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Platelet-derived growth factor-A (PDGF-A) and its cognate receptor, platelet-derived growth factor receptor-alpha (PDGF-Rα), are required for pulmonary alveolar secondary septal formation. PDGF-A null mice have normal appearing terminal sacs and a full complement of PDGF-Rα expressing alveolar cells at birth, but thereafter, the sacs do not undergo septation. Failed septation is accompanied by a nearly complete absence of alveolar mesenchymal cells, which normally contain smooth muscle alpha-actin (αSMA) and produce elastin (Bostrom et al., 1996, 2002). Others have proposed that PDGF-Rα-expressing alveolar cells are precursors of alveolar mesenchymal cells which contain αSMA and produce elastin (myofibroblasts, MF) (Warburton et al., 2006). Whereas it is clear that alveolar MF are important, it remains unclear how PDGF-A influences the proliferation, migration, differentiation, and gene expression of the MF.
Studies using PDGF-A null mice suggested that PDGF-A may exert one or more of four potential effects on alveolar MF (Lindahl et al., 1997; Bostrom et al., 2002). First PDGF-A may increase αSMA expression in MF. Second, PDGF-A may promote the migration of PDGF-Rα-expressing MF from the septal base to the tip, where they produce elastin. Third, PDGF-A could promote the proliferation of MF. And fourth, PDGF-A may stimulate PDGF-Rα-expressing cells to assume an “activated” phenotype, diverting them from a lipid-storage to an αSMA-expressing phenotype (as is observed in hepatic stellate cells) (Davis et al., 1993). These effects are not mutually exclusive because in the hair follicles, PDGF-Rα-expressing mesenchymal cells in the dermal papilla do not incorporate BrdU, whereas PDGF-Rα-expressing mesenchymal cells in the papillary sheath both incorporate BrdU and express αSMA (Karlsson et al., 1999).
The seminal studies by Bostrom et al. and Lindahl et al. suggested that PDGF-Rα expressing alveolar cells require PDGF-A to ultimately occupy their characteristic position at the tips of the secondary septa (Bostrom et al., 1996; Lindahl et al., 1997). However several details need to be further elucidated in order to understand how this requirement is fulfilled. For example, is the PDGF-Rα gene transcribed concurrently during αSMA and elastin expression by MF, or is PDGF-Rα primarily required for establishing sufficient MF in the proper location? Once MF are established, PDGF-Rα may no longer influence the synthesis of αSMA and elastin, which are hallmarks of the alveolar MF. Second, do PDGF-Rα-expressing MF populate the septal tips because of a proliferative advantage, do they migrate there from the base of the septa, or are both mechanisms involved? Third, does PDGF-Rα influence the propensity for the MF to assume a contractile, αSMA-producing instead of a lipid storage phenotype?
To address these questions, we hypothesized that if PDGF-A mediated signaling specifies a myofibroblastic phenotype, then spatial and temporal colocalization of its cognate receptor, PDGF-Rα, should coincide with specific myofibroblast products. We have studied the chronology and location of PDGF-Rα expression during secondary septal formation in mice with hemizygous expression of the enhanced green fluorescence protein (GFP) that is regulated by the endogenous PDGF-Rα promoter. We have also examined the proliferative index of PDGF-Rα-expressing and nonexpressing cells in the alveolar wall. Cells containing one functional allele of the PDGF-Rα gene are functionally equivalent to cells with two wild-type alleles (Hamilton et al., 2003). By examining the location of GFP-containing cells with respect to αSMA, which is most abundant in the terminal alveolar duct, we have been able to correlate a characteristic of the MF with potential PDGF-A-responsiveness.
Studies from our laboratory and those of other investigators suggested that when lung fibroblasts adopt a lipid-storage phenotype, they exhibit fewer myofibroblast-like properties (diminished tropoelastin and smooth muscle actin) (Macfarlane and Heaf, 1990; McGowan et al., 1997; Burgess et al., 2005). Thiazolinediones (TGZ) and other peroxisome proliferator-activated receptor-gamma (PPAGγ) agonists diminished proliferation, stimulate lipid accumulation, and reduce collagen gene expression (a myofibroblastic characteristic) in lung fibroblasts (Burgess et al., 2005). PDGF-induced cellular proliferation is inhibited by TGZ suggesting that the effects of PDGF are dampened after stimulation of PPARγ. Therefore we also examined whether lipid storage differed among interstitial pulmonary cells that had high versus low levels of PDGF-Rα gene expression.
MATERIALS AND METHODS
Characteristics and Breeding of Mice Bearing the PDGF-Rα-GFP Transgene
The mice were originally obtained from Dr. Philippe Soriano (Fred Hutchinson Cancer Ctr., Seattle, WA) and their characteristics have been described (Klinghoffer et al., 2002; Hamilton et al., 2003). In summary, homologous recombination was used to insert the enhanced GFP gene into the PDGF-Rα coding region. The insertion resulted in translation of the GFP protein rather than PDGF-Rα, and the expression of GFP is temporally and spatially identical to that of the endogenous PDGF-Rα gene (Klinghoffer et al., 2002). We used mice that were heterozygous for the GFP insertion so one allele contained the wild-type PDGF-Rα coding region. These mice are phenotypically identical to mice bearing two copies of the PDGF-Rα allele including their postnatal generation of alveoli. The mice were housed in Thoren cages in a temperature-controlled environment with a 12-hr light–dark cycle and provided food and water ad libitum. The protocol was approved by the animal use committees at the Iowa City Veterans Affairs Medical Center and the University of Iowa.
Preparation of Lung Tissue for Confocal Microscopy
At postnatal days 4, 8, and 12 mice were euthanized with ketamine and zylazine and their lungs were inflated in situ to total lung capacity with 1% low melting point agarose in 0.5% paraformaldehyde, 15% picric acid, 0.1 M Na Phosphate pH 7.0, while the carcass was maintained at 4°C. After the agarose hardened, the lungs were removed and their volume was determined by displacement (Scherle, 1970). The lungs were sliced into 1-mm transverse sections with a McIlwain tissue chopper, and fixed for 6 hr at 4°C in 0.5% paraformaldehyde, 15% picric acid, 0.1 M Na Phosphate pH 7.0. Following fixation, the agarose was removed by incubating for 1 hr in PBS at 50°C. The tissues were stored at 4°C in 0.05 M NaPhosphate pH 7.2, 20% sucrose, 10% glycerol, 0.1% sodium-azide. To prepare cryosections for immunostaining, the external liquid was removed, the tissue was embedded in optimal cutting temperature (OCT) medium, quickly frozen and 100 μm sections were cut with a cryotome.
Immunohistochemistry and Confocal Microscopy
After removing the OCT, the tissue was permeabilized in 0.3% Triton X-100, washed with PBS containing 0.2% bovine serum albumin (BSA), incubated overnight at 4°C with Cy3-labeled mouse anti-αSMA (clone 1A4), and washed. The tissues were counterstained with 1,1'- dioctadecyl-3,3,3′,3′-tetramethylindodicarbo-cyanine perchlorate (DiD, Invitrogen Molecular Probes, Carlsbad, CA) which stains phospholipid-containing membranes and mounted. Sections were imaged using a Zeiss LSM 510 scanning laser confocal microscope (LSCM) with excitation at 488, 543, and 633 nm, and emission band pass filters 505–530 nm, or 560–615 nm, and a 650 nm long pass filter. Z-stacks were obtained at a 1024 × 1024 pixel density, using a 3 μm slice interval and a pin hole of 1 airy unit, which allowed resolution of individual nuclei.
To identify neutral lipid droplets, fixed and permeabilized tissue slices were incubated with (E,E)-3,5-bis-(4-phenyl-1,3-butadienyl)-4,4-difluoro-4-bora-3a,4a-diaza-s-indacene (BODIPY 665/676, Invitrogen Molecular Probes) (Bedner et al., 2000). Cells of mesenchymal origin were identified using mouse anti-vimentin, followed by goat-anti-mouse Alexafluor 543, which allowed us to localize lipid droplets that were in mesenchymal cells. The BODPY and vimentin-stained tissues were imaged using the Zeiss LSM 510 and the same parameters were used to image GFP and smooth muscle actin.
Image stacks were analyzed using StereoInvestigator (MBF bioscience, Williston VT). The alveolar entry rings (AER) were identified by circular or oval collections of αSMA that led into an airspace (alveolus), which ended in a sheet of cells (identified by DiD staining) within the z-stack. Individual GFP+ nuclei within a core of tissue that included the entry ring (ending of the alveolar duct) and alveolar walls of a particular alveolus were enumerated using the optical fractionator probe and their distance from the AER was ascertained (Howard and Reed, 2005). Tissues were analyzed from mice at postnatal days 4, 8, and 12 and the number of GFP+ cells was expressed relative to the volume of the lungs, per mm3 of tissue, and was corrected for the VD of the alveolar walls (obtained by point counting). We also studied temporal changes in the colocalization of GFP+ nuclei and αSMA at AERs. To reduce the dependence of the measurements on the intensity of αSMA immunostaining, sets of P-4, P-8, and P-12 tissues were stained and imaged in parallel. Color information, which was collected in a RGB format was converted to HSI format to dissociate the yellow hue from the intensity of the red αSMA immunostain. The area of each AER was ascertained by manually tracing around the αSMA, which needed to occupy over 80% of the ring. The pixel area occupied by yellow (hue) within the entry ring (coalescence of green GFP and red αSMA-containing pixels) was calculated using IPLab (BD-Bioscience, Rockville, MD) and expressed relative to the pixels occupied by GFP+ nuclei, or to the entry ring-area.
Isolation of Lung Fibroblasts
Immediately after euthanasia on postnatal day 12, lungs from PDGF-Rα-GFP mice were perfused with saline, removed, minced, and digested in a Hank's balanced salt solution with 5 mM CaCl2, containing 250 U trypsin, 75 μg DNAse I, and 300 μg collagenase per milliliter. The minced tissue was incubated at 37°C, and at 10 min intervals the suspension of released cells was strained through 100 μm mesh, and made 10% in FBS. After complete dispersal, the cells were sedimented at 500g, resuspended, and erythrocytes were removed by density sedimentation through 40% Percoll. To select for fibroblasts, cells were plated in 100-mm diameter tissue culture dishes and allowed to adhere for 1 hr at 37°C. After washing to remove nonadherent cells, the adherent fibroblasts were released by brief trypsinization and gentle scraping, and collected by centrifugation. The fibroblast-enriched population was enumerated and resuspended in media containing 10% FBS and divided into aliquots of 5 or 10 × 105 cells.
Flow Activated Cell Sorting to Identify αSMA
Two slightly different procedures were used to prepare freshly isolated LF for flow cytometry. Group A (N = 2): 1 × 106 cells were fixed in 1% paraformaldehyde in 0.1 M sodium phosphate pH 7.2 for 10 min at room temperature. After extensive washing in PBS, the cells were permeabilized in PBS containing 0.1% saponin and then incubated with equal concentrations (1:100 dilution) of either monoclonal phycoerythrin (PE)-conjugated anti-human α-smooth muscle actin (R&D Systems, Minneapolis, MN) or the PE-conjugated-IgG2A (isotype control) for 45 min at room temperature. The samples in Group B (N = 3) were processed in the same fashion with the following exceptions: 5 × 105 LF were fixed in 2% paraformaldehyde for 20 min at 4°C. After permeabilization, the LF were incubated in Mouse-on-Mouse blocking reagent (Vector Laboratories, Burlingame, CA) for 1 hr at 4°C, followed by either anti-αSMA or the isotype control IgG for 45 min at 4°C. After washing to remove unbound antibody, the LF were resuspended in PBS. For both groups, flow activated cell sorting (FACS) analysis was performed at the University of Iowa Flow Cytometry Facility using Becton Dickinson LSR II or FACS DIVA instruments. After assessing forward and side scatter, using an unstained sample, gating parameters were set. Compensation for any overlap of the PE and GFP signals was carried out using either samples stained with the anti,-αSMA or Becton Dickinson comp beads (BD Biosciences, San Jose, CA), consisting of anti-mouse Igκ exposed polystyrene microparticles which bind mouse antibody, and a negative control of FBS-exposed particles. The beads were incubated with the anti-αSMA antibody, and thus were used to optimize for a positive and negative PE signal. An isotype control sample was used to correct for nonspecific binding of nonimmune IgG. Flow cytometry was performed using a LSR II or FACS DIVA (Becton Dickinson, Franklin Lakes, NJ) instrument. The data were analyzed using both CellQuest Pro software (BD Biosciences) and SigmaStat (Jandal Scientific, San Rafael, CA). Each “N” represents a separate experiment (litter of mice) and greater than 7000 gated events were analyzed for each “N.” After pooling the results from all five experiments, a two-way analysis of variance revealed no significant experimental differences between the Groups A and B (P = 0.265, Student-Neuman Keuls post-hoc test). Therefore, data from groups A and B were combined.
Evaluating the Purity of the Freshly Isolated Lung Fibroblasts
The putative fibroblast population was examined for contamination by endothelial and epithelial cells and macrophages using FACS. Cytotypic markers (CD31 for endothelial and F4/80 pan macrophage marker) were detected in an unfixed sample using a PE-conjugated monoclonal rat anti-mouse CD31 (BD Biosciences) or a monoclonal rat-anti mouse F4/80 (eBioscience, San Diego, CA). Isotype controls included PE-conjugated rat-IgG2a and nonimmune rat IgG, respectively. A preliminary step was included to block endogenous biotin (avidin/biotin blocking reagent, Vector Laboratories, Burlingame, CA) in the F4/80 staining procedure. Cell associated anti-F4/80 was detected using a biotinylated anti-rat IgG (Sigma-Aldrich, St. Louis, MO) followed by Alexafluor 660 streptavidin (Invitrogen Molecular Probes). The stained LF were postfixed with 0.5% paraformaldehyde. Epithelial cells were detected with monoclonal mouse-anti-cytokeratin 18 following fixation, permeabiliaztion, and blocking with Mouse-on-Mouse blocking reagent, as we had done for αSMA. The mouse anti-cytokeratin was detected using AlexaFluor 647- conjugated goat anti-mouse IgG. Flow cytometry was performed using the LSR II instrument. The proportions of cells staining positive for CD31, F4/80, or cytokeratin 18 were corrected for background staining.
Definition of Circumferential Deposits of Elastic Fibers and aSMA in the AERs
Tissues from the lungs of mice at postnatal days 4, 8, and 12 were obtained and processed as previously described. After removing the OCT and permeabilizing with 0.1% Triton X-100, the tissue slices were incubated with 2.5 μM eosin Y at 25°C for 1 hr (Mergens et al., 2007). After rinsing, the slices were imaged using the Zeiss 510 confocal microscope to visualize GFP and eosin Y (excitation 514, 550 long pass emission filter). Background staining with Eosin Y was subtracted and contiguous pixels demonstrating linear deposits of elastin were highlighted using uniform algorithms defined in IPLab. These included a background subtraction with a roll ball radius of 15 and a 3 × 3 pixel median filter. The same algorithm was applied to all images which enhanced visualization of elastic fibers. All of the AERs were visualized within a particular field, which was chosen at random. A superimposing line was drawn over collections of bright red pixels (elastic fibers) that appeared in the z-slice that best depicted the entry ring or from z-slices position one level above or below the slice at the entry ring. Entry rings were defined by collections of GFP+ cells and by their junction with alveolar ducts. The alveoli that were analyzed had a discrete beginning and ending within the tissue section, and needed to branch from the alveolar duct at an approximately 90-degree angle. The circumference of the AER was also traced. The number of pixels occupied by the tracing overlying elastic fibers was expressed relative to the pixels occupied by the circumference of the alveolar duct. This adjusted the length of elastic fibers for the size of the respective AER. The same tracing protocol was applied to images that had been stained for αSMA. The images were obtained from lungs of the same mice that were used for imaging elastic fibers, but not from precisely the same microscopic fields. Deposits of elastin and αSMA were studied in 60 to 75 AERs for each lung and changes in the circumferential length of elastic fibers was compared to that of αSMA at the three different ages.
Enumeration of Proliferating Alveolar Cells
Anti-Ki67 antibody was used to identify cells that were progressing through the cell cycle (not in G0), which we have designated as “proliferating.” After permeabilizing with Triton X-100, lung sections from mice at postnatal days 4, 8, and 12 were incubated for 90 min with a 1:100 dilution of anti-Ki67 (clone TEC-3, Dako, Carpinteria, CA). After washing, the tissues were incubated with 0.33 μM TO-PRO-3 (Invitrogen, Molecular Probes) and a 1:500 dilution of anti-rat-IgG-Alexafluor 546 for 1 hr at 25°C. After a final rinse, the tissue was mounted and examined using the Zeiss 510 confocal microscope with excitation wavelengths of 488, 543, and 633 to visualize GFP (PDGF-Rα-expressing nuclei), Ki-67 (nuclei of cells in S-phase), and TO-PRO-3 (all nuclei, Invitrogen Molecular Probes). Images were captured at 3 μm, z-stack intervals, using a 512 × 512 pixel density to allow a faster scan speed in order to avoid photobleaching of the TO-PRO-3. StereoInvestigator was used to analyze alveoli that had been transected so that at least 75% of the AER was visible in a single z-slice. A reference line was drawn around the AER, which demarcated the outer boundary of a cylinder of tissue that extended to the base of the alveolus. The optical fractionator probe was used to ascertain the volume density of all nuclei in the alveolar wall (TO-PRO-3), PDGF-Rα-GFP-expressing cells, and proliferating (Ki-67 containing nuclei). The volume of the cylinder served as the reference space and was normalized to the volume of the lung. Seventy-five alveoli from three lungs were examined at postnatal days 8 and 12, and from four lungs at postnatal day 4.
Localization of Lipid Droplets Within Alveolar Mesenchymal Cells
We first queried whether lipid-containing mesenchymal cells were more abundant at the AER or at the alveolar base. Because our prior studies showed that GFP containing cells were more abundant at the entry ring, and the BODIPY dye also served as a weaker counterstain for phospholipids, we could visualize the alveolar wall throughout its course from entry to base. The area of the AER was ascertained by tracing the outer and inner perimeters, and calculating the areal difference between the two tracings. The area of the alveolar base was ascertained as the area within the tracing, which outlined the column of alveolar airspace in the z-section closest to the base. A uniform segmentation algorithm was applied to both the entry ring and base to identify pixels with both red (vimentin) and blue (BODIPY 650/665) intensity that exceeded a particular threshold (magenta color). The threshold value was the same for both the entry ring and base. Segmentation was followed by an erosion to distinguish groups of pixels that were associated with lipid droplets from isolate pixels in the background. The pixel area of lipid droplets that were located at the entry ring was divided by the area of the entry ring, and a similar calculation was performed for the lipid droplets that were at the base. We avoided portions of z-stacks where there was a visible difference in the intensity of BODIPY staining at the AER compared to the base.
We also examined the pixel area of lipid droplets that were proximate to nuclei, which exhibited either a high or low level of PDGF-Rα gene expression (high or low intensity of GFP). Because the intensity of the BODIPY signal faded in the z-sections that were furthest from the laser beam, we examined a single slice that was 1/3 of the distance into each z-stack. All of the low and high-intensity GFP+ nuclei were examined if that particular slice was closest to the center of the nucleus. A uniformly sized circle (11.2 μm diameter) was placed around the centroid of the nucleus and lipid droplets that fell within the circle were identified. The pixel areas occupied by the droplets were ascertained for each nucleus that was examined. This allowed us to compare the area occupied by lipid droplets around either high or low intensity GFP+ cells within the same z-section. The mean lipid droplet pixel area for high PDGF-Rα expressing was calculated and subtracted from the area for both high and low level expressing cells. This reset the mean for the high expressing cells to zero, and provided a mean and variance of pixel area by which the low expressing cells exceeded that of the high expressing cells.
Is There a Temporal Correlation in the Abundance of PDGF-Rα-Expressing Alveolar Cells, aSMA and elastin?
We first examined the abundance of PDGF-Rα-expressing alveolar cells at postnatal (P) days 4, 8, and 12 (P-4, P-8, and P-12). A representative z-stack defining an alveolus at postnatal day 8 is shown in Fig. 1A. The AER is defined by the red αSMA in frame 1, whereas the base of the alveolus is defined by the continuous sheet of blue DiD stained cells in frame 5. The number of GFP+ alveolar cells increased from P-4 through P-12 when expressed relative to the volume of the entire lung parenchyma (Fig. 1B, left). However when expressed with respect to unit lung volume (mm3), a significant increase was observed in P-8 compared to P-4 lungs, but not when compared to P-12 (Fig. 1B, right). These data indicate that the abundance of PDGF-Rα-expressing cells increases as the alveolar septa form.
To determine whether PDGF-Rα-expression correlates with a myofibroblastic phenotype, we isolated a subpopulation of alveolar cells with characteristics of lung fibroblasts (LF). Approximately 85% of the isolated cells on postnatal day 12 were characterized as fibroblasts because endothelial cells, macrophages, and epithelial cells comprised 1.6% ± 0.3% (N = 3), 6.6% ± 0.3% (N = 3), and 8.1% ± 4.1% (N = 2), respectively. The isolated LF were stained for αSMA and separated into GFP+ (express PDGF-Rα) and GFP− (do not or only express low levels of PDGF-Rα) populations using flow cytometry. A significantly larger fraction of the GFP+ LF contained αSMA (Fig. 2, right side) compared to LF which did not express the GFP transgene (Fig. 2, left side). This indicates that PDGF-Rα expressing LF are more likely to contain αSMA, a distinguishing feature of MF, than are LF that are not expressing PDGF-Rα.
If PDGF-Rα-expression identifies a MF-phenotype, then PDGF-Rα-expression may persist as αSMA and elastin, which are produced by MF, accumulate in the AER. We analyzed collections of αSMA and elastic fibers in the AER. Exemplary AER at postnatal days 4, 8, and 12 are shown in Fig. 3A. The AER was identified as the opening of pockets of air that closed into a sheet of cells within the z-stack (alveolar base). The circumference of the AER was traced and a concentric line was traced over the immunoreactive αSMA filaments in the AER. This allowed us to assess the proportion of the AER circumference that was occupied by αSMA at P-4, -8, and -12. The proportion of the AER circumference that was occupied by elastic fibers was likewise determined using sections from the same mice that had been stained with eosin Y (white lines in Fig. 3A). Figure 3B shows that elastic fibers occupied a significantly lower proportion of the AER circumference at P-4 than at either P-8 or P-12. In contrast, αSMA occupied a similar proportion of the AER circumference at days P-4 and P-8, but decreased significantly by P-12. These findings indicate that PDGF-Rα expression persists throughout the period when αSMA and elastin accumulate in the AER, and therefore could participate in the accumulation. These data also show that in the AER, αSMA is more abundant relative to elastin at P-4 than it is at P-8 and P-12.
Is There a Spatial Correlation Between PDGF-Rα Expression and the Accumulation of αSMA?
We hypothesized that if PDGF-Rα expression correlates with a MF-phenotype, then PDGF-Rα expressing alveolar cells would be predominantly located in the AER where αSMA and elastin were most abundant. To assess whether PDGF-Rα-expressing alveolar cells are proximate to accumulating collections of αSMA, we evaluated the colocalization of the green (GFP, G) and red (cy3-αSMA) containing pixels (yellow, Y). We observed an age-related increase in the pixel area occupied by yellow relative to the pixel area occupied by green, within the area occupied by the AER (Fig. 4, left). This suggests that there is an age-related increase in αSMA in close proximity to GFP+-containing nuclei. We also calculated the area occupied by yellow pixels relative to the area of the AER (Fig. 4, right). This ratio also increased, which indicates that PDGF-Rα-expressing (GFP+) cells accumulated in the entry ring during the period from 4 to 12 postnatal days. Taken together with the data demonstrating that PDGF-Rα-expressing cells increase over this same period, these findings indicate that PDGF-Rα-expressing cells are accumulating in the same locale with αSMA. Albeit indirectly, these data further support the finding that PDGF-Rα-expressing cells, which accumulate in the AER, produce αSMA.
Do PDGF-Rα Expressing Progressively Colocalize with Accumulations of αSMA During Septal Elongation?
It is well established that septal elongation occurs at a maximal rate between postnatal days 4 and 12 (Burri, 2006). Therefore we compared the distribution of PDGF-Rα-expressing cells along an axis reaching from the AER to the base of the alveolus. We arbitrarily divided this axis into five segments (quintiles) so that we could compare data from saccular structures (terminal alveolar ducts or alveoli) at different ages (Figure 1A). At all three ages, the largest fraction of PDGF-Rα-expressing cells was found in the quintile located nearest the AER (quintile 1), ranging from approximately 37% to 48% at P-4 and P-12, respectively (Fig. 5). The majority of PDGF-Rα-expressing cells were found in the first two quintiles at all ages: 52%, 72%, and 68% at P-4, P-8, and P-12, respectively. The distribution at P-4 was unique in that the second most populated quintile was at the alveolar base, which contained the smallest fraction at P-8 and P-12. We analyzed the shape of the curves which fit a second-order polynomial distribution of the data at the three ages. The second moment of the polynomial varied significantly with age (one-way ANOVA on ranks, P < 0.05) indicating that the distribution of PDGF-Rα cells changed significantly after P-4, as a larger majority of the expressing cells were located at the AER. This could result from migration of PDGF-Rα-expressing cells from the base to the septum, proliferation of PDGF-Rα-expressing cells at the AER, or death of PDGF-Rα-expressing cells at the alveolar base. To more directly address the second possibility we evaluated alveolar cells that had exited G0 entered the proliferative portions of the cell cycle.
PDGF-Rα-Expressing Alveolar Cells Increase with Age Whereas Proliferating Alveolar Cells Decrease
PDGF-A stimulates mesenchymal cell proliferation, but PDGF-Rα-expression does not always predict cellular proliferative index (Karlsson et al., 1999, 2000). To examine whether PDGF-Rα-expression connoted a more proliferative phenotype, we determined the proliferative indices of all alveolar cells and of GFP+ expressing alveolar cells. A representative field from the lung at P-4 is shown in Fig. 6A and illustrates localization of Ki67 and GFP to the nucleus. The fractions of Ki67-staining cells relative to total alveolar cell (nuclear stain, Ki67+/nuclei) remained constant, whereas the fraction of Ki67-staining, PDGF-Rα-expressing cells (Ki67/GFP+) were maximal at P-4 and diminished with age (Fig. 6B). Therefore we observed an inverse correlation between the age-related changes in the fraction of proliferative PDGF-Rα-expressing cells and the increase in the number of PDGF-Rα expressing cells (Fig. 1B). These findings suggest that alveolar cells that express PDGF-Rα are not proliferating more rapidly than cells that do not express the receptor.
PDGF-Rα-Expression Inversely Correlates with Lipid Droplet Accumulation
Others have shown that the abundance of alveolar interstitial cells, which contain lipid droplets, decreases during the period of maximal septal elongation in mice and rats (Maksvytis et al., 1981; Awonusonu et al., 1999). This decrease coincides with the assumption of a more MF-like phenotype. We and others have shown that the lipid storage and MF phenotypes represent opposing poles of the spectrum of neonatal LF phenotype. A similar polarity has been observed in hepatic stellate cells are activated by PDGF-A, they store less retinyl ester and contain fewer lipid droplets (Moreira, 2007). Therefore we queried whether the abundance of lipid droplets varied along the axis extending from the AER to the base, and whether the distribution of droplets along this axis changed with age. We evaluated pixels that registered fluorescence from both BODIPY-650/665 and vimentin, to select for neutral lipids within mesenchymal cells. Figure 7A illustrates a representative field showing localization of blue BODIPY 650/655-stained lipid droplets with vimentin in some (*) but not other cells (τ). We found that the pixel density of lipid droplets in mesenchymal cells was approximately 3.6 and 2.1 fold higher at the base on P-4 and P-8, which significantly differed from the distribution at P-12 when there was no difference between the pixel density of lipid droplets at the base and entry ring (Fig. 7B). This suggests that lipid droplets are predominantly located at and preferentially disappear from the base as the septa elongate. We also queried whether lipid droplet accumulation varied with the level of PDGF-Rα gene expression. To assess this, we analyzed the pixel area occupied by lipid droplets within a fixed radius around alveolar cells with high versus low levels of PDGF-Rα (high versus low intensity of GFP-emission). We compared lipid droplets around nuclei with high and low level GFP-signal, in one z-slice. The mean pixel area that surrounded cells with the brightest GFP-emission for a particular slice was subtracted from the individual pixel areas for all high and low level GFP-expressing nuclei in that slice. This enabled us to calculate a mean pixel area for the population of either high or low expressing nuclei in each slice that was analyzed, and to normalize for variance in the BODIPY signal intensity in different stacks (which differed somewhat in their total height). We found that the pixel area around cells with low level PDGF-Rα-expression significantly exceeded the area around cells with high level expression at P-4 and P-8, but not at P-12 (Fig. 8). When combined (Figs. 7 and 8), these data indicate that when lipid droplets are preferentially located at the alveolar base, they are also preferentially located around cells with a lower level of PDGF-Rα expression. This suggests that lipid droplets are primarily lost from alveolar cells with the lowest level of PDGF-Rα expression by day 12.
The temporal course and stereology of alveolar septal formation have been extensively characterized using thin sections embedded in paraffin or plastic (Thibeault et al., 2003; Burri and Schittny, 2004). In contrast, relatively few studies have been conducted using optical slices obtained from confocal microscopy (Oldmixon et al., 2001). We opted to use confocal microscopy for several reasons. First, we desired to preserve the fluorescent emission of GFP and avoid autofluorescence induced by plastic embedding media. Second, because we relied on immunohistochemical identification of αSMA, Ki67, and vimentin we wanted to avoid alterations in antigens that may result from paraffin embedding or antigen retrieval. Third, dehydration, paraffin embedding, and dewaxing would have prohibited the detection of neutral lipid droplets. And fourth, optical sectioning is ideally suited to the optical fractionator probe, which allowed us to count nuclei using an unbiased stereological approach. Although it was advantageous for our objectives, confocal microscopy does have limitations. First, the stage of the Zeiss 510 microscope was only powered by a motor in the z-axis, so fields could not be randomly selected in an automated fashion. Therefore we introduced randomness into the selection process by choosing areas using lower power so that the particular cellular features that we were studying were not sufficiently magnified to influence the decision. In all cases, fields that contained large airways or blood vessels, pleura, or were incompletely expanded were avoided. We chose the sizes of the pinhole and optical slice to limit optical distortion that would render it difficult to determine which section the cell nucleus primarily resided in. Finally, we used uniform criteria for selecting alveoli for analysis and avoided alveoli that arose from alveolar ducts at an obtuse angle.
Several priorities were considered in our choice of stereological methods. First, we limited the use of staining intensity to quantify proteins or structures. Antibodies for αSMA, vimentin, and Ki67 were used to identify structures or cells: the AER, mesenchymal cells, and cycling cells, respectively. The AER was used to demarcate the junction with the alveolar duct, vimentin was used to identify lipid droplets that were associated with mesenchymal cells, and Ki67 was used to enumerate proliferating cells. To further limit the effects of variations in staining intensity, we always concurrently processed tissues from mice at all three ages. Although we did not use intensity to evaluate αSMA, Ki67, or vimentin, this parameter was involved in the quantification of BODIPY-650/665 in neutral lipid droplets. In this instance, the intensity of the dye in lipid droplets was combined with uniform criteria for segmentation that allowed us to consistently measure the area of pixels meeting the criteria. Finally, we avoided counting structures in two dimensions by using the optical fractionator. This enabled us to avoid errors from counting structures with a centroid outside the plane of the section and allowed us to express the data relative to the volume of tissue.
Studies by Böstrom et al. and Lindahl et al. compared PDGF-A null and wild-type mice (Bostrom et al., 1996, 2002; Lindahl et al., 1997). They demonstrated that PDGF-Rα mRNA localized to blood vessels, airways, and scattered cells in the lung parenchyma after birth and the abundance of mRNA diminished after P-7. Alveolar septal elastic fiber formation was markedly diminished in the PDGF-A null mouse, which also exhibited a striking absence of PDGF-Rα in the postnatal alveolar sacs. However, these studies did not demonstrate a temporal or spatial association between PDGF-Rα-expressing cells and tropoelastin gene expression. The abundance of PDGF-Rα mRNA diminished prior to the decrease in tropoelastin mRNA. Whereas tropoelastin mRNA appeared most abundant at the septal buds or tips, PDGF-Rα mRNA localized in scattered cells that were removed from the vicinity of the septal buds and tips. Furthermore, αSMA was only observed in the airways and vasculature during the second postnatal week, and became most abundant after the maximal period of tropoelastin synthesis. These investigators observed a normal complement of PDGF-Rα containing cells in the PDGF-A null mice, which persisted until birth in the distal tubules and primitive sacs, but an absence of these cells by P-7. From these data, the authors reasoned that PDGF-A may be required for the “spreading” of PDGF-Rα-expressing cells into the distal portion of the alveolar septum.
Although our studies corroborated some of the findings of Böstrom et al. and Lindahl et al., we also observed important differences. Like these investigators, we observed that the distribution of PDGF-Rα-expressing cells shifted (“spread”) more distally from the alveolar base to the AER between postnatal days 4 and 8. However, unlike Böstrom et al. and Lindahl et al., we did not observe a decrease in PDGF-Rα-expressing cells by P-8, and instead the abundance of these cells was sustained at least until P-12 (Fig. 1). We found that the most prominent collections of PDGF-Rα-expressing cells, αSMA, and elastic fibers all localized to the AER, whereas the studies by Böstrom et al. and Lindahl et al. did not exhibit spatial or temporal colocalization of these components. We attribute these disparities to differences in the histological approaches that were used. Böstrom et al. and Lindahl et al. relied on in situ hybridization and immunohistochemistry using thin sections of tissue and quantitative stereology was not performed. Using GFP to localize PDGF-Rα expression allowed us to more precisely identify and quantify expressing cells. Another advantage of our approach was that it allowed us to localize proteins or expression patterns to identical or contiguous pixels, rather than general regions in sections that were obtained from somewhat different portions of the lung. The greater precision of our methods supports the significance of our finding that PDGF-Rα-expressing cells colocalize both spatially and temporally with αSMA. We also observed that newly isolated PDGF-Rα-expressing fibroblasts are more likely to contain αSMA than fibroblasts that are not expressing PDGF-Rα (Fig. 2). When combined, these findings indicate that cells, which are expressing the pdgf-Rα gene, concurrently produce αSMA and that they contribute to the denser accumulation of αSMA at the AERs. We also observed that a larger proportion AER circumference was occupied by αSMA than by elastic fibers at P-4 (Fig. 3). However by P-8 this disparity had disappeared and by P-12 αSMA occupied a smaller proportion of the circumference than at P-8. Because mechanical tension at the AER is required to prevent collapse during expiration, αSMA may play a more critical mechanical function at P-4 than at P-12 when elastic fibers are more abundant in the AER. This would be consistent with recent observations that tension stimulates αSMA gene expression (Zhao et al., 2007). As elastin becomes more abundant in the AER, the tension exerted on αSMA filaments would diminish and one would expect that αSMA may be less abundant, which is what we observed at P-12.
Although our studies have more precisely mapped the temporal and spatial locations of PDGF-Rα-expressing alveolar cells, they do not demonstrate that PDGF-A was directing these changes. However observations by others suggest that PDGF-A is a significant contributor. First in its absence, PDGF-Rα-expressing cells fail to populate the locations where we observed them (Bostrom et al., 2002). Second, although PDGF-C also requires PDGF-Rα for signaling, and PDGF-C is expressed in the lung in utero, abnormalities were not observed in the lungs of PDGF-C null mice at birth (Ding et al., 2004). These mice could not be studied postnatally because they are unable to suckle and die shortly after birth. Therefore PDGF-A is also likely to be the relevant PDGF-Rα ligand in our studies. Other soluble factors and extracellular components, which do not directly interact with PDGF-Rα, also likely impact the persistence and migration of the alveolar MF.
Our primary purpose for characterizing the abundance and location of PDGF-Rα-expressing alveolar cells was to map the temporal and spatial fate of the alveolar MF. Therefore the PDGF-Rα-GFP construct enabled us to address this intent. Others have used hemizygous PDGF-Rα gene insertions to map the temporal and spatial fate of the neural crest and pulmonary mesenchymal cells (Klinghoffer et al., 2002; Mandeville et al., 2006). Zhang et al. demonstrated that the PDGF-Rα-LacZ transgene was expressed in the mesenchyme of the lung parenchyma during gestation and through postnatal day 7 (Zhang et al., 1998). The abundance of the LacZ gene product diminished after P-7 and was absent by P-20. More recently PDGF-Rα-GFP marker was used to show that Hox 5a gene deletion resulted in diminished migration and elastic fiber production by embryonic lung MF (Mandeville et al., 2006). Therefore a precedent exists that PDGF-Rα-expressing alveolar cells demonstrate a MF-like phenotype.
During the first 3 weeks of postnatal life in rats and mice, two LF subpopulations can be distinguished based on their abundance of neutral lipid droplets namely the lipid and nonlipid interstitial fibroblasts (LIF and NLIF). LIF predominantly found at the base of the septa (Burri, 1974) have a lower proliferative index than NLIF at P-4, and both LIF and NLIF proliferation diminishes significantly by P-10. The abundance of LIF decreases relative to that of NLIF between P-4 and P-12 and there is a precipitous decline in LIF after P-15. This decline results from apoptosis of the LIF and 51% of this subpopulation exhibits DNA fragmentation between P-15 and P-17. The LIF subpopulation has been further distinguished as having either a higher (LIF+) or lower (LIF−) abundance of lipid droplets. The LIF− retain insulin like growth factor 1-receptor mRNA and function and are more likely to avoid apoptosis (Srinivasan et al., 2002). Our findings corroborate those reported by Burri and by Kaplan and Brody demonstrating that LIF are more abundant at the base of the secondary septa. We have made the novel observation that alveolar cells with a lower intensity of GFP-fluorescence exhibit a greater areal density of surrounding lipid droplets. Because we analyzed pixel density of lipid droplets and the intensity of GFP in the same z-slice, it is unlikely that differences in excitation beam penetration or quenching of the emitted light account for our observation. Therefore differences in GFP-signal intensity most likely result from differences in the abundance of the GFP-protein, which is directly proportional to PDGF-Rα promoter activity. Taken together these findings indicate that lipid droplet accumulation is most abundant in cells with lower PDGF-Rα gene expression. This is consistent with observations that others have made using hepatic stellate cells (Breitkopf et al., 2005).
In conclusion, we have shown that PDGF-Rα-expressing alveolar cells are preferentially located at the AER and produce αSMA. The proportion of alveolar PDGF-Rα-expressing cells that localize to the AER increases between postnatal days 4 and 12, whereas the proportion located at the alveolar base decreases. This is consistent with relocalization of PDGF-Rα-expressing cells from the alveolar base to the AER. The accumulation of αSMA in the AER occurs earlier after birth than the accumulation of elastin. Therefore, αSMA may serve an important mechanical function: to stabilize the AER during expiration, until elastic fibers accumulate. The level of PDGF-Rα expression is inversely correlated with lipid droplet accumulation which preferentially occurs at the alveolar base. Taken together, these findings indicate that PDGF-Rα-expressing alveolar cells are more likely to have myofibroblastic rather than lipid storage characteristics. Whereas the myofibroblast phenotype may be essential during secondary septation, it may be undesirable in the adult lung that is undergoing a fibrotic response. Strategies that modulate PDGF-Rα-expressing cells toward a lipid-storage phenotype may alter the outcome of fibrotic lung diseases (Rehan et al., 2006).


text:
I know that is the next step.
I am so tired of paying for myself and paying for all these people who are illegally here. It is all for no good reason other than to bring us all to our financial knees.
I am looking at a bill I got from the hospital for my sons surgery. Not only that, but I also got a request from the hospital for a DONATION for people unable to pay their bills.
What a joke, and in poor taste.
Yep. It's all that compassionate convervatism.

domain: medical


text:
Is Redundancy Pay Taxable
The good news is that the first £30,000 of a redundancy payment is tax free.
Everything over £30,000 is then taxed at your highest rate and will usually be deducted under Pay As You Earn. You will pay tax on your wages as well as pay in lieu of holiday and any bonus payments.
Do I pay tax on my wages?
Yes, you will still pay tax on any of your unpaid wages as normal, but not on the separate redundancy payment. You will also pay tax on any bonus you receive and on your occupational pension.
Will I still have to pay National Insurance on my Redundancy Pay?
Yes, you will still pay your national insurance on your unpaid wages and any bonus payments but not on your occupational pension. You will not pay any on your redundancy payment

domain: economics


text:
An Instagram star is turning heads for her startling resemblance to singing superstar Beyonce.
>> Read more trending news
Brittany Williams shares flawless good looks with Beyonce, and her resemblance to the star causes quite a commotion when she hits the town.
"I get approached all the time, whether it be on planes, at the airport or while attending events," Williams told the Daily Mail.
“I’ve also been chased, had pictures taken of me without my consent and pranks done without me knowing,” she said.
“A group of women once chased me and my friend to our car and began singing ‘Single Ladies,’ beating my friend’s car with the heels of their shoes until we rolled down the window and took a picture with them.”
Her good looks aren’t the only thing Williams shares with the mother of Blue Ivy, Sir and Rumi. She also sings, dances and writes.
Williams admits that her resemblance to Beyonce used to bother her, but she has learned to embrace her doppelganger status. The Detroit, Michigan, native is also learning to deal with the attention, which she admits can be overwhelming.
“I’m just a cool, stylish, smart, witty girl with vision, confidence and ambition. I’m still dreaming that God will allow my visions to come to fruition,” she confidently told the Daily Mail.

domain: celebrity


text:
Gift certificate for one of our one hour group riding lessons (english or western style).
* Please note in order to ride in a group lesson the reciepent must have prior riding experience and be able to ride on their own. If the do not please purchase the private lesson option so the instructor will be able to work with the student one on one.
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domain: hobby


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The 5th India- Myanmar Bilateral Meeting on Drug Control Cooperation between the Narcotics Control Bureau, India and the Central Committee on Drug Abuse Control, Myanmar was held virtually yesterday. The Indian delegation was led by the Director General Narcotics Control Bureau, Rakesh Asthana and the Myanmar delegation was led by the Commander of the Drug Enforcement Division cum Joint Secretary of the Central Committee for Drug Abuse Control, Pol. Brig. Gen. Win Naing.
Mr Asthana highlighted issues particularly the trafficking of Heroin and Amphetamine Type Stimulants ATS in the country. Narcotics Control Bureau has remained committed towards strengthening the existing mechanism of sharing information and assistance with Myanmar, for combating the drug menace in the region.
The Commander of Drug Enforcement Division, Myanmar, Pol. Brig. Gen. Win Naing urged India to develop frequent information exchange on trafficking of drugs and precursor smuggling activities at every level. The Commander Drug Enforcement Division complimented the Government of India and the Narcotics Control Bureau for their continuous efforts to combat the growing threat of the drug menace.
Both countries agreed on the exchange of intelligence information in a timely manner to conduct follow-up investigations in drug seizure cases, new psychotropic substances and their precursors. The 6th India – Myanmar Bilateral Meeting on Drug Control Cooperation will be held in India next year.
Support Ethical Journalism. Support The Dispatch
The Dispatch is a sincere effort in ethical journalism. Truth, Accuracy, Independence, Fairness, Impartiality, Humanity and Accountability are key elements of our editorial policy. But we are still not able to generate great stories, because we don’t have adequate resources. As more and more media falls into corporate and political control, informed citizens across the world are funding independent journalism initiatives. Here is your chance to support your local media startup and help independent journalism survive. Click the link below to make a payment of your choice and be a stakeholder in public spirited journalism

domain: news


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Hangovers can be a real pain, and if you’ve ever experienced one, you know how difficult it can be to shake off the unpleasant symptoms. Nausea, headaches, and fatigue are just a few of the side effects of consuming too much alcohol. However, did you know that taking a cold water shower could help alleviate some of these symptoms? In this article, we’ll explore the benefits of a cold shower for a hangover and how it can help you get rid of it.
What is a hangover?
A hangover is a collection of unpleasant symptoms that occur after drinking too much alcohol. It is a common condition that affects many people, and the severity of the symptoms can vary from person to person. The most common symptoms include headaches, nausea, dizziness, fatigue, and sensitivity to light and sound.
What causes a hangover?
Alcohol is a diuretic, which means it causes the body to lose water through increased urine production. This can lead to dehydration, which is one of the main causes of a hangover. Alcohol also irritates the lining of the stomach, which can cause nausea and vomiting. Additionally, alcohol can interfere with the body’s ability to produce glucose, which can lead to fatigue and weakness.
How Cold Shower Is Helpful For Hangover?
Taking a cold shower can help alleviate some of the symptoms of a hangover. Here are some of the ways a cold shower can help:
- It can help reduce inflammation
Alcohol consumption can cause inflammation in the body, which can contribute to symptoms like headaches and joint pain. Taking a cold shower can help reduce inflammation by constricting blood vessels and reducing blood flow to affected areas.
- It can help with dehydration
As mentioned earlier, dehydration is one of the main causes of a hangover. Taking a cold shower can help rehydrate the body by increasing blood flow to the skin and reducing body temperature. This can help improve circulation and reduce inflammation, which can in turn help with symptoms like headaches and fatigue.
- It can help improve mood
Hangovers can often leave us feeling grumpy and irritable. Taking a cold shower can help improve mood by releasing endorphins, which are natural painkillers that can help reduce anxiety and improve mood.
- It can help with nausea
Nausea is a common symptom of a hangover. Taking a cold shower can help reduce nausea by constricting blood vessels and reducing inflammation in the body. This can help improve circulation and reduce inflammation, which can in turn help with symptoms like headaches and fatigue.
- It can help with alertness
Hangovers can often leave us feeling groggy and unfocused. Taking a cold shower can help improve alertness by increasing blood flow to the brain and reducing body temperature. This can help improve concentration and focus, which can help with productivity throughout the day.
Hot or Cold Shower For Hangover
While both hot and cold showers have their benefits for hangover relief, a cold shower is generally more effective. Cold water helps to constrict blood vessels and reduce inflammation, which can alleviate headaches and other pains caused by a hangover. It can also help increase alertness and provide a burst of energy, which can combat feelings of fatigue and sluggishness.
On the other hand, hot water can help relax muscles and ease tension, which can be helpful for hangover-related muscle soreness. However, it can also increase blood flow and potentially exacerbate headaches or nausea.
Ultimately, the decision between hot or cold shower for hangover relief depends on personal preference and how your body responds. If you find that a cold shower is too uncomfortable, you can try alternating between hot and cold water or starting with a warm shower and gradually decreasing the temperature to achieve the desired effects. It’s essential to listen to your body and make sure you don’t expose yourself to water that is too hot or too cold, as this can be harmful to your health.
Tips for Taking a Cold Shower for Hangover
A cold shower can be effective in reducing hangover symptoms because it helps to constrict blood vessels and reduce inflammation, which can help to relieve headaches and other pains. Additionally, the shock of the cold water can help to increase alertness and provide a burst of energy, which can help to combat feelings of fatigue and sluggishness.
To take a cold shower for hangover relief, start by adjusting the temperature of the water to the coldest setting. If you find the water to be too cold, you can gradually increase the temperature until you reach a comfortable level. It’s important to note that you should never expose your body to water that is too cold, as this can cause hypothermia or other health problems.
Once you’re in the shower, spend a few minutes under the cold water, focusing on the areas where you’re experiencing pain or discomfort. You can also try alternating between hot and cold water to stimulate blood flow and increase circulation. Finish your shower with a few minutes of warm water to help relax your muscles and ease any tension.
In addition to taking a cold shower, there are several other things you can do to help alleviate hangover symptoms. Drinking plenty of water to rehydrate your body, getting enough rest to allow your body to recover, and eating a healthy meal to replenish nutrients are all important steps in overcoming a hangover. You can also try taking over-the-counter pain relievers, such as ibuprofen or acetaminophen, to help alleviate headaches or other pains.
Taking a cold shower can be an effective and natural way to alleviate hangover symptoms. By constricting blood vessels and reducing inflammation, a cold shower can help relieve headaches and other pains. Additionally, the shock of the cold water can help increase alertness and provide a burst of energy, which can help combat feelings of fatigue and sluggishness. Other remedies such as drinking plenty of water, getting enough rest, eating a healthy meal, and taking over-the-counter pain relievers can also help alleviate hangover symptoms. So, the next time you’re feeling the effects of a night of drinking, consider taking a cold shower to help you feel better.
Are cold showers good for hangovers.
Will a cold shower help a hangover.


Populist anger should be appeased by the Fed’s emphasis on higher-than-usual capital requirements, an attempt to make sure banks survive the recession, especially if it lasts longer than expected. And short sellers will be reluctant to raid banks because there is no guarantee the Fed will punish those that fail this test.
“All they’re trying to do is stall and delay,” Cramer said of Bernanke and Geithner.
The one negative out of today’s news? Neither Goldman Sachs nor JPMorgan Chase will be allowed to return the Troubled Assets Relief Program money they borrowed for the time being. That, Cramer said, ruins their potential competitive advantage.
Cramer's charitable trust owns Goldman Sachs and JPMorgan Chase.
Questions for Cramer? email@example.com
Questions, comments, suggestions for the Mad Money website? firstname.lastname@example.org


text:
Minos electronic Armchair
The Ontario chair can control two motors independently reclining back and leg elevation ...
The Invacare Porto comes with a new design, with different functions and options of upholstery, specially designed to fit into all environments and meet the needs delos users.
- Online only
The lifting and relaxing chairs have a fully electronic operation.
The chair Faro exists in version 1 motor that can lift the footrest and recline the seatback in a single simultaneous movement.
The Douro sofa offers additional comfort thanks to the continuity between the seat and the footrest.

domain: design


Oct 16, · CA cable drivers and Windows 8 (x64) Hi, Are the CA cable drivers compatible with Windows 8 available? The ones that I have on a CD worked with Windows XP, but I don't think they'll be good for Windows 8. In the Windows Control Panel» Software, what is the version of your Nokia Connectivity Cable Driver: ? In the Windows. The Connectivity Adapter Cable DKU-5 is compatible with phones with a Pop-Port™ interface connector, for example, the Nokia mobile phone. Please check the compatibility from your phon e's accessory guide. The Connectivity Adapter Cable driver software is c . Jun 25, · Install the driver to establish a cable connection between your compatible Nokia phone and a compatible PC. Important for Windows or XP or Vista users: Drivers for the Nokia DKE-2, DKU-2, CA, CA, CA, and CA cables are now integrated into PC Suite.
Driver for connectivity adapter cable ca-42Where can I find the ORIGINAL drivers of cables? How come the connector does not seems to fit the phone / too tight? The ORIGINAL CA / DKU-5 cables are quite expensive and hard to get elsewhere but Nokia itself. The Nokia Connectivity Adapter Cable CA is for you if you want a USB cable This software package does include a driver but not for this cable (only for the. Nokia CA Connectivity Adapter Cable for Nokia Phones; ›; Customer reviews As a matter of fact you must use the driver CD, if you let Windows 7 find the. and it will not read the software disc for the connectivity adapter cable. The genuine Nokia CA cable driver does not match other IDs. Genuine USB data lead CA for access of data between your PC and Nokia phone. - Nokia CA Connectivity Adapter Cable. If I connect CA cable to USB port without any phone connected Depending on what version of windows you are running, the drivers Serial is very different than jtag. you will us the ca as a serial connection, not a jtag connection. that got their power from the phone connector instead of the USB. Brand New Nokia CA Connectivity Adapter Cable USB Data Cable CA DKU-5 For NOKIA i .. To use it on Win10 computer, you should install custom drivers first then you should use OXYGEN PHONE. Install Java applications/games from the PC to the phone; Send text messages and edit the phone book and message settings of your phone. Edit and backup. The drivers for Nokia cables. If you have a Nokia This driver is compatible with the following Nokia data cables: Nokia Connectivity Adapter Cable CA
See This Video: Driver for connectivity adapter cable ca-42
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I protest against it.


text:
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LANDSCAPING SERVICES ▾
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EARTHMOVING & CIVIL CONSTRUCTION
BOBCAT AND SKID STEER HIRE
Earthscape Industries provides expert pool landscaping solutions in Sydney, the Hills District, the Hunter Valley, the Southern Highlands and beyond. Swimming pools are a popular choice for Sydney homes, where the summers are scorching hot. A well-designed swimming pool is an excellent investment that can add value to your property and provide great home entertainment. Earthscape Industries specialise in pool landscaping services across Sydney and surrounding areas. We specialise in the design and construction of in-ground swimming pools, designed to elevate the appearance of your property. Whether you are looking to build a lap pool for exercise, a pool for the whole family, a spa or a plunge pool, Earthscape Industries can help.
Trees, shrubs, hedges and plants will provide a beautiful finish to your newly installed pool. They can provide shade and added privacy, as well as an aesthetic look. Earthscape Industries specialise in delivering beautiful, quality pool landscaping finishes incorporating a combination of natural and manufactured products. We strive to create beautiful living spaces that blend seamlessly into your property and the surrounding environment.
Our team of highly experienced horticultural specialists can assist you to choose shrubbery and plants that will thrive in the surrounding environment and elevate the look of your swimming pool. Earthscape Industries offer horticultural services and supply plants at a competitive rate. From conception to construction, we will work alongside you to provide beautiful pool landscaping finishes.
Earthscape Industries can assist you with any landscaping or earthworks project. From conception to construction we strive to deliver beautiful pool landscaping finishes. We proudly service Sydney, the Hills district, the Hawkesbury, the Blue Mountains, Galston and beyond. To discuss our pool landscaping services and receive a free quote, give us a call today. Alternatively, you can use our enquiry form and our specialist team will contact you.
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text:
I like to eat. In fact, I pretty much work out to support my eating habit. However, over the years I have learned a few tricks to keep my enjoyment from becoming overindulgence. One of those tricks is finding food substitutions that taste as good or better than the things I think I really want.
For example, I grew up a huge fan of Thousand Island salad dressing. A salad just wasn’t a salad without (lots of) it. When I finally decided I needed to lose weight I knew that drowning my salad in Thousand Island wasn’t probably the best way to go about doing so. I weaned myself off of it gradually until I no longer missed it. Do I still crave Thousand Island’s creamy goodness? Yes. But that’s mostly because I’m conditioned to want it. Once I get over myself and simply try something else (sometimes through sheer force of will) I find I enjoy my salad just as much. What I now use instead is a super flavorful Fat Free Balsamic Vinaigrette by Maple Grove Farms of Vermont. It’s really tasty so a little goes a long way and 2 Tbsp only has 5 calories. FIVE! I’ve never seen another salad dressing with so few. Even better, I know what all the ingredients are: Water, balsamic vinegar, cider vinegar, dehydrated garlic, salt, spices, cane syrup, and xanthan gum. (Ok, I don’t really know what xanthan gum is but you get what I mean.) A quick glance at the other balsamic dressing that’s been languishing in my fridge yields ingredients like potassium sorbate, propylene glycol, and calcium disodium EDTA. [Side note: I was a biology major and I distinctly recall measuring out EDTA for some gnarly chemical experiments. Oh, and it’s used as a detergent. YUM!]
I’d also highly recommend Fox Hollow Farm’s Fox Mustard. It’s a spicy sweet mustard that is awesome on sandwiches or as a dip for pretzels. I haven’t yet used it as a marinade as they suggest on the jar but I’m sure that would be wonderful too. It’s got 15 calories per teaspoon but packs enough kick that you don’t need more than that. And how’s this for ingredients: brown sugar, canola oil, balsamic vinegar, mustard flour, water, molasses, fresh garlic, and spices. That’s it. No additives or preservatives. Lots of taste.
If you have products you love – preferably natural ones that aren’t full of “-oses” and “-ates” and other things that can’t be pronounced – I’d love to hear about them. Bon appetit!

domain: food


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Lost Abbey Bottling New Blend
So today, we’re bottling Deliverance, and let me tell you, it’s one HEAVY beer.
What is Deliverance and how does one bottle it? Well, it’s a blend of Bourbon Barrel Aged Serpent’s Stout and Brandy Barrel Aged Angel’s Share. You see in Bourbon Barrel Aged Serpent’s Stout we have captured the essence of one very evil beer (the Devil’s brew if you will). Blacker than the strongest plague and hotter than Hades itself, there’s fire in this beer- a veritable cauldron of Bourbon infused beer. Left to its own devices, this would be one sinister liquid.
Yet, we have tempered this odious darkness with heaven sent Saintly created Angel’s Share. And together, these two beers will work in harmony to deliver us on that day.

domain: artistic


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Regardless of how you feel about EDM, Colorado is becoming one of the hotspots for developing new artists. Two such artists will be taking the Mesa Theater Main Stage this Saturday (Jan. 26). The Digital Connection (pictured) and Joman make the trek over the hills from Denver to test our soundsystems in what should be a fun night.
Both are no stranger to big stages. The Digital Connection played Snowball 2012, Sonic Bloom 2011-12, and recently a NYE set with Amon Tobin. Joman played Global Dance Festival last year. Check out The Digital Connection's chill dubstep remix of Deadmau5 and Kaskade's "I Remember." It's quality. For those 21 and up the night gets even better. There's no cover! For those 18 and up the cover is a reasonable $8.
On a side note Miss Emily's CD release party will still take place at the Mesa Theater. Her event starts at 7 p.m. while the first DJ wont start until 10 p.m.
The Coachella 2013 lineup is finally here and finally official. There are a couple little surprises here and a couple things worth paying attention to for the rest of the year. Here is my knee jerk, initial reaction.
First the good. With Phoenix and Vampire Weekend playing that means only one thing: they are armed with new music. It will be fun to follow their trajectory this year as both band's previous albums were really stinking good. Can they, especially Phoenix, live up to the hype? As a fan of both bands I hope so.
Dinosaur Jr. seems to have new life so their set will be interesting. I'm also curious what the crowds will be like for chart toppers Of Monsters and Men and the Lumineers. Former Smiths guitaris Johnny Marr could be on double duty as he will be playing solo and his former band Modest Mouse both play on Friday. Might they collaborate again?
Now the surprises. Who's excited for the English quartet The Stone Roses? The band hasn't really been active since 1996 but I guess the promised world tour and new album is finally happening. What about the Postal Service? Ben Gibbard doesn't play festivals with his main and more popular band Death Cab for Cutie but does with a side project that only released one album back in 2003? Curious. I'm also surprised by the lack of a big name DJs to blow the roof off the legendary Sahara Tent Stage. Last year they had Swedish House Mafia, Avicii, David Guetta, Martin Solveig, Afrojack, and Kaskade. That's massive. Hardwell and Bassnectar are great but hardly matchup to last year.
As usual the lineup is pretty stacked so if you are headed out to the desert there will once again be no shortage of great options.
The other day my friend and bandmate was talking about this crazy Swedish movie "Sound of Noise." The movie is about music hating and tone-deaf cop named Amadeus, and his pursuit of a group of guerilla percussionists "terrorizing" the city with their public performances. For the percussionists everything is an instrument. Coins, bulldozers, jack hammers, a car engine, stamps, an operating room, blenders, cabinet doors, a tooth brush, etc...
I can't speak for the quality of the entire movie but we checked out several performances and they are extremely creative. The music pieces are certainly avant-garde but all fans of music will appreciate the fact that literally anything can be turned into an instrument. Here is a peek at the group performing a piece in a kitchen. Enjoy.
It's a big week for music here in Grand Junction. So far I count at least four different shows worth checking out this Friday and Saturday and none of them are any bigger than the Reverend Horton Heat with Shotgun Hodown and Reno Divorce (Check out my full show preview in this Friday's Out and About).
If you are a gear head like me then you will get a kick out of this video. Want to get that signature Rev sound? Well it's going to cost you. Here is Jim Heath talking about the gear he uses on stage including his custom Gretsch hollowbody and his Gretsch Executive amp. Just between the two, you are looking at about $5,000 and that's not including the effect pedal set you would need to add.
The sound is a little spotty in this video but I think it's always cool to hear from musicians about what they look for in gear and how much energy they put in to get that perfect sound.

domain: entertainment


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Check out our new audio content!
Real estate investors take note of Realty411‘s upcoming financial webinar with Universal Commercial Capital scheduled for August 1st at 11 AM PST.
During this informative and educational online session, the team from UCC, Eric Tran, Ivy Baca and Milica Krstic, will discuss the current state of the financial market so investors can learn about their financial options.
With this unprecedented economic time amidst a global pandemic, it’s crucial for investors to be aware of the dynamic that affect private lending.
“Why Your Fix & Flip Deal is Turned Down by Lenders?” will provide answers to this, as well as many other important financial questions.
Organized by Realty411‘s publisher, Linda Pliagas, this special webinar is one that investors of all experience levels simply cannot afford to miss.
While the current economic situation may seem bleak, historically, it’s been proven that life-changing generational wealth is created precisely during times of financial turmoil.
“We are living in an unsettling time, one filled with fear and misinformation, which causing even more havoc on our markets.” Pliagas says, adding, “As investors we need to get rid of the noise and zero in on facts, which is why we are hosting this special webinar.”
With nearly 23 million renting families at risk for losing their homes by this fall, according to a recent study from the COVID-19 Eviction Defense Project, real estate investors will undoubtedly be affected by this economic tidal wave.
Join us and acquire the insight to correctly position yourself and your family’s real estate portfolio for long-term safety and continued growth.
Aug 1, 2020 11:00 AM PST
Webinar ID 859 7270 7741
To Join the Webinar:
Join from a PC, Mac, iPad, iPhone or Android device:
Please click this URL to join: https://us02web.zoom.us/webinar/register/WN_ErCBe8KASfuH-wbmKSBYLg


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Though long retired, Robert "Doc" Morgan remains a vital piece of Houston's jazz history, with ties to its present and future as well as its past. He grew up in Houston, the son of a saxophonist, and became an educator at Sam Houston State University and the High School for the Performing and Visual Arts, sending scores of performers - pianists Jason Moran and Robert Glasper among them - on to successful careers before he retired in 1999.
Sitting at the Montrose coffee shop Blacksmith, Morgan looks across the street at the restaurant El Real. "Ella Fitzgerald played five nights there back when it was a theater," he says.
His recall of jazz happenings in the city remains sharp.
Although Morgan receives unfettered reverence from his former students, many of the jazz artists who passed through HSPVA's storied jazz program tell tales about drawing his ire. Often the mistakes described aren't of a technical or instrumental nature but rather failing to show for a session or a performance or showing up unprepared. Each of these musicians, who have gone on to great things, speaks of those moments as transformative: The moment they began practicing a personal discipline to accompany their great musical talents.
Chris Walker - a bassist, composer and singer, who also works as bandleader for Al Jarreau - recalls one session that turned things around.
When: 6:30 p.m. Saturday
Where: Ensemble Theatre, 3535 Main
Tickets: $25-$100; docfestjazz.org
"I simply hadn't prepared," Walker says. "Doc comes in and starts counting it off, and David Craig, another bassist, and I hadn't prepared. Doc ripped us right there. He told us to go home and come back tomorrow."
Earlier this year, drummer Kendrick Scott - one of the top jazz drummers working today - played a hometown show. During a Q&A session with Morgan, he flipped the roles of interviewer and subject. Scott asked Morgan if the teacher recalled taking a David Sanborn composition away from Scott and giving it to another drummer for a performance.
"I wasn't really prepared for his question," Morgan says, laughing. "We're there on stage, in front of this crowd, with his mom in the audience. So, for a moment, it was a little bit uncomfortable. And he said, 'I want to thank you. I decided then to never put myself in a place again where anybody would take a tune away from me.' "
Years later, Scott found himself on an eight-week European tour playing with saxophonist Sanborn, performing the same song each night.
So despite Morgan's reputation as a hard-liner, his students frequently exhibit the highest reverence for him.
Walker credits Morgan with "changing my life." The bassist arrived at his HSPVA audition unable to read music, but Morgan saw something in the young instrumentalist. "Doc didn't give up on me," he says. "He gave me my career."
A full list of students who have gone on to do great work would take more space than is available. Though Walker makes his living playing with Jarreau, he had a Top 40 hit in 1992 with the single "Take Time." Moran earned a MacArthur Fellowship "genius grant." Glasper won a Grammy for his own work and can also be heard on "To Pimp a Butterfly," an almost unanimous best album of 2015 by rapper Kendrick Lamar. Drummer Chris Dave played in Houston earlier this year with R&B star D'Angelo, who released one of the best-reviewed albums of the past year. Drummer Reggie Quinerly put out a strong second album earlier this year, and guitarist Mike Moreno just released "Lotus," certain to be a well-received recording at the top of the new year.
Walker last year began organizing DocFest, an annual concert and fundraiser in which a cross-generational group of HSPVA jazz alums convene to show appreciation for their mentor while also putting on a concert headlined by a notable HSPVA grad followed by a spirited jam session.
In addition to being a top-shelf night of live music, DocFest raises money for the Helen and Bob Morgan Jazz Scholarship Fund at New York's New School for Jazz and Contemporary Music. Many of Morgan's students continued their education at the New School before going on to successful careers in music: Glasper, Quinerly, Jamire Williams, Alan Hampton among them. Though Morgan has stepped away from playing the part of educator, the scholarship keeps him involved in the development of music careers.
This year's headliner, drummer Herman Matthews, graduated in 1979 and left Houston for Los Angeles, where he has put together a remarkable résumé playing with R&B and jazz performers such as Stevie Wonder and the Isley Brothers, as well as a long run with the funk ensemble Tower of Power.
He'll be joined by trumpeter Nelson Morales, trombonist Rob Negrin, saxophonist Warren Sneed and keyboardist Mike Rojas - players who have performed and recorded with acts like Frank Sinatra, Celia Cruz, Freddie Hubbard, Doc Cheatum and many others.
Walker - whose work has ranged from R&B with Regina Bell to avant garde jazz with Ornette Coleman - will play bass.
"One person can change things in this world," Walker says. "That's what Doc did for us."


Nissan LEAF owners very satisfied
We have just a short article to post today, but one that we feel is important, especially given the nature of much of the dialog surrounding the Nissan LEAF over the past several months.
One point that we wish to highlight from the Chelsea Sexton/Andy Palmer YouTube video (found here) is that the Nissan LEAF has achieved the highest satisfaction ratings of all 64 Nissan vehicles sold globally. According to Mr. Palmer (right around the 5:40 mark in the video), LEAF has at or above a 95 percent satisfaction rating. When you think about it, that is an astounding satisfaction rating for any product.
A saying that we learned a long time ago in the automotive business is that nobody builds the perfect car. Compromises always exist as individual needs vary from household to household. This is exactly why such a broad array of vehicles can be found at any manufacturer’s retail dealerships – one driver’s needs are often dramatically different than anothers.
Our view is that the LEAF’s high satisfaction ratings bode well for the electric vehicle (EV) movement in general. The LEAF is certainly not a perfect car. But for those that present the right set of circumstances to acquire one, it seems to be a very good solution to their transportation needs, as self reported by owners satisfaction ratings measured by Nissan. As battery technology improves, prices fall due to manufacturing efficiencies, and driving range increases, we see a bright future for electric mobility as presented by Nissan and other automobile makers as they continue to broaden the scope of fully electric vehicle choices.


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The White House is promoting the extension of the Bush Tax Cuts for the middle class (which should really be called the Obama-Bush Tax cuts, since Obama extended them in 2010) as exactly that, tax cuts. But the truth is, an extension of the tax cuts wouldn't be a cut at all, but would keep tax rates at the current rate. The Obama admnistration is implying on the official WhiteHouse.gov website that a tax cut is coming for the middle class when in reality if a deal is made by January 1, middle class families won't see a tax...
In response to:
Cause we're too busy laughing at you getting :&)#% by George Soros!
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ATLANTA, Ga. (CBS46) – There is a heavy police presence Wednesday afternoon at Booker T. Washington High School in southwest Atlanta.
Several Atlanta Police officers are currently on scene and have cordoned off a portion of the campus.
Atlanta Public Schools issued the following statement regarding the incident:
Following a physical altercation between students during dismissal at Washington High School, a parent came to the campus with a weapon and began waving it in the direction of students, employees, and an Atlanta Public Schools Police Officer. The officer ordered the parent to drop the weapon. When the parent did not comply with the order, the officer fired one shot which struck the parent in the hand.
Paramedics were called to the scene to treat the parent’s wound. No students or school employees were harmed during the incident. APS Police and Atlanta Police are conducting a full investigation of the incident, and the parent faces several charges. As this was an officer-involved shooting, the Georgia Bureau of Investigation is investigating as well.
The safety and well-being of our students and employees is always a top priority for all Atlanta Public Schools.
Copyright 2022 WGCL. All rights reserved.


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All too typical pattern.
After being a socially active student, a former university friend notes Collin “moved back home (to Calgary) and started to be hardcore Muslim.”
That “hardcore” ideology has characterized Collin’s social media presence, as just two months ago he tweeted “Victory or Shahada (martyrdom) in the end win win.”
Canadian Brothers Latest in Islamic State’s ‘Foreign Legion’
By Dalit Halevy, Ari Yashar
Further testimony has been brought regarding the powerful recruitment pull of the Islamic State (IS, formerly ISIS) on the West, as two young Muslim converts from Canada have been identified in the ranks of the extremist terrorist group.
The brothers Gregory and Collin Gordon of Calgary recently converted to Islam, taking the names Abdul Malik and Khalid. Between 2011 and 2012, they lived in the same apartment that Damian Clairmont and Salman Ashrafi once lived in; the two both died fighting for IS.
Sources in the Calgary Muslim community told the Canadian CBC News that the two brothers disappeared in late 2012 – the same time that Calirmont and Ashrafi left for Syria – and since then the two have posted photos of themselves in military fatigues.
The radicalization of the brothers has left many who knew them perplexed; Collin was a student in the Southern Alberta Institute of Technology (SAIT) and Thompson Rivers University (TRU) where he studied business.


Yesterday the fine arts school in our community had an open house for kids where they could test choir, dance, instruments, and more so obviously I took Jambo.
I had high hopes for the choir session, but he wasn't that into it. Dance, however, was a hit, as were the instruments. He got to test a trumpet, trombone, piano, xylophone, and a drum kit. The latter was his favorite and he could keep a decent beat. Don't tell him (or my neighbors) but I put a mini drum set on his birthday list.


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Tree Planting Mission continues in southern Belize
YICE’s Nursery, in partnership with Barefoot Children Ministries, provided over 650 tree seedlings to remote, southern communities in 2019.
Ya’axché Institute for Conservation Education (YICE), the sustainable financing program of Ya’axché Conservation Trust, partnered with Barefoot Children Ministries on a tree planting mission to promote healthy and sustainable livelihoods in communities in southern Belize. During this planting mission, 388 fruit and timber seedlings, all sourced from our Nursery facility, were distributed to 180 families in 3 rural communities.
YICE’s Nursery was established to provide coffee, cacao and Inga seedlings to farmers in order to facilitate the agroforestry plantations. Since then, our nursery has been expanded to 2,928 square feet with the capacity to grow 55,000 seedlings. Today, our nursery not only supports farmers and other community members but also sustainable projects across Belize. One of those projects is that of Barefoot Children Ministries, a nonprofit corporation that is dedicated to improving the environment and nutrition of Belizean families in the Toledo district while facilitating their economic development.
YICE Busines Manager Marcelia Assi along with members of the Barefoot Children Ministries handing out seedlings. | Photo by: Marvin Vasquez
On June 6, 2019, the villagers of Conejo, Sunday Wood and Otoxha received a variety of seedlings including cedar, mahogany, jamaican lime, bilimbi and kinep. The distribution of timber and fruit species will support climate resiliency in these communities by enhancing carbon sequestration and sustaining livelihoods and food sovereignty. Upon receiving the seedlings, a short presentation was provided to children and their families on how to plant and take care of the seedlings so that the young trees can thrive. This was followed by a hands-on demonstration on how to transplant the seedlings from their bags. Our dedicated staff was more than happy to be part of this project; they especially enjoyed watching the excited children receive their seedlings.
Newly germinated cacao seedlings at Ya’axché’s Nursery. | Photo: Maximiliano Caal
This year, we have delivered 658 healthy organic trees to 305 families in 6 communities. YICE hopes to continue working with Barefoot Children Ministries and looks forward to the triannual distribution of seedlings to communities in the Toledo district. We welcome partnerships with individuals and organizations on projects that aim to bring sustainable livelihoods in Belize.
Our nursery is located at Mile 68 on the Southern Highway, Golden Stream Corridor Preserve. Contact us at 722-0108 or email [email protected] or visit us to purchase your seedlings or make your order.

domain: agronomy


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After giving his 39-year-old body some time to rest during the summer Steve Nash has been walking a fine line all preseason of trying to round into shape without pushing too hard and risk falling back into the cruel injury cycle that sabotaged his first season with the Los Angeles Lakers. With the regular season set to tip off in less than a week some of Nash's teammates are starting to wonder when or even if they'll have the full services of the surefire Hall of Famer in their lineup this season. Coach Mike D'Antoni brought it up first after the team's exhibition game against the Sacramento Kings in Las Vegas suggesting that Nash would be in and out of the lineup all season. Pau Gasol followed this week saying he was "concerned" about Nash after the point guard played only 15 minutes in the Lakers' win over the Utah Jazz sitting out the second half because of a neck injury. "Steve hasn't been able to complete a practice yet so that's the bigger issue" Gasol said of Nash who is averaging just 3.5 points and 3.5 assists in 16.7 minutes per game in the preseason sitting out one of the exhibition games altogether. "I'm a little bit concerned because I want him to be healthy I want him to play. I want him to play and I want him to do well. I want him to help us. I hope that he can." D'Antoni said Nash was able to make it through a full practice Thursday but cautioned that he could hold Nash out of the preseason finale Friday in Anaheim. If Nash doesn't play Friday does he think he will be ready for the regular-season opener Tuesday against the Los Angeles Clippers? "I do I do" Nash said. "I played the last game even though I probably shouldn't have trying to get a little extra rhythm and some reps. By the end of the week I should feel better." Nash said that had the Utah or Sacramento games been in the regular season and meant something he could have stayed in and pushed through his ankle and neck discomfort that caused him to shut down his night at halftime in both instances.
Nash's health starting to cause concern
ESPN Los Angeles | Oct 25


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MIAMI — WHAT IS THE EXCHANGE? IS IT THE SAME AS THE HEALTH MARKETPLACE?
The federally run health exchange or marketplace is an online shopping site designed to take the confusion and anxiety out of buying private health insurance for individuals and families who buy their coverage directly. Exchanges are meant to have the feel of an online travel site, like Expedia, where shoppers can compare prices of different plans. The marketplaces will be available in every state and are part of the Affordable Care Act.
The Obama Administration was referring to the online site as an exchange, but now prefers the term 'marketplace.'
WHAT ARE THE DIFFERENT WAYS I CAN ACCESS THE MARKETPLACE?
—The government unveiled a redesigned website, healthcare.gov, last month, which includes a web chat feature on the site.
—You can call 1-800-318-2596 24-hours a day, seven days a week and discuss various health plans with an operator in English, Spanish and other languages. Operators will take into account things like whether you have a disability, are pregnant, have a pre-existing condition or are a veteran to help narrow down what plans might be a good fit for you.
—You can also apply through regular mail by filling out a 13-page application.
HOW MUCH WILL THE AVERAGE HEALTH INSURANCE PLAN COST ME?
It's hard to say because Florida lawmakers voted to let the federal government run their marketplace. The feds still haven't published the rates because they are still finalizing them with insurers, but they are expected to be released in September. All plans will be required to offer the same set of benefits under the bronze, silver, gold and platinum plans, but the costs, including premiums, deductibles and co-pays are still a secret.
WHY DO CALIFORNIA RESIDENTS KNOW THEIR RATES, BUT FLORIDA STILL DOESN'T? ISN'T IT ALL UNDER THE SAME FEDERAL LAW?
The marketplaces are all under the same Affordable Care Act. But California is running its own marketplace so officials there already published their rates. Florida is at the mercy of the federal government for that information because the feds are running Florida's marketplace.
WILL I QUALIFY FOR A SUBSIDY?
Generally, those making less than $48,000 a year who don't have the option of getting health insurance through their jobs will qualify for some type of voucher from the government to help pay their insurance premiums. The less you make, the more the government will pay.
Kaiser Family Foundation has an interactive calculator that estimates premiums based on age, income, gender and family size as well as the tax subsidies that may be available. http://healthreform.kff.org/subsidycalculator.aspx
WHEN WILL ALL THIS TAKE PLACE?
Enrollment begins Oct 1. Coverage starts Jan 1. Enrollment ends March 31.
WHAT IF I CHANGE MY MIND AND WANT OUT OR WANT TO SWITCH TO ANOTHER PLAN?
Once you choose a plan, you're locked in for a year for the most part. If you get insurance through the marketplace and then get a job and want to switch to your employer's plan, you may have to pay a small penalty.
WHAT HAPPENS IF I MAKE BELOW THE FEDERAL POVERTY LEVEL? CAN I STILL GET COVERAGE UNDER THE MARKETPLACE?
No. Under the law, Medicaid is the only coverage option for people below the poverty line — $11,490 for an individual or $23,550 for a family of four. People this poor cannot get subsidized private coverage in the new health insurance markets.
Eighteen states, including Florida, aren't expanding their Medicaid programs.
The poor will be exempt from penalties for being uninsured, but they also won't get help with their health care.


The Sony Xperia Z3 has just passed China’s TENAA certification process, revealing details about two versions of the handset and their hardware specifications.
Do the Oppo Find 7’s improvements justify spending the extra $100? The device has amazing specs, and we have taken it through the most demanding tests.
Qualcomm showcases the power-saving capabilities of the Snapdragon 801 in music playback via Hexagon DSP, allowing continuous playback on the OnePlus One for 60 hours.
A series of benchmark tests demonstrate that, despite the QHD display, the Samsung Galaxy S5 LTE-A offers up similar battery life and performance to the regular Galaxy S5.
AnandTech’s Anand Lal Shimpi ran a suite of benchmarks on a Mobile Development Platform tablet provided by Qualcomm to see exactly what kind of performance the Snapdragon 805 brings to the game. His conclusions? It’s mostly about the next-gen graphics.
The launch of the OnePlus One is only two weeks away and even at this late stage the company is still fine tuning the device so that it will offer some of the best performance on the market. Peter Lau has announced that a 2.3 GHz Snapdragon 800 is too old school and instead the handset will use a 2.5GHz Qualcomm Snapdragon 801.
As the new Galaxy S5 and Xperia Z2 both come equipped with a new Qualcomm Snapdragon 801 SoC, let’s examine what this new chip offers over the old Snapdragon 800.


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Three years ago my husband and I had the good fortune of meeting a lovely British couple at the Ramstein Welfare Bazaar.
Bob Wallis is a retired policeman and for almost 10 years he was the commander of the Sub Division named Newmarket which included the civilian responsibility for the bases of RAF Mildenhall and Lakenheath. He retained the distinction of commanding the Mildenhall Air Show for 10 years which, at that time was the largest two-day air show in Europe.
His wife Janet is a retired school teacher who suggested they pursue their hobby “in a small way” which has since blossomed into an enjoyable and flourishing business.
At a recent bazaar, we wandered into Bob and Janet’s little booth and were given a friendly lesson in cigarette card history and have since become cartophilists (cigarette card collectors) ourselves.
Cigarette cards were given as free gifts inside cigarette packs and were uniquely introduced in the United States more than a century ago. Following complaints of damaged cigarettes in packs, American tobacco manufacturers came up with the idea of providing beautiful pictures on thick cards to be used as stiffeners inside the packs. The first sets included photographic cards of girl cyclists, actors, actresses, beauties and celebrated American Indian chiefs. The world’s first baseball cards were in fact American cigarette cards.
This unique tradition continued until 1903 when the beginnings of the Imperial Tobacco Company of Great Britain and Northern Ireland emerged. Thereafter cigarette cards were distributed worldwide throughout the former British Empire. With a few exceptions, cigarette cards finished in the United States in 1910 and in the United Kingdom (with similar exceptions), in 1940.
Cigarette cards reflect almost every subject imaginable. The dates printed on the back are well researched and the cards are high quality. This information poignantly and accurately describes history and they are, in themselves, educational. Some of the most popular cigarette cards include illustrations of architecture, flowers, military subjects and sports. Cigarette cards convey truth, beauty and culture by picture and the printed word.
Cigarette cards are truly a historic part of the tobacco manufacturing industry and their value will substantially increase over the next decade or so.
Bob and Janet have been regulars at the Ramstein Bazaar since 1997. They said the largest collectors of cigarette cards are often school teachers. What Bob and Janet have found most amusing is that Air Force families often purchase Army and Navy cards and Army folks collect aeroplane sets. No particular category is more collectible than others. New collectors are always welcome, and Bob and Janet will cheerfully explain the nature and range of this hobby.
For more information, e-mail bob-gaze
firstname.lastname@example.org or check out their
Web site www.gazeleyccs.com.


Political talk in Alwar veers towards cows
“It’s not the reality of Rajasthan, it’s the reality of the world,” Rajasthan chief minister Vasundhara Raje said recently.
Man beaten to death in Alwar on suspicion of smuggling cows
Akbar Khan, 28, and another man were taking two cows to their village in Haryana ...
SC asks Centre, states to comply with directives on cow vigilantism, mob lynching
The Supreme Court Monday asked all states and Union Territories to comply with its directions on curbing cow vigilantism and mob lynching, a...
Government panel to study role of cow urine, dung in curing diseases
The central body - a national steering committee for scientific validation - will evaluate whether 'Panchagavya' - a concoction of cow urine mixed with cow dung, milk.
What made rural India abandon its cattle in droves
An extraordinary situation is prevailing in rural India. Stories of cattle distress are aplenty.
Cow protection needed to bring change in country: Minister at launch of 'Muslim Gau Raksha Sangh'
The social justice and empowerment minister termed formation of the outfit as a "revolutionary step" that will bring Hindus and Muslims closer.
Cow dung neutralizes radioactive elements: Rajasthan minister Vasudev Devnani
After a session on spiritual and scientific importance of cows for humans, the minister moved on to explain its importance on farming.
Onus on states to check cow vigilantism: Supreme Court
A bench of Chief Justice Dipak Misra AND Justices A M Khanwilkar and D Y Chandrachud said this was a law and order issue and each state has to be responsible.
Villagers question 4 names in FIR on cow slaughter, 11 & 12 yr old among those named
Other villagers also disputed the names of two men in the FIR, saying one did not live in the village and the other was working in the Ijtima at Bulandshahr
No cash cow, Army to sell bovines for token amount
While the special Frieswal cows are believed to be valued at over Rs 1 lakh each, the lack of buyers has finally forced Defence ministry's hands.
Tension grips Bulandshahr after police officer, civilian killed over alleged cow slaughter
The Uttar Pradesh Police on Tuesday arrested two men and detained four in connection with a mob violence in Bulandshahr district that led to...
ET View: Bring about an anti-mob law, and enforce it with seriousness
Vigilantism is the result of perceived and real neglect of the State not conducting its duty of keeping the law, and thereby allowing this task to be ‘outsourced’.
Telangana Polls: BJP releases party manifesto, promises cows to 1 lakh people every year
Farm loan waiver up to Rs two lakh, free laptops to degree students, regulation of liquor sale, free distribution of cows to one lakh people...
Copyright © 2019 Bennett, Coleman & Co. Ltd. All rights reserved. For reprint rights: Times Syndication Service