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Effros, R. M., C. Darin, E. R. Jacobs, R. A. Rogers, G. Krenz, and E. E. Schneeberger. Water transport and the distribution of aquaporin-1 in pulmonary air spaces.J. Appl. Physiol. 83(3): 1002–1016, 1997.—Recent evidence suggests that water transport between the pulmonary vasculature and air spaces can be inhibited by HgCl2, an agent that inhibits water channels (aquaporin-1 and -5) of cell membranes. In the present study of isolated rat lungs, clearances of labeled (3HOH) and unlabeled water were compared after instillation of hypotonic or hypertonic solutions into the air spaces or injection of a hypotonic bolus into the pulmonary artery. The clearance of 3HOH between the air spaces and perfusate after intratracheal instillation and from the vasculature to the tissues after pulmonary arterial injections was invariably greater than that of unlabeled water, indicating that osmotically driven transport of water is limited by permeability of the tissue barriers rather than the rate of perfusion. Exposure to 0.5 mM HgCl2 in the perfusate and air-space solution reduced the product of the filtration coefficient and surface area (P f S) of water from the air spaces to the perfusate by 28% after instillation of water into the trachea. In contrast, perfusion of 0.5 mM HgCl2 in air-filled lungs reducedP f Sof the endothelium by 86% after injections into the pulmonary artery, suggesting that much of the action of this inhibitor is on the endothelial surfaces. Confocal laser scanning microscopy demonstrated that aquaporin-1 is on mouse pulmonary endothelium. No aquaporin-1 was found on alveolar type I cells with immunogold transmission electron microscopy, but small amounts were present on some type II cells.
- 28-kDa channel-forming integral membrane protein
- mercuric chloride
- epithelium
- endothelium
- immunogold
functional evidence for water channels in cell membranes was first reported by Paganelli and Solomon (24) and Sidel and Solomon (32) nearly 40 years ago. They found that osmotic water permeability exceeds diffusional water permeability in red blood cells and attributed this difference to the movement of water through pores in the red blood cell membranes. Macey (19) subsequently showed that water transport across red blood cell membranes is inhibited by organic mercurials and that the activation energy of water movement through red blood cell membranes is low in the absence of mercurials but high when they are present. This suggests that in the presence of osmotic gradients, most of the water moves through aqueous channels that can be inhibited by mercurials. The structure of the red blood cell water channels has been recently elucidated by Agre et al. (1), who cloned a protein of 28,000-Da molecular mass, designated as the channel-forming integral membrane protein (CHIP28). This represented the first member of a growing family of membrane proteins referred to as "aquaporins," many of which transport water, and CHIP28 is now referred to as aquaporin-1.
When lungs are perfused with hypertonic solutions of NaCl, urea, or sucrose, the fluid that is extracted from the pulmonary parenchyma is essentially free of any solutes (7). This observation was consistent with the hypothesis that most of the water removed from the lungs with these perfusion solutions crosses cell membranes, which are impermeable to extracellular electrolytes and other small hydrophilic solutes. In a subsequent study (11), our laboratory found that perfusion of fluid-filled lungs with hypertonic solutions of glucose or sucrose resulted in the loss of virtually solute-free water from the air spaces, suggesting that most of the water had crossed cellular membranes rather than through the intercellular junctions.
Nielsen et al. (23) recently reported that the mRNA for aquaporin-1 is present in the endothelium of lungs and a variety of other organs with continuous endothelial membranes. Hasegawa et al. (15) found that mRNA encoding aquaporin-1 is present in a subpopulation of pulmonary alveolar epithelial cells, tracheal epithelium, and the tracheal adventitia. Immunohistochemically stained lung tissues examined by light microscopy revealed what was interpreted as strong staining of the pulmonary epithelium. Folkesson et al. (12) showed that the osmotic water permeability of freshly isolated rat alveolar type II cells is weakly temperature dependent and inhibited by 0.5 mM HgCl2. Because it has not been possible to isolate type I alveolar cells, which cover 97% of the alveolar surface, these investigators conducted additional experiments in perfused sheep lungs. Instillation of hyperosmolar saline (900 mM NaCl) into these lungs resulted in the flow of fluid from the vasculature into the air spaces. This movement of water was inhibited by HgCl2 in a manner suggesting that aquaporin-1 molecules play an important role in the movement of water between the air spaces and vasculature of the lungs, and they suggested that much of the aquaporin-1 is located on the pulmonary epithelium. However, it was not determined whether the osmotic shift of water across the alveolar-capillary barrier was limited by permeability of the membranes rather than the rate at which the lungs were perfused. In view of the observation that HgCl2 can increase pulmonary vascular resistance (2), the effects of HgCl2 might reflect a decrease in the alveolar-capillary surface area of the lungs rather than inhibition of aquaporins in the alveolar membranes. Furthermore, they did not determine whether endothelial rather than epithelial aquaporin-1 activity might be responsible for the effect of HgCl2 on water transport across the alveolar-capillary barrier.
In the present study, the movement of water in response to osmotic gradients was monitored after intratracheal instillation and pulmonary arterial injections by using both labeled (3HOH) and unlabeled water (HOH). New indicator dilution approaches were used to estimate the osmolality of the pulmonary vasculature after instillation of hypotonic and hypertonic solutions into the air spaces and to determine the effect of HgCl2 on epithelial and endothelial permeability to water. In addition, light- and electron-microscopic immunochemical studies were used to localize more precisely the distribution of aquaporin-1 in the lungs.
Water and Solute Exchange
Experimental procedures.
The physiological studies were divided into two sets of experiments (Table 1). In the first set (groups 1–7), 5 ml of hypotonic or hypertonic fluids containing 3HOH were instilled into the air spaces of isolated, perfused rat lungs, and measurements were made of the movements of labeled and unlabeled water between the air-space and vascular compartments. In the second set (groups 8 and 9), 0.7 ml of solute-free water with 3HOH were injected into the pulmonary artery of air-filled lungs, and samples of the pulmonary venous outflow were collected to determine how much of the 3HOH and HOH had exchanged between the tissues and vasculature. No attempt was made to subdivide the cellular and interstitial compartments of the lungs in these studies.
In all studies, Sprague-Dawley rats [average wt 343 ± 67 (SD) g, n = 46] were anesthetized with 0.7 ml of a 64.8 mg/ml pentobarbital sodium solution. The chest was opened, catheters were placed in the trachea, pulmonary artery, and left atrium, and blood was flushed from the vasculature at constant temperature (see Table 1) with 10 ml of perfusion fluid that contained (in mM) 25 NaHCO3, 110 NaCl, 4 KCl, 2.5 CaCl2, and 0.8 MgSO4, as well as 150 mg/dl glucose, 10 mg/dl urea, and 5 g/dl bovine serum albumin (BSA; Cohn Fraction V, 98–99% albumin, Sigma Chemical, St. Louis, MO) adjusted to pH 7.4 with 1N NaOH when exposed to 5% CO2. The albumin was labeled with Evans blue (EB; 10 mg/l). The lungs were ventilated with a 5% CO2-95% O2 mixture at a tidal volume of 4.5 ml and at a rate of 30 breaths/min. Ventilation was discontinued just before the beginning of the experiments. Pulmonary arterial pressures were measured just proximal to the pulmonary arterial catheter.
In the first set of experiments (groups 1–7, Table 1), perfusate was pumped through the lungs at either 6.45 or 22.1 ml/min with a nonpulsatile pump from a reservoir that was heated to 37°C, cooled to 11°C, or left at room temperature (25°C) to keep the pulmonary arterial temperature constant. Temperatures were measured with a thermocouple inserted into the arterial line just proximal to the point at which the catheter entered the pulmonary artery. Left atrial pressures were kept at zero. After 70 s of perfusion, an experimental solution was instilled into the air spaces with a no. 50 polyethylene catheter that was inserted through the endotracheal tube to the carina. The outflow from the left atrium was collected in serial beakers over a 120- to 240-s interval, and, in some experiments, samples were collected from the air spaces at the end of the experiments.
In the second set of experiments (groups 8 and9, Table 1), 0.7 ml of water with3HOH, [14C]dextran, and 1 g/l albumin labeled with 10 mg/l EB were introduced into the pulmonary arterial line (see description of perfusate above), and the outflow from the left atrium was collected at 2.66-s intervals with an automated collection rack over an 80-s interval. These injections were made with a chromatography valve, which enabled us to rapidly switch the inflow from tubing that was filled with the control perfusion solution to tubing that contained the injection bolus.
Indicator and solute concentrations were measured in 0.4-ml samples of the injected solutions, the perfusate, and, in group 3 and 4 experiments, the fluid remaining in the air spaces at the end of the experiments. EB-labeled albumin concentrations were measured in 60-μl samples that were diluted 10-fold in isotonic saline and then centrifuged at 1,000g for 10 min. The supernatant was removed, and the absorbance was determined spectrophotometrically at 620 nm. Concentrations of3HOH and in the second group of experiments, [14C]dextran, molecular weight 45,000, were determined with samples of perfusate and the instilled solutions by diluting 50 μl in 2 ml of Hi-Safe III scintillation fluid and counting these samples with an automated beta counter. Corrections were made for background radiation and crossover between the 14C and3H labels. Na+([Na+]) and K+ concentrations ([K+]) were determined with ion-specific electrodes. In most of the group 1–7 studies osmolality was measured with a vapor pressure osmometer. In the group 8 and9 studies (see Table 1), osmolality was estimated from EB-labeled albumin concentrations. Previous studies from our laboratory (7) indicated that solvent drag reflection coefficients (ςf) of small hydrophilic solutes and electrolytes across the endothelium are close to 1.0. After introduction of water into the arterial inflow, the loss of water from the capillaries should result in increases in osmolality that are proportionate to those of EB-labeled albumin Equation 1where cv and ca are the venous and arterial osmolalities, and [EB]v and [EB]a are the concentrations of EB-labeled albumin in the venous and arterial perfusate. When air-space samples were collected, [Na+], [K+], and osmolality were measured (none of the EB entered the air space). The conditions of each set of experiments are indicated in Table 1. Binding of EB to albumin in the experimental solutions was checked by filtering them through ultrafilters (Centricon) with a molecular cutoff of 30,000. No unbound albumin was detected in these experiments. The binding of EB to albumin and its color remained uninfluenced between osmolalities of 10 and 500 mosmol/kgH2O. Measurements of lactic dehydrogenase (kit, Sigma Chemical) were made in the air-space and perfusate solutions at the end of some of the experiments, but none was detected.
Calculation of water transport.
After instillation of hypotonic solutions into the air spaces (groups 1–3 and5–7, Table1), entry of water into the pulmonary exchange vessels increased the flow leaving these vessels and decreased EB-labeled albumin concentrations. Assuming no loss of EB and steady-state conditions, the venous outflow (Fv) was calculated from the arterial inflow (Fa), [EB]a, and [EB]v with the equation Equation 2 The clearance of water (Choh) from the air spaces to the perfusate was calculated from the difference between the venous and arterial flows Equation 3 The volume of water remaining in the air spaces at time i(Vas,i) was assumed to be equal to the amount originally instilled into the lungs (5 ml) minus the volume of water that was transferred from the air spaces to the perfusate before that time Equation 4where Δti represents the duration of the time interval during which theith sample was collected.
The rate of loss ( ) of3HOH from the air spaces during the ith interval was calculated with the equation Equation 5where [3HOH]v, iis the venous concentration of3HOH in blood collected from the left atrium during the ith interval.
The amount of 3HOH remaining in the air spaces during the collection of theith sample (3HOHas, i) was assumed equal to the original amount instilled into the air spaces (3HOHas,0) minus the quantity that had left in the venous outflow Equation 6 The concentration of 3HOH in the air-space fluid at the time that theith sample of perfusate was collected ([3HOH ]as, i) was calculated from Eqs. 3 and 5 Equation 7 The clearance of 3HOH (C3 hoh) from the air spaces during each collection interval was calculated with the equation Equation 8In the group 4 experiments, instillation of hypertonic saline into the air space resulted in the movement of fluid from the vasculature to the air spaces. This caused Fv to fall relative to Fa and increased the concentrations of EB-labeled albumin in the perfusate leaving the pulmonary exchange vessels. The clearance of water into the air spaces was calculated from Eq. 2 . This value was multiplied by −1 to yield a positive value for this parameter. In this group of experiments,3HOH was incorporated in the perfusate rather than the air-space solution, and it moved into the hypertonic fluid that had been instilled into the air spaces. The influx of 3HOH into the air space (Ias, i) was calculated with the equation Equation 9 C3 hohinto the air spaces was calculated on the basis of the concentration of3HOH entering the arterial end of the exchange vessels ([3HOH]a) Equation 10 For purposes of comparison, values were also calculated for the peak values of Choh divided by peak values of C3 hoh (see Table 2).
In the second group of experiments, the lungs were perfused with the EB-labeled albumin solution described above for 2 min, and a 0.7-ml bolus of water was injected into the arterial inflow. The injectate bolus contained 3HOH and [14C]dextran that were dissolved in water containing the same concentration of EB as that present in the perfusion solution and 0.1 g/dl albumin. As the hypotonic injection bolus traverses the pulmonary capillaries, HOH is lost from the vessels, resulting in an increase in the concentration of EB-labeled albumin. The rate at which HOH was lost from the vascular compartment to the tissues (which is equivalent to its clearance, C) was calculated by using the equation Equation 11The fraction (Dhoh) of the injected volume of water (0.7 ml) that was lost each second from the pulmonary venous outflow was calculated by dividing Choh by 0.7 ml (see Fig. 6). The fraction of the injected 3HOH (D3 hoh) that was lost each second from the perfusate samples was calculated in the following fashion. It was assumed that the labeled dextran (which was injected into the pulmonary artery with 0.7 ml of water containing3HOH) does not leave the vasculature during transit through the lungs. The concentration of3HOH (designated as the calculated value, [3HOH]a) that would have been present in each of the venous samples had there been no diffusion of 3HOH out of that sample in transit through the lungs was determined with the equation Equation 12where [dex]v represents the observed concentration of14C-dextran in the venous sample, and [dex]injdesignates the corresponding concentration in the injected bolus. D3 hoh of the injected3HOH that was lost from each sample because of diffusion into the tissues is therefore Equation 13
where Δt is the collection interval. The hypotonic bolus was followed by isotonic perfusate containing no 3HOH, and there was consequently return of both 3HOH and HOH to the vasculature at later times, shown as negative values (see Fig. 6). Presumably because of differences in the perfused volumes of different lungs, arrival of indicators in the outflow varied between experiments. To facilitate graphic presentation of these data, the samples with peak 3HOH are shown at "zero time" and other samples at the number of seconds before and after the peak 3HOH concentrations had been attained.
Calculation of the product of the filtration coefficient (P f) and surface area (S) of the alveolar-capillary wall [(P f S) ] was made with the following equation, which is derived in the Equation 14where vw is the molar volume of water (18 cm3/mol), cc represents the estimated osmolalities of the perfusate in the exchange vessels and Vinj is volume injected into pulonary artery (0.7 ml) (see ). Although the subscript c is used, it must be emphasized that exchange may not be limited to the capillaries but may include other microvessels within the lungs. The osmolality of the air-space fluid (cas) was zero ingroups 1,2, and5. In the other groups, cas was calculated from the amount placed in the air spaces, and subsequent changes in volume were calculated by using Eq. 3 . Values ofP f Sare expressed in units of milliliters per second, and mean values shown are based on the values calculated from samples collected near the peak of the clearance curves (60 s after instillation of fluid into the air spaces in groups 1 and3–7 and 30 s after instillation of fluid in group 2).
Similarly, theP f Sproduct of the endothelium [(P f S)endo] was estimated by using the equation Equation 15where cti, ndesignates the estimated osmolality of the tissue just before thenth sample had traversed the exchange vessels.
Statistical analysis.
Values are means ± SE. The significance of differences between mean peak clearance values were tested with a one-way analysis of variance. A Newman-Keuls test was used to compare individual mean values (Sigmaplot, Jandel, San Rafael, CA). A similar approach was used to compareP f Svalues, but a separate Student'st-test was also used to compare (P f S) in the presence or absence of HgCl2. Comparisons between peak losses of HOH and 3HOH in the pulmonary arterial injection experiments were made with a Student'st-test.
Morphological Studies
Rabbit anti-aquaporin-1 and preimmune sera were a generous gift of Dr. Dennis Brown (Renal Unit, Massachusetts General Hospital). Previous studies established that there was no difference in the specificity between the whole anti-serum and the affinity-purified anti-aquaporin-1 antibody, as determined by Western blot analysis and immunogold staining of rat kidneys (29). CY3-conjugated goat anti-rabbit immunoglobulin G was from Jackson Immuno-Research (West Grove, PA). Protein A gold conjugate (10 nM) was from Ted Pella (Redding, CA). Glutaraldehyde, acrolein, and Epon were obtained from Ladd Research Industries (Burlington, VT); uranyl acetate and propylene oxide were from Polysciences (Warrington, PA).
Female BALB/c mice, 6–8 wk old, were anesthetized with pentobarbital sodium (5 mg/100 g body wt). The thorax was opened, and fixative containing 2% paraformaldehyde, 75 mM lysine, and 10 mM sodium periodate (20, 21) was used to gently inflate the lung. After 5 min, slices of lung were fixed overnight in sodium periodate at 4°C. The next day, small pieces (1 mm3) of lung were cut, rinsed in phosphate-buffered saline (PBS), dehydrated in graded ethanol, embedded in Epon, and polymerized overnight at 60°C. Sections, 1 μm thick, were cut and dried onto Superfrost/Plus microscope slides at 60°C overnight. Before immunostaining, the resin was etched by treating the sections for 5 min with a solution consisting of 2 g KOH, 5 ml propylene oxide, and 10 ml methanol (20). After being washed in methanol-PBS (50:50 vol/vol) and PBS for 5 min each, sections were blocked for 15 min with 1% BSA in PBS. They were then incubated with either aquaporin-1 anti-serum or preimmune serum (both at 1:800) for 2 h, followed by goat anti-rabbit immunoglobulin G-CY3 conjugate for 1 h. All incubations were followed by two 5-min washes each of 2.7% NaCl and PBS. Sections were mounted in 50% glycerol in 0.2 M tris(hydroxymethyl)aminomethane ⋅ HCl, pH 8.0, containing 2% n-propyl gallate to retard quenching of the fluorescence signal. Sections were examined by using a Sarastro 2000 confocal laser scanning microscope (Molecular Dynamics, Sunnyvale, CA) fitted with a 25-mW argon-ion laser and adjusted for simultaneous transmitted light (lung tissue) and >535-nm fluorescent light (aquaporin-1) imaging. Transmitted light and fluorescent images of the lung and aquaporin-1, respectively, were superimposed by image processing.
Rats were anesthetized as described above, the thorax was opened, and 4% paraformaldehyde was gently instilled into the lungs. The lungs were removed, and sections were placed overnight in 2% paraformaldehyde and 1% acrolein at 4°C. For immunogold staining, ultrathin frozen sections of lung were prepared as described by Tokuyasa (36). Briefly, small blocks of fixed lung were placed in 10% gelatin in PBS, chilled, infiltrated with 2.3 M sucrose overnight and frozen in liquid nitrogen. Ultrathin cryosections, 60 nm thick, were cut on a Reichert FC4E ultracryomicrotome and mounted on Parlodion/carbon-coated Ni grids. They were stored overnight on drops of 2.3 M sucrose on solidified 2% gelatin. After being rinsed in PBS, sections were quenched with 0.02 M glycine-PBS and rinsed in PBS, each for two 5-min periods. Sections were preincubated for 15 min in 0.1% BSA-0.4% gelatin-PBS (alb/gel/PBS) for 15 min and then incubated with aquaporin-1 anti-serum and diluted 1:1,200 with alb/gel/PBS for 2 h at room temperature. After being washed in 0.1% BSA-PBS-0.3% Tween-20 (rinse solution), sections were incubated with protein A-gold (10 nm, 1:100) for 1 h and washed 3 × 5 min in rinse solution and once in PBS. Sections were then fixed for 10 min with 1% glutaraldehyde in PBS, washed in PBS, and rinsed 4 × 1 min in water. They were stained with 2% aqueous uranyl acetate for 4 min and exposed to 0.5% methyl cellulose in water for 16 s. Grids were dried and viewed in a Philips 301 electron microscope.
Water and Solute Exchange
Clearance of 3HOH and HOH from airways.
After instillation of 5 ml of solute-free water, 77 mM NaCl, or 308 mM NaCl into the air spaces, the clearance of HOH from the air spaces to the perfusate was invariably and significantly less than that of3HOH (see Fig.1 and Table 2). (P f S) in the control experiments averaged 5.77 ± 0.85 (SE) cm3/s.
Increases in the perfusion rate from 6.45 to 22.1 ml/min increased pulmonary arterial pressures (Fig. 2) and the rates at which both 3HOH and HOH were cleared from the lungs (Table 2). Of the 5 ml of water that had been instilled into the lungs, 1.74 ± 0.30 ml had been recovered in the venous outflow after 4 min of perfusion at 6.45 ml/min. In contrast, only 2 min were required to recover 2.35 ± 0.28 ml of the instilled water when the lungs were perfused at 22.1 ml/min. Increasing flow by a factor of 3.4 (from 6.45 to 22.1 ml/min) was associated with a twofold increase in (P f S)A(see Fig. 3).
The calculated value of (P f S) was increased when 77 mM NaCl rather than water was instilled into the lungs (Fig. 3, bar 3 vs.1). (P f S)Awas not significantly greater when hypertonic saline was instilled into the lungs rather than water (Fig. 3, bar 4 vs. 1). Cooling of the perfusate and airway instillate to 11°C slightly reduced mean values of (P f S)Afrom control values, but this effect was not significant (Fig. 3,bar 5 vs.1).
Incorporation of 0.5 mM HgCl2 in the air-space and perfusate fluids decreased mean values of (P f S)Aby 28% (Fig. 3, bar 7 vs.6). This effect was significant when the two groups were compared by using a pairedt-test (P < 0.05), but it was not significant with use of a Newman-Keuls test, and the effect that HgCl2 had on endothelial filtration was much more pronounced (Fig. 3, bar 9 vs. 8; see (P f S)endo and inhibition with HgCl2).
Electrolyte concentrations in the perfusate.
The inflow of water into the perfusate resulted in similar decreases in EB-labeled albumin concentration, osmolality, and [Na+] in the perfusate at slow flow (Fig.4 A). However, the relative concentrations of K+ were greater than those of the other indicators between 100 and 250 s after the water had been instilled into the lungs. This was particularly evident at slower flows (compare Fig. 4, A andB). [Na+] values appeared to be slightly lower than expected from the osmolality measurements at high flows (Fig. 4 B), but this could reflect limits of the precision with which these parameters were measured. Unlike the experiments in which water was instilled into the air spaces at 37°C, no evidence was observed for release of K+ from the lungs into the perfusate after instillation of 77 mM NaCl at 37 or 25°C, 308 mM NaCl at 37°C, or water at 11°C.
Air-space solute concentrations (groups 3 and 4).
Because of the rapidity with which water was lost from the lungs when there was no solute in the airway solution (groups 1, 2, and5), it was not possible to sample the airway fluid at the end of these experiments. However, samples of the air space could be collected after 240 s had elapsed when either 77 or 308 mM NaCl was instilled into the lungs (Fig.5). When 77 mM NaCl was instilled into the air spaces, both [Na+] and osmolality in the air space increased to values approaching those in the perfusate. Similarly, air-space [Na+] and osmolality rapidly fell after instillation of 308 mM NaCl into the air spaces and approached those in the plasma. No K+ was present in the solutions instilled into the lungs, but there was movement of K+ from the lung to the perfusate during the course of the experiments. Much more K+ entered the air spaces when hypotonic rather than hypertonic saline was instilled into the air spaces.
(PfS)endoand inhibition with HgCl2.
As described in methods, studies of endothelial permeability were conducted by injecting 0.7 ml of water into the pulmonary artery with3HOH and dextran. The lungs were perfused with EB-labeled albumin in these experiments, and the injection solution also contained this indicator. Initially, the EB concentrations increased as water entered the tissues of the lungs. EB concentrations subsequently fell below baseline levels after the hypotonic fluid had left the pulmonary microvasculature and water returned to the perfusate (Fig. 6). Once again, the movement of 3HOH out of the vasculature was significantly more rapid than that of HOH.
Perfusion of the lungs for 2 min with 0.5 mM HgCl2 decreased the loss of HOH from the perfusate without having a significant effect on peak losses of 3HOH, and ratio of peak Dhoh to peak Dtho fell from 0.68 ± 0.02 to 0.34 ± 0.02 (P < 0.01, Fig. 6, A andB). (P f S)endoafter injections of virtually solute-free water averaged 281 ± 61 ml/s and fell to 39 ± 3 ml/s when 0.5 mM HgCl2 was incorporated in the perfusate. Perfusion with HgCl2did not significantly affect pulmonary arterial pressures in these brief experiments, but increases in pressure were observed in more prolonged studies (not shown).
Morphological Studies
Confocal microscopy.
Half-micrometer-thick optical sections of aquaporin-1-stained lung were recorded with a ×60 objective (numeric aperture 1.4) present. Staining for aquaporin-1 was observed on the endothelium of all segments of the pulmonary capillary bed (Fig.7). At this level of magnification, however, staining of alveolar type I (ATI) and alveolar type II (ATII) cells could not be resolved.
Immunogold labeling.
At the ultrastructural level, aquaporin-1 was localized by immunogold labeling on the apical and basolateral surface of endothelial cells, as well as on the membrane of a number of endothelial vesicles (Fig.8). Aquaporin-1 was not detected in ATI cells, and immunogold labeling for aquaporin-1 in ATII cells was quite variable. When present, label was detected on both apical and basolateral surfaces of ATII cells. However, in comparison to endothelial cells, the degree of labeling was less, and some ATII cells in the same section remained unlabeled. Sections incubated with preimmune rabbit serum followed by protein A gold were consistently negative.
The high incidence and serious consequences associated with edema formation in the lungs have been responsible for continuing interest in the manner in which water moves between the pulmonary compartments. Three basic strategies have been used to study water exchange in the lungs. 1) Injections of labeled water (3HOH or2H2O) have been made into the pulmonary artery or air spaces, and its exchange with tissues has been measured.2) Hypertonic or hypotonic solutions have been injected into the pulmonary artery or air spaces, and the movement of HOH has been monitored.3) The movement of fluid out of the vasculature has been studied after increases in hydrostatic pressure. In this study, a direct comparison was made between tracer and osmotic water fluxes in the lungs.
Loss of Labeled Water From Air Spaces Is Limited by Perfusion
Because labeled water and most gases and lipophilic solutes rapidly cross cell membranes, it is generally assumed that at attainable cardiac outputs, concentrations or partial pressures of these substances are virtually the same in the air spaces and the plasma leaving the pulmonary exchange vessels. For example, similarities in the manner in which deuterated water (2HOH),3HOH, [14C] butanol, and14CO2emerge from the lungs in the pulmonary venous outflow after they have been instilled into the lungs (4, 9, 10) suggest that equilibration between the alveolar compartment and blood leaving the exchange vessels is complete for each of these indicators. Perhaps the most convincing evidence that 3HOH transport is limited by flow rather than permeability was provided by experiments in which the rates of transport of3HOH and14CO2from the air spaces to the perfusate were directly compared (10): when mixtures of 3HOH, H14 , and14CO2are instilled with carbonic anhydrase into the airways, the quantity of14C appearing in the outflow remains the same as that of 3HOH between a pH of 7.4 and 8.0. The fraction of the14C label in the form of the more diffusible species (14CO2) is 5% at a pH of 7.4 and 1.25% at pH 8 so that the gradient for diffusion is reduced by 75% at the more alkaline pH. Because the rate of diffusion of14CO2relative to that of 3HOH is uninfluenced by this change in the concentration gradient between the air spaces and perfusate, losses from the air spaces of both3HOH and14CO2must be limited by flow rather than by diffusion.
When indicator losses from the air spaces are limited by the rate of tissue perfusion, the permeability of the alveolar-capillary membranes cannot be measured. It is for this reason that the recent observation of Schnitzer and Oh (31) that uptake of3HOH by perfused rat lungs was limited by perfusing them with HgCl2 may reflect decreased tissue perfusion rather than inhibition of water channels. Diffusion of3HOH through cell membranes occurs primarily through the cell lipids, which are presumably not inhibited by HgCl2. Because movement across cell membranes by labeled water is very rapid, exchange with the tissues is limited by flow rather than the filtration coefficient of the endothelium. In a recent study by Carter et al. (2), diffusion of2HOH into the air spaces of mouse lungs was monitored with an intra-alveolar fluorescence technique. They reported that increasing flow rates from 1.7 to 2 ml/min accelerated movement of 2HOH into the air spaces, but a further increase to 2.3 ml/min was without effect, and they concluded that exchange was diffusion rather than flow limited at these higher flows. However, increases in flow in the latter experiments may have been accomplished by vascular recruitment rather than acceleration of the velocity of flow in the monitored vessels.
Osmotic Flow of Water Across the Alveolar-Capillary Barrier Is Limited by Permeability
The clearance of HOH among the air-space, tissue, and vascular compartments of the lungs in the present experiments was invariably slower than that of 3HOH, indicating that equilibration of the unlabeled species is limited by the permeability of the membranes separating these volumes. This observation might appear paradoxical because1) the label does not appear to influence the rate of water exchange and2) estimates ofP f of cell membranes measured with osmotic gradients are from 3 to 10 times greater than tracer permeability to labeled water (Pd) (38). The explanation for this observation can be understood in terms of the Fick relationship, which indicates that the rate of water diffusion should be directly proportionate to the concentration gradient between compartments. After instillation of HOH and3HOH into the air spaces, the concentration of water in the air spaces is 55 mol/l, whereas that in the vasculature is only 0.3 mol/l less (54.7 mol/l). There is consequently considerable back diffusion of HOH molecules immediately after instillation of the water. In contrast, there is initially no3HOH in the perfusate, and consequently equilibration of 3HOH proceeds much more rapidly than that of HOH.
As indicated in Fig. 1, C3 hoh fell to values close to those of Choh after 120 s. This suggests that samples collected at these times are derived from exchange vessels with slower flow, in which the removal of HOH as well as3HOH is limited by capillary flow rather than by diffusion.
(Pf S)Aand (Pf S)endo
Calculation of filtration coefficients of the pulmonary membranes are complicated by uncertainties concerning the surface area involved in these experiments. Although estimates are available concerning the total endothelial and epithelial surface areas of intact lungs (39) and of alveoli near the surface of the lungs (2), there is no way to be sure either what fraction of the alveoli are filled with fluid or whether they are uniformly perfused. We have therefore chosen to express our data in terms ofP f ×S products (in cm3/s, Fig. 3). For the control experiments (group 1), (P f S)A= 5.77 ± 0.85 cm3/s. If a value of 6,080 cm2 is assumed forS of these lungs (37), then would be equal to 9.4 ± 1.4 × 10−4 cm/s. This value is significantly less than that estimated from fluorescent studies in mouse lungs by Carter et al. (2) (0.17 ± 0.001 cm/s at 25°C) and may reflect differences in the surface area actually perfused, the species studies, and the experimental approaches used in these studies. It is possible that the capillaries near the surface of the lungs visualized by Carter et al. are more permeable to water than the remainder of the vessels of the lungs or that the vessels in the mouse lungs have become more permeable because of experimental conditions.
Values of (P f S)Awere increased by a factor of 2 when perfusion rates were increased by a factor of 3.4. This increase could reflect an increase in the effective surface area of the vascular bed because of the tendency of increased intravascular pressures to recruit additional vessels or distend vessels that are already perfused. (P f S)Awas also increased when either 77 or 308 mM NaCl was instilled into the air spaces rather than water. It is possible that instillation of water into the air spaces causes sufficient cellular swelling to reduce local perfusion and the surface area of exchange. Cooling the perfusate, instillate, and lungs to 11°C did not have a significant effect on either C3 hoh or Choh compared with experiments conducted at 37°C but did reduce the peak Choh-to-peak C3 hoh ratio by 25%. Calculated values of (P f S)Awere not significantly different from those of the control experiments. This relatively small effect of cooling is consistent with an activation energy of <4 kcal/mol and is compatible with movement through pores rather than lipid membranes (38).
Mercurials inhibit transport through both aquaporin-1 and -5 channels. The action of HgCl2 appeared to have a much greater effect on filtration through the endothelium than through the alveolar-capillary barrier as a whole (Fig. 3). The simplest explanation for this observation would be that1) there are more water channels on the endothelium that can be inhibited by HgCl2 than those that are present on the epithelium and 2)P f Sproduct of the epithelium is significantly less than (P f S)endo. (P f S)endovalues were greater thanP f Svalues of the epithelium in these experiments but, once again, direct comparisons are difficult to make because of the different experimental approaches used in these studies, including different methods of calculating filtration in the fluid-filled and air-filled lungs (see ).
Evidence for inhibition of filtration between the vasculature and air spaces of lungs by HgCl2 has been reported by Folkesson et al. (12) and Carter et al. (2). No attempt was made in these experiments to distinguish between endothelial and epithelial effects of this inhibitor. Folkesson et al. (12) found that HgCl2 slows water movement across the cell membranes of type II cells. Because these cells cover <5% of the alveolar surface, it cannot be concluded that aquaporin-1 in these cells represents an important route for water transport across the alveolar-capillary barrier.
Distribution of Aquaporin-1 in the Alveolar-Capillary Barrier
Confocal microscopy showed the presence of aquaporin-1 antigen along the endothelium, but it was not possible at the resolution of these studies to tell whether there was any present in the epithelial cells. The immunogold studies demonstrated aquaporin-1 along both the basolateral and apical surfaces of the endothelium. Lesser quantities were observed on some of the type II cells, but none was associated with the type I cells, which cover most of the epithelial surfaces of the alveoli.
Evidence for the presence of aquaporin-1 on continuous endothelial cell membranes has been reported by a number of laboratories by using either in situ hybridization or specific antibodies (17, 37, 40). Schnitzer and Oh (31) found that expression of this molecule on the luminal surfaces of rat lung endothelium is comparable to that present on rat blood cell plasma membranes. Concentrations of aquaporin-1 were particularly high in the caveolae of these cells.
Although there appear to be relatively few aquaporin-1 molecules in the epithelium, other aquaporins such as the mercury-insensitive water channels (16) and aquaporin-5 (28), which is sensitive to HgCl2, may promote the movement of water between the air spaces and the vasculature of the lungs. The relatively slight effect that cooling has on the transport of HOH relative to 3HOH from the air spaces to the perfusate that we observed in the group 4 experiments is consistent with the presence of such channels. It is therefore likely that much of the osmotic water transport between the air spaces and perfusate occurs through some sort of channels, though there appears to be less aquaporin-1 in the epithelium than the endothelium.
Effect of Osmolality on(PfS)A
When 77 mM NaCl rather than water was instilled into the lungs, the calculated value for (PfS)A exceeded that observed after instillation of solute-free water (see Fig. 3). This difference in the calculated value of (P f S) may better reflect tissue perfusion (increasedS) rather than a difference inP f because the average peak CTHO was greater after instillation of 77 mM NaCl than after instillation of water, although this difference was not quite significant (Table 2). (P f S)Awas also greater when hypertonic saline rather than water was instilled into the lungs, causing water to move from the perfusate to the air spaces. Once again, this may indicate better perfusion because the average peak C3 hoh may have been greater when hypertonic saline rather than water was instilled into the lungs. It is possible that tissue perfusion was reduced by cellular swelling after instillation of solute-free water (see below).
As indicated in Fig. 5, air-space [Na+] had reached 124 meq/l 4 min after instillation of 5 ml of 77 mM saline into the air spaces. This is equivalent to the loss of 1.59 ± 0.08 (SE) ml from the air-space compartment and is ∼0.3 ml greater than that which reached the perfusate, suggesting retention of a small volume of water in the tissues. When 5 ml of 308 mM NaCl were instilled into the air spaces, concentrations in the air spaces fell to 183 meq/l when air-space fluid was collected after 4 min. This is equivalent to the movement of 3.43 ± 0.26 ml of water into the air spaces, a value that did not differ significantly from that which was lost from the perfusate.
Movement of Solutes After Osmotic Challenges
After instillation of water into the air spaces, decreases in [Na+] and [K+] were initially similar to decreases in protein concentrations in the perfusate leaving the lungs (Fig. 4 A). As in our previous studies, this suggests that ςf values of the endothelium and epithelium to these solutes are ∼1.0 (7, 11). It is difficult to compare this ςf value with previous estimates of solute reflection coefficients (34, 26) because the latter values were on the basis of what may be inappropriate comparisons of changes in weight induced by increases in hydrostatic pressures, which tend to increase interstitial volumes, and decreases in weight caused by hypertonic infusions, which decrease cellular volumes (7).
After instillation of water into the air spaces, [K+] in the perfusate initially fell by approximately the same amount as [Na+], osmolality, and concentrations of EB-labeled albumin. However, [K+] subsequently increased relative to [Na+], concentrations of EB-labeled albumin, and osmolality. Because there were no red blood cells in this preparation, it can be concluded that aspiration of water resulted in the release of K+ from the pulmonary tissues. This could be related to normal volume control mechanisms that result in loss of K+ from cells after cellular swelling in hypotonic media (33) or to cell damage caused by this very hypotonic exposure. Movement of K+into the perfusate was not observed when less hypotonic solutions (77 mM NaCl) or hypertonic solutions (308 mM NaCl) were instilled into the lungs or when the lungs were cooled to 11°C. However, considerably more K+ entered the air spaces when 77 mM NaCl rather than 308 mM NaCl was instilled into the lungs (Fig. 5). Rapid increases in air-space [K+] have been observed when a variety of isotonic solutions have been instilled into the lungs (6, 8), and [K+] in the alveolar epithelial lining fluid is significantly greater than in plasma (23). It is therefore likely that factors in addition to cellular swelling are involved in the movement of K+into the air spaces. Nevertheless, considerably less K+ entered the air spaces when hypertonic saline was instilled into the air spaces, suggesting that cellular dehydration may have reduced K+ losses from pulmonary tissues.
Role of Aquaporin-1 in Osmotic and Hydrostatic Movement of Water in Lungs
Both the physiological and morphological data obtained in this study are consistent with the presence of more aquaporin-1 on the endothelium than on the epithelium. This distribution of water channels would tend to link the osmolality of the subepithelial tissues more closely to that of the blood than any solutions that may be aspirated. The presence of fewer aquaporin-1 molecules in the epithelium may also slow evaporative water losses from the alveoli. However, recent evidence suggests that the absence of aquaporin-1 from red blood cells does not result in any clinical disorders (27), and the exact role of this membrane protein remains to be determined. It is possible that, in some tissues, water conduction can occur through alternative aquaporins. Some water transport across the epithelium may also be mediated by aquaporin-5 channels, which are sensitive to HgCl2 and have been detected in the type I epithelial cells.
Aquaporin-1 is found in considerable abundance on many epithelial surfaces that conduct large amounts of water, such as the renal epithelia of both proximal tubules and the thin descending loop of Henle (37), the colonic crypts, choroid plexus, ciliary body, iris, sweat gland, and gall bladder (15, 16, 37, 38). However, as indicated above, aquaporin-1 also appears to be present on most continuous capillary beds (e.g., lung, skeletal muscle, and heart) aside from the brain and is present on both the basolateral and apical membranes of these cells (31). In contrast, it is absent on discontinuous endothelial membranes, such as those of the liver (31). Although the present study is consistent with the conclusion that some of the water crossing the endothelial barrier in response to osmotic gradients traverses mercury-sensitive aquaporins, it cannot be concluded that these structures limit the rate at which filtration occurs across the capillary walls in response to increased hydrostatic pressure gradients. Because water moving through aquaporins is not accompanied by solute, filtration through these channels would be associated with the development of an osmotic gradient that would slow further filtration. Thus the rate of filtration that occurs when intravascular pressures are increased may be limited by the rate of solute movement rather than the number of aquaporins present or their permeability to water.
The expert technical assistance of Joanne McCormack and Karin McCarthy is gratefully acknowledged.
Address for reprint requests: R. M. Effros, Medical College of Wisconsin, 9200 West Wisconsin Ave., Milwaukee, WI 53226.
This study was supported by National Heart, Lung, and Blood Institute Grants HL-18606 and HL-25822.
Results of this study have also appeared in abstract form:FASEB J. 9: A279, 1995.
- Copyright © 1997 the American Physiological Society
Calculation of the Pf S Product of the Alveolar-Capillary Barrier [(Pf S)A]
Unless information is available concerning the osmolality of perfusate in the pulmonary exchange vessels, no estimate can be made of theP f Sproduct of the membranes separating the alveolar and vascular compartments. Because the perfusate leaving alveoli that contain the solution instilled into the air spaces becomes mixed with perfusate from air spaces not exposed to this solution, it cannot be assumed that the outflow osmolality (cv) is the same as the osmolality of the fluid leaving the pulmonary capillaries (cvasc). Three factors could contribute to differences between the osmolality of the perfusate collected from the venous outflow and that of the perfusate leaving the capillaries: 1) the outflow from some regions of the lung that became filled with the instilled fluid may have not reached the venous samples at the time they were collected; 2) some alveoli may not been filled with fluid; or3) the water from some portions of the lung may have been absorbed into the vasculature at an earlier time.
To calculate the average osmolality present in the vessels that exchanged water with the air spaces, the model shown in Fig. 9 was used. As indicated in this figure, it was assumed that both3HOH and HOH entered the exchange vessels of the air spaces represented at thetop of the diagram. It was assumed that there was no exchange of 3HOH or HOH in the remaining air spaces (bottom of the diagram). As explained in discussion, it was assumed that3HOH concentrations in the perfusate leaving the exchanging alveoli were the same as that in the air spaces (cas) Equation A1 Once3HOH leaves the microvasculature, it becomes mixed with perfusate that has not been exposed to the instilled fluid. Consequently, the concentration of3HOH in samples of the venous blood collected from the left atrium (i.e., [3HOH]v) is less than that in the microvasculature or air spaces. If conservation of 3HOH is assumed Equation A2 Furthermore, if conservation of the solutes in the pulmonary vasculature is assumed Equation A3 CombiningEqs. EA2 and EA3 yields Equation A4where ca designates the osmolality of the perfusate entering the pulmonary arteries.
Once ca and cvasc are known, the average osmolality in the capillary was calculated by an approximation, by which it was assumed that flow was constant between the arteries and veins. Under these circumstances, the average osmolality of the perfusate in the capillaries (cc) falls exponentially (5) and can be calculated from the equation Equation A5 The assumption of an exponential decline in concentration represents an approximation in concentrations because there is a small change in the flow that occurs between the artery and veins, a factor that was not considered in the model of Crone (5). The value of (P f S) was calculated in Eq. 14 .
Calculation of the PfS Product of the Microvascular Barrier [PfS]endo
Calculation of theP f Sof the pulmonary exchange vessels was based on1) the rate of movement of water (Choh) from the capillaries to the lung tissues after 0.7 ml of water had been introduced into the arterial inflow and2) the concomitant difference in microvascular and tissue osmolalities, Δc. As described inmethods, both the perfusate and injection solution contained EB-labeled albumin. In contrast, only the injection solution contained [14C]dextran and3HOH. A simple model of the lungs was considered in which the extravascular space of the lung was considered to be a single compartment in which concentrations fell uniformly as some of the injected water entered the lung tissues. Because the transit times of the water and each of the indicators through the lungs were heterogeneous, the venous samples contained a mixture of perfusate that contained the injected-water bolus and perfusate that did not contain the injectate water. Venous concentrations, consequently, did not equal concentrations in the capillaries. However it was assumed that the fluid flow of water out of the vasculature (Choh) was reduced by the same fraction as the osmotic gradient (Δc) responsible for this flow. For the sake of simplicity, no attempt was made to correct for this dilution, and the rate at which water had been cleared from each sample was divided by the osmotic difference calculated from the venous osmolalities.
Because concentrations of EB-labeled albumin in the injection solution ([EB]inj) equaled those in the perfusate ([EB]p) arterial concentrations must have equaled those in the perfusate: [EB]a = [EB]p. In contrast, there was no [14C]dextran in the arterial inflow, and concentrations of the injected solution were diluted after injection into the pulmonary inflow. If it is assumed that there is no significant loss of either the EB-labeled albumin or [14C]dextran in transit through the pulmonary vasculature, then the concentration of [14C]dextran in the perfusate entering the arterial end of the microvasculature ([dex]a) can be calculated from the concentration of [14C]dextran found in the collected venous samples ([dex]v) by using the equation Equation A6After injection of a volume (Vinj) of 0.7 ml of water into the pulmonary artery, there is a decrease in the osmolality of the perfusate (cp) to ca. Following is the calcualtion for ca. It is assumed that there is conservation of osmoles and volume in the inflow tubing, and then Equation A7 where Va and Vp are volumes of water in perfusate and pulmonary artery, respectively.
Furthermore, it is assumed that there is conservation of [14C]dextran in the injection solution Equation A8 where [dex]inj is the concentration of dextran in the injectate.
Combining Eqs. EA7 and EA8 yields Equation A9An estimate of the average osmolality of perfusate in the capillaries (cc) was made by assuming that there was an exponential increase in osmolality between the arterial and venous ends of the capillaries Equation A10Calculation of the osmolality of the tissue (cti) was based on a simple model in which it was assumed that, on entering the capillaries, concentrations of 3HOH in the perfusate immediately equilibrated with that in the tissue and [3HOH]c= [3HOH]ti. The concentration of 3HOH entering the capillaries ([3HOH]a) can be calculated from the equation Equation A11 The decrease in 3HOH concentrations between the artery and capillary were determined by the relative volumes of the capillary space (Vc) and the extravascular tissue (Vti) Equation A12These equations assume flow-limited distribution of3HOH in the tissues, with instantaneous equilibration of3HOH with the extravascular compartment of the lungs. The change in the tissue osmolality relative to that of the capillary that was due to the flow of water from the capillaries to the tissue should be inversely proportionate to the size of these compartments Equation A13where cti, ndesignates the osmolality in the tissue perfused by thenth sample, and cti, n −1represents the osmolality of the tissue perfused by the previous sample. Then Equation A14 The flow of water from the capillary into the tissue was estimated from the equation Equation A15 and theP f Sproduct was then determined (see Eq.15 ).


text:
Friday, December 9, 2011
Torkil Gudnason / Dogs with Wigs
23 comments:
- Daniela said...
I feel bad for these dogs. It doesn't make this any less hilarious though.
- December 9, 2011 at 10:25 AM
- Lorena said...
I love these pictures !
AND if dogs and owner are alike.... well mine are quite crazy and loud.
- December 9, 2011 at 10:38 AM
- This comment has been removed by a blog administrator.
- December 9, 2011 at 10:41 AM
- Dancing Branflake said...
Oh dear. I am sitting here very uncomfortable. Ummm.... I swear one day we'll have to answer for the humiliation we put dogs through. ps.. love your tags. You are such a Gemini. Wait, are you?
- December 9, 2011 at 10:44 AM
- Heidi said...
These photos are great. I like how the dog on the right in the top photo is showing us just exactly how much he's enjoying wearing his wig.
- December 9, 2011 at 10:53 AM
- Trissta said...
LOVE. This reminds me of those crazy segments on Sesame Street back in the day with the dogs that were dressed up in human clothes and human hands. lol crazy crazy. I totally agree that owners are usually reflected in their pets and vice versa.
Much Love,
- December 9, 2011 at 10:57 AM
- Caroline said...
So flipping awesome!!!
- December 9, 2011 at 11:02 AM
- Kitty Stampede said...
You really cannot get any BETTER than dogs wearing wigs. you just can't.
these are freaking awesome photos!!!!
thanks for the giggles!!!!
I would love to do this.
- December 9, 2011 at 12:12 PM
- Annabelle said...
yah....Im gonna have to go with No on this one.
That boy pug's little red lipstick is peeping out at me and its creeping me out.
Wigs creep me out too, so it's just too much uncomfortable for my taste.
- December 9, 2011 at 12:58 PM
- Lara said...
It's like they were always meant to have hairstyles! Why don't dogs always look like this!!??
- December 9, 2011 at 1:10 PM
- Aline said...
these are so rad. i would hang the first little puggy one in my home in a heartbeat!
- December 9, 2011 at 2:03 PM
- ajg-jane said...
The poodles look like it's just another day wearin' a wig.
- December 9, 2011 at 8:56 PM
- Stem Cell Therapy in Dogs said...
Such a nice photos. It is really difficult to have a photo shoot on dogs there are lot of things to consider and those dogs were so funny.
- December 10, 2011 at 4:19 AM
- bananas. said...
ahahahahahahahaha the first two pugs are cracking me up! did you notice his peen in the first pic. ewwwww!!! if only dogs had hair like this...and hair stylists too! lol.
- December 10, 2011 at 1:47 PM
- ANDWHATELSEISTHERE said...
Okay, these dogs have better hair that I do. Talk about a social living standard here, hello?
- December 11, 2011 at 3:09 AM
- fabulousjunk said...
omg... reminds me of something my mom would do to my dog. haha. Oh by the way, that comment you left me about Van Halen had to be the best comment ever made on my blog. Seriously, that is so awesome! I'm glad I feel a little less alone about my groupie fantasies. :)
- December 11, 2011 at 9:59 AM
- Miss Caitlin S. said...
that's really funny, I like these a lot. And interesting that there's not much difference between the models and dogs.
- December 12, 2011 at 1:15 AM
- Kristine said...
Those are some good looking dogs! I am such an easy mark, take a photo of a dog doing anything and I am happy.
- December 12, 2011 at 3:51 AM
- SabinePsynopsis said...
Great photos. You know, I can TOTALLY see the owners when I look at the dogs :)
- December 12, 2011 at 8:36 AM
- Claire Kiefer said...
Hilarious. The poodle with the white fro is particularly funny!
- December 12, 2011 at 12:51 PM
- melifaif said...
LMFAO! The 2nd one reminds me of my daughter...lol! Only I can say this...but don't tell her....
- December 15, 2011 at 12:42 PM
- Down and Out Chic said...
OMG, hilarious! these dogs are good sports because my dogs would bit me if i tried to put a wig on them. ha!
- December 20, 2011 at 3:33 PM
I can't never imagine dogs with wigs, i love the last one it cute the rest they're ok. thanks for sharing anyway.
- March 27, 2012 at 2:30 PM

subset: commoncrawl


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|Ocean View, HI
Are you thinking of moving to the Big Island of Hawaii, even perhaps from another island? If this is the case then I know that you will have concerns about moving to the Big Island. Concerns that range from affordability of rental units, cost of living, schools, medical, social, security, weather and the list is long and different for each person considering a move. This blog post is going to deal with some of the basic concerns at the same time I will highlight some of the experiences my family and I have had in the 2 years living on the Island. Much of this article is true for all the Hawaiian islands but these are my impressions on the Big Island specifically.
|Wiapio Valley
|Pine Trees Beach, Kona
First of all you must know that Hawaii is made up of a chain of mostly dormant volcanoes and six livable islands located in the middle of the Pacific Ocean. It is probably the most remote place in the world. Flying here from any location, be it the USA West Coast, Canada, Alaska or the Far East, will not take less than 6 hours. However the islands are not remote by any means in that they are all serviced by a network of major airlines from all over the world and then smaller airlines or commuters will get you around the islands. It takes about an hour with Hawaiian airlines to travel from Hilo to Honolulu, the major city on the island of Oahu. Currently there are no sea-going ferries between islands which leaves flying as your only option. The cost of flying to the islands varies from airline to airline and time of year. I have seen mainland prices as low as $300 return and inter island prices can be as low as $50 so shop around on the internet.
|Macadamia Farms, with Volcano smoke in distance.
|Hawaiian Airlines
All of Hawaii's volcanoes are dormant, with the exception of the Big Island, where one of the 5 is currently active. Before moving to Hawaii I had never been on a volcano and just the thought of it conjured up images of life threatening lava consuming everything in its path, with mountains of exploding fire and brimstone raining down while pressing the fleeing, terrified population into the sea. While this is possible and true for many volcanoes, the active volcano Kilauea, is a rather innocuous affair and offers visitors an opportunity to enjoy and experience one of natures truly spectacular wonders. And indeed when the volcano shows signs of activity, our tourism rates increase. The lava currently flowing out of the Puʻu ʻŌʻō vent is heading downhill so slowly that it is cooling and solidifying, and as such is no concern for the small village of Pahoa in its path. I live about 8 miles from the Puʻu ʻŌʻō vent and last year I was able to sit on my porch at night and watch the glow of lava while sipping wine with good friends. Occasionally it is possible to smell the sulphur dioxide known as "vog". However when it is prevalent, it is more of a problem downwind with the vog being carried as far away as Honolulu. As it spreads it is normally no more annoying than a mild case of city smog. Hay fever sufferers do complain more at these times though.
|Halema'uma'u Lava Lake, 2015
There are numerous factors which should affect your decision about choosing the right location to live on the Big Island. Your age should be considered first. The island is by no means small and covers some 4000 square miles. It can take 4 hours or more at 55 miles per hour to circumnavigate the whole island. Therefore there are many cities and smaller villages to choose from. Some are remote and require more driving to access medical and other amenities. You will also find that as you search in the more remote areas, so the population becomes more ethnic Hawaiian, with people having lived in these areas for many generations. Depending on your character and as the new comer into a smaller village, you would need to decide whether you could fit into these villages. There are two main cities namely Kona and Hilo with populations accustomed to tourism, and therefore also to new comers on the islands. The 2 larger cities have populations of around 100,000 each and offer everything that any large city could possibly offer. Generally within 15 miles or so of Kona or Hilo, you can find reasonably affordable housing with good schools , colleges and access to medical, malls and super markets. One excellent point is the farmers network and access to farmers markets. There are many garage sales available over weekends to making life more affordable. If you are an older person, then medical access should be considered. The further away you reside from Kona or Hilo, the more difficult it would be for you in terms of emergencies and ambulance transfers. The islands do have Air Ambulances into Honolulu where medical facilities and medical care is world class.
|Observatory on Mauna Kea
Generating income would need to be your next deciding factor in choosing where to live. If you are an entrepreneur type, then making a living can be easy here. Many people make their livelihood by selling home products, running organic farms to selling at farmers markets. Others are service types such as plumbers and electricians; there seems to be no shortage of work for these professions. If you are a nurse or EMT, work opportunities are endless. In short, if you are motivated then you will find work. I work for an Air Ambulance company, my wife is a CNA, my son works with tourists on the Zip lines, and my daughter is a Life Guard and college photographer. Living closer to Kona or Hilo allows for more opportunity if you are a services industry provider. Tourism and farming can be further out so it all depends on what your needs are. In my line of work I am able to live about 30 minutes from Hilo. I get the comfort of country living while working in the city, the best of both worlds. Remember to do your due diligence in whole because a seemingly small thing like internet availability could be an expensive or slow service in a remote location. This would be especially important if your business relies on internet connectivity. For a short time I lived near Hilo and had good access to fast, cheap, unlimited internet. But when I moved out to the country, my internet bill tripled in price and it was capped. For me this was a small concern since my business is not internet dependent.
|Rainbow falls, with rainbow. Hilo
The Big Island has 13 climate zones. This means that you can enjoy rain forests, a desert, alpine and even snow on Mauna Kea and Mauna Loa volcanoes which rise to nearly 14000 ft above sea level. We are fortunate in Hawaii due to the trade winds, to have a moderate range in our diurnal temperature at sea level, which seldom rises beyond 95° summer and less than 65° winter but the humidity can be quite high. However as you climb up-slope the temperature and humidity drops and this can be a deciding factor when choosing where to live. A good example of this would be living in Hilo or Kona which are both sea level cities and can get to 95 degrees with high humidity opposed to living on the up-slopes of Kona at the 1500ft level or in Mountain View at 1500 ft level on the East side. The small increase in altitude can be more comfortable but accepting that you will be 20 minutes from town during peak traffic. East side rainfall is high at about 160 inches per year opposed to dry on the West side. The West side does get good rains at about the 1500 ft level. Generally there is no shortage of water but if you intend to run a small farm, you may want to consider water availability. Some farms and rural homes are on a water catchment process while others are able to connect to county water. I am comfortable with my catchment tank. The water runs through a filtration system to my house and is crystal clear.
|Green Sea Turtle, Kona
Another concern is the frequency of hurricanes which pass the Big Island. When we moved to Mountain View our house was in the path of Hurricane Iselle, a category 1 hurricane. Many people sustained damage to their property when trees fell down but fortunately we escaped unscathed. Much of the hurricane damage can be attributed to the invasive type of tree that is growing on the Big Island, and not really from the hurricane itself. To clarify, if the type of tree, an Ibizea from Africa, was not the dominant tree in the lower Puna district, the damage statistics would have been much lower. The county now has a program to remove these trees. These invasive trees grow very quickly in this high rainfall region but due to the lava also have shallow roots.
|Orchid Isle
The East side also has an invasive ant from Brazil. It is a tiny and almost unnoticeable ant called a Fire Ant. They normally form a line when they are on the move, making them easy to detect. These ants were confined to the lower shoreline of the East side but during hurricane Iselle, the wind must have blown many uphill where they have formed new colonies and spread. When they sting or bite, the victim experiences a slight burning itching sensation in that area of the skin. The sensation is not as irritating as those stings experienced with the bigger Fire Ant found on the mainland USA. I have managed to eradicate the ants on my property with an aggressive spray and bait program.
|Rainbow Isle
Dengue Fever has also recently been reported; it seems to be island wide rather than state-wide and it was probably imported by an infected visiting person. To date there have been 250 reported cases over the last 6 months when it was first detected. It is passed to humans from mosquito bites. Dengue Fever is treatable but more importantly, it is preventable by a regime of keeping the mosquito population breeding sites down. This is done by removing standing rain water sites like old tires from your locality and spraying plants with a solution of soapy water. Water catchments can be treated with chlorine. It is recommended to wear long sleeve shirts and trousers in the evenings and mornings when the mosquitoes feed and to keep your house screens on and repaired. Symptoms include headaches, rash and a fever of 104. The State has announced a spraying and education program and are determined to eradicate the disease. I recently heard on the radio that the spread and number of cases has declined substantially.
|2 Step, Place of Refuge, Captain Cook
Rat Lungworm causes eosinophilic meningitis. A very nasty little parasite that can adversely affect your life is found in Hawaii and particularly on the Big Island. If you are considering moving to Hawaii, then I would strongly advise you study up on this subject. In short the parasite is found in slugs and snails and perhaps even in their slime trails. Small pieces of slug can have many thousands of parasites. If you are coming to the islands whether as a visitor or moving here, this will affect you in some way. You will either be visiting restaurants, or buying fresh produce from the local market, or producing your own home grown produce. You must know then that your mere presence on the islands means that you are also a potential victim to Rat lungworm parasite. You must take this seriously. Small changes in your lifestyle will protect you. You can discourage the slugs from being on your property by destroying them when you see them, however wear gloves when you handle them. Check your garden frequently and especially your fruit and vegetables before you bring them into your house. Slugs can be so small that they are barely recognizable. You do not want them in your kitchen. Be especially cautious of cold vegetables or salads when they have been provided, unless you are absolutely certain of the source of the cold foods. This is true for restaurants.
|Hawaiian Sailing Canoe
Buying a home or choosing to rent. Unless you are completely knowledgable with the Big Island, I would highly recommend that you start your life here by renting a home. Give yourself time to get to know the island and communities before you settle on a house and location. When we moved here we rented a very nice 3 bedroom house above Hilo, and paid $950 per month rent for 8 months. The house was freshly painted with an attached garage and new carpets, so you do not have to settle for less. I recommend signing a 6 month lease if you intend to eventually buy. After 6 months your rent normally goes onto a month to month agreement anyway and you may just find a house and area you like in that time. I recommend that you use the service of a reputable real estate agency with a house inspector working for your interests before signing on a house. Like all places in the world, even new houses are prone to problems and a house inspection will pick up any defects prior to you signing. Its costs about $300 for an inspection and again check the real agency and inspectors for reviews before settling for his or services. Buying on the Big Island does allow you to get more bang for your buck and certainly as you get further away from either Hilo or Kona, you can find very attractive houses at very affordable rates with good sized yards or land. However if you are buying or renting, be sure to check the time it would take for you to commute to work. If renting then check the house for all the obvious problems which could cost you later on vacating the property if these were not included in the itemized defects list normally supplied by the renting agency. There are some traps which you should check before signing to buy or rent, which could definitely make your stay an unfortunate experience. Depending on your attitude towards dogs, you must know that the neighborhoods do have caged dogs and I have actually seen a house with as many as 12 caged up. Some of these dogs are kept like this indefinitely on the pretense that they are used for security or hunting. These dogs are frustrated in their conditions and tend to bark incessantly. So check the houses around you. Revisit the area at different times of the day to see how noisy it is. Dogs tend to become quieter during the heat of the day, when its raining or when its very cold. Try to visit the house early in the day or later when the dogs are getting hungry. You will soon hear them. Roosters are also a problem and indeed there are whole rooster farms here. It goes without saying that you would want to avoid living near a rooster farm unless you are a rooster farmer. Many people are "Off Grid" in the surrounding country site, meaning that they are really off the grid with no electricity. Be warned though that people off the grid generally have to run a generator from time to time to recharge solar batteries. Both my adjacent neighbors run generators and at times it can be rather inconvenient. If you will be going off the grid, then generators shouldn't bother you.
When looking for a vehicle you will find the same opportunities through dealers or private sellers as you would anywhere in the USA. We brought our Chevy Suburban in from the USA mainland and it cost us about $2000 for shipping from Texas. If you are shipping from the West coast, it will cost about $1300 for a vehicle up to 21 foot in length. There are size dimensions which the shipping agencies work on so check with them. If you are buying through a private seller then expect island cars to have a little rust but surprisingly not as much as some states with salted roads. I find cars to be a little more expensive in Hilo than Kona and certainly it is cheaper to buy in Honolulu but you will pay about $450 for shipping and the air fare to and from Honolulu to view the car. The latter is more difficult to do unless you know someone in Honolulu who is prepared to screen a car for you. Otherwise go to Honolulu and buy through a dealer as they know the shipping rules and will assist with the shipping.
|Hawaiian Wa'a (canoe) race
Pets are welcome in the islands but because the islands do not have the rabies virus, the authorities want to keep it rabies free and have imposed measures for the importation of pets. Strangely if you are coming from the mainland USA, you can expect moving your pets to be a lengthy and somewhat expensive procedure (approx. $1000 excluding quarantine) which may include a quarantine period of 3 months( at $17 per day) . It seems easier to import pets from Australia than to bring your pet in from the mainland. However if you follow the inoculation procedure to the letter, you can avoid unnecessary headaches later on arrival. I recommend that you check from all sources before you jump on to the first plane to Hawaii with your pet. The inoculation procedure should start about 4 months prior to traveling. Certain exotic pets and animals are illegal here and again you would need to check with the correct authority.
|Hapuna sunset
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Concluding Thoughts
In looking at liberation theology and theology of hope through Moltmann and Gutiérrez, one can surmise that both are political theologies that attempt to orient theology towards the downtrodden of society, albeit from different contexts. There is also an undeniable common thread of Marxism to be found in both their political theologies. Moltmann himself notes that Marxism is used by most liberation theologians, including Gutiérrez, as "an analytical instrument – a way of grasping the situations of the poor in Latin America. . . . Put in simpler terms: Marxist analysis, yes; communist therapy, no!" (Experiences in Theology, 245). This use of Marxism, however, isn't necessarily true for every liberation theologian, as not all liberation theologians adopt Marxist theory, (e.g., Jon Sobrino seems to deliberately avoid the use of Marxian categories such as class struggle and a classless society in his writings; another example is Hugo Assman).
While both Moltmann and Gutiérrez have used Marxist social theory, both decidedly ditching Marxism's dialectical materialism and atheism, there seems to be a stark difference in how they base their use of Marxism. Gutiérrez appears to start with Marxist analysis and then seeks biblical verification, though naturally departing from Marxism when it comes to the obvious issues of death and God, etc. Conversely, Moltmann starts with the biblical depiction of the future kingdom of God and then employs Marxism as a critique on how the current way of things is opposed to this future hope. The primary appeal to Moltmann in Marxism (as he found it in Bloch) is its imagination for a better future society, rather than the Marxist strategy for proletarian revolution which seems to be a more useful aspect to Gutiérrez.
Gutiérrez's understanding and use of "hope" stands in distinction to Moltmann's; he has not just uncritically embraced Moltmann's view of hope but has instead only borrowed from it in order to create his own. Gutiérrez's hope is directed towards a liberating utopia and a plan for a qualitatively changed society that is more just towards the poor. This is different to Moltmann who sees hope as being founded upon the resurrection of Christ and directed towards the ultimum novum, declining to pivot around a more static final objective such as Gutiérrez's utopia. In short, one could perhaps say that Gutiérrez's theology reflects a temporal human-based hope, while Moltmann's theology employs a more future divine-based hope.
Moltmann's influence on Gutiérrez (and other liberation theologians) is a noteworthy case of theological cross-fertilization between first-world and third-world theologians. Yet, Gutiérrez was wary of not taking too much from Moltmann and Europe, primarily seeing the benefits of hope theology in its critiques of the hyper-individualized gospel and our overly capitalist society, while also coveting the stimulating effect Moltmann's theology has on political consciousness and engagement.
An adequate theological stance on hope must rest upon sufficient grounds. The eschatological hope of Moltmann that is predicated on the resurrection of Christ and the everlasting faithfulness of God appears somewhat different to Gutiérrez's more evolutionary optimism, that seems to depend on an extrapolation of the present process, thus providing a less sure hope. In the end, after briefly looking at the role that Marxism and hope play in their respective theologies, one could almost say, to borrow a wordplay from Kayayan, that Moltmann's theology of hope was effectively secularized in Gutiérrez's liberation theology, turning it from a theology of anastasis (resurrection) into a theology of epanastasis (revolution)! Naturally, though, Gutiérrez's revolution would be with bread and wine, not guns and bloodshed.


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How Rwanda is positioning its youth for the future
by March 16, 2016 12:49 pm0
Lydie Hakizimana, Private Sector Foundation's Youth Chamber President told CNBC Africa that the east African country, Rwanda, needed to capitalise on this demographic dividend.
"The biggest asset the country has is its youths; about 70 per cent of the country's young people are between the ages of 14 and 35 years," said Hakizimana.
"They are making a contribution in the economic growth story of the country through investments in agriculture, ICT and public service."
Hakizimana lamented the skills deficit calling on the government to escalate current efforts.
"Most of the youths today lack skills which is a huge challenge, but we should recognise initiatives being spearheaded by the government of helping to skill the unskilled," she said.
Rwanda, through the Economic Development and Poverty Reduction Strategy (EDPRS) project, seeks to skill youths in critical and scarce skills gap in the following high priority sectors such as infrastructure, agriculture, natural resources, health and education.
Hakizimana said access to funding has also been one of the problems that has been bothering young entrepreneurs.
"The challenge that youths are now facing is coming up with innovative ideas that are bankable. We currently have multiple sources of funding," she added.
There has been stigma in the past regarding certain sectors but this is changing largely due to the intervention of technology.
"More and more youths are looking in sectors previously unattractive, they are looking for ways to bring solutions through interventions of the ICTs," added Hakizimana.


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Click here to RSVP to the event on Facebook.
We Hunt Buffalo, Yawning Man, Black Pussy, La Chinga
Official Site: http://www.truckfighters.com/
Facebook: https://www.facebook.com/truckfighters
Youtube: https://www.youtube.com/user/TruckfightersTV
If you are interested in groovy, stoner-influenced, progressive and melodic hard rock you probably know about Truckfighters already. "The best band that ever existed" as Josh Homme said in the 2011 "fuzzomentary" Truckfighters. You might also know the story about how Niklas 'Dango' Källgren teamed up with bassist and vocalist Oskar 'Ozo' Cedermalm in a worn down rehearsal space in Örebro, Sweden in 2001 and how the first song they ever recorded, Desert Cruiser, since then has gone on to become a desert rock anthem. You probably also know that the band itself has gone on to release four full length albums and played sold out shows on four continents.
The band's upcoming fifth album, V, is released September 30th under exclusive license to the established label Century Media Records. "Century Media have been very cool with us. We've been able to keep that do it yourself-philosophy of ours, to focus solely on our creative process and producing the album", says Ozo.
We Hunt Buffalo
Official Site: http://www.wehuntbuffalo.com/
Facebook: https://www.facebook.com/wehuntbuffalo/
We Hunt Buffalo create songs with hard hitting, fuzz rock grooves and psychedelic overtones. It's a culmination of many years playing music together in a small Vancouver suburb.
Yawning Man
Official Website: www.yawningman.com
Facebook: www.facebook.com/yawningmanofficial
Influenced by everything from the psychedelic to the folkloric, jazz, hard rock, roots punk, world music, the sound of this trio while hard to describe is visual and cinematic , heavy and melancholy. driving & daydreaming.
Black Pussy
Facebook: https://www.facebook.com/lessinfomoremojo
Bandcamp: https://blackpussy.bandcamp.com/
Black Pussy is a group of rock n' roll pot-smoking fun-loving long hairs that are passionate about art, creativity and the dynamic human spirit which refuses to submit. We encourage everybody* to take the ride with us.
La Chinga
Facebook: https://www.facebook.com/La-Chinga-389979744359207/
Youtube: https://www.youtube.com/user/LaChingaVideo/videos
A hard rock power trio with psychedelic powers sitting on the worlds edge in Vancouver. Drawing from Sabbath, Zep, the MC5 and their own superbad habits, La Chinga has established a beachhead with Detroit's Small Stone records and a penetrating buzz across Canadian campus radio. Released in 2013, the bands debut self-titled album prompted ecstatic reviews from the kind of people who wonder why they don't make em like this anymore.
+19 Doors: 7:00 PM
$20 +S/C adv
Tickets also available at
Charge by phone: 1-888-732-1682


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Posted 12 Nov 2013
Link to this post
How can I use INPC function of devexpress in Justcode?
And please let me know the merits of Justcode than Coderush?
Posted 15 Nov 2013
Link to this post
You can make a code template as show here using the code below if you have already implemented the INotifyPropertyChanged interface.
Notice that the type of the property should be automatically evaluated, but right now we have some issue with the command.
public $3=GET_ELEMENT_TYPE($1$)$ $2=SUGGEST_NAME$
get {return $1=SELECT_VARIABLE()$;}
if($1$ == value)
$1$ = value;
if (PropertyChanged != null)
PropertyChanged(this, new PropertyChangedEventArgs("$2$"));
For more information about the templates visit our online documentation.
I am not aware of CodeRush so the the best way to evaluate will be to try both products and decide which one fits your needs best. Here is a link to JustCode page where you can download a trial.
Furthermore, this article will give you a brief view of JustCode key features
Posted 14 May 2014
in reply to
Link to this post


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Zimanek: Brother Jonas holds his own with anyone
Georgia forward Jonas Hayes has thrived for the Bulldogs this season despite coming to Athens in the shadow of his twin brother Jarvis. Jonas has averaged seven points and nearly five rebounds while playing 20 minutes per contest. He started seven games when the Bulldogs were riddled with suspensions earlier this season.
Dot Paul/Staff
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Learn More about Microsites
Includes devices, machines other than land or buildings that is used in the operation of construction. Activity in which a structure is constructed from materials to give beauty to the home or building you expect it to be.
A representation of the inside of a building or room, Different room requires variety of interiors that best suits it. Focuses on how a structure as viewed by the users, meets their needs and requirements.
A person qualified in the art of construction, He does a contractual work as well as his equipped work. He acts both as an architect and a civil engineer providing customer expected building services through its personnel.
Its Free! Promote your Product :)


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Contact Info / Websites
- Skype
- zakchaos
2015 Submissions
'Splorin Rated Stars Explore the depths, and collect random powerups Platformer - Other afishouttawater Rated Stars a fish begins a journey far away from home Adventure - Other 10 seconds till calm Rated Stars tap or click to defened yourself from advancing waves Shooter - Multidirectional Just One Banana Rated Stars Just One Banana. Platformer - Other Stupid Cupid Rated Stars That Stupid Cupid Left a buncha hearts and saws in the sky! Platformer - Other


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First Team News
Pellegrini: Hart has our trust
Just a day after receiving backing from England manager Roy Hodgson, Pellegrini too got behind the 26-year-old stopper.
"Of course he feels that he didn't do very well [against Bayern Munich], but I think he doesn't have any problem. He is continuing with the team and with our trust we hope he will recover from the game," said the Chilean.
"After the game I always speak with all the players. I spoke with Joe and we analysed what we did in the game.
"Joe knows he must improve the way he is playing, but he knows he has our trust."
City come up against Everton in the Premier League tomorrow, the only team who remain unbeaten so far this campaign.
Pellegrini admitted he has not yet decided on his starting eleven and is giving it a lot of consideration.
"I will decide tomorrow who will start, I don't tell you now that Joe hart will play tomorrow, because the players don't know yet," he said.
"I think he [Hart] is being pushed hard enough. Costel Pantilimon is a good keeper also; he has all the right attributes to play. He's working hard and I'm sure that if he must play tomorrow or another day, he will do well."
The manager also revealed that Jack Rodwell is back training with the first team and that apart from the long-term injured Martin Demichelis and Stevan Jovetic, he has a full squad to choose from for tomorrow's game.
"Jack just started working with the whole squad yesterday. He is fit, but he will not be included tomorrow, but he is fit," he said.
"Everybody else is fit, apart from Jovetic who has a couple of small problems."
The full press conference will be on CityTV this afternoon.


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1/1 The Causeway Strathfield South 2136 NSW
Affordable Living
Located in a quiet, sought-after street, within close proximity to Ford Park and the Cooks River Cycleway, You will find this low maintenance half house. Features include:
⢠Three spacious bedrooms
* Entertainers lounge with air conditioning
⢠Laminated floor boards throughout & tiles bathroom
⢠Eat-in kitchen
⢠Laundry & neat and tidy bathroom
⢠Located close to public transport, schools & Belfield village shops
Email this agent
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Strathfield Partners Real Estate
Strathfield NSW, 2135
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Real Estate For Rent in Strathfield South, NSW 2136 priced between $260 and $480


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These items are not for sale and the descriptions, images and prices are for reference purposes only.
You can reduce the number of items displayed by entering a keyword that must be included in the description of the item.
An antique turquoise set bangle, 15ct yellow gold with soft rose overtones, hallmarked to inner Craddis, the Victorian oval bangle claw set across top with round graduating sized turquoise of intense blue colour, slight variation throughout, claw set with
A 9ct rose gold bangle, twisted pattern with central cabochon turquoise. Diameter 6 cm. Weight 4.4g.
A Navajo silver and turquoise bangle. Diameter 6.2 cm. Weight 65.9 cm.
A large Asian bangle, silver set with amber, turquoise. Total weight 127g.
An enamel bangle by Hermes, styled with gold Hermes coach logo on a turquoise background, 1 cm width, 6.5 cm diameter
Native American Sedona Indian jewellery suite, by Zuni artist Annie Quam Gasper, composed of a silver cuff bangle and graduated disc necklace inlaid with turquoise, both signed.
An enamel and silver bangle by Hermes, styled with a geometric pattern in red, turquoise and blue, stamped + Q, made in France, 65mm diameter, boxed
An antique Etruscan revival bangle, turquoise and seed pearls in raised frames,hinged bangle with twisted wirework, gold bloom embellishments and bead granulations; 14ct gold. Weight 23.7g.
An Australian gold and turquoise bangle by Wendt, the circular bangle having engraved decoration and set throughout with turquoise, in rose gold.
A tribal Indian silver stone set bangle, hinged bangle set with turquoise and ornamental stones. Wt. 99.7g.
A Mexican silver and turquoise cuff bangle
A Mid-Victorian bangle, of articulated and hinged design, the front section of the hollow link bracelet with repousse decoration and set with pearls, turquoise and garnets. 18ct yellow gold. Weight 25.5 grams.
A Victorian turquoise, garnet and gold bangle, modelled as a coiled snake, having turquoise cut en cabochon set to the head and tail with garnet set eyes, mounted in 15ct gold.
A gem set horn bangle, silver set with pearls and turquoise.
Turquoise solitaire set bangle
A turquoise and silver bangle with safety chain.
Gold bangle set with turquoise
14ct white gold, turquoise and diamond bracelet, the oval hinged bangle is claw-set to the front with a line of seven graduated oval turquoise alternating between brilliant-cut diamonds flanked by diamond shoulders, the twenty-two diamonds together weighin
18ct gold and turquoise bangle, Tiffany & Co., of hinged design, the circular bangle of bamboo motif is embellished at intervals with buff top turquoise, inner diameter approximately 55mm, signed Tiffany & Co.
Silver and turquoise bangle
A Victorian gold and turquoise snake bangle, yellow gold, diameter 58mm.
Collection of various costume jewellery, including coral and jade brooch, Scottish brooch, turquoise brooch and other brooches, bangles and buckles in a ceramic box (some items as inspected). Provenance: Seale Family, Sydney
An Afghani stone set cuff bangle; inlaid with turquoise and coral, width 30 50mm.
Silver and turquoise bangle,
Two Mexican silver bangles. One set with malachite, the other with turquoise.
Victorian 15ct yellow gold turquoise bangle of the Etruscan revival period, c.1850's featuring 53 cabochon turquoise set into a dome and with fine applied rope work and decorative designs either side total weight 24.5 grams
A turquoise and silver cuff bangle.
An Imitation turquoise cuff bangle.
Vintage silver and turquoise set cuff bangle together with a silver filigree bangle
American Indian 'Zuni' silver bangle set with turquoise
A bamboo bangle; top applied with silver lattice and set with a cabochon turquoise to carved shoulders. Diameter 72mm. Wt. 22.89g
Gold bangle, 18ct yellow, gold hinge design with a central floral setting containing one pearl and six turquoise, clip clasp and safety chain, stamped '18K'. Weight 33 grams
Antique 9ct yellow gold bangle with disk, set with three 1.5mm turquoise & three coins 7.4gms
Woven silver (925) bangle with mother of pearl & turquoise total weight approx 56 grams
9ct yellow gold Victorian, turquoise and seed pearl bangle of a plaited design, hinged with safety chain weight: 11.0gms
An Art Nouveau tortoiseshell and gem set bangle, the wide tortoiseshell cuff with applied gold panels set with blister pearls and turquoise cut en cabochon, in 14ct gold.
Early pink gold and turquoise set bangle. Missing 1 turquoise. W27.4g
Turquoise and seed pearl bangle, the round hinged bangle of pierced floral design decorated throughout with turquoise cabochons accented by seed pearls, mounted in 14ct gold, inner diameter approximately 55mm, 47.5 grams, turquoise and seed pearl deficient
Victorian gold tone and turquoise hinged bangle the clasp in the form of a bunch of grapes and vine leaves
Garnet and turquoise bangle, set with five oval garnets highlighted by turquoise and cannetille decoration all around, mounted in 18ct gold, inner diameter approximately 6 cm, 50.7 grams.
A silver turquoise cuff bangle; 55mm tapered cuff bangle set with three matrix turquoise panels.
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GraphicRiver Research R Letter Logo 9691418
Category: Logo Template | Comment: ( 0 )
GraphicRiver Research R Letter Logo 9691418
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Think outdoor garden, and beautiful plants, ornamental trees, flowering shrubs and rolling lawns come to mind. The mind's eye may not immediately envisage concrete statuary or even benches, pavers, stepping stones and the like.
However, any garden worth its name – be it in a small backyard or a sprawling turf – is quite incomplete without attractive concrete artworks. While statues, animal sculptures, gnomes, fairies, gargoyles and birdbaths add to the landscape and look quite becoming, other concrete items such as benches, urns, plaques and pavers are quite elementary to the garden scheme.
Indeed, the lush foliage of a garden gets an added charm and texture with garden art and statuary. Most of these items are cast in concrete as it is sturdy and durable.
This brings us to the question of first making molds for these concrete casts. Which mold making materials will be suitable for casting concrete? Let's take a look:
Liquid latex rubber – This has emerged as the mold making material of choice for casting concrete garden articles. It is flexible, long-lasting, maintains details well and most importantly, can easily resist the


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Wal-Mart Has Made 'No Decision' on VHS17 Jun, 2005 By: Thomas A., Kurt I.
Wal-Mart said "no decision has been made" on whether the giant retailer will stop carrying videocassettes after the 2005 holidays, a claim made by studio sources (HMR, June 12-18).
While cassette inventories will continue to shrink as demand for DVD grows, there is no firm February 2006 cutoff, according to Wal-Mart spokesperson Karen Burk.
"We've not made a decision," she said. "We will continue to carry VHS as long as there is demand from our customers."
Burk did not say what percentage of the chain's home video inventory is VHS or how many cassettes will be left in stores after the holidays.
"We don't discuss inventory," she said.
Burk's comments came on the heels of news reports in Home Media Retailing and The Hollywood Reporter citing studio sources and store department managers who indicated the chain is planning to cut back on VHS in some stores in its September merchandise "reset," based on customer demand. The stories also reported that by Wal-Mart's February 2006 "reset," the store could eliminate VHS from its shelves altogether.
Wal-Mart did not respond to requests for comment at the time of the initial story.