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Transforming growth factor (TGF) β1 regulates cell migration of non-neural cells. Hence, two hypotheses were tested: (i) that TGFβ1 affects cell migration and the expression of associated adhesion proteins in developing cortex; and (ii) that these effects are antagonized by ethanol. The effects of TGFβ1 (2.5–40 ng/ml) and ethanol (400 mg/dl) on cell migration were examined in organotypic cultures from fetal rat brains. Migration was determined by tracing the movement of cells pulse-labeled with bromodeoxyuridine. Cell migration was altered by TGFβ1 in a concentration-dependent manner: at low concentrations, cell migration was promoted whereas at high concentrations TGFβ1 impeded migration. Ethanol treatment alone reduced the rate of migration. Interestingly, the rate of cell migration in slices treated with both TGFβ1 and ethanol was the same as that in untreated cultures. The expression of cell adhesion proteins (nCAM, integrin α3, αv and β1) was differentially effected by TGFβ1 and/or ethanol. TGFβ1 increased the expression of these adhesion proteins in a progressive, concentration-dependent manner. Likewise, ethanol also increased adhesion protein expression, however, combined TGFβ1 and ethanol treatment reduced expression. Collectively, the data show that TGFβ1 alters cell migration in the developing cortex and that the TGFβ1 system is a target of ethanol toxicity.
Development of the cerebral cortex from a simple, embryonic neuroepithelium to a well organized laminar structure requires proper migration of post-mitotic neurons. Migration is a highly regulated process following an orderly inside-to-outside sequence (Angevine and Sidman, 1961; Berry and Rogers, 1965; Rakic, 1972, 1974; Miller and Nowakowski, 1988; Rakic et al., 1994). The order of this process depends upon the coordination of migrating neurons, radial glia, and proteinaceous mediators in the microenvironment.
Cellular interactions important for migration are mediated through multiple groups of proteins, including cell adhesion proteins (CAPs) and growth factors. CAPs, such as neural cell adhesion molecule (nCAM), L1, astrotactin, and integrins, mediate appropriate attachment in the extracellular environment and transduce signals to cytoskeletal elements important for cell motility (Miura et al., 1992; Zheng et al., 1996; Lambert de Rouvroit and Goffinet, 2001; Hatten, 2002; Nadarajah and Parnavelas, 2002; Sobeih and Corfas, 2002; Schmid and Anton, 2003). One growth factor that affects cell migration is transforming growth factor (TGF) β1. For example, TGFβ1 can promote the migration of a variety non-neural cells including osteoclasts (Pilkington et al., 2001), immune (Wahl et al., 1993; Olsson et al., 2000; Olsson et al., 2001), and endothelial cells (Enenstein et al., 1992). TGFβ1 is also involved in tumorgenic cell migration, particularly that of neural-derived gliomas and neuroblastomas (Paulus et al., 1995; Tsuzuki et al., 1998; Platten et al., 2000; Platten et al., 2001).
The distributions of ligands and receptors in the developing cortex in vivo are consistent with the notion that TGFβ is involved in neuronal migration. TGFβ ligands (TGFβ1 and TGFβ2) are expressed by neurons and radial glia, respectively (Flanders et al., 1991; Miller, 2003). TGFβ receptor type 1 (TGFβIr) mRNA is found in the intermediate zone (IZ) of the developing mouse cortex (Tomoda et al., 1996). Prenatally, both TGFβIr and TGFβ receptor type 2 (TGFβIIr) protein are expressed by cells in the proliferative areas. Perinatally, TGFβIr is expressed along radial glial fibers (Miller, 2003). Collectively, these studies provide compelling evidence that the TGFβ system is involved in neuronal migration in the developing brain.
TGFβ1 can influence cell migration by modulating the expression of the CAPs that mediate attachment and promote motility. Two families of adhesion proteins, integrins and cell adhesion molecules (CAMs), are upregulated by TGFβ in numerous cell types (Ignotz and Massagué, 1987; Roberts et al., 1988; Heino et al., 1989; Ignotz et al., 1989; Roubin et al., 1990; Perides et al., 1994; Stewart et al., 1997; Luo and Miller, 1999a; Miller and Luo, 2002). Integrins are heterodimeric proteins that act as essential links between proteins in the extracellular environment and the cytoskeleton (van der Flier and Sonnenberg, 2001). CAMs, e.g. nCAM and L1, mediate cellular attachment through homophilic interactions or binding with CAPs (Ronn et al., 1998; Crossin and Krushel, 2000). Studies of immature neurons and neuroblastoma cells show that TGFβ1 up-regulates expression of nCAM (Luo and Miller, 1999a; Miller and Luo, 2002), a protein intimately involved in neuronal migration (Chuong et al., 1987; Miura et al., 1992).
Migrating neurons are targets of ethanol toxicity. Brains of human fetuses exposed to alcohol in utero exhibit defects such as neuroglial heterotopias and ectopic cell clusters (Clarren et al., 1978). Studies of animal models provide insight into possible mechanisms underlying such defects. In the neocortex, late-generated neurons intended for superficial cortical layers can be found ectopically in deeper layers of cortex (Miller, 1993). A potential cause of this migrational error is the disruption of the radial glial scaffold upon which some neurons migrate because the radial glia prematurely differentiate into astrocytes (Shetty and Phillips, 1992; Miller and Robertson, 1993). Alternatively, neurons destined for the cortical plate (CP) can form heterotopic clusters beyond the limits of the cerebral wall (Clarren et al., 1978; Kotkoskie and Norton, 1988; Komatsu et al., 2001). Ethanol can disrupt signals that prompt post-mitotic neurons to migrate from the proliferative zones and reduces the rate of neuronal migration. Ethanol exposure, however, does not reduce the total number of migrating cells within the cerebral wall (Miller, 1993). Taken together, these data suggest that ethanol affects the initiation, maintenance, and completion of neuronal migration.
Ethanol interferes with the normal expression and activity of TGFβ1. In vivo, prenatal ethanol exposure alters the amount of both TGFβ ligand and receptor in the developing cortex (Miller, 2003). Notably, ethanol severely reduces the expression TGFβIr in radial glia. In vitro, ethanol blocks TGFβ1-dependent increases in nCAM expression by B104 neuroblastoma cells (Luo and Miller, 1999a). Furthermore, treatment of primary cultured cortical neurons with either TGFβ1 or ethanol increases nCAM expression (Miller and Luo, 2002). In contrast, combined treatment with TGFβ1 and ethanol attenuates nCAM expression. Thus, it appears that TGFβ1 and ethanol affect cellular changes via similar, yet unidentified, mechanisms. Functional studies of neuroblastoma cells support this conclusion. TGFβ1 and ethanol are anti-proliferative agents that initiate the same signal transduction mechanisms (Luo and Miller, 1999a,b). It is noteworthy that ethanol also affects the function of L1, by reducing L1-mediated cell–cell adhesion (Charness et al., 1994; Ramanathan et al., 1996; Wilkemeyer et al., 1999, 2002).
The present study tests the hypotheses that TGFβ1 modulates cell migration in the developing cerebral cortex and that ethanol disrupts the action of TGFβ1. Two primary goals of this study are (i) to determine whether TGFβ1 influences cell migration and CAP expression in the developing cerebral cortex; and (ii) to assess whether ethanol affects the action of exogenously applied TGFβ1.
Materials and Methods
Organotypic Slice Cultures
Pregnant Sprague–Dawley rats (Taconic, Germantown, NY) were anesthetized on gestational (G) day17 with a cocktail of ketamine (10 mg/kg) and xylazine (1.0 mg/kg). Embryos were collected and their brains were removed. Forebrains were dissected and cut coronally into 300 µm slices using a MacIlwain Tissue Chopper (Mickle Lab Engineering, Gomshell, UK). All slices were taken from the same rostrocaudal level in the brain that contained presumptive parietal cortex. Approximately six slice hemispheres were obtained per fetal brain. Slices from a single litter were pooled and carefully arranged on Millicell filter inserts with 0.40 µm pores; five or six slices per insert (Millipore, Bedford, MA).
Inserts were placed in 35 mm culture dishes (Falcon, Lincoln Park, NY) containing 20% fetal bovine serum (FBS), Minimal Essential Media with Hanks salts, 200 mM glutamine (Gibco, Carlsbad, CA), 25 mM HEPES, 100 mM dextrose, 25 mM KCl, 100 µM penicillin/streptomycin (Gibco), and 12.5 µM fungizone (Gibco). HEPES was added to further stabilize the pH of the medium during regular movements in and out of the incubator necessitated by the ethanol and 5-bromo-2-deoxyuridine (BrdU; Sigma, St Louis, MO) treatments. Cultures were incubated at 37°C with 95% O2/5.0% CO2. Slices remained in the FBS-laced medium for 2–4 h, after which the slices were incubated in a FBS-free medium containing TGFβ1 (0, 2.5, 5, 10, 20 or 40 ng/ml), ethanol (100, 200, 400 mg/dl), or TGFβ1 (10 ng/ml) and ethanol (400 mg/dl).
An enclosed chamber method was used to preserve a stable concentration of ethanol in the medium (Adickes et al., 1988; Luo and Miller, 1997). Cultures were suspended over a water bath containing 100, 200 or 400 mg/dl ethanol, depending on treatment. The chambers were sealed and CO2 was added to the chambers to maintain a concentration of 5.0% CO2.
Bromodeoxyuridine Labeling and Immunohistochemistry
Following 10 h treatment with TGFβ1 and/or ethanol (designated as time 0 hr; t0), slices were exposed to 0.0040% BrdU for 1 h (Luo and Miller, 1997; Haydar et al., 1999; Jacobs and Miller, 2000). After the 1 h exposure (t1), the BrdU-containing medium was removed and replaced with fresh, serum-free medium containing no BrdU. At t1, t6 or t12, slices were fixed in 4.0% paraformaldehyde for 30 min at room temperature. Slices were processed for cryosectioning and cut in 15 µm sections.
Cells that incorporated BrdU were visualized immunohistochemically (Miller and Nowakowski, 1988; Luo and Miller, 1997; Jacobs and Miller, 2000). Following a rinse with 0.10 M phosphate buffered saline (PBS) rinse, endogenous peroxidase activity in the sections was quenched with a wash in 3.0% H2O2 for 5 min. Sections were incubated in 2.0 N HCl for 30 min followed by a 1 min rinse in dH2O. Sections were incubated overnight at 4°C with an anti-BrdU antibody (Becton-Dickinsen, San Diego, CA) diluted 1:30 in PBS with 0.75% Triton X-100 and 10% goat serum. After a several rinses with PBS, sections were incubated in biotintylated goat anti-mouse antibody (Vector Laboratories, Burlingame, CA) diluted 1:400 in PBS for 1 h. Elite Vectastain ABC and DAB kits (Vector) were used to detect BrdU-labeled cells.
Some sections were processed immunohistochemically for the expression of nCAM. A procedure similar to that described above was followed with the following differences: (i) the sections were not treated with HCl; and (ii) the primary antibody was a mouse anti-nCAM antibody (Sigma) diluted 1:1000 in PBS.
Various controls for the immunolabeling were performed. For nCAM and BrdU labeling, the immunohistochemical procedure was performed without incubating the sections with a primary antibody. A second control for the BrdU studies included immunohistochemical processing of sections from slices not exposed to BrdU. The results of these controls were consistently negative.
Cerebral Wall Collection and Immunoblotting
Changes in the expression of CAPs in the cerebral wall were detected using a quantitative immunoblotting method. Slices were prepared and treated as described (see 'Organotypic slice cultures') and collected at t6 and t12. The section of cerebral wall used in the immunoblot analyses was limited laterally by the lateral extent of the ventricle and medially by the midpoint of the ventricle (Fig. 1A; dashed lines and arrows). The cerebral walls of eight slices from each treatment condition were collected and pooled.
The tissue was placed in a lysis buffer (1.0% Nonidet P-40, 0.50% deoxycholic acid, 0.010% sodium dodecylsulfate (SDS), Complete Mini protease inhibitor cocktail tablets (1 tablet per 10 ml buffer; Roche, Indianapolis, IN) in 0.010 M PBS), homogenized, and spun at 10,000 r.p.m. for 10 min. Only the supernatant was used for protein analysis. Pilot studies performed on pellet fractions consistently showed that no signal was detectable. The protein content of the supernatant was determined (Bradford, 1976). An aliquot of the supernatant containing 40 µg/ml protein was combined with electrophoresis sample buffer (300 mM Tris–HCl, 50% glycerol, 5.0% SDS, 0.025% bromophenol blue, w/wo 250 mM β-mercaptoethanol).
Samples were loaded on a 10% SDS–polyacrylamide gel, separated by electrophoresis, and transferred to nitrocellulose membranes. Non-specific immunoreactivity on the membranes was blocked for 2 h in 5.0% non-fat dehydrated milk (NFDM). Blots were then probed overnight at 4°C with one of the following: mouse monoclonal anti-nCAM antibody (1:1000 in PBS; Sigma) diluted in 2.5% NFDM, rabbit-polyclonal anti-integrin α3 antibody (1:200 in PBS; Chemicon, Temecula, CA), mouse monoclonal anti-integrin αv antibody (1:500 in PBS; Pharmigen, San Diego, CA), and rabbit polyclonal anti-integrin β1 antibody (1:200 in PBS; Santa Cruz, Santa Cruz, CA). Subsequently, the membranes were incubated for 30 min with horseradish peroxidase-linked anti-mouse or anti-rabbit secondary antibody. Tagged protein bands were detected using a chemiluminescent detection reagent (Amersham, Piscataway, NJ). In most cases, the membrane was stripped of immunolabel and re-probed with an anti-actin antibody as a loading control (1:1500; Sigma).
Two standards were loaded onto each gel. One was an internal standard, which was lysate of pooled cortices from a litter of 19-day-old rat fetuses. Two samples of this internal standard were loaded onto each gel. Rainbow protein standards (Amersham) were also loaded onto each gel to determine the molecular weight of the immunolabeled protein.
The rate of migration was calculated by determining the mean change in the mean position of the ten outermost BrdU-labeled cells between t6 and t12. The analysis focused on a 200 µm wide strip of the lateral cerebral wall (Fig. 1A; boxed region). The mean distance of the ten BrdU-labeled cells farthest from the ventricular surface was determined with the Bioquant Image Analysis System (R&M Biometrics, Nashville, TN). Measurements were taken from 3–5 slices from a single litter for each timepoint and treatment condition. A minimum of four litters (n ≥ 4) was used to represent each datum.
Densitometric analysis of the immunoblots was performed using an Image Station (Kodak, Rochester, NY). Data were corrected for loading variations within each blot using the amount of actin expression as a standard. Inter-blot variation was addressed by normalizing the data against the signal detected in the internal standard (Mooney and Miller, 2001).
Differences among treatment groups were assessed with an analysis of variance (ANOVA). In cases in which statistically significant (P < 0.05) differences were detected, specific differences (e.g. in time or treatment) were examined using individual t-tests.
Cell Migration in Organotypic Slice Cultures
In order to identify a discrete population of cells within the cerebral wall, slices were exposed to a one h pulse of BrdU (t0 to t1). At t1, BrdU-positive cells in the untreated (control) slices were confined to the ventricular zone (VZ) and subventricular zone (SZ) (Fig. 1B). This is consistent with the understandings (i) that BrdU is incorporated into cells passing through the S-phase and (ii) that cycling neural cells are distributed in the VZ and SZ (Angevine and Sidman, 1961; Hinds and Ruffett, 1971; Rakic, 1974; Miller and Nowakowski, 1988; Nowakowski et al., 1989). Six hours after introducing the BrdU (t6), most BrdU-labeled cells were in the proliferative zones, but some were distributed in the deep IZ (Fig. 1C). Presumably, this change resulted from the migration of BrdU-positive cells from the proliferative zones into the IZ. At t12, additional BrdU-labeled cells were in the IZ and they were located farther into the IZ (Fig. 1G).
To quantify the change in the distribution of BrdU-positive cells in the untreated slices, the distances of the 10 outermost labeled cells from the ventricular surface at t1, t6 and t12 were measured. At t6, the outermost 10 cells were significantly (P < 0.05) farther from the ventricular surface as compared with t1 (Figs 2 and 3A). These findings indicated that the cells had commenced their migrations by t6. By t12, the 10 outermost cells were distributed farther from the ventricular surface (Fig. 3A). The mean change in the position of the outermost cells between t6 and t12 was indicative of the rate of migration. Accordingly, the mean rate of migration of cells in untreated slices was 6.1 µm/h (Fig. 3B).
TGFβ1-promoted Migration of Post-mitotic Cells
At t1 (data not shown) and t6 (Fig. 1C,D), the distribution of the outermost BrdU-positive cells in the slices treated with TGFβ1 (10 ng/ml) was comparable to that in the untreated preparations. By t12, more BrdU-positive cells in the TGFβ1-treated slices were distributed in the IZ than at t6 and they were located more superficially (i.e. farther from the ventricular surface) as compared with untreated conditions (Fig. 1G,H). Thus, by t12, cells in the TGFβ1-treated slices had migrated farther than cells in untreated slices.
The effect of TGFβ1 (10 ng/ml) on cell migration was quantified. At t1 and t6, the outermost cells in TGFβ1-treated slices were at a similar mean distance from the ventricular surface as in untreated slices (Figs 2 and 3A). The mean position of the ten outermost cells significantly (P < 0.05) differed in TGFβ1-treated slices at t1 and t6 (Figs 2 and 3A). At t12, the outermost population in TGFβ1-treated slices was significantly (P < 0.05) farther from the ventricular surface than cohorts in untreated slices (Fig 3C). The mean rate of migration was significantly (P < 0.05) increased (74%) by treatment with TGFβ1 (Fig. 3B).
TGFβ1 affected cell migration in a concentration-dependent manner [Fig. 3B; F(7,31) = 13.038; P < 0.001]. At low concentrations (i.e. 2.5 and 5.0 ng/ml), TGFβ1 treatment induced a modest increase in the rate of migration (32 and 36%, respectively), however, these differences were not statistically significant (Fig. 4B). The rate of migration nearly doubled in slices treated with 10 ng/ml TGFβ1 (as compared with the rate of migration in untreated slices); this increase was significant (P < 0.05). At concentrations above 10 ng/ml, however, no such increase was evident. In slices treated with high amounts of TGFβ1 (40 ng/ml), the rate of migration was significantly (P < 0.05) slower than it was in either untreated slices or slices treated with 10 ng/ml of TGFβ1 (Fig. 4B). Thus, TGFβ1 did not always facilitate cell migration; indeed, at high concentrations, TGFβ1 inhibited migration.
Ethanol Impeded Cell Migration
The effect of ethanol (400 mg/dl) on the spatiotemporal change in the location of BrdU-labeled cells was determined. At t1, BrdU-positive cells in the ethanol-treated slices were in the proliferative zones, a pattern that was indistinguishable from that in the untreated and TGFβ1-treated preparations (data not shown). Unlike the untreated and TGFβ1-treated slices at t6, in the ethanol-treated slices few BrdU-labeled cells were outside the proliferative zones and were distributed closer to the ventricular surface (Fig. 1E). At t12, more cells were evident in the IZ, however, they were located deeper in the IZ than were cohorts in untreated slices (Fig. 1G,I). This implies that ethanol indeed affects cell migration.
Quantitative analysis of the period from t1 to t12 provided key information about the effects of ethanol on cell migration. At t1 the outermost BrdU-positive cells in the ethanol-treated slices were positioned similarly to those in the untreated and TGFβ1-treated preparations (Fig. 2). Moreover, there was a significant (P < 0.05) change in the mean distance of the ten outermost cells in the ethanol-treated slices between t1 and t6 (Figs 2 and 3A). That is, as in the untreated and TGFβ1-treated preparations, cell migration had commenced by t6. Further analysis showed that the outermost BrdU positive cells at t6 and t12 in ethanol-treated slices were significantly (P < 0.05) closer to the ventricular surface than were cohorts in the untreated slices (Fig. 3A). In fact, the rate of migration of the outermost ten BrdU-positive cells was significantly (P < 0.05) reduced (43%) by ethanol (400 mg/dl; Fig. 3B). Treatment with lower amounts of ethanol (100 or 200 mg/dl) also attenuated the rate of migration (data not shown). Both concentrations reduced the rate of migration 21–26% relative to controls.
The outermost BrdU-positive cells in slices co-treated with TGFβ1 (10ng/ml) and ethanol (400 mg/dl) were distributed at similar positions in the cerebral wall as those in untreated slices (Fig. 1C,F,G,J). The mean position of the outermost 10 cells in co-treated slices was not significantly different than in untreated preparations at both t6 and t12 (Fig. 3A). Furthermore, the mean rate of migration in TGFβ1-ethanol-treated slices was not significantly different than untreated cultures (Fig. 3B). Interestingly, the mean rate of migration in co-treated slices was significantly (P < 0.05) different than either conditions alone, being less than TGFβ1 but greater than ethanol (Fig. 3B). This suggests that ethanol disrupts TGFβ1-dependent changes in cell migration.
TGFβ1 Increased CAP Expression
CAPs, such as nCAM and integrin subunits α3, αv and β1, are important for cell migration in the developing brain (Chuong et al., 1987; Georges-Labouesse et al., 1998; Anton et al., 1999; Graus-Porta et al., 2001). Thus, cortical slices were examined for TGFβ1-dependent changes in CAP expression. Immunoblotting methods were used to determine whether TGFβ1 affected nCAM expression. At t6, concentrations of TGFβ1 (2.5, 10 and 40 ng/ml) significantly [F(5,14) = 27.934; P < 0.001] increased expression of nCAM (Fig. 5). The greatest increase in nCAM expression was detected in cultures treated with the highest concentration of TGFβ1. Some change in nCAM expression occurred between t6 and t12, though it was not statistically significant. On the other hand, by t12, the expression of nCAM in TGFβ1-treated slices was no longer significantly higher than untreated at t12. This was because at t12, nCAM expression in untreated cultures had risen significantly (P < 0.05) as compared with the amount of expression at t6.
The distribution of nCAM expression in the cerebral wall was also examined immunohistochemically. In untreated slices, nCAM expression was highest in the IZ and CP, though some labeling was evident in the proliferative areas (Fig. 6A). nCAM immunolabeling was particularly notable on the cell membranes where dark puncta outlined cell bodies (Fig. 6G). Changes in nCAM labeling were evident in slices treated with TGFβ1. These changes appeared to be concentration-dependent. Low concentrations of TGFβ1 (2.5 and 10 ng/ml) induced subtle changes in nCAM expression, most notable around the outside of the cell body (Fig. 6B,C,H,I). Treatment with high concentrations of TGFβ1 (40 ng/ml), however, induced substantial changes (Fig. 6D). Dark, often large patches of immunostained membrane were frequently evident (Fig. 6J).
TGFβ1 also altered expression of the integrin subunits α3, αv and β1. The pattern of integrin α3 expression was similar to that for nCAM. At t6, integrin α3 expression was increased significantly [F(5,19) = 16.347; P < 0.001] by TGFβ1 with treatments of 40 ng/ml having the largest effect (Fig. 7). Whereas integrin α3 expression in the TGFβ1-treated slices was not significantly different between t6 and t12, expression in the untreated conditions increased significantly (P < 0.05) by t12. Thus, α3 expression in the untreated slices did not vary significantly from the TGFβ1-treated slices at t12. Unlike nCAM expression, integrin αv expression only increased significantly [F(5,16) = 3.811; P < 0.05; F(5,17) = 5.771; P < 0.05; t6 and t12, respectively] following treatment with a high concentration of TGFβ1 (Fig. 8). TGFβ1 treatment also significantly [F(5,16) = 28.945; P < 0.001] increased the expression of integrin β1 at t6 with all concentrations tested (Fig. 9). On the other hand, at t12 only treatment with a high concentration of TGFβ1 significantly [F(5,14) = 7.175; P < 0.01] increased integrin β1 expression.
Ethanol Affected CAP Expression
To determine whether ethanol affected CAP expression, immunoblot and immunohistochemical analysis were used. Ethanol (400 mg/dl) significantly (P < 0.05) increased expression of nCAM at t6 (Fig. 5). Lower concentrations of ethanol (100 and 200mg/dl) likewise increased nCAM expression (data not shown). The increased expression of nCAM expression was maintained between t6 and t12. Interestingly, the ethanol-induced increase in nCAM expression was similar to that detected in slices treated with a high concentration of TGFβ1 (Fig. 5). The changes in nCAM expression were paralleled by immunohistochemical changes. nCAM immunolabeling was prominent in slices treated with ethanol, particularly in the IZ and CP (Fig. 6E). Intense regions of staining were commonly found around the outside of the cell body (Fig. 6K).
Like nCAM, expression of the integrin proteins was affected by ethanol. Ethanol significantly (P < 0.05) increased the expression of integrins α3, αv and β1 at t6 (Figs 7–9). In addition, the expression of integrin αv and integrin β1 was significantly (P < 0.05) greater in ethanol-treated slices than in untreated preparations at t12 (Figs 8 and 9).
Slices treated with both TGFβ1 (10 ng/ml) and ethanol (400 mg/dl) were examined for changes in nCAM expression. These preparations were subjected to three comparisons: with untreated slices and with cultures treated with TGFβ1 or ethanol. Co-treatment significantly (P < 0.05) increased the amount of nCAM at t6 relative to the amount of expression detected in untreated slices (Fig. 5). nCAM expression in slices co-treated with TGFβ1 and ethanol was not significantly different from that of slices treated with TGFβ1 alone. In contrast, TGFβ1-ethanol co-treatment significantly (P < 0.05) reduced nCAM expression at t6 and t12 re slices treated with ethanol alone.
In the co-treated slices, nCAM was expressed by cells in the IZ and CP. That is, the pattern of nCAM immunolabeling within the cerebral wall of co-treated slices was the same as that in slices treated with TGFβ1, ethanol, or neither substance. On the other hand, the intensity of the immunolabel appeared to be affected by co-treatment. nCAM expression in the IZ of co-treated appeared to be heterogeneous as there were cells with the intense staining characteristic of ethanol or 40 ng/ml conditions as well cells that looked similar to cells in 10 ng/ml TGFβ1 or untreated (Fig. 6F). This was best exemplified at a higher magnification (Fig. 6L).
Integrin expression in slices treated with both TGFβ1 and ethanol was similar as that for nCAM. Co-treatment significantly (P < 0.05) increased the expression of integrin αv and integrin β1 at t6 as compared with untreated conditions (Figs 8 and 9). In contrast, the expression of integrin α3 and integrin β1 was significantly (P < 0.05) lower in the co-treated slices than in slices treated with ethanol at t6 and t12 and for integrin αv at t12.
Cell Migration in Controls
Desynchrony of Cortical Development
A challenge to studying cell migration in a neuroepithelium in situ is the desynchrony in the developing brain. This is evident among proliferating cells that are distributed through all phases of the cell cycle. Thus, an acute (i.e. 1 h) pulse of BrdU indiscriminately labels a subpopulation of cells that is in or recently exited from the S-phase. Further desynchrony results from post-mitotic activity. After BrdU-labeled cells mandatorily pass through the G2- and M-phases, some cells permanently exit the cell cycle and others re-enter it (Miller and Nowakowski, 1988; Nowakowski et al., 1989; Takahashi et al., 1995; Miller, 1999). This phenomenon cannot be addressed with the present model.
The movement of post-mitotic 'pioneer' cells labeled by a BrdU pulse can be traced by focusing on the first cohort of cells to leave the proliferative zones. The present study measured the change in the position of such cells between t6 and t12. By t6, migration has commenced. This is noted by the change in the distribution of BrdU-positive cells between t1 and t6, the increased presence of labeled cells in the IZ at t6, and the significant change in the mean distance of the outermost cells from the ventricular surface between t6 and t12. Therefore, by examining the change in the position of leading cells between t6 and t12, insight into cell migration can be gleaned.
Pathways of Cell Migration
An assumption in the present study is that cortical migration occurs via a radial pathway. This is supported by many studies showing that the majority of neurons in the adult cortex are generated in the VZ and SZ and migrate along radial glia (Rakic, 1971, 1972). Recent data show that many GABAergic local circuit neurons (LCNs) are generated in the ganglionic eminence and migrate via distinct routes into the immature cortex, notably by tangential paths through the IZ, SZ, and MZ (Anderson et al., 1997; Wichterle et al., 2001; Jimenez et al., 2002; Ang et al., 2003). The pathway through the IZ raises the possibility that some of the BrdU positive cells measured in this study are migrating tangentially rather than radially. BrdU labeling studies, however, show that most LCNs migrating through the IZ (where the outermost 10 BrdU-positive cells are located at t6 and t12) are generated on G14 (Tamamaki et al., 1997). This is four days before the slices in this study were exposed to BrdU. Thus, it is unlikely that LCNs migrating tangentially from the GE through the IZ are included in the present analysis.
LCNs migrating through the SZ (Anderson et al., 2001) or MZ (Lavdas et al., 1999; Zecevic and Rakic, 2001) are generated within the timeframe of BrdU exposure used in the present study (Ang et al., 2003). Hence, the contribution from these cells must be taken into account. LCNs comprise ∼10% of the neurons in mature rat cortex (Peters et al., 1985). Thus, at most, LCNs account for only one of the outermost 10 cells used in the calculation of the rate of migration. This estimate must be further reduced because (i) many LCN's are generated in the VZ and SZ and (ii) LCNs derived from the GE migrate via multiple pathways, two of which have no relevance to the present analysis, i.e. via the IZ (see above) and the MZ. Though we cannot completely discount alternative origins and migratory routes of the BrdU-positive cells analyzed in this study, the timing of the BrdU exposure and the relative contribution of future LCNs to the total migrating population reduces the impact of this confounding factor.
Though migration occurs in a discontinuous manner (Edmondson and Hatten, 1987), the net movement can be determined by examining the change in position over an extended period. In the present investigation, samples were garnered on G17 and the BrdU pulse and chase studies occurred on the equivalent of G18. In such untreated cultures, the mean net movement over six h was at a rate of 144 ± 4 µm/day. This is remarkably similar to the rate of migration in vivo for cells born on in control rats on G17. Such migrating cells move from the proliferative zones in vivo at a mean rate of 137 ± 15 µm/day (Miller, 1993, 1999).
Cell Adhesion Proteins
As with non-neural systems, cell migration during cortical development partly relies upon appropriate CAP expression (Anton et al., 1996; Zheng et al., 1996; Georges-Labouesse et al., 1998; Anton et al., 1999; Graus-Porta et al., 2001). CAPs mediate attachments formed at the leading edges of motile cells and lost at the trailing process (Lauffenburger and Horwitz, 1996). Not surprisingly, the present study shows that the IZ of the developing cerebral wall is heavily populated by nCAM-positive neurons. Specifically, nCAM is expressed at the surface of these IZ cells.
One unexpected, yet interesting, change in CAP expression in the untreated cultures is an increase in CAP expression, particularly nCAM and integrin α3, between t6 and t12. The mechanism underlying this change is unknown. Possibly, it results from the continued generation of cells that can express CAPs. Alternatively, the increased CAP expression may result from growth factors released by the slices into the medium. Serum-free medium conditioned by slices from neonates exhibits increased nerve growth factor (NGF) and basic fibroblast growth factor content (unpublished observations). It is also possible that the change in adhesion protein expression is a reflection of alterations in non-migrating cells (e.g. post-migratory neurons in the CP) or indicates changes in cellular CAP expression that do not translate to changes in cell-surface expression.
Effect of TGFβ1 on Cell Migration
TGFβ1 alters cell migration and these effects are concentration-dependent in a biphasic manner. At low and moderate concentrations, TGFβ1 promotes the rate of cell movement, whereas at high concentrations, TGFβ1 retards cell migration. Indeed, a similar concentration-dependent effect of TGFβ1 has been described for T-cell chemotaxis; exposure to low concentrations of TGFβ1 promotes chemotaxis and supraoptimal concentrations of TGFβ1 are inhibitory (Franitza et al., 2002).
Not only does exogenous TGFβ1 increase the rate of migration, it increases nCAM and integrin expression. Such an effect parallels the robust TGFβ1-mediated up-regulation of nCAM expression in simpler neural-based systems, e.g. primary cultures of developing cortical neurons (Miller and Luo, 2002) and dissociated cultures of B104 neuroblastoma cells (Luo and Miller, 1999a). Furthermore, these changes echo the effects of another member of the TGFβ family, bone morphogenic proteins, on nCAM and L1 (Perides et al., 1994).
As with cell migration, TGFβ1 effects nCAM and integrin expression in a concentration-dependent manner. Interestingly, the effects of concentration are different for cell migration and CAP expression. Whereas the effect of TGFβ1 on cell migration is biphasic, the effect on CAP expression is monophasic. That is, treatment with low or moderate concentrations of TGFβ1 induces a graded increase in both the amount of CAP expression and the rate of cell migration. At a high concentration of TGFβ1, however, the effects are divergent; the rate of cell migration falls while CAP expression continues to increase. One explanation is that there is an optimal 'working' concentration of TGFβ1 (∼10 ng/ml) to promote migration. At supra-optimal concentrations (∼40 ng/ml), migrating cells express such large amounts of adhesion proteins that adhesions are too strong to be conducive to cell motility. This conclusion is supported by a study examining the relationship between the rate of cell migration in Chinese hamster ovary (CHO) cells and the expression of integrins (Palecek et al., 1997). Increases in integrin expression only promote the migration of CHO cells to a certain amount after which higher integrin expression hinders cell migration.
Effect of Ethanol on Cell Migration
Cells in ethanol-treated slices migrate at half the rate (84 ± 14 µm/day) of cells in untreated cultures. In vivo, cells generated on G17 in an ethanol-treated rat migrate at a mean rate of 45 ± 5 µm/day (Miller, 1993). This difference between the two rates is statistically significant (P < 0.05). Interestingly, this means that the concentration of ethanol used in vitro (400 mg/dl) is less effective in slowing cell migration than is a lower exposure (∼150 mg/dl) in vivo. Thus, the present data support the use of the slice cultures as a conservative model of cell migration and fetal ethanol-induced changes. In previous studies of developing cells, it was shown that higher concentrations were needed in vitro to cause similar changes in vivo. Treatment of cells with 400 mg/dl of ethanol in vitro (Luo and Miller, 1997; Jacobs and Miller, 2001) has a similar effect on cell cycle kinetics as 150 mg/dl ethanol in vivo (Miller and Nowakowski, 1991).
One potential problem with examining the effects of ethanol on cell migration is that ethanol can suppress a contiguous developmental process, cell proliferation (Miller, 1989; Miller and Nowakowski, 1991; Miller and Kuhn, 1995). Conceivably, the change in cell migration results from an ethanol-induced decrease (i) in cell proliferation; (ii) in the number of cells that exit from the cycling population; and (iii) and in the timing of migration out of the proliferative zones (Miller, 1986; Miller and Nowakowski, 1991; Miller, 1993; Jacobs and Miller, 2001). Cell migration, however, begins in slices exposed to ethanol by t6. This is indicated by a significant change in the position of the outermost cells between t1 and t6 in the ethanol-treated slices. Thus, the ethanol-induced deficit in cell migration most likely results from ethanol-dependent changes in cell movement. Interestingly, there is a difference in the position of the outermost cells in the untreated and ethanol-treated conditions at t6. The outermost cells in the ethanol-treated condition are closer to the ventricular surface than controls. This suggests a delay in the onset of migration. Such a delay has been described as an effect of ethanol exposure in vivo (Miller, 1993).
Like TGFβ1, ethanol increased the expression of nCAM and integrin proteins in the immature cerebral wall. Note that ethanol reduces cell migration and increases CAP expression in a pattern reminiscent of that caused by high concentration (40 ng/ml) of TGFβ1. In the context of the migration model described above, one way ethanol may affect cell migration in the slice cultures is by increasing CAP expression beyond optimal amounts for cell migration.
CAPs are a target of ethanol toxicity. Ethanol affects adhesion protein expression and function in various neural systems. The most thoroughly studied CAP is nCAM (Luo and Miller, 1999a; Miñana et al., 2000; Miller and Luo, 2002). Gestational ethanol exposure alters normal expression of nCAM in the postnatal rat brain, including a decrease in expression overall as well as a decrease in nCAM cell surface expression in cultured astrocytes (Miñana et al., 2000). The functional, adhesive properties of CAMs, like L1, are also vulnerable to ethanol exposure. Ethanol disrupts L1-dependent cell adhesion without effecting the expression of the receptor, suggesting that ethanol physically blocks the homophilic interaction between L1 molecules (Charness et al., 1994; Ramanathan et al., 1996; Wilkemeyer et al., 1999).
The disruption of normal integrin expression within the developing cerebral cortex is a novel mechanism through which ethanol may interfere with cell migration. Ethanol can potentiate integrin expression by hepatocytes (Schaffert et al., 2001). The present data showing the effects of ethanol on integrin expression and cell migration are fascinating in light of recent studies of knockout mice deficient in integrin subunits. Mice lacking integrin α3 exhibit abnormal neuronal migration and premature differentiation of radial glia into astrocytes during cortical development (Anton et al., 1999). This phenotype is similar to that described in rats prenatally exposed to ethanol (Shetty and Phillips, 1992; Miller and Robertson, 1993). Likewise, integrin β1 knockout mice also feature inappropriate neuronal migration (Graus-Porta et al., 2001). Ectopic cells are common in the MZ and the pial surface is abrogated. Such migratory defects are not uncommon in human and rodent brains exposed to alcohol (Clarren et al., 1978; Kotkoskie and Norton, 1988; Komatsu et al., 2001; Miller and Mooney, 2004; Mooney et al., 2004).
Interaction of TGFβ1 and Ethanol on Cell Migration and CAP Expression
Combined treatment with TGFβ1 and ethanol results in a rate of migration similar to untreated conditions. This implies either that ethanol interferes with TGFβ1-dependent changes in cell migration or that TGFβ1 counters the depressive effects of ethanol. CAP expression is also effected by TGFβ1-ethanol co-treatment. Ethanol and TGFβ1 treatment alone increase CAP expression. Instead of an additive effect, however, co-treatment reduces CAP expression. This suggests that the restoration of a normal rate of migration in the TGFβ1-ethanol co-treated slices may be a consequence of a level of CAP expression that facilitates normal cell motility. Furthermore, the data indicated that TGFβ1 and ethanol might modulate CAP expression, and possibly cell migration, through similar mechanisms. Studies of neuroblastoma cells (Luo and Miller, 1999a,b) support this conclusion. TGFβ1 and ethanol are both anti-mitogenic and they initiate the ras/raf/mitogen-activated protein kinase (MAPK) pathway in a similar manner. Both substances induce a similar response in MAPK activity, i.e. a sustained increase.
The Endogenous TGFβ System Regulates Migration and Is the Target of Ethanol Toxicity
The developing cortex comprises TGFβ ligands and receptors (Heine et al., 1987; Flanders et al., 1991; Pelton et al., 1991; Tomoda et al., 1996; Miller, 2003). TGFβ1 is expressed by cells in the proliferative zones and TGFβIr and its sister receptor, TGFβIIr, are expressed by migrating neurons and radial glia. Thus, the TGFβ system is endogenous to the developing cerebral cortex, and it is strategically expressed to play a role in neuronal migration. Moreover, the TGFβ system is affected by prenatal exposure to ethanol (Miller, 2003). Exogenous TGFβ1 presumably interacts with the endogenous system. Furthermore, if migrating cells in the cerebral wall respond to exogenous TGFβ1 with a change in the rate of migration and CAP expression, it is probable that endogenous TGFβ1 has a similar function in vivo.
There are striking similarities between the phenotypes of the TGFβ knockouts and animals and humans exposed to ethanol in utero. Cardiac and skeletal dysgenesis as well as craniofacial abnormalities are hallmark phenotypes in the TGFβ knockout mice (Proetzel et al., 1995; Sanford et al., 1997). These defects echo malformations described in animals prenatally exposed to ethanol (Sulik et al., 1981; Sulik, 1984; Daft et al., 1986; Edwards and Dow-Edwards, 1991; Kotch et al., 1992; Astley et al., 1999) and children with FAS and ARND (Jones and Smith, 1973; Jones et al., 1973; Jones et al., 1974; Abel, 1981). Hence, the endogenous TGFβ system appears to be a specific target of ethanol.
We thank Renee Mezza for assistance with immunohistochemical techniques, Dr. Sandra Mooney for help with densitometry analysis, and Brad Pawlikowski for constructive comments on the manuscript. This research was supported by National Institute of Alcohol Abuse and Alcoholism (AA06916 and AA07568) and the Department of Veterans Affairs.


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Listen to U2 Radio on iHeartRadio
Bono's cycling accident is apparently more serious than first confirmed.
Rolling Stone reports the U2 frontman underwent a five-hour surgery for multiple breaks in his shoulder blade and upper arm. Doctors in Manhattan say metal plates and 18 screws were used in some repairs.
The rockstar also suffered a facial fracture around one of his eyes. He'll need intensive therapy, but a full recovery is still expected.
Doctors called it a "high energy bicycle accident" involving another rider in New York's Central Park.
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Me gustaría que fuera en un restaurante, o quizás en alguna Costa pues adoro el mar...
I love shopping and fine dining. Food makes me happy as does shopping!
First date something adventurous that I've never experienced before
Possibly dinner (somewhere kid friendly) or a walk in the park to get to know each other better
Very opened minded...would love to go shopping........hope u enjoy that too
We go shopping you tell me about your self n we go eating
Go to the caffee and talk, i lik fun 🏂🎿🏊🏿
go out,have fun,go watch movies,shopping and make it worth d while
It dont matter as long as its fun im up for it
I would go to a hight class fancy place, or a long drive while having a caffee and talking, to a movies etc...
Somewhere in public first date maybe coffee shop
To show me the world in one day
To have fun and to share love together.. With hanging out and to play with other
Maybe go to eat or walk around the mall get some drinks I'm always open to ideas just ask
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Kopari Colony E Thane, Mumbai
All students who cleared IPCC exam and served 18 months articleship can attend for Final exam of Chartered Accountancy Course
Passing IPCC exam is pre-requisite for attending CA final exam of Chartered Accountancy Course
Books (modules) given by ICAI, books of eminent author on various subjects. Your patients, confidence and more important your valuable time is essential for successfully passing this exam"
Student will be able to appear to exam with
proper preparation and confidence. Further,
they will get the expert knowledge of the subjects and this will help them to start jobs in the field of Accountancy, taxation, law and managements etc at higher levels
Not decided yet
CA, CS and B.com
Students Interested 1


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Big Market Research adds a new study " Global Dry Mortar Market Research Report 2016″ Order This Report by calling BigMarketResearch.com at +1-971-202-1575.
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whale tail
n.— «Garv is working on Whale Tail it's a comedy about the fashion of exposed thongs.» —by Garv I Am The Sign Stealer In Everyone Jan. 3, 2004. (source: Double-Tongued Dictionary)
whale tail
- Questions As Veiled Criticisms 02/20/2017: A psychotherapist in Burlington, Vermont, observes that couples in counseling together ask each other questions that are actually veiled criticisms. Such indirect communication was the... [more]
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|Property type
|Minimum Stay
|7 nights
Sipping cocktails by the pool while you watch the beautiful colors in the sky of the sunset over the lake will be just one of the highlights of your stay. This lovely home is decorated from top to bottom to make you feel like you are a million miles away from reality, and completely on vacation! An open floor plan makes your vacation flow with ease as you can be preparing an after beach snack, while tuning in on the card game going on on the back patio! Don't miss a beat with cable TV, wifi internet, and all of the modern needs of the modern family! With three spacious bedrooms and two large bathrooms, there is more than enough space for all guests to spread out, get comfortable, and simply enjoy peace and quiet in serine environment! Enjoy the pool with a view of the lake, surrounded by some of Florida's most beautiful landscapes and natural scenery. Take a nap or read a book, but make sure to pour that glass of wine and make it back in time for a spectacular sunset! Your vacation 'home away from home' is situated in Heron's Watch, a pristine neighborhood in a much desired area of Bradenton, Florida. This is the perfect location for an active or relaxing vacation. Only 10 minutes from the area's award winning beaches.
This is a comfortable house with a nice pool and lanai and a nice view of the pond with lots of birds, etc. Walk-in shower and the open floor plan were great. The area is quiet, but conveniently located for getting to the beach or shopping or other attractions.. The property manager was great to work with to handle all little problems.
It was great and we felt very good.
A really great home to stay for vacation. Wonderful spacious bed rooms to retire and relax, a great and comfortable living room and very good equipped kitchen. The pool and terrace invite for relaxing hours after staying at the beautifull beaches of Ana Maria Island, which are only a few minutes away. Really everything is available within a few minutes ride, restaurants, pubs, groceries, beaches.
We really loved this place and it was really hard to leave back home to Germany.
We spent 2 months in this lovely well stocked home. The lanai with nice furniture and pool and great view of the pond with birds and turtles and fish, the comfy open floor plan, the spacious master bath, the quiet neighborhood only a few miles from the beach, the attachedd garage, and the very helpful property manager all contributed to a super stay. It was indeed our home away from home. We have already reserved it for a month next winter.
Whoever decorated sure has a talent! So much to see in this home! Very pretty and the property was well maintained. Everything was as expected. Right up Cortez road (and 75th street) there was a shopping center we were at almost every day. It has a grocery store, MDonalds, Starbucks, Bonefish Grill, Marshalls, nail salon, hair salon, and liquor store! It was awesome!
Beautiful home in a quiet little neighborhood close to the beach. Couldn't think of a more ideal home and location for the price. Everything in the home was very thoughtful from the welcome package to how clean it was. We really liked First Watch for breakfast. It's just up Cortez Road in the shopping plaza with McDonalds and Starbucks. (yes, that was nice too!)
Year-round sunshine, surf, and leisure sports lure visitors to Bradenton, a friendly tropical spot south of Tampa Bay on Florida's Gulf Coast. Hernando DeSoto discovered the area in 1539, an event commemorated each spring with the month long Florida Heritage Festival. Stroll along the waterfront downtown, take up canoeing, fish for grouper and mahi mahi, play golf or just relax and watch the sun dip into the sea. Kids of all ages will enjoy visiting Snooty, the South Florida Museum's star manatee.
|Rental Period
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|Additional Nights
|Weekend Night
|Monthly *
|Event Price
High Season
1 Feb 2018 - 20 Apr 2018
7 night minimum stay
for 7 nights
My Standard Rate
7 night minimum stay
for 7 nights
|Damage Waiver
|£40 + Tax
|Professional Cleaning
|£121 + Tax
|Booking Fee
|£39 + Tax
|Tax Rate
* Approximate monthly price. Actual pricing will depend on the days of the month you stay.


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All properties may be subject to local taxes, fees and a $9.99 utility processing fee. This is a bright haven with adorable window shutters on the front of this single-story home. Open the door and feel the warm welcome that emanates from the carpeted great room in this open-concept design that allows natural light from broad windows to work at maximum strength. The dining area has a delicate chandelier and is well-lit by the the long window harbored in the back door which opens up to the spacious backyard. Black appliances, a broad breakfast bar, and ample cabinetry make this a convenient kitchen that is ready to fulfill all your needs. A generous walk-in closet can be found in the master suite where you will also find an attached bathroom. For a cozy, cheerfully bright living space, check out this home on Oak Hill Lane! For other quality homes for rent, visit www.americanhomes4rent.com. Equal Housing Opportunity.
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OBJECTIVE To pilot, among women with gestational diabetes mellitus (GDM), the feasibility of a prenatal/postpartum intervention to modify diet and physical activity similar to the Diabetes Prevention Program. The intervention was delivered by telephone, and support for breastfeeding was addressed. RESEARCH DESIGN AND METHODS The goal was to help women… (More)
OBJECTIVE To examine the association between breastfeeding intensity in relation to maternal blood glucose and insulin and glucose intolerance based on the postpartum 2-h 75-g oral glucose tolerance test (OGTT) results at 6-9 weeks after a pregnancy with gestational diabetes mellitus (GDM). RESEARCH DESIGN AND METHODS We selected 522 participants enrolled… (More)
BACKGROUND Women with gestational diabetes (GDM) are at high risk of developing diabetes later in life. After a GDM diagnosis, women receive prenatal care to control their blood glucose levels via diet, physical activity and medications. Continuing such lifestyle skills into early motherhood may reduce the risk of diabetes in this high risk population. In… (More)
OBJECTIVE To examine the risk and etiology of preterm delivery in women with polycystic ovary syndrome (PCOS). STUDY DESIGN Retrospective cohort study comparing preterm delivery rate among non-diabetic PCOS and non-PCOS women with singleton pregnancy. Multivariable logistic regression was used to identify predictors of preterm delivery among PCOS women.… (More)
OBJECTIVE To compare the effectiveness of diabetes prevention strategies addressing postpartum weight retention for women with gestational diabetes mellitus (GDM) delivered at the health system level: mailed recommendations (usual care) versus usual care plus a Diabetes Prevention Program (DPP)-derived lifestyle intervention. RESEARCH DESIGN AND METHODS… (More)
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Seattle FHLBank Hires Black Rock, OCC Veteran
The Federal Home Loan Bank of Seattle has hired Black Rock Financial and a firm headed by former Comptroller of the Currency Eugene Ludwig to assist it in getting its finances and management practices in order.
According to officials familiar with the situation, Black Rock, including its managing director, Peter Fisher, is helping the GSE with its three-year business and capital management plan.
"Peter has been very helpful" to the FHLB, said one source.
Mr. Fisher is a former undersecretary at the Treasury Department and was also a top official at the Federal Reserve Bank of New York. (Black Rock's clients include Freddie Mac.)
Promontory Financial Group of Washington, Mr. Ludwig's group, is working on a review of the Seattle bank's management.
The two firms, however, are not involved in an investigation of FHLB stock sales by three depositories that may've redeemed stock based on nonpublic information they had access to.
That review is being conducted by five nonmember directors with the assistance of an outside law firm.
(c) 2005 Mortgage Servicing News and SourceMedia, Inc. All Rights Reserved. http://www.mortgageservicingnews.com http://www.sourcemedia.com
Decisions, Default Rates, & Debt-To-Income The QM Rule's Impact


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Jean-Pascal Imsand, Photographer
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Jean-Pascal Imsand, Photographer by Fondation Jean-Pascal Imsand
Book DescriptionAmong those Swiss photographers who came to the fore in the 1990s, Jean-Pascal Imsand was always something of a loner--both professionally and in his life. Born in Lausanne, he lived and worked mainly in Zurich and on the Lake of Geneva. Imsand's journeys abroad were few; the world in which he moved with ease was one of local neighbourhoods. He travelled to Istanbul, Venice, Paris, and New York for purposes of private research and commissions for portraits. Jean-Pascal Imsand's work eludes any attempt at classification, and contains an apparent paradox. While his photo-montages are much admired for their aesthetic and narrative qualities, they are often political in nature; the precision of content demonstrated in his socio-documentary reportages does not exclude mythical interpretation. These self-assigned works mark the end of the artist's creative period, which lasted a mere decade. Having ended his own young life, Jean-Pascal Imsand's oeuvre remains a fragment, but the fragment was one of his working principles. The principal sponsor of this catalogue is the Banca del Gottardo.
Buy Jean-Pascal Imsand, Photographer book by Fondation Jean-Pascal Imsand from Australia's Online Bookstore, Boomerang Books.
Book DetailsISBN: 9783037780374
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Publisher: Lars Muller Publishers
Publish Date: 1-Jun-2004
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text: Tech Yahoo's Mayer Steadies Ship By Amir Efrati And John Letzing Updated Jan. 29, 2013 4:09 a.m. ET Yahoo Inc. Chief Executive Marissa Mayer appears to be holding the ship together. Six months into Ms. Mayer's tenure at the helm of the Internet company, Yahoo posted an 8% profit drop and a 2% revenue increase for its fourth quarter. ... RELATED VIDEO Yahoo Tops Analysts' Estimates Yahoo Inc., under the leadership of CEO Marissa Mayer, posted quarterly results that topped analysts' estimates. WSJ's Amir Efrati reports. (Photo: Getty Images) To Read the Full Story, Subscribe or Sign In Most Popular Videos The Moment Kim Jong Nam Was Attacked: CCTV Footage Light Plane Crashes Into Shopping Mall in Australia French Cafe Receives Michelin Star by Mistake Anti-Trump Message Projected on U.K. Parliament Building A Child Is Pulled Alive From the Rubble in Syria Most Popular Articles Nascar, Once a Cultural Icon, Hits the Skids Trump Administration Tightens Deportation, Detention Rules Kraft-Unilever Deal Is Off, but Warren Buffett's Anomalies Live On Opinion: Stop Chuck Todd Before He Tweets Again Trump Lashes Out at Sweden Over Immigration


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First A320 assembled in China performs maiden flight
First fuselage section for the A330-200F freighter is delivered
First fuselage section for the A330-200F freighter is delivered
"QUOVADIS" - the new RNP services company by Airbus
Developing RNP activity and procedures for authorities, airlines and airports From July 2009, QUOVADIS, a new 100 percent subsidiary of Airbus, based in Toulouse, will sell and provide 'Required Navigation Performance' (RNP) services to authorities, airlines and airports, ranging from RNP ...
New payload/range capability for the A330-200
New payload/range capability for the A330-200
Five teams reach Airbus' "Fly Your Ideas" contest final
Teams hail from the Asia-Pacific, North America and Europe Five teams out of the 86 that were still in the competition have been chosen to go forward to the grand final of the Fly Your Ideas challenge at the Paris Le Bourget Airshow. Airbus, launched the contest in October 2008. More than 2,350 stud...
Airbus changes A380 delivery schedule for 2009/2010
Due to the current economic and aviation crisis and following specific customer requests for deferrals, Airbus is adapting its A380 aircraft delivery schedule for 2009/2010. According to the new plan, Airbus expects to deliver 14 double-decker aircraft in 2009 and more than 20 aircraft in 2010. Onwa...
"Fly your ideas" university contest ready for final round
Ten expert-jury from around the world to evaluate students' eco-efficient ideas Airbus, one of the world's leading aircraft design and manufacturing companies, is pleased to announce the jury members for the final of its international student competition, "Airbus Fly Your Ideas." The compe...
Airbus/EADS sign a Titanium supply agreement with VSMPO-AVISMA, integrated structure of the Russian Technologies State Corporation
Joint press release issued by Airbus, EADS, VSMPO-AVISMA and Russian Technologies State Corporation
Swiss takes delivery of its first A330
A 'first class' aircraft for a 'first class' airline Zurich based Swiss International Air Lines has received its first new A330-300. The aircraft is the first of nine A330-300s ordered in September 2007 to expand the Swiss' medium to long haul network. Powered by Rolls-Royce...
The A330/A340 Family jetliners benefit from lower maintenance costs
Optimisation of scheduled maintenance programme intervals for A330/A340 Family jetliners has resulted in improvements in maintenance costs for these Airbus aircraft, which also contributes to their efficiency.


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XCOM: Enemy Unknown
What parents need to know
Parents Need to Know
Parents need to know that XCOM: Enemy Unknown is a mobile version of the popular console and PC game. It was an M-rated game then, and nothing has changed. The focus is on strategy, but there's strong violence, gore, and blood, and deaths are violent. Strong language is easily found. There's no multiplayer component, but this is still much too mature for young players.
User reviews
Parents say
Kids say
What's it about?
You command troops on the ground, pointing at the spot on the map you'd like individual troops to move to, and then you choose from a menu what your next action will be. Since the game is turn-based, there's no rush to decide, and many options can be considered. In non-combat situations, players oversee the research of alien artifacts and the manufacturing of products to better protect and arm the troops; you also monitor for activity.
Is it any good?
When it hit PC and console in 2012, XCOM: ENEMY UNKNOWN topped many "game of the year" lists, and this pitch-perfect mobile version incorporates all that the previous version got right. With an incredible blend of action and strategy, it's a game that requires players to think carefully as they fight little green men on the battlefield and research and build new weapons and satellites at their operational headquarters. Heck, they even need to give thought to the best troops to send into the field.
The graphics take a bit of a hit from the earlier version but are still fairly impressive for tablets. Also, though the high cost might scare some people, this is a game that's easily worth the $20 price tag. Still, it's very violent and definitely too much for kids.
App details
|iPhone, iPod Touch, iPad
|Release date:
|June 20, 2013
|Strategy Games
|2K Games
|Minimum software requirements:
|iOS 5.0 or later
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Rated 'Excellent' on Trust Pilot
"Great service, great organic vegan raw ingredients. Delivered within a few days. Highly recommend this company."
Susan Jane Duckworth,Published 20 October 2016
Pineapple and Lime Shampoo is a wonderfully uplifting blend of organic Pineapple, Lime and Mango. It's like sunshine in a (recycled) bottle. No artificial colours or fragrances.
Pineapple & Lime Shampoo Vegan ingredients: Ammonium laureth sulfate, Ananas sativus fruit extract, Aqua, Citric acid, Citrus aurantifolia oil, Cocamidopropyl betaine, Curcumin, Glycerin, Glyceryl laurate, Helianthus annuus seed oil, Limonene, Linalool, Mangifera indica fruit extract, Maris sal, Parfum, Polysorbate 20, Potassium Sorbate, Sodium benzoate, Tocopherol
In Stock


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Mobile Welfare Pod (Diesel)
The Elliott Evolution Mobile Welfare Pod, provides the comfort of a ceramic flushing toilet and convenience of a changing room in this 6 person, self powered mobile welfare unit.
This mobile welfare facility can easily be towed allowing instant welfare facilities to be provided for workers moving around on a large site. The Anti-Vandal design and security features are enhanced with its safe and secure kneel down axle system and retractable drawbar.
Our Evolution Mobile Welfare Pod includes:
- Water tank 80 ltr
- Effluent tank 100 ltr
- Fuel tank 25 ltr diesel
- Generator 5.9 KVA
- Generator powered (included)
- HSE compliant
- Anti-Vandal 12'6" x 7' towable
- Canteen area for 6 people
- Ceramic flushing toilet
- Changing room
- Silenced generator
- Integral water tank
- Integral effluent tank
A range of portable cabin accessories and value added products are also available. Click here for more information.
To read more about our most popular Self Powered Units (including layouts, sizes and weights) please refer to pages 50 - 52 of our Hire Product Guide.
Click here to download.
Evolution welfare units can be ordered from your local Elliott portable cabins branch or through our specialist portable toilets and showers team. Please visit our Contact Us page for contact details.
I want to echo what has been said about our Elliott classroom. The installation team were superb and everything went smoothlyHazel Scott - Ringway Primary School
Thank you for your incredible efforts and hard work at The 2014 Ryder Cup. You have gone above and beyond what we expected and cannot thank you...Chris Atkinson, The Ryder Cup 2014
Elliott were professional and extremely helpful, making Race for Life exceptionally smooth. We wouldn't hesitate to recommend them.Ruth Chamings - Cancer Research UK
You and your team have worked tirelessly to erect and fit out the temporary classrooms in the very limited timescale that we set. Over the last...John Wright
Thank you to Elliott for providing the portable toilets for the Woolfest event at Woolton Hill Junior School which was a huge success. We look...Jo Godfrey
A big thank you to all your staff who kept the loos clean, filled up and available all through Car Fest South this weekend and to your wonderful...Kathy Worrall
On behalf of all at Vision Nine HQ, I'd like to thank you and the team for your services and efforts that made Boardmasters 2013 such a great...Joe Baran
Thank you to the whole team involved in installing our two modular classrooms, both staff and students are very appreciative of the modern,...Barbara M Belfield-Dean, Concord College
Due to business expansion within the two sites in Chatteris, Cambridgeshire, Erms UK Ltd needed to engage with robust and reliable partners. For...Phil Barton Carroll
I must comment on the superb service from your staff. All of which dealt with my order very efficiently. I have no hesitation in recomending...Neville DeLord, St Columbas College
The driver who delivered the cabin was extremely polite and professional and nothing was too much trouble.Rachael Sheppard, MOSI
Excellent service from your office staff and delivery driver. A good clean and smart looking unit that is practical and meets my requirements....Andy Mitchell, Hill Partnerships Ltd
I am writing to express my gratitude to your company for the professional way they handled our event. This was a high profile event, particularly...Sue Hayden
"Elliott's have worked with Mace in-conjunction with Hertfordshire County Council on a 7-year £30m framework which involved over 30 projects...Pino Bonsignore, Project Lead Designer, Mace
"McDonalds selects it suppliers very carefully and we chose Elliott because of its proven ability to manufacture high quality volumetric units to...Richard England, New Build Construction Manager – McDonalds UK
"Elliott Off-Site Building Solutions delivered a high quality three-storey modular office building, in a rapid timescale, on time, on budget and...Ralph Smith, Project Manager – Jaguar Land Rover
"Prior to the delivery, I was very impressed with the friendly helpful person/people I spoke to on the phone. Everything was on time, efficient...Anita Horsnell
"European Tour Productions were more than satisfied with the service we received from Elliott Hire at the Ryder Cup. The cabins and furniture...Tony Coxon, European Tour Productions
"From my first contact with people at Elliott I found they were extremely helpful and gave all the information that I required to hire a portable...John Sterry


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Jan's Storytelling Club
By Unknown
To listen to an audio podcast, mouse over the title and click Play. Open iTunes to download and subscribe to podcasts.
Your Story Listening Space
|Maria's Gift
|Maria's Gift Ages 7 and Up Maria's life is hard but she faces it with courage. She does all that she can for her family, despite the cruelty of others, there in that little village where she lives. Then she goes a journey and meets unusual strangers. What will that meeting bring her? Click on the […]
|2 5 2012
|View in iTunes
|Nagila and Navoma and Na
|Nagila and Navoma and Na Ages 9 and up Three young women make wishes for when they are married. Their wishes are granted. They even get to marry a prince. But will this bring them happiness? Click on the link and find out. A story from Africa. 11:42 mins. This story has to do with […]
|16 4 2012
|View in iTunes
|Gold Tree And Silver Tree
|Gold Tree and Silver Tree Ages 11 and up A queen and her daughter go walking. They come to a glen and a brook. "Am I not the fairest in all the world?" the queen asks the fish that swims there. "No," says the fish, "You are not." What happens next? Who is the fairest? […]
|14 3 2012
|View in iTunes
|Lion, Hyena and Rabbit. A Story In A Story
|Lion, Hyena and Rabbit. A Story in a Story Ages 11 and up Three animals decide to hunt together but then there is a problem. This is a story that is more serious than you might think when you first hear it. It's a story that meant something special to at least one listener. Click […]
|6 3 2012
|View in iTunes
|The Salmon Cariad
|The Salmon Cariad Age 7 and up On a moonlit night, a young man goes to the river to fish. He catches a salmon but what happens next is beyond his imagining. He goes a journey he was not expecting. Where does he travel? Click on the link to listen and find out. A story […]
|7 1 2012
|View in iTunes
|Tiddy Mun
|Tiddy Mun Ages 7 and Up Scary creatures live in the marshes but Tiddy Mun is the people's helper. He walks at night, they hear him laughing. He has saved them over and over again but will he come to save them now? Click on the link to listen and find out. A story from […]
|25 11 2011
|View in iTunes
|Wise Father, Wise Daughter
|Wise Father, Wise Daughter Ages 7 and Up Paul comes to Canada because he has been driven out of his own country for the colour of his skin. He starts a business but he is growing older. How will he choose which one of his daughters should inherit? Click on the link and listen to […]
|9 11 2011
|View in iTunes
|The Old Woman In The Woods
|The Old Woman in the Woods Ages 7 and Up When Jack gets turned out of home for his laziness he meets an old woman in the woods. First she asks him to take off his jacket, then it's his boots and then…. What comes next? You'll have to click on the link and […]
|28 10 2011
|View in iTunes
|How Ti-Jean Became A Fiddler
|How Ti-Jean Became a Fiddler Ages 7 and up Ti-Jean's brothers set out to seek their fortunes but they are more likely to be thrown in prison than win the promised bride. Can T-Jean help? He certainly means to. Where does the fiddle come in? Click on the link to find out. A story from […]
|18 10 2011
|View in iTunes
|The Legacy
|The Legacy Age 9 and up The advice of a dying old man to his son seems simple but there is a riddle in it that holds a richer gift. A story from Palestine. Click on the link and listen. 3.41 mins. Was the advice wise? Would you follow it? Leave a comment for […]
|4 10 2011
|View in iTunes
|Ivan Crookback
|Ivan Crookback Age 7 and up Ivan goes looking for a way for a way to change himself. The journey is long but he is determined. What will he find? Will he be rewarded? A new twist on an old story from Brittany. 13.26 mins. I reshaped this story because I have friends like Ivan, […]
|14 9 2011
|View in iTunes
|The Young Horseman
|The Young Horseman Age 7 and up The Young Horseman sets out from his village, seeking for a wife. He meets an old man on the road. Will the old man help him? What will happen when he picks those lemons from that tree? Click on the link to listen. An African story. 10.10 […]
|6 9 2011
|View in iTunes
|Horse For Sale
|Horse for Sale Age 7 and up. A terrible plague has struck this boy's village. Everyone has been blinded. He must go on a journey to help them but he must remember his grandfather's instructions every step of the way. Can he succeed? What will happen? Click on the link and find out. A story […]
|24 8 2011
|View in iTunes
|Jack and the Walnut
|01 Jack and the Walnut Age 9 and up As always, Jack gets himself into trouble. He wants to help his elderly mother. He causes havoc instead. Recorded live. A story from Scotland. 6.55 mins Did this story make you think? Leave a comment below. Story origin for me: Scottish teller Bea Ferguson who […]
|22 6 2011
|View in iTunes
|The Child Who Looked to Herself
|01 The Child Who Looked to Herself 2 Age 7 and up. Her family is poor and she is the youngest of five sisters. Winter is upon them and she has so little to wear. What will she do? How will she manage? Click on the link and find out. A story from Greece. 8.10 […]
|22 6 2011
|View in iTunes
|The Magic Wand
|The Magic Wand 2 Age 9 and up. A magician comes to a market and he has a magic wand. A homeless boy comes to watch him. Can the magician help? Click the link, listen and hear. A story from Africa. 2.20 mins Why does that boy want what he wants? Leave a […]
|22 6 2011
|View in iTunes
|Cap O'Rushes
|Cap O'Rushes 3 Age 7 and up A daughter is left to wander the world when she makes her father angry. She comes to a marsh where reeds are growing. What will happen? Can she prove her father wrong? Click on the link to find out. A story from England. 11.53 mins. This […]
|22 6 2011
|View in iTunes
|The Old Woman and the Rock
|The Old Woman and the Rock Age 7 and up World Maker has left the old woman alone on her island. She asks for a rock as her companion. "You can have any rock," World Maker tells her but then he wants to change his mind. Click on the link to find out what […]
|22 6 2011
|View in iTunes
|The Selkie Wife
|01 The Selkie Wife Age 11 and up. She is a seal and her skin is stolen. Now she must live upon the land. Can she forget, be happy? Click on the link and listen. A story from Scotland. 7:34 mins What do you think about the ending? Leave your comment below. Story origin for […]
|19 5 2011
|View in iTunes
|19 Items
- Free
- Category: Arts
- Language: English


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Rising Arc Marketing crafts custom marketing solutions for small businesses and startup companies.
Our solutions include Done for You services as well as tools for the Do it Yourself entrepreneur.
Brand & Identity
software & Services
Marketing Campaigns
Web Design
What We Offer
Rising Arc Marketing equips small businesses and startups with the the solutions that business owners and entreprenuers need in early stage companies.
Making a professional web presence, brand marketing and product awareness campaigns financially accessible to every business in New England.
Our team lives collaboration. From the company leadership down to the newest intern, everyone contributes to organizational development and the creative process. Our team is distributed across the United States with the majority residing in New England.
Understanding your business and its unique aspects enables us to craft the solutions your specific business needs
Our process: aim, then Fire
Providing insight to enable you to make highly informed decisions on how you apply available resources.
High quality execution and delivering improvements in your business and clear return on your marketing investments.
Our Services
Business Listing & reputation building
Create & correct listings on hundreds of sites
Generate new reviews, suppressing the negative
Full monitoring capabilities with real time update notifications
Done for you services available
Social Media Management & Lead Gen
Publishing to Major Social Networks Simultaneously
Scheduler to plan posts ahead
Unique Real-Time Lead Generation
Done for You services available
Digital Branding & Advertising
Smart Target
The newest ad-tech available.
(One to One Marketing)
Google Adwords, Bing and Social Media Advertising
Email marketing, list building, SEO/SEM, SMM, Retargeting & More
A Website for Every Business
Web Design on WordPress
(Tons of extras)
Rising Arc Platinum Web
(Fully managed design)
Membership, eCommerce & more
Multiple Payment Options
Rising Arc Marketing is the Official Marketing Partner of the
New England Anchors
This partnership enables our customers to leverage the power of sports in branding campaigns without the typical oppressive cost.
Frequently Asked Questions
We get a range of questions about our available services and products. While we have tried to capture the recurring questions here, you can always contact us with specific questions.
Actionable Do It Yourself Resources Directly to You
We send once a week emails that help you build the knowledge to do it yourself, with or without our tools.
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What kind of clients do you work with?
We select and develop products and services we believe can add value to a small business owner's efforts to expand their business. We are happy to work with any business where both sides feel our team can add value and show ROI. However, we pride ourselves and measure our success on our ability to help entreprenuers who are struggling to exceed their current plateau.
what is the difference between DIY and DFY?
These terms are to differentiate the range we have in our ability to help your business at different levels of engagement and cost structures. Your business is unique and your needs are unique. Through the evaluation and planning process we'll plan the services and solutions you actually need. We don't implement boilerplates.
DFY (Done for You) are services (aka professional services or "solutions") much like you'd engage a traditional agency to provide. These would be end to end services where we manage the function based on instruction and collaboration with you and your staff.
DIY (Do It Yourself) are services focussed on helping you implement a tool into your busienss practices and training you and/or your staff to leverage the full capabilities of it. This obviously helps curb costs, but increases the workload demand on you and your team.
It can be as little as a single meeting, conversation or conference call. If we are helping you plan and design a marketing campaign or design a new website with broad or customized features, the process of design and review iterations can be more time consuming.
Getting to action as quickly as possible is always important. The action stage is also the most time consuming. Sometimes that one more night to sleep on it, or that last collaborative review can save a project a week of redesign. We like to aim before we shoot.
Why sports marketing
Sports teams typically find themselves with an loyal fan base unlike most other businesses. This makes co-branding alongside a sports effective highly effective and very popular. The costs of these partnerships represents the unique position of sports as a marketing medium, putting the opportunity out of financial reach for most small business owners.
Our partnership with the Anchors allows our team to leverage the power of sports marketing while enabling our ability to deliver this access at a price point well within reach for any business.


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Casting has been announced for the upcoming Manhattan Theatre Club production of the dark comedy To Be Or Not To Be, coming to Broadway in September. Craig Bierko and Jan Maxwell will star in this play version of the film of the same name, with Kristine Nielsen, Steve Kazee, Peter Benson, and Rocco Sisto co-starring. Bierko has been seen on Broadway in Thou Shalt Not and The Music Man, while Jan Maxwell has most recently appeared on Broadway in Coram Boy and Chitty Chitty Bang Bang. To Be Or Not To Be, adapted for the stage by Nick Whitby, will have its official Broadway opening on October 2.
The following two tabs change content below.
Pawdesh Salawi
Broadway Theatre Writer at New York Show Tickets Inc.
Pawdesh has been writing about Broadway ever since seeing her first show on the great white way in 2001. Now she sees over one hundred Broadway (and off-Broadway) shows a year. She is also a member of the Drama Desk and would love to vote on the Tony's one day. The Office Voted Her Most Likely To: Marry rich and divorce young
Latest posts by Pawdesh Salawi (see all)
- New Musical "Come From Away" Commences - February 18, 2017
- Six New Broadway Shows to Begin in February - January 28, 2017
- "Jitney" Opens on Broadway to Rave Reviews - January 21, 2017
- Tony Winning Play "The Humans" Concludes - January 14, 2017
- "Matilda," "Something Rotten!" Close New Year's Day - January 1, 2017


text: Seller: kiri67 (1,867) 99.3%, Location: Strasbourg, Ships to: Worldwide, Item: 112216884864 Beautiful vintage lot of 2 french electric twist light switch for desk / table lamp, made of plastic. white color.Lenght 6.5 cm (2.50) weight 35 grs.Very nice item, very good condition for age (see pictures). Thanks for looking and Good purchases ! Have a nice day !!!! apps.froo.com | Froo www.froo.com | Froo Cross Sell, Free Cross Sell, Cross promote, eBay Marketing, eBay listing Apps, eBay Apps, eBay Application Condition: very good condition, Color: White, Maker: brun, Material: plastic


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$899.00 $999.00
BA 115. This powerful combo features a 150-Watt power section and a single 15" Ampeg speaker that can rock a small- to medium-sized venue. Just... View full product details
Email Us for Availability $1,449.00
AMPEG BA 210, 450 WATT COMBO, 2 X 10" Ampeg's BA-210 2x10″ Bass Combo delivers classic Ampeg tone and performance-driven features in a powerful 450W... View full product details
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BA108.The 20-watt Ampeg BA-108 bass combo is perfect for beginning bassists, offering tools like 3-band EQ, CD/MP3 input and a headphone out for silent practice.... View full product details
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BA110. The 40-watt Ampeg BA-110 bass combo is perfect for beginning bassists or as a practice amp alike.offering tools like 3-band EQ, CD/MP3 input and... View full product details
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Fantastic small set up with enough power for small rehearsals. Great tone and easy to use.ۢۢ 50 WATTS, Single 12" speaker. Beautiful Ampeg tone!!! Classic... View full product details
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Ampeg Heritage SVT-410HLF Designed and assembled in the U.S.A., the Ampeg Heritage SVT-410HLF is the ultimate in premium, full-range Ampeg tone via custom-made U.S. Eminence... View full product details
$3,179.00 $3,749.00
The real deal! Designed and assembled in the U.S.A., the Ampeg Heritage SVT-810E delivers legendary Ampeg tone in a premium package. Fully-stocked with eight custom-made... View full product details
$5,299.00 $5,999.00
Ampeg Heritage SVT-CL Designed and assembled in the U.S.A., the Ampeg Heritage SVT-CL is built with premium components and construction, delivering true Ampeg tone. Premium... View full product details
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AMPEG PF 50T ALL TUBE HEAD The legendary tone and portability of the all-tube Ampeg Portaflex amplifiers is faithfully reproduced in the PF-50T. This all-tube... View full product details
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Drawing from the classic design of Ampeg۪s legendary Portaflex cabinets, the PF-210HE combines vintage styling with the performance of a modern 450W cab. The dual... View full product details
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Drawing from the classic design of Ampeg۪s legendary Portaflex cabinets, the PF-115HE combines vintage styling with the performance of a modern 450W cab. The Eminence... View full product details
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The Ampeg PF-115LF bass enclosure delivers the performance, looks and affordability of the popular Portaflex series in a powerful, shelf-ported design that hits hard and... View full product details
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The Ampeg PF-410HLF bass enclosure delivers the performance, looks and affordability of the popular Portaflex series in a powerful, shelf-ported design that hits hard and... View full product details
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The PF-800 delivers the superior performance of Ampeg's Portaflex Series bass heads in an ultra-compact 800-watt design perfect for power-hungry bassists that demand uncompromising tone.... View full product details
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Ampeg۪s Portaflex PF-350 delivers legendary Ampeg tone in an ultra-compact design that pushes out a hefty 350W. Tools like a built-in limiter, FX loop and... View full product details
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Ampeg۪s Portaflex PF-500 bass head delivers a staggering 500W of power in an ultra-compact design providing pure Ampeg tone in an extremely portable package. Precise... View full product details
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The Ampeg PortaFlex Rig This is a great set up for everything from small gigs to big halls. The PF500 can deliver 500 watts of... View full product details
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Ampeg۪s PN-410HLF is a modern, lightweight bass enclosure built to handle the power of high-output Ampeg bass heads. Designed and assembled in the U.S.A. out... View full product details
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Ampeg SCR-DIThe Ampeg SCR-DI Bass DI with Scrambler™ Overdrive is a true Ampeg preamp, delivering a wide range of Ampeg tone perfect for the stage.... View full product details
$989.00 $1,099.00
When only the fat, warm sound of a 15 speaker will get the job done, we highly recommend the SVT-15E. Armed with a massive five-pound... View full product details
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The Ampeg SVT-212AV bass enclosure delivers pure Ampeg tone in a ported cabinet design that bridges the gap between the fast transient response you want... View full product details
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SVT 3 PRO,ۢۢ_The most popular Ampeg head that we sell. The SVT3Pro features the same tube preamp as is in the flagship SVT2Pro. It uses... View full product details
$2,999.00 $3,999.00
SVT 4 PRO. RMS Power Output: 300-Watts x 2/900 (mono-bridged @ 8 Ohms)RMS Power Output: 490-Watts x 2/1200 (mono-bridged @ 4 Ohms) RMS Power Output... View full product details
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This is the speaker cabinet for the bass player who demands the ultimate in a single, compact cabinet setup. The SVT-410HLF delivers devastating low-end bass,... View full product details
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SVT CLASSIC.ۢ The direct descendant of the 1969 model. 300 watts of all tube power, absolute devastating low end and a tone that only tubes... View full product details
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SVT 7 PRO.ۢۢ_ The SVT-7PRO pumps out a ferocious kilowatt of pure Ampeg tone in a compact design that weighs in at just 15lbs. The... View full product details
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SVT VR. This is the amp that launched a thousand careers, and it's just as good as it was way back then. The new SVT-VR... View full product details
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Looking for a speaker cabinet to brighten up your bottom-heavy bass rig? Or, maybe you۪ve recently downsized your transportation to save a few bucks on... View full product details
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For those where 410's just ain't enough and 810's is just too damn heavy. The SVT610HLF is a truely awesome cab, it creates and delivers... View full product details
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The Ampeg V-4B 100W All-Tube Bass Head is a remarkable reissue of the legendary 1971 original, delivering the pure tube tone and unmistakable grind that... View full product details


text:
Question about Creative Labs WebCam Live!
Yellow line on center of reciepients face
Posted by taylormadebc on
SOURCE: I have freestanding Series 8 dishwasher. Lately during the filling cycle water hammer is occurring. How can this be resolved
a 6ya expert can help you resolve that issue over the phone in a minute or two.
best thing about this new service is that you are never placed on hold and get to talk to real repairmen in the US.
the service is completely free and covers almost anything you can think of (from cars to computers, handyman, and even drones).
click here to download the app (for users in the US for now) and get all the help you need.
Posted on Jan 02, 2017
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In one comment Monday, the most polarizing coach in the NFL demonstrated why he is loved by some and despised by others.
"As great as today feels… we're five weeks behind the other teams for the 2017 season," the New England Patriots' Bill Belichick said, scarcely 10 hours after the Patriots pulled off the biggest comeback in Super Bowl history, miraculously erasing a 25-point deficit to defeat the Atlanta Falcons, 34-28, for a fifth Lombardi Trophy.
Whether it was sincere or simply for effect, Belichick wanted to let the world know that his warm afterglow is measured in minutes, not months. The franchise that most of the country loves to hate is already on to next...
San Diego Chargers quarterback Philip Rivers (17) hands the ball off to running back Ryan Mathews (24) during the first quarter against the Miami Dolphins at Qualcomm Stadium.


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Is Amalgamated Impex Sdn Bhd incorrect or missing key business information? Update your Hotfrog listing today.
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Is Amalgamated Impex Sdn Bhd in No. 34, Jalan PKNK 1/12, Perusahaan Ringan Ria Jaya, Sungai Petani Kedah 08000 your business? Claim your listing and attract more leads by adding more content, photos and other business details.
We have more Retail services in Sungai Petani Kedah available on Hotfrog MALAYSIA. You can update your search for Sungai Petani Retail by location, keyword or service options.
47, Lorong Gamelan 3, Taman Ria Jaya, Sungai Petani Kedah 08000
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5-E, Jalan Kampung Baru, Sungai Petani Kedah 08000
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You'll Cheer This Homeless Man While He Celebrates Getting a Job
There is nothing like the feeling when you land a new job.
There is nothing like the feeling when you land a new job.
All proceeds from the tournament went to the Hendrix family in memory of John 'Porky' Hendrix.
We know one thing for sure, there's a possessive apostrophe involved.
It was a holiday weekend so sometimes we like to get out of town and visit other places. There were a few Montanan's where I went over the weekend.
There is nothing I love more than being able to help out animals, and our friends at AniMeals in Missoula are looking for some specific items.
Jake Owen and his daughter, Pearl, have welcomed an adorable new addition to their family.
You have to have a lot of self-confidence if you're going to title your travel piece the "perfect U.S. road trip."
Lots of green attire is encouraged!
I have been thinking and I love a lot of winter things and sports per say that I like. You can snowmobile, ice wind-surf ( I guess?), ski, ice fish, winter fly fishing, curling, hockey. Which one is my favorite though?
Another year in the books for Dancing with the Missoula Stars! I was able to grab some fun photos before we started the show and a few after. Kuddos to this great community for a wonderful event.
Major League Baseball spring training is right around the corner, and in the year 2000, that meant country superstar Garth Brooks reported for duty to the New York Mets clubhouse. It was the second of three stints the singer had in the bigs, and he left with an unimpressive batting average.
How would you like to help Yellowstone National Park while you work, learn and play there all at once?