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Entamoeba histolytica, an amitochondriate protozoan parasite that relies on glycolysis as a key pathway for ATP generation, has developed a unique extended PPi-dependent glycolytic pathway in which ADP-forming acetyl-coenzyme A (CoA) synthetase (ACD; acetate:CoA ligase [ADP-forming]; EC 126.96.36.199) converts acetyl-CoA to acetate to produce additional ATP and recycle CoA. We characterized the recombinant E. histolytica ACD and found that the enzyme is bidirectional, allowing it to potentially play a role in ATP production or in utilization of acetate. In the acetate-forming direction, acetyl-CoA was the preferred substrate and propionyl-CoA was used with lower efficiency. In the acetyl-CoA-forming direction, acetate was the preferred substrate, with a lower efficiency observed with propionate. The enzyme can utilize both ADP/ATP and GDP/GTP in the respective directions of the reaction. ATP and PPi were found to inhibit the acetate-forming direction of the reaction, with 50% inhibitory concentrations of 0.81 ± 0.17 mM (mean ± standard deviation) and 0.75 ± 0.20 mM, respectively, which are both in the range of their physiological concentrations. ATP and PPi displayed mixed inhibition versus each of the three substrates, acetyl-CoA, ADP, and phosphate. This is the first example of regulation of ACD enzymatic activity, and possible roles for this regulation are discussed.
Entamoeba histolytica, a protozoan parasite that causes amoebic dysentery and amoebic liver abscess in humans, is a leading cause of morbidity and mortality due to parasitic disease worldwide, second only to malaria (1). This parasite has a two-stage life cycle, existing as infectious cysts or motile trophozoites that can reside in the anaerobic confines of the human colon and cause disease. After ingestion of cysts and excystation to release trophozoites, asymptomatic infection can occur when trophozoites remain confined to the intestine. However, the parasite can break through the epithelial lining of the intestine and enter the bloodstream, resulting in an invasive infection. Cell lysis, phagocytosis, and trogocytosis all play a role in pathogenicity and contribute sources of nutrients to this metabolically limited parasite (2).
E. histolytica lacks a functional tricarboxylic acid (TCA) cycle and oxidative phosphorylation and consequently relies on substrate-level phosphorylation to provide high-energy compounds. This amitochondriate protozoan utilizes an unusual PPi-dependent glycolytic pathway (Fig. 1) in which ATP-dependent phosphofructokinase is replaced by PPi-dependent phosphofructokinase and pyruvate kinase is replaced by a PPi-dependent pyruvate phosphate dikinase (3–5). The pyruvate end product of glycolysis is converted to acetyl-CoA by pyruvate:ferredoxin oxidoreductase (PFOR) rather than pyruvate dehydrogenase. Acetyl-CoA can then be broken down to ethanol by a bifunctional aldehyde-alcohol dehydrogenase (ADHE) (6) or to acetate by ADP-forming acetyl-CoA synthetase (ACD; acetate:CoA ligase [ADP-forming]; EC 188.8.131.52), as summarized in the following equation: acetyl-CoA + ADP + Pi ⇆ acetate + CoA + ATP.
This extended PPi-dependent glycolytic pathway increases the ATP output per glucose molecule to 3 when ethanol is produced and to 5 when acetate is the product. ACD was first identified with acetate-forming activity in E. histolytica extracts, and the partially purified enzyme was shown to have activity in both directions of the reaction (7). The only other characterized eukaryotic ACD is that from the amitochondriate Giardia lamblia (8–10), for which only substantial acetate-forming activity has been detected (10).
Although not widespread, ACD has been identified in all three domains and is postulated in the conversion of acetyl-CoA to acetate. In archaea, ACD was first identified in Pyrococcus furiosus (11, 12), a thermophilic anaerobe that ferments carbohydrates and peptides, and it has now been identified in a number of other archaea, including the thermophilic anaerobes Pyrococcus woesei, Desulfurococcus amylolyticus, Hyperthermus butylicus, and Thermococcus celer (13, 14), the hyperthermophilic anaerobic sulfate reducer Archaeoglobus fulgidus (15), and the halophilic aerobe Halobacterium saccharovorum (13, 14). Native ACD has been purified from P. furiosus (11, 12) and Haloarcula marismortui (16), and recombinant ACDs have been characterized from P. furiosus (17), A. fulgidus (18), Methanococcus jannaschii (18), H. marismortui (16), Pyrobaculum aerophilum (16), and Thermococcus kodakarensis (19).
A similar role for ACD in acetate production has been proposed in bacteria. Most bacteria have a phosphotransacetylase (PTA) and acetate kinase (ACK), which form a predominant pathway for acetate production from acetyl-CoA. ACD from the phototrophic Chloroflexus aurantiacus has recently been purified, and the recombinant enzyme was characterized (20). The syntrophs Pelotomaculum thermopropionicum (21) and Syntrophus aciditrophicus (22) and the propionic acid-producing Propionibacterium acidipropionici (23) all lack genes for the PTA-ACK pathway and instead have acd genes. ACD has been identified via proteome analysis of P. acidipropionici cells grown under acetate-producing conditions, but the enzyme has not been purified or characterized (23). Likewise, the acetate-producing archaea listed above that have ACD lack genes encoding the PTA-ACK pathway.
ACD is part of a superfamily of nucleoside diphosphate-forming acyl-CoA synthetases, which includes the closely related citric acid cycle enzyme succinyl-CoA synthetase (SCS; EC 188.8.131.52) as well as ATP citrate lyase (EC 18.104.22.168) and malate thiokinase (EC 22.214.171.124) (9). SCS has been extensively studied kinetically and structurally (24–26), and a three-step enzymatic mechanism has been proposed (27), with E in the equations representing the enzyme of interest: (i) succinyl-CoA + Pi ⇆ E∼succinyl-P + CoA; (ii) E∼succinyl-P ⇆ E-His∼P + succinate; (iii) E-His∼P + ADP ⇆ ATP.
Based on their studies with P. furiosus ACD, Brasen et al. (28) proposed that ACD proceeds through a mechanism similar to that of SCS, but with an additional step in which the phosphoryl group is transferred from a His residue on the α-subunit to a His residue on the β-subunit before transfer to ADP in the final step to produce ATP. Here we report the biochemical and kinetic characterizations of the recombinantly produced ACD from the intestinal parasite E. histolytica and reveal its ability to function in either acetate production or utilization in vitro. Our results confirm that both the acetyl-CoA-forming and acetate-forming activities of ACD are present in cell lysates from E. histolytica trophozoites. Inhibition studies suggest E. histolytica ACD (EhACD) is regulated through inhibition by ATP and PPi in the acetate-forming direction.
Materials.Chemicals were purchased from Sigma-Aldrich, VWR International, Gold Biotechnology, Fisher Scientific, Atlanta Biological, Life Technologies, and JR Scientific.
Growth of E. histolytica trophozoites and preparation of cell lysates.E. histolytica strain HM-1:IMSS (kindly provided by Lesly Temesvari, Clemson University) was cultured axenically at 37°C in TYI-S-33 medium (29). Cells were harvested by incubation on ice for 10 min and centrifugation at ∼350 × g for 5 min at 4°C. The pellet was resuspended in 0.8 ml lysis buffer [50 mM Tris-HCl (pH 7.3), 1 mM 4-(2-aminoethyl)benzenesulfonyl fluoride hydrochloride, 0.015 mM E-64, 0.02 mM pepstatin A, 5 mM 1,10-phenanthroline] and disrupted by five cycles of freezing in a dry ice/ethanol bath and thawing at 37°C. The cell lysate was kept on ice, and enzymatic activity was assayed immediately by using the 5,5′-dithio-bis-(2-nitrobenzoic acid) (DTNB) and hydroxamate assays described below. Total protein concentration was determined by using the Bradford method (30) with bovine serum albumin as the standard.
Production and purification of E. histolytica ACD.A codon-optimized E. histolytica acd gene was synthesized by Genscript and cloned into pET21b (Novagen) for production of recombinant E. histolytica ACD in Escherichia coli. The pET21b-ACD plasmid was transformed into E. coli Rosetta2(DE3)pLysS cells (Novagen), and cultures were grown in Luria-Bertani medium (LB) containing 50 μg ml−1 ampicillin and 34 μg ml−1 chloramphenicol, with shaking at 200 rpm at 37°C to an A600 of ∼0.8. Protein production was initiated by the addition of isopropyl β-d-1-thiogalactopyranoside (IPTG) to a final concentration of 0.5 mM. Cultures were grown overnight at ambient temperature and harvested by centrifugation. Cells were suspended in ice-cold buffer A (25 mM Tris-HCl, 150 mM NaCl, 10% glycerol, 20 mM imidazole [pH 7.4]) and disrupted by two passages through a French pressure cell at 138 MPa. The cell lysate was clarified via ultracentrifugation at ∼ 98,000 × g for 1 h at 4°C, and cell extract was applied to a 5-ml HisTrap HP nickel affinity column (GE Healthcare) equilibrated with buffer A. After extensive washing with buffer A to remove unbound protein, the column was developed with a linear gradient of 0 to 0.5 M imidazole in buffer A. Fractions containing active enzyme were pooled, dialyzed overnight in buffer (25 mM Tris-HCl, 10% glycerol [pH 7.0]), aliquoted, and stored at −80°C. The enzyme was judged to be electrophoretically pure by SDS-PAGE analysis. Protein concentrations were determined based on the absorbance at 280 nm using a Take3 microvolume plate (BioTek).
Determination of molecular mass.The native molecular mass of EhACD was determined by gel filtration chromatography through a Superose 12 column (GE Healthcare) preequilibrated with 50 mM Tris-HCl, 150 mM NaCl (pH 7.0). The column was calibrated with cytochrome c (12.4 kDa), carbonic anhydrase (29 kDa), albumin (66 kDa), amylase (200 kDa), apoferritin (443 kDa), and thyroglobulin (669 kDa) and developed at a flow rate of 0.5 ml per minute.
Determination of kinetic parameters for EhACD.Pseudo-first-order reaction kinetic determinations were performed in both directions of the reaction. Enzymatic activity in the acetate-forming direction was determined by measuring the release of CoA with a sulfhydryl group (CoASH) from acyl-CoA by using Ellman's thiol reagent (DTNB) (31). Production of NTB2− by CoASH cleavage of DTNB was measured spectrophotometrically at 412 nm. Reaction mixtures (50 mM Tris-HCl, 0.3 mM DTNB, 6 mM HEPES [used to solubilize DTNB] [pH 7.3]) with varied substrate concentrations were preincubated at 37°C for 5.5 min. Assays were performed in 96-well plates in 0.2-ml reaction volumes. Reactions were initiated by the addition of enzyme, and absorbance was measured every 4 s at 37°C by using a Synergy HT multimode microplate reader (BioTek). Initial velocities were converted to micromoles of CoASH formed by using an extinction coefficient (ε) of 13.6 mM−1 cm−1 for the thiophenolate NTB2− anion. One unit of activity was defined as 1 micromole of product per minute per milligram of protein. In determinations of kinetic parameters for acetate formation, nonvaried substrate concentrations were held at 0.3 mM acetyl-CoA, 8 mM KH2PO4, and 5 mM MgCl2:ADP or 6 mM MgCl2:GDP. For propionate formation, nonvaried substrate concentrations were held at 0.3 mM propionyl-CoA, 12.5 mM KH2PO4, and 3 mM MgCl2:ADP.
Activity in the acetate-forming direction was confirmed using the hexokinase/glucose-6-phosphate dehydrogenase-coupled assay (32). This assay couples ATP formation to the reduction of NADP+ to NADPH, which is measured spectrophotometrically at 340 nm. Reaction mixtures (100 mM Tris-HCl [pH 7.3], 5.5 mM glucose, 1 mM NADP+, 0.2 mM dithiothreitol) with varied concentrations of substrates were preincubated at 37°C for 5.5 min, hexokinase and glucose-6-phosphate dehydrogenase were added, and the reactions were initiated by the addition of ACD. Assays were performed in 96-well plates in 0.2-ml reaction volumes, and absorbance was measured every 4 s at 37°C by using a Synergy HT multimode microplate reader (BioTek). Initial velocities were converted to micromoles of ATP formed by using an extinction coefficient of 6.22 mM−1 cm−1 for NADPH. Concentrations of nonvaried substrates used for both the DTNB assay and coupled enzyme assay were identical.
Enzymatic activity for determination of acetyl-CoA formation was detected using the hydroxamate assay (33, 34). Reaction mixtures (50 mM Tris-HCl [pH 7.5], 300 mM hydroxylamine-HCl [pH 7.0]) with varied substrate concentrations were preincubated at 37°C for 5.5 min, and reactions were initiated by the addition of enzyme. Reactions (0.15-ml reaction volumes) were terminated by the addition of 1 volume of stop solution (2.5% FeCl3, 2 N HCl, 10% trichloroacetic acid), and the absorbance at 540 nm was measured. Product formation was determined by comparison to an acetyl-CoA standard curve. For acyl substrates prepared in ethanol, the final concentration of ethanol in the reaction mixture was kept at 2.5%, which was determined not to affect enzymatic activity. One unit of activity was defined as 1 micromole of acetyl-CoA produced per minute per milligram of protein. In determinations of kinetic parameters for acetyl-CoA formation, the nonvaried substrates were held at 100 mM acetate, 1 mM CoA, 20 mM MgCl2:ATP, and 25 mM MgCl2:GTP. For determination of kinetic parameters for propionyl-CoA formation, the nonvaried substrates were held at the following concentrations: 250 mM propionate, 3 mM CoA, and 20 mM MgCl2:ATP.
Apparent kinetic parameters were determined by varying the concentration of a single substrate while the concentrations of the other two substrates were held constant. For determination of the apparent steady-state kinetic parameters Km, kcat, and kcat/Km, and their standard deviations (SD), nonlinear regression in KaleidaGraph (Synergy Software) was used to fit the data to the Michaelis-Menten equation.
Determination of inhibition parameters.Various metabolic intermediates were tested as effectors of ACD. The IC50s (the effector concentrations that provided half-maximal inhibition) for inhibitors were determined based on Km substrate concentrations by measuring the decrease in enzymatic activity as a function of increasing concentration of inhibitor. Nonlinear regression analysis in Prism 5 (GraphPad Software) was used to fit the data to the log [inhibitor] versus response curve to determine IC50s. To examine the mode of inhibition, enzymatic activity was assayed in each direction in a four-by-four matrix of varied inhibitor concentrations versus varied concentrations of one substrate, with the other two substrate concentrations held constant. Linear regression in KaleidaGraph (Synergy Software) was used to resolve the points of intersection of the inverse plots.
General characterization.The limited characterization of partially purified native E. histolytica ACD reported by Reeves et al. (7) indicated that the enzyme has a narrow nucleotide specificity. To allow a more thorough characterization of the E. histolytica ACD (EhACD), we produced the recombinant enzyme in E. coli. A codon-optimized gene was synthesized by Genscript and cloned into the pET21b E. coli expression vector (Novagen), which encodes the addition of a C-terminal His6 tag to the recombinant protein. The calculated subunit molecular mass of the recombinant His-tagged protein was 78,999 Da. The molecular mass of the purified enzyme was estimated to be ∼150 kDa based on gel filtration chromatography, suggesting that EhACD is dimeric, as found with the G. lamblia ACD (10). The optimum temperature for ACD activity was determined to be 55°C in the acetyl-CoA-forming direction but could not be determined in the acetate-forming direction due to limitations of the equipment used for measurements. This unexpectedly high optimal temperature may have been an artifact from the assay conditions or may represent an unusual stability of this protein, but it was not analyzed further. However, activity was routinely assayed at 37°C, the temperature E. histolytica trophozoites would typically encounter within the human host. The enzyme had approximately 70% activity at this temperature versus its optimum temperature.
The requirement for a divalent cation was examined and ACD was found to have the highest activity with Mg2+ and Mn2+ and substantial activity with Co2+ in both directions of the reaction, but only weak activity with Ni2+, Zn2+, Ca2+, and Cu2+ (Fig. 2). The preference for Mg2+, Mn2+, and Co2+ is similar to that of other ACDs (10, 11, 18).
ACD enzymatic activity was observed in both directions of the reaction, and the reaction rate was linearly dependent on enzyme concentration. Specific activity in the acetate-forming direction (85 ± 1.2 μmol min−1 mg−1) was approximately half that observed in the acetyl-CoA-forming direction (180 ± 2.0 μmol min−1 mg−1). Similarly, measurement of ACD specific activity in E. histolytica cell extracts showed that activity in the acetate-forming direction was approximately half that in the acetyl-CoA-forming direction (0.14 ± 0.022 μmol min−1 mg−1 versus 0.29 ± 0.004 μmol min−1 mg−1). In contrast, ACD activity in Giardia cell extracts could only be detected in the direction of acetate formation (10). Although the purified recombinant Giardia ACD had activity in both directions of the reaction, the acetyl-CoA-forming activity was ∼4% of the acetate-forming activity (9).
EhACD has a limited acyl-CoA substrate range, with the highest activity observed with acetyl-CoA and lower activity observed with propionyl-CoA (16% ± 2.8% activity versus that with acetyl-CoA). Less than 1% activity was observed with isobutyryl-CoA and isovaleryl-CoA at a 0.3 mM final concentration, and no activity was observed with butyryl-CoA, succinyl-CoA, or phenylacetyl-CoA. ADP and GDP gave the highest activity (100% ± 1.0% and 106% ± 3.7% activity, respectively), although activity was also observed with IDP (28% ± 3.8%), CDP (7.0% ± 0.2%), and UDP (2.8% ± 0.1%); no activity was observed with TDP as the substrate. PPi was tested with AMP as the phosphoryl acceptor, but no activity was observed under the conditions tested.
In the acyl-CoA-forming direction, acetate was the favored substrate and propionate and butyrate could also be used, with relative activities of 72% ± 1.5% and 5.6% ± 0.2%, respectively, versus acetate. Less than 4% activity was observed with isobutyrate, valerate, isovalerate, hexanoate, heptanoate, octanoate, succinate, or phenylacetate as the acyl substrate. Activity observed with ATP (representing 100% activity) was nearly double that with GTP (57% ± 0.4%) and substantially higher than the activities observed with ITP (15% ± 0.2%), CTP (4.1% ± 0.3%), or UTP (1.6% ± 0.2%); no activity was observed with TTP or PPi.
Kinetic parameters.Kinetic parameters for EhACD were determined in both the acetate-forming and acetyl-CoA-forming directions (Table 1). Activity decreased at substrate concentrations above 6 mM ADP or 25 mM ATP in the respective directions of the reaction, so the concentrations of these substrates were held below saturation in determinations of kinetic parameters. Under these conditions, acetyl-CoA and Pi in the acetate/ATP-forming direction and acetate and CoA in the acetyl-CoA-forming direction followed Michaelis-Menten-like kinetics. The enzyme displayed similar Km values for acetyl-CoA and propionyl-CoA in the acetate-forming direction of the reaction. However, the turnover rate, kcat, was approximately 5-fold higher with acetyl-CoA versus propionyl-CoA, resulting in ∼4-fold-higher catalytic efficiency with acetyl-CoA. The Km for Pi was similar with acetyl-CoA or propionyl-CoA as the acyl substrate, but the Km for ADP was reduced ∼2-fold with propionyl-CoA versus acetyl-CoA. Similar Km and kcat values were observed with ADP and GDP (Table 1). The Km values observed for EhACD are similar to those reported for the Giardia ACD (0.02 mM, 0.23 mM, and 1.59 mM for acetyl-CoA, ADP, and Pi, respectively) (10).
As the enzyme assay used for determination of kinetic parameters in the acetate-forming direction of the reaction measures the CoA release in the first step, we confirmed our results by using a coupled enzyme assay that measures ATP production in the last step of the reaction. Similar Km and kcat values were observed with this assay (acetyl-CoA, 0.017 ± 0.001 mM [Km] and 130 ± 14 s−1 [kcat]; KH2PO4, 2.1 ± 0.21 mM and 180 ± 16 s−1; ADP, 0.94 ± 0.16 mM and 140 ± 7.2 s−1).
In the acetyl-CoA-forming direction of the reaction, the Km for acetate was ∼2-fold higher than that for propionate (Table 1); however, the kcat values were similar for both substrates, unlike in the opposite direction. Thus, the catalytic efficiency of EhACD with acetate was only 2.6-fold higher than with propionate. The Km for CoA was ∼2-fold elevated with propionate versus acetate as the acyl substrate, but the Km for ATP was slightly reduced. The Km values for ATP and GTP were similar, but the turnover rate was reduced 1.8-fold with GTP. The overall catalytic efficiency with ATP was only 1.5-fold higher than with GTP, indicating that both are suitable substrates. Although weak activity was observed with butyrate, kinetic parameters could not be determined, as the enzyme was unsaturable for butyrate even at concentrations as high as 1 M.
Enzyme inhibition.Since ACD can function to produce either ATP or acetyl-CoA, regulation of this enzyme may be important for maintaining proper levels of these metabolites in the cell. An array of metabolic intermediates was tested as effectors for ACD (Fig. 3). ATP and PPi were found to be potent inhibitors in the acetate-forming direction of the reaction, producing greater than 95% inhibition at a 10 mM final concentration (Fig. 3). Additionally, 50% or less activity was observed in the presence of 10 mM glyoxylate (32% ± 2%), NAD+ (47% ± 3%), or glucose-6-phosphate (50% ± 5%). In the acetyl-CoA-forming direction, only weak to moderate inhibition was observed with any of the compounds tested (Fig. 3). PPi had only a moderate effect in this direction, reducing activity to 64% ± 4% when present at a 10 mM final concentration in the reaction mixture. ATP is a substrate in this direction, and although it does inhibit at higher concentrations, we did not examine this further.
In addition to the intermediates shown in Fig. 3, glyceraldehyde-3-phosphate, phosphoenolpyruvate, and oxaloacetate were tested, but only in the acetyl-CoA-forming direction, because they interfere with the assays used for the acetate-forming direction. None of these compounds resulted in any substantial inhibition in this direction, with 85% ± 0.3%, 85% ± 5.1%, and 90% ± 1.4% activity observed, respectively.
IC50s for ATP and PPi in the acetate-forming direction were determined to be 0.81 ± 0.17 mM for ATP and 0.83 ± 0.27 mM for PPi. The estimated intracellular concentrations of ATP and PPi in E. histolytica trophozoites grown in the presence of glucose are 5 ± 2 mM and 0.45 mM, respectively (35), suggesting that regulation of the acetate-forming activity of ACD by ATP and PPi is biologically relevant. The mode of inhibition by each of these compounds was determined by kinetic analysis in which the concentration of inhibitor was varied versus one substrate and the concentrations of the other two substrates were held constant. These analyses suggested mixed inhibition by ATP (Fig. 4) and PPi (Fig. 5) versus each substrate.
Substrate specificity of EhACD and comparison to other characterized ACDs.Although ACD is not widespread, characterization of P. furiosus ACDs has demonstrated that there are at least two different isoforms (11, 12). This was confirmed by characterization of additional enzymes from A. fulgidus (18) and T. kodakarensis (19). The ACD I isoforms prefer acetate and acetyl-CoA and can typically utilize propionate, butyrate, and propionyl-CoA as well but do not tend to use longer or branched-chain acyl/acyl-CoA derivatives (11, 12, 16, 18, 20). The ACD II isoforms prefer longer acyl/acyl-CoA derivatives, such as phenylacetyl-CoA/phenylacetate and indoleacetate as the substrates (12, 18, 19). Based on our characterization of recombinant EhACD and the previous characterization of the partially purified native EhACD (7), this enzyme belongs to the ACD I class. The Giardia enzyme also appears to belong to this class (10).
All ACDs utilize ADP, but the ability to accept GDP as a substrate varies (11, 12, 16). In contrast to SCS, which has an isozyme specific for each (24), and the G. lamblia ACD, which functions only with ADP (10), EhACD is able to utilize ADP and GDP interchangeably, with similar Km and kcat values for each. However, EhACD may be more likely to utilize ADP in vivo, since the estimated intracellular concentration of ADP is higher than that for GDP (3.3 ± 1.2 mM versus 0.7 mM, respectively [35]). Furthermore, the intracellular concentration for ADP is above the Km value for ADP, whereas the intracellular GDP concentration is below the Km value. Like EhACD, Entamoeba phosphoglycerate kinase can use both ADP and GDP but shows a strong preference for GDP, as judged by the order-of-magnitude difference between the Km values for these substrates (4).
In the acetyl-CoA-forming direction of the reaction, the enzyme does not show a clear preference for ATP or GTP, as the kinetic parameters with each substrate are similar. The fact that the intracellular concentrations of ATP and GTP (5 ± 2 mM and 1.8 mM, respectively [35]) are both below the Km values for these substrates suggests that the acetate/ATP-forming direction of the reaction is favored in the cell, even though the purified enzyme has similar activity in both directions. The higher catalytic efficiency observed with either substrate in the acetate-forming direction versus the acetyl-CoA-forming direction is consistent with this.
A comparison of the characterized ACD I enzymes reveals a great deal of variability. All of the characterized enzymes catalyzed the acetate/ATP-forming reaction, but an acetyl-CoA-forming activity was not detected or was very low for the ACDs from G. lamblia (8–10), P. aerophilum (16), and M. jannaschii (18). A range of Km values has been observed for each substrate in both directions of the reaction (Table 2). Within the archaea, for which six ACD I enzymes have been characterized, the Km for a given substrate in the acetyl-CoA-forming direction showed up to an 11-fold range in values. In the acetate/ATP-forming direction, Km values ranged up to 41-fold for acetyl-CoA. The Km values observed for the single characterized bacterial ACD, from C. aurantiacus, were within the range of values observed with the archaeal enzymes. Likewise, the Km values observed for the G. lamblia enzyme were within or near the ranges of those for the archaeal enzymes.
For EhACD, the Km values for acetyl-CoA and Pi were comparable to those for other enzymes, but the Km for ADP was substantially higher. In the acetyl-CoA-forming direction, the Km value for CoA was slightly above the range observed for other ACDs. However, the Km for acetate was 8-fold higher than the highest Km observed for any other ACD, and the Km for ATP was over 20-fold higher. This may suggest that acetyl-CoA formation is not the preferred direction of the reaction and that E. histolytica may employ this direction only under specific environmental or physiological conditions (see below).
Role of the extended glycolytic pathway.The PPi-dependent extended glycolytic pathway branches after production of acetyl-CoA from pyruvate. In one branch, ACD is responsible for production of acetate and ATP; in the other, acetyl-CoA is metabolized to ethanol by ADHE (6). Montalvo et al. (36) and Reeves et al. (7) showed that ethanol and acetate are produced at approximately a 2:1 ratio under anaerobic conditions versus a 1:2 ratio under aerobic conditions. More recently, Pineda et al. (37) found an ethanol-to-acetate ratio of 3.9:1 was produced in cells growing under standard conditions. Upon exposure to oxygen-saturated medium, however, the flux through acetate increased for up to 90 min. The flux through ethanol decreased but gradually recovered over time. Overall, these results suggest ethanol and acetate are both important end products of glycolysis and that flux through both of these pathways may be necessary to generate sufficient ATP while still providing for NAD+ recycling.
ACD may also play a role in ATP production from amino acid catabolism.In addition to increasing ATP production from glycolysis, the presence of ACD and PFOR allows certain amino acids to be catabolized for ATP production. E. histolytica trophozoites scavenge a number of amino acids. Asparagine, aspartate, and tryptophan are broken down to pyruvate, and methionine, threonine, and homocysteine are broken down to 2-oxobutanoate (38). PFOR converts these products, respectively, to acetyl-CoA and propionyl-CoA (7, 39, 40), which can then serve as the substrates of ACD for ATP production.
Regulation of ACD activity.Although metabolic flux analysis using kinetic modeling suggested that ACD has little regulatory effect over glycolysis (35), its roles in amino acid degradation were not taken into account. On the contrary, one might expect that ACD's position in the extended glycolytic pathway of E. histolytica makes it a prime site for regulation in order to properly maintain ATP and acetyl-CoA concentrations. In fact, we have found that EhACD is regulated by both ATP and PPi in the acetate-forming direction, with IC50s that are within the physiological range based on the estimated intracellular concentrations for each (35). An excess of ATP would indicate that the energy needs of the cell are being met, and thus additional ATP production by ACD would be unnecessary. This would prevent costly depletion of acetyl-CoA, which could instead be used by other pathways.
PPi serves as a phosphoryl donor for two PPi-dependent glycolytic enzymes in E. histolytica (4). A buildup of PPi may signal a backup in glycolysis, which would also warrant inhibition of ACD to prevent depletion of acetyl-CoA. Finally, regulation of ACD by two components of the glycolytic pathway may provide a means for shunting acetyl-CoA to ADHE for ethanol production to regenerate NAD+, thus maintaining a proper balance between these two alternative pathways.
Differences between G. lamblia and E. histolytica allow these microorganisms to occupy diverse habitats.Although E. histolytica and G. lamblia rely on similar extended PPi-dependent glycolytic pathways, the E. histolytica ACD is able to function in both the acetate-forming and acetyl-CoA-forming directions of the reaction, whereas the Giardia enzyme is limited to just the acetate-forming direction (10). This divergence may have arisen due to the different environments these parasites inhabit during infection. Entamoeba trophozoites reside in the mucus layer of the colon, where bacterial microflora produce large amounts of the short-chain fatty acids (SCFA) acetate, propionate, and butyrate (41). Giardia colonizes the small intestine, in which the SCFA concentration is relatively low (41), and thus it is unlikely that ACD is needed in acetate utilization.
The total SCFA concentration in the human colon is approximately 100 to 120 mM, with relative molar ratios of 57:22:21 (acetate:propionate:butyrate) (41), giving approximate concentrations of 57 to 68 mM for acetate, 22 to 26 mM for propionate, and 21 to 25 mM for butyrate. Acetate and propionate concentrations are thus in a range that would suggest ACD's activity in the acetyl-CoA-forming direction may be physiologically relevant and that ACD may switch from acetate/ATP production when glucose is present to SCFA utilization when glucose becomes limiting.
Concluding remarks.Although we have speculated on the roles of ACD in Entamoeba, experimental confirmation is still required. In addition to Entamoeba and Giardia, ACD may also be present in other parasites, including Cryptosporidium muris, Blastocystis hominis, and Plasmodium falciparum. The possible existence of ACD in Plasmodium is of particular interest, not only because of its enormous public health significance but also because the parasites retain fully functional mitochondria. Within the complex life cycle of Plasmodium, the intraerythrocyte stages rely primarily on the incomplete oxidation of blood glucose by glycolysis to produce ATP (42, 43), and thus ACD may play a similar role as in Entamoeba. Whether ACD plays similar roles in other parasites must still be investigated, and results from such studies may provide insights into energy metabolism in these important eukaryotic pathogens.
We gratefully acknowledge Kerry Smith (Clemson University) for helpful discussions on this research and comments on the preparation of the manuscript.
Support for this research was provided by National Science Foundation Award 0920274 (Kerry Smith, PI, and Cheryl Ingram-Smith, co-I), a National Science Foundation Graduate Research Fellowship award to C.J., and Clemson University.
- Received 13 August 2014.
- Accepted 5 October 2014.
- Accepted manuscript posted online 10 October 2014.
- Copyright © 2014, American Society for Microbiology. All Rights Reserved.


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MOORHEAD, Minn. (2/21/15)—Concordia hosted the annual Senior Celebration Day and the five seniors responded by helping the Cobbers finish out the regular season with an 82-80 win over Augsburg.
Jordan Deutsch, Scott Flotterud, Brandon Giese, Ryan Rude and Aaron Zimmer all had a hand in the two-point win over the Auggies which helped Concordia finish in fifth place in the final MIAC standings and earn the No.5 seed for the upcoming conference playoffs.
The Cobbers will enter the postseason with a 15-10 overall record and an 11-9 mark in the MIAC. The 15 overall wins and 11 league victories are the second most since current coach Rich Glas took control of Concordia in 2009.
The game seemed to be well in hand for Concordia for most of the game, but as with the two previous contests, the Cobbers' offense slowed to a halt in the latter stages of the second half. Unlike the losses at Carleton and St. Olaf, CC was able to find a way to get back on track and finish off the victory.
The Cobbers led for most of the first half and built the lead to 16 points with 9:18 left to play in regulation. That's when the second-half jitters started to reappear for CC and the Auggies proceeded to go on an 8-0 run to cut the lead in half. Concordia still had a double-digit lead with under one minute to play.
Augsburg got the margin down to three points on a trio of occasions but each time Giese stepped to the free throw line and made both foul attempt shots to keep the Cobbers with a two-possession lead. Giese went 6-for-6 from the free throw line in the final 32-second of play to seal the win for Concordia.
Giese saved his best game of the season for his last inside Memorial Auditorium. He finished with a season-high 21 points and was 10-for-12 from the free throw line.
The other four seniors all factored in the score sheet as well. Deutsch played valuable minutes in the first half and had one rebound. Flotterud was tough underneath the basket and had four rebounds. Rude had four rebounds and three points in leading the CC offense while Zilmmer gave the Cobber unexpected point production as he came off the bench to go 3-for-5 from the floor and finish with a season-high eight points.
Concordia wound up shooting 51.9% (27-for-52) from the floor and 37.5% (6-for-16) from outside the arc in the game. The Cobbers held a 39-32 edge in rebounding
Besides the seniors, juniors Jordan Bolger, Brett Bergeson and Tom Fraase had the biggest impact on the game for the Cobbers. Bolger matched Giese with 21 points and also added nine rebounds. Bergeson and Fraase traded halves of putting up clutch offensive points. Bergeson was on fire from outside the arc in the opening period. He was 3-for-5 from 3-point range and had 12 first-half ponts. He added one more in the second period. Fraase was held off the score sheet in the first half and then responded with 10 second-half points.
Concordia will head into the postseason for the second time in the past three seasons. The Cobbers will head to Arden Hills to take on No.5 seed Bethel in the MIAC quarterfinals on Wednesday, Jan. 25 at 7 p.m.
Upperclassmen Celebrate Senior Day With Win
|Augsburg (11-14/8-12 MIAC)
|Concordia-M'head (15-10/11-9 MIAC)
MOORHEAD, Minn. (2/21/15)—Concordia hosted the annual Senior Celebration Day and the five seniors responded by helping the Cobbers finish out the regular season with an 82-80 win over Augsburg.

subset: commoncrawl


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Welcome to the Fenton Chess Club website. We are based at the Tollgate Hotel, Ripon Road, Blurton, Stoke-On-Trent, ST3 3BS. We meet on Monday evenings from 8:00 onwards. Whether you are a beginner or an expert you can be sure of a warm welcome. We provide the opportunity to play either competitive chess or just social games. We are affiliated to the North Staffs And District Chess Association, the Staffordshire Chess Association and the English Chess Federation.
Use The Tabs Below To Navigate The Site
Sunday, January 29, 2012
Half Year Grading List
The Half Year Grading List has now been published. As stated in an earlier post, team selection should be based on the gradings at the start of the season NOT the new list. The only exception to this is for ungraded players or players with an estimated grade - these players should now use the grading from the half year list. As far as Fenton is concerned this only affects Derrick Wallace who has been re-graded as 128. Jason and Mike have not played sufficient games to be included in the half year list so will continue to use their estimated grade of 60. You can click here to view all of the Fenton grades in the new list.


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The North River Osprey, smooth running and confident. Seahawk hull meets purpose-driven, open Scout layout to create the ultimate big water fishing platform. Master oceans, estuaries and wind-swept reservoirs with the Seahawk's deep entry and offshore bracket, then fill multiple fish boxes with the precise control and expansive room offered by Scout's open design. Fishing has changed from small waters to big. Osprey is the evolution of superior North River design, materials and construction to keep top anglers solidly on the bite.
2019 21′ North River Osprey
Yamaha F115XB four stroke outboard with hydraulic steering (Also available with Yamaha F150XB)
Single binnacle controls & cables
Command Link single gauge & wire kit
Simrad Go-7 GPS chart plotter/fish finder with map chip
Lowrance Link 6 VHF, stainless antenna
EZ Loader 4,700 lb tandem axle trailer with surge brakes
Seat box slider brackets
Diamond plated anchor guards
Anchor locker/storage box in fore deck
Boarding ladder, welded two step w/vertical welded grab handle
Brackets, welded down rigger
Floors, bolt in .190 smooth aluminum with Matson floor coating
36″ wide console with welded windshield
36″ wide bench seat behind center console with brackrest
• Lifetime limited hull warranty to original purchaser
• H116 – 5086 aluminum bottom
• Rigid longitudinal box beam bottom bracing
• Performance matched reverse chines, lifting and turning strakes
• Offshore motor bracket
• Unparalleled overall performance, strength and finish
• Cowled gunwales for added strength
• Welded NR swim deck trolling motor bracket portside
• Stainless steel bow eye and welded transom eyes
• Stainless steel fasteners throughout
• Rear cowling fish boxes and center pylon receiver
• Trim tab brackets (trim tabs optional)
• Transom tie downs
• Zolatone interior paint with clear coat, one color full side paint (20′ Osprey – one color full side vinyl)
• Polished gunwales
• Matson non-skid surface on bow, step deck and starboard swim deck step
• Center console with aft locking door and inner storage tray
• Hydraulic steering
• Five welded rope cleats, (4) under gunwale
• Durable Aluminum Floors
• Full-width aft storage for battery and outfitting components
• Full-length side trays with rod retainers
• Carpet above side trays only
• Cup holders (4) – side tray adjustable mount
• Four North River sport seats (NR4) on 18″ x 20″ aluminum boxes. (Note: 20′ & 21′ Osprey – Two NR4 sport seat on boxes)
• Step deck with fish box rear (single lid with 70/30 divider in box) and dry storage box forward
• In floor fish box aft of center console with dedicated drain and control valve
• Under floor fuel tank with water separating filter
• Center console panel including:
• Fuel gauge
• Blue Sea 6 switch panel
• 12 volt power point and double USB power socket
• 1250 GPH high capacity bilge pump with auto float switch
• Recessed horn in face of console
• Heavy-duty marine battery and tray with on/off switch
• Navigation / Anchor Lights L.E.D.
• Powered ventilation at transom
• L.E.D. interior accent lighting
• EZ Loader tandem axle galvanized trailer with hydraulic disc brakes, chine load guides, slick bunks and L.E.D. lighting
Sales & Service Advantage
We don't just sell boats. We sell knowhow. From our factory certified Yamaha mechanics to the staff in our parts warehouse, everyone at Port Boat House is dedicated to giving you the expertise and the answers you need to get the best from your boat.
Contact Us
We're always available to help with your boating needs. Come in to see us, or email us at firstname.lastname@example.com Us


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(585) 924-4002
435 Commerce Drive
Victor, NY 14564
Temporary Hours as of 3/16/20:
Monday-Saturday 10-6
Sunday 12-5
Regular Hours:
Mon – Fri: 10am-8:00pm
Sat: 10am – 8:00pm
Sun: 11am – 5:00pm
*Ice Cream is not available at this location
With eight retail location in Western New York, you are never far from Watson's Chocolates. Our manufacturing facility is located in Tonawanda, New York, so our products are delivered weekly (sometimes twice a week!) as fresh as can be. Feel free to call ahead to check on our hours of operation, or visit the individual store pages on Facebook. We welcome your visit.


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Garden Greens Oil Booster -15ml
- Description
- Ingredients
- How to use
- Delivery and returns
Blend our ultra-rich Garden Greens Oil Booster with your favourite cream to instantly upgrade your moisturiser.• Enhance your go-to serum or moisturiser with just a few drops.• Apply solo for deep hydration and a luminous glow.• Mix with our Fruit Smoothie Face Cream to create a lighter, rich lotion.
Crambe Abyssinica Seed Oil. Camellia Oleifera (Tea) Seed Oil. Caprylic/Capric Triglyceride. Oleic/Linoleic/Linolenic Polyglycerides. Cucumis Sativus (Cucumber) Oil. Brassica Oleracea Acephala (Kale) Leaf Extract. Spinacia Oleracea (Spinach) Leaf Extract. Brassica Oleracea Italica (Broccoli) Extract. BHT. Tetrahexyldecyl Ascorbate. Tocopherol. Tocotrienols. Helianthus Annuus (Sunflower) Seed Oil. Cymbopogon Schoenanthus (Lemongrass) Oil. Rosmarinus Officinalis (Rosemary) Leaf Extract. Glycine Soja (Soybean) Oil. Eclipta Prostrata (False Daisy) Extract. Melia Azadirachta (Neem) Leaf Extract. Eclipta Prostrata (False Daisy) Leaf Extract. Melia Azadirachta (Neem) Flower Extract. Corallina Officinalis Extract. Chlorella Vulgaris (Algae) Extract. Coccinia Indica Fruit Extract. Solanum Melongena (Eggplant) Fruit Extract. Moringa Oleifera Seed Oil. Curcuma Longa (Turmeric) Leaf Extract. Ocimum Sanctum (Holy Basil) Leaf Extract. Amber Powder. Elaeis Guineensis (Palm) Oil.
Add a 3-4 drops to your serum or cream. Apply solo to skin for added glow.
We are committed to delivering your order as soon as we can. However, due to current circumstances deliveries may take longer than usual. To-door delivery service may be suspended for specific areas, please refer to the latest news announced by our Third Party Logistic - SF Express.
Standard delivery.
• Free standard delivery on orders over HKD$500
• Order by 6pm for delivery within 3-4 business days.
• HK$30 delivery cost for all orders.
Important delivery information:
• We deliver orders Monday-Saturday.
• Business days are Monday-Friday, excluding public holidays.
• We don't deliver on Sunday or public holidays.
• If you place an order on a weekend or public holiday, we will process your order the
next business day.
• Orders cannot be split and must be delivered to a single address.
• PO Box addresses are not accepted. Our delivery service may not be available in
specified areas, including but not limited to Sha Tau Kok.
• We will send you an order confirmation email and an email once your order has been
• All orders will be delivered by a service requiring a signature.
• We do not offer international deliveries or delivery to Mainland China and Macau. You can update your country browsing preferences here. Please read our Terms & Conditions for further information.
Want to return an item? Get in touch here and we'll be happy to sort it out for you.
This product is currently sold out.


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SUBPRIME MORTGAGE : Simple definition – Subprime – Something that is overvalued Its like.. A bank wants to lend to a person X, but while analyzing X credit rating , bank comes to a conclusion that it's highly probable that X won't return back th.
The pain among smaller lenders has parallels with the subprime mortgage crisis last decade. There isn't a standardized definition of subprime borrowers, though it generally encompasses borrowers.
But a new analysis of the mortgage crisis, which cost millions of families. increased housing demand and prices, a classic definition of a bubble. Smith also pointed to securitization. "Subprime.
Rates For Adjustable Rate Mortgages Are Commonly Tied To The Fed Hike Means Adjustable Rate Mortgages Will Rise and Increase Monthly Payments – Adjustable rate mortgages, or ARMs, are popular among many younger homeowners, because they typically have lower interest rates than the more common 30-year fixed rate mortgage. Many ARMs are called a.
· A subprime mortgage is normally made to borrowers with lower credit ratings and it typically carries a higher interest rate that can increase over time.
Subprime Mortgage Crisis The Subprime Mortgage Crisis is an ongoing real estate crisis and financial crisis triggered by a dramatic rise in mortgage delinquencies and foreclosures. In the United States, the crisis had major adverse consequences for banks and financial markets around the globe.
Subprime mortgage. Subprime refers to higher the risk. These are mortgages that are issued to individuals who are often not qualified. That is, the long term monthly mortgage payment is more than their income. Often, these mortgages are issued on the.
Keywords: Mortgage Finance, Foreclosure Crisis, Subprime Lending, paper, subprime mortgages are defined as those in private-label mortgage-backed.
Subprime is a classification of borrowers with a tarnished or limited credit history . Lenders will use a credit scoring system to determine which loans a borrower may qualify for. Subprime loans.
Adjustable Rate Mortgage Index What Is A 7 Yr Arm Mortgage 3 Reasons I'm Paying My Mortgage Off Early Even Though It Doesn't Make Financial Sense – The mortgage we have is a 7-1 ARM, which means the rate is locked in for seven years. We refinanced into that mortgage two years ago, taking extra cash out of home equity to pay off the last of my.An adjustable-rate mortgage (ARM) is a loan in which the interest rate may change periodically, usually based upon a pre-determined index. The ARM loan may include an initial fixed-rate period that is typically 3 to 10 years.How Arms Work How does Featureless work? | Resurgent Arms, LLC – This video will give a quick overview of what makes a "featureless" AR-15. That will be enough for most people. If you want to geek out a bit on the legal stuff, feel free to read below: (Disclaimer: I'm not a lawyer; this isn't legal advice; I'm not liable for anything that may happen as a result of applying this info
subprime definition: used to describe the practice of lending money, especially to buy a house, to people who may not be able to pay it back: . Learn more.
The subprime mortgage crisis was a result of too much borrowing and flawed financial modeling, largely based on the assumption that home prices only go up. Greed and fraud also played important parts.
While there is no common definition of subprime mortgages in Canada, the proportion of such loans has probably fallen from about 5 percent of the market since the financial crisis, said Benjamin Tal,
the U.S. subprime mortgage boom and bust, contrasts.. 5 Although the definition of a subprime borrower varies from lender to lender, the.
Arm Loan Meaning 5 1 Arm Mortgage Definition – 5 1 Arm Mortgage Definition – We are most-trusted loan refinancing company. With our help you can save your time and money when buying a home or refinancing your mortgage.


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Life in the Club
Here at amFOSS, seniors guide juniors on their way in the club. It is a learning and development partnership between someone with vast experience and someone who wants to learn. Mentoring others is one of the most important parts of being in a community - an experience and a skill which is difficult to gain in a typical classroom scenario. Mentors are responsible for tracking their progress, ensuring that they are doing what is best suited for them.
World of Opportunities
There are umpteen number of opportunities out there in the field of Computer Science, and through amFOSS, one can get the right exposure to these opportunities based on the individual's skillset. Everything from internships, hackathons, conferences, higher study programs and summer school programs - all are right at the tip of your fingers.
GSoC Training
Google Summer of Code is an annual program which provides a platform for students worldwide to contribute to Open Source Projects. Selected students have an opportunity to experience and learn the more practical aspects of software development like reading large code bases, programming using coding standards, using a Version Control System, communicating effectively via IRC & e-mail and following a deadline. At amFOSS, over the last 12 years, over 50+ students have been selected as GSoC scholars, with many selected as GSoC mentors. Students receive expert guidance from experienced developers in the open source world, right from basic email and IRC chat ethics to guidance in writing good proposals.
FOSS Talks
FOSS Talks is a regular club event with the aim of getting students used to public speaking and eliminating stage fear. Each week, 3 students are chosen to speak on a topic they choose, for 7 minutes.
FOSS Nights
amFOSS-bi0s Night is an annual cultural event hosted by the students of amFOSS and Team bi0s. This is an event where members get to show off their non-technical skills. Right from cooking food to making all the necessary arrangements, are all taken care by the members themselves. This is a great opportunity for learning to work as a team. The members orchestrate the cultural events with performances in dancing, singing, playing musical instruments, etc.
FOSS Play
FOSS Play is a regular sports event in which the members go out and play. Physical exercise is an essential component of any student's life and we here at amFOSS make sure we don't miss out on that. Marathons, football, cricket, badminton - you name it, we play it!
Social Commitments
Being part of Amrita Vishwa Vidyapeetham (under the Mata Amritanandamayi Math, an NGO with Special Consultative Status to the UN) we get valuable opportunities to lend hands in noble causes in times of dire adversities. During the Kerala floods (2018) which shook the entire state, Amrita University under the guidance of Mata Amritanandamayi set up a 24x7 Hour helpline for providing victim relief. Our students were able to lend a hand in the noble cause, along with the entire college, running shifts day in and day out. Under the ABC (Amala Bharatam Campaign) program by MAM, our students had the opportunity to serve as part of a Clean Up drive conducted in the nearby area.


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- Join an Event
- Create a fundraiser
- SMH Half Marathon
- HBF Run for a Reason
- City2Surf
- New York Marathon
- Star Ball
- Starlight Day
- Bridge to Brisbane
- Chevron City to Surf
- City-Bay Fun Run
- Gold Coast Marathon
- Melbourne Marathon Festival
- Run Melbourne
- Blackmores Running Festival
- Bowral Classic
- Spring Cycle
- Sun Run & Cole Classic
- The Color Run
- Upcoming Events
- Starlight Christmas
- Starlight Movie Month
- Starlight Five Chefs Dinner
- Great Adventure Challenge
- Get Active Events
New York Marathon
Join Team Starlight in the city that never sleeps – the ever changing, melting pot of cultures and epicentre of the arts – New York, New York.
New York City wears many crowns, and for a good reason. It's a destination that people dream of visiting; a true city slicker's delight. It is also home to one of the best and most iconic road marathons in the world. It's true – just walking through this awe-inspiring city is an incredible experience. Now picture yourself pounding the pavements of this concrete jungle on Team Miracle Babies in the 2018 TCS New York City Marathon.
As you run from Staten Island to Manhattan and cross the Queensboro Bridge, you'll feel yourself digging deeper as you push yourself to reach that finish line in Central Park, millions of people on the sidelines cheering you on.
Not only will you have completed the TCS New York City Marathon, you'll also have raised the vital funds to support Starlight in our mission to brighten the lives of seriously ill children and their families.
Limited spots available. If you're interested, call us on 1300 727 827
Find Out
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All about the Paris Fair Event and Schedule 2015
The Paris Fair event is a multiple international Trade Fairs & Expos that takes place annually and attracts participants from across the world. The event incorporates a trade show, fair and exhibition and presents new services and products that will allow you to improve and embellish your life. On this year the fair will take place in the capital of France, Paris. It is scheduled to run from the 29th of April to 10th May 2015. The event will be held at Paris Porte de Versailles and will be open to the general public.
The opening time will be 10am-7pm. Unique to this year, the opening time on Thursday May 1st will be 10am-9pm. And on Thursday the 8th ,there will be a 'nuit foire de paris concert' a concert that will take place at night. This year, it is predicted that there will be around 595,000 participants. The visitors will include professionals in the fields of housing and environment,bathroom accessories and household appliances, home improvement, gardening and pool, real estate, interior decoration and furniture kitchen equipment, heating air conditioning,audiovisual, water treatment and the list goes on and on and you can rest assured that the professions will be vast and diversified. But as it's said, seeing is believing; you just got to get to the fair and see for yourself.
This is an event that I can promise will be exuberant. The fair will also include retailers and decision makers from the home designs, lifestyle, crafts, fashion accessories, entertainment, wine and gastronomy fields such as ,the well being area, daily living, you name it; it will all be there. You also get the opportunity to have professional advice on the various products being exhibited from the actual fabricators and also the decision makers. All you could ever need brought conveniently under one roof. This is a golden opportunity that you cannot afford to miss.
From Wednesday 29 April 2015 to Sunday 10 May 2015
Porte de Versailles – Exhibition Centre
District : Porte de Versailles – Commerce
Metro : Porte de Versailles – RER : Boulevard Victor
Where to Stay in Paris
There's a massive choice, and prices are generally reasonable considering this is a capital city. We'd advise picking somewhere within easy walking distance of a metro so that you can easily get around all the sights in this itinerary.
Our favourite way to find the best deals on accommodation is to book directly on the hotel website. usually have the best rate available thanks to our collaborators that feeds us with Exclusive and live Promo and deals.
- Le Lapin Blanc, a Hotel just a 5-minute walk from Cluny-La Sorbonne Metro station. he best rate available online. You can modify or cancel your booking without fee until 2 days at noon before your arrival. Get a discount of 10% using promocode WEBACK2019 .
- Des Duc Hotels, a few steps from Chatelet, the Louvre and Notre Dame. Get a discount of 10% using code WEBACK.
- Concortel paris hotel, Reserve directly on website and get 15% reduction compared to other tariffs available online. free breakfast during week-end stays reserved via Hotel Website.
- Hotel Saint honore, Just a short walk from the Champs Elysées and the Madeleine, the hotel Saint Honore is ideally situated for direct access to the Paris airports (CDG, Orly) and the 4 main train stations. BEST RATE GUARANTEE Get 25% off for your 3 nights stay !


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Members' Area
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- Utilities:
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ENAT Keywords
All the pages of the ENAT website are tagged with one or more of the ENAT Keywords.
Results 433 to 444 out of 444.
Project Item
30/10/2007 Happy Tourist
The HAPPY TOURIST project aims to offer equal opportunities to people with disabilities in the tourist sector, either as tourists or as staff, based upon the combination of innovative, modular and cost-effective ICT tools, e-learning concepts and on-the-job-training schemes. 0 comments
30/10/2007 DIADA - Developing and Introducing a New Training Approach in the Tourism Sector.
The project aims to identify and define requirements of the demand and supply sides in the tourism sector (both disabled people and tourist industry workers); to prepare an information model for disabled tourists, (by including disabled people in the whole process), and to deliver suitable training programmes for the tourism sector. 0 comments
17/07/2007 Travelling, Volunteering and Learning Activities Generating Employment for the over 55s
The project aims at offering travelling, volunteering, learning, intercultural and income generation opportunities to people over 55 years old from 10 countries. 0 comments
ENAT Library Item
30/10/2007 EC Regulation on Access to Air Transport for People with Reduced Mobility
EC Regulation 1107 of 5 July 2006 specifies the conditions which operators in EU countries must meet for ensuring access to air travel by people with reduced mobility.
30/10/2007 Guide for Passenger Terminal Accessibility
Guidelines by the Canadian Transportation Agency to help air, rail and ferry terminal operators implement the provisions of the Canadian Code of Practice on Passenger Terminal Accessibility, to make their operations more accessible for persons with disabilities.
04/10/2007 Report of the ENAT International Workshop, Athens, October 2006
Workshop Report including summary presentations and speeches by plenary speakers, workshop conclusions and photos from the optional excursion to accessible attractions in Athens.
17/07/2007 Presentation of ENAT in Greek, July 2007
A slide presentation of ENAT, given by the Coordinator, Ivor Ambrose, at a National Conference on Tourism for All, organised by the Hellenic Ministry of Tourism, 12 July 2007.
01/09/2006 ENAT Inaugural General Meeting Presentations
View and download the presentations by plenary speakers at the ENAT Inaugural General Meeting which took place in Brussels, Belgium on the 11th of May, 2006.
News Item
27/09/2007 ENAT Congress Draft Programme published
The Draft Programme of the ENAT International Congress on Tourism for All is now published. 0 comments
19/09/2007 The Scandic hotel chain receives two 'STIL' awards for its accessibility work
STIL, the Stockholm Cooperative for Independent Living, announced the winner of its fourth annual Ace of Hearts award. For the first time ever, the accolade was given to a company and to that company's Disability Coordinator who, with initiative and creativity, has improved accessibility for disabled people. 0 comments
30/07/2007 ENAT Newsletter Number 1, May 2007 and Number 2, July 2007
The Second ENAT Newsletter is now available. 0 comments
18/07/2007 "Tourism for All" is Greek Tourism Campaign Slogan 2007
This summer, "Tourism for All" is the Greek National Tourist Organisation's domestic marketing message, with posters, decorated buses and a TV-spot contributing to the campaign. 2 comments


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Seeing Washington by Trolley. How to Reach by Trolley Points of Interest in the National Capital. Compliments of The Capital Traction Co. Washington Ry. & Elect. Co.
New York. Rand McNally. 1924.
Red white and blue map 12 x 18 inches, folding as issued to pamphlet 9 x 4 inches. 4 1/2 inch split to one fold end, shorter splits (less than 1 inch) to two other fold ends; clean condition. Text on verso explains that the pamphlet "has been designed and arranged to assist the stranger who is seeking to get about Washignton by electric street car", and provides information on over fifty local attractions. Trolley lines and bus lines are shown in red. Prominent on the map are large areas of land reserved for the National Training School for Boys (a juvenile detention center), the U.S. Insane Asylum, and the U.S. Soldiers' Home. Striking color pictorial cover of Uncle Sam and the Capitol. Item #37766
Price: $110.00


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The following statements were posted to the verified Twitter accounts of U.S. President Donald Trump, @realDonaldTrump and @POTUS.
The opinions expressed are his own. Reuters has not edited the statements or confirmed their accuracy.
@realDonaldTrump :
– Even Crazy Jim Acosta of Fake News CNN agrees: "Trump World and WH sources dancing in end zone: Trump wins again…Schumer and Dems caved…gambled and lost." Thank you for your honesty Jim! [0631 EST]
– In one of the biggest stories in a long time, the FBI now says it is missing five months worth of lovers Strzok-Page texts, perhaps 50,000, and all in prime time. Wow! [0655 EST]
– Nobody knows for sure that the Republicans & Democrats will be able to reach a deal on DACA by February 8, but everyone will be trying….with a big additional focus put on Military Strength and Border Security. The Dems have just learned that a Shutdown is not the answer! [0834 EST]
– Thank you to General John Kelly, who is doing a fantastic job, and all of the Staff and others in the White House, for a job well done. Long hours and Fake reporting makes your job more difficult, but it is always great to WIN, and few have won more than us! [0916 EST]
– Where are the 50,000 important text messages between FBI lovers Lisa Page and Peter Strzok? Blaming Samsung![2254 EST]
– Cryin' Chuck Schumer fully understands, especially after his humiliating defeat, that if there is no Wall, there is no DACA. We must have safety and security, together with a strong Military, for our great people![2307 EST]
— Source link: (http://bit.ly/2jBh4LU) (http://bit.ly/2jpEXYR)
(Compiled by Bengaluru bureau)


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Eviction and Ejectment - Criminal Defense - Civil Defense
Law Office of Mark D. Copoulos
Mark Copoulos has over a decade of trial experience litigating evictions, ejectment, criminal defense and real estate matters in the Philadelphia Municipal and Court of Common Pleas. Since 2009, Copoulos has been steadfastly dedicated to winning his clients cases in criminal and civil defense matters. Our firm prides itself on responsive representation at reasonable prices during these difficult economic times. In addition, when you retain our firm Copoulos guarantees that he will handle your case personally. Your matter will not be delegated to an associate or paralegal. Instead when retain our firm you are guaranteed to receive elite representation from a veteran of the Philadelphia courts whose reputation has garnered hundreds of positive reviews on various social media platforms. If you need relentless, devoted, and committed representation please contact our firm. Mark Copoulos will not rest until he has successfully resolved your case.
Mark Copoulos, Esquire
Mark Copoulos is a "Top 40 Under 40" and "Top 100 Criminal Defense Lawyer" according to the National Trial Lawyers Association. He has a perfect 10.0 score on Avvo.com and points with pride to hundreds of positive reviews on various platforms. These reviews matter because they are his "People Score" and indicative of total commitment to the cases he takes on and client satisfaction. Unlike many fly by night attorneys who take the money and do as little work as possible, Copoulos has over ten (10) years of experience working for the client in the Philadelphia Courts to achieve the best possible outcome. His reputation for commitment and performance separates him from other lawyers who may charge reasonable rates but fail to achieve the desired outcome. Since 2012, Copoulos has won criminal defense, evictions, ejectment, tenant defense, landlord-tenant, and real estate cases for people who have chosen his services. Put his experience and devotion to work for you. Contact Mark Copoulos for assistance in efficiently navigating and resolving your civil, criminal or landlord tenant matter.
Practice Areas
Philly's Best Eviction, Ejectment and Criminal Lawyers
Total Commitment to Your Case
Thousands of Satisfied Clients Over Ten Years
Best defense attorney in the city, and most certainly worth the money. We went to trial and hit a home run with a verdict of not guilty. Mark is precise, attentive to detail and superb. He preformed brilliantly, and bestowed within me a new confidence. Even more, he's affordable and fair.
Jeremy M
For the first time ever, I needed the services of a lawyer. I looked at several different lawyers and their reviews in the Philadelphia area, trying to select someone who could help me. When I came to Mark Copoulos, I stopped. I am so glad I did.
Mark is aggressive, and detail oriented, with sharp wit and class. He took his responsibility to us very seriously, but his personality was so cool, that he was actually fun to work with. There are no guarantees in legal battles, but I really felt great about the possibilities with him as our attorney. Oh…and WE WON!
My son Charlie was arrested for drug possession. Somebody recommended that I call Mark. I was worried he was going to be more expensive than I could afford. Mark was great. He didn't even make me pay the entire fee when we settled the case early.
Mark fought hard for me when the Public Defender ignored my case. The judge also seemed against me. He got a continuance and kept fighting until we got the result I wanted. He didn't charge me extra for the extra court appearances and was there to protect me at a time when it seemed like no one else in the world was interested in justice. Thank you Mark!
Criminal Defense Client
Mark represented me in my DUI case in Philadelphia. He is an excellent lawyer, and fought extremely hard to get the results I deserved. I would recommend him to anyone looking for a personal and affordable attorney.
DUI-DWI Client


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Adding structure and focus to your valuable training time is the best way to achieving your goals. Training with a group can offer many more benefits than traditional 1:1 coaching. Whilst it is true that some athletes need that approach to balance the demands of a full-on race season. For almost every other cycling ambition we believe that Group Coaching offers a more versatile and more motivational way to train.
Employing a coach is a big decision; it can be expensive and very demanding. Usually a cyclist or triathlete might only employ a coach to support a racing career, or in preparation for a big event. What if you have broader goals, or similar goals to your friends?
We offer 3 group-coaching options, called 6 hour +, 10 hour + and Multisport. These are fluid and you may join one or another during the year depending on what your schedule might be. They are all additional to an unlimited membership as the coaching plans utilize our fitness classes, and we like to speak to you face to face as often as we can.
• Monthly Training Plan to progressively build your fitness.
• TrainingPeaks Basic account to track fitness metrics and view schedule.
• Mapdec Group Coaching app for guidance, workout tips, feedback and communications.
• 1 Thursday eve per month event to get face to face feedback, consult with your coach and skills workshops.
6 hours of training and riding time a week is the benchmark for the time-crunched cyclist, the off-season enthusiast or the recovering athlete.
This group will have an emphasis on general fitness, skills and wellbeing. You might join this group to start your fitness journey, or to be effective as possible with your time.
You can always do more, and in the group we will guide you though balancing your training hours and intensity.
Membership + £25 pm6 Hour Group Sign Up
10 hours of training is much more of a commitment, and not something to be rushed into. You might commit to this plan all year, or ramp your training up prior to an event or for a season of racing.
The 10-hour plan will generally take a polarised approach that balances workouts of high intensity with workouts of low intensity ones. We also include more strength and mobility workouts that will help you maintain this volume of training safely.
Membership + £35 pm10 Hour Group Sign Up
This group uses a 10 hour per week plan to balance the demands of two or more sports. This is most suitable for athletes who want to be competitive across Run, Bike & Swim events. If you just enjoy the odd run or swim, the 6 or 10 hour plans above may suit you better.
On occasion this group may use an alternative location for the Thursday evening sessions.
Membership + £35 pmMultisport Group Sign Up
On top of these Group Coaching plans we offer 2 more solutions to athletes that need either more flexibility or more focus.
So ask them! If you would like a call back, please leave your number.
We are always available for a quick chat after a fitness class, or drop in to the studio in the daytime for a chat.


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Posted By Ravi Sinha | On 09th, Aug. 2012 Under Video News
Arkane Studios' Dishonored already has a major ace in it's hole with a steampunk assassin and colonial setting, but if that hasn't sold you yet, this new Creative Kills trailer definitely will.
Honestly, it's great to see this much variety within an action game, forget that of the stealth genre. Slow time and possess an enemy to kill with him his own projectile, or just pluck a cannon out of the air and heave it right back.
The best combination has to be swarm of rats however. Combine with Spring Razor and watch the good times roll.
Source: AllGamesBeta
Contact Us About Us Staff Advertise With Us Privacy Policy Terms of Use Reviews Policy Hooked Gamers Community Policy


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Tips for Market Theater
Market Theater Tips
There is no more free zone, but public transportation is easy.
Food & Cultural Tour of Pike Place Market
Use public transportation. The Market is in the free zone.
Parking in the area sucks. Arrive early enough to search for the best parking rate
Comfortable shoes, and warm clothes during the winter months.
Dress for rain, most of the tour is outside.


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HTC girls find second win on the road
WAYLAND – Junior Emily Box led the Holy Trinity girls basketball team to their second win of the season in a 49-46 win over WACO Friday night in Wayland.
Holy Trinity, which will play it's first five games on the road to start the season, held a slim one-point advantage at the end of the first quarter before building a six-point lead at the end of the first half. Junior Taylor Boeding led the way with nine points in the half, including two 3-pointers.
The Lady Crusaders extended the lead to eight points at the end of the third period, but WACO closed the gap to 34-30 with about three minutes to go. In the fourth quarter HTC kept WACO down by as much as six, but a couple of last minute 3-pointers by Kristen Conelly pulled the Warriors within three during the last minute.
Box finished with 11 points and Boeding had 10 leading the scoring for Holy Trinity. Senior Brett Schneider fired in eight points, seven in the second half. Sophomore Ashlynn Haas and senior Madison Mohrfeld each chipped in with four points and junior Eryn Anderson scored three points for the Crusaders.
Holy Trinity takes its 2-1 record to Mediapolis today with the freshman, junior varsity and varsity boys squads with games tipping off at 4:30 p.m.


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John Harold "Johnny" Sherf (April 8, 1913, Calumet, Michigan – August 19, 1991, Dearborn, Michigan) was a National Hockey League player and the first American citizen to have his name engraved on the Stanley Cup.
John Sherf was born and spent his childhood years in Calumet in the state's Upper Peninsula. He participated in youth ice hockey, which was popular in the cold climate of the area. By the time he graduated from Calumet High School his hockey skills were widely recognized. In the working class area where he was raised, he was destined to go to work in the copper mines in the area, continuing his hockey career in the local semi-professional leagues. A local physician took an interest in Sherf and provided him with a scholarship to attend the University of Michigan in Ann Arbor.
Sherf attended Michigan from 1932 until 1936, majoring in history and playing hockey on the varsity team during all four years. He played center at U of M for three seasons. John was the leading scorer in the 1934–35 season with 23 goals and 14 assists in 16 games. In 1935–36, he was the leading scorer of the Western Conference with 33 goals and 10 assists in 17 games. That year, Michigan's opponents scored a combined total of only 30 goals. During his senior year Sherf was elected captain of the hockey team.
Sherf graduated from Michigan in June, 1936. Before he was awarded his degree at the annual commencement ceremony, he was signed to a contract by the Detroit Red Wings of the National Hockey League (NHL). He played several games at the end of the 1935–36 season with the Red Wings top farm team, the Detroit Olympics. He was also called up to play with the Red Wings during the Stanley Cup playoffs that season. The 1936–37 season was Sherf's most memorable as a professional. He played with the American Hockey League's Pittsburgh Hornets, after the Detroit Olympics were moved to Pittsburgh and re-named. At the end of the 1936–37 season Sherf was called up by the Red Wings for the playoffs to help defend their Stanley Cup championship. He collected one point in five games as the Red Wings won their second straight Stanley Cup by beating the New York Rangers three games to two. He was the first American citizen to be a member of an NHL playoff championship team, having his name engraved on the Stanley Cup as a member of the champion Detroit Red Wings.
Johnny Sherf continued his professional hockey career until 1944. He was a member of the Detroit Red Wings for his entire career but was often assigned to the top Red Wings farm team, the Pittsburgh Hornets. Sherf gained recognition both with the Red Wings as well as the Hornets. Sherf is still the seventh leading scorer in the history of the Pittsburgh Hornets, scoring 94 goals and 121 assists in 301 games played. After being called for military service in 1943, he returned to the Red Wings in 1944 before retiring from professional hockey.
He played in 19 NHL regular season games but scored no points. He scored one assist in eight playoff games.
External links[edit | edit source]
|This page uses content from Wikipedia. The original article was at John Sherf. The list of authors can be seen in the page history. As with Ice Hockey Wiki, the text of Wikipedia is available under the Creative Commons Attribution-Share Alike License 3.0 (Unported) (CC-BY-SA).


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Shade Tree Advisory Committee
The Shade Tree Advisory Committee consists of seven members appointed by the Lawrence Township Council. All of the members serve as volunteers.
Our Mission
We advise Planning & Zoning Boards on applications for site plan and subdivisions regarding standards, types and planting specifications for shade trees. We recommend to Township Council programs of shade tree improvement, planting and replacement along public rights-of-way. The Shade Tree Advisory Committee has marked the Township's 19th year as a Tree City with a brochure highlighting the benefits derived from trees and the memorial tree program. See our latest brochure. For more information on how to care for trees see the attached.
We all need trees because of the many benefits. Most agree that trees have an aesthetic appeal and that they also provide shade, wood for construction and wood for fire places. However, in our busy lives we tend to take the many other advantages of trees for granted. Click here to read more information about "Trees - Who Needs Them".
- Arbor Day Celebration
- Memorial Tree Program
- Tree City USA
Members & Liaisons
Current Members:
- David Bosted (Through 12/31/20)
- Edward Brzoza, Alternate #1 (Through 12/31/21)
- Sheila Grant (Through 12/31/19)
- Pam Mount (Through 12/31/21)
- Edward S. Sproles (Through 12/31/20)
- Vacancy (1)
Current Liaisons:
- Councilman Michael S. Powers, Township Council Liaison
- Jim Kownacki, Planning Board Liaison
- TBD, Township Staff Liaison
Check Meeting Agendas and Minutes
Generally, the Committee meets on the 4th Monday of each month at 7:00 P.M. in the Community Development Conference Room in the Municipal Building.
Projects & Activities
The Shade Tree Adivsory Committee regularly participates in the Lawrence Township annual Community Day in October. The photo pictures the Shade Tree Advisory Committee's past display with members drawing the winner's name for a bonsai tree. Pictured left to right are Councilman Michael S. Powers, Shade Tree Advisory Committee member Carmine Di Sanzo, Committee member Janet Kane, and Committee member Edward S.Sproles.


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Charlie Moores recent account of emerald dove on the 10000 birds website reminded me of my trip to Singapore in 2007. I didn't manage to see emerald dove on my visit, but I did see plenty of these - pink-necked green pigeon. Very common but delightfully pretty. Beats the UK's wood pigeon hands down in my book (two of which are in my garden now, vacuuming all the bird seed off the bird table!)
The Davison and Fook (oo-er!) photographic guide to birds of peninsular Malaysia and Singapore describes PNGP as "chubby" and "enormously common". I like that.
The bird featured here was feeding in our hotel garden one afternoon, in city centre grounds that also attracted blue-crowned hanging parrot, asian koel, common iora and plain-throated sunbirds. Nice.


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Web Analytics
Web analytics, as the name suggests, enables one to analyze and also manipulate the properties of the web in a manner so that your business website reaps the optimum of the benefits, as the results of the analysis, if conducted well, are usually far from being uncertain
Why do we need web analytics?
Effective analytical skills are highly essential in case you want to be sure of the pluses and minuses by all means before deciding upon anything. The same applies when you wish to compete with your rivals on the web.
A businessman and his team need to be sharp enough at web analytics before formulating any of the business strategies. It would simply help avoid any confusion, thus making all the thoughts, ideas, and the resulting strategies more lucid.
In spite of all the efforts to make website more effective (including search engine optimization, developing links, making use of the AdWord tool, resorting to Pay Per Click and e-mails) web analytics is still highly crucial. How else would you be getting to know what is it actually that draws traffic to your business website?
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Attract Talent
Have a voice in the Chamber's efforts to promote Helena as a great place to live, work and play.
As a participating Helena WINS employer, help target workers in specific demand occupations to consider Helena's opportunities.
Develop Talent
Work collaboratively with employers who have similar needs to attract specific training opportunities for your employees.
Showcase your business and employment opportunities with emerging workers who will be entering the workforce in the coming weeks, months and years.
Learn how to build advancement opportunities among your current workforce to grow employees' skill sets.
Retain Talent
Be on the cutting edge of implementing employment practices that foster loyalty among your workforce.
Collaborate with other businesses to strategize community services that matter to your workforce.
1. Why should businesses participate in Helena WINS?
With record low unemployment and an aging workforce, it's never been more challenging to attract, develop, and retain talent. Helena WINS brings together businesses, workforce, and resource providers to facilitate new avenues for workforce development. If finding, training, and retaining employees is a challenge for your business, Helena WINS can help.
2. What does it cost?
There are no defined fee structures associated with Helena WINS. Because this is an employee-driven initiative, participating businesses determine how much time they would like to devote to work-based learning programs and opportunities. If additional resources are necessary, it's a business decision to invest in those specific strategies.
3. What size businesses are ideal for Helena WINS?
Businesses of ALL sizes are welcomed to participate in Helena WINS.


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Black Brand apparel
Headed up by ex-HotBike Magazine and ex-S&S man Howard Kelly, Black Brand was launched last year to fill a gap in the mid-range leather apparel market and is distributed exclusively by Biker's Choice.
Of Black Brand, Howard says: "We are a group that is not for everyone. Black Brand doesn't care what kind of bike you ride, just that you ride. Our gear is not what you buy when you want what everyone else has—you need to be the wolf, not the sheep to ride in Black Brand.
"We are making our mark on the cruiser apparel industry, and we are determined to do things that other companies are not. We refuse to be all things to everyone, and we produce apparel for people who want to be different.
"We are the premiere head-to-toe apparel line for cruiser riders, giving you choices, both in styling and function. We will set the direction and trends in the market in such a way that other apparel companies will start to copy our designs. We won't stray from being a motorcycle clothing company, which means we are committed to bringing you the best apparel for the best price, because nobody else has the balls to.
"We started by looking at the apparel lines in the already highly competitive motorcycle industry. We looked at apparel in the lower-end market. We looked at apparel in the high-end market, and we found there was nothing for the mid-line market. That is where we come in.
"We began by analyzing every facet of apparel that was currently available for motorcycle riders. Other manufacturers seemed to be making jackets without considering what a rider, or passenger, might want or need: vents on jackets that were supposed to allow air flow, but were blocked by pockets; women's jackets that rode up while riding, forcing them to buy a larger size just to be comfortable; seams that were added for no apparent reason.
"We made it our mission to make a product that was better. The result?
All of our women's jackets have a hip zipper that allows give in the jacket and prevents it from riding up. Jacket liners that snap back into place instead of a button and fabric loop. A riveted leather strap in all the jackets for easy hanging. An easy way to identify a man's jacket from a woman's, and the right amount of pockets to make jackets useful.
"We listened to what our dealers and their customers had to say and made our apparel better than anything else on the market."
The multi-pocket 'Tempest' two-piece rain suit from Black Brand has a two-year manufacturer's warranty against defects and features a dual storm flap front with an expandable hood and elastic closures at the wrist and ankle. The front and back of the suit has reflective stripes; the pants feature heat-resistant pads on both inner legs and the jacket has hook-and-loop fit adjustments.
The 'Compression' jacket is made from top grain leather, with an asymmetric leather racing stripe overlay. Features include YKK zippers, including zippered sleeves, reflective trim, snap-fit quilted, insulated, removable liner, plus a six-year manufacturer's warranty against defects.


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Ovns JC01 Pod Starter Kit
Sold Out
Ovns JC01 Pod Kit
The OVNS JC01 Pod System is a slim and sleek vape, featuring a lightweight and compact design, draw-activated firing, and three different pods for alternative vaping methods and materials. The OVNS is very thin and small in shape, holding a J**l pod at the very top, above the LED indicator lights. The draw activated firing means that the device can be designed without a need for buttons and can be used in a convenient on-the-go manner. The OVNS JC01 comes with three different pods and coil options for various juice options and nicotine levels.
OVNS JC01 Pod Starter Kit Features:
- Dimensions - 66.5mm by 40mm by 8mm
- All-in-One Design
- 0.7mL Juice Capacity
- Integrated 400mAh Rechargeable Battery
- Ceramic Coil
- Coil Resistances For Pods: 2.0ohm/1.5ohm
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- Draw Activated Firing
- Lightweight and Compact Design
- Replaceable and Refillable Cartridge
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- 1 OVNS JC01 Pod System
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- 1 Instructional Manual
Ovns JC01 Pod Starter Kit has a rating of 5.0 stars based on 13 reviews.
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