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Angiopoietin-1 (Ang-1) is essential for remodeling of the primitive vascular plexus and recruitment of mural cells during embryonic development. In the adult vasculature, Ang-1 can reduce plasma leakage in inflammation, but the mechanism of this action is not well understood. In the present study, we determined the magnitude and cellular mechanism of the antileak effect of Ang-1 in the airways of mice. Intravenous injection of bradykinin resulted in leakage of fluorescent microspheres (diameter 25–1,000 nm) from tracheal venules. The leakage peaked in 3–4 min and resolved by 10 min. High-resolution confocal microscopy revealed the presence of focal gaps at intercellular junctions of leaky venules. Genetically engineered Ang-1*, delivered systemically by adenoviral transduction of the liver, reduced leakage of 500-nm microspheres after bradykinin by 69%. The reduction in leakage coincided with a decrease in number and size of endothelial gaps. The proportion of venular surface occupied by endothelial gaps decreased 61%. Microsphere leakage correlated strongly with gap number and size (r2 = 0.89). Together the results suggest that Ang-1 reduces leakage from inflamed venules by restricting the number and size of gaps that form at endothelial cell junctions through effects on intracellular signaling, cytoskeleton, and junction-related molecules.
- confocal microscopy
- immunohistochemistry
- respiratory tract
- vascular permeability
angiopoietin-1 (Ang-1) is a ligand for Tie-2 tyrosine kinase receptors expressed on endothelial cells (9). Mutation of Ang-1 or its receptors is lethal during embryonic development because primitive vascular tubes do not evolve into mature functional blood vessels (11, 32). Ang-1/Tie-2 signaling is also thought to stabilize blood vessels through the recruitment of mural cells (32). Expression of Ang-1 ligand in mural cells and Tie-2 receptors in endothelial cells of quiescent vessels of the adult vasculature suggests a role for Ang-1 in maintenance of vascular stability (22, 30, 31, 44).
Plasma leakage, a hallmark of inflammation, has been attributed to the formation of intercellular gaps in the endothelium of postcapillary venules and collecting venules (24–26). Staining with silver nitrate or Ricinus communis agglutinin-I lectin, which mark sites of leakage, and ultrastructural studies have shown that endothelial gaps are focal openings resulting from transient separation of intercellular junctions (3, 16, 25, 33).
Ang-1 has a distinctive effect on plasma leakage. Dermal blood vessels in transgenic mice that overexpress Ang-1 in the skin are resistant to leakage induced by a variety of inflammatory mediators (35). Similar results have been obtained with COMP-Ang-1, a variant of Ang-1 that stimulates the formation of new blood vessels that are resistant to leakage (6). Moreover, systemic delivery of Ang-1 by adenoviral vector inhibits leakage induced by inflammatory stimuli (34). Ang-1 also has beneficial effects in models of endotoxic shock, retinopathy, and ischemia (28, 43, 45). The cellular mechanism of these effects of Ang-1 in vivo is unclear.
Studies of endothelial cells in vitro suggest that Ang-1 can decrease monolayer permeability, change the distribution and activation of junctional adhesion molecules, and stabilize cell-cell junctions. Ang-1 promotes recruitment of CD31 (platelet-endothelial cell adhesion molecule-1, PECAM-1) to junctions and decreases phosphorylation of CD31 and VE-cadherin (14, 18, 38). Ang-1 also reduces cytokine-induced expression of ICAM-1, VCAM-1, E-selectin, and tissue factor, all of which are implicated in the activation of endothelial cells and leukocyte adhesion and migration in inflammation (20, 21).
The present study sought to determine the magnitude and cellular mechanism of the antileak effect of Ang-1 in a mouse model of airway inflammation. Adult mice were injected intravenously with recombinant adenovirus encoding a genetically engineered version of angiopoietin-1 called Ang-1* (34). Adenovirally transduced hepatocytes generated systemic levels of Ang-1* in the bloodstream. Three days later the tracheal vasculature was challenged by intravenous injection of bradykinin (26). The dose response and time course of bradykinin-induced leakage were assessed by measuring extravasation of Evans blue dye. Leaky vessels were identified, and pore cutoff size was determined by quantifying extravasated fluorescent microspheres. The integrity of endothelial cell junctions and formation of endothelial gaps were evaluated by examining immunohistochemically stained tracheal whole mounts by confocal microscopy.
Animals and Treatment
The effect of Ang-1* on vascular leakage was examined in pathogen-free FVB/n 8-wk-old male mice (Charles River, Hollister, CA). Mice were anesthetized by intramuscular injection of ketamine (87 mg/kg) and xylazine (13 mg/kg), and 109 plaque-forming units of Ang-1*-expressing adenovirus were injected into a jugular vein (Ad-Ang-1* group) (34). At 3 days after the injection of Ad-Ang-1*, serum concentrations of Ang-1* were 1–15 μg/ml in all mice, consistent with published observations (34). Age-matched control FVB/n mice received an injection of adenovirus expressing green fluorescent protein (GFP; Ad-GFP group) or nothing. GFP gene expression in hepatocytes, which were the main target of adenoviral transduction, was confirmed by fluorescence microscopy (34). Ang-1* was not detected in blood of the control mice.
Three days after the injection of adenovirus, mice were challenged by intravenous injection of bradykinin (Sigma, St. Louis, MO) at a dose of 0.01, 0.1, 1, 10, 20, or 50 mg/kg diluted in 100 μl of 0.9% NaCl; control mice received 100 μl of 0.9% NaCl. All experiments were approved by the University of California, San Francisco Institutional Animal Care and Use Committee.
Measurement of Evans Blue Leakage
Mice used to measure the dose response and time course of bradykinin-induced plasma leakage of Evans blue dye were anesthetized with ketamine and xylazine. Evans blue (30 mg/kg in 100 μl of PBS; Electron Microscopy Sciences, Fort Washington, PA) was injected into a femoral vein, followed 2 min later by injection of bradykinin (4, 35). Five to sixty minutes after the Evans blue injection, the chest was opened, the atria were removed to create an exit route, the left ventricle was incised, an 18-gauge cannula was inserted through the ventricle into the ascending aorta, and then intravascular dye was flushed from the bloodstream by perfusion of 1% paraformaldehyde (PFA) in citrate buffer (0.05 M, pH 3.5) at a pressure of 120 mmHg for 3 min. Completeness of the perfusion was confirmed by uniform clearing of blood from the trachea and other organs. The effectiveness of this approach for rinsing blood from the vasculature was previously verified by infusion of a labeled lectin that documented the absence of blood by binding uniformly to tracheal blood vessels (33). Tracheas were removed, weighed, and incubated in formamide overnight at 67°C (Sigma). Extracted Evans blue was measured with a spectrophotometer at 620 nm and expressed as nanograms of dye per milligram of tissue wet weight.
Localization and Measurement of Microsphere Extravasation
Sites of leakage in tracheal blood vessels were marked by extravasated fluorescent polymer microspheres ranging in diameter from 25 to 1,000 nm (20 μl iv; Duke Scientific, Palo Alto, CA) (25). Because the microsphere preparations had a uniform weight per volume relationship, the number of microspheres in 20 μl varied inversely with diameter: 2 × 1013 (25 nm), 4 × 1011 (100 nm), 3 × 109 (500 nm), and 3 × 108 (1,000 nm). Mice anesthetized with Nembutal (40 mg/kg ip) received an intravenous injection of bradykinin (1 or 10 mg/kg) or saline and 0–9 min later an injection of microspheres. One minute thereafter, intravascular microspheres were removed from the bloodstream by vascular perfusion of 1% PFA in PBS, following the same approach as for Evans blue.
Tracheas were removed and fixed for an additional 2 h, and then the vasculature was stained for CD31 immunoreactivity (2). Tracheal whole mounts were incubated for 1 h in 5% normal goat serum (Jackson ImmunoResearch, West Grove, PA) in PBS containing 0.3% Triton X-100 (PBS-plus) and then incubated overnight at 4°C with hamster anti-mouse CD31 (clone 2H8; Chemicon, Temecula, CA; 1:1,000 in PBS-plus) and/or rat anti-mouse CD31 (MEC13.3; Pharmingen, San Diego, CA; 1:500 in PBS-plus). The next day, tracheas were washed several times with PBS-plus and incubated for 4 h at room temperature with FITC- or Cy3-conjugated goat anti-hamster and/or anti-rat IgG (Jackson ImmunoResearch; 1:200 in PBS-plus). Tracheas were rinsed several times in PBS-plus, flattened, and mounted with the luminal surface facing up in Vectashield (Vector Laboratories, Burlingame, CA).
CD31-stained tracheal whole mounts were examined with a Zeiss Axiophot fluorescence microscope (×10 objective, ×1 Optovar, tissue region 960 × 1,280 μm) equipped with fluorescence filters and a low-light, externally cooled, three-chip charge-coupled device camera (480 × 640 pixel RGB color images; CoolCam, SciMeasure Analytical Systems, Atlanta, GA). Area density of extravasated microspheres (proportion of total pixels) was measured on digital images of regions of tracheal mucosa between five to eight cartilage rings where venules and postcapillary venules were most abundant. Camera settings were uniformly applied to images from all groups in each experiment (19). RGB color images were converted into 8-bit gray scale, and then ImageJ software (http://rsb.info.nih.gov/ij) was used to determine the proportion of pixels having a fluorescence intensity equal to or greater than specific threshold values that maximized the microsphere signal and minimized tissue autofluorescence. Thresholds were found empirically to be ≥40 for green or red 100-nm microspheres or ≥20 for blue 500-nm microspheres (fluorescence intensity range: 0–255 in 8-bit images). Area densities of microspheres in a 205,920-pixel region of interest in each image were expressed as the percentage of pixels with a brightness equal to or greater than the threshold. An average area density value was calculated for all images from each mouse (3–4 mice/group).
Confocal Microscopic Imaging of Endothelial Gaps
For confocal microscopic studies of endothelial gaps, mice were anesthetized with Nembutal and then received an intravenous injection of bradykinin (1 mg/kg) or saline. Fluorescent microspheres (100 or 500 nm) were injected at the time of the bradykinin injection or from 1 to 9 min later. Fixation by vascular perfusion of 1% PFA began 1 min after injection of microspheres, thereby standardizing the microsphere circulation time in all experiments. Tracheas were stained for CD31 immunoreactivity with both hamster and rat anti-mouse CD31 antibodies, rinsed in PBS-plus, mounted in Vectashield, and examined with a Zeiss LSM-510 confocal microscope with argon, helium-neon, and UV lasers.
The two anti-CD31 antibodies labeled endothelial cell junctions in arteries and veins but played separate roles in the identification of endothelial gaps because they labeled different populations of CD31 molecules in endothelial cells of tracheal capillaries and venules. The hamster antibody labeled CD31 at intercellular junctions, and the rat antibody labeled a population of CD31 molecules more uniformly distributed in the plasma membrane. We took advantage of these properties to distinguish unstained intercellular gaps from regions of nonjunctional plasma membrane, which stained with the rat antibody but not the hamster antibody. When the hamster antibody was used alone, cell junctions were labeled but intercellular gaps could not be distinguished from nonjunctional regions of plasma membrane because neither was stained. Addition of the rat antibody solved that problem. By uniformly labeling endothelial cell plasma membranes, the rat antibody also made measurements of vessel diameter more accurate.
Measurement of Endothelial Gap Number and Size
Unlike measurements of microsphere area density, the number and size of endothelial gaps were determined in confocal microscopic images of tracheal venules in mice injected with bradykinin (1 mg/kg) or saline, followed 1 min later by 500-nm microspheres. The vasculature was perfused with fixative 2 min after the bradykinin, and tracheal whole mounts were stained with the two anti-CD31 antibodies. Confocal images were obtained from the tracheas of four or five mice in each of four groups: two Ad-Ang-1* groups and two Ad-GFP groups, one of each challenged with bradykinin and the other with saline. A total of 15–17 confocal image projections were examined in each group (×40 oil objective, ×3 zoom). Each projection represented a stack of 28–64 optical sections, each 0.39–0.58 μm in thickness, through the entire vessel wall. Scanning parameters were adjusted for each image to optimize gap identification. At this high magnification, intercellular gaps were clearly visible at endothelial cell borders marked by the hamster CD31 antibody.
All optical sections in the image stacks were examined to identify and encircle each gap with the Line Drawing and Measuring function of the Zeiss LSM Image Browser software (http://18.104.22.168/LSMContent.htm). Gap borders thus drawn were used to determine the number, perimeter, and area of gaps, which in turn were used to calculate the gap minor and major axes. The number of gaps was expressed per millimeter of venule midline length. Venular length, measured with a digitizing tablet (Digi-Pad; GTCO CalComp, Scottsdale, AZ), rather than surface area was used as the reference because prolonged exposure to Ang-1* can result in circumferential enlargement of venules due to endothelial cell proliferation (2, 7, 36). In our experiments, the enlargement was minimal at 3 days after Ad-Ang-1*.
Perimeters of endothelial gaps in confocal images were measured with the Closed Polyline Overlay measuring tool of the Zeiss LSM Image Browser software. Gap areas were calculated from the number of pixels contained within these perimeters.
The minor axis of gaps was calculated from gap perimeter (P) and area (A), based on an approximation formula for the circumference of an ellipse (15)
The major axis of gaps was calculated from gap area, perimeter, and minor axis (15)
The proportion of venular surface occupied by gaps in each image projection was calculated from the total area of gaps and vessel surface area by treating venules as cylinders. Average vessel diameter and length were measured on each image projection, and endothelial surface area was calculated with the formula for surface area of a cylinder (2πrl, where r is the radius and l is the length). Fractional gap area was expressed as square micrometers of gap openings per square millimeter of venular surface.
Statistical Analysis
Values are expressed as means ± SE. Experimental groups consisted of three to nine mice unless otherwise indicated. The significance of differences among groups was assessed by ANOVA followed by the Bonferroni-Dunn test for multiple comparisons. The relationship between area density of extravasated microspheres and proportion of venular surface occupied by gaps was assessed by linear regression. Differences between groups were considered significant when P < 0.05.
Bradykinin-Induced Leakage of Evans Blue
Before examining the effect of Ang-1* on vascular leakiness, we examined the dose response of bradykinin-induced leakage of Evans blue in the mouse trachea. Mice received Evans blue followed 2 min later by an injection of saline or bradykinin. Dye accumulation in the trachea was measured 30 min after injection of Evans blue and expressed as a percentage of the baseline value. After injection of saline, the baseline concentration of Evans blue at 30 min was 21.2 ± 3.5 ng/mg trachea (Fig. 1A). Doses of bradykinin from 0.01 to 0.1 mg/kg did not increase the value above this baseline, but 1 mg/kg caused a 38% increase and 10 mg/kg produced an 86% increase (Fig. 1A). Larger doses of bradykinin (20 or 50 mg/kg) produced somewhat more leakage (Fig. 1A) but were lethal in some mice.
We next examined the time course of the leakage. Injection of Evans blue was followed 2 min later by an injection of bradykinin (10 mg/kg) or saline. The dye was flushed from the vasculature by perfusion of acidified fixative 5, 30, or 60 min after the dye injection (Fig. 1B). Saline-injected groups provided a reference for baseline Evans blue accumulation at the three time points. At 5 min after bradykinin challenge, tracheal Evans blue content was 91% higher than the 5-min baseline value. Dye content was still increased at 30 min (123% above the 30-min baseline) but decreased thereafter, and at 60 min was only 25% above the 60-min baseline (Fig. 1B).
Bradykinin-Induced Leakage of Fluorescent Microspheres
Evans blue dye studies helped to define the dose response and time course of leakage after bradykinin, but high baseline values made the approach relatively insensitive for monitoring changes in blood vessel leakiness in the trachea. Therefore, we used 25- to 1,000-nm fluorescent microspheres as tracers in subsequent experiments. Particles of this size do not cross the endothelium of tracheal vessels in the absence of increased permeability, and little extravasation occurs under baseline conditions. Extravasated microspheres had the additional attribute of marking sites of leakage and providing an estimate of the pore cutoff size of the leaky sites.
Location and pore size of leakage sites.
Microspheres were injected 1 min after saline or bradykinin (10 mg/kg) and were removed from the bloodstream by vascular perfusion 4 min later. Each mouse received microspheres of only one size. Extravasated microspheres were rare after saline challenge but were abundant after bradykinin. Most extravasated microspheres, regardless of the size injected, were located in the wall of venules (Fig. 2, A–D). The number of extravasated microspheres in venular walls varied with the size injected (Fig. 2, A–D). The smallest microspheres (25 nm) were most abundant (Fig. 2A), consistent with their greater extravasation propensity and abundance due to logfold differences in number injected (see materials and methods). Even the largest microspheres extravasated, indicating the presence of openings as large as 1,000 nm (Fig. 2D). CD31-positive leukocytes were present in some venules after bradykinin (Fig. 2E). Few microspheres extravasated from arterioles or capillaries, as found in previous studies (25, 26).
Time course of leakage.
To determine the time course of microsphere leakage, 100-nm microspheres were injected with bradykinin at time 0 or 1–9 min thereafter. In all cases, the microspheres circulated for 1 min and were then flushed from the bloodstream by vascular perfusion of fixative. Baseline leakage was assessed after injection of saline followed by microspheres and 1 min later by perfusion of fixative. Area density measurements showed that accumulation of 100-nm microspheres after a 10 mg/kg dose of bradykinin was 3-fold greater than baseline during minute 1, 10-fold greater during minute 2, and 15-fold greater during minute 3 and was back to baseline by minute 10 (Fig. 3, A–E and G). Bradykinin at 1 mg/kg resulted in a similar amount of leakage during minute 2 (Fig. 3, F and H). Because microsphere circulation time was held constant, where perfusion times ranged from 1 to 10 min after bradykinin injection, the amount of extravasated microspheres reflected leakage rather than clearance of microspheres from the circulation.
Magnitude of Ang-1* Effect on Blood Vessel Leakiness
Effect of Ang-1* on Evans blue leakage.
On the basis of data for the dose response of bradykinin and the time course of Evans blue leakage, we next examined the antileak effect of Ang-1*. Mice received Ad-Ang-1* or Ad-GFP, and 3 days later were challenged with saline or bradykinin (10 mg/kg) followed by Evans blue that circulated for 30 min. In the Ad-GFP group bradykinin increased tracheal dye accumulation by 60% above the saline baseline (Fig. 4A), but in the Ad-Ang-1* group bradykinin produced no detectable increase above baseline (Fig. 4A). In the absence of the bradykinin stimulus, baseline leakage over 30 min was about the same in both groups (Fig. 4A).
Effect of Ang-1* on microsphere leakage.
Multiple factors, including blood flow, vascular surface area, and endothelial barrier function, determine the amount of Evans blue accumulation in tissues. Increased tissue content of a tracer such as Evans blue that can cross the normal endothelium is not proof of increased endothelial permeability. Therefore, we repeated the Ang-1* studies with a particulate tracer, fluorescent 500-nm microspheres, which are too large to extravasate from tracheal vessels without a defect in endothelial barrier function.
With the goal of determining whether the antileak effect of Ang-1* involves a permeability decrease consistent with restriction of endothelial gap formation, we asked whether Ang-1* reduces leakage of 500-nm microspheres. Mice received Ad-Ang-1* or Ad-GFP and 3 days later were challenged with bradykinin (1 mg/kg) or saline. Leakage from tracheal blood vessels was assessed by quantifying the accumulation of 500-nm microspheres. Leakage of microspheres at 2 min after bradykinin was 39 times the saline baseline value in the Ad-GFP group but 12 times baseline in the Ad-Ang-1* group, reflecting a 69% reduction compared with the Ad-GFP group (Fig. 4B). This change could be explained by a reduction in gap number, size, or both.
Endothelial Gap Formation After Bradykinin
The results of studies with fluorescent microspheres led us to learn more about changes in the endothelium of venules after bradykinin challenge. In control mice, most endothelial cell borders of tracheal venules were smooth (Fig. 5, A and A′), and few or no endothelial gaps were visible by confocal microscopy after immunohistochemical staining with the two anti-CD31 antibodies. Because the hamster anti-mouse CD31 antibody stained endothelial cell borders and the rat anti-mouse CD31 antibody uniformly stained nonjunctional endothelial cell plasma membranes, gaps could be seen as unstained regions between endothelial cells. Similar observations were made when endothelial cell junctions were stained for VE-cadherin or ZO-1 immunoreactivity (data not shown). At 1 min after bradykinin (1 mg/kg), gaps were visible between many endothelial cells of venules (Fig. 5B). The number and size of gaps increased between 1 and 4 min (Fig. 5, B and C). Extravasated microspheres were present near some gaps at 1 min (Fig. 5B) and were more abundant and widely distributed on venules at 4 min (Fig. 5C). Fingerlike processes (filopodia) partitioned some of the larger gaps (Fig. 5, C and C′). At 10 min, gaps and extravasated microspheres were rare, and endothelial cell borders again had a smooth contour with few filopodia (Fig. 5D).
Effect of Ang-1* on Gap Number and Size
Consistent with its effect on microsphere leakage, Ang-1* reduced both the number and size of endothelial gaps in tracheal venules after bradykinin challenge. In the Ad-GFP group examined 2 min after bradykinin (1 mg/kg), gaps of various size were visible at the borders of many endothelial cells and 500-nm microspheres were abundant in the wall of venules (Fig. 6, A–C). Measurements showed on average 836 gaps/mm venule length (Fig. 6G). By comparison, endothelial gaps in the Ad-Ang-1* group were 38% less numerous (516 gaps/mm) and were accompanied by fewer extravasated microspheres (Fig. 6, D–G). Gaps were rare in venules of saline-challenged mice in either group (Fig. 6G).
In addition, gaps were 38% smaller in the Ad-Ang-1* group than in the Ad-GFP group (gap area: 0.45 vs. 0.73 μm2; Fig. 6H). The mean major axis of gaps in the Ad-Ang-1* group was reduced 22% (0.76 vs. 0.98 μm; Fig. 6I). Mean minor axis was 20% smaller in the Ad-Ang-1* group (0.16 vs. 0.20 μm; Fig. 6I). The aspect ratio (minor axis as a proportion of major axis) was almost identical in the two groups (0.22 vs. 0.23), indicating that gaps tended to be highly elongated regardless of treatment. Together, these measurements indicate a 61% reduction in venular surface area occupied by gaps after Ad-Ang-1* (0.28% of endothelial surface) compared with the value in the Ad-GFP group (0.71% of endothelial surface; Fig. 6J). Corresponding values for saline-challenged mice were 0.02% and 0.03% (Fig. 6J), which was at the limit of gap detection.
Comparison by linear regression showed that the proportion of vessel wall occupied by gaps had a significant positive correlation with the amount of extravasated 500-nm microspheres on venules (P < 0.0001, correlation coefficient r2 = 0.89; Fig. 7).
The present study sought to determine the magnitude and cellular mechanism of the antileak effect of Ang-1 in airway inflammation. Our approach was to inject mice intravenously with adenovirus encoding Ang-1* or GFP, which transduced hepatocytes and produced systemic levels of the protein in the bloodstream. Three days later we examined the leakiness of blood vessels in the tracheal mucosa after intravenous challenge with bradykinin. We found that Ang-1* reduced the leakage of Evans blue to baseline and the leakage of 500-nm fluorescent microspheres by 69%. Confocal microscopic analysis revealed that, after Ang-1*, gaps between endothelial cells of mucosal venules were smaller and less numerous. Together, these changes decreased by 61% the fraction of venular surface occupied by endothelial gaps after bradykinin challenge. Ang-1* thus reduces leakage produced by inflammatory mediators in part by decreasing the number and size of gaps that form at intercellular junctions. This antileak action of Ang-1* is probably not limited to the response to bradykinin because similar effects are found with a wide range of inflammatory stimuli (14, 23, 29, 34, 35, 38).
Plasma Leakage in Airways
Bradykinin, a member of the kinin family of vasodilator peptides, was used as the stimulus to test the antileak effect of Ang-1* because it is known to produce rapid but transient plasma extravasation from venules in the respiratory tract (5). This action of bradykinin is mediated via B2-receptor signaling in endothelial cells. Leakage is restricted to postcapillary venules and is thought to result from the formation of gaps between endothelial cells (1, 12, 17).
To optimize the model for studying the effects of Ang-1*, we first determined the dose response and time course of Evans blue accumulation in the mouse trachea after bradykinin challenge. An intravenous dose of 10 mg/kg of bradykinin nearly doubled the Evans blue content of the trachea. Higher doses produced somewhat more leakage but were lethal in some mice. Dye accumulation was increased at 5 min, still elevated at 30 min, and almost back to baseline by 60 min.
High baseline Evans blue values indicated that appreciable dye extravasated from tracheal blood vessels even without bradykinin. Therefore, we sought to confirm that bradykinin did indeed increase permeability in this system. For this purpose we used fluorescent microspheres that were too large to extravasate without a change in endothelial barrier function. Baseline leakage of 25- to 1,000-nm microspheres was very low. Bradykinin challenge was followed by extensive leakage of microspheres, which was transient, peaked at ∼3 min, and returned to the near-zero baseline by 10 min. By keeping the microsphere circulation time constant at 1 min, it was possible to distinguish transience of leakage from rapid clearance of microspheres from the bloodstream. Because, unlike Evans blue, 500-nm microspheres do not leak from normal tracheal vessels, the increase in microsphere leakage after bradykinin (∼39-fold) was much larger than for Evans blue (∼1.6-fold).
Unlike extravasated Evans blue that moved throughout the tissue, most extravasated microspheres were trapped in the wall of leaky vessels by the endothelial/pericyte basement membrane. This is a well-documented property of particulate tracers (25, 26). Extravasated microspheres thus marked the leaky vessels, which were easily identified as postcapillary venules or collecting venules in the simple architecture of the tracheal microcirculation. Restriction of the leakage to this segment of the vasculature led us to explore the role of endothelial gaps (24–26).
Role of Endothelial Gaps in Plasma Leakage
Multiple mechanisms have been proposed, discussed, and debated to explain the movement of macromolecules across the vascular endothelium (8, 13, 26, 27). Intercellular gaps, transcellular holes, fused endothelial vesicles, vesiculovacuolar organelles, transcytosis, and fenestrations have all been considered potential contributors. The classic studies of Majno and Palade (24) implicated endothelial gaps in plasma leakage from inflamed venules. Results of many subsequent studies are consistent with that view (1, 3, 16, 17). The presence of endothelial gaps at sites of particulate tracer leakage and the extravasation of tracers from inflamed vessels after fixation provide further evidence for openings in the endothelium that permit extravasation driven by hydraulic pressure gradients rather than transcellular transport (26). The present studies showed that microspheres from 25 to 1,000 nm in diameter extravasate after bradykinin challenge. As endothelial gaps are reported to have an average size of ∼500 nm (26), they appear to be a reasonable pathway for extravasation of microspheres in the system we studied.
A conventional method for studying endothelial gaps involves electron microscopy (1, 3, 16, 17, 24). This approach has the attribute of high resolution but may be limited by the small regions sampled. To explore the involvement of endothelial gaps in our system, we developed a confocal microscopic approach for determining the number and size of endothelial gaps in leaky venules of mouse airways. For this purpose, tracheal whole mounts were stained by immunohistochemistry using two different antibodies to the endothelial cell junctional molecule CD31/PECAM-1. Because the hamster anti-mouse CD31 antibody selectively labeled endothelial cell junctions and the rat anti-mouse CD31 antibody uniformly stained nonjunctional endothelial cell plasma membranes, gaps could be seen as unstained holes surrounded by intensely stained cell junctions. Without the rat anti-mouse CD31 antibody, gaps could not always be distinguished from nonjunctional regions of plasma membrane.
Using this approach, we examined endothelial gap formation in bradykinin-induced leakage. Within only 1 min of bradykinin injection, gaps were visible at focal regions of intercellular junctions marked by CD31 staining where endothelial cells appeared to retract from one another. Extravasated microspheres accumulated near gaps.
With its potential for broad sampling, the confocal microscopic approach was more efficient for determining the size, shape, number, and distribution of endothelial gaps than transmission or scanning electron microscopy as used in previous studies (3, 16). The lower resolution of confocal microscopy made gap size measurements not as accurate as with electron microscopy, and counts of gap numbers probably underestimated the actual values because gaps smaller than ∼100 nm may not have been visible. However, the confocal microscopic approach made it possible to measure a large population of gaps with a precision adequate to detect changes in gap number and size under the conditions of our experiments.
After bradykinin, endothelial gaps in airway venules tended to be slitlike, with an average length (980 nm) fivefold the width (200 nm). By 10 min, gaps were no longer visible, and junctions appeared normal. These findings are consistent with what is known about the time course of endothelial gap formation and leakage after exposure to mediators in several models of acute inflammation (3, 16, 25, 26).
Effect of Ang-1* on Number and Size of Endothelial Gaps
Ang-1* completely blocked the accumulation of Evans blue dye and significantly reduced the leakage of 500-nm microspheres in the trachea after bradykinin challenge. Similar results have been obtained in mouse skin, in which adenoviral Ang-1* or transgenically overexpressed Ang-1 inhibits Evans blue leakage induced by local application of mustard oil, VEGF, serotonin, or platelet-activating factor (34, 35). In cultured endothelial cell monolayers, Ang-1 reduces baseline permeability and response to bradykinin, histamine, VEGF, thrombin, and TNF-α (14, 23, 29, 38). It is unknown whether Ang-1 modifies bradykinin receptor expression or its downstream signaling, but the antileakage action against a wide range of mediators favors a more generalized effect on endothelial barrier function.
Confocal microscopic studies revealed that Ang-1* reduced both the number (38% reduction) and the size (38% reduction) of endothelial gaps that formed in airway venules after bradykinin. Together these changes represented a 61% decrease in the area of venular surface occupied by endothelial gaps. This change fit with the 69% decrease in extravasation of 500-nm microspheres.
The restriction in endothelial gap formation by Ang-1* is likely to involve changes in cell-cell junctional molecules. Ang-1 increases expression of the tight junction molecule occludin, decreases phosphorylation of CD31 and VE-cadherin, and increases CD31 at endothelial cell-cell junctions, which may contribute to improved barrier function (14, 18). Ang-1 increases the interaction of VE-cadherin and β-catenin at cell-cell junctions and the activation of PKC-β associated with VEGF-induced increased monolayer permeability (38). Ang-1 also inhibits thrombin-induced activation of PKC-ζ (23).
Further studies will be needed to determine whether the effects on endothelial gaps of adenovirally delivered Ang-1* can be reproduced by administering recombinant Ang-1 in vivo, where Ang-1 stability and pharmacokinetic and pharmacodynamic issues come into play. Future experiments should also address whether Ang-1* limits gap number and size by limiting gap opening or speeding gap closure. Indeed, our studies of gap formation focused on the first minutes after bradykinin with the thought that Ang-1 may hasten gap closure, but the speed of gap opening and closure made this a challenging question to address in vivo with current methods.
Under baseline conditions, junctional molecule signaling helps maintain the quiescent phenotype of endothelial cells (10, 37). Inflammatory mediators change this in part through actions on the cytoskeleton linked to junctional molecules (39, 40). Contraction of cytoskeletal elements seems not to be essential for gap formation. Rather, gaps form by elastic recoil of endothelial cell borders when junctional and cytoskeletal attachments are focally weakened or disrupted (39, 41).
Can changes in number and size of endothelial gaps explain all effects of bradykinin and Ad-Ang-1* on microsphere extravasation? Although participation of vesiculovacuolar organelles or transcellular holes near endothelial cell borders cannot be excluded, Ad-Ang-1* caused similar reductions in amount of venular surface occupied by gaps and in leakage of 500-nm microspheres. Gap number and size, expressed as a proportion of endothelial surface under the various conditions, were strongly correlated with amount of microsphere extravasation (correlation coefficient r2 = 0.89). Furthermore, the proportion of venular surface occupied by gaps in our study is in the range of earlier estimates based on different methods (16, 25, 26). Determination of the quantitative relationship between gap number/size and amount of microsphere extravasation will require more information on the contribution of Starling forces to transvascular fluid flux in our system.
The inhibitory action of Ang-1 on endothelial gap formation and plasma leakage in the airways has potential clinical significance because mucosal edema is a common feature of inflammatory diseases such as asthma and chronic bronchitis (5, 42). Ang-1-like therapeutic agents may be clinically beneficial by reducing airflow obstruction associated with airway edema.
This work was supported in part by National Heart, Lung, and Blood Institute Grants HL-59157 and HL-24136.
We thank Drs. George Yancopoulos, Ella Ioffe, and Donna Hylton (Regeneron) for generously supplying the adenoviral vectors and measuring serum Ang-1*.
Present address of F. Baffert: Novartis, Basel, Switzerland (e-mail: fabienne.baffert{at}novartis.com).
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 © 2006 by the American Physiological Society


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Adak, Alaska (PressExposure) October 12, 2006 -- Are you the type of guy who can eat and eat and still not put on the weight you want? Do you push yourself harder and harder in the gym and you're not noticing any changes to the size of your body? Are you finding yourself eating more throughout the day in the hopes that those extra calories will pack on the muscle size you're looking for? Well I have an easier solution to your problems and the answer is weight gainer [http://worldofsupplements.com/].
Not everyone was blessed with the genetics of a professional bodybuilder, or a professional athlete. For those of us who do not have the ability to pack on the muscle with a normal diet, or a normal protein powder, we have to rely on other sources. The best thing that can be done for the ones who struggle to put on weight is to begin supplementing with a weight gainer [http://worldofsupplements.com/]. Weight Gainer is an easy addition to any diet and exercise regimen. It's as easy as adding a shake or two throughout the days, especially in times when the muscles are depleted of nutrients so that you can achieve optimal growth.
Weight gainer can be found at any nutrition store and is usually purchased in powder form, just like your protein powder. The nutrients that are found in Weight gainer are ones that have been proven to slow down the body's metabolism and allow the muscles to really feed off of the nutrients that are being provided. Weight gainer is high in calories and is packed full of carbohydrates (usually waxy maize starch that helps force the proteins into the muscles) protein and all of the amino acids that are required for optimal growth.
Some of your high end Weight gainer can also contain high values of creatine to ensure that retention of nutrients is being held in the muscle bellies.Weight gainer is the answer to all of your muscle building needs. With all the right nutrients packed into one shake how can you deny the potential of adding Weight gainer to your diet? With weight gainer [http://worldofsupplements.com/] all of your goals can be achieved!

subset: commoncrawl


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NU Looks to Establish Ground Game
LINCOLN The Nebraska football team held its final full-scale workout of the week Thursday with a two-hour practice inside Memorial Stadium.
Head Coach Bill Callahan reiterated the importance of Nebraska establishing a running game this Saturday in its final home game against Kansas State.
"Every game that we go into, we try and establish a running game and have a balance in our attack," Callahan said. "Sometimes we're fortunate to do that, and sometimes we're not, but we want to be able to do that Saturday. We want to be able to establish the run and do what we need to do up front. There are certain things we've schemed and planned that we want to attack their defense with, and we're confident that we can do that. Let's hope it all comes to (fruition)."
Callahan also commended this year's senior class Thursday, noting its work ethic and leadership qualities in helping to build team unity in 2005.
"They've meant a lot to me, just because of the hard work, effort, determination and dedication they've put into this program," Callahan said. "They're a great group of guys. They want to win, and that's the thing that impresses me most about this group. Day-in and day-out, they're great to be around. They're fun in practice and work hard. More importantly, they're great guys off the field. They've been pivotal this year in helping us change some things in terms of our team unity. They've been instrumental in the overall unity of our team. They've been a real positive force."
On the injury front, Callahan reported that left tackle Cornealius Fuamatu-Thomas has a "great chance" of being ready to play Saturday after missing practice time the last two weeks due to injury.
The Huskers will hold a short walkthrough Friday afternoon at Memorial Stadium as they conclude their preparations for Saturday's 1 p.m. matchup with Kansas State.
NOTE: Nebraska hosted a familiar visitor during practice, as Josh Bullocks, former Husker free safety and twin brother of current NU strong safety Daniel Bullocks, was on hand Thursday. Josh, the starting strong safety for the NFL's New Orleans Saints, is with family in Lincoln during his bye week to help celebrate Senior Day with Daniel Saturday.


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Israel News
Middle East
American Politics
Edition française
Green Israel
Battir cultural landscape site 370.
(photo credit:Wikimedia Commons)
JERUSALEM - Israel's highest court on Thursday urged defense officials to reroute a barrier planned for construction on ancient farmland at a West Bank Palestinian village.The latest decision followed a rare appeal involving support from an Israeli government environmental agency for a Palestinian challenge.
By DANIEL K. EISENBUD
By LAHAV HARKOV
Sponsored Content
By DANIEL K. EISENBUD
By JEREMY SHARON
Please insert a valid email address


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We have compiled a list of Best Reference Books on Fire Dynamics Subject. These books are used by students of top universities, institutes and colleges.
Here is the full list of best reference books on Fire Dynamics.
|1. "An Introduction to Fire Dynamics" by Dougal Drysdale
Book Review: This is a very good textbook for fire safety engineers, researchers, and graduate and under graduate students who are involved in practicing fire dynamics. The book presents up-to-date information that incorporates the latest research and experimental data. The book addresses the fundamentals of fire science and fire dynamics and also provides the scientific background that is needed for the development of fire safety engineering as a profession. The book contains experimental data that is needed for the understanding of fire behavior of materials and contains numerous problems along with solutions .
|2. "Enclosure Fire Dynamics" by Bjorn Karlsson and James G.Quintiere
Book Review: This book helps in filling the gap that is involved in detailed understanding of scientific aspects of enclosure fires. The book provides complete description of enclosure fires and demonstrates how the fire outbreak in the compartment causes environmental changes. The authors provide a very nice and clear presentation of the mechanisms that control the enclosure files and develop simple analytical relationships that are useful in designing buildings for fire safety. The authors also explain the derivation of engineering equations from first principles. This book is very useful for fire protection engineers, researchers and investigators.
|3. "Fundamentals of Fire Phenomena" by James G.Quintiere
Book Review: The book describes all the scientific principles that are involved behind the fire behavior. This book also combines the principles of physics, chemistry, heat transfer, mass transfer and fluid dynamics that is necessary to understand the fundamentals of fire phenomena. The book also covers thermo chemistry that includes mixtures and chemical reactions. The book also discusses premixed flames and spontaneous ignition. This is a very good reference book for practicing engineers in safety and forensic analysis. The book also describes theoretical bases for empirical aspects.
|4. "Tunnel Fire Dynamics" by Anders Lonnermark
|5. "Tropical Fire Ecology_Climate Change, Land Use And Ecosystem Dynamics" by Cochrane
|6. "Fire Dynamics Tools (FDTs) Quantitative Fire Hazard Analysis Methods for the U.S. Nuclear Regulatory Commission Fire Protection Inspection Program" by NRC Staff: United States Nuclear Regulat
|7. "Fire Dynamics with MyFireKit " by Gregory E. Gorbett
|8. "Fire and Vegetation Dynamics: Studies from the North American Boreal Forest (Cambridge Studies in Ecology) " by Edward A. Johnson
|9. "Fire Dynamics Simulator (Version 4) – Technical Reference Guide" by nist
|10. "Fire Dynamics (Brady Fire)" by Gregory E. Gorbett
If any more book needs to be added to the list of best books on Fire Dynamics Subject, please let us know.
Sanfoundry Global Education & Learning Series – Best Reference Books!


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Process Technician
Gather and analyze information about production processes.
Directs flight path of remotely-piloted photo reconnaissance or target aircraft along predetermined flight course from command center in guide aircraft or ground station: Operates aircraft controls from command center by remote radio and closed circuit video transmission to direct aircraft to take off, follow flight path, and land, utilizing knowledge of aircraft navigation and aircraft flight characteristics. Analyzes performance data and prepares reports covering airframe, guidance, power plant, and mechanical controls of aircraft after ground or flight testing. Confers with engineering, inspecting, and manufacturing personnel concerning success of flight tested aircraft or need for corrective action. May assist with preflight inspection of aircraft. May direct flight path of guided missiles, utilizing remote computerized controls, and be designated Missile-Control Pilot.


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New versions of antivirus programs come with involuntary refurbish features. You only bond to a Internet, start a antivirus module as well as click upon a refurbish button. In box we have been lost, we can deliberate with your discriminate correct technician.
Martin Green is an Online Technical Support Executive for a site http://www.iyogi.ca who provides minute report upon Personal Support, Personal Personal Repair Services, Online Technical Support, Personal Repair, 24/7 Support, Antivirus await etc.


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The owners of Heart of Gold take their jewelry very seriously. No fancy mall location for them, just a cozy, friendly neighborhood storefront.
Their focus is on beautiful de...
All reviews seem positive
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Fernando Aguerre — the man who took happiness off an Argentine beach and spread it Latin-style to the coolest stores and showrooms in California and the world — is, at the moment, not wholly happy. He's in about the most pain of his life, suffering a herniated disc caused by lifting two antique balsa-wood surfboards in a Peruvian airport when his driver was late.
About this column
Steve Chapple's Intellectual Capital covers game-changing people, ideas and perspectives. His columns, which appear twice a month in the U-T's Sunday Business section, are entertaining visits with some of the smartest minds around San Diego. These are people ahead of the wave, with a wise eye on the past, and something very readable to say about the news of today.
"It's like I'm in jail!" says Aguerre, 53, grimacing. He is co-founder, with his brother Santiago, 51, of Reef, the sandal and surf fashion empire they sold in 2002 and 2005 to VF Corp. for about $100 million. After 20 years in the business, they took their top employees to Fiji for a week of surfing and at the end, at a dinner on a yacht, they said, "We're selling."
"Life is too short. Don't waste it. Go surfing."
This was Aguerre's favorite slogan at Reef. It branded the company — along with Brazilian girls in bikinis and surfers having fun, and palm trees swaying in a tropical breeze.
Branding is everything to Aguerre. Branding is what made the brothers rich.
And branding — the rebranding of America — is the way out of this country's long, dark financial debauchery, Aguerre believes, the escape from our decade adrift in the horse latitudes, our loser doldrums. We are, after all, the nation of Apple, Google, Coca-Cola, Jeep and Timberland.
"Why do you think VF bought Timberland for $2 billion? It's an American brand! They think there's way more money in an American brand. They're going to open a million stores. VF (the largest apparel company in the world) is $10 billion. Nike is $20 billion. And VF realized that it is action brands that are bringing equity and growing. Listen to the CEO's quarterly report. How are you going to grow Lee and Wrangler? There are no more cowboys. People don't want to be cowboys. People want to be Kelly Slater and Tony Hawk. PPR owns Gucci and Yves Saint Laurent. Yet last month they bought Volcom for $600 friggin' million. There is only one ideology left in the world, and that is consumption, and consumption is led by brands! I'm not saying I like unbridled capitalism because I've seen the downside. But if we lose the American dream, there will be nothing left for America to sell!"
Politics concern Aguerre not just because he is a political animal from a country (Argentina) where politics is a life-and-death game, but because a prosperous country is a happy country and happiness sells.
"Look," Aguerre lifts himself off the couch, "this is the best south swell in 11 years and I can't surf it!"
Surfing was there before the Aguerres, of course, and surf fashion, too, but the brothers brought a certain Latin DNA to it all.
"I looked at the American surf magazines. We couldn't afford radio or TV, and I thought what is this, all these guys in the same wet suits, similar bathing suits, shot by the same photographers? Last time I looked, surfers, most of them, liked girls, and there were no girls. … So we submitted a picture of a Reef girl, and the magazine refused to print it. I told the editor, OK, then I want a blank page, with the type: This magazine refuses to print what you can see in — I mentioned their main rival — and in 30 minutes, I get a call back. We had our first good ad."
At Reef in San Diego, everybody was allowed to come to work an hour late, provided (1) there was at least 4-foot surf and (2) they brought in a wet suit, wet. Business meetings ended up with a party at Fernando's house.
Some people say Fernando and Santiago still give the best parties in La Jolla. To be sure, the brothers' annual Liquid Nation Ball (Saturday, $300 a ticket) is not Las Patronas (charity group), ballroom dancing at the La Jolla Beach & Tennis Club. This is Flea of the Red Hot Chili Peppers wailing away on guitar as Fernando auctions off his own clothes to the highest bidder down to some crazy black-striped skivvies that maybe Greg Noll wore back in the day and high-fashion, low-clothing Argentine hat models dance with pro surfers like Rob Machado shaking his locks and Kelly Slater, probably not shaking his, while Che (the original Michael Korda negative) and Gandhi and Obama (body surfing) smile down.
All in the name of boobies.
Aguerre explains: "I was one day floating in the pool after my disc went wrong. I could have been dead. I could have had a heart attack … I could have been a cripple not able to walk. Something like this happens, and you realize how many things you need to do. How you have let some of your intense passions, your hopes, escape from you. 'Life is what happened when you were busy having other plans!' " he quotes John Lennon.
"I am now focused on Liquid Nation, raising as much money as I can for Shaney jo Darden and cancer education for young girls, (Aguerre has 15-year-old triplets of his own) the humanitarian award at the Liquid Nation Ball. She told me they have 8 million 'I love boobies' wrist bands now in their Keep A Breast campaign." He jumps into a laugh.
Aguerre hobbles to the door. "This rich guy comes and wants to buy my house for many millions of dollars," he says. "I look around. I check out other houses. But where am I going to move? I'm just a poor Latin immigrant!"
"Steve," he calls after me, "I'm just a serious guy who cracks a lot of jokes. You ever read Castaneda?"
I'm firing up my electric Dog Leg bike. "Yeah, back in the day."
"Death is the best adviser. When you're worrying about this and that, Death says, 'Nothing matters if I touch you!' Death is always around. Some stupid drunk driver hits you on that electric bike. A pilot takes a bad sleeping pill that didn't fade out in his system and poof, that's it for me."
Surf fashion — it's a lighthearted business, at the top.
Steve Chapple is an award-winning author of many books, including "Kayaking the Full Moon" and "Rock 'n' Roll Is Here to Pay." He has appeared on the Larry King and Charlie Rose shows, speaking on business and adventure. He writes for The New York Times and The New Yorker. Raised and residing in La Jolla and educated at Yale and the Scripps Institution of Oceanography, he lives above Windansea Beach with his wife and their three children. He can be reached at email@example.com © 2017, The San Diego Union-Tribune


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No Idiots Allowed Trailer (2015)
Documentary: Once a year the small and remote town of Neskaupstaður (Iceland) is invaded by hoards of metal fans who are attending Eistnaflug, Iceland's only heavy metal festival. Although small in scale the festival, now held for the 10th time, has played host to many of the worlds biggest metal bands.
Watch the official No Idiots Allowed 2015 trailer in HD below or find more No Idiots Allowed videos on Vidimovie.


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Limestone Service Station - Service Dept.
Review for Limestone Service Station - Service Dept.
Hector R. on 06/16/2015
Hector said the customer service at this business could have been more friendly.
Vehicle: Subaru Legacy
Category: Service
Service Date: 06/12/2015
Review Created: 06/16/2015
This review was collected via phone interview by Customer Research Inc.


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What if I told you that at least 90% of what you have read, watched or heard about supplements is much more about marketing hype and paying athletes exorbitant fees to endorse products rather than providing information that can truly help you get fit, healthy and strong? What if I told you much of the money or time you have spent in the past on high-priced supplements were pretty close to being a total waste? What if I told you there's a much simpler way to get the body of your dreams, with just a few of the tried and proven supplements, some smart training, the willingness to work hard and the right attitude? Would you be interested?
Introducing Supplement Secrets: An Inside Look at the Most Popular Training Supplements. This powerful tool will provide you with everything you need to finally achieve your dream of shedding the tummy fat for good and becoming the strongest, fittest you. Whatever your goal is, you can make promises to yourself all day and night, but actually sticking to your goals is the hard part! Within this guide you will discover everything you need to know to get started with proper supplementation and not be lead astray by the latest and greatest "miracle" supplement.
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How to transform your body with a few core supplements
Discover how to stack supplements to gain a synergistic effect quickly
Get results by avoiding these common pitfalls
What supplements you should never take
And much, much more
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About time someone exposed these scams!
- peggy
Yes, I was able to listen to this on an hour jog, and I never looked at a supplement the same way ever again!
They defining of whey and soy protein.
- Ntussia


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Most vehicle owners know that preventative maintenance is key for keeping your car in good working order. If you want to maximize fuel efficiency, ensure optimal operation and avoid unnecessary repairs, the best thing you can do is take your vehicle in for regular servicing. This includes regular oil changes. At Meineke #2377 in Puyallup, we recommend oil and oil filter changes at regular intervals. Consider it an investment in the lifespan of your vehicle. To learn more about oil change best practices, oil filter price comparisons and more, we invite you to contact us today.
Why do you need to replace engine oil or invest in replacement oil filters? Here's how oil works: It literally serves as a kind of car lube, keeping the pieces of the engine from rubbing against each other and causing too much friction. It also helps transfer heat away from the engine. It can prevent wear and tear while extending the lifespan of your vehicle significantly. This is why it is crucial to remove the old oil, which loses its efficacy, and replace it with new oil. Doing this on a regular basis will enable you to continue driving in streets of {city}, {State} safely. Visit in Puyallup today or make an appointment for an oil change.
A common question we get at Meineke #2377 is the question of how often an oil change is necessary. The short answer is that it just depends. There is no one right answer across the board; it simply depends on the make and model of your vehicle. What we can say without reservation is that it's much less often than it used to be. Oil and filter technologies have come a long way, so today's cars can go much greater distances without needing maintenance. To find out how often you should get your oil changed, consult with your owner's manual or bring your car to Meineke #2377 4021 S. Meridian for service.
What happens if you skip your regular auto filter change from Meineke #2377? The simple answer is that your vehicle will ultimately fall apart. The engine will eventually overheat and may cause the individual engine components to melt or to become warped. Additionally, the engine parts will create friction without the proper lubrication and will grind down much faster than they might have otherwise. The end result of all of this is that, sooner or later, your engine will simply stop working. You will be faced with the expensive prospect of a full-engine replacement, which is something far more demanding than the simple work of routine oil changes.
Not only is it a smart investment, but it can also be a highly affordable one. You can visit our oil change coupons page to see some potential ways to save money the next time you bring your vehicle to Meineke #2377 in Puyallup for an oil change!
Schedule an Oil Change today!Schedule Oil Change Appointment
4021 S. Meridian
Puyallup, WA 98373
Phone: (253) 200-1485
M-F 7:30 AM to 6:00 PM
Sat 7:30 AM to 6:00 PM
Sun 9:00 AM to 5:00 PM


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Healthy teams are more productive, and managers can support employee health by creating healthy workplaces. How does yours measure up? For each of the following items, assess your team and workplace on a scale of 1 (rarely or never), 2 (sometimes), or 3 (usually or always):
1. People come to work well-rested.
2. Employees are encouraged to recognize and share in each other's successes.
3. We find ways to make work more fun and interesting.
4. Most people want their work to have meaning and purpose.
5. There's a sense of community within the enterprise.
6. We take workplace safety seriously.
7. We have access to good healthy food (and not just junk) at work.
8. Team members are open to change and new ideas.
9. If one of our people needs help, the rest of us will offer it.
10. Senior managers take care of themselves and are in good health.
11. My people stay home if they're sick — and I'll send them home if they don't.
12. We laugh a lot at work.
13. Team members have chances to develop and use their talents.
14. My people remember to say hello to one another.
15. We resolve our differences before they become ongoing conflicts.
16. People get a decent amount of exercise, either on or off the job.
17. It's OK for people to take a little time at work to relax.
18. My team members have defined personal goals they're trying to achieve.
19. We respect and value diversity.
20. The enterprise has taken steps to promote employee wellness.
How well did you do?
Here's what your scores mean: If you scored below 30, you may have already noticed the effects of an unhealthy workplace. Look at ways to actively promote healthier lifestyles, reduce the stress levels, and eliminate specific health and safety risks (like the junk food in the vending machines, or unsafe conditions in the office).
If you scored between 30 and 50, you're in line with a lot of workplaces. But is your team really "average," or did the high and low scores cancel each other out? For example, your people may have healthy lifestyles and attitudes, but the workplace is stressful and competitive. Look for patterns in your responses.
If you scored above 50, congratulations! Your people are likely to stay healthy and productive, and they'll probably support and appreciate efforts to make the workplace even healthier.


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Incorporating G2DYM Aerials & Garex Electronics
Quality English products hand-crafted to order
AMATEUR - 15 November 2017
Proprietors;- Mr A.J. Nailer HNC BA G4CFY, Mrs J.R. Nailer.
Phone UK 01305 262250 International 0044 1305 262250
Please email or phone for a quote or to place an order by credit/debit card or for a Paypal money request. Purchase of several items is likely to make a saving on postage.
Orders & quote requests to email@example.com
Technical queries to email@example.com
BREAKING NEWS In the Editorial in November issue PW the Editor has stated that I am unable to submit articles for the forseeable future. This is not true! I am quite capable of submitting articles for Doing it By Design and for Technical for the Terrified BUT the Publisher for the new owner is unwilling to even discuss a requested small increase in my remuneration nor the shared copyright agreement and arrangement I had with the previous owners.
The Editor has further threatened that he will find other authors for my series and may not then be in a position to accept submissions from me. Considering the attitude taken by the Publisher and the Editor I will not be unhappy to retire as a PW author. I had not contemplated a retirement BUT having done this every month, with the exception of just two or three issues, since 2004, consuming on average in excess of 1/10 of my time, maybe it is time to stop.
| Book 1 includes the first 27 articles of
Technical for the Terrified, of which I am the author published
in PW between February 2005 and August 2009 and have been compiled into a 124 page book of the same name. The
writing style is aimed to be easy to read and should be of great value to readers of all technical levels. Subjects
include; a little maths, inductors & capacitors, DC circuits, transistor biasing, decibels, amplifiers, diodes &
rectification, the superhet, SSB & CW, FM & AM, bandwidth, Q, antennas & feeders, band-pass circuits,
RF/IF amplifiers, phase locked loops, HF transceivers, filtering in receivers, mixers, FET RF amplifiers.
Price is £12.00 plus £3.50 P&P.
Book 2 now available at £12 plus £3.50 P&P. Buy both books and pay only a single £3.50 P&P. Book 2 reproduces the next 23 articles from PW up to June 2013 and deals with components, equivalent circuits, crystal oscillators, amplifier stability, analogue low pass filters, transmitting ATU's, the superhet receiver, feeder cables and transmission lines, antenna gain and bandwidth, measuring RF power and SWR, measuring AC & DC voltage - inductance - capacitance - frequency & Q, circuit analysis, broadcast and television interference, singly balanced mixers, laying out PCB's, antenna advice for the listener, antenna tuning and the noise bridge, more on antennas and feeders, receiving 100kHz to 30MHz, homebrew receivers for the HF amateur bands, and circuit matching. Just so much in it.
Contents - In number and alphabetical order
|5W Broadband HF Linear
|Mixer - Osc for 4 metres
|Speech Processor
|7MHz DSB QRP TX
|Portland VFO
|7MHz DSB RX
|Poundbury 70MHz Front-end
|Toneburst, Piptone, & Kaytone
|4001 FM 4m Transceiver
|Poundbury SSB TX & RX IF
|VHF Masthead Preamps - Switched
|CTCSS module
|PSK31 Interface
|VHF Station Preamps - Switched
|HF Tuneable Preamp Switched
|QRP TX Attenuator
|VHF Transmit Amplifiers, 25/50/100W
4001 FM 4m TRANSCEIVER 70.2500 to 70.4875MHz in 20 channels in 12.5kHz steps. Originally AKD, then GAREX, now Spectrum. Really easy to use, not menu driven. Default channel at switch-on 70.450MHz. Now upgraded to give excellent audio quality on receive and transmit. RF output 5W or 25W presetable and switchable. Sensitivity better than 0.25uV for 20dB SINAD. Audio output 500mW rms. Supply requirement 13.5V DC 4A on high TX, 1.5A on low TX, 130mA on RX. This product is now being custom built to a high quality standard rather than down to the cheapest price. Type 4001S £195.00 UK, £203.00 EU, including tracked carriage & insurance.
2001 PIC module to change to 12.5kHz channel spacing and add CTCSS. Supplied with replacement reference crystal, LED in-between channel indicator and handbook with new frequency listing. £22.50 excluding P&P.
4001 replacement PIC to change to 12.5kHz channel spacing. Supplied with replacement reference crystal and handbook with new frequency listing. £8.50 excluding P&P.
6001 PIC module to change to 10kHz channel spacing and add CTCSS. Supplied with handbook with new frequency listing. £20 excluding P&P.
Upgrade kit for 2m, 4m, and 6m versions for improved IF amplifier, better RX filter matching and improved transmit audio. Supplied with replacement IF filters 10M07 and LTW455HT and complete circuit diagram & board component layout £14 excluding P&P.
7003 upgrade to receive IF amplifier and transmit modulator section is pending post design work but is likely to be similar to the kit above except that the 21.4MHz first IF filter will not be changed.
Noise Squelch, to give a quite background and excellent weak signal reception, upgrade kit £20.00 excluding P&P.
HF TUNEABLE PREAMP - SWITCHED, through handling power 100W. Fail safe operation, straight through path when switched off. 2-32MHz in 4 bands, with highly selective tuneable filters, reduces overload from strong broadcast stations. Gain 10dB at band edges and 25dB mid band, adjustable down to –20dB. 12/13.5V DC operation at 20mA. SO239 in/out connectors. Two-part stove enamelled box 150x100x75mm. Weight 410gms. Box kit £72.50 UK, £82.00 EU, £86.050 World. Ready built £123.00 UK, £129.00 EU, £133.00 World, including tracking & insurance.
|QRP TX ATTENUATOR, RF SWITCHED. This allows a rig running 5W output
to achieve QRP output levels of 500mW, 250mW, 100mW, and 50mW, using just three switches. Straight through path when
Box kit £49.50 UK, £56.50 EU, £60.50 World. Built £70.50 UK, £80.00 EU, £84.00 World. Including tracking and insurance.
|STATION PREAMPS RF Switched, for 2, or 4, or 6 metres. 100W through rated on transmit. RF & DC switched 0-20dB panel adjustable gain. 2m, 4m, & 6m types use BF964 with noise figure under 1.5dB. 10m type uses BFR84, with noise figure under 3dB. Over-voltage and reverse voltage protected. Protected against transients and EMP at input and output ports. PCB size 68 x 46 x 28mm. Purpose made box 76 x 70 x 38mm. Fitted with SO239 connectors. DC supply 13.5V 80mA. Types RP2S, RP4S, RP6S, & RP10S. PCB Kit £24.00 UK, £30.00 EU, £30.50 World. PCB & hardware kit £55.00 UK, £61.00 EU, £63.50 World. Ready built £79.50 UK, £87.00 EU, £89.75 World. Tracking & insurance included.
SWITCHED 100W through
rated on transmit for 2 or 4 or 6 metres. RF & DC switched and DC fed via the coax.
Gain 20dB, NF less than 1.5dB. Supplied with a DC/RF Station Box with SO239 connectors for the shack. Heavy duty waterproof
plastic box fitted with SO239 connector for feed coax and 1 metre of coax for connection to the antenna. Box size 82x80x50mm.
Types RP2SM, RP4SM, & RP6SM. PCB & hardware kit £57.50 UK,
£65.75 EU, £68.50 World. Ready Built £80.00 UK, £87.50 EU, £90.25 World, including tracking. Masthead fitting kit £8.00 extra.
MASTHEAD RECEIVE PREAMPSSWITCHED 400W through rated on transmit for 2 or 4 or 6 metres. RF switched. Supplied via separate wire DC feed. Gain 20dB, NF less than 1.5dB. Protected against reverse and over voltage and against EMC transients at input and output. Heavy duty waterproof box fitted with SO239 connector for feed line and 1 metre of coax to connect to antenna feed point. Box size 82 x 80 x 50mm. Types RP2SH, RP4SH, & RP6SH. PCB & hardware kit £49.25 UK, £55.25 EU, £57.75 World. Ready built £69.00 UK £76.50 EU, £79.25 World, including tracking. Masthead fitting kit £8.00 extra.
TRANSVERTERS. Early commercial transceivers often had a transverter facility with a low level output from the transmit driver stage and a means of disabling the power output stage. This meant there were two connections for an outboard transverter, one from the low power transmit port, the other for receive via the main antenna socket. Some of the top of the range rigs still have this facility but most modern rigs do not have any transverter facility at all. To use them with a transverter simply reduce the power to minimum and use a single connection to the transverter from the rig antenna socket. The RF sensing of the transverter will work at drive levels from 5W down to 100mW. Below that drive level, hard switching is required.
Add 2 or 4 or 6 metres to a rig with 10 metres, or 4 or 6 metres to a 2 metre rig. The transverter main board contains an attenuator in the transmit path and can be configured for 1-500mW or 0.5-5W drive levels and gives 300mW output. Class AB linear operation gives all mode use. The standard product includes a linear amplifier giving panel adjustable output 5 to 20W on 2m, or 5 to 25W on 4 or 6m. All spurious and harmonic outputs better than 45dB below carrier. Capable of withstanding long periods of transmit into bad SWR. Receive side has panel adjustable gain 0-20dB gain and NF less than 1.5dB. PCB size 150 x 93 x 18mm. Box size 200x160x77mm. Suffix s is used for single IF drive arrangement, suffix d is used for dual IF drive version, the suffix L is for integral linear.
|PCB Kit £69.50 UK, £76.50 EU, £77.50 World. PCB built £99.00 UK, £105.00 EU, £106.00 World. Box kit £110.50 UK, £119.25 EU, £125.25 World. Ready Built £173.00 UK, £179.00 EU, £185.00 World. Prices including tracking & insurance.
|PCB kits £173.00 UK, £177.50 EU, £180.25 World. PCB's built £213.50 UK, £218.25 EU, £221.00 World. Box kit £215.00 UK, £222.00 EU, £228.00 World. Ready built £283.50 UK £290.00 EU, £296.00 World. Prices include tracking & insurance.
Repeater shift for 2m output versions, add £8.50 to PCB kit or Box kit, £15 to PCB built or Boxed built.
ICOM COMPATIBLE TRANSVERTERS A version of transverter specially dedicated to certain ICOM rigs, complete with 60cm long cables, ready to plug-and-play. It is known to be suitable for the IC756Pro, 756Pro2, 756Pro3, 775, 781, 7600, & 7800, and any other rig not mentioned which requires just one TVTR IN/OUT RF lead plus a 7 pin DIN plug with a switching signal on pin 6. Types TRC2-10L/IC1, TRC4-10L/IC1, & TRC6-10L/IC1. Box kit £216.50 UK, £223.25 EU £229.25 World. Ready built £289.50 UK, £296.00 EU, £302.00 World.
NOTE: The following ICOM rigs are NOT plug and play compatible, but can be used with a standard transverter: 77, 78, 703, 706, 707, 718, 725, 726, 728, 729, 736, 737a, 738, 746, 746Pro, 756, 7000, 7400.
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TRANSMIT AMPLIFIER(20/25W) For use with Spectrum 0.5W Transverters. Two stage PTT switched class AB linear amplifiers for 2, or 4, or 6 metres giving a minimum of 20W out for the 2m version, and a maximum of 35W for the 6 metre version. Should not be run at more than 25W. Antenna relay with high speed TX/RX switching makes it suitable for all modes including data and RTTY and Packet. PCB size 125 x 57 x 28mm. Types TA2S4, TA4S4, TA6S4. PCB kit £106.00 UK, £112.25 EU, £113.25 World. PCB Built £121.00 UK, £124.50 EU, £125.50 World. Prices include tracking.
TRANSMIT AMPLIFIER(25W) Single stage switched class AB linear amplifier suitable for CW, SSB, FM, RTTY, and PACKET. 4m and 6m versions 1W in gives 8W and 5W in gives 30W output. 2m version 1W drive gives 6W out and 5W drive gives 25W out. Switching circuit senses RF & DC and uses a monostable relay driver giving VERY FAST attack and a hold time of 250ms to 2s for SSB use. For data modes the hold function can be disabled. PCB size 102x72x25mm. Diecast box size 171x120x55mm. Fitted with SO239 connectors. Types TA2SA, TA4SA, & TA6SA. Box kit £88.00 UK, £97.00 EU, £101.00 World. Ready built £112.00 UK, £120.00 EU, £124.00 World. Prices include tracking.
TRANSMIT AMPLIFIER(50W) Single stage switched class AB linear amplifier suitable for CW, SSB, FM, RTTY, and PACKET. 4m and 6m versions 5W in 30W out, 7.5W in 50W out. Switching circuit senses RF & DC and uses a monostable relay driver giving VERY FAST attack and a hold time of 250ms to 2s for SSB use. For data modes the hold function can be disabled. PCB size 102 x 72 x 55mm. Diecast box size 11x120x55mm with a heatsink on top 152x99x32mm. Fitted with SO239 connectors. Types TA2SB, TA4SB, & TA6SB. Box kit £102.50 Uk, £111.50 EU, £115.50 World. Ready built £127.00 UK, £133.00 EU, £137.00 World. Price include tracking.
TRANSMIT AMPLIFIER 70MHz (100W) Single stage parallel pair of 50W devices in class AB for all modes, 20 to 22W SSB input gives 100W peak output when run on a PSU with 13.5V at a load current of 16A. Rated for long periods on FM and Data at a continuous output of 50W. Also can be used with AM with an output carrier power set to 25W and peak power of 100W. RF sensed switching with fast attack and 500mS hang to avoid relay chatter when on SSB. Unit includes a peak reading power meter that remains in operation whilst the unit is connected to the power supply even though the amplifier is not in operation. Overall size 187x118x94mm. Ready built only £240.00 UK, £247.50 EU, £255.00 World, including tracked carriage.
TRANSMIT AMPLIFIER (25W) & PREAMP. Linear amplifier and receive preamplifier together in one case with independent switching. Amplifier details as TA-SA above, together with 20/26dB gain preamp. Types TA/RP2SA, TA/RP4SA, TA/RP6SA. Box kit £125.00 UK, £131.00 EU, £137.00 World. Ready built £159.00 UK, £164.75 EU, £170.75 World. Prices include tracking and insurance.
TRANSMIT AMPLIFIER(50W) & PREAMP . Linear amplifier and receive preamplifier together in one case with independent switching. Amplifier details as TA-SB above, together with 20/26dB gain preamp. Types TA/RP2SB, TA/RP4SB, TA/RP6SB. Box kit £142.50 UK, £148.50 EU, £154.50 World. Ready built £181.50 UK, £187.25 EU, £193.25 World. Prices include tracking and insurance.
TRANSMIT AMPLIFIER for 2, or 4, or 6metres.250mW to 500mW input, 20w to 35W output, with RF & DC switching. Class AB operation gives clean output with intermod products 35dB down suitable for all modes CW, SSB, FM, & DATA. Designed as self contained amplifier as an add-on for 0.5W transverters such as MEON. Boxed size 127x102x38mm. Types TA2S2, TA4S2, & TA6S2. To be redesigned when time permits.
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PORTLAND VFO, a rock stable FET VFO for 6.995 - 7.205MHz for a direct conversion for the extended 40metre band, or 7.900 - 8.400MHz for use as part of a mixer VFO system as local oscillator on 70.000 - 70.500MHz band, or 5.0-5.5MHz for 20m and 80m rigs with a 9MHz IF. Now includes upper band edge adjustment, and meets the requirement for the Intermediate Licence VFO project. The Buffer (2A), mounted on the rear of the box, delivers 1.6V p-p into 50 Ohm with second harmonic 40dB down. Kit of 2 PCB's and parts £18.25 UK, £18.25 EU, £19.25 World. Drilled Box and PCB kit with potentiometer and feedthroughs £35.00 UK, £39.50 EU, £40.50 World. Ready built £46.00 UK, £50.50 EU, £51.50 World. Prices include tracking. State required frequency when ordering.
MIXER - OSCILLATOR,a module for use in conjunction with the Portland VFO to produce a very stable local oscillator signal for a 4 metre receiver or transmitter with a 9MHz or 10.7MHz IF. Uses either a 51.4MHz crystal for a 10.7MHz IF, and a 53.1MHz crystal for a 9MHz IF. Comprises an overtone oscillator, a 4 quadrant multiplier mixer, a filter and buffer amplifier. PCB kit including choice of crystal £26.00 UK, £26.00 EU, £27.00 world. Ready built £36.50 UK, £40.50 EU, £41.00 world.
POUNDBURY (2) 9/10.7MHZ SSB Generator and Receive IF.A traditional SSB generator incorporating a speech processor. Carrier suppression typically in excess of 50dB. The board now connects directly to an external SSB filter which has a termination impedance of around 560 ohms and a few picofarads. Other termination impedances can be accommodated by the change of a single resistor. The circuit has provision for USB and LSB carrier crystals which are DC switched. Receive section uses Fet and Mosfet IF amplifiers and a singly balanced diode product detector and discrete audio derived AGC. Includes 1W audio amplifier. The crystal filter and carrier crystals are not included in the PCB kit. PCB Kit £70.25. PCB built £100.00. Optional extras, SSB filter £18.50. Carrier Crystals £3.00 each. Mic gain 4K7 log pot, Volume 100K log pot, £1.75 each. Signal meter £9.00. 8 ohm Loudspeaker £2.00. P&P on optional extras £3.50.
POUNDBURY 70MHz FRONT END.A receive preamp and mixer together with a transmit mixer and three amplifier stages to give 250mW output on 4 metres. Works in conjunction with the Poundbury Generator & Receive IF, the Portland VFO, and the Mixer-VFO board to give a complete 4metre transceiver. Can also work with the TA4S4 amplifier to give 25W output. PCB & Parts kit including 2 panel pots £50.00. PCB built £69.75
6-POLE CRYSTAL SSB FILTER. 6 crystal ladder filters for 9MHz and 10.7MHz. Bandwidth at -6dB 2300HZ . Bandwidth at -60dB 6000Hz. Shape factor 2.6:1. Insertion loss 2dB. Termination 560 Ohms and less than 5pF. PCB size 60 x 27 x 15mm. Kit of parts £17.00 UK, £19.00 EU, £20.00 World. PCB built £21.50 UK, £23.50 EU, £24.50 World. Including carriage.
7MHz DSBQRP TRANSMITTER. A 250mW double sideband transmitter comprising microphone amplifier, diode ring modulator driver and output stage. Works in conjunction with a Portland VFO and can be combined with the 7MHz DSB receiver to form a transceiver. The PCB kit of parts including a TO5 heatsink, a 10k log mic gain pot and wound toroids, price £33.00 UK, £37.00 EU, £37.50 World. PCB built plus log pot £41.00 UK, £45.00 EU, £45.50 World. Including carriage.
7MHz DSB RECEIVER. A double sideband direct conversion receiver requiring just the addition of a Portland VFO local oscillator. Works in conjunction with the 7MHz DSB QRP Transmitter and Portland VFO. PCB and kit of parts including volume control, £28.00 UK, £33.60 EU, £34.10 World. PCB built plus £40.00 UK, £43.65 EU, £44.15 World. Including carriage.
5W BROADBAND HF LINEAR AMPLIFIER Push-pull transistor HF linear covering the range 1.8-29.7MHz with full out for input levels between 37 and 97mW drive. Harmonics typically are 2nd -42dB, 3rd -18dB, 4th -49dB and 5th -29dB. This amplifier needs to be used together with a double Pi type low pass filter, details of which are supplied with the kit. Full kit of parts with drilled heatsink. Without wound toroids £36.00 UK, £41.65 EU, £44.15 World. With pre-wound toroids £43.00 UK, £48.65 EU, £51.15 World. Ready built and tested £51.00 UK, £56.65 EU, £59.15 World. Tracking included. Prices do NOT include the low pass filters.
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SPEECH PROCESSORto increase the sideband power and hence communications effectiveness of SSB and AM transmissions. The unit includes filtering which enhances the higher voice tones which carry most of the intelligibility. The peak to mean ratio of the transmission is improved by 10dB which has the same effect as increasing the transmit power by 10 times. It sounds nice too. Panel controls are included for clip level and output level. PCB size 82x45x17mm. Type SP1000E. Box kit £48.00 UK, £55.00 EU, £59.00 World. Ready Built £66.00 UK, £75.25 EU, £79.25 World. Prices include tracked carriage.
PSK31 INTERFACE, This is designed as an interface between a computer sound card and a transceiver. PCB kit £20.00 UK, £21.50 EU, £22.50 world. Box kit with cables and jack plugs £39.50 UK, £45.00 EU, £47.50 World. Ready built £54.00 UK, £59.50 EU, £62.00 World. Includes tracking.
CTCSS Module. Designed originally by Ken Ginn G8NDL, then modified and a new PCB layout by Spectrum. Produces all nine CTCSS tones that can be selected by a link or using a 9 way rotary switch. The unit is triggered by the microphone PTT and in turn provides a PTT output to the rig via an opto-isolator. The module includes a trimpot to set the tone level output to accommodate different microphone impedances. PCB size 67x55x10mm. PCB kit £18.00 UK, £19.50 EU, £20.50 World. PCB built & tested £25.00 UK, £27.00 EU, £28.00 World. Including P&P.
AUTO TONEBURSTA stable RC oscillator to provide a 500mS burst of 1750Hz audio for repeater access. The board has option for 7-10V or 10-16V operation. PCB size 29 x 27 x 15mm. Type AT1750. PCB Kit £7.00 UK, £9.00 EU, £10.00 World. PCB Built £10.00 UK £12.00 EU, £13.00 World. Including carriage.
PIPTONEend of message tone for 250mS for weak signal SSB use. The unit provides 50mV across a 600 ohm microphone. The tone and amplitude are presetable. A relay is included for receive and PTT switching. PCB size 44x25x17mm. Type PT1000S. PCB Kit £9.00 UK, £10.50 EU, £11.50 World. PCB Built £13.25 UK, £14.75 EU, £16.00 World. Prices include carriage.
KAYTONE(a name coined by Spectrum), this unit is the successor to the original Kaytone sold in 1979. End of message morse K particularly useful for weak signal situations. Adjustable pitch, speed, and level. A relay is included for TX and RX switching making it suitable for all rigs. PCB size 58 x 39 x 15mm. Type KT2000. PCB Kit £14.25 UK, £16.25 EU, £17.25 World. PCB Built £21.25 UK, £23.25 EU, £24.25 World. EU & World tracking £5.50 extra.
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Results 1 to 3 of 3
Thread: db2move <database> LOAD error
11-18-09, 12:45 #1Registered User
- Join Date
- Apr 2009
- Posts
- 23
Unanswered: db2move <database> LOAD error
I'm using DB2 V9.1 with Unix. Trying to migrate a particular database, copying data and lobs from version 8.2 to version 9.1. Here is what I did:
Export the data from source:
db2move <database> export
Which includes count, word count, and byte count:
ls -l | wc 99 884 6411
Export the lobs to target:
db2move <database> export l /db2_data/data/
ls -l | wc 99 884 6723
This includes an extra ddl and no system.msg file on target. That is why the count is the same. Not sure if the system.msg file matters or not.
Now I'm trying to load the data and lobs into the tables:
db2move <database> LOAD lo INSERT l /db2_data/data/
This is the error message I get:
Application code page not determined, using ANSI codepage 819
Error opening report file. Terminating ...
**Error occured while opening a file.
Could someone help me out or have any ideas what could be causing this error?
Thank you.
11-18-09, 13:02 #2Registered User
Provided Answers: 5
- Join Date
- Jan 2003
- Posts
- 4,310
I do not know the solution to your particular problem, but if you are migrating the entire database from the same OS that you are migrating to, why not just backup on 8.2 and restore to 9.1?
11-18-09, 13:03 #3:-)
Provided Answers: 1
- Join Date
- Jun 2003
- Location
- Toronto, Canada
- Posts
- 5,516
Check that db2move.lst is readable and not damaged.---
"It does not work" is not a valid problem statement.


text:
Opal tiara
Opal tiara
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Featuring delicate filigree metalwork that is hand-set with an intricate pattern of dazzling Swarovski crystals, this tiara's glam-factor is fit for a queen!
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Leslie Cheung (Cheung Kwok-wing), (born Sept. 12, 1956, Hong Kong—died April 1, 2003, Hong Kong), Hong Kong actor and singer who achieved enormous popularity in Hong Kong and throughout Asia first by means of his singing and then through his performances in Chinese-language films, in which he was one of the few actors willing to portray homosexual characters. He gained worldwide fame with his performance in the 1993 film Farewell, My Concubine, for which he was awarded the Grand Prix at the Cannes Film Festival.
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DBK has been awarded a renovation project for Victoria Park Apartment Community with BH Management, in North Florida. The project consist of: Seal Coat & Stripe, Installation of 520 Entry Lights, Construction of Dog Park, Resurface of Community Pool, Construction of 2 Outdoor Kitchens Wrapped with Stacked Stone, Granite Counter Tops and Stainless Steel Grills, Resurfacing of Tennis Courts, Laundry Area Renovation, Exterior Paint of 520 Units, Gutter, Stucco and Concrete Repairs, Installation of Drain Boxes and Drain Lines throughout Property!
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House Sitter in West Chester
I'm a self employed online college student who loves new pets and new places. I've been around cats, dogs and horses all my life, and I'll be sure to love your pets like I've loved my own. I don't party, smoke, or drink, and am very orderly when it comes to housekeeping.
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"The final professional word for the practitioner in family and child law" Phillip Taylor MBE...
Essential reading for all involved in the study and practice of family law
The papers of the 2011 Interdisciplinary Conference at Dartington Hall


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Hotel Features
Fitness Center
Fitness and travel go hand and hand at our Fitness Center. That's why we want to provide you with a state-of-the-art experience during your workout. Our hotel's Fitness Center is open 24 hours a day, seven days a week and features a wide range of weight machines and cardio equipment.
Enjoy the luxury of the Sheraton Club Lounge, an elegant setting in which to gather with friends or unwind with TV or free Internet. Club level guests enjoy access to the Club Lounge with complimentary breakfast and evening hors d'oeuvres.
Business Center
Our lobby business center features complimentary high-speed Internet access at PC workstations and remote printing capabilities. Or, stay connected on your own laptop with free wireless Internet. The business center is open 24 hours a day and seven days a week for your convenience.
With Starbucks nearby, it's easy to grab a bite to eat and refreshing drink and keep the conversation going with friends, family or colleagues.
Enjoy complimentary high speed Internet access in all of our guest rooms, business center and public areas.
Parking + Shuttle Service
Parking: Hotel guests enjoy complimentary self-parking. Long-term parking is available.
Shuttle Service: We offer complimentary shuttle to the surrounding area and based on availability. Please contact the hotel for schedules or for more information.
Make a Green Choice
Conserving water, energy and other resources is rewarding for you and great for the environment. Enjoy a $5 voucher at participating food and beverage outlets or 500 Starpoints® awarded at check-out for each night you decline housekeeping (except day of departure).
To participate in the Make a Green Choice program, please tell us at check-in or look for the doorhanger in your guestroom.
Delivery Services
Guests are invited to ship items to and from the hotel for a fee of $5.00 per item. Please inquire at the front desk with any questions.
Pet Friendly
Please note: There is a weight limit of 40 lbs. for dogs who visit the hotel. Our hotel allows up to one dog per room complimentary, each additional pet will be subject to a $25.00 fee.
A waiver must be signed at the time of check-in before the dog is allowed in any guest room or public area.
Fees: Please be advised that if excessive damage to the guest room is caused by the dog and repairs or deep cleaning is required, the guest will be liable for those costs. If damage is discovered after the guest departs, the credit card on the guest record will be billed for these additional costs.
Limitations: Dogs are not permitted in the restaurant, pool, or in the club lounge during breakfast or evening reception hours) and may not be left unattended in the guest room at any time during the guest's stay.
Sheraton St. Paul Woodbury reserves the right to move or change room type of reservations with a dog.


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Project Description
A high-energy, fast-paced student speaking event.
Similar to TEDxUSU, 9 student researchers share the passions, experiences and stories behind their research. The twist is that each speaker has only 5 minutes to talk and their 20 slides automatically advance.
Join us after the event in the main atrium of the Merrill-Cazier Library for food, music and a chance to mingle with the speakers.
April 13, 2018
12 – 1 p.m. | Reception to Follow
Merrill-Cazier Library
Open Invitation
Casual Attire