting at day 5 of culture was similar to sprouting from explants isolated from control mice cultured in normoxia (fig. 3a). In contrast to normoxia, there was no significant difference in area of sprouting from aortic explants isolated from TP508-injected mice in response to VEGF stimulation when they were exposed to hypoxia (fig. 3a). However, we observed a significant increase in maximal migration of endothelial cells in response to VEGF stimulation from hypoxic explants isolated from TP508-treated mice compared to explants from control mice (at day 4 and 5 culture; fig. 3a). These results show that exposure of aortic explants to chronic hypoxia markedly attenuates basal and VEGF-stimulated angiogenesis. Moreover, explants from TP508-injected mice that were exposed to hypoxia had levels of sprouting similar to that seen in control mouse explants cultured in normoxia. Therefore, TP508 injection protected endothelial cells from hypoxic effects.
To gain a better understanding of how TP508 may exert its effects on endothelial cells in vivo, we examined effects of TP508 on HCAE cells in vitro. We first determined TP508 effects on VEGF expression since increased VEGF production might explain the in vivo effects of TP508. Real-time PCR experiments showed that under both normoxic and hypoxic (1% O2) conditions, TP508 did not cause significant changes in VEGF mRNA expression levels during 24 h culture. Exposure of cells to hypoxia for 24 h increased VEGF mRNA expression 6.8 ± 1.3- and 7.5 ± 1.8-fold in control and TP508-treated cells, respectively (fig. 4b). These results showed that TP508 stimulation did not significantly affect the VEGF mRNA levels in normoxic or hypoxic cells. The concentrations of VEGF protein secreted into supernatants were also not significantly different between control and TP508-treated cells (data not shown). These results suggest that the in vivo effects of TP508 on endothelial sprouting are not likely to be due to TP508 changes in endothelial cell VEGF expression.
To determine whether hypoxia-induced inhibition of endothelial sprouting from aortic explants was due to endothelial cell death, we exposed HCAE cells to hypoxia (1% O2) for 48 h. Hypoxic cells remained confluent and cell viability was similar to cells cultured in normoxia (fig 4a).
Production of NO by endothelial cells plays an important role in VEGF-stimulated angiogenesis in vivo [18] and in vitro [32]. Therefore, we measured the amount of NO produced by HCAE cells during 1 h incubation, 24 h after stimulation with TP508 or VEGF. As shown in figure 4c, TP508 and VEGF stimulated 115 ± 33 and 111 ± 16 nM of NO production, respectively, compared to 41 ± 5 nM of NO released from control cells (fig. 4c). Exposure of cells to hypoxia for 48 h resulted in attenuation of NO production from control (25 ± 5 nM) and VEGF-stimulated cells (40 ± 17 nM). However, TP508-treatment of hypoxic cells increased NO production approximately 2-fold relative to control normoxic cells (79 ± 13 vs. 41 ± 5 nM; fig. 4c). Thus, hypoxia significantly inhibited VEGF-stimulated NO production, but had no significant effect on NO production induced by TP508. TP508-induced increase in NO production in hypoxic endothelial cells correlates with TP508-stimulated endothelial sprouting from aortic explants exposed to chronic hypoxia. This suggests that in vivo TP508 may have a similar effect on NO production which serves to protect endothelial cells from hypoxia.
The present studies were undertaken to determine the in vivo effect of thrombin peptide, TP508, on endothelial cell angiogenic responses under normoxic and hypoxic conditions. To address these questions we injected TP508 intravenously into mice and evaluated the effects of the peptide on angiogenesis using an aortic explant endothelial sprouting assay that mimics several stages of angiogenesis, including endothelial cell sprouting, proliferation, migration and tube-like structure formation [29]. Our results demonstrate that TP508 injection into mice 24 h prior to removal of aortas promotes endothelial sprouting and potentiates VEGF-stimulated endothelial sprouting. Exposure of aortic explants to chronic hypoxia markedly inhibited endothelial sprouting even after VEGF stimulation. However, aortic explants from TP508-injected mice exposed to hypoxia showed increased sprouting similar that observed in aortic explants isolated from control mice cultured in normoxia.
Effective wound repair is dependent on the number of cellular processes including revascularization of injured tissues [31]. TP508 has been previously shown to accelerate tissue revascularization and repair following tissue injury [2,3,5,7,10]. In a pig model of chronic myocardial ischemia, TP508 injection into the ischemic myocardium increased perfusion and restored myocardial function [24]. This treatment also increased NO-dependent vasodilation, expression of eNOS and NO production in arterioles isolated from ischemic heart regions [24]. We have shown that at the molecular level, TP508 stimulates rapid NO production in primary human endothelial cells [30] and prevents downregulation of eNOS expression in cells exposed to chronic hypoxia [24]. Intravenous injection of TP508 has recently been shown to reduce the infarct area following acute myocardial ischemia-reperfusion injury in normal [25], hypercholesterolemic [26] and diabetic pigs [27]. This systemic effect of TP508 caused us to question whether systemic administration of TP508 could enhance angiogenesis or protect endothelial cells from ischemia.
The aortic sprouting assay is a well-established model for studying the effects of different factors on angiogenesis in vitro [28,29]. We used this model in a novel manner to determine in vivo effects of systemically injected TP508 on vascular endothelial cell responses to ex vivo angiogenic factors and hypoxia. To determine whether a single dose of TP508 injected into mice promotes angiogenesis, we isolated aortas from placebo control or TP508-injected mice 24 h after injection and compared endothelial sprouting from aortic explants cultured on Matrigel Matrix. Our data show that TP508 injection significantly increased the area of endothelial sprouting and maximal migration of endothelial cells compared to aortic sprouting from control mouse explants.
VEGF is an endothelial-specific growth factor that is secreted by hypoxic cells at the site of injury to stimulate angiogenesis [11,12]. We therefore tested whether TP508 injection increases the responsiveness of endothelial cells to VEGF-stimulated angiogenesis. We found that VEGF stimulation of aortic explants isolated from TP508-injected mice showed significantly greater sprouting compared to VEGF-stimulated sprouting from control explants. These results demonstrate that TP508 injection not only promotes endothelial sprouting from aortic explants, but also potentiates responsiveness of aortic endothelial cells to VEGF.
Revascularization of chronic wounds and ischemic tissue is attenuated by endothelial dysfunction. Endothelial dysfunction induced by chronic hypoxia impairs angiogenesis in response to angiogenic factors [23]. In experimental animals, revascularization and recovery of blood flow to ischemic tissue is increased by administration of VEGF, but human clinical trials of therapeutic angiogenesis with VEGF were less successful than anticipated [20,21,33]. The differences between results in animals and clinical outcomes are postulated to involve different responsiveness of endothelial cells in young healthy animals used in laboratory studies compared to the endothelial cell responses in patients with chronic myocardial ischemia. Indeed, patients with coronary artery disease have endothelial dysfunction characterized by decreased expression of eNOS in endothelial cells [34,35] which contributes to attenuation of NO production and defective NO-dependent vasodilation in atherosclerotic vessels [36]. In the present study, we also evaluated the effect of chronic hypoxia on angiogenesis using aortic explants. Our preliminary results showed that continued culture of aortic explants under hypoxia (1% O2) completely inhibited endothelial sprouting, thus we exposed the aortic explants to hypoxia for 2 days followed by culture in normoxia. Exposure of aortic explants to hypoxia for 2 days inhibited endothelial cell sprouting and also markedly diminished VEGF-stimulated aortic sprouting compared to normoxia. These results are consistent with our previous report showing that chronic hypoxia decreases VEGF-stimulated signaling and angiogenic responses of endothelial cells [23]. Interestingly, hypoxia had much less effect on endothelial sprouting from aortic explants isolated from TP508-injected mice. In fact, explants from TP508-injected mice that were exposed to hypoxia showed sprouting that was equivalent to that seen in normoxic explants from control mice. These results suggest that TP508 systemic administration protects endothelium from the effects of chronic hypoxia.
To determine potential molecular mechanisms of TP508 on endothelial cells that may explain how TP508 systemic administration potentiates VEGF-stimulated endothelial sprouting ex vivo, we performed in vitro studies using primary HCAE cells. A first question was whether the effects of TP508 might be related to TP508-induced production of VEGF. We found that in HCAE cells TP508 did not increase VEGF mRNA expression or VEGF protein production (data not shown). These results are consistent with a previous study showing that direct TP508 treatment of microvessels in culture increased sprout elongation, but did not increase VEGF mRNA expression [37]. Thus, it appears unlikely that in vivo effects of TP508 are mediated through stimulation of VEGF production.
We previously demonstrated that TP508 stimulates NO production in human endothelial cells by different mechanisms than VEGF [30]. Since NO production is required for VEGF-stimulated endothelial cells proliferation [15], migration [13,32] and tube formation [38], it seemed possible that TP508-stimulated NO production may contribute to enhancement of VEGF-induced endothelial sprouting from aortic explants. Our in vitro studies showed that TP508 and VEGF stimulated NO production in normoxic HCAE cells to nearly the same extent. In hypoxic cells, however, basal levels of NO production and VEGF-stimulated NO production were significantly decreased while TP508 still stimulated NO production by 2- and 3-fold over that of normoxic or hypoxic controls, respectively. Although it is difficult to compare effects observed in cells in culture with events that happen in vivo, these results suggest that production of NO induced by systemically injected TP508 may contribute to the protective effects on hypoxia-induced inhibition of endothelial sprouting from aortic explants. Moreover, previous studies demonstrated that wound healing [39] and angiogenesis is significantly impaired in eNOS-deficient mice and is not improved by administration of VEGF [40], suggesting that NO production by endothelial cells is essential for angiogenesis. If TP508 injected systemically is enhancing angiogenic responses to VEGF and protecting endothelial cells from hypoxia, one could ask whether this effect is specific for VEGF or whether a TP508 injection might also enhance responses to other angiogenic growth factors.
Preliminary experiments demonstrate that TP508 systemic administration also enhances endothelial sprouting stimulated by FGF (data not shown). That the systemic TP508 effect on endothelial sprouting is not limited to a single growth factor pathway is consistent with the broad range of effects of TP508 that could be mediated by activation of NO signaling in endothelial cells to promote revascularization.
Previous in vitro studies have shown that TP508 stimulates elongation of sprouts from cultured microvessel fragments [37] and that TP508 increases attachment of endothelial cells or chemotaxis [41,42]. In these in vitro studies, TP508 was added to the medium, where it could have a direct immediate effect on the cells [37], or was attached to the plates or inserts where it could interact with integrins expressed on endothelial cells through an RGD domain in the peptide to stimulate cell attachment and haptotactic migration [41]. However, in our current in vivo studies, TP508 was injected into animals 24 h before aortic fragments were isolated and placed in culture. Thus, the effects of TP508 occurred during the period in which TP508 was in contact with the cells in the animal and persisted through isolation and culture without additional TP508 treatment. Thus, our studies indicate that TP508 induces changes in the endothelial cells that persist after isolation to provide longer-term protection from hypoxia and increased responses to angiogenic factors.
These results are consistent with the hypothesis that TP508 represents a natural breakdown product of thrombin that is released from fibrin clots into circulation to stimulate tissue revascularization and repair [43,44]. Studies are currently underway in our laboratory to identify the molecular mechanisms by which TP508 potentiates VEGF-stimulated intracellular signaling and endothelial cell functions in HCAE cells.
In summary, these results demonstrate for the first time that a single TP508 intravenous injection promotes angiogenic sprouting and potentiates VEGF-stimulated endothelial sprouting from aortic explants cultured under normoxic conditions. Exposure of aortic explants to hypoxia leads to inhibition of basal and VEGF-stimulated endothelial sprouting. Importantly, TP508 injection protects endothelial cells from the hypoxia-induced attenuation of endothelial sprouting. These results suggest a potential benefit of using a combination of systemic TP508 and local VEGF for treatment of chronic wounds and as a therapy for revascularization of ischemic tissues.
The authors thank Deborah Prusak and Dr. Thomas Wood of the UTMB Molecular Genomics Core Facility for real-time PCR analysis. The authors acknowledge research funding from The Texas Norman Hackerman Advanced Research Program and University of Texas Medical Branch Research Endowment Funds to D.H.C. Research reported in this manuscript was also supported in part by the National Institute of Allergy and Infectious Diseases of the National Institutes of Health (R43AI086135) and the National Cancer Institute (NCI/HHSN261201100035). Manuscript content is solely the responsibility of the authors and does not necessarily represent official views of the National Institutes of Health. Chrysalis BioTherapeutics Inc. of Galveston, Tex., and the University of Texas System own worldwide rights for commercialization of thrombin peptide TP508 (rusalatide acetate).
Prof. Darrell H. Carney
Department of Biochemistry and Molecular Biology
The University of Texas Medical Branch
301 University Blvd., Galveston, TX 77555-0645 (USA)
E-Mail email@example.com
Received: October 25, 2012
Accepted: January 12, 2013
Published online: April 16, 2013
Issue release date: July 2013
Number of Print Pages: 11
Number of Figures: 4
Number of Tables: 0
ISSN: 1018-1172 (Print)
eISSN: 1423-0135 (Online)
For additional information: http://www.karger.com/JVR
Copyright: All rights reserved. No part of this publication may be translated into other languages, reproduced or utilized in any form or by any means, electronic or mechanical, including photocopying, recording, microcopying, or by any information storage and retrieval system, without permission in writing from the publisher or, in the case of photocopying, direct payment of a specified fee to the Copyright Clearance Center.
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Orations on Philosophy and Education
By (author) Philip Melanchthon
Edited by Sachiko Kusukawa Christine F. Salazar
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Melanchthon by Philip Melanchthon
Book DescriptionPhilip Melanchthon (1497-1560), humanist and colleague of Martin Luther, is best known for his educational reforms, for which he earned the title Praeceptor Germaniae (the Teacher of Germany). His most influential form of philosophical writing was the academic oration, and this volume, first published in 1999, presents a large and wide-ranging selection of his orations and textbook prefaces translated into English. They set out his views on the distinction between faith and reason, the role of philosophy in education, moral philosophy, natural philosophy, astronomy and astrology, and the importance of philosophy to a true Christian, as well as his views on Classical philosophical authorities such as Plato and Aristotle and on contemporaries such as Erasmus and Luther. Powerfully influential in their time, inspiring many Protestant students to study philosophy, mathematics and natural philosophy, they illuminate the relationship between Renaissance and Reformation thought.
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Book DetailsISBN: 9780521583503
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Pages: 314
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Publish Date: 22-Apr-1999
Country of Publication: United Kingdom
Books By Author Philip Melanchthon
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This volume, first published in 1999, presents a translated and wide-ranging selection of Melanchthon's influential academic orations.
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subset: commoncrawl


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The leadership group approved recommendations of an SBC/BWA Study Committee revised in light of reaction to an earlier report released to the press Dec. 19, which chairman Morris Chapman said is now being described as an "interim" report.
In addition to calling for the SBC to withdraw from membership in the BWA and discontinue funding the global alliance, the "official report" also includes pledges to continue the SBC's funding commitment to the BWA through Sept. 30--the end of the 2004 fiscal year--and to invite select members of the BWA to meet with the study committee in Nashville prior to May 1.
Chapman, who is also president and CEO of the Executive Committee, said the meeting is in response to a letter from BWA president Billy Kim, a pastor in South Korea, and regional vice presidents of the BWA requesting a meeting to discuss reconciliation.
Since the SBC's growing discontent with the BWA dates back a number of years, Chapman said he doubted anything would happen at the meeting that would change the study committee's mind. Should the BWA delegation "repent," however, as one member put it in discussion, the Executive Committee would still have the option of reversing its vote at a meeting scheduled just prior to the start of the SBC annual meeting in June.
The study committee said negative reactions from other BWA member bodies since the December report "were actually a blessing, in that they served to demonstrate to all interested evangelicals why we had been experiencing increasing discomfort in attempting to define the SBC to the world through the BWA."
Some responses mourned the anticipated loss of Southern Baptists, the report said. But others, including the BWA general secretary and other leaders, "took the opportunity to vent what appear to be pent-up feelings of hostility about our convention."
"Due to these revelations, we need not now justify or vilify, but can simply do what we preferred to do in the first place, which is to politely withdraw from an organization that, at least for us, no longer efficiently communicates to the unsaved a crystal-clear gospel message that our Lord Jesus Christ is solely sufficient for salvation."
The report also noted that much has been said about a BWA vote to include the Cooperative Baptist Fellowship as a member body. "One soaked by rain need not blame the last raindrop," the committee said. "We strongly affirm the right of the BWA to determine its own membership and affiliations. It is that very right we now recommend that our convention exercise. The decision of the BWA to include the CBF merely served as a confirmation that we must, as a convention, allow the world to see us without having to look through a BWA lens--a lens which, for us, has become too cloudy."
The recommendation to withdraw membership from the Baptist World Alliance also calls for continued study on "how the Southern Baptist Convention may establish an even closer bond of fellowship with conservative evangelical Christians around the world."
It deletes the SBC $300,000-a-year contribution to the BWA effective Oct. 1, while reallocating the funds "to develop and execute a new and innovative strategy for continuing to build strong relationships" with such like-minded groups.
Chapman said the study committee does not envision a formal alternative organization to the BWA, but rather a looser network for using SBC resources to create a "teaching and learning" environment at the invitation of "conservative evangelical" Baptists in other parts of the world.
"In the long run [this] will be a much stronger contribution to our witness around the world," Chapman said.
Despite the SBC's long record of BWA membership and support, Chapman said: "In this generation--knowing that we've been a part of the Baptist World Alliance 99 years--in this generation--circumstances being as they are--does the BWA best represent who we are to Christians around the world, or has the time come when Southern Baptists can best represent themselves?"
Two Executive Committee members spoke in opposition to the recommendation.
Janet Hoffman, a pastor's wife from Bernice, La., serves by virtue of her office as national president of SBC-auxiliary Woman's Missionary Union. She took several minutes to describe a two-hour process of prayer and dialogue that culminated with a vote by the WMU executive board to continue their commitment to the BWA Women's Department despite the convention's withdrawal.
"I wish you could have been there, because it was as if they stood as one," he said. "Not one sound made, and as I looked in their faces, tears streaming down their faces, it was love that I saw there. It was not anything that was in reaction to anybody. It was love for their sisters."
Nancy McGuigan, a homemaker and school library assistant from Coatesville, Pa., read a resolution that she said was adopted unanimously the day before by an administrative committee of the Baptist Convention of Pennsylvania/South Jersey executive board. The resolution cited the regional convention's diverse ethnic makeup and urged the Executive Committee and BWA "to begin immediately to find a way for reconciliation for the good of the Kingdom." It carried the names of each member of the committee. "And I would like to add my name to the list," she said.
Others suggested delaying the vote until June, to see if one more meeting with BWA leaders might change anything.
But Mike Trammell, senior pastor of Mt. Airy Baptist Church in Maryland, urged passage of the recommendations by reciting names of the individuals who composed the study committee: Morris Chapman, Jimmy Draper, Tom Elliff, Paige Patterson, Paul Pressler, Jerry Rankin, Joe Reynolds, Gary Smith and Bob Sorrell.
"This is as fine and representative committee of Southern Baptists as we've ever had," Trammell said. "We need to trust these fine Southern Baptists we have empowered to do this task."
BWA General Secretary Denton Lotz told reporters immediately after the vote that he was "very sad" about the decision. He said he doesn't understand why Southern Baptists don't want to identify with Baptists who have struggled because of their beliefs in places like the former Soviet Union, South Africa, India and Bangladesh. Still, he added: "We live in hope. There are still 210 Baptist unions in the BWA."
Lotz said Southern Baptists are still welcome at BWA meetings and will continue to be included on BWA commissions. Asked how losing the SBC would affect BWA finances, Lotz said: "The Lord gave us a surplus this year. This is not a question of money. This is a question of fellowship."
Bob Casey, a retired physician from Florida who held a seven-day fast and vigil to protest the proposed BWA pullout, said he had been promised three minutes to speak at the meeting. But Executive Committee chairman Gary Smith said at the opening of the discussion that in the interest of giving members of the committee priority, he would not recognize guests from the gallery.
"Southern Baptists have a forum for any Baptist who is a messenger, and that is our convention," said Smith, pastor of Fielder Road Baptist Church in Arlington, Texas. "If you want to discuss this issue, I encourage you to come to the Southern Baptist Convention in Indianapolis."
Casey said he was disappointed that he wasn't allowed to speak. "I did everything they asked me to do," he said.
Bob Allen is managing editor of EthicsDaily.com.
See previous related stories:
Is CBF the Center of SBC Complaint Against BWA, Or Scapegoat?
SBC Vote on BWA Withdrawal Scheduled Tuesday
Floridian Marching to Protest Proposed BWA Pullout by SBC
Former BWA Leader Labels SBC Fundamentalism 'Heresy'
SBC Leader Says BWA is Obsolete
WMU Executive Board Affirms Ties With BWA
Questionable Enterprises?
BWA Leader Requests Delay in SBC Withdrawal
Editorials Critical of SBC/BWA Schism
Latin American Baptist Leaders Protest SBC Pullout of BWA
German Professor Denies SBC Study Committee Account of Comment
International Baptist Leaders Express Shock at SBC Plan to Leave BWA
SBC Recommends Defunding BWA
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text: Italian rock band Mod+á was founded when singer Francesco "Kekko" Silvestre met guitarist Matteo "Tino" Alberti and Enrico Palmosi (who later became their arranger). Soon, Paolo Bovi, an old acquaintance of Silvestre's, came in on keyboard, and the group began playing around Milan, the city from which they all hailed. Lead guitarist Diego Arrigoni, bassist Stefano Forcella, and drummer Manuel Signoretto joined, and before long, thanks to their determination to play as many shows... Read more


text:
GO Series: 10 Second Run DS
- Publisher: Gamebridge
- Release Date: Sep 20, 2010
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Jan 22, 2012For a $2 game you get a decent amount of content and a great concept to go with it. It's a nice game to kill some time away. It isn't spectacular or mind-blowing but that's expected out of a game like this
10 Second Run is very simple in every aspect – it looks simple, it sounds simple, and it plays simple. Also priced accordingly at just 200 Points, this is pretty much the best DSiWare game available for when you're making a short trip somewhere as you can achieve a lot in mere seconds.
This is platforming to the minimal expression. It has only ten seconds to show its virtues, and contains fun for hours.
The fast-paced, plentiful content will occupy even the most hardened gamers for hours with its extreme but just-about-feasible level of challenge, and at the price it should be nigh-on impossible to resist.


text:
Online Learning Readiness Questionnaire
'Before enrolling in an online course, you should first assess your readiness for stepping into the online learning environment. Your answers to the following questions will help you determine what you need to do to succeed at online learning. Post-survey feedback will also provide you with information on what you can expect from an online course.'
Material Type: Quiz/Test
Technical Format: HTML/Text
Date Added to MERLOT: February 12, 2013
Date Modified in MERLOT: February 20, 2017
Submitter: Cathy Swift
Keywords: online readiness of students
Reviewed by members of Editorial board for inclusion in MERLOT.Editor Review (not reviewed)
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Pick a Bookmark Collection or Course ePortfolio to put this material in or scroll to the bottom to create a new Bookmark CollectionName the Bookmark Collection to represent the materials you will addDescribe the Bookmark Collection so other MERLOT users will know what it contains and if it has value for their work or teaching. Other users can copy your Bookmark Collection to their own profile and modify it to save time.Edit the information about the material in this {0}Submitting Bookmarks...Select this link to open drop down to add material Online Learning Readiness Questionnaire to your Bookmark Collection or Course ePortfolioSelect this link to close drop down of your Bookmark Collection or Course ePortfolio for material Online Learning Readiness QuestionnaireSelect this link to open drop down to add material Online Learning Readiness Questionnaire to your Bookmark Collection or Course ePortfolio
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text:
Pumpkin Pie
Ever since the first pumpkin pie was served at the second Thanksgiving in 1623, this spicy custard pie has been a traditional American holiday treat.
Prep time: 45 minutes
Cooking time: 40 minutes
Cooling time: 1 hour
You Will Need
1 recipe single piecrust
1-1/2 cups cooked fresh or canned pumpkin purée
3/4 cup evaporated whole milk
2 large egg yolks
1/2 cup firmly packed dark brown sugar
1/4 cup maple syrup
2 tablespoons unsalted butter or margarine, at room temperature
2 teaspoons pumpkin pie spice
3 large egg whites
What to Do
1. Preheat the oven to 375°F. Make the piecrust fit into a 9-inch pie plate, and make a rope edge.
TIP: For a rope edge, pinch the pastry edge between the thumb and knuckle of your index finger at a 45° angle. Repeat all the way around, placing your thumb each time in the indentation just left by your knuckle.
2. In a large bowl, with an electric mixer on medium, beat the pumpkin purée, milk, egg yolks, sugar, syrup, butter and pumpkin pie spice. In a clean medium-size bowl, with clean beaters and the mixer on high, beat the egg whites until stiff but not dry. Fold the egg whites into the pumpkin mixture. Pour into the crust.
TIP: For sweet potato pie, substitute 1 1/2 cups fresh, cooked, or canned sweet potato purée for the pumpkin.
3. Bake for 40 minutes or until a knife inserted in the center comes out clean. Cool on a rack for 1 hour before serving.
Serves: 9
Per serving: Calories 345, Saturated Fat 8 g, Total Fat 17 g, Protein 7 g, Carbohydrates 44 g, Fiber 1 g, Sodium 88 mg, Cholesterol 74 mg
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A: A mechanic.


text:
Album: My Beautiful Dark Twisted Fantasy
Producer: Kanye West, DJ Frank E
Label: Roc-A-Fella, Def Jam
Much of Kanye's catalog deals with love, loss, and the perils of relationships in a celebrity world. "Blame Game" finds both John Legend and Kanye carrying on about dysfunctional relationships. Despite being able to bag any chick on the planet, Kanye has love issues just like the average joe, and he explained them in explicit detail on "Blame Game."
Chris Rock's "Yeezy taught me" bit at the end—no doubt aimed at Amber Rose—took on a life of its own, and will go down as one of the greatest rap catchphrases of all time. It all goes to show that dating someone like Kanye can change your life forever—for better or for worse.


text:
Logo modification job
This project was awarded to Djdesign for $35 USD.Get free quotes for a project like this
Project Budget$30 - $250 USD
Total Bids27
Project Description
I've a logo that need to re-create in illustrator format and make some modification.
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text:
Adds a button (via iframe) which easily lets your visitors Like your posts. Edit settings.
Currently that functionality has been disabled. It was producing an error and I will release a fix with the next version.
Just use the function
echo ahsfl_generate_button() and insert it anywhere in your WordPress theme files.
Whichever pages you want to not have the like button, give them the custom field "facebooklikebutton" with value "exclude".
Yes! You can call the function
ahsfl_get_image() elsewhere in your theme. It has one required argument (the post id) and one optional argument (the size, which defaults to 'thumbnail').
Requires: 2.7.2 or higher
Compatible up to: 3.0.5
Last Updated: 6 years ago
Active Installs: 300+
Got something to say? Need help?


text:
Cory Metcalf (Talk | contribs)
(→{{maxword|name=jit.gl.camera}} Tips and Workarounds)
Cory Metcalf (Talk | contribs)
(→People who looked at/used this object also looked at....)
|Line 101:
|Line 101:
==People who looked at/used this object also looked at....==
==People who looked at/used this object also looked at....==
==Errors or Clarifications==
==Errors or Clarifications==
The jit.gl.camera object allows you to define an OpenGL view. The view can either be directly rendered to an OpenGL drawing context or can be captured to a texture for further use.
With the help of jit.gl.node, it is easy to use multiple jit.gl.camera objects to view an OpenGL scene from multiple perspectives at once. In addition to multiple viewing positions and angles, each camera can have unique attributes for how it views the scene, such as field of view and clipping planes.
This patch uses four cameras to view all sides of a simple environment rendered to a jit.gl.node, then passes each cameras view out to be rendered on separate video planes:
You can find several interesting patches that make use of jit.gl.camera in the example patches included with Max:
Maxobjects.com listing for the jit.gl.camera object
(please list things that you believe to be errors or omissions from the existing refpage)


text:
1 Taylors Way,Hollis NH, 03049
4 beds , 3 Full/1 Half baths
3,300 sq. ft
Status: Active
Listing Provided Courtesy Of: BHHS Verani Nashua - Kathleen DeMello
Last Updated: 2/27/2017 7:19 AM
Description for 1 Taylors Way Hollis, NH 03049
Rare to market, this magnificent Reed's Farm home, sitting high on a ridge, boasts 4 bedrooms, (Master suite on 1st floor!) 3 full baths and a 1/2 bath, soaring ceilings, huge walk-in closets, expansive kitchen with tons of storage and eat-in area, shiny hardwood floors, formal dining area, a study, and 4 season sun room. If that isn't enough the full walk out basement is ready to be finished for your growing family. Sit on your deck and enjoy the ever changing views to Overlook Golf Course and the Nashua River. Want to go on vacation? No worries. This is a Home Owner's Association so all the landscaping, snow plowing, irrigation is taken care of for you. It's a life style you'll love living!
Listing Information
Public Water Service, Gas Water Heater
Public Sewer, Septic
Other, 200 Amps, Propane, Underground
Room Information
Kitchen, Living Room, Office
Master Suite
Interior Features
3,300 Sq. Ft. Above Ground
Hardwood, Ceramic Tile, Carpet
Central Air, Ceiling Fan(s)
Gas Heat, Forced Air
Walkout, Concrete, Daylight, Full, Entrance Inside, Unfinished Basement
Security System
Window Blinds, Cathedral/vaulted Ceiling, Gas Fireplace, Walk-in Closets
Exterior / Lot Features
2 Garage Spaces, Attached Garage, Paved Driveway, Finished Garage
Composition Shingle
Irrigation System, Landscaped, Other, Golf Course, Community Maintenance, Wooded
Financial Considerations
Inside AccessTM
Current market conditions for 1 Taylors Way
Active Market Trends
Source:MLS DATA
Median list price$612,400
Days on market122
Properties for sale34
Sold Market Trends
Source:MLS DATA
Median sold price$444,900
Sales to list price9,255.0%
Properties sold19
Median sq ft price$156
Arrows indicate 90 day trend
Listing Price History for 1 Taylors Way
Hollis Real Estate
Hollis is located in New Hampshire. Hollis, New Hampshire 03049 has a population of 7,679.
The median household income in Hollis, New Hampshire 03049 is $102,159. The median household income for the surrounding county is $70,472 compared to the national median of $53,046. The median age of people living in Hollis 03049 is 44.2 years.
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новости на русском
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Stagnation by design
06 February 2014 [17:30] -
By Joseph E. Stiglitz
Nobel laureate in economics, is University Professor at Columbia University. His most recent book is The Price of Inequality: How Today's Divided Society Endangers our Future.
Soon after the global financial crisis erupted in 2008, I warned that unless the right policies were adopted, Japanese-style malaise - slow growth and near-stagnant incomes for years to come - could set in. While leaders on both sides of the Atlantic claimed that they had learned the lessons of Japan, they promptly proceeded to repeat some of the same mistakes. Now, even a key former United States official, the economist Larry Summers, is warning of secular stagnation.
The basic point that I raised a half-decade ago was that, in a fundamental sense, the US economy was sick even before the crisis: it was only an asset-price bubble, created through lax regulation and low interest rates, that had made the economy seem robust. Beneath the surface, numerous problems were festering: growing inequality; an unmet need for structural reform (moving from a manufacturing-based economy to services and adapting to changing global comparative advantages); persistent global imbalances; and a financial system more attuned to speculating than to making investments that would create jobs, increase productivity, and redeploy surpluses to maximize social returns.
Policymakers' response to the crisis failed to address these issues; worse, it exacerbated some of them and created new ones - and not just in the US. The result has been increased indebtedness in many countries, as the collapse of GDP undermined government revenues. Moreover, underinvestment in both the public and private sector has created a generation of young people who have spent years idle and increasingly alienated at a point in their lives when they should have been honing their skills and increasing their productivity.
On both sides of the Atlantic, GDP is likely to grow considerably faster this year than in 2013. But, before leaders who embraced austerity policies open the champagne and toast themselves, they should examine where we are and consider the near-irreparable damage that these policies have caused.
Every downturn eventually comes to an end. The mark of a good policy is that it succeeds in making the downturn shallower and shorter than it otherwise would have been. The mark of the austerity policies that many governments embraced is that they made the downturn far deeper and longer than was necessary, with long-lasting consequences.
Real (inflation-adjusted) GDP per capita is lower in most of the North Atlantic than it was in 2007; in Greece, the economy has shrunk by an estimated 23%. Germany, the top-performing European country, has recorded miserly 0.7% average annual growth over the last six years. The US economy is still roughly 15% smaller than it would have been had growth continued even on the moderate pre-crisis trajectory.
But even these numbers do not tell the full story of how bad things are, because GDP is not a good measure of success. Far more relevant is what is happening to household incomes. Median real income in the US is below its level in 1989, a quarter-century ago; median income for full-time male workers is lower now than it was more than 40 years ago.
Some, like the economist Robert Gordon, have suggested that we should adjust to a new reality in which long-term productivity growth will be significantly below what it has been over the past century. Given economists' miserable record - reflected in the run-up to the crisis - for even three-year predictions, no one should have much confidence in a crystal ball that forecasts decades into the future. But this much seems clear: unless government policies change, we are in for a long period of disappointment.
Markets are not self-correcting. The underlying fundamental problems that I outlined earlier could get worse - and many are. Inequality leads to weak demand; widening inequality weakens demand even more; and, in most countries, including the US, the crisis has only worsened inequality.
The trade surpluses of northern Europe have increased, even as China's have moderated. Most important, markets have never been very good at achieving structural transformations quickly on their own; the transition from agriculture to manufacturing, for example, was anything but smooth; on the contrary, it was accompanied by significant social dislocation and the Great Depression.
This time is no different, but in some ways it could be worse: the sectors that should be growing, reflecting the needs and desires of citizens, are services like education and health, which traditionally have been publicly financed, and for good reason. But, rather than government facilitating the transition, austerity is inhibiting it.
Malaise is better than a recession, and a recession is better than a depression. But the difficulties that we are facing now are not the result of the inexorable laws of economics, to which we simply must adjust, as we would to a natural disaster, like an earthquake or tsunami. They are not even a kind of penance that we have to pay for past sins - though, to be sure, the neoliberal policies that have prevailed for the past three decades have much to do with our current predicament.
Instead, our current difficulties are the result of flawed policies. There are alternatives. But we will not find them in the self-satisfied complacency of the elites, whose incomes and stock portfolios are once again soaring. Only some people, it seems, must adjust to a permanently lower standard of living. Unfortunately, those people happen to be most people.
Copyright: Project Syndicate
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text:
Yes, that's Betty without braces, without glasses and without polka dots. In the next episode of "Ugly Betty," we see what life would be like if Betty weren't ugly.
In the sneak peek we posted earlier this week, we saw that Betty's braces aren't going to come off without some hitches.
And while Kathy Najimy guest stars as Betty's orthodontist, she also plays Betty's ghost of the beautiful life.
In that alternate world, Betty is in charge, fashionable and, from what we can tell – Wilhelmina-ish.
Check out the sneak peek videos below:
What did you think of "Pretty Betty"?
These videos are set to expire on March 25.
Follow Zap2it on Twitter, @TheRealJethro on Twitter, and Zap2it on Facebook for all your movies, TV and celebrity news.
'Ugly Betty' sneak peek: Kathy Najimy attacks Betty's braces
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All 'Ugly Betty' show recaps
Photo credit: ABC


text:
Interlinking Sites: Is It A Good Idea?
With Google's constant algorithm updates it's no wonder webmasters are concerned about link building. What works today may be disallowed tomorrow and your whole site could come tumbling down around your ears. Should you interlink your own blogs and sites? Absolutely. As long as you follow a few important guidelines.
Subscribe to my exclusive newsletter, and learn secret SEO tactics to boost your traffic»
Google looks at the number of incoming links you have, as well as the quality and relevancy of each link. The more "good" links you have coming into your blog, the higher your authority. On the other hand, too many "bad" links and you're doomed. What's the difference? Let's take a look at the Google Webmaster Guidelines.
Regarding linking between your own sites, the Google Guidelines state that "building partner pages exclusively for the sake of cross-linking" will have a negative impact on your site's ranking.
However, the keyword in that sentence is "exclusively." It's been common practice to build Squidoo lenses or a Weebly blogs, or other off-site properties, throw in some shoddy content, and use those properties to link back to your main blog or website. Those off-site properties were used exclusively as a means to build links.
These days, if you want to use those same techniques, you have to use good content. If you build a Squidoo lens or a HubPage it has to be something that other bloggers would feel comfortable linking out to as a credible resource.
Now, before you think it's too much work to create killer content just to carry a link, remember why you're doing it. Those off-site properties often come with a lot more link authority than your blog has. Your blog might be a PR2 and Squidoo is a PR8. That link coming from Squidoo will give your site quite a boost, so it's well worth the effort to create quality content.
Quality isn't the only criteria, though, that content must also be relevant.
It's easier to understand if you look at it from Google's viewpoint. They want you to create quality content that generates its own natural links, content that other bloggers would want to link to. Which means the content on either end of the link must be relevant. Another blogger writing about African Elephants wouldn't likely link to your blog post about Travel in Africa, and you shouldn't either.
Your blogs may all be in closely related niches but that's not good enough, especially if you're just building those blogs so they can be used exclusively as a means to build links. If you want to interlink between your own sites, link from post to post, or page to page. Not from Homepage to Homepage. That way you're linking between relevant content.
If you want to avoid even the possibility of a problem, forget about building your own links altogether and focus on creating the kind of content that generates natural links. Ideally, that's what Google's looking for. And why you may be able to get by the bots with these link building techniques now, who knows what changes Google will make in the future. Go ahead and do some interlinking between your own blogs and sites – where it's appropriate – but focus on getting those natural links.
When you're learning search engine optimization, you need to understand that the very first step is to create a website. If you're interested in starting your own blog, I have written a step-by-step guide that will show you how to start blogging for money for as little as $3.49 per month (this low price is guaranteed only through my link). You will also receive your own domain name for free ($15 value) by clicking on this link and purchasing at least 12 months of hosting with BlueHost. Keep in mind that if you're learning how to optimize your website, the first thing you need is your own self-hosted website. It will help you look more professional in front of your visitors, clients, companies, and everyone else, including search engines.


text:
Judaism: The First Phase Paperback
The Place of Ezra and Nehemiah in the Origins of Judaism
by Joseph Blenkinsopp;
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Judaism: The First Phase
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Most studies of how early Judaism related to the non-Jewish world and how it was perceived by others start no earlier than the Hellenistic period. Joseph Blenkinsopp argues that we must go further back, to the Babylonian destruction of Jerusalem and its temple and the liquidation of the political and religious infrastructure monarchy, priesthood, scribalism, prophecy which had sustained the Judean state for centuries. / Moving beyond the ideologically driven approaches of scholars over the past two centuries, he explores such pragmatic issues as the emergence of a distinctive group identity in the aftermath of the fall of the Judean state, the degree of continuity-discontinuity between national identity before the exile and competition among distinct group for legitimacy after it, and the historical realities behind the idea of a restoration in a fundamentally different world, with neither monarchy nor statehood and a much-diminished temple. / Judaism, the First Phase is a fresh and potentially stunning look at Jewish origins, tracing the legacy of Ezra and Nehemiah. Ideal for scholars and students."
Judaism: The First Phase by Joseph Blenkinsopp was published by Eerdmans Publishing Company in September 2009. The ISBN for Judaism: The First Phase is 9780802864505.
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Details for Judaism: The First Phase
|Page last updated
|1st September 2016
|Author / Artist
|Joseph Blenkinsopp
|Book Format
|Paperback / softback
|Eerdmans Publishing Company (September 2009)
|Number of Pages
|25 mm
|226 mm
|150 mm
|Product Code


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|"Xavier Hanin" <xavier.ha...@gmail.com>
|Re: useOrigin and type filtering
|Tue, 06 Mar 2007 13:11:20 GMT
On 3/6/07, Stephane Bailliez <firstname.lastname@example.com> wrote: > > I'm messing with useOrigin and the culprit is actually completely lost > in getting me the correct artifacts when using cachepath. > I have artifacts with different types for the same conf(ie: jar, source, > javadoc) and while it works fine when doing a useOrigin=false with using > useOrigin=true it manages to push me artifacts of type source instead of > jar, so some filtering is going berserk somewhere. > > <ivy:resolve useorigin="${ivy.useorigin}" conf="*" showprogress="false" > type="jar,javadoc,source"/> > <ivy:cachepath useorigin="${ivy.useorigin}" > pathid="ivy.compile.classpath" type="jar" conf="compile"/> > > I haven't yet investigated the problem, but ouf of curiosity, has anyone > seeing this behavior before ? > (ie: I'm using 1.4.1) No, at least not yet :-) And I'm surprised of this problem, because I see no relation between the two. Except if the bug is that when using origin the artifact type is lost... Strange. Side note: there should be absolutely no need whatsoever to duplicate > useorigin in every task once it has been resolved. I agree it is weird, you can open an issue and relate it to scoping. - Xavier This is just a killer > for modularity as if I want to enable this feature, I have to go through > all build files that use retrieve to do some special packaging or > layout. This information should be self contained in the resolution > data. Scoping should hopefully help to get rid of this. > > -- stephane >
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(Played by James Cromwell [1996] and Glenn Corbett [1967]) The renowned Terran American scientist and developer of warp drive on his world (AD 2030-2117?), an eccentric genius who became a...
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(Played by Michael Dorn) PERSONNEL FILE: Worf Last Starfleet Rank: Lieutenant Commander Current position: Federation Ambassador to Qo'noS; previously, strategic operations...
Picard, Jean-Luc
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Starfleet Engineer aboard the U.S.S. Enterprise-E in 2373. When the Borg began assimilating the ship, Porter was among those added to the Collective.
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In the "Dixon Hill" stories favored by Captain Jean-Luc Picard as a form of holodeck entertainment, Ruby was one of several rich characters who had an connection to Dixon.
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A member of the Zefram Cochrane engineering team that constructed the first warp drive in 2063. Sloane also helped Picard defeat the Borg aboard the U.S.S. Enterprise NCC-1701-E.


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Eike Batista net worth: Eike Batista is a Brazilian businessman and mogul who has a net worth of -$1.2 billion. Between August 2012 and August 2013, Eike Batista's wealth has plummeted to a shocking degree. His net worth once peaked at $30 billion which made him the 10th richest person in the world and the number one richest person in South America. Eike Batista is the president and owner of EBX, a Brazilian conglomerate made up of several different companies (many of which include the letter "X" in their name). Batista is the son of famous Brazilian businessman Eliezer Batista, who moved Eike and his 6 siblings all around the world as part of his job. He went to school in Germany where he stayed even after his family returned to Brazil, reportedly making ends meet by selling insurance policies door to door! He eventually came back to Brazil, though, and chose to focus on lucrative trades such as gold and diamonds. At only 23 years old, Batista started his first company, a gold trading firm called "Autram Aurem." This young company had made an impressive $6 million after only a year and a half in operation. His father is Eliezer Batista da Silva, a longtime head of mining company Vale. Born in Brazil, he dropped out of college without completing his degrees. He was once married to Luma de Oliveira a famous Brazilian actress, Playboy cover girl, and carnival queen, the couple of 2 children. Batista is also known for his high profile on the internet. He has especially taken to the social networking and microblogging site Twitter, on which he is very active and has around 1.3 million followers. He uses this platform to discuss his numerous companies and communicate with other Twitter users.
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Eike Batista
Date of Birth
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Entrepreneur, Businessperson
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This review is from: Intel Core i7-2700K Sandy Bridge Quad-Core 3.5GHz (3.9GHz Turbo) LGA 1155 95W BX80623i72700K Desktop Processor Intel HD Graphics 3000
Pros: Runs all my games with no issues. Easily manages anything I have tested so far.
Cons: The heat sink it comes with is really cheap, but I was already going to use a different one.READ FULL REVIEW
This review is from: ASUS SABERTOOTH Z77 LGA 1155 Intel Z77 HDMI SATA 6Gb/s USB 3.0 ATX Intel Motherboard
Pros: Easy to install, and a lot of nice features for overclocking.
Cons: I dont like where the USB 3.0 is placed, but that may just be the case I bought.READ FULL REVIEW


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- 5 years limited
Shell 10/24/2013
This review is from: SanDisk Cruzer Edge 2GB USB 2.0 Flash Drive 128bit AES Encryption Model SDCZ51-002G-B35
Good performance. I like the password to access protecting and securing files. Compact and retractable plug.
Weak Shell Design. One of the most important features to prevent loss of the unit and your data is where the unit fails and why I can only give it 2 eggs.
The key ring hole on the end somehow cracked on mine in the middle which allows it to slip off the key ring is any force is applied . I have to take extra measures just to make sure I don't lose the unit. I can no longer carry it on my key ring. I have to keep it in my backpack instead.
The retractable plug extends when you don't want it to. Have to constantly re-stow it.
Other Thoughts:
I would not recommend this product for anyone who intends to keep it on a key ring or lanyard.
Manufacturer Response:
Hello User,
My name is Allen from SanDisk support. I am sorry to learn that key ring hole of the flash drive has broken. I would suggest you to please contact SanDisk support at 1-866-SANDISK or submit a support request at www.sandisk.com/support for further assistance.
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Shah Residency
31/A, N.H. pani house, Lal Market, Gangtok 737101, India
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post #1 of 4 Old 10-13-2006, 12:01 AM Thread Starter
Spelling tickles the mind...
I cdnuolt blveiee taht I cluod aulaclty uesdnatnrd waht I was rdanieg. The phaonmneal pweor of the hmuan mnid. Aoccdrnig to a rscheearch at Cmabrigde Uinervtisy, it deosn't mttaer in waht oredr the ltteers in a wrod are, the olny iprmoatnt tihng is taht the frist and lsat ltteer be in the rghit pclae. The rset can be a taotl mses and you can sitll raed it wouthit a porbelm. Tihs is bcuseae the huamn mnid deos not raed ervey lteter by istlef, but the wrod as a wlohe. Amzanig huh? yaeh and I awlyas thought slpeling was ipmorantt! tahts so cool!!!
Read..read..read..Get it? You can understand...not..
Read..read..read..Get it? You can understand...not..
post #2 of 4 Old 10-13-2006, 01:04 AM
Wow..... I actaully udonsterood it.
post #3 of 4 Old 10-13-2006, 01:07 AM Thread Starter
Criatenly it deos.
I like the way the researcher manages to scramble the letters. If only I write the way she does. Then people will be surprised to death by the mess.
I like the way the researcher manages to scramble the letters. If only I write the way she does. Then people will be surprised to death by the mess.
post #4 of 4 Old 10-13-2006, 01:56 AM
lol, i could read it, also, and spelling is one of my strengths, go figure :P
You know what the definition of hobby is: something of interest that is going to cost you lots of money!
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The Badland 2500 lb. ATV/Utility Electric Winch with Wireless Remote Control from Harbor Freight Tools is a must-have for off-roading adventure and can also be used for loading a boat. A remote switch makes this winch easy to use even when you're alone.
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2500 lb. ATV/Utility Electric Winch with Wireless Remote Control


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Lee Evans is an English comedian, writer, actor and musician who has a net worth of $8 million. Lee Evans has earned his net worth from his 1990's career as a loud, hot, sweaty, energetic stage performer and from his physical observational comedy routine. Lee was born on February 25, 1946 in Avonmouth, Bristol, England. His mother was Irish and his father Dave Evans was a Welsh nightclub performer. He attended The Billericay School in Billericay, Essex and then he spent two years at Thurrock Art College in Essex as a boxer. He also attended Thurrock Art College in Essex. As a teenager Lee played drums in the punk rock named The Forgotten Five. He has been married to Heather Nudds since 1984 they have one daughter and currently lives in Billericay, England. He won the Perrier Comedy Award for his work at the Edinburgh Festival in 1993.Evans has been credited with breaking the world record for performing in front of the largest audience of 10,108 people watched his solo act at the Manchester Evening News Arena in November of 2005.His 2008 UK tour entitled "Big" included 59 dates and was performed in front of over 500,000 people. On March 30th, 2010 Evans was part of Channel 4's Comedy Gala, this event was to raise money for Great Ormond Street Hospital, and Evans received a comedy award for his performance and promptly auctioned it off to the audience. Some of his most recent performances include another UK tour in 2011 entitled "Roadrunner" this show was booked for 50 dates from August to November.
Lee Evans Net Worth
Lee Evans
Date of Birth
Feb 25, 1964 (53 years old)
Musician, Actor, Screenwriter, Stand-up comedian, Film Score Composer
United Kingdom
5 ft 8 in (1.75 m)
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All net worths are calculated by applying a proprietary algorithm. The results are then fact checked and confirmed by a team of editors. We scan all publicly available data and resources and work diligently to ensure that our numbers are the most accurate and up to date net worth data you will find anywhere on the internet.