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The binding of Steel factor (SF) to c-kit initiates a signaling pathway essential for development and maintenance of interstitial cells of Cajal (ICC). Soluble and membrane-bound isoforms of SF are expressed in the gastrointestinal tract, but the role for either isoform in supporting ICC development is unknown. The aim of this study was to determine the role of SF in supporting ICC in culture. ICC were cultured from dissociated mouse jejunum and grown with fibroblast cell lines that produced either soluble, membrane-bound or membrane-restricted SF. ICC were identified and counted by c-kit immunoreactivity. The number of c-kit immunoreactive cells was greater in the coculture system compared with cultures grown without SF-producing fibroblasts. All forms of SF-producing fibroblasts increased ICC number in culture but physical separation of the fibroblasts from the c-kit immunoreactive cells, the addition of exogenous SF to the culture medium, or fibroblast-conditioned media did not. These results are consistent with the hypothesis that the membrane-bound form of SF preferentially contributes to expression of c-kit-positive ICC under cell culture conditions.
- immunocytochemistry
- gastrointestinal pacemaker
- Steel-factor proteolysis
the binding of steel factor (SF) to the receptor tyrosine kinase c-kit initiates a signaling pathway in interstitial cells of Cajal (ICC), which is essential for the normal development of ICC and rhythmic activity in the gastrointestinal tract (14, 17, 20, 21,37). ICC function as pacemaker cells in the gastrointestinal tract and may also modulate enteric neurotransmission (15, 28,29). Spontaneous mutations at the white spotting locus, which encodes the c-kit receptor or the Steel locus, which encodes SF, are associated with disruption of the ICC network located at the myenteric plexus region (ICC-MY) of the small intestine and the functional loss of the spontaneous electrical slow wave and contractile activity (23, 24, 34, 35). SF activation of c-kit is also necessary to maintain ICC, because the administration of neutralizing c-kit antibody results in the subsequent disappearance of ICC (21,30). At the mRNA level, SF and c-kit expression are temporally correlated with SF expression preceding c-kit expression and with both peaking between embryonic days 13 and 15 in the murine small intestine (37). Thus SF/ c-kit signaling is required for both ICC development and for normal gastrointestinal function.
The KITLG gene encodes two distinct isoforms of SF, and both are synthesized as transmembrane proteins expressed at the cell surface (12). Proteolytic processing produces biologically active soluble SF, but the rate of cleavage of SF from the two isoforms differs. One isoform, known as the soluble SF, is characterized by rapid proteolytic cleavage and release from the plasma membrane. The other isoform is known as the membrane-bound SF isoform, because it lacks the proteolytic cleavage site encoded by exon 6 in the KITLG gene and releases soluble SF much more slowly (27). Therefore, both isoforms initially produce membrane-bound SF and the main difference is the rate of release of soluble SF. The membrane-bound SF isoform contributes higher steady-state levels of membrane-bound SF, whereas the soluble SF isoform contributes more significantly to steady-state levels of soluble SF. Each isoform is thought to play a specific physiological role, because the ratio of the membrane-bound-to-soluble SF varies in different tissues (12). Separation of these isoforms occurs in the spontaneous Steel-Dickie (Sld) mutant mouse that exclusively expresses soluble SF, possibly providing an indication for the role of membrane-bound SF (2). The Sl/Sldmouse does not display a spontaneous, rhythmic electrical slow wave, and ICC are not present in the myenteric plexus (34). These observations suggest that membrane-bound SF provides an essential role for the development and/or maintenance of ICC-MY. Interestingly, ICC populations in the colon and the distal stomach appear normal in Sld mice, which indicates that membrane-bound SF is not required for the development of these ICC populations (34).
The physiologically relevant form of SF supporting ICC in humans is not clear. SF is present in low concentration in serum but is thought to act locally, close to the site of production where the concentration is likely much higher (2). Soluble SF circulates as a dimer and a monomer, but the dimeric form is more biologically active (11). Membrane-bound SF is a more effective agonist for the c-kit receptor compared with soluble SF (25). Stimulation of c-kit with membrane-bound SF in a SF-dependent myeloid cell line MO7e resulted in more persistent activation of c-kit kinase compared with stimulation with soluble SF (25). Therefore, it is possible that membrane-bound SF may activate c-kit on ICC more effectively than soluble SF.
Elucidating the role of membrane-bound vs. soluble SF in the SF/c-kit signaling pathway in ICC is important, because loss of ICC is observed and may be involved in the pathophysiology of several motility disorders, such as slow transit constipation (8), diabetic gastroenteropathy (9), and pseudoobstruction (38). Loss of ICC may be a primary event or secondary to loss of a signaling molecule, such as SF, from a specific cell type in the gut. Fibroblasts genetically engineered to produce soluble SF or only membrane-restricted SF have been used to differentiate the role of membrane-bound SF in hematopoietic tissue. With the use of these modified fibroblasts, Miyazawa et al. (25) demonstrated that, compared with soluble SF, membrane-bound SF induced more persistent activation of c-kit and increased the lifetime of activated c-kit complex at the plasma membrane. In addition, membrane-bound SF induced greater proliferation of an erythrocytic progenitor cell line compared with the soluble isoform of SF (16). These results suggest that soluble SF and membrane-bound SF play different physiological roles in hematopoiesis.
The aim of this study was to test the hypothesis that the membrane-bound form of SF is required for ICC development and survival. Primary cell cultures from the murine jejunum were used as a source of ICC and cocultured with murine fibroblasts that express soluble SF, membrane-bound SF, or membrane-restricted SF. The results suggest that local expression of SF is required for successful culture of ICC.
Primary cultures were derived from freshly dispersed cells from the murine jejunum. BALB/c mice (Harlan Sprague-Dawley) of either sex between 9 and 15 days old were killed with CO2 inhalation. The small intestine was removed and pinned out in a Sylgard-lined dish containing Hanks calcium-free buffer and 1% antibiotic-antimycotic (GIBCO). The muscularis propria was gently peeled from the mucosa and submucosa and placed in a collagenase-based dissociation cocktail. The cocktail contained 8 mg collagenase (model 4176; Worthington), 20 mg bovine serum albumin (model A-7511; Sigma), 20 mg trypsin inhibitor (model T-9128; Sigma), 5 mg adenosine triphosphate (model A2620–9; Aldrich) and 10 ml of calcium-containing Hank's balanced salt solution. The pH was adjusted to 7.0 with 0.1 M NaOH. After 15 min of incubation at 32°C in a gently shaken water bath, the tissue was washed twice with fresh calcium-free Hank's balanced salt solution and returned to 32°C. The tissue was then gently triturated every 3 min until single cells were obtained, for ∼10 min. Cells were washed and resuspended in 12 ml of smooth muscle basal medium (Clonetics). This preparation likely contains ICC-MY, ICC in the deep muscular plexus region (ICC-DMP), and ICC distributed throughout the muscle layers (ICC-IM). The majority of ICC in culture is thought to be ICC-MY, because this region contains the largest number of ICC. Also, the majority of c-kit-positive cells in culture exhibits branching processes, similar to ICC-MY in situ. However, the exact proportions of ICC-MY, ICC-DMP, and ICC in the smooth muscle in the cell dispersions or the primary cultures cannot be determined at present, because all cells studied were c-kit positive. It is currently unclear whether the local environment and presentation of SF favor a particular ICC class or influence morphology and physiology.
Murine jejunal cell culture and murine cells/fibroblast coculture.
Freshly dispersed cells obtained from the murine small intestine were cultured on 25-mm glass coverslips at a cell density of ∼5 × 104 cells/ml. Coverslips with established murine fibroblasts covering approximately one-third of the surface were used for cocultures with freshly dissociated murine jejunal cells. Control murine fibroblasts and fibroblasts genetically modified to express one of the SF isoforms were generously provided by David Williams (Indiana University School of Medicine, Indianapolis, IN). Four different fibroblast cell lines were used to provide soluble, membrane-bound, membrane-restricted, or no SF, respectively. A schematic of the different forms of SF and the potential role for each type is shown in Fig. 1. SF exists in several forms in vivo including soluble monomeric and dimeric forms, as well as a membrane-bound form (Fig. 1 A). Each form is capable of activating c-kit, but the efficacy and the specific effects are varied (25, 26). Membrane-bound SF persistently activates c-kit, compared with soluble SF, and the dimeric form of soluble SF preferentially stimulates mast cell growth compared with monomeric soluble SF (25, 26). In the present study, the role of membrane-bound SF on ICC cultures was assessed utilizing three types of fibroblasts genetically engineered to produce soluble SF, membrane-bound SF, or membrane-restricted SF (Fig. 1 B). A fibroblast line that does not express SF was used as a control. Soluble and membrane-bound SF have different proteolytic cleavage sites. However, both soluble SF-producing fibroblasts and membrane-bound SF-producing fibroblasts contribute to soluble SF levels. Membrane-restricted SF will not contribute to soluble SF, because it lacks the sequence where the proteolytic cleavage sites are located. Murine fibroblasts were immunostained with an antibody that recognizes SF to show that the fibroblasts do produce SF (Fig. 1 C).
Fibroblasts were plated on 25-mm plain glass coverslips (Fisherbrand) at 3 × 104 cells/ml in high-glucose Dulbecco's modified Eagle medium containing 10% fetal bovine serum, 1% sodium pyruvate, and 1% antibiotic-antimycotic (all from GIBCO). After 12 h, fibroblast cell division was arrested by treatment with mitomycin C (5 μg/ml) for 90 min to control the total number of fibroblasts per coverslip. After a 24-h recovery period, freshly dissociated jejunal murine cell suspensions (1.5 ml) were plated onto fibroblasts. The cell culture medium was changed after 24 h. Cultures appeared to be robust, i.e., spontaneous contractions of smooth muscle cells were observed. These contractions were maintained for at least 72 h. For these studies, all cell counts were performed 48 h after the murine cell suspension was plated. This time point was optimal, because ICC and c-kit expression appeared to be most stable. More time in culture allowed other undifferentiated fibroblast-like cells to overpopulate the cultures, and at shorter times, cell attachment was variable.
ICC were identified with the use of the rat monoclonal anti-c-kit antibody ACK2 (GIBCO) after cultures were briefly fixed in acetone (4°C for 10 min). Immunostaining using the antisera SCF G-19 (Santa Cruz Biotechnology, Santa Cruz, CA) or anti-mouse SCF (R&D Systems, Minneapolis, MN) verified SF expression. Acetone-fixed cultures were washed with phosphate-buffered saline (0.1 M; pH 7.4), incubated with blocking solution (10% normal donkey serum) for 1 h to minimize nonspecific antibody binding, and then incubated with primary antibody (5 μg/ml) for 24 h at 4°C. Cultures were then rinsed in phosphate-buffered saline and incubated for 2 h at 4°C with donkey anti-goat IgG, donkey anti-mouse Ig, or donkey anti-rat IgG conjugated to CY3 or fluorescein (1:200 dilution; Chemicon). Nonspecific immunoreactivity was assessed by immunostaining cultures in an identical manner except that the primary antibody was omitted. Immunostained cell cultures were examined with the use of a laser scanning confocal microscope (model LSM 510; Zeiss). A 40× (numerical aperature = 1.2) water immersion objective was used, with additional electronic zoom, when necessary. The full width at half-maximum signal intensity was ∼1.3 μm and, therefore, out-of-plane fluorescence was negligible. An excitation dichroic mirror was used with a bandpass emission filter of 530 ± 15 nm and a 590-nm long-pass filter. Images were reconstructed from confocal stacks of Z-series scans of 10–30 optical sections through a depth of 5–15 μm. The immunostained cultures were surveyed for c-kit-positive cells with the use of an algorithm to automatically move the stage in the X-Y plane in a 1 × 1-mm grid pattern. The number of c-kit-positive ICC per field was recorded at 50 intersections under direct observation. Number values reported in the text refer to the individual experiments. Each experiment was carried out with the use of tissue obtained from two mice; Three cover slips were used for each experiment and for each control. Fifty high power fields were counted for each coverslip. Occasionally, clumps of ICC containing more than six overlapping cells were found. However, no difference was noted with clump frequency in cultures grown with fibroblasts expressing soluble, membrane-bound, or membrane-restricted SF. The larger networks (>6 cells) were omitted from the total ICC cell count to be certain that increased counts of ICC did not result from undigested tissues. Parallel cultures, grown under identical cell culture conditions but without fibroblasts, were used to normalize ICC counts. A parallel culture was grown for each jejunal cell dispersion to allow normalization between cell cultures derived from different animals.
Data are expressed as means ± SE. Differences in data were evaluated by Student's t-test. P values <0.05 were taken as statistically significant.
Primary cultures yielded single cells and small clumps of cells. Spontaneous contractions of smooth muscle cells were observed betweendays 1 and 3 in small networks containing cells with ICC-like appearance (e.g., triangular cell bodies and multiple processes) and with smooth muscle-like appearance (7, 13,17). Positive immunoreactivity with the anti-c-kit antibody ACK2 confirmed that the cultures contained ICC. However, the density of ACK2-positive cells was low. The mean number of ACK2-positive cells (0.23 by 0.23 mm; expressed as number of ICC per high-powered field) under these control-culture conditions was 0.85 ± 0.2. One typical cell culture containing ICC is shown in Fig.2. Transmitted light images at low magnification (Fig. 2 A) show many cells with triangular cell bodies and several processes. Immunohistochemical staining revealed a much smaller subset of ACK2-positive cells, thus verifying ICC presence (Fig. 2 B).
Freshly dispersed murine jejunal cells grown with fibroblasts expressing membrane-bound SF contained more c-kit-positive cells compared with controls grown without fibroblasts producing membrane-bound SF. One typical cell culture is shown in Fig.3. Single ACK2-positive cells were distributed throughout the cultures (Fig. 3 A), and ICC networks were also common (Fig. 3 B). In the regions with ICC, the ICC density was much higher compared with control cultures grown without fibroblasts producing SF. Overall quantification of ICC showed an increase of 119 ± 14% (P < 0.05,n = 5) in coculture with fibroblasts expressing membrane-bound SF compared with control cultures grown without fibroblasts. ACK-2 immunoreactivity was not observed in pure fibroblast cultures (data not shown).
Both soluble and membrane-bound SF can stimulate c-kit in vitro, but soluble SF does not support long-term hematopoietic stem cell growth (11). To determine whether soluble SF was sufficient to support ICC, freshly dispersed murine jejunal cells were cocultured with fibroblasts expressing soluble SF. Appearance of these cultures was similar to those grown with membrane-bound SF with many single ICC and ICC networks identified by ACK2 immunoreactivity (Fig.4). Freshly dispersed murine jejunal cells grown with soluble SF-secreting fibroblasts yielded an 80 ± 16% increase in ICC (Fig. 4 B) compared with control cultures (P < 0.05, n = 6).
Both isoforms of SF initially produce membrane-bound SF, and, to different degrees, both produce soluble SF (11). To distinguish the effects of soluble from membrane-bound SF on ICC in culture, we used a genetically engineered form of SF known as membrane restricted. This form of SF lacks the amino acid sequence to allow proteolytic processing and, therefore, does not release soluble SF. Freshly dispersed murine jejunal cells grown with fibroblasts expressing membrane-restricted SF are shown in Fig.5. The appearance of these cultures was qualitatively similar to those grown with either soluble or membrane-bound SF. The effect of membrane-restricted SF on the number of cells expressing the c-kit receptor is shown in Fig. 5 B. Freshly dispersed murine jejunal cells cocultured with fibroblasts expressing membrane-restricted SF resulted in a 180 ± 43% increase in c-kit-positive ICC compared with control cultures (P < 0.05, n = 3). To determine whether one isoform of SF more effectively increases the number of c-kit-positive ICC, membrane-restricted or soluble presentation of SF was tested in parallel cultures. Membrane-restricted SF did not increase c-kit-positive ICC expression more than that observed with fibroblasts expressing soluble SF (−9 ± 14%, P> 0.05, n = 3; Fig. 5 C).
Soluble SF is presented in membrane-bound form before proteolytic cleavage and, therefore, it was possible that the stimulation of ICC by soluble SF-producing fibroblasts results from membrane-bound SF. To test this hypothesis, fibroblasts were grown on permeable supports to physically separate the freshly dispersed murine jejunal cells from fibroblasts expressing soluble SF. Separation of the cocultures with the Transwell barrier prevented stimulation of ICC (Fig.6 A). The number of c-kit-positive ICC grown in cocultures separated by a semipermeable barrier (pore size = 0.45 μm) was unchanged from control cultures (−16 ± 11%, P > 0.05,n = 3) grown without fibroblasts. To test whether an increased concentration of SF could enhance the number of c-kit-positive ICC recombinant soluble murine, SF was added to the culture medium. The addition of 20 or 100 ng/ml did not stimulate expression of c-kit-positive ICC compared with control cultures (37 ± 19% and 17 ± 14%, respectively, P> 0.05, n = 3; Fig. 6 B). This suggests that enhancement of c-kit-positive ICC required close contact between the fibroblasts and the freshly dispersed murine jejunal cells.
Fibroblasts are support cells that enhance cell proliferation and survival by several mechanisms, and it is possible that stimulation of c-kit-positive ICC cell counts in these cultures could be independent of SF. This was tested with the use of fibroblasts that do not produce SF for coculture with freshly dispersed murine jejunal cells. Under these conditions, expression density of c-kit-positive was very low (Fig. 7 A) and the total number of c-kit-positive ICC observed under coculture conditions was not different from control cultures grown without fibroblasts (−11 ± 11%, P > 0.05, n = 3; Fig.7 B). The specificity of SF-stimulation through the c-kit pathway was also tested with the use of ACK2 to block c-kit activation. The addition of ACK2 to the culture medium of freshly dispersed murine jejunal cells with fibroblasts expressing soluble SF resulted in decreased expression of c-kit-positive ICC (Fig.8 A). The number of c-kit-positive ICC was reduced in a dose-dependent manner with a 54 ± 10% decrease after the addition of 5 ng/ml ACK2 and 74 ± 6% (P < 0.05, n = 4) decrease after the addition of 10 ng/ml (Fig. 8 B).
We have shown an increase in the number of c-kit-positive ICC when mouse jejunal cells were cocultured with SF-producing fibroblasts. Increased c-kit-positive ICC specifically resulted from fibroblast SF expression, because few c-kit-positive ICC were found in cocultures with fibroblasts that did not express SF. The c-kit-specific antibody ACK2, which interferes with SF activation of the c-kit receptor, also reduced the number of c-kit-positive ICC in culture, further suggesting a specific SF-c-kit interaction needed for survival of ICC. Effective culture of ICC required local production of SF, because physical separation of the jejunal cells from the fibroblasts or the addition of recombinant SF to the cell culture media reduced the relative number of c-kit-positive ICC in culture. In addition, conditioned media from fibroblasts synthesizing soluble SF did not support ICC in culture (data not shown). These results showed that the location of SF production is critical, with close contact between the c-kit receptor expressed on ICC and SF expressed on fibroblasts necessary for ICC expression. Data also suggest that in the gastrointestinal tract, required SF needs to be provided by membrane-associated SF on closely apposed supporting cells rather than by SF secreted from more distant cells or in the blood plasma. One alternative explanation is that presentation of membrane-bound SF merely enhances expression of c-kit by ICC but doesn't increase the number of ICC. For example, ICC with c-kit expression below detectable limits would not be identified under control conditions and coculture with SF-expressing fibroblasts would enhance c-kit expression in those cells to detectable levels.
Stromal cells, fibroblasts, and endothelial cells express SF as a membrane-bound protein (2, 10, 19). Soluble SF results from the proteolytic cleavage of an extracellular portion of SF (5, 22). Binding of SF to the tyrosine kinase receptor c-kit results in rapid receptor autophosphorylation, endocytosis, and degradation, thus regulating surface c-kit density and the duration of c-kit signaling (1). Although soluble and membrane-bound SF activate c-kit, membrane-bound SF stabilizes the receptor at the cell surface slowing receptor internalization and prolonging c-kit signaling (18, 25). Thus the membrane-bound isoform of SF increases the life span and the downstream signaling activity of the c-kit receptor. In contrast, soluble SF induces rapid downregulation of cell surface c-kit expression and a correspondingly rapid window of kinase activity (25). Recent studies have shown that the duration of c-kit activation may also function to differentiate specific downstream signaling elements (16). These data suggest that membrane-bound SF may persistently activate c-kit expressed on the ICC cell surface and stimulate specific downstream signaling pathways that ultimately enhance ICC differentiation and proliferation.
Development of ICC-MY in the murine small intestine depends on functional SF/c-kit signaling, appearing at embryonic day 13(17, 31, 34, 37). However, ICC expression may not be solely dependent on this signaling pathway, because mutant mice with impaired c-kit or SF signaling exhibit ICC-DMP and ICC in the colon and stomach (14, 35). This suggests that ICC-MY may be more dependent on SF signaling compared with ICC-IM or ICC-DMP. In this study, the number of ICC was much higher in cocultures with fibroblasts presenting SF and was thought to be mainly ICC-MY on the basis of percent and morphological observations. Complete disruption of the SF/c-kit signaling pathway results in the abolishment of ICC suggesting that the spontaneous mutant mice have developed compensatory mutations enabling ICC development by another pathway, or that a small residual c-kit activity is sufficient for certain classes of ICC (30).
The source for SF, as well as the relevant isoforms necessary for the development and maintenance of ICC in mature mice, is currently unknown. Very low numbers of ICC were observed in these studies in cultures without the addition of recombinant SF or fibroblast cocultures that supply SF. Survival of c-kit-positive ICC in these cultures may depend on SF production by another cell type that survives the enzymatic dissociation procedure. This could explain why increasing plating density in primary cultures improves ICC expression (37). One potential source for SF production is enteric neurons. These cells are ideal candidates for providing SF and stimulating ICC development and survival, because enteric neurons form close anatomical relationships with many ICC (33). Enteric neurons have been shown to produce mRNA for SF (32), but it is not known whether SF is expressed on the neuronal processes that intercalate with ICC. Murine knockout models suggest that enteric neurons are not required for ICC development. For example, the glial cell-line-derived neurotrophic factor (GDNF) knockout mice lack the enteric nervous system but exhibit normal ICC networks and mRNA expression levels of SF similar to wild-type mice (36). ICC are also normal in the c-ret knockout mouse, which lacks the GDNF receptor and the enteric nervous system (37). Another potential source of SF is smooth muscle. Recent data (4) showed that smooth muscle cells from the murine small intestine express mRNA for the soluble isoform of SF, and smooth muscle cells isolated from the gastric fundus express the membrane-bound isoform of SF mRNA (4). The authors (4) also found that single ICC, selected from the murine gastric fundus or small intestine, express the soluble isoform of SF mRNA. Thus it is possible that smooth muscle provides SF necessary for ICC development and maintenance. Because ICC develop from the same precursor cell as smooth muscle, ICC may produce SF that acts in an autocrine manner for the SF/c-kit signaling pathway. Such a mechanism has been reported for mast cells (3) and for neural crest cell development (6). Finally, it is well known that fibroblasts and endothelial cells express SF to support hematopoiesis (1,2), and therefore it is also possible that fibroblasts within the tunica muscularis express SF and support ICC.
Proximity between the cell type producing SF and ICC may affect both ICC density and morphology. ICC development in cell culture of dispersed murine intestine improves as plating density is increased (37), suggesting that the enzymatically dispersed cells are the likely source of SF. The authors (37) hypothesize that membrane-bound SF expressed in the myenteric plexus region by enteric neurons or fibroblasts could promote ICC proliferation and survival in this region, resulting in the dense network of ICC vital for spontaneous electrical activity in the gastrointestinal tract. This hypothesis also predicts the expression of bipolar ICC with smaller total volume and the lack of ICC network formation within the longitudinal muscle layer where a nonneuronal source of SF supports ICC development (37). Smooth muscle has been shown to preferentially express soluble SF, which is consistent with development of intramuscular ICC due to a brief stimulation of c-kit (4).
In summary, the present results show that coculture of freshly dispersed jejunal cells from the mouse with SF-expressing fibroblasts enhances the expression of ICC. Increase in ICC number in these cocultures is consistent with enhanced activation of c-kit, resulting from an increase in the local concentration of SF. Results suggest that membrane-bound SF potently stimulates the c-kit receptor and maintains the ICC phenotype. Soluble SF may promote very transient stimulation of c-kit, resulting in rapid receptor turnover and subsequent downregulation, whereas membrane-bound SF may persistently stimulate c-kit, promoting more robust ICC growth, proliferation, and survival. Identification of the specific expression pattern during development and later in life for each isoform of SF, as well as the cell type that expresses SF and supports ICC development, proliferation, and growth, will further determine the functional role for each SF isoform.
The authors thank Jan Applequist for support in preparing the manuscript and Jim Tarara for assistance with confocal microscopy.
This study was supported by National Institute of Diabetes and Digestive and Kidney Diseases Grants DK-57061, DK-52766, and DK-17238.
Address for reprint requests and other correspondence: A. Rich, Bristol-Myers Squibb, Bldg. 21, Rm. 1318, 311 Rockyhill-Pennington Road, Pennington, NJ 08534 (E-mail:adam.rich{at}bms.com).
The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.
First published October 9, 2002;10.1152/ajpgi.00093.2002
- Copyright © 2003 the American Physiological Society


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Mongoose Men's Status 2.4 27.5″ Wheel Full Suspension Bicycle, Steel Blue, 18″/Medium Frame Size
The Mongoose status 2.4 27.5 inch men's full suspension mountain bike features a light weight aluminum frame and alloy rims plus 21 speeds for all types of riding. Linear pull rear brake and disc front brake provide great braking power and it's designed with a comfortable but athletic riding position. Product Features Aluminum full suspension
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text: Well all, these are my two choices. I won't be buying for another two weeks, but wanted your opinions. Obviously the major pros for each are that the 45A supports DVD-A and SACD, but the Phillips supports SACD and upsamples redbook cds to 96/24 as I understand it. Thus my question is this: Is the quality of playback on redbook cds really that much better on the phillips than the Pioneer? Will it be a noticable improvement over playback on the 45A? Will I be kicking myself down the road later if I decided to not go with a universal player (no DVD-a)? In a nutshell, will the quality of redbook cds be improved enough to merit the loss of the DVD-A format? Assume they are the same price. As I understand it, the Elite 45tx reciever that I have ordered only has 1 multichannel input, so the addition of a stand-alone DVD-A player in the future is out. Please shed some light. THanks


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April 11, 2012 in Software (E)
[prMac.com] Vancouver, British Columbia - Outlisten, an innovative new platform for crowd-sourced concert videos that Wired Magazine called 'a new music format,' has chosen CloudEyes - a media synchronization technology that lets producers and contributors share and mine footage remotely - as the foundation for their offering.
"At concerts, everyone has a camera in-hand. We asked ourselves - what's the best way to share all that footage?" said Jeff Ponchick, CEO. "Uploading disconnected pieces of video to the web isn't good enough. We wanted multi-camera live concert videos that were fan-made, fan-sourced, and fan-edited, and with an interactive viewing experience. We were thrilled to discover CloudEyes, a fantastic technology that makes it not only possible, but easy."
The automatic synchronization technology of CloudEyes is used by video editors thousands of times every day to create everything from wedding videos to commercials to indie films. Like Singular Software's family of post-production tools, it syncs without the need for timecode, clappers or any special preparation, and automates the most time-consuming of media preparation tasks - except now, all those production benefits have moved from desktop to cloud.
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Visitors to Palm Springs love to party, so it's more than appropriate that each Spring the city is host to the Club Skirts Dinah Shore Weekend (aka The Dinah)—the largest lesbian event in the world.
This year marks The Dinah's 27th annual celebration and it kicks off at the close of Women's History Month on March 29 and continues through April 2.
"I was not the first lesbian to throw a Palm Springs Dinah party, but I was the first to see how big this event could get and started to build the international vision," says Mariah Hanson, the founder and producer of The Dinah. "It's the perfect spot to host the largest lesbian event in the world. I just knew if I built it, they would come. And they did—in droves! We're very proud to host the event in such a LGBTQ friendly city—a city that simply embraces diversity and recognizes that our differences are our strengths."
Hanson has always had a personal commitment to helping the LGBTQ community obtain equal rights and was on the board of Equality California during a pivotal time when she was one of the board members who pushed the organization to fight for the right to marry now, rather than wait until the overall community felt the timing might be more beneficial "We as a board decided that the time for civil rights is always now and that we must never wait for our equality, for our freedoms, and for our right to a life well-lived, free of fear and discrimination," she says.
This commitment to the community resonates with today's Dinah goers who Hanson says now live in a more tolerant world that embraces their lifestyle choices and celebrates who they are. And she notes that's reflected in the way the weekend unfolds: "The collective self-esteem of our community is very high and The Dinah celebrates that notion and helps to further increase our worth through an open and very loud celebration of our uniqueness."
And this year's event will indeed be a very loud celebration of that uniqueness starting with music from Keala Kennelly, the world surf champion turned cutting-edge DJ. "Lizzo is our choice for the next big thing as well as a group called FHB," adds Hanson. "They're very mysterious and that is creating some cache amongst the influencers. No one even knows what they look like."
Other performers include the rapper Tish Hyman, Latin hip hop artist Lady Cultura, stand-up comedian Julie Goldman, British comic Gina Yashere, and the 90s icons CeCe Peniston and Black Box.
"It's a well-rounded weekend packed with great talent and thousands of women," says Hanson. "Palm Springs—are you ready?!"
For more information, go to: www.thedinah.com
Photograph of Mariah Hanson: Courtesy Club Skirts Dinah Shore Weekend
This article rocks Mariah!! Sending highest energy for the most amazing Dinah ever!
With love,
Patty K.
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text: Phoenix Coyotes Renamed Arizona Coyotes 7:49:00 AM Arizona , Headlines , News Edit FFICE: It would appear that the name change will happen pretty quick though they are still officially the Phoenix Coyotes at this moment. The logos on the ice suggest that there will be a change sometime this season as they have removed the words "Phoenix Coyotes" from center ice. Share on Facebook Share on Twitter Share on Google Plus RELATED POSTS Phoenix Coyotes Renamed Arizona Coyotes Reviewed by DTC on 7:49:00 AM Rating: 5


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The Playful Dino wooden toddler activity toy is packed full of exciting activities! Multiple play surfaces, bead runs, peg mazes, and spinning dials create hours and hours of exploration and discovery, while encouraging cause and effect learning and fostering fine motor skill development. For over 30 years, Manhattan Toy has been making award-winning, high quality, educational toys for your baby, toddler or kid. From preschool activity centers and learning toys, to classic wooden toys for toddlers, our goal is to provide the safest and best toys available. All of our products, from the newest concepts to our time-tested classics, are innovatively designed to inspire imaginative play and delight our customers.
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DOVER, Del. — A Dover councilman has been censured by the City Council for violating "the spirt and purpose" of the city's ethics code.
The City Council voted 6-3 Monday to censure Councilman Brian Lewis.
Lewis said he was not given a chance to defend himself. He told the Delaware State News that he believes his rights were violated.
The formal reprimand came in response to Lewis' public questioning of whether fellow Councilman Tanner Polce should have participated in a council vote involving his part-time employer, Wesley College. The city solicitor had already issued his opinion that the councilman did not violate any ethical rules were he participated in the vote.


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Comments:02.02.2016 at 03:40 Mejora:
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Sql server import export access
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Comments:27.01.2016 at 21:44 Meztigul:
It cannot be!


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Cannot Call Methods On Nvarchar Asp.net
You cannot rate topics. This method or property cannot be called on Null values3Cannot call methods on nvarchar0Getting MySQL Trigger Error-1getting values from stored procedure0I am executing following procedure in oracle but get compilation error0How You cannot delete your own posts. Add a Solution Add your solution here B I U S small BIG code Plain TextC++CSSC#Delphi / PascalF#HTML / XML / ASPJavaJavascriptObjective-CSQLSwiftPerlPHPPythonVBXMLvar < > & link [^] encode untab case indent http://mobyleapps.com/cannot-call/cannot-call-methods-on-nvarchar-max-sql.html
Are 14 and 21 the only "interesting" numbers? You cannot edit HTML code. Monday, November 7, 2011 cannot call method on nvarchar - In my case, the issue was because i had typed a 'dot' instead of 'comma' - However, this may happen due Consequences: The T-SQL statement can be parsed, but causes the error at runtime. http://stackoverflow.com/questions/10166196/cannot-call-methods-on-nvarchar
Do you need your password? Powered by Blogger. here is the code for .aspx
I'm going to have to remember this myself! Can one bake a cake with a cooked egg instead of a raw one? This method eliminates the encoding problem you are seeing. MS SQL Server MS SQL Server 2005 MS SQL Server 2008 Rename and move Database and log to new volume in Exchange 2013/2016 Video by: Alan This video discusses moving either
Thanks for the quick help guys. FOR XML PATH command is altering data, trying to determine the cause. Word for "using technology inappropriately"? http://stackoverflow.com/questions/29853631/error-occured-cannot-call-methods-on-nvarchar Does this command perform any other data alteration I need to be aware of, does it handle unicode, etc.
Is the English word "ikebana" a suitable translation for "華道"? strogg 4-Dec-11 13:02pm You changed UserId to int? There will be another video to explain how to put the final p… MS Office Office 365 MS Access Advertise Here 773 members asked questions and received personalized solutions in the Privacy Policy.
I thought it might be the DATEADD call, but I took it out and still got the error. Change it to users –GraemeMiller Apr 15 '12 at 21:28 Chris is right :D –polohy Apr 15 '12 at 21:30 I still think you got that error Resolution: Errors of the Severity level 16 are generated by the user and can be fixed by the SQL Server user. Is it unethical to poorly translate an exam from Dutch to English and then present it to the English speaking students?
Versions: All versions of SQL Server. check over here up vote 0 down vote favorite Hi guys I have written this stored procedure but there is an error in it that says Cannot call methods on nvarchar T-SQL code: ALTER asked 4 years ago viewed 4860 times active 4 years ago Related 0nvarchar(max) slowing stored procedure4Getting an error converting data type nvarchar to int from application (C#) but not when running what was I going to say again?
What is the simplest way to put some text at the beginning of a line and to put some text at the center of the same line? All Forums Development Tools ASP.NET cannot call on method nvarchar Reply to Topic Printer Friendly Author Topic spshindagi Starting Member India 11 Posts Posted-01/24/2008: 02:39:20 the following is This statement works: SelectSQL = "SELECT user_id, user_name, user_surname, user_code FROM Users WHERE user_group = '" + drop.SelectedItem.Value + "'"; however, this line throws "Cannot call methods on nvarchar." SelectSQL = his comment is here Is it acceptable to ask an unknown professor for help in a related field during his office hours?
Example(s): DECLARE @xml varchar(MAX); SET @xml = '
I changed one method signature and broke 25,000 other classes.
Teenage daughter refusing to go to school Without opening the PHB, is there a way to know if it's a particular printing? Description: This error message appears when you try to call an invalid method for a data type. no return value either-2error occured : "Cannot call methods on nvarchar." Hot Network Questions Can I get a dual entry Schengen visa for tourism purpose for me and my wife? All Rights Reserved 4281 Express Lane, Suite L7710, Sarasota, FL 34238, Software Reviews | Book Reviews | FAQs | Tips | Articles | Performance Tuning | Audit | BI | Clustering
Why was Susan treated so unkindly? Our new SQL Server Forums are live! We've restricted the ability to create new threads on these forums. http://mobyleapps.com/cannot-call/cannot-call-methods-on-dialog-prior-to-initialization-attempted-to-call-method-39-destroy-39.html If a question is poorly phrased then either ask for clarification, ignore it, or edit the question and fix the problem.
How can I avoid being chastised for a project I inherited which was already buggy, but I was told to add features instead of fixing it? Your Email This email is in use. Post #914911 mister.magoomister.magoo Posted Monday, May 3, 2010 4:18 PM SSCrazy Group: General Forum Members Last Login: 2 days ago @ 10:30 AM Points: 2,184, Visits: 7,725 It sounds like you You may download attachments.
Wait... Optional Password I have read and agree to the Terms of Service and Privacy Policy Please subscribe me to the CodeProject newsletters Submit your solution! Join them; it only takes a minute: Sign up Cannot call methods on nvarchar up vote 3 down vote favorite I have strange problem with simple SELECT statement, which I use Actual meaning of 'After all' How can I avoid being chastised for a project I inherited which was already buggy, but I was told to add features instead of fixing it?
Template images by luoman. Also try by putting [] brackets around column names ujjwal uniyal 5-Dec-11 1:15am ok sir, i will try this. Change it to another misspelling and you will probably get a different error. Come on over!
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Tuesday, September 4, 2007
14 comments:
- Kim said...
Horrifying! That better not be on my side of the mailbox!
- September 4, 2007 at 7:34 PM
- Carol Dunstan said...
(in my best Mick Dundee voice) you call that a spider? This is a spider http://i36.photobucket.com/albums/e36/luincaran/spider.jpg
I know many will agree with you, but spiders don't really creep me out (though I am concerned about the poisonous ones)
- September 4, 2007 at 8:48 PM
- Tandra said...
Carol that is UG-LY!!! Tarantula??? grek!!!!
- September 4, 2007 at 8:53 PM
- Patricia said...
save it for Halloween!
- September 4, 2007 at 10:45 PM
- ~* Tracy *~ said...
EEK!!! LOL you are too funny!!
- September 5, 2007 at 1:35 AM
- Charmaine (CharmWarm on SCS) said...
That would give me the creeps too! Yuck!
- September 5, 2007 at 6:23 AM
- Kristina Lewis said...
Is that a real spider?? Oh my! What kind is it?? I HATE spiders!!
- September 5, 2007 at 2:01 PM
- Tandra said...
Oh YAH its real!! lol I could barely stand to get 6" away for the pic!!
I think its *just* a garden spidey. Last year we had one that had yellow and blue legs. A few months back Sharon Harnist (I think) posted on the same looking spider, and someone said it was a garden spider. Still....
I used to watch that GIANT spider movie when I was a kid, where it eats th epeople of th etown. It was in B/W I think!! LOL Yah but Im NOT that old- reruns...lol
- September 5, 2007 at 2:13 PM
- Carol Dunstan said...
Tandra your spider looks like some type of orb weaver... but then we probably have very different spiders to you! The one i posted a photo of is a giant water spider from what we could tell, my sister took the photo and estimated it was bigger than a man's hand...
- September 5, 2007 at 6:49 PM
- StamperSharon said...
See? They are creepy BIG, aren't they? Ick! Guess they are not just native to Texas, LOL!
- September 5, 2007 at 7:08 PM
- Dana said...
*giggle* I LOVE spiders, and am so impressed with this close-up shot! Nice.
- September 11, 2007 at 1:04 PM
- Rose Ann said...
Creepy, but good for the garden..I hear. ;)
- September 14, 2007 at 8:25 AM
- Godelieve said...
Good to see that I'm not the only one who takes pictures of spiders ;)
- October 14, 2007 at 9:48 PM
- The Stampin' Soldier said...
We used to have a "pet" spider like this. We called it a "zipper spider".
- November 13, 2007 at 5:24 PM


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Advertising insertion into VOD content is the next revenue stream for the cable sector. Julian Clover speaks to Tandberg's Edward Allfrey.
Targeted advertising has been a conference theme for about the last 18-months and is now getting to the stage when you feel it just might be about to happen. The driver is on demand, which takes on significance on this side of the Atlantic where there has never been the relationship between advertisers, broadcasters and cable operators as has been the case in North America.
Sitting down with Edward Allfrey, Business Development Director, Cable, Tandberg Television, it became clear that operators are beginning to think about the next stage in the development of their services. Allfrey has seen the business from both sides, having spent ten years with Telewest, before moving to the technology provider.
The first step is to look at the content, which in Europe has been driven by the catch up TV services, BBC iPlayer, SVT Play and TVPi. Although one of the key problems has been in navigating the available content, and how best to make new discoveries, Allfrey does not believe that the older interfaces have been entirely unsuccessful. "It's a user experience that people understand, and for the older and tech savvy they know how to find content in their EPG, so you can see that ability to find more content. That's why we see on the on demand platform a much wider demographic than the 16-35 year olds which are the basis for streaming video in the internet."
Allfrey has seen how VOD has been able to lock in customers, not only to their current tiers, but encouraging them to trade up to a level that unlocks a wider selection of content. Then there is the targeted advertising, one of the current flavours of the months, driven by the US market where there is a history of cablenets being able to sell their own advertising airtime within the cable–only channels.
"The great thing about advertising in on demand is that you can target it. Each stream goes to an individual household, so you can then say its easy to insert an advertisement based on their postcode, so that has intrinsically more value than the linear ad," says Allfrey.
There is already a clear interest from the operators, highlighted by Virgin's 100,000 home trial in North London last October, which according to the findings also recorded a greater awareness of individual ads.
Virgin already has the volume that makes it worthwhile to run targeted advertisements, other operators such as UPC are not too far behind of the curve, and the Amsterdam-based operator also has a history of local ad insertion.
Mark your diary for targeted advertising to be found on many a booth at IBC 2009.


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Stop the World, I Want to Get Off
January = New Year = New Year's Resolutions.
For most people, it's not long before resolutions are broken. According to Benjamin Hardy, only 8% of people accomplish their New Year's Resolutions. We spend the rest of the year accepting our failure and just doing what we did before. So there goes another year.
So how do we stop the cycle?
Step By Step
The problem is that we resolve to do something, but we don't identify the steps necessary to achieve the resolution. We resolve to lose weight, but we still stop by the donut shop every Friday. We resolve to read a certain number of books in a year and then spend our evenings in front of the TV.
We know what we want, we just don't think about what we need to change to make it happen. In the book Mojo: How to Get It, How to Keep It, How to Get It Back If You Lose It, Marshall Goldsmith defined 3 ways to handle change:
- Change yourself.
- Change the situation.
- Maintain the status quo and wonder why things aren't working.
People who choose option 3 don't keep their resolutions. So we need to choose Curtain 1 or Curtain 2, Or sometimes both. How do we do that? By addressing habits.
You resolve to read more because you just couldn't find the time to read last year. If you watch less TV, then you'll have more time to read. So instead of making some generic resolution to read more, put a book near your favorite chair and move the remote to the other side of the room. Take a magazine with you when you are going to an appointment. Give yourself some fresh air by reading during lunch. You have opportunities to make small changes to make reading a habit.
Your Change Life Campaign will focus on changing the habits so that you can achieve your goals.


text:
The costs for treating a severe injury caused by an accident can be considerable. Extensive hospital treatments, long-term rehabilitation or a lifetime of continuous care may be necessary. Costs don't remain stagnant -- medical expenses for Chicago spinal cord injury patients increase year after year.
Medical mistakes Illinois doctors are ignoring
Chicago patients visit doctors with the expectation of an accurate diagnosis -- maybe not on the first visit but eventually. We trust doctors to get it right, but that doesn't happen as much as patients would like to believe. Misdiagnoses and delayed diagnoses are frequent occurrences, even for common illnesses.
Pinpointing surgical errors to prevent medical malpractice
In the medical community, as with other professions, problems must be acknowledged and identified before solutions can be effective. The risks of causing serious injuries or deaths are high when Chicago doctors and hospitals fail to change unsafe behavior patterns.
What Chicago parents don't know about dangerous toys
Chicago parents may assume all children's products sold to consumers are safe. After all, we have strong government rules set up to safeguard us against the sale of dangerous children's toys. As high as safety standards are in the U.S., some defective children's products slip into the marketplace.


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Infant, tests showed small hiatal hernia, fussy with food. Suggestion?
My 9 month old grandson is having eating issues. Most days it is a struggle to get him to take his bottle. Over the last week or so it has escalated. Yesterday he took only 18 oz. of formula. He currently is on neocate (sp?) after being on 5 other formulas. He will eat some strained foods but maybe one jar a day. He is a happy baby and extremely active. He crawls and walks along side furniture. He is very strong. His ped. said he couldn t help and suggested a gastroent.. After initial meeting, upper GI and endoscope all they have come up with is a small hiatal hernia . Have seen a feeding specialist with no luck. Worried about dehydration . Any suggestions?
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Sat, 21 Jul 2012
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User's Response
Hihi,Raw juices extracted from fresh fruits and vegetables are valuable in haitus
,and the patient should take these juices half an hour before each meal.carrot juice is specially bebeneficial as it has a very restorative effect,and is rich in
vitamin A
and calcium.it is an alkaline,food,which soothes the stomach.The foods which should be avoided are over-processed foods like white bread and sugar,cakes and biscuits,rice puddings and overcooked vegetables. Take care
Answered: Sun, 22 Jul 2012
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Hihi,Raw juices extracted from fresh fruits and vegetables are valuable in haitus hernia,and the patient should take these juices half an hour before each meal.carrot juice is specially bebeneficial as it has a very restorative effect,and is rich in vitamin A and calcium.it is an alkaline,food,which soothes the stomach.The foods which should be avoided are over-processed foods like white bread and sugar,cakes and biscuits,rice puddings and overcooked vegetables. Take care
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GT Prime, GT Saber, TG Blaster & TG Skywarp Released
Posted 30 May 2013 - 02:02 PM
Posted 01 June 2013 - 07:13 PM
So take that as you will (as it seems it could mean either same old guys in new bodies or new guys based on old guys to me).
#63 (Deactivated) XeroWarlock
Posted 11 June 2013 - 11:36 AM
I kinda feel like I've been ripped off a bit. GT-R Prime I was terrified would break just trying to get the hood to collapse. This thing would not budge.
And the legs on both of these... gah! Who was smoking what when they figured that was a good idea?


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Review: Foxy Shazam at Town Ballroom
Since 2004, this Cincinnati-based band has been rocking out venues and entertaining ever-growing audiences with their jazzy-rock alternative beats.
On June 25, Foxy Shazam took over Buffalo's Town Ballroom, drawing in fans of all shapes and sizes for the same reason: to listen to Foxy's new and old music, while watching one helluva sildenafil farmacias show.
Foxy isn't the kind of band that takes their places, strums their strings, beats their drums, sings their notes and then calls it a night. No. Schuyler Vaughn White does handstands while playing his keyboard, Alex Nauth dances all over the stage while playing his trumpet, Daisy Caplan and Loren Daniel Turne hold guitars above their heads, Aaron McVeigh bashes his symbols off of Eric Sean Nally head who was endlessly flipping and rolling and jumping around the stage while singing songs from all of their albums.
It wasn't just a concert, it was a full-blown show. And the crowd in the pit was just as amped up as the band was.
They started their set with "Gonzo", the single off of their newest album with the same title. Starting out with their most recent hit got everyone's attention right off the bat, and they kept the show going with energized hits from their 2014 album.
As they concluded that set, they
announced that it was just an intermission. And sure enough, Foxy swarmed the stage once again, with new outfits and whole new vibe, throwing the audience back to their last album "The Church of Rock and Roll" with
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No one can say that this show lacked variety of any sorts.
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Even those who may not have been die-hard fans, or maybe had not heard their music before at all, were definitely entertained throughout Foxy's performance in June at The Town Ballroom.
If given the chance to witness these guys live, it's definitely worth the money.


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Marybeth Shea's curator insight, December 20, 2012 9:44 AM
Another plant ally for human health, bolstering case for plant conservation as insurance option for human development.
Meristemi's comment, January 21, 2013 4:21 PM
Can't decide if the ignorance of botany is worse than nationalism, or not.
Christian Allié's curator insight, January 10, 2014 10:08 AM
...... These 7 extreme flowers include the world's largest, smallest, stinkiest and most dangerous. Stunning examples of the incredibly unexpected wonders that nature can serve up, the world's most bizarre blooms entice, amaze and disgust.................
Marybeth Shea's curator insight, December 26, 2012 5:54 PM
Fabulous idea. Let your small plot be a move toward biodiversity conservation.
Marybeth Shea's curator insight, December 20, 2012 9:46 AM
Bacteria farms for ambergris? A whale of an idea :). Bacteria are a little-understood natural resource for many uses.


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I May Be Watching Too Much TV
December 08, 2009
So I'm watching young beautiful people fling themselves around on a stage and I had an inspiration for a new reality show combining "So You Think You Can Dance" and "Project Runway". You could call it -- wait for it -- So You Think You Have Pants! It's a good thing my Caps played last night instead of tonight, because I hate choosing between fast young men skating and beautiful young people dancing. It's my... Read more →


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By Dennis Polkow
When French pianist Pierre-Laurent Aimard came to Chicago in 1986, it was as a member of Pierre Boulez's l'Ensemble InterContemporain for a week of performances. At that time, Aimard had already been playing with the Ensemble since its inception a decade earlier.
"It was such an exciting time," Aimard recalls. "Boulez had been active abroad and was living in Germany but the moment he came back to France, there was so much anticipation." Boulez did not disappoint: he founded the Paris-based IRCAM, the Institute for Research and Coordination in Acoustics and Music, with the goal of enlarging the domain of materials used for music. That goal was embraced by musicians of all genres and even brought Frank Zappa to Boulez.
When Boulez himself asked Aimard to join IRCAM's new resident ensemble, "it was a privilege, and I thought I would be there for a couple of years." He would remain for eighteen years, before finally setting off to have a career of his own in 1994. "I was overwhelmed by the power of his artistry, of his musicianship, his fabulous intellect, his work ethic and the commitment that he gave to all of the pieces he was serving. It was a happy eighteen years."
In celebration of Boulez's ninetieth birthday on March 26, Aimard and Yugoslavian-born pianist Tamara Stefanovich—who was a student of Aimard's seventeen years ago and now has a career in her own right—will present a recital of the complete piano works of Boulez, music that Aimard has lived with most of his life. "I began working on the Second Sonata when I was twelve to thirteen years old," says Aimard. "I tried to analyze it, I was fascinated. In his early years, Boulez was a pianist and his most important early works are all piano pieces. When he became a conductor, his most important pieces became orchestral pieces."
The piano pieces, Aimard feels, show Boulez emerging as the most important musical voice after the Second World War. "The world was being reconstructed and composers were looking for new expressions. There is such richness in this music, always new depths and meaning. We will perform the pieces chronologically and talk a little about their significance." The recital will close with Aimard and Stefanovich performing Boulez's monumental 1961 composition, "Structures II," for two pianos together. March 15, 3pm, Orchestra Hall, 220 South Michigan, (312)294-3000.
Muti Mourns Patner
Much has been written about the sudden death of arts critic and broadcaster Andrew Patner, but Chicago Symphony Orchestra music director Riccardo Muti uniquely honored his memory by co-dedicating his recent, extraordinarily moving performances of the Mozart "Requiem" to Patner. There will also be a public memorial service at Orchestra Hall on March 18. If these gestures seem unprecedented, so was the friendship that Muti and Patner shared.
"I wanted to do this because for me, the loss of Andrew has been something very painful," Muti explains. "He was one of the very few persons in this city with whom I could exchange ideas and have a conversation full of humor and have a good laugh, but also speak seriously about art, about society, about philosophy, about theology. This is becoming very rare in the world.
"When I was a young boy, there was no television in Italy so on Saturday or Sunday afternoons, we would speak with our teachers who were in the park or outside the school. With Andrew, we had this possibility and I wanted to hear his ideas—not necessarily to agree with them, but they were always profound. So, the dedication was not just an act.
"I lost a brother as well this year. When I go back to the village where I grew up, still now, I go back to the cemetery because I like to see all of the inscriptions on the graves. One that always impressed me was where a father wrote on the grave of his son who died very young, 'It is not life that is going to die, it is death that is going to die.' It's very profound. It means that our life here, our physical life is the real inferno. The father was telling his son, 'You are leaving the inferno here and going back.'
"In Italy, especially as Southern Italians, we grew up with a sense of death, and when I conduct a 'Requiem,' I try to convey this concept of this very tight relationship between the living and the dead. When we do this music, it is like the entire world of our friends and our relatives who are dead are together with us." March 18, 7pm, Orchestra Hall, 220 South Michigan, (312)294-3000.


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Wright Words: Bestselling author shares theory — and his cellphone number
netmaster Farmington, UTFeb. 20, 2013 10:30 p.m.
I used to work for a national magazine and had a measure of fame because I was well-known among those whose hobbies or jobs were connected with the topic that my magazine covered. I tried to remain accessible and never regretted it. Some of the people who contacted me because I had knowledge they desired or simply because they were enamored by a celebrity in their eyes became and have remained close friends. And they are true friends because I'm no longer famous (and don't wish to be) and yet they are there for me when I need somebody to lend me an ear or a hand. Early in my career I once told a man that I did not want to sign an autograph on an article that I had written, but my father-in-law told me that such a small gesture would be appreciated, even if I didn't consider my autograph worthwhile or my work particularly special or worthy of praise. From that time on I never refused to spend a few minutes with anybody who requested my time. And the rewards have been ample. Aloof celebrities are missing out on those rewards.
rogerdpack2 Orem, UTFeb. 19, 2013 1:53 p.m.
wow this guy has guts.
SLC gal Salt Lake City, UTFeb. 19, 2013 8:23 a.m.
I think sharing your cell is above and beyond the call of duty, but I think that's awesome you're willing to connect to your fans on such a personal level.
DarrellL SAN BERNARDINO, CAFeb. 19, 2013 7:52 a.m.
Thank you, Mr. Wright!