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Microvascular networks undergo patterning changes that determine and reflect functional adaptations during tissue remodeling. Alterations in network architectures are a result of complex and integrated signaling events. To understand how two growth factor signals interact to stimulate angiogenesis and arterialization, we engineered spatially directed microvascular pattern changes in vivo by using combinations of focally delivered exogenous growth factors. We implanted microdelivery beads containing recombinant vascular endothelial growth factor-164 (VEGF164) and recombinant angiopoietin-1* (Ang-1*) into the dorsal subcutaneous tissue of fully anesthetized male Fischer 344 rats implanted with backpack window chambers, and we quantified vascular patterning changes by using intravital microscopy, a combination of architectural metrics, and immunohistochemistry. Focal delivery of VEGF164 caused spatially directed increases in both the total number and the density of vessels with diameters <25 μm 7 days after microbead implantation. Increases were maintained out to 14 days but were reduced to control values by day 21. The addition of Ang-1* on day 7 maintained these increases out to day 21, induced vessel order ratios comparable to control levels, and was accompanied by increases in the length density of smooth muscle α-actin-positive vessels. We achieved spatial control of patterning changes in vivo by using multisignal stimulation via focal delivery of exogenous growth factor combinations and conclude that Ang-1* administered subsequent to VEGF164 stimulation induces vascular growth while maintaining a network pattern consistent with native patterns that persist in the presence of vehicle control stimulation.
- microcirculation
- window chamber
- microvascular remodeling
- arteriogenesis
- vascular tissue engineering
- vascular endothelial growth factor-164
- angiopoietin-1
microvascular remodeling in the adult animal underlies adaptive and regulatory events in normal life, such as wound repair (19, 43) and exercise-induced skeletal muscle adaptation (9, 22, 46), as well as in pathological conditions, such as myocardial responses to atherosclerosis (5) and tumor growth (17, 26). This complex process involves angiogenesis (20) or capillary formation from an existing capillary bed, arterialization (6, 33, 40) involving the recruitment and differentiation of contractile perivascular cells, and vessel regression (1, 3, 4, 7). Although epigenetic stimuli may include changes in hemodynamic stresses that impinge on the vessel wall, coordination of these events is ultimately achieved by biochemical signals, such as growth factor proteins. For example, vascular endothelial growth factor (VEGF) mediates angiogenesis by promoting endothelial cell recruitment, proliferation, and tube formation (14, 17, 21), whereas angiopoietin-1 (Ang-1) is thought to promote vessel stabilization and maturation (2, 42).
When a network is structurally remodeled, the architecture, or pattern, of the microvascular bed is altered. The pattern is suggestive of the function served by the particular network, the extent to which it is served, and the stimuli through which the pattern was created (24, 37, 38). Therefore, pattern formation, the assembly of blood vessels into a functional network of arterioles, capillaries, and venules of particular length and diameter distributions, is a central component of microvascular remodeling. Studying induced pattern changes can improve the understanding of how environmental stimuli orchestrate the remodeling process.
Determining the spatial and temporal effects of specific growth factors on microvascular pattern formation also has far-reaching therapeutic possibilities and tissue engineering applications. For example, a thorough understanding of the growth factors implicated in microvascular remodeling combined with their controlled delivery can direct the assembly of unique, tissue-specific networks for collateralization therapy in the rescue and healing of ischemic tissues. Knowledge of the growth factors' roles in pattern formation is critical to engineering tissues that require an adequate blood supply. The objectives of this study are to understand how multisignal combinations of angiogenic growth factors that have previously been shown to have associated roles in microvascular remodeling (2, 42, 44) work in concert to influence network patterning during remodeling and to evaluate whether these factors can spatially and temporally direct angiogenesis and arterialization to focal regions of tissue. We spatially manipulated network patterning in vivo by using focal exogenous delivery of recombinant VEGF164, Ang-1, and combinations of these growth factors, and we studied the ensuing localized patterning changes to obtain a signature of growth factor effects at the network level.
Window chamber apparatus. All experiments were performed using sterile techniques in accordance with the guidelines of the University of Virginia Animal Care and Use Committee. The dorsal skinfold backpack window chamber used in this study (32) allows for the noninvasive intravital observation of a naturally vascularized in vivo tissue throughout a 21-day period (Fig. 1). One hundred and twenty-one 250- to 300-g male Fischer 344 rats were anesthetized with intramuscular injections of ketamine (80 mg/kg body wt), atropine (0.08 mg/kg body wt), and xylazine (8 mg/kg body wt). Dorsal skin was folded along a centerline over the spine and suspended by the chamber. Epidermis and dermis on both sides of the chamber were surgically dissected to expose vascularized subcutaneous tissue, and protective glass shields were placed over the tissue. The total window area enclosed by the backpack chamber and protected from the environment by glass coverslips was 0.785 cm2, and the average thickness of the window tissue was ∼150 μm. After surgery, rats were placed in heated cages and returned to the vivarium. While in their cages, rats were given rat chow and water ad libidum.
Noninvasive intravital image acquisition. Images of window chamber tissues were obtained noninvasively. At each time point, rats were anesthetized as described in Window chamber apparatus, glass cover-slips were removed, and the tissues were flushed with 0.9% NaCl sterile saline. Tissue was always imaged from one side of the chamber, selected based on image clarity on day 1. With the use of a ×20 dry objective, images of nine adjacent fields of view were captured and assembled into a single rectangular montaged image measuring 0.09 mm × 1.20 mm that was centered on the alginate bead. Total area imaged per window quadrant was 1.08 mm2, and all of this area was contained within a 1-mm radius of the alginate bead delivery device. Images were also recorded and these recordings were subsequently used during the data collection to indicate flow directions in the network. Visualization of vessels was enhanced by using a 443-nm bandpass hemoglobin optical filter. During measurement acquisition, steps were taken to maintain constant ambient temperature; measurements were obtained in a thermostated room at 22°C.
Tissue harvest, fixation, and histology. Rats were anesthetized and entire window tissues were excised, whole mounted on gelatin-coated glass slides, and fixed in 4% paraformaldehyde overnight in —4°C. Tissues were immunolabeled for smooth muscle α-actin (SMA) using purified Cy3 conjugated clone 1A4 mouse monoclonal anti-SMA (Sigma Biosciences, St. Louis, MO) diluted 1:400 in phosphate-buffered saline containing 0.1% saponin and 2% bovine albumin (Fischer Scientific, Pittsburgh, PA) at pH 7.4 (incubation for 1 h at room temperature). One field of view per window quadrant (area equal to 1.08 mm2) was examined under a ×10 dry objective using a Nikon TE-300 inverted microscope with confocal accessories (Bio-Rad μ-Radiance). Images were digitized and analyzed by using Scion Image software version 4.0.2 (Scion, Frederick, MD), wich was calibrated to measure the length of positive SMA staining along the vessels.
Microvascular network patterning measurements. Functional length density, diameter distribution, and vessel order ratios were used to describe patterning changes and identify the different types of network architectural adaptations. The Strahler ordering scheme (13, 41) was used to classify vessels into different orders. Capillaries, identified under the light microscope based on recognition of their characteristic single-file line of red blood cell flow, were designated as order 1 vessels, and sequential orders were assigned progressively upstream incrementing by branch points at which two vessels of the same order joined. Arteriovenous shunts were designated with an order value equal to 1 less than that of the upstream vessel. Arteriole anastomoses were rare, but when seen, were designated with the order value equal to one greater than the immediate downstream vessel. This ordering scheme for vessel loops preserved the downstream-to-upstream-oriented notation while capturing connectivity changes. Vessel order ratios were computed by dividing the total number of vessels in one order (n) by the total number of vessels in the order succeeding it (n + 1).
Digitized intravital microscopy images were analyzed by using Scion Image software. Vessel segments were traced, and lengths (defined as distances between branch points) and internal diameters (based on blood column widths) were recorded. Order value was assigned to each vessel segment. Lengths of vessels containing SMA-expressing perivascular cells were obtained from immuno-stained histological whole mounts.
Growth factor diffusion characterization. To quantify the amount and spatial distribution of VEGF164 diffusing out of the alginate bead and into the tissue, VEGF164 was iodinated with 0.5 mCi of 125I (Sigma Chemical) by using commercially available IodoBeads according to the protocol described by the manufacturer (Pierce, Rockford, IL) and delivered either via filling the entire chamber volume (1.6 μg/ml) and then sealing the window chamber (superfusion over the entire tissue) or via placement of alginate beads containing 125I-labeled VEGF164, one into each of the four tissue quadrants. Sterile 0.9% NaCl saline solution was dripped over the windows, and glass coverslips were placed on top. The tissues were maintained at room temperature for 2, 23, 47, or 71 h and then exposed to radiographic film for 1 h. Film was digitized, and Scion Image software was used to measure grayscale intensity levels correlated to 125I-labeled VEGF164 concentrations within the tissue. Intensity measurements for 125I-labeled VEGF164 superfusion were obtained along eight equally spaced radial directions from the center of the window chamber outward to the chamber's periphery. Intensity measurements for the bead delivery studies were obtained and averaged along four perpendicular radial directions from the bead outward to a distance of 3 mm from the bead.
Growth factor delivery. Alginate bead delivery devices (39) were placed in 50 μl of 10 μg/ml growth factor; either rat recombinant VEGF164 (R&D Systems) or recombinant Ang-1 (Ang-1*) [containing a modified NH2-terminus (45), Regeneron] in 10 mM phosphate-buffered saline (pH 7.4, Sigma Chemical) overnight and implanted into window tissues of anesthetized rats, one per window quadrant (Fig. 1), 4 days after window chamber implantation surgery (denoted as day 1). Vehicle control was 10 mM phosphate-buffered saline. The 4-day delay allowed the animals to recover from surgery and provided a suitable time period for network equilibration, as determined by initial characterization studies. Therefore, day 1 designates the fifth day after window implantation surgery, and all subsequent time points in this study (up to day 21) are referenced accordingly. Alginate beads were held in stationary positions in the tissue by the natural adhesiveness of the hydrogel throughout the 21-day study. Window quadrants were not physically isolated from neighboring quadrants. One study group was subjected to single time point VEGF164 superfusion, which was delivered in 1.6 μg/ml over the entire tissue surface by filling the chamber volume and then sealing the window chamber. A uniform concentration was maintained over 72 h as determined by characterizing growth factor diffusion (data not shown) using the method described above.
Statistical analysis. All results are presented in the form of means ± SD. All comparisons were made by using the statistical analysis tools provided by SigmaPlot 5.0 (SPPS). Data were tested for normality and analyzed by one-way ANOVA, followed by nonpaired Tukey's t-test. Significance was asserted at P < 0.05.
Baseline remodeling characterization. To assess baseline remodeling in response to the window chamber implantation surgery, patterning metrics (±SD) in 12 animals were characterized after window chamber implantation (Fig. 2). Functional length densities were calculated in different diameter groupings (0–25, 26–50, 51–75, and 76–100 μm) (data not shown). The only significant difference (P < 0.05) in functional length density from day 1 was seen on day 3, during which the functional length density of the smallest diameter vessels (<25 μm) increased (Fig. 2A). By day 5 this value had returned to that seen on day 1 and remained statistically similar to values on day 1 throughout the remainder of the study. There was no significant difference in functional length density at any time compared with day 1 in any other diameter category (data not shown).
Diameter distributions were calculated as a percentage of the total number of vessels (arterioles, capillaries, and venules) in a particular diameter grouping. At day 3, the percentage of vessels in the smallest diameter group had increased compared with day 1, but this value returned to the initial values by day 5 and remained there (Fig. 2B). Furthermore, there was no statistically significant difference in any of the vessel order ratios at any time point compared with day 1 or in SMA coverage length at any time points compared with day 1 (data not shown).
Growth factor superfusion. We hypothesized that the patterning effects elicited by focal exogenous growth factor exposure, wherein a concentration gradient was delivered over a duration of 72 h, would be distinct from those elicited by a single time point, spatially homogeneous growth factor exposure, wherein the spatial gradient and time-release delivery was eliminated. To test this hypothesis, one study group received a single superfusion of the entire tissue surface of either vehicle control or VEGF164. The concentration of superfused VEGF164 (1.6 μg/ml) was approximately equal to that experienced by a region of tissue 0.5 mm from the alginate beads after 3 h of bead implantation. This concentration was selected for superfusion because it was relevant to subsequent VEGF164 alginate bead stimulation and fell within a range previously shown to invoke a significant architectural remodeling response (27). There were no significant differences in the average functional length density of vessels with diameters <25 μm, percentage of vessels in this diameter category, order ratios, or SMA coverage lengths between microvascular beds stimulated with vehicle control (delivered via superfusion or alginate beads) versus those treated with VEGF164 superfusion (data not shown).
Growth factor diffusion characterization and delivery vehicle placement. 125I-labeled VEGF164 is released from the alginate bead into the tissue gradually over a 72-h time period (Fig. 3A). All of the growth factor remains in the tissue within 1 mm of the alginate bead, and a concentration gradient exists at all four time points.
To test the spatial sensitivity of the delivery device in eliciting a localized response, we utilized a six-region bead implantation scheme (Fig. 3C), wherein alginate beads incubated in either 10 μg/ml VEGF164 or vehicle control (phosphate-buffered saline, pH 7.4) were placed into one of each of the six tissue regions in a hexagonal configuration. In this configuration VEGF164-bead spacing (2 mm apart) was identical to twice the maximum distance of growth factor diffusion (1 mm from the bead). We hypothesized that if the patterning response was limited to the region bounded by the VEGF164 diffusion, there would be comparable responses among the three regions stimulated by VEGF164-beads that would be distinct from the responses seen in regions stimulated by control beads, and we would have achieved spatially directed microvascular patterning.
Our hypothesis was confirmed by the data reported in Fig. 4. Tissue regions stimulated with microdelivery beads containing VEGF164 (regions 4, 5, and 6) were statistically similar to one another in all metrics, whereas control-treated tissue regions (regions 1, 2, and 3) were also similar to one another in all metrics at all time points. Stimulation with VEGF164-containing alginate beads produced nearly twofold increases in functional length density in the smallest diameter vessels (<25 μm) at two time points compared with control stimulation: days 7 and 14 (Fig. 4A). However, by day 21 functional length density in the VEGF164 bead-stimulated tissues was decreased to control bead-stimulated levels. Similarly, the percentage of vessels with diameters <25 μm was significantly increased at days 7 and 14 in the VEGF164 bead-stimulated tissue over that in the control bead-stimulated case, but by day 21 this percentage was reduced to the control level (Fig. 4B). Interestingly, the ratio values for 1:2 order vessels, reflecting the number of capillaries to the number of vessels located one branchpoint upstream, were also significantly increased at days 7 and 14 for the VEGF164 bead-stimulated tissue over those in the control bead-stimulated case, although this ratio returned to the control by day 21 (Fig. 4C). SMA coverage lengths in tissues stimulated with VEGF164, as a percentage of total vessel length, were comparable to control percentages at all time points throughout the study (Fig. 4D). Furthermore, VEGF164 bead-stimulated tissue did not differ significantly from the control treatment in functional length density, diameter distribution, order ratio, or SMA coverage at any time point in vessels larger than 25 μm (data not shown). Finally, the six-region bead insertion scheme generated similar results to those generated by using a four-region implantation scheme (Fig. 3B; data not shown), affirming the hypothesis that the network patterning changes were limited to the regions of tissue stimulated by the growth factor gradients.
Directed patterning changes stimulated with VEGF164 vs. Ang-1*. To study the effects of Ang-1* on microvascular network patterning compared with VEGF164, 12 rats implanted with dorsal skin backpack windows received alginate beads containing VEGF164 or Ang-1* and vehicle control, implanted one per tissue quadrant (Fig. 3B) on day 1. These interventions generated localized network patterning changes that were distinct and spatially limited to the quadrants containing the microdelivery device (Fig. 5).
On days 7 and 14, VEGF164 caused significantly increased functional length densities (Fig. 6A) and increased percentages of <25 μm diameter vessels (Fig. 6B) compared with stimulation with control beads at these time points. At no time point was there a significant difference in either of these metrics in tissue quadrants stimulated by Ang-1* compared with those stimulated with vehicle control. Ratio values of 1:2 order were increased in the VEGF164 bead-stimulated group over the control-treated group after 7 and 14 days (Fig. 6C), but there was no significant difference between the control and Ang-1*-stimulated groups at any time point for this order ratio. SMA coverage length in <25 μm diameter vessels at days 4 and 7 was significantly increased in the Ang-1*-stimulated group compared with the control bead-stimulated group (Fig. 6D), but no differences were seen in the VEGF164-stimulated group.
Directed patterning changes stimulated by sequential applications of VEGF164 and Ang-1*. To study how temporal association of VEGF164 and Ang-1* could direct localized angiogenesis and the perivascular cell recruitment process associated with arterialization, 36 rats were divided into three equal groups, and growth factor or control treatment was administered via alginate bead on day 1 and day 7 by removing the initial microdelivery bead and implanting the second bead in the same location. One group received VEGF164 followed by vehicle control, another group received VEGF164 followed by Ang-1*, and a third group received VEGF164 followed by VEGF164 treatment (Fig. 7). On days 7 and 14, length densities (Fig. 8A) and percentages of <25 μm diameter vessels (Fig. 8B) were significantly increased for VEGF164/VEGF164, VEGF164/Ang-1*, and VEGF164/control groups compared with control/control. For the VEGF164/VEGF164 and VEGF164/Ang-1* groups, functional length density remained significantly elevated compared with control-control at day 21. However, the length density of VEGF164/control-stimulated vessels in this diameter category returned to control/control levels.
There was a statistically significant increase in the 1:2 order ratio values of all treatment groups compared with control/control values on day 7 (Fig. 8C). On day 14, the VEGF164/VEGF164-stimulated tissue ratio and VEGF164/control-stimulated tissue ratio remained elevated above control levels; however, the 1:2 order ratio from VEGF164/Ang-1*-stimulated tissues had decreased to control levels. By day 21 there were no significant differences between growth factor-treated study groups and control/control-treated study group. The length of <25 μm diameter vessels coated with SMA-positive cells was significantly increased on day 14 in the VEGF164/Ang-1*-stimulated tissues compared with control-control and VEGF164-VEGF164-stimulated tissues. VEGF164/control and VEGF164/VEGF164 groups did not differ significantly from the control/control treatment group at any time point.
Previous investigations of microvascular remodeling have been focused on whole network adaptation in response to growth factor delivery by using large-scale exogenous applications of single angiogenic growth factors. In most of these studies the remodeling response was quantified by measuring the changes in total functional vascular density over time (10, 31). The current study uses a detailed pattern analysis approach to study microvascular remodeling patterning changes in response to spatially and temporally controlled focal applications of growth factors. In addition to capillary growth, vessel maturation via the acquisition of a perivascular cell coating, representing the early stages of the arterialization process, was also examined by using a combination of metrics that collectively describe pattern alterations. Unlike most previous studies, this research was conducted in healthy adult tissue, of which its natural response to wounding is microvascular remodeling. Consequently, the cells and signaling processes involved in microvascular remodeling are assumed to be both functional and naturally occurring. Microvascular network patterning is the result of many processes, including angiogenesis, arterialization, and regression, which are coordinated by multiple signaling events (33) and up to 1,000 genes (21). As molecular signals work in concert to elicit a cascade of cellular behaviors that contribute to this continuum, we developed experiments to examine the effects of stimulating remodeling using a temporal combination of VEGF164 and Ang-1*.
One main finding of this study is that temporally controlled focal applications of exogenous VEGF164 and Ang-1* administered via alginate microbeads spatially direct microvascular patterning changes in vivo. Specifically, VEGF164 increases the number and total length of <25 μm diameter vessels and increases the numbers of capillaries relative to the microvessels, presumably arterioles, in the 15- to 30-μm diameter range directly upstream from them (Fig. 6). These patterning changes are spatially localized to within a radial distance of 1 mm from the growth factor delivery source, whereas regions of tissue neighboring the microbead are unaffected. These data are consistent with previous results that illustrate the ability of VEGF to induce capillary sprouting (8, 14, 29, 35) and increase tissue vascularity in healthy tissue (12).
Although total vessel length increases are significant throughout day 14, these patterning changes are transient and regress to control values by day 21 of the study (Fig. 6A). We suggest the regression seen at this time point is in part due to the lack of growth factor stimulus present at this later time point, because the detectable diffusion gradient was shown to disappear after the first 3 days. We suggest that because the growth of newly formed microvessels is not accompanied by an increased metabolic demand, their presence is not essential to the function of the tissue. Therefore, the newly formed vessels may be eliminated by the natural regression process (4, 27), leaving only those vessels required for meeting metabolic demands. Consistent with our observations is experimental proof that short-term stimulation by VEGF causes transient remodeling responses (11). We hypothesize that beyond the 21-day time point network, architectures remodel to accommodate the metabolic needs of the tissue, whereas metabolically unnecessary vessels regress without additional subsequent treatments of exogenous VEGF164.
After 4 days, single application of Ang-1* transiently increased the percentage of vessels covered in SMA-positive cells (Fig. 6D). Interestingly, VEGF164 also stimulated a subtle prearterialization response at days 7 and 14 through the recruitment of perivascular cells expressing SMA. This conclusion is supported by the fact that VEGF164 induced significant total length density increases at these time points, but the length of vessels coated by SMA as a percentage of total vessel length did not change (Fig. 6D). Therefore, the total length of SMA-positive cell coverage increased to maintain this percentage near a constant value during these time points. This observation is consistent with previous work that suggests VEGF164 has the ability to protect endothelial cells from undergoing apoptosis (3) and promote vessel maintenance (16, 20). Exogenous treatment with VEGF164 alone has been shown to induce the formation of numerous enlarged vessels in the skin, which later remodel by acquiring a smooth muscle cell coat and function as medium-sized arteries (36). Despite changes in SMA-positive cell coverage, 1:2 order ratio values (which are increased in response to VEGF164 treatment) did not reflect an increase in arteriole relative to capillary growth (Fig. 6C). These data are not contradictory, however, because order ratios reflect branching patterns and are independent of vessel length density changes.
In achieving spatially directed network patterning, we demonstrated the importance of delivering a focal dose of growth factor by subjecting window tissues to a negative control of a single-time point superfusion of VEGF164. Although the concentration (protein/tissue volume) of superfused growth factor was comparable to the average concentration experienced by tissue within the region of focal VEGF164 diffusion, as confirmed by the radioisotope-labeled VEGF164 study, the superfusion delivery method did not generate significant patterning changes. In fact, a superfusion of VEGF164 caused responses similar to those seen when vehicle control was superfused, and neither treatment significantly altered network patterning. Therefore, we suggest that the localized diffusion gradient provided by the alginate microdelivery device is necessary for eliciting significant patterning changes. This conclusion has important implications in tissue engineering and therapies where it would be advantageous to direct vascular remodeling to or within localized tissue regions.
Ang-1 and VEGF are thought to play coordinated roles in vascular development (25, 40) and remodeling (15, 23, 30). It has been suggested that the production of VEGF and the angiopoietins must be spatially and temporally correlated to ensure a normal network architecture remodeling response (28). Transgenic mice overexpressing VEGF and Ang-1 exhibit increased dermal microvessel density compared with mice overexpressing either VEGF or Ang-1 alone. Microvessels in double transgenics were tortuous, capillary-like, and had increased diameters and lengths. However, unlike VEGF-overexpressing vessels, they were not excessively leaky (44). When Ang-1 was administered exogenously in a mouse corneal pocket, it did not induce significant capillary sprouting, but when coadministered with VEGF, Ang-1 significantly increased vascular density and perivascular cell recruitment. Furthermore, perivascular cell recruitment in VEGF-induced neovascularization was dependent on Ang-1; without it, vessels were unable to recruit perivascular cells during the first 6 days (2). These studies suggest Ang-1 functions concomitantly with VEGF to form mature functioning networks.
By sequentially applying VEGF164 and Ang-1*, we induced prolonged patterning changes that were distinct. The transient patterning response, as assessed by functional length density and diameter distribution, that was stimulated by a single dose of VEGF164 on day 1 was maintained to day 21 by delivering a subsequent dose of either VEGF164 or Ang-1* on day 7 (Fig. 8, A and B). Sequential treatments of VEGF164 almost doubled in the number of capillaries relative to the number of arterioles directly upstream (Fig. 8C). However, a secondary treatment with Ang-1* returned to and maintained the 1:2 order ratio to control levels by day 14. This change was accompanied by an increase in SMA-positive vessel length (Fig. 8D). These data suggest that the initial VEGF164 dose induced angiogenesis, and specifically increased total length of capillaries, whereas the secondary dose of Ang-1* maintained the normal vessel hierarchy and was associated with the recruitment of SMA-positive cells.
In a recent study, exogenous delivery of Ang-1* restored normal hierarchical vasculatures in retinas of murine neonates where perivascular mural cells were previously blocked from being recruited by injecting an antagonistic antibody against PDGF-receptor-β (45). Without exogenous Ang-1*, vessels were leaky, and the network pattern hierarchy was abnormal with increased total vascular density and enlarged trunk and capillary vessels. Delivery of Ang-1* did not reinstate perivascular cell recruitment in this model system, suggesting that this growth factor maintains endothelial cell integrity by imparting its effects directly on endothelial cells and not via perivascular cell-endothelial cell interactions. However, this study does not discount the possibility that Ang-1 stabilization of endothelial cell integrity may permit or even promote other downstream growth factor signals that facilitate perivascular cell recruitment. Our data showing that Ang-1* promoted perivascular recruitment and reestablished the 1:2 order ratio when used in conjunction with exogenous VEGF164 support this hypothesis.
In conclusion, previous work has suggested that growth factors function in a coordinated manner during angiogenesis, arterialization, and regression (6, 18, 33, 34), yet how they interact to produce pattern changes at the network level is not well understood. The in vivo approach of this study employs a series of metrics to describe whole network patterning changes in response to exogenous stimulation by spatially and temporally controlled applications of VEGF164, Ang-1*, and combinations of these growth factors. We have provided evidence for a coordinated role of VEGF164 and Ang-1* in generating remodeled networks with native patterning characteristics. In addition to studying the remodeling effects of multisignal combinations of growth factors, we have also demonstrated the ability to perform spatially directed angiogenesis and arterialization, a capability of importance to therapeutic vascularization strategies and tissue engineering.
The authors thank George D. Yancopoulos and Regeneron Pharmaceuticals for donating the Ang-1* growth factor protein, the laboratory of Dr. Joel Linden for supervising the radioisotope-labeling portion of this study, and Dr. Ioamnis Constantinidis of the University of Florida for supplying the alginate beads.
Sources of support for this study include National Heart, Lung, and Blood Institute Grants 5 T32 HL-07284-25 to S. M. Peirce, HL-52309 and HL-65958 to T. C. Skalak, and HL-66307 to R. J. Price.
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 © 2004 by the American Physiological Society


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Fans of the Civic Type R can rest easy in the knowledge that Honda is working on a replacement for the red hot version of the company's small hatchback.
Although the Type R was killed off earlier this year it seems stories of its ultimate demise may be exaggerated.
Appropriately, given the Civic Hatch is manufactured in Britain, there's a Monty Pythonesque tag to the Type R's current status. Like John Cleese's Norwegian Blue parrot, it's apparently not dead; just resting.
According to Honda Europe spokesman John Kingston the lag between the previous and upcoming versions of a performance Civic is due to the imminent roll out of new engines, which will include more fuel efficient petrols and diesels, plus something a bit more special.
"We haven't formally confirmed [a Type R replacement] but it's looking increasingly likely. We will be producing a sporty version of the new Civic," he said.
"The feedback we receive from customers and media is that there's an expectation, there's a hope, there's a desire that Honda is going to bring back the Type R so that's the direction we want to head."
The previous Type R expired when the latest Euro V emissions regulations came into force early this year; its high-revving 148kW 2.0-litre naturally aspirated (non turbo) engine were unable to meet the stricter emissions.
The new Civic Hatch gets updated versions of the existing (and milder) 1.8-litre engine, being used a s a stop-gap measure until the range of so-called Earth Dreams drivetrains is rolled-out progressively from late this year.
"The development program around the new Civic is to look to add to the Civic range throughout its lifecycle so you'll see the introduction of the 1.6-litre diesel at the end of this year" Mr Kingston said.
"It's then likely we'll see the introduction at some stage of a more sporty Civic and then we'll look to potentially add new variants as well."
Neither Mr Kingston nor Civic project leader Takehiko Masuda could confirm Honda was looking at a lower revving turbocharged engine for the Type R replacement as a way of solving both the performance and emissions dilemmas. Many compact hot hatches it would ultimately compete with – from the Subaru WRX and Volkswagen Golf GTI to the Drive Car of the Year category-winning Renault Megane RS250 – use turbocharged engines.
Current information released on the petrol four-cylinder Earth Dreams power units indicates they will be non-turbo engines ranging from 1.4 to 2.4 litres in capacity, and using a more efficient Atkinson Cycle combustion process (used in some hybrids) to control fuel consumption and emissions.
One option could be a hybrid drivetrain; the Honda CR-Z is proof the company considers hybrid a viable proposition for performance models.
However, Mr Masuda did not rule out turbocharging as a way of delivering more performance.
"We are looking at every at every technology and method to achieve that goal," he said.
Mr Kingston said Honda had deliberately delayed its product replacement program, citing as factors such as the global economic crisis and last year's Japanese earthquake, which damaged the company's Tochigi research and development centre.
"We acknowledge that the last two or three years have been difficult for us. We took some decisions to slow our product replacement, we're coming out of that period now and we'll respond with new products and new engines," he said.
When the Type R replacement does hit the streets, it'll be an almost certain starter for Oz according to Honda Australia director Stephen Collins.
"If there is one, we'll be sticking our hand up for it," he said.
Follow Drive.com.au on Twitter @Drivecomau

subset: commoncrawl


text: Hey guys just picked up my cobra the other day. The owner told me that the cd player doesnt work in it so it has to be replaced. My question is if I throw an aftermarket player in does it still use all 8 speakers? Or do I need a special head unit for it?


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Chicago - A local Illinois hybrid of the Bilderberg Grope is courting Chicago Mayor Rahm Emanuel for his Vice Presidential aspirations as the Dems' official White House 2016 running partner.
A rumored $100 million war chest is sitting pretty ready to roll former city slicker Rahm regardless of the outcome of the presumed Hillary Clinton/Joe Biden presidential nomination face-off.
"Rahm's our best guess for Veep 2016," a spokesperson for Dem Bundlers Anonymous said in reply to reports that a wall of hedge fund cash will back President Obama's former Chief of Staff.
"Hell, most of those vulture funds were originally backing Amazonbotch's $19 billion Chevron lawsuit," the DBA source added, "before snatching their money back and turning tail."
53 year-old ex-Bill Clinton aide and sometime investment banker Rahm Emanuel is now these hedge funds' golden boy having impressed with some panache his unique campaigning skills honed under the tutelage of the Comeback Kid.
This included sending a dead monkfish in a box to a bewildere pollster who'd hinted in public about 'something fishy' going on with Rahm.
Media reports at that time likened the gesture to the severed horse's head scene at the start of the movie The Godfather but the publicity never harmed a single hair from Rahm Emanuel's head.
Might even get him the conspiracists' vote in 2016.


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Our active team of librarian bloggers deliver fresh content every week. Find new arrivals, information about upcoming library programs, as well as library news and updates.
By Staci T.
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Here's a sampling of our new nonfiction book, audiobooks and DVD arrivals. Click an image to view availability or place a hold.
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By Staci T.
APL is hosting an Indiana Blood Center blood drive on Thursday, August 22. The Bloodmobile will be in the APL parking lot from 9:30-11:30am. Schedule your donation appointment now!
By Danielle A.


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Functional Assessment Screening Tablets - Patient Reported Measures
This study has been completed.
University of Pittsburgh
ClinicalTrials.gov Identifier:
First Posted: March 3, 2010
Last Update Posted: January 15, 2016
Information provided by (Responsible Party):
Rachel Hess, University of Pittsburgh
|Tracking Information
|First Submitted Date ICMJE
|February 27, 2010
|First Posted Date ICMJE
|March 3, 2010
|Last Update Posted Date
|January 15, 2016
|Start Date ICMJE
|March 2010
|Primary Completion Date
|October 2010 (Final data collection date for primary outcome measure)
|Current Primary Outcome Measures ICMJE
|Doctor Patient Communication [ Time Frame: The doctor and patient pair will complete a one time survey within two weeks of the doctor patient encounter. Audiotaping of the encounter will occur once at the time of encounter. ]
the doctor patient and encounter will be queried for communication regarding the patient reported measures
|Original Primary Outcome Measures ICMJE
|Same as current
|Change History
|Complete list of historical versions of study NCT01080183 on ClinicalTrials.gov Archive Site
|Current Secondary Outcome Measures ICMJE
|Not Provided
|Original Secondary Outcome Measures ICMJE
|Not Provided
|Current Other Outcome Measures ICMJE
|Not Provided
|Original Other Outcome Measures ICMJE
|Not Provided
|Descriptive Information
|Brief Title ICMJE
|Functional Assessment Screening Tablets - Patient Reported Measures
|Official Title ICMJE
|Functional Assessment Screening Tablets - Patient Reported Measures Translating Research Into Practice Pilot
|Brief Summary
|The team's ultimate goal is to improve health care quality and effectiveness and support patient-centered care. In this project, we, the investigators, will test whether providing patients with guideline based-recommendations regarding tobacco use, physical activity and mental and physical health-related quality of life (patient reported measures: PRMs) using health information technology increases doctor-patient discussions regarding these topics and results in improvements in PRMs. We will randomize physicians in a single general internal medicine practice. Patients seeing intervention physicians will receive guideline-based recommendations regarding PRMs, those seeing control physicians will not. We will compare differences in doctor-patient discussions regarding PRMs between the intervention and control groups. The successful completion of this project will provide evidence the effectiveness of involving patients in their care through the use guideline-based feedback.
|Detailed Description
|Not Provided
|Study Type ICMJE
|Study Phase
|Not Provided
|Study Design ICMJE
|Allocation: Randomized
Intervention Model: Parallel Assignment
Masking: Single (Outcomes Assessor)
Primary Purpose: Health Services Research
|Condition ICMJE
|Intervention ICMJE
|Other: Patient feedback form
patients receive a report that feeds back their patient reported measures to them prior to their clinical visit
|Study Arms
|Publications *
|Not Provided
* Includes publications given by the data provider as well as publications identified by ClinicalTrials.gov Identifier (NCT Number) in Medline.
|Recruitment Information
|Recruitment Status ICMJE
|Enrollment ICMJE
|Completion Date
|October 2010
|Primary Completion Date
|October 2010 (Final data collection date for primary outcome measure)
|Eligibility Criteria ICMJE
Inclusion Criteria:
Exclusion Criteria:
|18 Years and older (Adult, Senior)
|Accepts Healthy Volunteers
|Contacts ICMJE
|Contact information is only displayed when the study is recruiting subjects
|Listed Location Countries ICMJE
|United States
|Removed Location Countries
|Administrative Information
|NCT Number ICMJE
|Other Study ID Numbers ICMJE
|Has Data Monitoring Committee
|U.S. FDA-regulated Product
|Not Provided
|IPD Sharing Statement
|Not Provided
|Responsible Party
|Rachel Hess, University of Pittsburgh
|Study Sponsor ICMJE
|University of Pittsburgh
|Collaborators ICMJE
|Investigators ICMJE
|PRS Account
|University of Pittsburgh
|Verification Date
|January 2016
ICMJE Data element required by the International Committee of Medical Journal Editors and the World Health Organization ICTRP


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What a difference new (and hopefully) stable ownership makes.
Only days after getting a reprieve from Glendale City Council that will keep them in Arizona for the next five years, the Phoenix Coyotes jumped into the NHL free-agent market, landing the highest-scoring player available: centre Mike Ribeiro.
Ribeiro played for Coyotes head coach Dave Tippett in Dallas from 2006-09, which was a factor in his decision, but so were the dollars general manager Don Maloney could spend. In this case, US$22-million over four years, plus the stability of knowing the team was staying and playing in Arizona.
"Normally, we're hitting the phones begging: 'Will you please come and take a leap of faith?'" Maloney said with a rueful laugh Friday, "And that just gets old after a while. This was such a totally different experience.
"It just made our life 100-per-cent easier. Now, it was just a matter of 'who do we want?' and 'what can we afford?' There wasn't any discussion of where we're going to play. So that was a good thing."
The Coyotes were like a lot of Western Conference teams at the end of the first day of free agency, shuffling pieces in and out, and hoping they got a little better overall.
They lost backup goaltender Jason Labarbera to the Edmonton Oilers, but replaced him with Thomas Greiss, the former San Jose Sharks backup. They also lost checking centre Boyd Gordon to the Oilers, but replaced him with Ribeiro, a far-more dynamic player, who – according to Maloney – gives them the play making centre they've been seeking for the past six years.
The Winnipeg Jets got in on the action, acquiring right-winger Devin Setoguchi from the Minnesota Wild for a second-round pick in the 2014 draft.
The Oilers were the most active Canadian team in the West, adding defenceman Andrew Ference (Boston Bruins) on a four-year, $13-million contract. Ference is from Sherwood Park, Alta., and tweeted a picture of himself as a young child, wearing an Oilers sweater, just before the announcement of his signing became official.
He and his wife have family in the area, so there was a lot of compelling off-ice reasons for Ference to sign there. But the overriding decision was the Oilers' potential – a team that has invested four seasons into a scorched-earth rebuilding program and has missed the playoffs for the last seven.
In Ference, the Oilers get a relatively undersized defenceman who nevertheless has played in three Stanley Cup finals since 2004 (two with the Bruins, one with the Calgary Flames). He became expendable in Boston largely for salary cap reasons, but also because the Bruins had a handful of NHL-ready defencemen in their player pipeline.
Gordon will replace Eric Belanger on the Oilers roster as a penalty-killing and faceoff specialist, while Labarbera will be the backup to No. 1 goalie Devin Dubnyk. The Oilers also completed a trade that sent captain Shawn Horcoff to the Dallas Stars for defenceman Philip Larsen and a 2016 seventh-round draft choice.
The fact the Oilers could trade Horcoff rather than issue him a compliance buyout had a lot to do with the structure of his contract, which was heavily front-loaded. While it carries a salary-cap charge of $5.5-million, the actual dollar amounts he will receive for the next two years are $4-million and $3-million respectively.
For a team such as Dallas, which won't be spending to the cap any time soon, the extra cap charge doesn't matter at all. In fact, it was just two seasons ago, when the Stars had to add players in order just to spend to the salary cap floor.
The additions will help Edmonton GM Craig MacTavish reshape his team, but do not represent the sort of "bold" moves he promised when he took over from Steve Tambellini.
All-sports cable TV channel TSN calculated $360-million had been committed to 52 players through the supper hour, which was about $140-million ahead of last year's pace.
A year ago, however, the biggest dominoes, Zach Parise and Ryan Suter (both to the Wild), didn't fall right away. They eventually added almost $200-million to the overall spending tally, which never seems to abate, no matter how many player lockouts they go through.
Ference and others were able to make their decisions more quickly this time around because of a two-day negotiating window written into the new collective agreement, which permitted prospective free agents to speak to potential suitors before the signing window officially opened.
The only thing that didn't change: Players continued to reap huge financial rewards and Maloney, for one, said he didn't expect to see that change any time soon.
"The cap for next season is artificially in place," the GM said. "After that, when we did our projections of where it's going, nobody seems to think it's going down. When you start adding the really great job the NHL has done in marketing the sport – adding in the outdoor games and the Winter Classics and all the different revenue-generating possibilities – who knows where the cap goes, which is directly related to revenues?
"If I've learned anything in the last two days, if you have anybody you like, you better sign them sooner rather than later – and don't let them get close to free agency."


text:
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Dragon Age 2 - David Gaider Interview @ Geek and Sundry
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April 30th, 2012, 07:16
I watched the whole thing. (yes, I was bored and had nothing better to do.) Its basically a fan girl of Bioware asking silly and obvious questions to David Gaider and then making obvious reactions to his answers. It can be safely skipped, nothing interesting. Unless you are a "super" fan boy/girl of the Dragon Age universe or something like that I suppose. (They kept mentioning characters by name and talked as if they are intimately familiar with them, whereas even though I played the first Dragon Age game, I found the characters to be very forgettable.)
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text: Lancaster is the county town of Lancashire, and the seat of the Duchy of Lancaster. Settled since Roman times, its center boasts a number of beautiful buildings, notably the Ashton Memorial, Lancaster Castle and St Mary's Church. The city has a thriving cultural life, with art galleries, music venues, and museums in abundance. A large firework festival is held around Bonfire Night every November.Hide


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Keep your elbow pinned to your hip, your forearm perpendicular to your body, and your palm facing the sky. Then push hard with your back leg just like you would when you're swinging for a line drive, which makes you fire your hips through, which makes you fire your forearm through, which makes you snap your wrist and eject the disc.
Your body becomes a whole series of levers, just be sure to have them all back, cocked, and loaded to get triggered by the previous lever. If you do it in order you build up a ton of force with minimal movement. If you don't do it in order the energy isn't efficiently transfered through your body. Ie: If you fire your arm through before your hips then the force from your hips and legs doesn't get into the throw.
In the end it should feel like you're pushing hard with your legs and hips and your forearm and wrist just fire through on their own because they don't have a choice. So you're forearm basically just becomes a lever that gets rocketed through by your hips. That Sarah Hokom video is a perfect example. Watch her thighs flex and propel everything through.
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text:
I found a large starfish in the ocean about 4 days ago. I left it out in the sun for 4 days trying to dry it out. It still has the smell and I was wondering what I should do now to preserve it and get rid of the smell?
Return to Reptile Taxidermy Category Menu
You should have sprinkled salt over it as soon as you brought it home . This will help it dry out but you better get used to the smell , it never goes away . I think it smells pretty good myself . It brings back memories of the beach when I was a kid . Now I live at the beach and never get to go there , too much work . Rick


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The past few months have seen the launch of two hot San Francisco startups with mobile photo-sharing apps — Instagram and Picplz. Today, Picplz announced that it's catching up with the competition on the social networking front, by adding support for photo-sharing site Flickr as well as blogging services Tumblr and Posterous.
I'm not sure if it's fair to paint the two startups as being in a head-to-head rivalry, but both apps seem to be built around the same basic idea: Users take a photo, add a filter, add location information, then share the photo within the app and other social sites. The startups even share an investor in Andreessen Horowitz, though partner Marc Andreessen has said that he will take a more passive role in Instagram.
Picplz has the advantage of working on both Android and iPhone, while Instagram seemed to be ahead in support for social networks. Now the tables have turned, with both apps allowing you to post to Twitter, Flickr, Facebook, Tumblr, and Foursquare, and only Picplz supporting Posterous.
At this point, the exact laundry list of services probably isn't that significant, but I think it's important for these apps to have a wide social reach — the selling point of an app like Picplz is the ability to share your photos anywhere, not just within the Picplz community.
Picplz was co-founded by Dalton Caldwell, who also founded music-sharing service Imeem. The company recently raised a $5 million first round.


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By: Beatriz Craven, Guest Blogger
All Images Courtesy Beatriz Craven
"It's just stuff, it's just stuff, it's just stuff" I repeated to myself as I handed over bag after bag of my belongings to Goodwill. Some included Gucci sunglasses from a few seasons ago. Others were filled with pretty linens, handbags, and little finds that had managed to make their way into my home and life. My husband and I were relocating from the Houston suburbs to the city. As a result, we were downsizing. Significantly. While we could probably just get a storage unit for our extra belongings, I decided to take on the challenge of editing our things down to the essentials. It seems more dramatic that way, and I kind of like it: a real test to my sense of materialism.
I was born in South America and came to the United States as a privileged child. I inherited my mother's golden standard for brand labels and good quality products as a "must." I'm sure some of this has to do with social class. The rest I believe is due to trusting brand names in a third world country. There always seemed to be a sense of comfort that came with knowing you had the real deal. Even now, I never buy store brand. God forbid my Advil look alike is actually nothing more than a sugar pill in a cheap box from Lord knows where (I'm going to go ahead and blame this delusional thinking on my parents). We are such a money and possession driven culture. I realize how easy it can be to get caught up in nice things (or wanting nice things) and I make it an honest effort to always remain grounded. Hence the Goodwill challenge. So when my husband asked me to go on a camping trip with him for two weeks I say let's do it. I love being in nature and I'm sure I can handle giving up our daily comforts for the camping life for a while.
Well people, let me tell you something: two weeks of camping is a long time, enough to break a woman. Or well, to break me I guess. Don't get me wrong, I was really enjoying the incredible nature, but in those first few days I had a really hard time surrendering myself to camp life. I could feel myself dragging my feet as we set-up the tent. Again. If I could curse out every last one of those little bugs, I would have. I became incredibly annoyed by the male capacity to whip it out and pee when I had to get friendly with the bushes and the trees in a way that I really didn't want to. And on top of all that, I had to take off my nail polish. The horror! I hadn't gone with bare nails since I was pre-pubescent. True story. It was finally time for a come to Jesus meeting.
I got over myself. I'm pretty sure it was our stop to shower at a place that required quarters for hot water that finally pushed me to a place of no shame. I would imagine it's kind of like how a woman feels after giving birth and all those people see your vagay in all her glory. After that you kind of don't give a damn anymore. Having reached my surrender, I went from enjoying the trip to truly loving the trip. There are some utterly fantastic places to visit in this country, and I was glad we got to experience it in such a cool, primitive way. It wasn't until our drive home that I realized the extent of my outdoors adaptation. We stopped at a Jimmy John's for one of their ridiculously delicious sandwiches and I used their facilities. I had a legitimate reaction of awe upon entering their restroom. "This place is BEAUTIFUL!" I gasped. A good indication that it was probably time to rejoin society.
First world problems. It's so easy to get caught up in the luxuries. We don't have to get crazy, but it's so important to look past the superficialities of the world and get to what's real. That's where we find the good stuff. Plus, it can be kind of fun to rattle our own cages every once in a while and see what happens. You never know, you might surprise yourself with the joys that come with learning to live without.
About Beatriz: Beatriz Craven is desperately close to fulfilling her dream of becoming a psychologist with the completion of her PhD. She is an avid life enthusiast, loving wife, and movie fanatic.


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Anne in the Stands: Brewers fever
Team's successful campaign bringing about more fans
By /
With the Brewers monopolizing headlines not only at home but around the country, I've noticed these impressive effects:
Everywhere I go -- grocery store, gas station, corn stand or the airport -- people of all ages are wearing Brewers hats and shirts. I love that.
My buddies, who don't know what a 6-4-3 double play is and can't name another team in our division besides the Cubs, are suddenly eager to talk baseball. I love that, too.
My stadium is crowded with cheering fans. I love that even more.
Hordes of people like to park in my favorite section of the stadium lots. I don't love that. As I hike from my parking place in no-man's land past jillions of cars and squads of tailgaters, a couple of thoughts occur. First, will I remember where I parked? Next, I acknowledge that sellout crowds pay for talent on the field.
Oh Pain, oh agony, oh what you do to me: A confession from the depths of a baseball funk
This morning my computer announced I was protected against 182,701 threats. I suppose I should feel good about that, but what I really need is a magic potion for pitchers who can't get the ball across the plate and for batters who can't get a hit with men on base. It wouldn't hurt to get my team immunized against the haunting presence of the Chicago Cubs.
You could call my affliction "baseball funk." When the Brewers win, birds sing. The grass is green on my side of the fence. Life is good. I even like how I look in the mirror. But when they lose, the sky's gray. The flowers in the back yard lose their colors. I can't hit a tennis ball, I can't spell and everything I put on makes me look fat. The gloom lingers until Prince Fielder hits another homer, T-Bow aces three batters in the eighth and Francisco Cordero closes out the ninth with just eleven pitches.
As I agonize over every pitch and every at-bat, I wonder if I'm crazy to love my team so much that my husband prefers to stay out of my way for awhile after a loss. He calls it "serving time in the penalty box," to mix in a winter sport metaphor. I'm lucky he's so tolerant.
The other day, Mark, the man who sits in front of me at Miller Park, counseled me not to panic over losing streaks or pitching meltdowns. "It's a long season," he said. "Just hang in there."
I'm taking his advice by enjoying the summer weather and all the other aspects of my life, and by remembering the many great games we've seen this season. I'll hang in there to cheer on Prince, J.J., Johnny, Derrick, Corey, Ryan, Geoff, Damian, Ben, Cappy, Dave, Yovani, Tony, Craig, Billy, Kevin and the rest of the team.
No matter what happens.
This story was not subject to the approval of Major League Baseball or its clubs.


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I think much like The Situation that Bristol could potentially take out better dancers not due to her dancing but her political affiliation. It will be interesting to see if her scores start to suddenly rise beyond her ability.
I think much like The Situation that Bristol could potentially take out better dancers not due to her dancing but her political affiliation. It will be interesting to see if her scores start to suddenly rise beyond her ability.
The sad thing is, they'll have to take Bristol and Mike's scores higher at some point. They can't keep giving performances as bad as last night's were, but continue to praise them and yet still give them the exact same scores they keep getting. Even with horrible performances, eventually it'll grow old, but if they don't improve, it's going to be hard to keep praising them strictly for ratings purposes and giving them 8s and 9s. Mike didn't deserve 7s last night, or even a 6. He should have scored below Margaret.Originally Posted by ChelsieFan;4077691;
I don't know. Their DialIdol scores were much higher than the next lowest pair, for whatever that's actually worth. I know that measures the phone line busy signal and not online or text voting, but if all is relative, her mom's fan base is really going to bat for her. In fact, I believe she has lead in scoring on DialIdol the last 2 weeks also. The discrepancy in that scoring is a bit disconcerting.Originally Posted by karalott;4077744;
Last edited by MFWalkoff; 10-05-2010 at 10:03 PM.
Notice that Tom never indicated that she was in the actual bottom 2. She was in jeopardy because she got the 2nd lowest judges' score.Originally Posted by TiaMaria62;4077753;
He was very careful with his wording...."but did the viewers choose to save you?"
She was at the top of the heap on that list which meant she got a lot of calls. It wasn't off that much!
Something that I have noticed is that they are lining up the contestants solely by the judges' scores. Since we don't get an accounting firm's verification, we have no idea whether the person who goes was voted off by the viewers or not.
Good point. I will start voting when they can show proof of where the votes go. I don't take any vote count serious (with any of these shows) without proof. The network can say anything, without proof I don't believe them, show me the actual vote number.Something that I have noticed is that they are lining up the contestants solely by the judges' scores. Since we don't get an accounting firm's verification, we have no idea whether the person who goes was voted off by the viewers or not.As far as any comparison between Kelly and Bristol, there is a big difference, Bristol is a pleasant unassuming lady. Kelly cried all through the show "poor little me" type person. Kelly had a very compassionate pro partner Louis who everyone likes. Louis tries to make it about his celebrity partner. Bristol has Mark, who no one likes. To Mark it's all about him such a showoff ham. With a different partner (like Louis or Tony) perhaps Bristol would do better. She definitely should have had a taller partner to match her height. All in all, I'd rather watch Bristol trying hard in her quiet way than watch Kelly crying the poor little me jag for the sympathy card. The judges were very conscious of the Osbourne/Simon Cowell friend name when judging little Kelly. Hopefully, Bristol will get the same respect without being in the final three (like judges pushed Kelly to). She doesn't deserve to be put through over better dancers like Kelly was. No celebrity on this show deserved the disrespect Bruno showed Boden. Yes, judge them harshly but don't rip them to shreds with a public flogging. It's not entertaining to me. They all deserve some manner of respect for trying.I doubt that she can match Kelly's transformation, but she apparently has had some past athletic experience, and seems to have started the show in a little better physical shape than Kelly, so I suppose it is possible. On the other hand, I don't think Bristol has as much determination, her partner is less helpful, and the competition this season is stronger.
Last edited by Arielflies; 10-06-2010 at 03:07 PM.
Kelly may have been more emotional or more of a drama queen than Bristol, but she also had personality, charisma and a natural enthusiasm that came across in her dancing. Bristol, from what I have seen so far, has none of that. Not surprising, since Kelly was a performer, from a show biz family. Bristol, apart from her famous mother, is an unexceptional and somewhat dull young woman, and that is what comes across in her dancing to me. Yes, she has done a good job learning the steps and I'm sure she's trying hard, but I suspect that if you chose a bunch of random ordinary people off the street and taught them each a dance routine, you'd end up with a lot of performances like Bristol's. Very nice, but I don't need to watch it. At this rate, if she (or Mike) make it into the top 6 when they do the team dances, it will really be a travesty and totally unfair to the other dancers. Feh.Originally Posted by Oceansands;4078208;
Who the heck is voting for Bristol and why is she still there. She should have gone home last night, not Cho. I wonder if Bristol's nutty and crazy mother will be there next week.
I don't think that I can blame Bristol's singular lack of charm on her background not being show biz. I have seen many "average" people over the years on television (man on the street, being interviewed on talk shows for events in their life, etc.), and that includes teenagers and children, who were chockful of charm, wit and charisma. She is just dull. She is a dull dancer, she has a dull personality, and she appears to be intellectually dull based on what she has to say to the camera.Originally Posted by ironcat;4078218;
And so it begins, the end of times..
I strongly disagree that Margaret was even nearly as good as Bristol. Though this story telling thing requiring emotion wasn't her forte, still her footwork, her arms and the grace with which she moves were head and shoulders above Margaret who just looks awkward.
I don't think people should vote for her because of her mother, but neither do I think she should be penalized for the same reason.
(And yes, I agree she's been overscored - almost everyone has been.)
True, but every opportunity she's had over the past two years has been because of who her mother is. Doing this show might actually provide her the opprtunity to break out and get further offers because she's Bristol Palin from DWTS, not just because she's Sarah Palin's daughter.Originally Posted by ChelsieFan;4077112;
I thought her dance was dull this week, but that's Mark's fault not hers. I thought she did ok with the little bit of acting at the beginning of the dance. She was a little wooden, but at least she attempted to act her part rather than just stand there.
"That's Numberwang!"


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Written on: 03/08/2008 by wendywendy (3 reviews written)
Good Points
Bad Points
All other features on the phone.
General Comments
I think these Philips ID937 phones are hopeless! I have had Philips phones for years and they have been fine (the Karla Vox was the best of all) but these ones are useless. The ring tones are poor and even when they are turned up to full volume they are still hard to hear. You can't alter the number of rings to more than the settings therefore i am forever missing answering calls before they go onto the answer machine. As for the volume control, it is terrible, you can hardly hear the person on the other end despite turning the volume up to 5 (and no, I'm not old and deaf). I have only had these phones for 6 months but I will be changing them soon.
" Don't trust this service " Read More
" ICreation I650 DECT cordless phone - very good but limited " Read More
Written by IanLHarris
" Chinese Junk " Read More
Written by wilko50
" Best set of dect phones " Read More
Written by rodbradford
" Cheap but unclear " Read More
Written by Greggory


text:
China's Internet Triple Speak
From The Diplomat:
On the possible reasons for the Chinese decision, [Rebecca MacKinnon] says it appears that the pragmatists within the Chinese government won out. It's a reasonable observation—as she notes, refusing the license would have meant shutting Google out of China completely, thus sending an extremely negative message to the international business community.
If this is the case, it comes at a good time for China, following criticism from the likes of GE Chief Executive Jeffrey Immelt,who earlier this month took a rare pubic swipe at the Chinese government by stating that the country was becoming increasingly protectionist. According to the FT, Immelt pondered out loud in his a speech whether, 'in the end they (China) want any of us to win, or any of us to be successful.'
On the issue of the Internet, at least, Chinese government officials won't hear of any suggestion that they're anything but open. Indeed, last month the government released a white paper on its Internet policy in which it claimed that: 'Chinese citizens fully enjoy freedom of speech on the Internet.'
Read MacKinnon's relevant blog post here.


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BENTON, Ark. (KTHV) – The 2014-2015 school year is just weeks away, meaning police will be increasing patrols in school zones.
The Benton police say for the whole month of August they will be cracking down on speeding and distracted driving.
"We are going to be out in full force looking for any kind of violations in school zones and we'll have officers as we have in the past. Officers will be randomly assigned to get out the of their vehicles throughout the day and to walk through the schools and to let the teachers and students know that we're here if we need them," said a Benton police officer.
The Benton police say you can find information about the initiative on their Facebook. It will continue up until the Labor Day holiday.


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KANSAS CITY, Mo. — When the American League schedule first came out about 10 months ago, this week's Oakland-Pittsburgh matchup was lost in the crowd, just another interleague series among dozens of them.
In the wake of the A's 10-4 win over the Kansas City Royals on Sunday, it's a showdown between two of just four big league teams that have 52 or more wins and, some say, a precursor to a World Series showdown.
That last bit is something of a stretch, given that there is almost half a season still to play and baseball hasn't gotten to the All-Star break yet, much less to a playoffs push. Still it's intriguing to see the Pirates, who haven't been to the postseason since 1992, and the A's, who have made it out of the first round of the playoffs just once since '92, squaring off.
"Pittsburgh is playing very, very good," A's manager Bob Melvin said. "Just look at their pitching numbers.
The Pirates' team ERA of 3.15 is the best in the major leagues this year, and the three starters Oakland will see — left-hander Jeff Locke, right-hander Gerrit Cole and lefty Francisco Liriano — are a combined 20-5. Small wonder Pittsburgh has 53 wins, a total surpassed only by Boston.
The A's team the Pirates will host has the second-best ERA in the American League at 3.72 and a team that is starting to put the pieces together offensively. That starts with right fielder Josh Reddick, who matched his career high with four RBIs Sunday with a two-run double and a two-run homer.
Reddick doesn't have anything close to great numbers, but he has great potential as his play of the past two weeks suggests. He has reached base safely in 16 consecutive games and has raised his batting average about 20 points to .222.
"It's not so much a confidence issue," Reddick said. "I've always had that. It's been about being able to recognize pitches."
That ability was tested from mid-April to mid-June as Reddick tried to play through a wrist injury, went on the disabled list, then tried to come back from the injury too quickly.
"It was stupid of me to try and play through it," Reddick said. "I couldn't pull the ball at all."
He's just getting back to the point where he was last year when he was as dangerous a hitter as existed on the A's roster. He hit only .242 last season, but he had 32 homers and 85 RBIs and was a rock in the middle of the lineup. With Reddick struggling, others have had to step up.
Josh Donaldson, the not-quite-All-Star third baseman, is the obvious one with his .319 average, 15 homers and 57 RBIs, but there are other less obvious ones. Jed Lowrie, who had three hits, including a solo homer Sunday, has a .301 average and has batted either third or fourth 44 times. And Eric Sogard has lent some offensive impact at second base. With two hits, including his first homer in almost 15 months, he's up to .261 for the season.
"Guys have picked it up," Reddick said. "J.D. has picked everyone up. This is a team effort, not two or three guys."
Sogard, who homered for the first time in his big league career against a left-handed pitcher, may have had as important a hit as the A's had Sunday. The Royals had closed to within four runs and were surging, but all that ended with Sogard's seventh-inning homer.
Jesse Chavez wound up pitching the final four innings without allowing a run, and that closed out Kansas City and set the stage for a major showdown in Pittsburgh, of all places.
Monday's game
A's (Bartolo Colon 11-3) at Pittsburgh (Jeff Locke 8-1), 4:05 p.m. CSNCA


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Mobile Bay Turns Tables in LA Classic
Mobile Bay, second to The Pickett Factor in the $150,000 Louisiana Champions Day Classic Stakes one year ago, turned the tables on that rival and four others in the 1 1/8-mile race for state breds Dec. 10.
Mobile Bay, second to The Pickett Factor in the $150,000 Louisiana Champions Day Classic Stakes one year ago, turned the tables on that rival and four others in the 1 1/8-mile race for state breds Dec. 10.
The top two choices in the Dec. 12 Louisiana Champions Day Classic Stakes at Fair Grounds hooked up in early stretch, but it didn't last long as The Pickett Factor put away Mobile Bay and drew off to win.
Fair Grounds Race Course & Slots will kick off its 143rd season of Thoroughbred racing on Friday, November 21.
Populist Politics could emerge as the new state-bred ruler at Fair Grounds Dec. 8 when he takes on 10 other runners in the $150,000 Louisiana Champions Day Classic, main event of eight restricted stakes in New Orleans.
The current king and queen of Louisiana racing, Star Guitar and Superior Storm, are set to go for their record fourth crown each in the annual Louisiana Champions Day at Fair Grounds Dec. 11.
The Louisiana Champions Day program at Fair Grounds Race Course & Slots Dec. 12 was cancelled after the eighth race due to hazardous weather, which included torrential rain and heavy winds.
Brittlyn Stable's Star Guitar is seeking to become the third Thoroughbred in history to win three times on a Louisiana Champions Day program in New Orleans when he goes forward in the Louisiana Champions Day Classic.
One major upset and the matching of two career records headlined the 18th edition of Louisiana Champions Day Dec. 13 at Fair Grounds.
Nine horses who have won during previous Louisiana Champions Day programs return to Fair Grounds in an attempt to repeat as the track plays host to the event's 18th renewal Dec. 13.
Heiligbrodt Racing Stable's 4-year-old filly Ahead of Her Time, who was named Louisiana's 2007 champion 3-year-old filly, recently fractured a sesamoid during a morning training session and was euthanized.
Short prices warmed the hearts of favorite players as Louisiana Champions Day returned to its home base Saturday at Fair Grounds Race Course. The featured $150,000 Classic went to even-money choice Costa Rising by a nose.
Fair Grounds Race Course used the spotlight on Louisiana-breds on Louisiana Champions Day to honor the recently retired Happy Ticket, who became the highest earning state-bred in history last month.
Charles Castille Jr.'s Costa Rising, coming off a win in Delta Downs' Gold Cup last month, faces 2005 winner Badtotheboneandrew and nine others in the $150,000 Louisiana Champions Day Classic Saturday at Fair Grounds Race Course.
Fair Grounds Race Course is now accepting stall applications for the 2006-07 racing season, its first back in New Orleans since Hurricane Katrina.
Charles A. Castille Jr.'s Costa Rising won his sixth consecutive race when he captured the $100,000 Louisiana Breeders' Derby for 3-year-olds Saturday by two lengths to highlight Louisiana Champions Day at Louisiana Downs.
Records were matched, streaks continued, and favorites ruled the 15th annual Louisiana Champions Day Saturday at the Fair Grounds meet at Harrah's Louisiana Downs. Badtotheboneandrew, at odds of 4-1, posted the day's biggest payoff in the $150,000 Classic.
Even-money favorite Spritely Walker enjoyed a ground-saving trip to register a three-quarter of a length victory in Saturday's $150,000 Louisiana Champions Day Classic at the Fair Grounds.
Louisiana's finest will square off Saturday to vie for divisional supremacy in the 13th annual Louisiana Champions Day at Fair Grounds.
Many of the likely top contenders in Saturday's 13th annual Louisiana Champions Day are getting in their final preparations.
The telecast of the 2001 Louisiana Champions Day has won the Eclipse Award for local television, it was announced today. The show was shown on Fox Sports Net Southwest.
Two-year-old Meteor Impact makes the biggest noise on Louisiana Champions Day.
Fair Grounds Race Course in New Orleans presents Louisiana Champions Day Saturday, a series of 10 races worth a total of $1 million for horses bred in Louisiana.
Louisiana's favorite daughter, Hallowed Dreams, faces a field of 10 boys and one other filly in the $100,000 Louisiana Champions Day Sprint Saturday. Winner of the A.B. Leggio Memorial Handicap just one week ago, Hallowed Dreams will carry 120 pounds, including regular rider Sylvester Carmouche.
Louisiana Champions Day is when the little man can stand at center stage, and they don't get much smaller than Peter Orlando. Right On the Mark, a 6-year-old gelding owned and trained by Orlando, gave the 59-year-old his first stakes win in 40 years of racing in the Champions Day Classic Dec. 9. "I'm just a small guy," Orlando explained. "It's just me by myself and four horses. I breed 'em and race 'em. That's all I do."


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Japanese Original Woodblock Print
- Price: $300.25
- Item Category: Antiquities
- Type: Item for Sale
- Date Posted: Aug 03, 2010
- Seller Type: -
I have a Japanese original woodblock print depicting "Princess TOKIHIME, a daughter of a warrior, TOKIMASA ONDA. Artisit: SHOSHIN Circa 1950 This was appraised by FUJI MJRAKAMI, A.S.A. , Oriental Fine Art and Antique Appraiser , Born In Tokyo Japan, Graduated the Chuo University in Tokyo receiving Bachelor of Arts degree in American and English Literature; studies Japanese Classical Literature for two years at the Waseda University in Tokyo. Member of Appraisers Association of America since 1978 and Senior Member of American Society of Appraisers. I can get a better picture of this I believe as my flash reflected on the glass covering the framed item.


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Amazon is offering 5 free holiday songs every 5 days in December, for a total of 25 free songs. You can pick up the current 5 songs now. Not all of the songs will be appropriate for children (sometimes one with questionable lyrics comes up, so do listen to them first).
The current five free holiday songs range from Brian McKnight to Twisted Sister. I kid you not.
We're keeping a list of holiday deals & freebies updated here…..


text: Just wondering if anyone has experienced this? I have a Beseler 23CII and had to replace a bulb yesterday that was about one year old, and now today I have to replace the replacement. The bulbs are GE PH/140 75W. Did I buy bum bulbs or is there something with the enlarger that I need to check? Thanks for any help! Blessings, K


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Twins to skip struggling Liriano's next start
Twins to skip struggling Liriano's next start
Minnesota has an off-day on Thursday, which allows the Twins to go with a four-man rotation before Liriano makes his next start May 1 in Anaheim vs. the Angels. Liriano, who allowed five runs in five innings on Sunday against the Rays, has an 11.02 ERA with 12 strikeouts and 13 walks in 16 1/3 innings.
Manager Ron Gardenhire told Liriano of the club's decision during a meeting on Monday that also included pitching coach Rick Anderson, who will watch Liriano throw bullpen sessions on Thursday and Saturday.
"It's about being able to relax on the mound, and he knows that," Gardenhire said. "He actually says he has confidence, but he'll tell you straight out that when he gets men out there, he starts spinning 'em. So he's got to be able to control that. It's a work in progress. We're letting him take a step back and relax a little bit."
Gardenhire added that Liriano, who is on board with the decision to skip his start, also has to fix a few mechanical flaws.
"Andy and him are going to have long conversations," Gardenhire said. "They already are. As you can see, they're working on things, talking about things. He's wide open. He's had his struggles right now and he's wide open. He's searching for answers."


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Bird flu outbreak at Bernard Matthews turkey farms
Twenty thousands turkeys have been isolated on two Bernard Matthews breeding farms in East Anglia following an outbreak of a low-risk avian flu virus.
Government vets were called in following a significant drop in egg yield on Tuesday night. Tests have ruled out the deadly H5N1 strain and other highly pathogenic strains of H5 and H7.
It is unlikely the birds will be culled though it is unclear whether the eggs will be hatched to produce new turkey chicks for meat production.
The Health Protection Agency were informed but experts have decided the risks are so low that the 20 staff who work on the farms at Yaxham, near Dereham, Norfolk and Ubbeston, near Halesworth, Suffolk, do not require treatment with the anti-viral drug, Tamiflu.


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NEW YORK - The reunion of Brad Richards and Martin St. Louis is finally yielding some tangible results and there are signs the best is yet to come.
Things didnt go as smoothly as most thought they would upon the arrival of St. Louis at the NHL trade deadline in a rare swap of captains that saw Ryan Callahan shipped to the Tampa Bay Lightning.
Being put together on a line didnt bring immediate chemistry, but Richards and St. Louis have found a way get it done on separate units, sharing time only on the power play.
St. Louis needed some time to find his way and after recording only one goal and eight points in 19 games after the trade, the skilled forward is leading the Rangers in playoff scoring with two goals and six points heading into Game 6 on Tuesday at Wells Fargo Center.
Right behind him is Richards, who has two goals and five points in five games against the Flyers as the Rangers built a 3-2 edge in the best-of-seven match-up.
Those guys have got to share their experiences with our players and help them focus on the right thing, said Rangers head coach Alain Vigneault. The right things are not the outcomes, but the process.
Richards and St. Louis won a Stanley Cup together with the Tampa Bay Lightning back in 2004 so knowing the process is something that comes with the territory.
Its hard to believe it was 2007 the last time we were in the playoffs together, said Richards. It feels normal and its fun to be back talking hockey, talking about the playoffs and trying to figure out how to win.
Richards was asked if he has had to shoulder more of the leadership burden after the departure of Callahan?
Probably a little bit, yeah, said Richards, who has 30 goals and 83 points in 98 playoff games. Youve got to try to figure it out when a captain leaves the room, (whether) its by committee or whatever it is. There are certain responsibilities that he had that youve got to make sure are taken care of on and off the ice. Weve got good leadership all through the room and everybody has tried to take a piece of that.
As he has become more comfortable with the group, St. Louis has been more vocal and stepped into the leadership core as well.
Theyve achieved a lot in their careers. They take charge, said Rangers winger Mats Zuccarello. If we want to be successful, you need 20 guys riding the bus and theyre good at being leaders and getting everyone on the bus.
After a disappointing playoff last spring (one goal in 10 games) that ended with him in the press box as a healthy scratch, Richards was third in scoring during the regular season with 20 goals and 51 points and he has increased his production as the games have become more important.
Hes picked it up a notch here in the playoffs, said Rangers winger Carl Hagelin. Hes skating well, hes making plays and you can see that hes got a lot of confidence and hes really feeling it.
Richards is an alternate captain, but likely would have been the leading candidate to wear the C had Vigneault made the decision to name a new captain.
Hes always been vocal in the room. Hes been a leader in this league for a long time and he understands how to be a pro, said Rangers centre Derek Stepan. First of all, the hockey sense of Brad is very high. The highest IQ Ive played with. Knowing the game the way that he does, its good to be able to listen to him and soak up what he says.
The other thing is that he has experienced a lot in his career. Guys who have gone through it know what it takes to be successful in this league. Those are the guys that you look to and those are the guys that you lean on.
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