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Transplacental transfer is the fetus' primary source of taurine, an essential amino acid during fetal life. In intrauterine growth restriction (IUGR), placental transport capacity of taurine is reduced and fetal taurine levels are decreased. We characterized the protein expression of the taurine transporter (TAUT) in human placenta using immunocytochemistry and Western blotting, tested the hypothesis that placental protein expression of TAUT is reduced in IUGR, and investigated TAUT regulation by measuring the Na+-dependent taurine uptake in primary villous fragments after 1 h of incubation with different effectors. TAUT was primarily localized in the syncytiotrophoblast microvillous plasma membrane (MVM). TAUT was detected as a single 70-kDa band, and MVM TAUT expression was unaltered in IUGR. The PKC activator PMA and the nitric oxide (NO) donor 3-morpholinosydnonimine decreased TAUT activity (P < 0.05, n = 7–15). However, none of the tested hormones, e.g., leptin and growth hormone, altered TAUT activity significantly. PKC activity measured in MVM from control and IUGR placentas was not different. In conclusion, syncytiotrophoblast TAUT is strongly polarized to the maternal-facing plasma membrane. MVM TAUT expression is unaltered in IUGR, suggesting that the reduced MVM taurine transport in IUGR is due to changes in transporter activity. NO release downregulates placental TAUT activity, and it has previously been shown that IUGR is associated with increased fetoplacental NO levels. NO may therefore play an important role in downregulating MVM TAUT activity in IUGR.
- fetal growth
- hormones
- nitric oxide
- placental transport
- phorbol 12-myristate 13-acetate
the β-amino acid taurine (2-aminoethanesulfonic acid) is the most abundant free amino acid in a number of tissues (11) and is not incorporated into proteins. Numerous intracellular and extracellular functions have been described for taurine, including conjugation with bile acids, defense against oxygen free radicals, regulation of neuronal excitability, and involvement in cell volume regulation (11, 14, 20, 48). The main dietary sources of taurine are meat and fish, but taurine can also be synthesized de novo from cysteine in humans. However, during fetal life, taurine can be regarded as an essential amino acid, because the capacity to synthesize taurine is low or absent in the human fetus, inasmuch as it lacks the enzyme cysteine-sulfinic acid decarboxylase (11). Transplacental transfer of taurine from maternal blood to the fetal capillaries is thus the primary source of this important amino acid for the fetus. Normal fetal and neonatal development is critically dependent on the sufficient availability of taurine. It has been observed in animal experiments that taurine deficiency during pregnancy and lactation is associated with growth failure, abnormal cellular development, retinal degeneration, cardiac damage, and dysfunction of the central nervous system (19, 56, 57).
Taurine transport across the syncytiotrophoblast, the transporting epithelium of the human placenta, involves at least two fundamental steps: 1) the uptake from the maternal circulation across the microvillous plasma membrane (MVM) and 2) the transport out of the trophoblast, across the basal plasma membrane (BM) and into the umbilical circulation (3). The human taurine transporter (TAUT) has been cloned from several human tissues, including the placenta. Ramamoorthy and co-workers (50) showed that it specifically recognizes taurine and other β-amino acids and its catalytic activity is dependent on Na+ as well as Cl− with a 2:1:1 Na+-Cl−-taurine stoichiometry. TAUT has a molecular mass of ∼70 kDa, and it possesses six recognition sites for PKC-dependent phosphorylation (50). Norberg and collaborators (46) showed that the Na+-dependent uptake of taurine in BM is only 6% of that of MVM, demonstrating that the system is almost exclusively polarized to the MVM. Although not characterized in detail, Na+-independent pathways driven by the steep outwardly directed taurine gradient mediate taurine transport across BM in the fetal direction.
Intrauterine growth restriction (IUGR) can be characterized as a condition where the fetus has failed to achieve its genotypical growth potential. IUGR is associated with increased perinatal morbidity, higher incidence of neurodevelopmental impairment, and increased risk for a number of diseases in adulthood, such as cardiovascular disease and diabetes (4, 15). The altered growth pattern seen in many cases of IUGR is believed to be caused by impaired placental transport functions and blood flow (24). Norberg and colleagues (46) showed that the MVM activity of TAUT is reduced in IUGR, providing a possible mechanism for the low fetal plasma concentrations of taurine associated with IUGR (9, 12). Whether this reduction is caused by an altered number of transporters or a decrease in the affinity of the transporter for its substrate and/or a decrease in the translocation rate of the transporter-substrate complex is yet to be determined. Furthermore, it remains to be established which factors downregulate MVM taurine transport in IUGR.
Regulation of the taurine transporter has been studied primarily in cell culture models. TAUT activity has been shown to be regulated at the transcriptional or posttranscriptional level by cytokines (10, 29, 41, 53), glucose (55), nitric oxide (NO) (7, 31), p23 (18), PKC (33), and WT1 (17). PKC activation seems to be one of the major factors that downregulate TAUT activity at a posttranscriptional level (6, 16, 36, 58). Kulanthaivel and colleagues (33) demonstrated that the human placental choriocarcinoma cell line JAR expresses a taurine transporter with characteristics similar to those of the taurine transporter described in normal full-term placentas and that this transporter is inhibited by activation of PKC.
The first aim of this study was to characterize the protein expression of TAUT in the human placenta. First, we studied the cellular localization of the taurine transporter using immunocytochemistry and compared relative protein densities of TAUT in isolated MVM and BM by Western blotting. Second, we tested the hypothesis that IUGR is associated with a reduced protein expression of TAUT in MVM. Third, we investigated the regulation of the syncytiotrophoblast TAUT by measuring Na+-dependent taurine uptake in single primary villous fragments of human placenta after 1 h of incubation with different effectors. The effectors were hormones with reported alterations in fetal or maternal plasma concentrations in IUGR [growth hormone (GH), leptin, IGF-I, IGF-II, and EGF] or factors shown to regulate TAUT in other tissues [PMA, chelerythrine (CHT), TNF-α, IL-1β, IL-6, 3-morpholinosydnonimine (SIN-1), and glucose]. We also assessed the effect of the immunosuppressant rapamycin, which inhibits mammalian target of rapamycin. The mammalian target of rapamycin-signaling pathway controls protein translation in response to nutrient stimuli (22). Finally, we tested the hypothesis that PKC activity is upregulated in placentas from pregnancies complicated by IUGR.
All chemicals were purchased from Sigma-Aldrich (St. Louis, MO), except for [1,2-3H]taurine (specific activity 31–32 Ci/mmol), which was obtained from Amersham Biosciences (Buckinghamshire, UK), and TNF-α and IL-1β, which were purchased from Calbiochem (San Diego, CA).
Tissue collection.
Collection of placental tissue was approved by the Committee for Research Ethics at Göteborg University and was carried out with informed consent. Early-second-trimester placental tissue was obtained at terminations, and placentas were obtained from uncomplicated and IUGR pregnancies after vaginal or cesarean delivery at full term. The definition of IUGR was a birth weight 2 SD below the mean for gestational age with use of intrauterine growth curves based on ultrasonically estimated fetal weight (39).
Immunocytochemistry was performed as described previously (26). Briefly, placental fragments were rinsed in ice-cold physiological saline and fixed in a zinc solution. After fixation, the tissue was dehydrated through a graded series of ethanol to xylene, embedded in paraffin, cut into 4-μm sections, and mounted. Before the experiment, the slides were heated to 60°C for 20 min and allowed to cool. After removal of the paraffin in xylene and rehydration of the slides in ethanol and 0.1 M PBS, the slides were boiled in a 10 mM citrate buffer, pH 6.0, for 10 min and allowed to cool at room temperature for 30 min. Slides were washed in PBS and blocked in Blotto (normal horse serum and nonfat dry milk in PBS) for 60 or 120 min at room temperature. The slides were then incubated overnight at 4°C in a moisture chamber with a polyclonal antibody against the taurine transporter. This antibody, raised against a 14-mer synthetic peptide sequence of the taurine transporter, was diluted 1:100 or 1:200 in Blotto (16). Controls were incubated with Blotto only. Next, the slides were incubated with secondary biotinylated goat anti-rabbit IgG (Vector Laboratories, Burlingame, CA) for 30 or 45 min at room temperature. The secondary antibody was diluted 1:300 in 1.5% normal horse serum in PBS. After incubation, excess secondary antibody was washed off, and the slides were placed in 0.6% H2O2 in methanol for 10 min to block endogenous peroxidases. A Vectastain Elite ABC kit (Vector Laboratories) was used to detect the primary antibody. Slides were incubated in ABC (Vector Laboratories) for 30 or 45 min at room temperature. For visualization of the antigen, the slides were incubated with 3,5-diaminobenzidine according to the glucose oxidase method until a black reaction product appeared. The slides were then placed in PBS, dehydrated in ethanol, cleared in xylene, and mounted.
Isolation of membrane vesicles.
The microvillous and basal membranes of the syncytiotrophoblast were isolated from human placenta according to a method described previously (21, 26). Briefly, the placenta was placed on ice immediately after delivery. Vesicle preparation started within 30 min. Centrifugations were carried out at 4°C, and all other steps were conducted on ice. The placenta was dissected, and decidua, chorionic plate, and amniotic sac were removed. Approximately 100 g of villous tissue were then cut into small pieces and rinsed in ice-cold physiological saline to remove blood. Tissue was placed in buffer D [250 mM sucrose, 10 mM HEPES-Tris, pH 7.4 at 4°C, protease inhibitors (0.7 μM pepstatin A, 1.6 μM antipain, and 80 μM aprotinin), and 1 mM EDTA] and homogenized. The homogenate was centrifuged at 10,000 g for 15 min, the supernatant was collected, and the pellet was resuspended in buffer D and centrifuged again at 10,000 g for 15 min. The two resulting supernatants were combined and centrifuged at 125,000 g for 30 min. The pelleted crude membrane fraction was resuspended in buffer D, and 12 mM MgCl2 was added. The mixture was stirred slowly on ice for 20 min and then centrifuged at 2,500 g for 10 min. The supernatant, containing MVM vesicles, was centrifuged at 125,000 g for 30 min, and the pellet, containing BM, was further purified by means of a sucrose step gradient centrifugation. Finally, BM and MVM were centrifuged at 125,000 g for 30 min, and the pellets were resuspended in an appropriate volume of buffer D to give a final protein concentration of 5–10 mg/ml. Vesicle suspension was aliquoted, snap frozen in liquid nitrogen, and stored at −80°C until use.
Measurement of protein concentration.
All protein concentrations were determined according to the method of Bradford (5) using a protein assay procedure (Bio-Rad, Hercules, CA) and BSA as the standard.
Western blotting.
The proteins were separated by SDS-PAGE as described previously by Johansson et al. (26), with minor modifications. Vesicle suspension was thawed on ice and diluted in buffer D and sample buffer [8 M urea, 170 mM (5%) SDS, 0.04 U of bromphenol blue, 250 mM DTT, and 50 mM Tris·HCl, pH 6.8] to a final protein concentration of 1 mg/ml. The samples were heated to 95°C for 5 min and then cooled on ice. Fifteen micrograms of vesicle protein and appropriate prestained molecular weight markers were loaded on a 7% SDS-polyacrylamide gel. Electrophoresis was carried out at 200 V. The gels were then equilibrated in transfer buffer for 30 min under gentle agitation. Nitrocellulose transfer membranes, cut in the same sizes as the gels, were soaked in distilled water and then equilibrated in transfer buffer for 10 min. The gels were transferred overnight at 30 V. To minimize unspecific binding, the membranes were blocked in 5% Blotto buffer [0.1 M PBS, 0.1% Tween 20 (vol/vol), and 5% nonfat dry milk (wt/vol)] for 1 h. After the membranes were washed in PBS-Tween, the primary rabbit antitaurine transporter polyclonal antibody (Chemicon, Temecula, CA) diluted 1:4,000 in PBS-Tween was added, and membranes were incubated overnight at 4°C. The membranes were washed in PBS-Tween before incubation in secondary horseradish peroxidase-labeled goat anti-rabbit antibody diluted 1:1,000 (Vector Laboratories) for 1 h at room temperature. After another round of washing in PBS-Tween, enhanced chemiluminescence Western blotting detection reagents (Amersham) were used to visualize the taurine transporter bands on autoradiographic film. Relative density of the bands was evaluated by densitometry with IP Lab Gel (Signal Analytics, Vienna, VA) or Image Gauge software (version 3.45, Fuji Film). The mean density of the control bands was assigned an arbitrary value of 1, and the mean density of the IUGR group could then be compared with the control group.
Measurement of taurine transporter activity.
Taurine transporter activity was measured according to a method developed for the system A amino acid transporter (23). Tissue from newly delivered placenta was collected and dissected into small (∼5-mm3) pieces and washed in physiological saline and placed in 1:3 DMEM-Tyrode solution (135 mM NaCl, 5.0 mM KCl, 1.8 mM CaCl2, 1.0 mM MgCl2, 10 mM HEPES, and 5.6 mM d-glucose, pH 7.4) at room temperature for transport to the laboratory. Fragments were placed in fresh DMEM-Tyrode solution and further dissected into ∼1-mm3 pieces. Subsequently, the fragments were tied to one end of a silk suture, and the other end was attached to a triplicate of specially designed hooks. The villous fragments were then incubated in 1:3 DMEM-Tyrode solution (pH 7.4 at 37°C), with or without the effectors under study, for 1 h. Incubations were carried out in 4 ml of buffer at 37°C. Taurine uptake experiments were carried out in Tyrode solution only, allowing easy replacement of Na+ to obtain an Na+-free incubation buffer. In Na+-free Tyrode solution, 135 mM choline chloride was used to replace Na+. To assess the substrate specificity of the transport system mediating taurine uptake, the uptake in the presence of 1 mM β-alanine (a system β substrate) was measured. Taurine transport activity was then determined by subtracting the uptake in the control Na+-free buffer from the uptake in the medium containing β-alanine and Na+. In experiments assessing the role of Cl− for the activity of the transporter, MgSO4 replaced MgCl2 and gluconate replaced all other Cl− salts. Two sets of triplicates were studied at the same time, one incubated in Tyrode solution containing Na+ and the other in Na+-free Tyrode solution. The uptake in the Na+-free buffer represents nonspecific binding and Na+-independent uptake only. After incubation with the effectors, the fragments were washed in Tyrode solution (with or without Na+) for 2 min under constant agitation and then incubated for 20 min in Tyrode solution (with or without Na+) containing [3H]taurine (final concentration 25 nM). The fragments were then placed in ice-cold Na+-free Tyrode solution to stop the uptake. To lyse the cells, fragments were placed in distilled water overnight to release the [3H]taurine taken up by the cells into the water. On the next morning, the fragments were transferred to 0.3 M NaOH and left overnight. Scintillation fluid was added to the vials containing distilled water, mixed thoroughly, and counted in a scintillation counter. After denaturation in NaOH overnight, the protein concentration was measured. [3H]taurine uptake could be calculated as femtomoles of taurine per milligram of protein per 20 min. Taurine transporter activity was determined by subtracting the uptake in the Na+-free buffers from the uptake in the medium containing Na+.
The effect of PMA, CHT, IGF-I, leptin, GH, TNF-α, IL-1β, IL-6, EGF, rapamycin, glucose, SIN-1, and IGF-II was studied. PMA, CHT, and rapamycin were prepared in DMSO and aliquoted and stored at −20°C until use. Leptin was diluted in 15 mM HCl and 7.5 mM NaOH and stored at −20°C. GH was dissolved in 10 mM Tris buffer containing 25 mM NaCl, 0.3% BSA, and 0.015% thimerosal, pH 7.4, and stored at 4°C for ≤1 mo. IGF-I was diluted in 10 mM HCl and aliquoted and stored at −20°C. Before each experiment, PBS, pH 7.4, with 0.1% BSA was added. EGF and IGF-II were diluted in 0.2 μm-filtered 10 mM acetic acid containing 0.1% BSA. Before each experiment, sterile-filtered PBS, pH 7.4, with 0.1% BSA was added to the IGF-II stock solution. SIN-1 was diluted in water and stored in −20°C until use. All stock solutions were diluted in DMEM-Tyrode buffer to give final concentrations of 617 ng/ml PMA, 1,920 ng/ml CHT, 500 ng/ml leptin, 500 ng/ml GH, 250 ng/ml IGF-I, 600 ng/ml EGF, 20 ng/ml rapamycin, 5.4 mg/ml glucose, 620 μg/ml SIN-1, and 250 ng/ml IGF-II and then used for incubation. The final concentration of DMSO was 0.02–0.08%. Controls (i.e., no effector added) and all samples were treated with an equal amount of the solvent. IL-6, TNF-α, and IL-1β were diluted in sterile 0.1 M PBS with 0.1% BSA, aliquoted, and stored at −20°C until use. The stock solutions were then diluted in DMEM-Tyrode buffer to give the following final concentrations: 30 ng/ml IL-6, 20 ng/ml TNF-α, and 20 ng/ml IL-1β.
For most effectors, the final working concentrations were chosen on the basis of concentrations previously shown to affect taurine uptake [PMA (33), glucose (55), TNF-α (29), CHT (58), SIN-1 (7), and IL-1β (53)]; IL-6 concentration was chosen to be in the same range as IL-1β. The concentrations of the hormones were chosen to be supraphysiological compared with circulating plasma levels in humans.
PKC activity.
PKC activity was measured in MVM from normal full-term placentas and placentas from pregnancies complicated by IUGR with use of a PKC enzyme assay kit (Amersham). Vesicle suspension was thawed on ice and diluted with buffer D, pH 7.5 at 37°C, to a protein concentration of 130 μg/ml. Subsequently, 25 μl of vesicle sample were added to the assay buffer, and [32P]ATP buffer was added according to the protocol provided by the manufacturer. The samples were incubated in a 37°C water bath for 10 min; addition of a stop buffer provided in the assay system ended the reaction. The samples were counted in a scintillation counter, and the results were calculated from the specific activity of 1.2 mM Mg[32P]ATP salt.
Data presentation and statistics.
The number of experiments (n) represents the number of different placentas studied. In the uptake study, three separate villous fragments were studied for each condition, and data were averaged to obtain a value representing that placenta. Similarly, in studies of PKC activity, experiments were performed in duplicate. Values are means ± SE. Differences between groups were evaluated statistically using the t-test or repeated-measures ANOVA followed by Dunnett's test. P < 0.05 was considered significant. To determine whether the time course for taurine uptake and PKC phosphorylation was linear, a linear regression test was performed.
Clinical characteristics.
Selected clinical data for appropriate-for-gestational age (AGA) and IUGR groups used in studies of MVM TAUT protein expression and PKC activity are given in Table 1. Gestational age was not different between the groups. Fetal weight was 38% lower (P < 0.05) in the IUGR group than in controls. Similarly, placental weight was reduced by 38% in the IUGR group (P < 0.05). Furthermore, IUGR was associated with a reduced ponderal index (P < 0.05), suggesting asymmetric fetal growth.
Table 2 shows clinical data for individual IUGR pregnancies. In six of eight cases, abnormal Doppler flow patterns were registered in the umbilical artery during days or weeks before delivery. Other indications of fetal compromise in the IUGR group included oligohydramnios (1 case) and asphyxia (3 cases). One of the women had preeclampsia, which could be the cause of the fetal distress in that case. These data suggest that the IUGR group represents fetuses subjected to true growth restriction, rather than genetically or constitutionally small babies.
Zinc-fixed tissue sections of early-second-trimester (13 wk of gestation; Fig. 1A) and full-term placentas (Fig. 1B) were incubated with a polyclonal rabbit antibody directed against TAUT to determine its cellular localization in the placenta. Expression of TAUT was detected primarily in MVM of the syncytiotrophoblast in early-second-trimester and full-term placentas (Fig. 1). No significant staining could be detected in the control sections (Fig. 1, inset).
Protein expression of TAUT.
We investigated differences in protein expression between MVM and BM vesicles. Figure 2A demonstrates the presence of TAUT at ∼70 kDa in MVM. BM vesicles expressed only a faint band at 70 kDa; however, a band also appeared at ∼50 kDa (Fig. 2A). The specificity of these bands was confirmed by preincubation of the primary antibody with its blocking peptide (Chemicon), which abolished the 70- and 50-kDa bands.
We proceeded by performing Western blot analysis of syncytiotrophoblast MVM isolated from placentas of AGA and IUGR subjects (Fig. 2B). Densitometry analyses showed that the relative expression of TAUT was not significantly different between AGA (1.0 ± 0.1) and IUGR groups (1.1 ± 0.1).
TAUT activity.
Uptake of taurine (25 nM) into villous fragments in the presence of Na+ was linear up to at least 30 min (P < 0.05, n = 5, linear regression); at 20 min, 87% of total uptake was Na+ dependent (Fig. 3). These data clearly demonstrate an Na+-dependent taurine uptake in villous fragments. An incubation time of 20 min was subsequently chosen for activity measurements.
First, we determined that placental TAUT exhibits a specific requirement for Cl− and that it is substrate specific (Fig. 4A). This was done by measuring the uptake of taurine in the absence of Cl− and in the presence of 1 mM β-alanine (a system β substrate), respectively. Replacement of Cl− with gluconate and 1 mM β-alanine reduced Na+-dependent taurine uptake to very low levels (P < 0.05, n = 6, ANOVA; Fig. 4A).
Three sets of uptake experiments with different effectors were studied. In the first set of experiments, we investigated the effects of the three cytokines, IL-1β, IL-6, and TNF-α. No significant changes were observed in taurine uptake after incubation with these effectors (n = 6; Fig. 4A).
In the second set of experiments, we examined the effect of EGF (600 ng/ml), rapamycin (20 ng/ml), glucose (30 mM), SIN-1 (3 mM), and IGF-II (250 ng/ml). SIN-1 decreased TAUT activity by 35% in placental villous fragments compared with control (P < 0.05, n = 7, ANOVA; Fig. 4B).
In the third series of experiments, fragments were incubated with PMA (617 mg/ml), CHT (1,920 ng/ml), leptin (500 ng/ml), GH (500 ng/ml), and IGF-I (250 ng/ml). PMA decreased TAUT activity by 21% in placental villous fragments compared with control (P < 0.05, n = 15, ANOVA; Fig. 4C). However, neither CHT nor any of the hormones altered TAUT activity significantly (n = 15; Fig. 4C).
Incubation with the effectors did not affect the Na+-independent uptake in any of the three sets of experiments (data not shown).
PKC activity.
PKC activity in MVM isolated from normal full-term placentas was linear up to at least 10 min (P < 0.05, n = 3, linear regression; data not shown); consequently, an incubation time of 10 min was chosen. No difference was observed in PKC activity in MVM isolated from AGA and IUGR placentas: 2,311 ± 367 and 1,804 ± 312 pmol·min−1· mg protein−1, respectively (n = 8).
Taurine is present in very high concentrations in mammalian tissues, often 100-fold higher than the extracellular concentrations (49). It is not incorporated into proteins because of the β-position of the amino group, but it still has a multitude of physiological functions (11). Animal experiments have shown that an adequate supply of taurine is critical for normal growth and development of the fetus (19, 56, 57). Because of low or absent activity of cysteine-sulfinic acid decarboxylase, the rate-limiting step in the taurine synthesis pathway, taurine can be regarded as an essential amino acid during fetal life (11). Placental transport is therefore the fetus' key source of taurine.
A number of studies have suggested the presence of a single, high-affinity, Na+- and Cl−-dependent transport system selective for β-amino acids in the MVM of the polarized syncytiotrophoblasts (30, 40, 43). Norberg et al. (46) studied taurine transport in BM and showed that the Na+-dependent uptake of taurine in BM is only 6% of that of MVM, demonstrating that the system is almost exclusively polarized to MVM. The present study has, for the first time, examined the protein expression of TAUT in the placenta; the immunocytochemistry and Western blot results (the 70-kDa band) are in line with previous results showing a much higher Na+-dependent uptake in MVM than in BM.
Western blotting experiments using the anti-TAUT antibody (Chemicon) revealed two distinct bands (70 and 50 kDa) in BM. We proceeded by testing the specificity of these results by preincubating the primary antibody with its blocking peptide, which prevented detection of both bands. This indicates that the 50-kDa band is the result of specific binding of the antibody. The TAUT gene in mouse epididymis has recently been proposed to be subjected to alternative splicing (61), producing alternative transcripts, resulting in one or more proteins with shorter amino acid sequences than the 70-kDa TAUT. It is possible that the 70- and 50-kDa bands in BM are the result of alternative splicing of the TAUT gene in the syncytiotrophoblast and that the 50-kDa product is preferentially targeted to BM. When immunocytochemistry was performed, the antibody (16) showed no distinct staining in BM. This may be due to the fact that the antibody only binds to epitopes present only on the 70-kDa isoform. Because taurine is accumulated inside the syncytiotrophoblast cell by the MVM Na+-taurine cotransporter, the steep outwardly directed taurine gradient probably drives the taurine transport across BM. However, because taurine is almost completely zwitterionic at physiological pH, nonmediated transport across BM is likely to be small. The possible function of the protein represented by the 50-kDa band may be to mediate the Na+-independent taurine efflux from the syncytiotrophoblast. Interestingly, Jones and co-workers (28) demonstrated that in the polarized renal epithelial cell line LLC-PK1, TAUT in the apical membrane and in the basolateral membrane exhibits different Na+-taurine stoichiometries. Furthermore, on the basis of these studies and the detection of two potential chromosomal sites for the taurine transporter, Ramamoorthy et al. (50) proposed that the polarized placental syncytiotrophoblast expresses two different Na+-dependent taurine transporters.
Fetal plasma concentrations of taurine are known to be lower in pregnancies complicated by IUGR than in AGA pregnancies (9, 12). IUGR is associated with a distinct alteration in the MVM placental transport system for taurine. It was shown that Na+-dependent taurine uptake was reduced by 34% in MVM isolated from IUGR full-term placentas, and it was suggested that the low plasma concentrations often found in IUGR fetuses could be caused by a reduced activity of placental taurine transporters (46). Because of the critical role of taurine supply in fetal growth and development (56), information on the cellular mechanism underlying reduced placental taurine transport in IUGR is important. One possible explanation for the reduced activity in the Na+-dependent TAUT is a reduced protein expression, thus giving a reduced number of transporters. However, in this study, we have shown that the protein expression of TAUT is not altered in full-term placental MVM of IUGR babies, indicating that the reduced Na+-dependent uptake in MVM from IUGR is caused by posttranslational regulation.
The cells have two ways of controlling taurine uptake across MVM: 1) altering the number of transporter proteins in the membrane by regulating protein synthesis and degradation and 2) producing conformational changes in the protein, which alter its affinity for ligands. Molecular cloning and characterization of taurine transporters have revealed several possible phosphorylation sites (25, 35, 50, 54, 60), lending support to the possibility of transporter modulation. Han and co-workers (16) demonstrated that Ser322 on the fourth intracellular segment is the critical site for PKC phosphorylation in Madin-Darby canine kidney cell taurine transporter and that this site is 100% conserved for TAUT in all species. The reduced activity of the transporters in IUGR placentas could be caused by phosphorylation of this site, but this remains to be established.
IUGR is associated with changes in fetal and/or maternal levels or placental expression/secretion of a number of hormones and growth factors, providing the rationale for most of the effectors tested in our regulation study. Fetal levels of IGF-I (13), leptin (34), and EGF (52) are reduced in IUGR. Maternal levels of GH (13) are also reduced. IUGR placentas show higher amounts of transcripts for IGF-II than normal placentas (1). There are contradictory results concerning maternal and fetal cytokine levels in IUGR. Some studies show elevated levels of cytokines, others show reduced levels, and still others show no change in cytokine production in association with IUGR (2, 27, 51).
We proceeded by measuring the Na+-dependent taurine uptake in primary villous fragments after incubation with or without the effectors under study. The incubation time was set to 1 h, because the Western blotting data showed that MVM TAUT expression is unaltered in IUGR, suggesting that the reduction in MVM taurine transport in this pregnancy complication is not due to changes in transcription of the transporter but, rather, to changes in the activity. Our group previously validated that the fragments are structurally and functionally intact throughout the time period needed for the experiment, i.e., for up to 3 h after delivery (23). The concentrations of most effectors were chosen on the basis of concentrations previously shown to affect taurine uptake. Hormone concentrations were supraphysiological. However, for hormones produced by the syncytiotrophoblast and/or decidua, the in vivo concentrations close to the placental barrier are likely to be higher than in the general circulation.
Leptin, in the dose used in this study, has been shown to increase the activity of the system A transporter in primary villous fragments (23). However, we did not observe any effect of the hormones on taurine uptake. These findings suggest that the placental taurine transporter was not subjected to short-term (1 h) hormonal regulation.
The activity of TAUT was inhibited by the phorbol ester PMA, known to be an effective substitute for the physiologically generated diacylglycerols that activate PKC. This finding is in line with previous studies in different cell lines (6, 16, 36, 58), including the human placental JAR cell line (33). This study suggests that PKC inhibits TAUT at the posttranslational level. One hour, the time used for incubation of fragments in PMA, is likely to be too short to result in measurable effects on transporter synthesis. We speculated that an increased PKC activity in MVM in association to IUGR may contribute to the reduced taurine uptake across MVM in this pregnancy complication, but when we tested this hypothesis, no difference in PKC activity in MVM could be distinguished between AGA and IUGR groups. This indicates that PKC-mediated inhibition of TAUT may not contribute to the reduced activity of TAUT in MVM in association with IUGR.
NO is a signal molecule with diverse functions that include vasodilatation, inhibition of platelet aggregation and adhesion to the endothelium, and control of blood pressure (32). The enzymes responsible for the synthesis of NO from l-arginine are known as NO synthases (NOS) (47). Human placental syncytiotrophoblasts and villous endothelium express endothelial NOS (eNOS), and NO released from these cells is believed to be a key vasodilator within the placental vasculature (8, 44). Although there are a few reports suggesting that fetoplacental NO production is decreased in IUGR (42), most studies support an increased NOS activity in this pregnancy complication. For example, higher concentrations of plasma nitrates and nitrites (a reflection of NOS activity) have been found in fetal circulation in pregnancies complicated by IUGR (38). This may be due to the increased expression of eNOS in terminal villous vessels and stem villous vessel endothelium from IUGR placentas (45). The increased eNOS expression could be an adaptive response to the increased resistance (59) and reduced blood flow (37) in IUGR. Our results show a 35% reduction of TAUT activity after 1 h of incubation with the NO donor SIN-1. Because IUGR is associated with higher levels of NO, this might represent a mechanism for the reduced taurine uptake across MVM in IUGR. There are reports showing that long-term incubation with SIN-1 increases taurine uptake (7). However, this effect was mediated through a change in protein and mRNA expression, in contrast to IUGR, where we have shown that there is no change in protein expression of TAUT. The apparent discrepant results suggest that the mechanisms by which NO affects the taurine transporter may be different in different tissues.
Recently, Khullar and colleagues (31) showed that SIN-1 inhibits several amino acid transport systems, including taurine, in MVM vesicles. This effect could also be seen on taurine uptake when fresh villous fragments were incubated with SIN-1. Interestingly, radical scavengers were able to prevent this effect in vesicles, suggesting that the altered transport may be caused by the formation of peroxynitrite, the superoxide derivate of NO. This possibility was supported by the formation of nitrotyrosine residues in MVM, an indicator of NO production (31). It is possible that formation of nitrotyrosine residues on the taurine transporter contributes to the decreased TAUT activity after SIN-1 administration in our study.
In summary, we have demonstrated that the TAUT protein is highly polarized to the MVM of the human syncytiotrophoblast, we have shown that there is no difference in MVM protein expression of the taurine transporter between normal placentas and placentas obtained from pregnancies complicated by IUGR, and we have demonstrated that PKC and SIN-1 inhibit taurine transporter activity. Because of the high NO levels in IUGR pregnancies, NO inhibition of TAUT may decrease placental taurine transport and contribute to reduced fetal taurine levels in this pregnancy complication. We also showed by Western blot that BM contains two translation products of the taurine transporter compatible with the possibility that two isoforms of TAUT are expressed in BM. However, further studies are required to verify this hypothesis and elucidate the functional implications of this finding.
This study was supported by grants from the Wera Ekström Foundation, Swedish Research Council Grant 10838, the Swedish Diabetes Association, the Novo Nordisk Foundation, Frimurare-Barnhus-direktionen, the Åhlens Foundation, the Åke Wiberg Foundation, the Sven Jerring Foundation, and the Magnus Bergvall Foundation.
The authors thank R. W. Chesney and Dr. X. Han (University of Tennessee) for kindly donating the anti-TAUT antibody used for immunocytochemistry.
The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.
- Copyright © 2004 the American Physiological Society


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Trade Ed. Ziggurat Productions 2003
CD ISBN 1884214274 $15.00 One CDs
"I forced the last stone into its position; I plastered it up. Against the new masonry I re-erected the old rampart of bones." There's a reason Edgar Allan Poe is still widely popular after nearly 175 years. His tales are well told, timeless, and darned creepy. Jim Gallant's readings of "The Masque of the Red Death," "Silence," and "The Cask of Amontillado" do an excellent and understated job of bringing out the rhythms and meanings in Poe's richly drawn text. A subtle original musical score provides an appropriately dark background. This is a classy production and a nice introduction to some of Poe's best-known tales. B.P.
(c) AudioFile 2004, Portland, Maine [Published: FEB/ MAR 04]
Trade Ed. Ziggurat Productions 2003
CD ISBN 1884214274 $15.00 One CDs
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subset: commoncrawl


text:
GraphicRiver Grunge Christmas Texture 6366615
Photoshop \ Stock Graphic \ Backgrounds \ Miscellaneous
Christmas \ Graphic \ Designers \ Template \ Artists \ Texture \ Digital
Stock Graphic - Grunge Christmas Texture 6366615 by mKrukowski @ GraphicRiver
Photoshop \ Stock Graphic \ Backgrounds \ Miscellaneous
Grunge Christmas Texture
Great for digital artists, photographers and graphic designers
PSD Template.
- Easy to edit layered psd file
- 8 JPG files 3500×3500px each
High Resolution: Yes; Layered: Yes; Graphics Files Included: Photoshop PSD, JPG Image; Minimum Adobe CS Version: CS5; Pixel Dimensions: 3500x3500; Print Dimensions: 11x11.
Keywords: abstract, art, background, card, celebration, christmas, decoration, design, frame, graphic, greeting, holiday, illustration, invitation, landscape, merry christmas, nature, ornament, pattern, reindeer, retro, snow, snowflake, snowman, tree, vector, vintage, wallpaper, winter, xmas.


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Newsweek is Officially TheDailyBeast.com
x-posted at Zennie62.com
Just received the newest issue of Newsweek in the mail and up on top of the header for Newsweek is a little link that says THEDAILYBEAST.COM - which has never been there before. It is now official. Newsweek.com is no longer in existence and Thedailybeast.com is the new site.
The bottom of EVERY SINGLE PAGE under the page number is a link to thedailybeast.com. Every single page. And the Daily Beast Logo is everywhere.
And then there's a NewsBeast title. Pretty sure Zennie62 Media came up with that one first.
What has happened to Newsweek? The Daily Beast has taken over.
There will be a video to accompany this blog post posted as soon as it has finished loading on Youtube.


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Magic Sketchbook Trailer (2014)
01 January 2014
Fantasy Family Short Film Sara finds a sketchbook among old books in an attic room of her grandmother's home. She draws pictures and they pop up from the sketchbook.
Watch the official Magic Sketchbook 2014 trailer in HD below or find more Magic Sketchbook videos on Vidimovie.


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Happiest time of Samantha Ruth Prabhu's life - 15 photos to prove it
Submitted by NewsCrunch on Mon, 2016-12-26 21:24
Actress Samantha Ruth Prabhu is living through the happiest time of her life. If you want any proof take a peep into her Instagram page.
The actress is all set to get engaged to her longtime sweetheart Naga Chaitanya.
And it is Christmas,a time of the year she always cherishes.
Born to devout Christian parents in Chennai, she has had a religious upbringing. Samantha has always spent Christmas with her parents.
But this year, there seems to have been a change of plans. Going by her Instagram photos, she seems to have celebrated Christmas a week early before dashing off for quick beach-side vacation with Naga Chaitanya.
Samantha posted a photo in bikini and that has gone viral.


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Discussion in 'Punto 1.3 MJD' started by RajiniHasan, Dec 4, 2012.
Welcome to the family buddy and congrats for your new ride.
Firstly welcome to the FIAT/Punto family.
Its a great decisson that you followed your heart, be assured that this will keep your mind happy too
All the best for the PDI and delivery of your ride.
Welcome to TFI enjoy your ride am sure u Will never regarde your decision.
Congrats on your decision!
I was in a similar fix exactly a month back and it was the Italian babe that won me over!
The Punto pips the Polo in almost every way. Even if its a decision from the heart, it would still keep your mind pleased
Waiting for your delivery experience and review.
And its great news that DSC has taken up the FIAT dealership at Chennai!
Congratulations! exactly the same reasons I went for punto !
Hi Shanmugam, its from VST
Congratulations, RajiniHasan.
Had did my PDI today.. It was Oct 2012 Manufacturing. Though my car was placed in open place mixed of weather these days still looks good. I was not able to check on Odo meter. Sales person said battery will be unfitted for safety purposes. Hoping everything is good. Have taken few snaps..
Also cheers to Chennai Fiatians... DSC has finally opening up its exclusive showroom "RDC FIAT" in CIT Nagar, next to its Hyundai showroom on 9th Dec... wow.. 2 days to go..
your beauty looks so cool and great !! indeed it is a very good news that fiat showroom going to opening up in another 2 days....
Congrats and welcome to the family . Please do check the ODO for the tell tale 'H'.
Though you have posted stating that the sales person said that the battery will be unfitted (guess you meant disconnected), your picture of the engine bay tells a different tale
Separate names with a comma.


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Have you been suffering from sinusitis that results in headaches and blocked nose, for a while now? If yes, then here is a powerful home remedy for sinusitis that has shocked doctors!
If you are someone who often keeps getting infected by sinus infections, then you will definitely know how painful, uncomfortable and frustrating it can be, right?
Blocked nose, headaches, heaviness in the head - these symptoms associated with sinus infections are bound to hamper a person's daily life to a considerable extent.
When a person has sinusitis, he/she could find it very difficult to spend time in the AC, consume cold foods and even a little dust can irritate their nasal tract!
Sinusitis can be described as a condition in which the cavities around the nasal passages become inflamed and congested with mucous, produced by infection.
Some of the main symptoms of sinusitis are pain in the forehead, cheeks, nose, etc., headache, coughing, nasal congestion, difficulty in breathing, loss of smell, fatigue, fever, etc.
So, learn how to prepare and use this remedy to treat sinusitis, here!
Recipe To Prepare The Remedy
Ingredients Required :
- Oregano Oil - a few drops
- Turmeric - 2 teaspoons
- Hot Water - 1 cup
This natural remedy to treat sinusitis can work wonders, specially if you avoid consuming oily and cold foods.
You must also avoid spending a lot of time in the AC and avoid being exposed to pollution, for this remedy to work effectively.
The mixture of oregano oil, turmeric and hot water works to reduce the inflammation of your sinus duct and also dissolve the mucus accumulated in the sinus duct, thus reducing the pain and congestion.
Method To Prepare And Use The Remedy :
- Add the suggested amounts of oregano oil and turmeric into a cup of steaming hot water.
- Now, stir the ingredients well to form a mixture.
- Your remedy is now ready for consumption.
- Consume this remedy every night, after dinner, for at least a month.
Try this natural remedy at home and give us a shout out if you liked it!


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Skip to comments.In order to exonerate the W.H. NSA should release all meta
Posted on 05/11/2014 6:52:15 PM PDT by big bad easter bunny
Time to exonerate the White House, lets have the NSA release the meta data on the IRS, Bengazi and Fast and Furious, this with hunt must end, there is not a smidgen of corruption, the most transparent administration can easily prove it!
I intended on witch not with.
I don't think they can provide the answer to, "where was Barry and what was he doing during the attack," and "who gave the stand-down order"?
meta data?
Sure they will. After its edited.../s
Honestly, does anyone think have NSA or CIA or SS speaking on one's behalf could improve public perception?
Disclaimer: Opinions posted on Free Republic are those of the individual posters and do not necessarily represent the opinion of Free Republic or its management. All materials posted herein are protected by copyright law and the exemption for fair use of copyrighted works.


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Velvet Film is a production company created by Raoul Peck in 1986 in Germany. It is currently based in France, Haiti and in the United States of America.
Company presentation
Filmography (5)
The Young Karl Marx
I Am Not Your Negro
Fatal Assistance
(2013 - Post Production)
Murder in Pacot
(2013 - Upcoming release)
Moloch Tropical
(2009 - Upcoming release)
Sorry, your search returned no results.
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Staff (2)
To access public contact details in the UniFrance directory, please log in (or register on our website).


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Cadillac Of South Charlotte Body Shop
Featured Business
ABRA Glass Charlotte10421 S Tryon St.
Charlotte, NC 28273
4.2 miles away
"John said they were excellent."
Review for Cadillac Of South Charlotte Body Shop
Gregory B. on 05/16/2011
Gregory said he had a horrible experience because when he picked up the vehicle, it was closing time and the person at the counter was hurrying him up because she had to be some place. He paid $8,000 and customer should not be told this especially when paying this much. The vehicle was very dirty and it should be detailed, there was paint on the front leather seat, the floor mat had rust and grease. He said there were greasy fingerprints on the headliner, disposable gloves on the back floor, body trim, tools and bolts in the back as well. There was no promised time. He had to speak to the business manager to get this fixed and he feels he needs to be compensated for gas, time and time off from work because of this horrible experience and would not recommend them to anyone. He also said the rear view mirror fell off.
Category: Collision
Service Date: 05/06/2011
Review Created: 05/16/2011
This review was collected via phone interview by Customer Research Inc.


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From today's Daily News:
MAYOR NUTTER and City Council President Darrell Clarke have reached a compromise on the renovation of JFK Plaza (a/k/a LOVE Park) after weeks of sparring over the issue.
Clarke agreed to allow plans for the sale of the garage beneath the plaza to advance in City Council, a transaction Nutter's been pushing for since the city garnered a more generous offer for it than was expected.
In exchange, the mayor agreed to "dedicated space for [food] concessions" at the site, while seeking participation from the public and the private sector in studying ways to maximize revenue.
"After a number of conversations between Council President Clarke and myself, the opportunity to lower the ultimate taxpayer cost, supplanted by corporate philanthropic or some amount of revenue-generation at the site - that combination is just a good idea," Nutter said.
The city originally asked for $20 million for the 810-space garage, but the highest bidder - Chicago-based InterPark LLC - eclipsed that figure, offering $29.6 million.
Clarke and Nutter looked triumphant announcing their agreement during a news conference in front of Robert Indiana's iconic "LOVE" sculpture yesterday afternoon.
"The refurbishment of this park is long overdue," Clarke said.
"This was not an adversarial relationship, although it may have appeared to be that way in some of the press accounts, but the reality is that we have been working together over the last several months. I'm excited about the prospects of having a well-balanced approach to redoing this park."
Last month, Clarke entertained a proposal for seven restaurants on the park apron - the revenues of which he envisioned would pay for the renovations. But backlash from community groups may have struck a chord.
"This is good news, but it remains to be seen what the details are," said Lauren Bornfriend, executive director of the Philadelphia Parks Alliance, who represents one of the more vocal objectors to the restaurants proposal.
"Our assumption is that, with the public process, the plan they arrive at will look very different. We'll continue to be vigilant to ensure city and state laws that protect parkland are adhered to."
Nutter said he wants to see food or restaurant concessions balanced with enhanced green space for the park. He said the city would apply for $3 million from the commonwealth's Redevelopment Assistance Capital Program.
"Remaining costs would be funded either through proceeds from the sale of the garage beneath the plaza, through the city's capital investment program or both," said Nutter, who invited private corporations and foundations to share input.
"We welcome institutions to step forward immediately if they are interested in this project," he said.


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All Camps
filters camps by age:
Medieval Week
Learn what it takes to become a Knight of Kern by training in the arts of swordsmanship, archery, and much more! At the end of the week participate in a skills competition against your fellow knights and see if you have what it takes to earn your seat as a Knight of the Round Table!
Ranger's Apprentice
Immerse yourself in the world of Ranger's Apprentice! This week is fully-themed, and you will be instructed on horseback riding, trail riding, archery, tracking, survival skills, stealth and Ranger missions. If you love Ranger's Apprentice, this officially-sanctioned camp will test your strength, loyalty and commitment. Defend the kingdom from Morgarath's Wargals, Skandian Raiders, and the fearsome Temujai warriors. Come as an Apprentice. Earn your Silver Oak Leaf. Become a Ranger. ...
Harry Potter Camp
For many years, Camp Kern has played Harry Potter as a Big Game. Multiple Wednesday nights each summer are spent helping Harry and his friends save Hogwarts from Voldemort. These years have been very informative and fun, but for some, its not enough! Now, live the magical world of Harry Potter for an ENTIRE WEEK at Camp Kern! From the moment you arrive, until you check out on Saturday, you will cease to be a Muggle, and will enter the Hogwarts School of Witchcraft and Wizardry as a studen...
Percy Jackson Week
You've always felt that you didn't quite belong. Things never seemed to look the same to you as they did to other kids. One of your parents seems suspiciously "god-like". You can read Latin. You are a great candidate for Camp Jupiter! Come spend a week at camp, training your body and mind into the hero's physique you were build for. Daily quests, monsters to vanquish, and a god-written destiny await all who answer the call of heroes! &nbs...
Star Wars Week
Are you prepared to embark on the greatest adventure ever seen in a galaxy long ago and far away? There is war brewing between the Republic, Clones, Trade Federation, Outer Rim Planets…and there have been reports that the Sith are reemerging. Join forces with the greatest heroes of the universe and take down the darkest evil ever faced. Your week will train you in the ways and honor of the Jedi, while challenging you with the nightly missions. Gather your courage and...
Advanced Ranger's Apprentice
For those who love Ranger's Apprentice camp and are looking for more, Advanced Ranger's Apprentice is for you. Build on your Ranger skills in horseback riding, archery, and stealthing while taking on even more challenging missions. This week will test Ranger's Apprentices to see if they have what it takes to become a Ranger. Previous participation in Ranger's Apprentice Camp is recommended for Advanced Ranger's Apprentice. Advanced Ranger's Apprentice can be combined with Rang...
Pokemon Week
Give your camper the chance to train under the best Pokémon Masters the world has ever seen! Learn the skills necessary to become a Pokémon Trainer, catch and battle with Pokémon, and even learn what it takes to beat the Elite 4! Don't miss out on this opportunity of a lifetime!


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Know more about this business than we do? Cool! Please submit any corrections or missing details you may have.Help us make it right
Be the first AL.com user to add photos or videos of Boulder Construction Co I
Boulder Construction Company Inc is located at 6587 Dauphin Island Pkwy, Mobile, AL. This location is in the Alligator Bayou neighborhood. This business specializes in Home Repair & Improvement.
Boulder Construction Company Inc can be found at Dauphin Island Pky 6587. The following is offered: B2B Contractors. The entry is present with us since Sep 8, 2010 and was last updated on Nov 12, 2013. In Mobile there are 208 other B2B Contractors. An overview can be found here.


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Lenox Poppies on Blue Kitchen Clock
Sold for:
Start Free Trial
or Sign In to see what it's worth.
- Sold Date:
- Source: eBay
This is the Poppies on Blue Clock by Lenox. It runs on one AA battery. Works perfectly. It is completely free from cracks or chips, but is not in the original box. From a smoke-free, pet-free home. If you like Poppies on Blue, check out my Creamer and Sugar Bowl! Always happy to combine shipping!Due to the very breakable nature, I'm requiring insurance on this one. Even though I bubble-wrap them to death, I don't want anyone to lose money! Thanks for looking!


text:
Review: Ignition Switch Cover ring
Ignition Switch Cover Ring White by Indesit
£11.48 Only 1 Left in Stock
The part fits well but is very expensive for what it is hence 2 stars. I also ordered 'Clear Cover - Ignition Switch part C00039633' which came with a black cover ring.
Very easy to fit
Fitted to cooker hob Ariston System 2000 - A2030
24 Oct 2011 by Anonymous
View all Reviews


text:
You can leave your hat on... Actor Neil Patrick Harris strips for latest cover of Rolling Stone
Neil Patrick Harris is going by the 'Full Monty' mantra 'You can leave your hat on...' in his latest cover shoot.
The actor appears on the cover of 'Rolling Stone' magazine with just a dickie bow on and a strategically-placed top hat.
The 'How I Met Your Mother' actor spoke about the impact coming out as gay has had on his career and said he got 'more opportunities' than ever.
"Once all the cards were on the table, I got more opportunities than ever," he told the magazine.
"Some actors don't get hired because you can't look into their soul and see what they're like, because they're kept guarded."
He also said that revealing his homosexuality to his family was more difficult than coming out to the public and his fans.
Harris's brother Brian said that Neil came out to the family 'over three conversations;
"It was three conversations," he said.
"First he goes, 'I just don't think I'm going to date, really, for a while.' And he had some bad luck with the girls he dated early on. Probably because he was gay! And then we had a different conversation about how he likes girls, but he kind of likes guys too. And we had another conversation after that: He likes guys."
Harris is currently on stage at Broadway, starring in the title role of Hedwig and the Angry Inch.
And he said it took him a while to adjust to playing the role of the effeminate Hedwig."I didn't think he could pull off the femininity of [the role]," his fiance David Burtka said.
"This is such a stretch for him. In his day-to-day, he's not a very feminine guy."
Online Editors


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Rise in Brits heading to 'safe' Spain DESPITE jihadis' 'wanting to conquer country'
BRITS booking up holidays to Spain are up 30% – but sun seekers are flying their families straight into the firing line, it has been warned.
Travel firms are rubbing their hands with a recent surge in bookings to the "safe" havens as fearful Brits shun known ISIS targets such as Turkey and Tunisia.
But terror experts have warned that the threat to Spain still remains high despite a flurry of bookings to the popular Costa resorts.
Tom Wilson, a Middle East specialist at the Henry Jackson Society said many ISIS militants are still looking to make there way to Spain for unfinished business.
Speaking to the Daily Star Online, he said: "For one thing the country's proximity to North Africa might make it easier to reach for Jihadists coming across from particularly troubled places such as Tunisia and Libya.
"Furthermore, it is also true that Islamist militants such as Islamic State do speak of wanting to reconquer Spain on the grounds that this territory had been Islamic during the Middle Ages.
"Nevertheless, there is quite a gap between what Islamic State say they wish to do, and what they are actually able to do in practice."
He added though: "Generally speaking, Spain should not be considered any more dangerous than other parts of Western Europe."
Despite his reassurances, many neighbouring countries such as France and Belgium remain on their highest terror threat levels in recent years following a spate of deadly attacks by ISIS terror nuts.
The UK currently has a severe terror threat level - the fourth of five tiers.
And fears from other factions remain rife with MI5 raising the threat level from Northern Ireland-related terrorism just this month.
The worries at home have opened the doors for Brits to getaway to enjoy some sun abroad - with Spain and Portugal being two of the most popular destinations for bookings.
The Association of British Travel Agents (ABTA) has encouraged sun seekers to consider the two countries on the Iberian peninsula despite the Foreign Office's red alert.
It claimed bookings to Spain have since risen by 26% – and Portugal 29% – warning that holidays are on the verge of running out.
Travel analysts have warned that Brits may face paying a premium as the booking boost pushes up hotel and flight prices.
Over 12 million Brits travel to Spain every year and many own holiday homes in the likes of Benidorm, Marbella and Fuengirola.
Nonetheless, Spain has been a hotspot for extremism for centuries – and Islamic territories Ibiza and Magaluf are prime targets.
Daily Star Online revealed last year how summer could be the worst time to travel to the country.
"The country's proximity to North Africa might make it easier to reach for Jihadists coming across"Tom Wilson, Middle East specialist
Top security specialist Will Geddes warned that Brits flocking to the Costa were picking a destination that was "not the safest" and a "difficult call" for those with families.
This is because the spontaneous nature of lone wolf attacks makes it particularly difficult for countries to defend themselves.
He said: "The hardest problem is that the framework of IS is to incentivise the lone actor".
Earlier this week, it emerged how Spanish authorities were ramping up further security at congested beach resorts amid fears of an imminent attack.
Brits abroad will now be forced to hand over their passport to prove their identity if quizzed by a Spanish officer in a stop and search initiative.
Refusal to hand over ID is now a criminal offence, the Foreign Office warned on its website.
On its website, the Foreign Office's Spanish travel advice states: "There is a high threat from terrorism.
"Attacks could be indiscriminate, including in places visited by foreigners.
"The Spanish authorities take measures to protect visitors, but you should be vigilant and follow the instructions of the local authorities."
"There is considered to be a heightened threat of terrorist attack globally against UK interests and British nationals, from groups or individuals motivated by the conflict in Iraq and Syria. You should be vigilant at this time."
Those planning on making the plunge should read these seven tips to stay safe while abroad.


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Thanks to everyone who came to visit us at the Cincinnati Boat Show! The CortiBalm Challenge was a huge success, with over 100 tubes swapped. Interested in taking the CortiBalm Challenge yourself? Visit our site to find out how YOU can get a free sample today.
Monthly Archives: January 2015
LIVE from Cincinnati: Take the CortiBalm Challenge
Ready to take the CortiBalm Challenge? We're live at the Cincinnati Boat Show, Booth 1310, giving a FREE tube of Dr. Dan's CortiBalm to anyone willing to trade in their existing lip balm.


text:
Digitally printed in the UK
100% Linen
Printed width 137cm,
Square repeat, 37cm H x 45.7cm W
Cigarette tested and rub test 16,500 martindale suitable for curtaining and light upholstery.
Greenfinch Fabric
Lorna's love of British birds continues with the gorgeous Greenfinch design. A visitor to gardens in both city and country, the vivid green bird is seen here in flight against a springtime backdrop of apple blossom.
Part of Lorna's Bloom collection which celebrates the array of small wildlife Lorna has photographed whilst admiring the flowering plants and shrubs in British parks and gardens.
See this print also as a lampshade, wallpaper, cushions, plant pots and make up bags
All items are tissue wrapped before postage, making them a perfect gift for you, or someone else. If this is a present, we can provide and write a card. Please purchase the card via the website at the same time and leave us a note on the checkout form.
Price per linear meter. If more than one meter is ordered, fabric will be supplied in one continuous length.
Postage £4.95 or Free postage on orders over £75.
International and next day delivery available. See more on delivery and returns or please use the delivery calculator at check out.


text:
New upward movement is possible.
On the 4-hour chart the pair XAU/USD tested the correction level and 23.6% arc for a short-term trend several times. Currently, the price is heading towards a strong support level 1164.10. Should the level be broken through, the price may continue going down to 1148.90 (0.0% correction level for a medium-term trend). If the price bounces off this level and manages to overcome 23.6% arc, the pair is likely to grow towards 1173.40 (23.6% correction level, crossover with 38.2% arc), 1179.00 (38.2% correction level, crossover with 50.0% arc), 1183.75 (50.0% correction level, crossover with 61.8% arc). Given this, 1183.00-1185.00 is a strong resistance where 50.0% and 23.6% corrections for short and medium-term trends match.
Trading tips
If the price breaks through 23.6% arc and consolidates above 1167.20, open long positions with the targets at 1173.40, 1179.00, 1183.75 and stop-loss at 1164.50.
Alternative Scenario
The breakdown of the level 1164.10 would allow the price to fall towards 1148.90. Stop-loss should be placed at 1167.00.


text:
October 6, 2010 by staff
Dresser, Dresser Wayne, a global innovator of fuel dispensers and technologies, announced today that its iX Pay secure payment solution is the worlds first solution to receive the Payment Card Industry PIN Security transactions (PCI SCP) certification for version 3.0 encrypt a PIN (CPE) devices keyboard. While many competitive payment solutions for retail fuel are still approved only to 2004 standards, which were based on the payment practices of security attack identified more than six years, Dresser Wayne important victory that allows fuel retailers with the bond payment and advanced the most common. PTS PCI 3.0 compatible iX Pay secure payment solution helps protect consumers and retailers increasingly sophisticated methods of fraud data. It also provides industry-leading equipment investment protection through compliance with 2020, and even with upgrades.
This historic announcement further demonstrates Dresser Wayne's commitment to technological leadership in secure payment. Dresser Wayne was the first to propose a solution PCI 2.0-compliant EPP on the world market in 2007, and continues to lead the industry in technological advances with other similar developments such as the recently announced its mode Limited functionality (AMT) included in the iX Pay secure payment solution. Capacity first of its kind, AMT maintains functionality for secure processing of payment transactions non-PIN, even if the EPP is disabled as a result of alteration.
Threats to security of payment are increasingly at an alarming rate. As an industry, we must go beyond meeting the minimum criteria for compliance. We need to stay ahead of the curve in protecting consumers and businesses against identity theft and financial crimes, said Tim Weston, Dresser Wayne, product manager of payment technologies. Unlike its competitors struggling to put version 2.0 of the solutions on the market before the PCI specification retires, weve designed our security solutions integrate the latest technologies in data protection payment, and to anticipate regulatory changes rather than react.
In addition, Dresser Wayne has received the first world championship without PCI Payment Terminal (UPT) version 3.0 certification for the Ovation, Tokheim Vista and fuel prime. Consequently, fuel retailers can benefit from enhanced security for transactions by credit card and debit cards using a PIN. Configurations PCI UPT-TSP approved based Dresser Wayne's newly released Secure reader magnetic stripe card that allows you to encrypt payment data from the time a customer slides his card. As the companys current security drive hybrid EMV smart card, magnetic stripe reader transmits data securely between the components of the terminal, and is capable of supporting financial networks from end to end data encryption.
Financial data theft is a serious problem for all retailers, but for C-store retailers with fuel dispensers on the outside available to the public at all times, the problem may be compounded by a series of factors. It must be approached from many different angles, Weston said.
The solution iX Pay secure payment incorporates a multi-level approach to data protection, including the algorithms of Triple-DES encryption; built-in keyboard protects privacy, whistleblowing mechanisms on reactive and unauthorized protective device abduction, and many other features innovative safety. Its keys travel has the added benefit of preventing the use of skimmers data from the keyboard.
In addition, it is built on the platform technology upgrade iX, a single global foundation that supports various configurations to meet regional needs as well as additional security enhancements, which can minimize the costs of compliance continues with ever changing standards of security of payment. Characteristics iX Pay secure payment solutions are not robust security PTS only PCI 3.0-compliant, but they have also achieved certification to other major global standards for security of payments, such as 4.2 for EMV payment cards smart Canadian Interac 2.0 and pan-Nordic Card Association (PNC) regulations UPT.
To learn more about the solution iX Pay secure payment, visit the Dresser Wayne NACS Show 2010 in Atlanta, GA, USA, October 58, booth 537.
About Dresser, Inc.
Dresser, Inc. is an industry leader worldwide in providing highly engineered infrastructure products for the global energy industry. Leading brand names within the Dresser portfolio include Dresser Wayne retail fueling systems, Waukesha engines, natural gas, Masoneilan control valves, Consolidated pressure relief valves, and Roots blowers and rotary gas meters. The company has manufacturing facilities and customer service located strategically worldwide and a sales presence in over 150 countries.
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Home > Tags: matlab (Total 232 Records)
'Applications from Engineering with MATLAB Concepts' ed. by Jan Valdman
Computer Science By:Jan Valdman 2013-00-00 00:00:00
The book presents a collection of MATLAB-based chapters of various engineering background. Instead of giving exhausting amount of technical details, authors were rather advised to explain relations of their problems to actual MATLAB concepts. Whenever possible, download links to functioning MATLAB c matlab concepts, actual matlab concepts, matlab codes, various engineering background, lc-ms/ms fragmentation data
Introduction to Nonlinear Optimization: Theory, Algorithms, and Applications with MATLAB
Algorithms By:Amir Beck 2014-10-27 00:00:00
This book provides the foundations of the theory of nonlinear optimization as well as some related algorithms and presents a variety of applications from diverse areas of applied sciences. The author combines three pillars of optimization-theoretical and algorithmic foundation, familiarity with vari nonlinear optimization, convex optimization chapter, convex sets chapter, convex functions chapter, convex set chapter
Tags: optimization application theory algorithm nonlinear matlab
Primary MATLAB for life sciences : guide for beginners
Computer Science By:Burstein, Leonid 2013-00-00 00:00:00
This e-book provides readers a short introductory MATLAB course oriented towards various collaborative areas of biotechnology and bioscience. The text concentrates on MATLAB fundamentals and gives examples of its application for various problems in computational biology, molecular biology, biokineti short introductory matlab, life sciences, primary matlab, matlab fundamentals, various collaborative areas
Digital Signal Processing for Wireless Communication using Matlab
Digital Signal Processing By:E.S. Gopi 2015-08-10 00:00:00
This book examines signal processing techniques used in wireless communication illustrated by using the Matlab program. The author discusses these techniques as they relate to Doppler spread, Delay spread, Rayleigh and Rician channel modeling, rake receiver, diversity techniques, MIMO and OFDM based signal processing, division multiple access, signal processing techniques, binary phase shift, quadrature phase shift
Tags: signal communication digital processing wireless matlab
Variational Regularization of 3D Data: Experiments with MATLAB
Programming By:Hebert Montegranario, Jairo Espinosa 2014-00-00 00:00:00
Variational Regularization of 3D Data provides an introduction to variational methods for data modelling and its application in computer vision. In this book, the authors identify interpolation as an inverse problem that can be solved by Tikhonov regularization. The proposed solutions are generaliza variational regularization, 3d data, regularization theory, variational methods, necessary theoretical background
'Image Processing in Optical Coherence Tomography using Matlab' by Robert Koprowski, Zygmunt Wrobel
Digital Signal Processing By:Robert Koprowski, Zygmunt Wrobel 2013-00-00 00:00:00
The monograph comprises proposals of new and also of known algorithms, modified by authors, for image analysis and processing, presented on the basis of example of Matlab environment with Image Processing tools. The results are not only obtained fully automatically, but also repeatable, providing do tomographic eye image, anterior eye segment, optical coherence tomography, eye segment analysis, random contour analysis
Tags: image optical processing matlab tomography coherence koprowski zygmunt wrobel
Introduccion Rapida a Matlab Y Simulink Para Ciencia E Ingenieria / Quick Introduction to Matlab and Simulink for Science and Engineering (Spanish Edition) [Spanish]
Digital Signal Processing By:Gil Rodriguez 2003-11-19 00:00:00
MatLab (MATrix LABoratory) comenz siendo un programa interactivo de anlisis integrado, especializado en clculos matriciales. En muy poco tiempo tuvo una gran difusin, facilitado por su potencia de clculo y facilidad de uso. La primera versin de MatLab, de xito generalizado, se lanz el 3 de febrero spanish edition, matlab dispona, matrix laboratory, signal processing, introduccion rapida
Tags: engineering science matlab spanish simulink para introduccion rapida ciencia ingenieria
Accelerating MATLAB with GPU Computing: A Primer with Examples
Computer Science By:Jung W. Suh 2013-12-16 00:00:00
Many developers want to increase the speed of their MATLAB prototyping and algorithm development by leveraging the distributed parallelism of graphics processing units (GPUs), but lack the necessary experience in C coding or computer architecture. Accelerating MATLAB with GPUs offers a primer on usi matlab, real-world projectsaccelerating matlab, matlab codes, matlab prototyping, gpu computing
Real-Time Digital Signal Processing from MATLAB to C with the TMS320C6x DSPs, Second Edition
Digital Signal Processing By:Thaddeus Baynard Welch III 2011-12-22 00:00:00
Mastering practical application of real-time digital signal processing (DSP) remains one of the most challenging and time-consuming pursuits in the field. It is even more difficult without a resource to bridge the gap between theory and practice. Filling that void, Real-Time Digital Signal Processi digital signal processing, real-time digital signal, real-time dsp, real-time dsp applications, real-time dsp projects
Computer Programming with Matlab
Programming By:J . Michael Fitzpatrick, Akos Ledeczi 2013-00-00 00:00:00
The primary purpose of this book is to teach computer programming to those with little to no previous experience. It uses the programming system and language called MATLAB to do so because MATLAB is easy to learn and, at the same time, is an extremely versatile and useful programming language and pr matlab, matlab programs, matlab tutorial, useful programming language, general-purpose languages.computer programming
Tags: computer programming matlab


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Twelve and Holding Trailer (2005)
"Do You Know Who Your Kids Are?" 11 September 2005 Drama 90 mins
Three pre-adolescents with virginal and eager-of-signals naivety, hit headlong against monstrous and inhospitable loneliness, in the incandescent fire among the ashes of their own inocence.
Watch the official Twelve and Holding 2005 trailer in HD below or find more Twelve and Holding videos on Vidimovie.


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The End of a Mystery Trailer (2003)
Joaquin comes back to Granada in the eighties trying to find out about something happened when he was a child and the Spanish Civil War was going on. He helped an unknown man who survived after being executed. He finds the man, Galapago, who is now quite old, poor and with almost no memory. Joaquin takes care of him and finds hints that point to Galapago as Federico García Lorca.
Watch the official The End of a Mystery 2003 trailer in HD below or find more The End of a Mystery videos on Vidimovie.


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Right now, we are at the start of a massive migration that will utterly rewrite the relationship of an enterprise to its data but the worrisome bit is that even Cloud insiders admit there is still an awful lot that needs to be known about clouds. This extends right down to dollars and cents. Pricing is a moving target, admits Susie Adams, CTO of Microsoft Public Sector, which is involved in helping roll out Microsofts Cloud offerings.
The fact that Cloud pricing is vague probably doesnt surprise you, but the fact that Microsoft is a Cloud player just might. Accept this: around about now, almost every major IT provider claims to have some kind of "Cloud" offering because they all see what the next big thing is going to be. So, whos on first? What are they doing? For how much? Read on for the a list of whats happening in the Cloud right now.
From Hotmail to X-box Live, Microsoft has staked out its place in the clouds, but Adams wants to point a finger at the companys industrial-grade offerings, too. We havent talked much about it but we are involved, said Adams, who adds: Weve been in this space for many years. We offer a lot of enterprise-level services in the Cloud.
Live Meeting and Azure Services, a developers tool that goes head-to-head with Google App Engine, lead the list of public clouds from Microsoft, but the company also is busy creating custom clouds for government agencies and for the private sector. Microsofts big bet is that the first Cloud offering to take off will be email (Its become a pain point for many customers, said Adams) and Microsoft believes they will gladly turn over the increasingly costly aggravation that email has become. From there, Microsoft hopes to coax customers to buy more services and there is plenty of confidence this strategy will work.
By any measure, IBM is a 900-pound gorilla in the Cloud space and it has every intention of staying a leader, said Dennis Quan, director of Development, Autonomic Computing, IBM Software Group. IBM already operates 13 Cloud computer competency centers around the world (from India to Brazil to South Africa and Silicon Valley). But, insists Quan, Cloud is not one size fits all and at IBM the strategy going forward is to work with potential customers to identify what they want to move to the Cloud, what they want to hold in-house, and at what price. We see a hybrid model for Cloud services predominating, said Quan.
Perhaps even more exciting, IBM sees a growing market for in-house, private clouds. The idea may seem contradictory on its face, but not to Quan, who elaborates: The customer will be able to establish a Cloud inside its own four walls and, he suggests, this customer also will benefit from access to more robust computing solutions at what will amount to discount pricing. That just may be a winning formula in the present economic environment, according to IBM.
Online retailer Amazon, from the early days of Cloud computing, has been aggressively hunting customers and, to Amazons credit, it offers some of the clearest pricing in the industry. Sign up for Amazon Elastic Compute Cloud (EC2) and you can access, on demand, a high capacity CPU running Unix or Linux for $.80/hour. A similar computer running Windows is $1.20/hour. Amazon also has crafted a deal where it is running IBM apps inside its Cloud. And it offers vast data storage in the Cloud, via Amazon Jungle Disk; at nominal charges. Amazon increasingly figures into all conversations about adopting Cloud strategies, particularly inside SMBs. Of course, in addition to ad hoc, a la carte pricing, Amazon offers a range of subscription deals for customers with continuing needs.


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MOVIE SYNOPSIS [ General Audiences ] With direction of Michael Staal Dinesen, this film in English (original language) was presented in 2015. This film, a creation of Michael Staal Dinesen, was produced in Denmark. You can watch Svindlerne online on TV cable or theaters with original audio in Danish at different dates. This film premiered in theaters in 2015.
Rating: 5 by 10 users./
Runtime: 58 minutes
Kristian Mogensen and Jakob Bilberg Nasser.
58-minute film (full movie) directed by Michael Staal Dinesen.
The original audio of Svindlerne has dialogues in or Danish, it was produced in Denmark.
It premiered in cinemas or television in 2015.
Screenplay written by Michael Staal Dinesen.
Michael Staal Dinesen.
Ellen Riis.
This film was produced and / or financed by Basmati Film and Staalfilm.
Composed by Thomas Richard Christensen.
Staalfilm and Basmati Film.
Without commercial breaks, the full movie Svindlerne has a duration of 58 minutes; the official trailer can be watched in streaming on the Internet. You can watch this full movie free in English or Danish in movie TV channels, renting the DVD or with VoD services (Video On Demand player, Hulu) and PPV (Pay Per View, Netflix).
See the page of this full movie, Svindlerne in FULLTV Movies.


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Flexsys fined for 'reckless' storage which led to major effluent spill
Chemical manufacturer Flexsys Rubber Chemicals has been fined £10,000 for a spill of more than 200 tonnes of acidic effluent from its Ruabon works into the Dee, a major water supply river. The court attributed the incident to the company's "reckless" storage of corrosive effluent in tanks meant only for water.
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2601 N. Howard Street
Baltimore, MD 21218
Free Wifi ALL DAY!
Mon-Fri: 6am–7pm
Sat: 8am–7pm
Sun: 8am–5pm
Spicy Golden Milk has arrived at Charmington's!! Our syrup is made in-house, from scratch, and includes coconut oil, turmeric, and other spices. It's made with almond milk, but can be prepared with any milk you'd like!
Holiday hours
December 24th 8:00 AM-2:00 PM
December 25th – January 1st CLOSED
January 2nd resume normal hours
But why?
Make your holidays easier with our delicious custom baked cookie tins!
Located between Charles Village and Remington (more specifically on the corner of Howard and 26th Streets) in Baltimore, Charmington's is a coffeehouse and café serving up awesome food and beverages to its customers.


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How to describe this epic masterpiece? I've spent the last month compiling a hillarious collage of President Obama and Admiral Adama comparisons. Over eighteen seperate mini-joke/comic/things all rolled into one sprawling bizarre competition that ends with the two degenerating into insults. And it'll take you less than a minute to read. *worth it*
Saucy dangerous unpredictable Ellen Tigh has returned to the fleet only to discover - dum dum dum! - Tigh has knocked up a Six and Ellen's none to pleased. Baltar struggles for control of his culthood and the Allspark is causually mentioned.
Anders' expostion button goes off and the entire history of the humans and cylons is downloaded to us viewers in an amazing tour de force. Join me as I recap the events from 3000 years ago until now, including quoting John/Cavil's brilliant I'm a machine speach.
Furburger, a gian metal vagina, makes her appearance as does Stumpy, Felix's crusty companion. The revolution is resolved and on the whole I quite liked it. The unlikely return of Lt. Aaron Kelly is most welcome, as is Baltar, and George W. Bush makes an appearance.
my son, apparently, likes cats
I break format and do a six scene breakdown with hillarious pictures and screen-caps. Seriously, you want to check out this fun times full of outrageous type-setting! Oh, also I childe Roslin (again), defend Zarek and use the word microcosm-esque in a sentance.
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Ostatnie opinie
- Wszystkie opinie
- Pozytywne
- Neutralne
- Negatywne
At Glen Mhor Bed and Breakfast, you'll be just a 5-minute drive from Horseshoe Falls and Skylon Tower. Guests looking to stay active can check out the nearby hiking/biking trails, and free perks include WiFi, self parking, and full breakfast daily between 8 AM and 9:30 AM. Other highlights at this Victorian bed & breakfast include a terrace and a garden.


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Students' Frequently Asked Questions
Find answers to questions that students most frequently ask about Macmillan Practice Online. The questions have been divided into categories: just click on a category to open the list of questions and answers
About Macmillan Practice Online
What is in Macmillan Practice Online?
Macmillan Practice Online is an online English course with a Markbook/Gradebook where your results and progress are saved. In addition you will have access to the Macmillan English Dictionary Online and if you are assigned to a class there will be a messaging function to email your teacher.
What are courses?
A Macmillan Practice Online course is an online course that is made up of a collection of resources to practise language, listening, vocabulary and pronunciation including web projects, videos and games. Each course contains up to 200 interactive resources.
What types of course are available?
There are many different types of Macmillan Practice Online courses. These include general, business, academic and exam practice courses as well as courses that accompany Macmillan coursebooks.
Click here for a full list of courses available.
I would like my teacher to view my scores and progress on my Practice Online course, is that possible?
Yes, your teacher can give you a unique password to join their Macmillan Practice Online class. You simply log into your Macmillan Practice Online course click on 'Join a class' and enter this password. Your teacher will then see your course, your scores and your progress. You will also be able to send and receive messages.
Can I do a course without a teacher?
Yes, it is not necessary to work with a teacher on your Macmillan Practice Online course, you can use the course independently.
How long does the course last?
Your Macmillan Practice Online course lasts one year from the date that you activate your code. You can see your course activation and expiry date in the My account area of your Macmillan Practice Online.
How much are the courses?
The courses range in price from £9 – £17. Click here to see a full list.
Is the CPE Practice Online course changing to reflect the new exam?
The CPE exam is changing. From 2013, the exam will have a new format. The Reading and Use of English paper will be combined. We are publishing a new and updated course called Cambridge English: Proficiency Practice Online. If you are already using the CPE course, you will have access to the old course and the new version.
What are the technical requirements for Macmillan Practice Online?
Component Required Cookies Allowed Pop-ups Allowed Operating system Windows >= XP Web browser Chrome 10+
Internet Explorer 8+
Safari 4.1+
Firefox 4.0+


text: I decided to try to move one of my tree stands today and we always use 2-way radios. I let the wife know when I am about to go up a tree and again when I am strapped in tightly. Then again when I get back on tera firma. So, today I also took the chain saw as there was some stuff I had to cut so as to be able to see and shoot. When I got ready to go up, the radio was missing. Search as I might, it stayed hidden. So, back to the house I go on the atv. Wife laughed. I told her to give me time to get back then talk every now and then on the radio until I found it. It worked but made me a bit red faced. Was this a senior moment?


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As the name implies, East Lansing is located directly east of Lansing Michigan. Originally it had been settled as a college town, the very first houses were built right across the street from Michigan State University, which was founded in 1855. But since then, East Lansing has grown to eight times the size of the original town, and continues to grow and prosper!
Parks & Recreation
East Lansing, Michigan is home to more than 20 community and neighborhood parks. The city also includes the Northern Tier Trail: 4.5 miles of paved pathway perfect for hiking, biking, rollerblading, or walking. And for the four-legged family members, East Lansing has the Northern Trail Dog Park: a two-acre, double gated, fenced, off-leash dog park complete with a water station, shade trees, and more!
Arts & Entertainment
Being a college town definitely has it's perks, and everything that Michigan State University has to offer by way of the fine arts is definitely one of them. See a show at the MSU Wharton Center for Performing Arts, or spend a day looking around in the MSU Broad Art Museum!
Annual Festivals
From June until October East Lansing hosts a city-wide Farmers Market in Downtown East Lansing. But the summer here has so much more to offer than just that. Every Saturday night from 7-9 PM there is the Summer Concert Series, there is also the Moonlight Film Series, a free movie festival, and the Summer Solstice Jazz Festival.
As previously mentioned, East Lansing is home to Michigan State University (MSU): a nationally ranked, academic residential college. 50,000 students come in from all 50 states and 130 different countries!
For those who aren't quite old enough to head off to college, the East Lansing Public School District is a premier academic system educating 3,500 students throughout the East Lansing Area.
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Company Report
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NSP Seizes 74 Pounds of Pot in Stop Near North Platte
Two people were arrested near North Platte Friday after a state trooper found 74 pounds of marijuana in their vehicle.
The Nebraska State Patrol says 36-year-old Jared Case, of St. Petersburg, Fla., and 30-year-old Izula Maximillen, of Arcata, Calif., were arrested for marijuana possession and child endangerment. A two-year-old traveling with the pair was placed in protective custody.
A trooper stopped the minivan being driven by Case around 3:30 p.m. Friday near the North Platte exit off Interstate 80 for speeding and not staying in the lane.
The patrol says a drug dog taken to the scene alerted to the presence of drugs. The troopers say they found the pot in an external luggage carrier on top of the minivan.
The duo was taken to the Lincoln County Detention Center on charges of possession of marijuana with intent to deliver and child endangerment.


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IRVING, Texas -- As Tony Romo bears the brunt of another elimination game loss following his three-interception effort against Washington on Sunday, Cowboys coach Jason Garrett defended his starting quarterback.
"We have great belief of Tony Romo as our quarterback, Garrett said. "Tony has won a lot of big games for us, too, to get us to the point where we can play for the division in Week 17 in consecutive years. We all know that we want to take the next step and Tony is going to be a big part of that going forward. I think you have to understand the whole body of work. I think you have to understand that winning is how we get evaluated. He's done a lot of great things for this franchise. We're excited about him being our quarterback."
Romo is signed through 2013 but will count $16.8 million against the salary cap. The Cowboys would like to sign him to an extension to lower that figure and help upgrade the roster around Romo. He put off contract talks at the beginning of the season.
Romo finished with a career-high 4,903 yards with 28 touchdown passes and a career-high tying 19 interceptions. He was sacked 36 times for the second straight year.
Garrett said Romo's pass on the first interception sailed on him to Kevin Ogletree and the second one, a deep ball to Miles Austin, was too far inside. The third turnover, which effectively ended the Cowboys' chances, was a result of not getting enough air on the ball to the flat to DeMarco Murray.
"So you can try and put a lot of different spins on it, but I think what you have to do more than anything else is simply try to analyze what happened on those three plays, and that's the explanation," Garrett said. "And one of the things you can say, 'Were the emotions too big?' I don't think so. Did one snowball into the next? I really don't think so. I thought his demeanor throughout the ball game was good. He settled down. We got going again, and then at that critical moment in the ball game, they made a good play and we didn't make that play. And that was a really important play in the ball game, obviously."


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Ajith and director Vishnuvardhan's untitled flick has Taapsee Pannu playing second fiddle. The makers had confirmed the participation of the Billa jodi Nayan and Ajith, along with beefcake Arya on board. Produced under AM Ratnam's Sri Surya Movies, this one will be a racy thriller with Arya and Ajith delivering the punches.
Leaving the men to work, the movie does have a good dose of glamour too. As Tatoo lady Nayanthara play Ajith's bad girl, Aadukalam actress Taapsee Pannu is delighted to land a role in an Ajith flick. She pairs opposite Arya, after almost disappearing from the scene. The actress did have a lot of skin show going in Tollywood, but no substantial roles at hand post her unimpressive Vanthaan Vendhran release. The new Ajith flick is almost done with the pre-production and the first look posters will be out soon. Music for this release is by Yuvan Shankar Raja, who had his Billa 2 audio album open to a thundering response recently. Bagging another Ajith flick, Yuvan fans are a happy lot. Having glamour girls Nayan and Taapsee sign up for this release, the movie goes on floors at the fag end of May.


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Friday, February 15, 2008
Flowers for you...
Hi y'all! Happy Friday. I am sooooooo ready for a relaxing weekend! How about you? This card I'm sharing today is one I did for the Gina K. Designs Challenges last weekend for her release party. To be honest with you, I'm not all that sure about my card. Colors wrong? Paper wrong? It just doesn't "sing" for me. I know what I was going for! I used my distress pad (Tea Dye) and just rubbed it all over the paper before stamping my image and coloring it with watercolor pencils. I was going for a rustic, almost "dirty"look like this little card escaped and rolled around in the garden. :-) Wierd, I know! The layout was thanks to Erika Martin and it's really unique. I used my Nesties to cut the circle and then just layered it all up. I did sew a zig zag around the circle. Stamped my sentiment and cut it out using a 1" circle punch which I then cut in half and layered under my ribbon. That's it!
Well, I have a list a mile long of things on my to do list! Hope you have a wonderful day! The sun in shining here which is sooooo nice. Hope it is where you are too!
p.s. Edited To Add: Today Nina of the Queen Kat Royal Court dubbed me a "Card Making Princess!"
Do I get a crown??? :-) Very cool!! Thanks Nina!! Wait 'til I tell my family they must now address me as "Princess!" hee hee!
Posted by Julie Koerber at 8:45 AM
Labels: Gina K. Designs
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Working on our first beer can designs with @vailbrewingco - stoked on this project! You can help fund their canning line if you go to indigogo.com and search "VBC" - thanks everyone! #kinddesign #vailbrewery #vail #colorado #madeinCOLORADO...


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Wayne Robinson
Listed by Keller Williams Lakes & Mountains Realty
Single floor living at its best. 3 Bedroom ranch in low tax Wakefield. Perfect for the young growing family or retirees. Low maintenance. Large rooms. 2 Car garage under with double doors into the basement. Separate monitor heater in basement. Pellet stove. Very efficient home. Enjoy all the lakes region has to offer. Near lakes & mountains
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