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GA (glucoamylase) hydrolyses starch and polysaccharides to β-D-glucose. RoGA (Rhizopus oryzae GA) consists of two functional domains, an N-terminal SBD (starch-binding domain) and a C-terminal catalytic domain, which are connected by an O-glycosylated linker. In the present study, the crystal structures of the SBD from RoGA (RoGACBM21) and the complexes with β-cyclodextrin (SBD–βCD) and maltoheptaose (SBD–G7) were determined. Two carbohydrate binding sites, I (Trp47) and II (Tyr32), were resolved and their binding was co-operative. Besides the hydrophobic interaction, two unique polyN loops comprising consecutive asparagine residues also participate in the sugar binding. A conformational change in Tyr32 was observed between unliganded and liganded SBDs. To elucidate the mechanism of polysaccharide binding, a number of mutants were constructed and characterized by a quantitative binding isotherm and Scatchard analysis. A possible binding path for long-chain polysaccharides in RoGACBM21 was proposed.
- carbohydrate-binding module
- β-cyclodextrin
- maltoheptaose
- Rhizopus oryzae glucoamylase
- starch-binding domain
Starch, the primary source of stored energy in plants, is composed of amylose and amylopectin. The former consists almost entirely of α-(1,4)-D-glucopyranose units; however, a few α-1,6 branches and linked phosphate groups may be found [1,2]. The latter is composed of α-(1,4)-D-glucose segments connected by approx. 5% α-1,6 branching sites [3]. Both amylose and amylopectin fold into helical structures [4,5] and are further organized into the semicrystalline granular form. In the crystalline state, amylose forms double helices [6], whereas in solution it is a flexible chain that can adopt a locally helical shape. In animals, glycogen, similar to the starch in plants, is the storage form of glucose. Glycogen has structural similarities to amylopectin, except that the number of branching sites and glucose units are, in general, more than that of amylopectin. GA (glucoamylase), also known as amyloglucosidase or γ-amylase (EC 18.104.22.168), is a biocatalyst capable of hydrolysing α-1,4 and α-1,6 glycosidic linkages in raw starches and related oligosaccharides to produce β-D-glucose [7,8]. The GA from Rhizopus oryzae (RoGA) comprises a C-terminal catalytic domain, classified as a member of the GH15 (glycoside hydrolase family 15) [9], connected to an N-terminal SBD (starch-binding domain) by an O-glycosylated linker [10]. CAZY (http://www.cazy.org/) classification, which is based on primary sequence similarity, groups the SBD of RoGA into CBMs (carbohydrate-binding modules) family 21 (RoGACBM21) [11,12].
To date, 51 CBM families have been classified, and eight of them (families 20, 21, 25, 26, 34, 41, 45 and 48) have starch-binding activity [13]. The dissociation constants, Kd, for the binding of SBDs to starch are in the micromolar range [14,15]; however, SBDs associated with GA only appear in two families (CBM20 and CBM21). Previous reports on structure-based molecular modelling and NMR spectroscopy of RoGACBM21 revealed that although these two families share very low sequence identity, they indeed have similar biological function and structural folding [16]. Therefore the most recent evolutionary study on CBM20 and CBM21 proposes that these two SBD families are grouped into a new CBM clan [17]. However, the detailed binding mechanisms between CBM21 and starch/glycogen remain unclear.
SBDs can be functionally independent of the catalytic domains and have been proposed to increase the hydrolysis of granular starch by disrupting its structure and concentrating the catalytic domains on the surface of starch [14,18–20]. Ligand binding of RoGACBM21 with starch and soluble oligosaccharides was determined by Chou et al. [15]. The Kd values were of a similar order of magnitude to that of CBM20 from AnGA (Aspergillus niger GA), but the maximal amount of bound protein (Bmax) of RoGACBM21 was 40- to 70-fold higher than that of AnGACBM20 [21,22]. The binding affinity between RoGACBM21 and soluble ligands, βCD (β-cyclodextrin) and G7 (maltoheptaose), was measured as approx. 5 μM [15].
Three-dimensional structures of several starch-binding CBM superfamily members have been reported: CBM20 from AnGA [23], CBM21 from RoGA [16] and Homo sapien phosphatase 1 (PDB: 2EEF), CBM25 and CBM26 from Bacillus halodurans maltohexaose-forming amylase [24], CBM34 from Thermoactinomyces vulgaris α-amylase [25], CBM41 from Klebsiella aerogenes pullulanase [26] and CBM48 from Rattus norvegicus AMP-activated protein kinase [27]. In the present study, we determined the crystal structures of apo-SBD and the complexes with βCD (SBD–βCD) and maltoheptaose (SBD–G7). Additionally, a number of RoGACBM21 derivatives containing point mutations in aromatic residues (Y32A, W47A, F58A, Y67A, Y83A, Y93A and Y94A) and hydrophilic residues (N29A, K34A, N50A, E68A, N96A and N101A) were isolated and studied. We describe detailed interactions between RoGACBM21 and two substrates (βCD and G7) and propose a polysaccharide-binding path.
Protein expression and purification
Recombinant enzyme expression, purification and the functional assay for RoGACBM21 have been reported previously [15]. However, the quality and quantity of full-length RoGA protein expression was poor due to the flexible O-glycosylated linker. Briefly, the DNA fragment encoding RoGACBM21 was cloned into the pET23a(+) expression vector and overexpressed in Escherichia coli BL21-Gold (DE3) cells (Novagen). The recombinant SBD sample was purified by His-BindR affinity column chromatography (Novagen). The purified RoGACBM21 sample was dialysed against sodium acetate buffer (50 mM, pH 5.5). The resulting C-terminally His6-tagged SBD (11.65 kDa) was purified by Ni-NTA (Ni2+-nitrilotriacetate) affinity chromatography with a final yield of 5 mg of purified protein per litre of cells.
Site-directed mutagenesis
All RoGACBM21 mutants were generated using the PCR-based QuikChange® site-directed mutagenesis method (Stratagene) as described [15] with pET-RoGACBM21 as the template, two complementary primers containing the desired mutation, and Pfu Turbo DNA polymerase (Stratagene). All constructs were transformed into competent E. coli BL21-Gold (DE3) for protein expression.
Quantitative measurement of binding to starch
The starch-binding isotherm was analysed by a saturation-binding assay as reported previously [15]. Wild-type and mutant RoGACBM21 derivatives (100 μl, 5–50 μM) in sodium acetate (50 mM, pH 5.5) were mixed with 0.1 mg of prewashed insoluble starch and incubated at 25 °C with gentle stirring for 16 h. After centrifugation at 16000 g for 10 min at 4 °C, the protein concentration of the supernatant (unbound protein) was determined by the BCA (bicinchoninic acid) protein assay, and the amount of bound protein was calculated from the difference between the initial and unbound protein concentrations. The Kd and Bmax values were determined by fitting to the non-linear regression of the binding isotherms using a standard single-site binding model.
Quantitative measurement of binding to soluble carbohydrate
Fluorescence spectrophotometry of the binding of wild-type or mutant RoGACBM21 to βCD was recorded by measuring changes in the intrinsic protein fluorescence intensity. Experiments were performed in 50 mM sodium acetate (pH 5.5) at 25 °C using a PerkinElmer LS-55 spectrophotometer. Circular and linear carbohydrates (2–20 mM) were titrated into RoGACBM21 (1 μM, 2 ml), and the fluorescence-emission spectrum was monitored at 350 nm with a fixed excitation at 280 nm. The relative changes in fluorescence intensity were plotted against the ligand concentration, and the data were fitted to a simulated curve using the appropriate equation for a single binding site.
Crystallization trials were carried out using the hanging-drop vapour-diffusion method. Both protein (1 μl) and reservoir (1 μl) solutions were mixed and equilibrated against a reservoir solution (500 μl) in Linbro plates. Initial crystallization conditions were obtained using Hampton Research Crystal Screen kits and then further optimized to obtain diffraction-quality crystals. The concentration of RoGACBM21 used for crystallization was approx. 10 mg/ml. The βCD and G7 were used at a molar ratio of 1:2 (protein/carbohydrate) to form SBD–βCD and SBD–G7 complexes respectively. Three SBD crystals grew in different conditions at 293 K. The apo-SBD crystal was crystallized in 25% PEG [poly(ethylene) glycol] 4000, the SBD–βCD crystal was grown using 18% PEG 8000 and 0.2 M zinc acetate, and the SBD–G7 crystals were grown in 30% PEG 8000 and 0.6 M ammonium sulfate.
X-ray data collection
The X-ray diffraction data were collected at beamline BL13C1 at the NSRRC (National Synchrotron Radiation Research Center, Taiwan). The data were processed and scaled using the program HKL2000 [28]. The apo-SBD, SBD–βCD and SBD–G7 crystals diffracted to 1.25, 1.8 and 2.3 Å (1 Å=0.1 nm) resolution respectively. Both apo-SBD and the SBD–βCD crystals belong to the orthorhombic P212121 space group with one molecule per asymmetric unit and the VM [29] was calculated as 1.81 Å3·Da−1 and 2.64 Å3·Da−1 respectively. The SBD–G7 crystal belongs to the monoclinic P21 space group with four molecules per asymmetric unit and the VM [29] was calculated as 2.74 Å3·Da−1 (Table 1).
Structural determination and refinement
The crystal structures of RoGACBM21 were determined using the unliganded solution structure RoGACBM21 (PDB: 2DJM) [16] as a search model by molecular replacement. The molecular replacement program MOLREP [30] was used for phase determination. Data between 8.0 and 4.0 Å and a Patterson radius of 20 Å were used to calculate the rotation and translation functions. A similar procedure was applied for the three structures and significant rotation and translation solutions were obtained. Structural model building was carried out using XTALVIEW [31], and the structural refinement was performed by CNS [32] and the CCP4 program suite [33]. The final statistics of refinement for apo-SBD and the complexes, SBD–βCD and SBD–G7, are summarized in Table 1. The co-ordinates of the RoGACBM21 structures have been deposited in the PDB as the accession numbers 2VQ4 (apo-SBD), 2V8L (SBD–βCD) and 2V8M (SBD–G7).
CBM folding topology
Several CBM superfamilies, including CBM20, CBM21, CBM25, CBM26, CBM34, CBM41 and CBM48, contain an SBD in either the N- or C-terminus [24,25,34–40]. These SBDs reveal low sequence identity but a similar immunoglobulin-like folding topology. RoGACBM21 contains 106 residues, which has low sequence identity (<15%) with AnGACBM20, as well as that of other SBD-containing CBM families. A structure-based multiple-sequence alignment of SBDs from nine representative CBM superfamilies is shown in Figure 1. The sequence alignment was carried out based on the overall structural folding of eight β strands of SBDs and the corresponding ligand-binding sites. Two types of SBD topologies, type I and type II [15], show a similar overall structure by switching the first and last β strands. Superfamilies CBM20, CBM25, CBM26 and CBM41 belong to the type I topology, whereas CBM21, CBM34 and CBM48 belong to type II topology [15].
Overall structures
The RoGACBM21 and the SBD–βCD and SBD–G7 complexes share a similar overall structure, which belongs to a typical β-sandwich fold with an immunoglobulin-like architecture [20] with approximate dimensions of 28 Å×30 Å×42 Å. The β-sandwich is symmetric and composed of eight β strands, which are antiparallel except β7 and β8. These strands are paired as four hairpin β strands, β12, β34, β45 and β67/8, and fold as a β barrel. The β barrel contains a substantial hydrophobic core that contributes the major stabilizing force to RoGACBM21. The overall structures of SBD–βCD and SBD–G7 are shown (Figures 2A and 2B). Two carbohydrate-binding sites, designated as sites I and II, were observed in both SBD–βCD and SBD–G7 complexes, in which the sugar ligands, βCD and G7, are located at almost diagonal ends of the SBD in a perpendicular orientation. Binding site I is located at Trp47 around loop β34 and site II is located at Tyr32 around loop β23. The carbohydrate-binding ratio for both SBD–βCD and SBD–G7 complexes is one sugar ligand per SBD molecule in crystal, in which the SBD shares each binding site with the symmetry-related molecule in the crystal such that two SBD molecules together hold one sugar ligand. Site I and site II are compatible to bind one sugar molecule co-operatively. In solution, the binding between the SBD and βCD is determined to be 1:2 by isothermal titration calorimetry (results not shown). The well-defined electron density map of βCD in the SBD–βCD complex is shown in Figure 2(C). The Kd (∼333 μM) of the αCD complex is much higher than that of βCD and G7 complexes [15], because the cyclic ring of the six glucose units of αCD is too small to fit the essential binding surface of the SBD, such that its binding ability is dramatically reduced.
There are four molecules per asymmetric unit in the SBD–G7 complex. The overall structures of four SBDs are very similar with RMSD (root mean square deviation) values of 0.57–1.04 Å in Cα. The key aromatic residues, Trp47, Tyr83, Tyr94 and Phe58 are in the same orientation, except for Tyr32, which is still located in the centre of G7 in the SBD–G7 complex, but tilted in a different orientation (Figure 2D). The conformations of four G7s vary; however, they tend to fold into a U shape and fit the curvature of the SBD-binding site. The major conformational difference of G7 appears to be located in the two ends of molecules.
Binding sites
Site I mainly comprises three conserved aromatic residues, Trp47, Tyr83 and Tyr94, to form a broad, flat and stable hydrophobic environment. The aromatic rings of these residues construct the curvature of the SBD-binding site and interact with the sugar rings by ideal hydrophobic stacking interactions (Figure 2C). Site I is rather rigid because Tyr83 and Tyr94 are located in β strands and Trp47 is locked by Asn46 and Asn50. Site II is generally formed by loops β23 and β45, particularly dominated by two aromatic residues, Tyr32 and Phe58. Nevertheless, both Tyr32 and Phe58 are in the proximity of the sugar ring where the van der Waals surface and the βCD are closely packed. The sugar ring of βCD is nearly parallel to the ring of Tyr32, which protrudes into the non-polar cavity of βCD and forms a hydrogen bond with βCD O27 (Figure 2C). Such unique binding is only observed in the SBD–βCD complex, where the α-glucan chains of βCD wrap around Tyr32 to stabilize the binding. Similar binding patterns were observed in CBM25, CD glycosyltransferase [41,42] and CBM48, the glycogen-binding domain of AMP-activated protein kinase [27], in which the corresponding residues in site II are Leu600 and Leu146 respectively. Another key residue in site II is Phe58, of which the hydrophobic phenyl ring forms a flat stacking interaction with the sugar ring of the glucosyl unit of βCD. Two planar rings of Tyr32 and Phe58 pack closely against the van der Waals surface of the βCD molecule and act like a clamp to pick up the βCD.
In addition to the hydrophobic interactions, numerous hydrophilic interactions between the SBD and βCD were found. In site I, residues Asn50 (ND2-O61:2.7 Å), Asn96 (ND2-O33:2.9 Å) and Asn101 (ND2-O22:2.6 Å; OD1-O32:2.9 Å) form two unique polyN loops (Figure 2E) and provide the main hydrophilic interactions to βCD. In site II, residues Asn29 (NH2-O3:2.8 Å), Lys34 (NZ-O3:2.8 Å and N-O3:3.3 Å) and Glu68 (OE1-O2:2.5 Å and OE2-O3:2.9 Å) supply several hydrogen-bond interactions with βCD. Meanwhile, these residues interact with each other by hydrogen bonds to provide a tight binding environment for βCD and contribute the forces to stabilize the SBD. The binding area of site II is small and narrow and it protrudes away from the complex more than site I.
PolyN loop
Besides the hydrophobic interaction, several asparagine residues from β34 and β78 near site I provide additional hydrophilic interactions for the carbohydrate binding. These two unique polyN loops positioned on the two sides of the sugar molecule interact with the hydroxy groups to protect the hydrophobic curvature of the binding-site groove created by Trp47, Tyr83 and Tyr94 from exposure to solvent (Figure 2E). The polyN loops consisting of consecutive asparagine residues in loop β34 (Asn46, Asn48, Asn49 and Asn50) as well as in loop β78 (Asn96, Asn97, Asn98 and Asn101) (Figure 2F) participate in sugar binding.
The significant interaction networks created in each of the polyN loops of both complexes stabilize the binding loop and facilitate sugar binding. In polyN loop β34, residues Asn46 and Asn50 directly interact with the main chains of Trp47 such that the aromatic ring of Trp47 forms hydrophobic stacking with the sugar ring (Figure 2F). Meanwhile, Asn50 and Trp47 lock together to assist the sugar binding by a hydrogen bond (Figure 2F). Furthermore, the geometry of Asn50 is squeezed out by an interaction network, Asn46–Trp47–Asn48–Asn50–Gly51–Asn52–Asn46, and the side chain of Asn50 is forced to interact with the sugar molecule (Figure 2F). Likewise, in polyN loop β78, Asn96 interacts with Ser99, Ala100 and Asn101 by three hydrogen bonds to build a βCD-binding network (Figure 2F). The orientation of Asn101 is fixed by Asn96 and Asn97 with two hydrogen bonds, as well as controlled by the adjacent residues, Ala100, Tyr102 and Gln103. As a result, Asn101 binds βCD with two hydrogen bonds (Figure 2F). A primary triangle interaction, Asn96:CO–Asn101:N–Asn97:NH2, and a periphery interaction network, Asn96–Ala100–Gln103–Tyr102–Gln95–Asn97, are produced and tight sugar binding is generated. The orientations of Asn96 and Asn101 are especially restricted and do not reveal favourable geometries in the Ramachandran plot.
Some residues in the polyN loops show very restricted geometries because these residues have to link one by one and form a series of chain interactions, which not only stabilize the binding loops, but also confirm the sugar binding. These extra interactions from the polyN loops might produce a higher sugar-binding capacity and affinity for RoGACBM21 as compared with those of other CBM superfamilies. Furthermore, the polyN loops interact with two sides of the βCD molecule to assist in turning the orientation of βCD to form hydrophobic interactions with Trp47, Tyr83 and Tyr94. Interestingly, these characteristic polyN sequences are only observed in RoCBM21 and McCBM21 (where Mc is Mucor circinelloides) (Figure 1) [13,17].
Binding affinity for starch and βCD
To elucidate the mechanism of polysaccharide binding, we examined the essential residues in the vicinity of the binding sites according to the complex structures. A number of RoGACBM21 mutants were generated and characterized by a quantitative binding isotherm and fluorescence spectroscopic analysis (Table 2) [15]. The mutant proteins contained substitutions at the aromatic residues from site I (W47A, Y83A, and Y94A) and site II (Y32A and F58A) involved in βCD and G7 binding and two additional tyrosine residues near sites I and II, Y67A and Y93A. Mutations at hydrophilic residues in site I (N50A, N96A and N101A) and site II (N29A, K34A and E68A) involved in direct hydrogen-bond interactions with ligands were also analysed.
The site I mutants (W47A, Y83A and Y94A) exhibited decreased binding affinity for starch and βCD when compared with the Kd values for wild-type RoGACBM21 (Table 2). The βCD binding cannot be detected in the W47A mutant. Furthermore, W47A, Y83A and Y94A showed a reduced Bmax for the binding capacity to starch, especially Y83A, possibly because Tyr83 is located on the inside position of the site I-binding pocket. Site I (W47A) serves as the major carbohydrate-binding site for starch. For the site II mutants (Y32A and F58A), the Kd values of Y32A for starch and βCD binding were similar to that of wild-type SBD. In addition, Y32A had less effect on the binding of soluble oligosaccharides. F58A showed an increased Kd for the binding of starch and had the highest Kd for the binding of βCD among all of the mutants; however, this mutant exhibited almost no effect on the binding capacity for starch, with a Bmax similar to that of wild-type SBD. Because Phe58 significantly affected the binding to βCD and starch, Phe58 might play a key role in site II. The Trp47- and Tyr32-binding sites were co-operative to sugar binding, because the binding affinity of the single mutant, Y32A or W47A, was partially maintained; however, the binding of the double mutant (Y32A/W47A) to insoluble starch was almost completely abolished [15]. The binding roles of Tyr32 and Trp47 can be accommodated in RoGACBM21 complexes (Figure 2C).
From the structure-based alignment (Figure 1), two additional interesting aromatic residues, Tyr67 and Tyr93 were studied. Tyr67 is conserved in most of the SBD-containing CBM superfamilies and Tyr93 is also conserved in most of the CBM21 superfamily members [13,17]. Although Tyr67 and Tyr93 do not directly interact with sugar in both the SBD–βCD and SBD–G7 complexes (Figure 2), Y67A and Y93A mutants showed apparently lower Bmax and higher Kd values (Table 2). In particular, Y67A had a considerably reduced Bmax (3.7 μmol/g), the lowest starch-binding capacity of all of the mutants. In the SBD–βCD complex, Tyr67 points into the N-terminal loop, in which the hydroxy group of Tyr67 positions around Pro4–Ser5–Ser6–Ala7 and forms a hydrogen bond with the O atom of Ala7 and stacks with Pro4 in the N terminus (Figure 3A). The mutant Y67A showed a small Bmax and it might due to the instability of the N terminus. Thus Tyr67 might play a role in the N-terminal stabilization. The overall structure of Y67A should be maintained because the binding activity of Y67A is preserved, and the secondary structure of Y67A examined by circular dichroism was intact (results not shown). The result in the present study demonstrates that Tyr67 might play an important role in governing the starch-binding capacity for RoGACBM21.
The Kd and Bmax values for mutants with substitutions at hydrophilic residues in site I (N50A, N96A and N101A) and site II (N29A, K34A and E68A) are shown in Table 2. Mutant N50A had a reduced Bmax and the highest Kd (∼10-fold higher than wild-type) for starch binding among all site I and II mutants, and this may be attributed to the loss of hydrogen-bond interactions between Asn50 and Tyr83. Asn50 might play a major role in the binding of insoluble polysaccharides and contributes to the overall integrity of site I. Mutants N96A and N101A (site I) as well as K34A and E68A (site II) exhibited inefficient βCD binding but with high Kd values due to a loss of hydrogen-bond interactions. Consequently, we suggest that these hydrophilic residues from sites I and II might play a critical role in the binding of soluble or insoluble polysaccharides, in either an individual or co-operative manner.
A continuous polysaccharide-binding path
Based on sites I and II, as well as two crucial tyrosine residues near sites I and II, Tyr67 and Tyr93, a characteristic connection was observed between two carbohydrate-binding sites (Figure 3A). From the surface electrostatic potential distribution of the SBD–βCD complex (Figure 3B), a continuous hydrophobic surface is observed formed by residues Trp47, Tyr83, Tyr94, Tyr93 and Tyr67. Furthermore, from the solvent-accessible surface of the SBD–βCD complex (Figure 3C), there are two possible accessible surface paths to link site I and site II. One path is through residues Asn50, Tyr83, Tyr94, Tyr93 and Tyr67. From the mutation data (Table 2), Y67A and Y93A mutants showed substantial effects in the starch-binding assay, Tyr67 and Tyr93 might act as a midpoint to link two binding sites and make a continuous binding path for longer chain and larger polysaccharides, or even starch. The distance of this potential polysaccharide-binding path through Trp47 (site I), Tyr93, Tyr67 and Tyr32 (site II) is in a range of 45–60 Å with an accessible surface area of approx. 1215 Å2. The other path is through hydrophilic residues, Asn50, Lys85, Glu87, Lys35 and Lys34, which are consecutive and it might be able to link sites I and II also. The continuous binding path might be important for the raw starch/insoluble polysaccharides, but not for soluble and smaller carbohydrate molecules, such as βCD and G7. During the initial binding step, sites I and II might act as two persistent binding markers for soluble/insoluble polysaccharides, after which the continuous polysaccharide-binding path may assist further binding.
Liganded and unliganded RoGACBM21
The superimposed structures of unliganded and liganded RoGACBM21 are presented in Figure 3(D). They reveal comparable overall structures, in which RMSD values between the SBD and two complexes, SBD–βCD and SBD–G7, are 0.74 and 0.98 Å respectively, and the RMSD value between the two complexes is 0.57 Å. The comparison of the residues around the sugar-binding site of unliganded and liganded SBD is shown in Figure 3(E). An apparent conformational change was observed in Tyr32, which formed a hydrogen bond with Asn29 in unliganded SBD; however, the phenyl ring of Tyr32 flipped over and reoriented to form sandwich stacking with the sugar ring upon ligand binding (Figure 3E). Surprisingly, the essential aromatic residues (Trp47, Tyr83 and Tyr94) as well as other ligand-binding residues remained in the same orientation in unliganded and liganded SBDs. Before ligand binding, numerous water molecules occupied the sugar-binding site and interacted with several hydrophilic residues, such as Lys34, Glu68, Asn50, Asn94 and Asn101. However, upon ligand binding, this water layer was replaced by a sugar molecule and the interactions were established among the sugar and hydrophilic binding residues.
These results indicated that the sugar-binding site of SBD is rather fixed and stable. As long as the β barrel scaffold is formed and the three key binding positions (Trp47, Tyr83 and Tyr94) can be preserved, the ligand binding is likely to take place. For linear polysaccharides, the Kd values of RoGACBM21 decreased with increasing ligand length and it was worse than that of βCD [15]. The Y32A mutant significantly diminished the affinity in G7 but had no effect in the short linear carbohydrates, such as maltotriose and maltotetraose [15]. Tyr32 may adjust the orientation of its phenyl ring to accommodate the conformation of a long ligand.
RoGACBM21 complexes and other CBM superfamilies
The three-dimensional structures of two CBM21 members, apo-RoGACBM21 and apo-HsCBM21 are superimposed in Figure 4(A). They share a similar overall structure except for loops β34, β45 and β56. The corresponding residues for ligand binding residues in site I (Trp47, Tyr83 and Tyr94) and site II (Tyr32 and Phe58) in the RoGACBM21 complex were also observed in HsCBM21 (where Hs is Homo sapiens) in putative site I (Trp72, Ala114 and Trp125) and site II (Phe59 and Tyr82) respectively. These residues are located in comparable locations except that the orientations of the side chains vary. We suggest that HsCBM21 might possess binding characteristics similar to that of RoGACBM21.
AnGA, a GA from Aspergillus niger [23], shares the same glycoside hydrolase characteristic as RoGA. However, the SBDs of both proteins, AnGACBM20 and RoGACBM21, fold with different topologies [15] and link with the catalytic domains through their N- and C-terminal ends respectively [23,43]. The three-dimensional structures of the two SBDs could be superimposed by switching the first and the last β strands; the structural superimposition is presented in Figure 4(B). Most CBM superfamilies contain one sugar-binding site; however, RoGACBM21 and AnGACBM20 complexes contain two binding sites [24–26,40]. RoGACBM21 binds co-operatively to sugar, whereas AnGACBM20 exhibits independent sugar binding [22]. Even though the sugar-binding location is similar in both proteins, several structural differences were noted (Figure 4B), particularly in loops β34, β45 and β78 around the sugar-binding regions, which are in completely different conformations. These distinct loops provide the unique sugar binding for RoGACBM21 and AnGACBM20.
The two binding sites of AnGACBM20 and RoGACBM21 are structurally and functionally different and the corresponding residues in AnGACBM20 are Trp543 and Trp590 for site I and Tyr527 and Tyr556 respectively. In AnGACBM20 [23], site I (Trp543) acts as the initial recognition site for starch, whereas site II (Tyr527) is capable of recognizing a range of starch strand orientations. Site I has a larger surface area, undergoes a conformational change upon sugar binding, and is proposed to act as a more specific site to lock the ligand into place. However, from the structural results and functional assay data (Table 2) for the RoGACBM21 complexes, we assume that site I (Trp47) is the essential binding site and site II (Tyr32) plays an auxiliary role as a recognition site because a conformational change and the protruded orientation of Tyr32 was clearly observed between liganded and unliganded SBDs (Figure 3E). The residues corresponding to Trp47 in most SBD superfamilies are highly conserved, except for BhCBM26 (where Bh is B. halodurans) (Figure 1). Site I reveals a larger and broader binding surface, and undergoes further conformational changes upon βCD binding.
TvAI, an α-amylase from T. vulgaris [25] that catalyses the hydrolysis of α-D-(1,4)-glucoside linkages in starch to release the α-anomer, shows different hydrolase characteristics with RoGA. However, both SBDs belong to the N-terminal starch-binding CBM superfamily and fold with the same type II topology [25,43]. Although the sequence identity and similarity between these two molecules are only 16 and 36% respectively, their structures superimposed quite well, especially the β strands. The structural superimposition of RoGACBM21 and TvAICBM34 is shown in Figure 4(C) with an RMSD of 1.6 Å in Cα. Nevertheless, structural differences were observed in loop regions, for example loops β34 and β78 (Figure 4C). Two sugar-binding sites, site-NA and site-N, found in TvAICBM34 [25] correspond to site I and site II respectively in RoGACBM21. Both sugar-binding sites were in similar orientations but two sugar molecules were in a perpendicular orientation in both proteins. Meanwhile, the functional roles of the two sugar-binding sites in both proteins were comparable: site I/site-NA binds sugar specifically and site II/site-N helps the enzyme approach starch by recognizing the starch surface.
There is a carbohydrate-binding curvature observed in several CBM families, such as TvAICBM34 [25] and BhCBM25 [24] complexes to provide the main protein–carbohydrate interactions by several key residues, for example Trp51, Tyr89 and Tyr119 in TvAICBM34 and His26, Trp34 and Trp74 in BhCBM25. This binding platform was also formed in RoGACBM21 by the corresponding residues, Trp47, Tyr83 and Tyr94. These key residues have a similar location in each protein structure, and their aromatic rings hold the sugar molecules by hydrophobic stacking interactions. Within this binding platform, the most conserved residue is Trp47 in RoGACBM21, corresponding to Trp74 in BhCBM25 and Tyr89 in TvAICBM34. This binding curvature could swing around the substrate to create a tight binding pocket and induce at least three glucose units of sugar molecules to be bent in a segmented shape for binding.
The crystal structures of unliganded and liganded RoGACBM21 (SBD–βCD and SBD–G7) were determined at 1.25, 1.8 and 2.3 Å resolutions respectively. Two carbohydrate-binding sites, I and II, were found on the surface of SBD, of which site I is a flat and broad hydrophobic-binding region created by the aromatic residues Trp47, Tyr83 and Tyr94, and site II is a protruded and narrow binding environment formed by Tyr32 and Phe58. Site I serves as the major sugar-binding site by forming an essential binding curvature for at least three glucose units. Site II may play an auxiliary role for recognizing the long-chain polysaccharides. A significant interaction network, which was created in the polyN loops unique to RoGACBM21, stabilizes the loops and confirms the sugar binding. This might produce a higher binding capacity for RoGACBM21 compared with other CBM superfamilies. An especially stable binding environment in SBD was suggested to accommodate a variety of sugars with different conformations and lengths. From the structural determination and the quantitative binding assay of RoGACBM21, a possible long-chain insoluble polysaccharide-binding path was suggested, in which the polysaccharide is bound by the specific binding sites I and II as well as a continuous binding surface with Tyr67 and Tyr93 between two sites.
This work was supported by grants from the National Science Council of Taiwan (NSC 96-2311-B-007-014 and NTHU 96N2424E1 to Y.-J. S. and NSC 96-2627-B-007-003, to M. D. T. C.). This work was partially supported by a Council of Agriculture grant, National Science and Technology Programme for Agricultural Biotechnology (97AS-1.2.1.-ST-a5, to M. D. T. C.). The RoGACBM21 clone was kindly provided by Mr Chia-Chin Sheu (Simpson Biotech, Taoyan, Taiwan). The X-ray diffraction data were carried out at the NSRRC (National Synchrotron Radiation Research Center, Taiwan).
Abbreviations: AnGA, Aspergillus niger GA; CBM, carbohydrate-binding module; βCD, β-cyclodextrin; G7, maltoheptaose; GA, glucoamylase; PEG, poly(ethylene) glycol; RMSD, root mean square deviation; RoGA, Rhizopus oryzae GA; SBD, starch-binding domain
- © The Authors Journal compilation © 2008 Biochemical Society


text: I have been a Blackberry user for a long time and I am slowly converting to Droid . One of the frustations I have run into is that I cannot figure out how to assign a buttton on the side or top to turn mute on and off during a call. I am on conference calls all the time and having to press the button on the screen doesnt work too well since the screen can lock , etc. Anyone know of a way to mute in and out with a physcial button??? I am running a rooted 3g Slide device with Cyanogen if that helps any.

subset: commoncrawl


text:
Texas Petro Index shows oil and gas industry may be leveling off
Published 12:50 pm, Monday, October 10, 2016
The Texas Petro Index is showing signs that the worst of the damage in the state's oil and gas sector may be over, or at least close to hitting bottom.
The TPI from the Texas Alliance of Energy Producers fell to 150.5 in August, a 35.2 percent drop from 232.5 the prior year. Although it is the 21st consecutive monthly decrease in the index, the drop narrowed from a 37.5 percent decline in July. The index is based on rig counts, drilling permits, oil production and other industry data.
The report comes about two years after the start of the worst collapse in oil prices since the 1980s, with crude dropping from a high of $107 a barrel in June 2014 to as low as $26 a barrel in January and February. The index is down 52 percent from its Nov. 2014 high of 313.5 and at the lowest point since 2005.
Rig counts have since increased, oil prices have stabilized and upstream oil and gas employment appears to have increased in July — the first such increase since Dec. 2014, said petroleum economist Karr Ingham, who created the index.
"These are important milestones," Ingham said in a statement. "They certainly do not suggest a return to the activity levels the state was enjoying in advance of the downturn, but they do suggest the industry is no longer hemorrhaging in terms of price, the rig count, and jobs."
Business Channel
August upstream oil and gas employment was down by more than 19 percent compared to last year and stands just above 200,000, according to his report. It's down by a third compared to the peak 306,000 who were said to be employed by the industry in Dec. 2014.
A recent announcement by members of the Organization of Petroleum Exporting Countries (OPEC) that the oil cartel will discuss production cuts has sent prices over $50 a barrel for the first time since June.


text:
Car Cut In Half In Northeast Columbus Crash
COLUMBUS, Ohio
Several people only suffered minor injuries after a northeast Columbus crash that split a car in half.
It happened at the intersection of Woodsedge and Westerville Roads around 6:20 p.m. Saturday.
The Franklin County Sheriff's Office said three cars were involved.
One of those vehicles was split in half.
At least two people were transported to area hospitals.
The Mifflin Township Police Department is continuing to investigate the cause.
Stay with 10TV and refresh 10TV.com for continuing coverage.
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Ill Tone – Bringing the Hope Back (Album) [Review]
Vancouver, BC – Since his first introduction to the habit that kick started his drug addiction, Ill Tone is clearly an individual who doesn't play it safe. So the fact that he held nothing back in his debut album, Bringing the Hope Back, shouldn't be much of a surprise.
Opening with "Only the Beginning", Ill Tone clears the air; "Screw the past I'm moving forward/ future's just around the corner/I'm no longer with Broken Logic". The next couple of tracks are filled with heavier material; he spits about his back stabbing friends and goes into great description about his drug addiction. Despite the heavy content, the beats for the first half of the album have a mellow vibe. Things take a quick turn on "Insanity"; Ill dips into the realm of electro hip-hop and although the vibe was unexpected, he kills it. Kyprios and Skryptcha join him on the next track, "If Only For A Second", to spit about the struggles they experienced as artists; it's somewhat depressing. Luckily the MC samples the cheerful beat of "The Good Lord" by Kormac to offer some encouraging advice on "Try Again". The upbeat vibe is kept on "Charlene", but unfortunately her story is one that hopefully no individual has to ever experience. After Ill reflects about life on "I'll Be", he concludes the album with a remix to Charlene by Kevin Kowal.
Bringing the Hope Back is much more than an album that you put your hands up and move side to side to; it's a therapeutic experience. The thought provoking material leaves you in a humble state of reflection. Ill Tone's decision to pour out the gloomy details of his journey allows his audience to connect with him on a personal level, which is a quality that makes a true artist.
Download the album HERE.
Review by Shaheena Azmatullah for HipHopCanada
Twitter: @ill_tone
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Earn Karma Points as part of our Rewards Program when friends become members!
The nagging little pains and soreness from running I had learned to accept began to disappear. I rarely have pain in my body now. When I do, it's normally gone after one visit to the studio.
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Your Mercedes-Benz dealership isn't the only place to buy genuine or original equipment quality repair parts to fix your CL63 AMG. The vehicle in your driveway is head and shoulders above the rest compared to all the other cars, trucks, and SUVs on the road today. Mercedes-Benz vehicle owners have become used to a certain level of a reputation for strength when driving their CL63 AMG around town. Make no mistake, you didn't buy your CL63 AMG unless you knew that you wanted best-in-class service and a trusted brand both impeccably combined into one car, truck, or SUV.
Your car or truck uses liquid coolant to maintain a constant temperature in the engine, and its thermostat keeps the coolant flowing in the right direction so that your engine can maintain a constant and ideal operating temperature. On start-up, the thermostat remains closed; bypassing the flow of coolant through your radiator. This cycles the coolant through the engine block and gets it hot. Once it reaches an ideal temperature, your Mercedes-Benz CL63 AMG thermostat then opens up and allows the coolant into the radiator. Now your coolant gets cooled as it recirculates through your Mercedes-Benz CL63 AMG engine; keeping it at a steady and controlled temperature ideal for engine performance... all thanks to the thermostat.
At Car Parts Discount, we have real customer support agents here with enough expertise to help you select the appropriate Mercedes-Benz CL63 AMG part for your needs. Don't let a broken Thermostats bring about the demise of your car, truck, or SUV, and don't risk lowering its resale value by buying anything but the highest quality repair parts. When your Mercedes-Benz needs a new replacement Thermostats, there's no need to go somewhere besides the authority, Car Parts Discount. When it comes to Mercedes-Benz CL63 AMG Thermostats parts, you can find them for 2014, 2013, 2012, 2011, 2010, 2009, 2008 models by clicking the appropriate year below.


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Question: What is the free rider problem Why does it exist What
What is the free-rider problem? Why does it exist? What are examples of free riders?
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View Solution:
Answer to relevant QuestionsWhat is meant when we say that a public good is indivisible, nonrivalrous, and nonexcludable? Keeping in mind that environmental regulation is a luxury good; would you expect American attitudes to be the same as attitudes of poor citizens of developing countries? Compare the various incentive-based environmental policies to the standards approach. What are the strengths and weaknesses of each? Who are the most likely beneficiaries of higher education? Why do low-income students encounter problems as they seek access to higher education (and often middle-income students as well)? Which do you feel is more effective in reducing poverty: government poverty programs or economic growth of the nation? How do private charities fit in? Are you conservative or liberal when it comes to addressing poverty?
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Here is CNN reporter Drew Griffin today responding to Barack Obama's Presserin which he finally made remarks on the V.A. Scandal. Griffin said he was "caught off guard" by Obama's "disconnect" from the reality of what is happening across the country by taking a, "wait a see" what more studies indicate, approach to the scandal. Griffin said there have been plenty of studies and reports since 2008 to document what is happening. But Obama is still treating the whole thing as an allegation.
Jerry Jones: Cowboys Will Stand For The Flag Or "Your Ass Will Be Off The Team"
CNN Reports Jerry Lee Lewis Died On Sunday; They Had One Job And They Messed That Up!
Oldest Christopher Columbus Statue In North America Destroyed By Vandals Citing Racism!
Joy Villa NAILS Issue About Tearing Down Statues While Strategist Looked Lost
CNN's Kate Bolduan Just Got TORCHED BY Corey Stewart About Defending ANTIFA; She Tells Him To Stop Talking
Sen. Tim Scott: Trump Should Sit Down With John Lewis To Become "Better Educated"
Small Group Of Liberty Univ Former Students Return Diplomas In Protest Of Trump
Snowflakes Hang School Flag Upside Down At Half Mass; Watch This Veteran's Response!
Model Kate Upton Is Pissed Boyfriend Didn't Win Award; Rips MLB On Twitter!
Sen. Collins: "Difficult To Say" If Trump Will Be Party's Nominee In 2020
Rapper Ice T: Manhood is Dead; you're dealing with the pussyfication of the male sex
Sep 24, 1957: Eisenhower sent 101st Airborne to Protect Nine Black Students in Arkansas | Democrats crickets!!
NAACP Tells Minorities To Steer Clear Of Missouri
After Killing Coal Jobs, Dems Want To Shut Down Government To Save Miner's Healthcare?
Jailed Russian Claims FBI Offered Him Perks If He Confessed To Hacking Hillary's Emails
Danney Williams, Bill Clinton's Alleged Love Child RIPS Megyn Kelly; There Was No DNA Test!
John McCain Angry Rand Paul BLOCKED His NDAA As He Left Town For Cancer Treatments!


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AustraliaUV - Ad-Free
by: OneSchirm • 5
All Android applications categories
by: OneSchirm • 5
No matter your complexion, UV rays have an immense impact on the health of your skin. Stay on top of current conditions using AustraliaUV, an ad-free app reporting conditions from around Australia. Select your city and see how strong the UV rays really are!
AustraliaUV provides you with an easy-to-interpret number and graphic as well as suggestions on how best to protect your skin. The app also provides an all-Australia UV forecast, skin cancer resources and a way to easily email the app developer. Current weather conditions can be accessed with just a tap.
Available cities are Adelaide, Brisbane, Canberra, Darwin, Kingston, Melbourne, Newcastle, Perth, Sydney and Townsville.
This app requires the following permissions:
Reason: To detect whether an active internet connection exists.
Reason: To access UV index data from the Australian Radiation Protection and Nuclear Safety Agency.
3rd Party Libraries:
AustraliaUV uses current data reported from around Australia to the Australian Radiation Protection and Nuclear Safety Agency. Resultant UV levels are matched against the US EPA's SunWise guidelines, developed in accordance with WHO recommendations. Though we make every attempt to accurately report UV conditions and suggest appropriate precautions, please assess the situation according to your own needs and risks.
Feature photograph: http://www.flickr.com/photos/minoru_ntt/
from 5 reviews
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This guy I have been talking to for a long time finally decided he wants to hang out with me to see where things go. He told me that he's very scared of attachment and provided an example of one time that he started getting close to someone and it made him so nervous that he ignored her until she broke up with him. We have plans to go out again soon and suddenly he has been much less talkative lately. I'm trying to feel secure in that him opening up to me about stuff was a step towards him trusting me and being able to get close, but then I'm also starting to feel panicked that he is going to do the same to me before we even see eachother again. How do I handle this without scaring him away? I don't want to bring it up and pressure him about it, because I feel like that might scare him off... But I feel like I'm going crazy worrying if he is going to get cold feet with me. What do I do?
Guys, How do I handle this without scaring him away? Should I give him space?
Have an opinion?
What Guys Said 3
It's difficult, he's the one who closes himself and if he's really paranoid about those things I don't think that there's much you can do. Maybe if you tell him that you're excited to hang out with him, act naturally, maybe you'll be able to make him do things right this time.11|00|0
If the smallest commitment scares him off then he won't able to be in a relationship with you , no offense but your wasting your time21|10|0
He's warning you who he is but you've chosen to ignore it. Your subconscious is trying to remind you and that is what is making you nervous.
You could have chosen to be with a fully avaliable guy but out of all of them, you chose the one that is the most likely to ditch you. Maybe you are the fearful one...11|00|0
It is possible you are right. I am not trying to ignore what he said... but I have liked him for a long time and he told me he has strong feelings for me and wants to be with me, but is worried he won't make me happy. I guess I am trying to give it the benefit of the doubt that I can be the exception... but I am scared of getting attached/hurt as well, despite really missing a close relationship, so maybe my choosing this guy who is so unavailable is a subconscious way to avoid attachment... Hm. Thank you.
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The Whale Wins Map
3506 Stone Way N. (N. 35th St.)
206-632-9425 | Make Restaurant Reservations | Menu
The location of The Whale Wins is marked on the map below with a green arrow. Click on it to get directions if you don't see a pop-up. Enjoy your meal at The Whale Wins!
Find out which restaurants and chefs top the list of GAYOT's 2016 Annual Restaurant Awards.


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|Sjur Nørstebø Moshagen <sjur.mosha...@kolumbus.fi>
|[RT] Document based I18n sites with Cocoon
|Fri, 09 Jul 2004 09:47:57 GMT
Yesterday Upayavira wrote: > Introducing the I18NMatcher > --------------------------- > Here's a sample sitemap snippet: > > > <map:match pattern="**.html"> > <map:match type="i18n" src="content/*/{1}.xml"> > <map:generate src="{source}"/> > <map:transform src="foo.xsl"/> > <map:transform type="i18n"> > <map:parameter name="locale" value="{locale}"/> > </map:transform> > <map:serialize type="html"/> > </map:match> > </map:match> > > Once an ordinary wildcard matcher has done its job, in comes the i18n > matcher. Its job is to see whether it can find a suitable source > document for the requested page. The * is used to symbolise the place > where the locale is to be placed. If a match is successful, it will > make sitemap variables available for the source that was found, and > the locale that matched. > > Now, this seems to be quite in keeping with the Cocoon sitemap model, > and gives some rather nice, flexible functionality. > > What do you all think? This looks great! Exactly what I need;) A couple of small questions for clarification: 1) The requested locale might be complex in the sense that it contains more information than merely the preferred language (e.g. en_GB, de_AU, sv_FI, ru_EE-KOI8). The {source} returns the best match given the available documents, but what would the {locale} be, given that {source} is not a complete match for the requested locale? One beneficial approach would IMHO be to return the "complete" locale that matched the returned document. To illustrate: The locales requested as above: en_GB, de_AU, sv_FI, ru_EE-KOI8 Available documents: content/de/foo.xml, content/sv/foo.xml {source} = content/de/foo.xml {locale} = de_AU This way it is possible for the subsequent i18n processing to cater for country-(or whatever)-specific requirements in menus etc. without the document itself necessarily being adjusted for such variation. 2) What happens if the locale of the returned document is not available in the rest of the i18n chain? There are at least two possible solutions: - just use the default locale - provide in {locale} not only the returned locale of the document, but the whole list of preferred locales _from the document match on_. In the example above that would have given: {locale} = de_AU, sv_FI, ru_EE-KOI8 Pros: This might be considered a better aproach by the user, as the further i18n processing will return locales more preferred than the default Cons: This might appear confusing to the user, as it might not seem to be any connection between the locale of the document and the locale of the surrounding widgets (menus, dates, etc.). One could even let this choice be an option, and let the webmaster/developer decide - in the end, it is up to her/him to decide whether such a situation will arise in the first place;) (by allowing document languages that are not available in other parts of the site/webapp/i18n processing). 3) How do you define the 'site default locale'? In Cocoon's existing i18n it is done through a translation file without locale tags, but that's hardly appropriate in this case. At the same time it would be nice to define the default language within Cocoon, and not rely on external settings (e.g. in the servlet, web server, etc.). Suggestions? > When I've finished implementing this, I'll go onto extend the CLI to > be able to work effectively with this kind on i18n site, enableing it > to crawl a site for each of a range of locales. But that's for another > time. Looking forward to it;))) Great work, Upayavira, keep up! Regards, Sjur
|View raw message


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Suzette's palace is where Suzette, Poodle, Mimi and Pupcake. It is located on the grasslands of Eastern Lalaloopsy Land.
The palace
It is a very luxurious palace filled with many fancy pastries and baked goods, along with very fine resting spots, stone pathways, and a beautiful fountain. Often Suzette walks by here with her favorite parasol or hand fan.
Suzette's home is very fancy and beautiful.
There are no known visitors.
Media Appearances
- Her birthday on 3D Land
- Princess Spaghetti Day
- A Little Change of Place
- In Lalaloopsy Land's map, she lives close to Jewel Sparkles and Tippy Tumblelina, however in the series, she's far away from them.


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3 South Jersey Locations
ISC & DSC are the Family place to have FUN! 3 Facilities Conveniently located in Mt. Laurel, Cherry Hill & Deptford, New Jersey. We offer Roller Skating, FunZone Giant Jungle Gyms, Inflatables, Arcade & Redemption Center. Birthday Parties, Summer Day Camps, Youth and Adult Sports and Leagues. Visit us online at www.intsports.com for more information!
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SMALL GROUPS provide a great opportunity to get to know others at the church. Small groups study the Bible, Christian books, or sermon questions, and offer fellowship and encouragement through prayer. If you are looking for a Bible Study please go to our Bible Study page.
Looking for a resource for an existing small group? Be sure to check out our small group resources in the library (enter SGR in the search window here). Other options include utilizing Growing Deeper (podcasts forthcoming) or The Bible: Plain & Simple podcasts .
Monday Morning Quarterbacks
This small group meets from 7-8am on Monday mornings in S-137 (Welcome Room).
They discuss the previous Sunday's sermon based on the questions printed at the Welcome Desk each week.
To learn more, contact Glenn Torrey or Rich Fukutaki.
Christ and Culture-Men
This group meets on Tuesday mornings from 6:30-7:45am in S-137 (Welcome Room).
They use fiction and expository literature to address spiritual issues and to find the answers God has given in the revelation of Scripture, the life of Jesus, Creation, the Holy Spirit and the history of the church. Current book- "The Son" by Philip Meyer.
To learn more, contact John Meisel.
Men's Tuesday Morning Overlake Small Group
Tuesdays, 7-8am, Jitters Coffee
This group meets at Jitters Coffee in Overlake area. They study books of the bible. Men 35-65 are welcome.
Working Men's Fellowship
This weekly group meets on Tuesdays, 7-8:55am in S-105.
They discuss how to apply the Bible to their daily lives as they study the Old & New Testaments.
Men's Tuesday Morning Small Group
Tuesdays, 7:30-9am, S-200-Conference Room
On-going Small Group. Men ages 35-65 meet Tuesday morning and have a book discussion.
Contact Jeff for more information.
Wednesday Evening Men's Small Group S. Bellevue
Wednesdays , 7-8:30pm
South Bellevue/Renton
This group meets weekly to discuss the sermon questions.
For more information contact the leader.
Men's Wednesday Morning Christian Growth Small Group
Wednesdays, 6:45-8:30 am, S-205
This is an ongoing inter-generational small group studies the Bible. Call Paul G for more information.
Women's Good Book Group
2nd Thursday, 1pm, S-105
This group meets the 2nd Thursday of the month, from September through June. This is a book discussion group welcomes all women.
Worship through Dance
Worship through Music, led by Sophie Eilers, Thursdays, UC-105, 5:30-6:30pm.
Meeting Thursdays through November 17 and then resumes first Thursday in January.
Join Sophie as she shares her heart for dance and worship.
Men's Discussion Group
Saturdays, 8-9:30, S-105
Group meets second Saturday of each month. All men are welcome. For more information contact Hank Wysong.
Home Fellowship Group in Trilogy
1st & 3rd Sundays,
Sunday evening Home Fellowship Group. This group is open to all. Presently studying the book of Romans. They alternate between studying a book of the bible
and a Christian DVD series.
For more information call Judy Martin @ 425-968-2368


text: The First Doctor Theme: Doctor Who Rarity: The Eleven Doctors Micro-Figure Set Accessories: None Year Released: 2011 The First Doctor is a Micro-Figure in the Doctor Who line. He is depicted in his outfit seen in the episode, The Three Doctors. AppearancesEdit The Eleven Doctors Micro-Figure Set Retrieved from "http://characterbuilding.wikia.com/wiki/The_First_Doctor_(The_Eleven_Doctors_Micro-Figure_Set)?oldid=6337" Ad blocker interference detected! Wikia is a free-to-use site that makes money from advertising. We have a modified experience for viewers using ad blockers Wikia is not accessible if you've made further modifications. Remove the custom ad blocker rule(s) and the page will load as expected. Categories: Doctor Who Add category Cancel Save


text:
Troubled Waters
Normally, when a major clean-up effort for a polluted site is announced-- an effort which will bring to bear the resources of both the local and federal government-- environmentalists are happy. But this is Orange County, where the eco-friendly have often seen their green hopes fade to grey (and then get paved), so the announcement yesterday at Aliso & Wood Canyons Wilderness Park that the EPA and the Army Corps of Engineers will be joining the county to detox and otherwise improve Aliso Creek was greeted with suspicion by enviromentalists.
"They promised us we would have an opportunity to bring in our own scientists for a second opinion or a peer review for this super project. And — wham, bam — we have a press conference announcing it," said Penny Elia, chairwoman of the Sierra Club's Save Hobo Aliso Task Force.
Representatives from the Surfrider Foundation, another environmental group, said they were waiting for more information.
("Federal Agencies, O.C. Plan Aliso Creek Cleanup", Los Angeles Times)
Larry McKenney, manager of Orange County's Resources and Development Management Department, assured theTimes
that environmentalists have nothing to worry about. The cleanup/flood control plans for Aliso Creek unveiled yesterday-- a sanitization plant near the bottom of the creek, and a series of 2 ft. rock steps to slow the flow of the water-- are just preliminary, McKenney said, and there will be ample opportunity for input from environmental groups during a series of public meetings which begin next month. What more could you ask for? (OK, maybe a county government with a better track record on the environment. And maybe an EPA that hasn't beenpolluted with politics
and seen its professionalism steadily evaporate over the past 5 years. And maybe some reliable assurance that the Army Corps of Engineers will take this smallish flood control project much more seriously than they took their major, decades-longresponsibilty to protect New Orleans
from flooding. Well, you're just going to have to settle for public meetings. Reassuring, no?)
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text:
Harry S. Coin
An Experienced Sales and Finance Executive
About Harry S. Coin
Throughout his career in sales and
finance, Harry S. Coin has acquired a wealth of experience in fields
such as structured finance, energy, real estate, and equity. He spent
nearly two decades as the chairman, president, and chief executive
officer of American Bank and Trust in Davenport, Iowa, where he helped
the bank increase its assets from $90 million to $750 million. Today,
Harry S. Coin works as a manager with Aristid Financial Consulting in
Illinois, and Current Energy Solutions in Bettendorf, Iowa. He also
serves as a national account executive with Unison Energy in New York
Before launching his career in finance, Harry S. Coin studied economics and political science at the University of Iowa. After completing a banking program at the Pacific Coast Banking School in Seattle, he went on to enroll at the National School of Real Estate Finance at Southern Methodist University.
In his free time, Harry S. Coin enjoys fishing, playing sports, and participating in charitable programs sponsored by his church.


text:
Hey there, time traveller!
This article was published 13/5/2013 (1558 days ago), so information in it may no longer be current.
The Selinger government is expanding the number of VLTs in Manitoba -- a move that is expected to bring in an extra $18 million a year to the government's coffers.
The Manitoba Liquor and Lotteries Corp. will soon distribute up to 500 new video lottery terminals to hotels and beverage rooms, ending a longtime moratorium on VLT expansion in such locations.
The move will bring the total number of VLTs in the province to 6,353.
The Opposition Progressive Conservatives immediately pounced on the government for preying on vulnerable Manitobans to pad its revenues when it should instead be looking for savings.
"They can't get their spending under control and they're looking for more revenue," PC Leader Brain Pallister said outside the legislature on Monday.
The Conservatives, citing the publication Canadian Gaming Digest, said Manitoba already has the highest ratio of VLTs of any province in Canada. They noted the NDP, before gaining power, once called VLTs the "crack cocaine" of gaming.
Susan Olynik, vice-president of corporate communications and social responsibility with the Manitoba Liquor and Lotteries Corp., said the decision to increase the number of VLTs in hotels and beverage rooms was "in response to consumer and site-holder demand."
She said hotels and beverage rooms will be given the opportunity to add VLTs, depending on the volume of business they currently bring in. That will occur as the machines become available. Liquor and Lotteries is raising the cap on the number of VLTs these facilities are permitted to operate to 40 from 35.
At the same time, commissions paid to site-holders will increase to 22 per cent from 20 per cent on the first $200,000 of VLT revenues. Commissions on the next $200,000 will remain at 20 per cent and then fall after that to 18.5 per cent.
The minister responsible for lotteries, Steve Ashton, said Monday most sites will benefit from the new rules -- either through more commissions or the ability to offer more VLTs. "It doesn't establish new sites," he said of the expansion. "There's no new casinos out of this."
Asked about the hypocrisy of demonizing VLTs while in Opposition and expanding their presence once in government, Ashton said the NDP is devoting more money than ever before to the treatment of gambling addictions in Manitoba. He said the Conservatives introduced VLTs to Manitoba and, one year, even cut funding to the Addictions Foundation of Manitoba.
The government has vowed two per cent of the profits from the newly merged Manitoba Liquor and Lotteries Corp. will be spent on responsible gaming and consumption.
Manitoba Liquor and Lotteries says there are now 4,482 VLTs in its commercial VLT program. That includes machines at hotels, beverage rooms, legions (404) and Tavern United (50) next to the MTS Centre. There are another 140 at Assiniboia Downs and 1,231 on Manitoba First Nations.


text:
Effaclar A.I.
Targeted Breakout Corrector
Purges imperfections in depth
Prevents residual skin marking
15 ml tube
Local breakouts in oily skin with imperfections.
Intensive local treatment that deeply purges imperfections to accelerate reabsorption and prevent residual marks:
- Niacinamide halts the evolution of outbreaks,
- The combination of piroctone olamine and glycacil purifies the bacterial source to resorb the imperfection,
- Micro-exfoliating LHA and reparative La Roche-Posay thermal spring water activate the skin's regeneration process.
Paraben-free formula.
Product efficacy
> In 2/3 of women, imperfections are resorbed within 48 hours.*
> Accelerated cellular turnover: 3.5 days gained.**
* Imperfection reabsorption kinetics: dermatologically tested on 27 women.
** Tested using dansyl chloride on 26 subjects.
Deeply purged imperfections disappear. The skin regains a perfectly homogenous surface.
Application advice
Apply locally until imperfections disappear. Effaclar A.I. can be used before applying make-up. Non-comedogenic.
La Roche-Posay recommendsSee all products in the range
3 questions to an expert
"The thin, fragile skin
around the eye contour is
the first area to show
signs of the passage of time."
Eyes are women's first concern when it comes to combating the signs of age.
Focus on...
Adult women are increasingly facing the appearance of late-onset acne.
Adult women are increasingly facing the appearance of late-onset acne.
Focus on...
Acne and its different
Acne manifests itself in blackheads and spots. It can take several forms.


text:
A Zuz (hebrew-זוז)(pl. zuzzim;hebrew-זוזים) was an ancient Hebrew silver coin struck during the Bar Kochba revolt. They were overstruck on Roman Imperial denarii or Roman provincial drachmas of Vespasian, Titus, Domitian, Trajan and Hadrian. Four Zuz, denarii or drachmas make a Shekel, a Sela or a Tetradrachm.
It has been suggested that its name is probably a corruption of the Greek Zeus which was the deity portrayed on the reverse of every drachm and tetradrachm (four drachma) of the Seleucid period and due to the prohibition of pronouncing names of idols, it was called "zuz". This name was used from the Greek era of drachmas, through the Roman era of Denarius, and then as the quarter denomination of Bar Kochba coinage. Another suggestion is that in Hebrew, the word "zuz" means "move", or "to move", so it was called "zuzzim" to show that it was constantly moving around, usually referring to the fact that Jews must give charity, or referring to the nature of money that it moves from one to another, alternating who is wealthy.
In the Talmud, the Zuz and the dinar are used interchangingly. The difference being that the Zuz originally referred to the Greek Drachma which was a quarter of the Greek Tetradrachm weighing approximately 17 grams; while the Dinar referred to the later Roman Denarii which was a quarter of the Tyrian Shekels and had the same weight as the Jewish Shekels and the Roman provincial Tetradrachms at approximately 14 grams.
The Zuz is mentioned in the Passover Haggadah in the Passover song Chad gadya, chad gadya (One little goat, one little goat); in which the lyric of dizabin abba bitrei zuzei (Which Father bought for two zuzim (half shekel) repeats at the end of every stanza.
See also
- List of historical currencies
- First Jewish Revolt coinage
- Bar Kochba Revolt coinage
- Shekel
- ma'ah
- Prutah
External links
|This article related to Jewish history is a stub. You can help by expanding it.


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What you can do if you have just been assaulted
Donate Now
Supporting survivors of sexual violence and abuse in Sussex
01273 203380
Tweets by @SurvivorsnetBtn
This group will run for 8 weeks at Survivors' Network offices in central Brighton. It will be held on Wednesday evenings from 6pm - 8pm, starting in June.
We intend the group to be small, with a maximum of 8 people, so spaces are limited.
It will be a psycho-educational and creative group, with space for reflection and discussion. We hope to create an environment where women can share and feel supported. The weekly agenda for the gorup will be decided by the participants.
For further information or to make an appointment for an initial meeting please email:
In light of the forthcoming Police and Crime Commissioner elections next month I am writing to all candidates to offer a meeting and to ask for a number of commitments relating to the elimination of Sexual and Domestic Violence and Abuse. In particular I asked them to confirm their position on:
Making ending sexual and domestic violence and abuse a priority in their Police and Crime Plan
Protecting specialist services for women and girls
Providing ring fenced funding for specialist independent Rape Crisis services
Consulting and working with the above agencies on services for
Survivors Network and LGBT Switchboard are very happy to announce that we will be launching a pilot helpline for trans survivors of sexual violence.
Our recent research into the accessibility of sexual violence recovery services for trans survivors has highlighted a lack of appropriate and accessible services for trans survivors. We are dedicated to addressing that gap and will be launching the helpline in July 2016. We are currently recruiting trans volunteers to staff this service in Brighton.
If you would like to apply, please see the advert and download the application form:
We're absolutely delighted to announce that we are recruiting two new roles, an Advocacy Support Worker and an Operations Support Worker. For more information on the roles, and how to apply click here
We're delighted to include a guest blog here from Clare Dimyon, on the subject of media reporting of rape cases
Rajeev Syal's article (4Feb) reads: Police Chief to meet Lord Britten's Widow & Apologise over "False rape claim". Later it is clear that there is no evidence of a "false rape claim" only that the Met has concluded that there is "no case to answer over a 45-year-old rape allegation four months before Brittan's death ", very different in legal terms
Sir Bernard Hogan-Howe may/may not be visiting Lord Brittan's widow later this month but he will NOT be apologising for a "false
Registered charity no.1144878 | Ltd. Company no. 07780363 | 44 1273 203380Website produced with support from American Express®
Survivors' Network is proud to be a fully accredited member of Rape Crisis England and Wales and The Survivors Trust


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Phyllo Pastry Fingers
READY IN: 20mins
Recipe by Caithi Romeo
Really quick and easy for a snack. Not gourmet but depending on any additions you apply to the recipe they could be! Very yummy though.
Ingredients Nutrition
- 4 sheets phyllo pastry
- cooking spray
- 1⁄2 cup grated cheese
- 2 tablespoons tomato paste
Directions See How It's Made
- Separate pasty sheets and spray on one side.
- Put two sheets on top of each other and spread on tomato paste.
- Sprinkle cheese onto them.
- Put other two sheets on top of each other and lay on top.
- Cut 1 inch fingers with a very sharp knife, the entire length of the pastry.
- Twist each finger so each length twists three to four times and lay on a baking sheet.
- Put in oven and cook to pastry's cooking directions.
- Yummy!


text: LOS ANGELES, April 29, 2013 (GLOBE NEWSWIRE) -- MediSwipe Inc. ( www.MediSwipe.com) (OTCBB:MWIP), a patient security solutions and partner company for the medicinal marijuana and health care industry, today announced an exclusive agreement for the sale and distribution of hemp based energy drinks and hemp based teas to medicinal dispensaries throughout the United States with Chill Drinks LLC of Miami, Florida. MediSwipe now plans to create a new distribution division to sell the popular hemp based energy drink "CHILLO" and hemp ice tea C+SWISS to all licensed medical dispensaries in 19 states and the District of Columbia. C+SWISS is popularly known as the original hemp based ice tea approved for sale in the United States, with distribution already in Whole Foods and major markets nationally. The hemp based drink comes in eco-friendly, recyclable packaging that stands out to the health conscious target customer. C+SWISS contains all natural ingredients including non-gmo beet sugar, concentrated lemon juice, hemp seed extract, black tea extract, and natural flavoring consisting of fruit and plant extracts. CHILLO, with its distinct trademark orange can, carries the chill of hemp seed extract with the added punch and perfect blend of caffeine, vitamins B6 and B12 for the ultimate drink experience. Both brands will be available in dispensaries across the country starting next week, and may be ordered for dispensaries exclusively through MediSwipe by calling 248.262.6850 or through email at firstname.lastname@example.com. "We are extremely excited to have solidified this partnership with Chill Drinks, as we believe we represent the ideal network and audience to take these products to the next level. Before our first pallets are hitting the warehouses here in Michigan, the orders from our dispensary clients are already exceeding our expectations. We are shipping to Michigan first, with products going to California and Colorado in May where we already have sales representatives writing orders. We are planning a grassroots marketing campaign, starting in Ann Arbor, at the University of Michigan with 50,000 students and a very friendly medicinal marijuana town. We plan to offer an alternative to morning coffee and the well known energy drinks with CHILLO as it provides the added benefits of hemp seed extract with a kick," stated Michael Friedman, CEO of MediSwipe, Inc.


text:
SHOW TYPE: Opera
The following is a list of songs in the show or that have been included in the show in various versions:
Main Program
Ain't It Awful, the Heat?I Got a Marble and a StarGet a Load of Thata.k.a. Gossips' TrioWhen a Woman Has a BabySomehow I Never Could BelieveGossips' QuartetIce CreamLet Things Be Like They Always WasHail to the SchoolWrapped in a RibbonLonely HouseWouldn't You Like to Be on Broadway?What Good Would the Moon Be?Moon-Faced, Starry-Eyeda.k.a. Moon Faced, Starry EyedRemember That I CareMorningCatch Me If You Cana.k.a. Children's GameThere'll Be TroubleA Boy Like YouWe'll Go Away TogetherThe Woman Who Lived Up ThereLullabya.k.a. Sleep, Baby DearI Loved Her, TooDon't Forget the Lilac Bush
Cut prior to opening
Italy in TechnicolorShe's a Gemini GirlHoroscope Songa.k.a. That's Where Our Horoscopes LieBuon Giorno, SignoreRose's Goodbye to EasterI Live for That Day
The Great Big SkyStreet Light Is My MoonlightNation of Nationsa.k.a. Melting Pot EnsembleColomboIt's the IrishA Little Swing for SwingingHot Dog WaltzThe Kids in SchoolGossips' Trio IIIMelody
© 2017 Wisdom Digital Media


text:
Yes the driver (of the school bus) was likely speeding, but we seem locked on that. Five kids were killed…. Is there no need to question administration? Like, are there seat belts for the kids? Is there ANTHING else we can do? Like add other safety measures?
Not just there… Everywhere!!! Google Streets can locate the street you are on. I'm sure we can expand that to notify the driver of the speed limit, and equip the bus to notify of excessive speed. There is ZERO need for this to happen again!
And — the Lefties??? They seem to have gotten smart. Not totally, perhaps, but I'm not hearing much about riots any more. Just think — had the tables been turned — it would have been as messy.
But I think we are growing up. Maybe.


text:
Safari Nail Trimmers (Standard)
Safari Guillotine Large Nail Trimmer The Safari Guillotine Trimmer is made from the finest quality stainless steel for a long-lasting, sharp cutting edge. Nail trimming should be a regular part of your pet's grooming. It is important to your pet's well-being to keep the nails properly trimmed.
Availability: In stock


text:
We are finally going to Egypt in November. Plan to spend a few days in Dahab and then in Cairo.
Need help to figure out which is the best way to travel from Dahab to Cairo?
I know there is bus and I know I can get a taxi ...
Thanks for all the answers


text:
Contact: Jean Weinberg
(July 27, 2006)-- Today, Congressional Candidate Jim Craig called Jeb Bradley out on the contributions he took from Exxon's PAC while Exxon reported profits of $10.36 billion in the second quarter, the second largest quarterly profit ever recorded by a publicly traded U.S. company. While Americans pay more at the pump, the five largest oil companies reported a record $110 billion in profits in 2005. Oil companies are expected to report second-quarter profits of $33.6 billion, or 32 percent more than a year earlier, according to analysts surveyed by Thomson Financial. (Associated Press 7/27/06)
"America needs a new direction that will free us from dependence on oil, end giveaways to Big Oil companies making record profits, and enact tough laws to stop price gouging," said Jim. "As energy prices rise, George Bush and Jeb Bradley continue to favor big oil and allow them to squeeze American families, farmers, businesses, and our economy."
Exxon Mobil alone raked in $36 billion in profits, the largest annual profit of any American company. The five largest oil companies have increased their annual profit by 158 percent since 2001. Energy analysts estimate that profits for these companies in the second quarter of 2006 will be $33.6 billion, 32 percent higher than the second quarter of 2005. (Based on annual profits from ExxonMobil, Shell, BP, ChevronTexaco, and ConocoPhillips company financial reports for 2001 and 2005; Associated Press 7/21/06)
Jim Craig, candidate for Congress in New Hampshire's First District, is a lifelong resident of New Hampshire and his roots in the Granite State stretch for five generations. As a veteran of the US Army, he worked for his law degree with the help from the GI Bill and joined his family's law practice. For over 20 years, he has promoted the health and well-being of families by serving on non-profit community boards. He was elected to the New Hampshire State House in 1998 and now serves as Democratic Leader. While in Concord, he worked tirelessly to create bipartisan, practical solutions for the state. As leader, he created a labor caucus that addresses working families' concerns.
Contributions Jeb Bradley received from Exxon Mobil's PAC
2005-2006 $3,000
2003-2004 $2,500
2001-2002 $5,000


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2 photos
$4,590,000 510 acres IH 37 near FM 536 Exit / 510 acres, Pleasanton, Texas 78064 - Atascosa County
Commercial / Farm & Ranch
510 acres. Property can easily be seen along IH 37 but also consist of road frontage off Tumlinston Rd., FM 1784 and Old Adams Lane. Property has 2 irrigation wells and currently has a cattle or surface lease. Price depends on size of property location. Starting at $9,000.00 per acre.
Arthur Flores ALL SEASON REALTY


text:
Show Filters
Our C W Sellors Infinity jewellery collection puts an abstract twist on ancient symbols while also evoking an ageless symbolism of eternity with the subtle infinity inspired designs. Much of the Inifinity jewellery range showcases pieces with 'links forever joined' while others are intricately crafted into a contemporary twist design that elegantly sits on the fluent curves of the body. Our Infinity jewellery and its subtle associations with eternity make each piece the ideal gift for expressing your love and commitment to your partner. All our Infinity jewellery is handmade within our workshops in Derbyshire by our talented and hardworking craftsmen.
Sign Up for Offers and Events


text:
Hey - just wanted to let all of you Sopranos devotees know -
I'll be talking about my and everyone's theories about the ending of The Sopranos in my weekly media interview by Todd Leitz on KNX1070 all news radio out of Southern California this coming Sunday morning.
Time: 7:20am Pacific time, 10:20 am Eastern time (you can do the math for the other time zones)
Streaming everywhere in the known and unknown universe live, at www.knx1070.com
Interview will be 5-10 minutes...
Useful links:
The Sopranos and the Closure-Junkies
The Sopranos, or the Tiger?
The Sopranos Ninth of Nine: The Anti-Ending Ending
The Sopranos as a Nuts-and-Bolts Triumph of Non-Network TV my 2002 article
reviews of the first eight episodes this final season: The Sopranos: First of Nine, Second of Nine, Third of Nine, Fourth of Nine, Fifth of Nine, Sixth of Nine, Seventh of Nine, Eighth of Nine
Sopranos Conference May 2008: Preliminary Announcement
reviewing 12 Monkeys, Big Little Lies, Bloodline, Bosch, Colony, Designated Survivor, Fauda, Game of Thrones, Homeland, House of Cards, Humans, Peaky Blinders, Outlander, Ray Donovan, Sense8, Somewhere Between, Stranger Things, The: Affair, Americans, Girlfriend Experience, Man in the High Castle, Missing, OA, Walking Dead; Timeless, Travelers, Twin Peaks, Vikings, Westworld +movies, music, politics
George Santayana had irrational faith in reason - I have irrational faith in TV.
Wednesday, June 13, 2007
I'll Be Talking Sopranos Ending on KNX1070 Radio this Sunday
Subscribe to: Post Comments (Atom)


text:
Let me first tell you I am just an average B2 player. Not one of those raid boss killers.
"Wheres my AR. I want to kill the guy who designed this idiot" was the first impression after playing as Krieg for a couple of hours. Its not like something went wrong. For me the decision to get this guy felt wrong. So how all this started? Lets talk.
So got the Krieg DLC soon after it was released. Watched the trailers and was really surprised at how he butchered the crap out of those rats. Started the game and reached lv5 in no time and then i hit the first wall, head first.
"Buzzaxe cooldown - 120sec". Excuse me !!! 120sec cooldown? GBX are you out of your freaking mind? 2min cooldown for action skill? what nonsense is this? Then i started checking the skills for a cooldown skills but none existed. It felt like 1st april, again. I sat there for a moment infront of the monitor with both my hands on my head wondering if this was a joke or a money grab attempt by GBX. You have a class but can't use the skill. Why? because the cooldown is freaking 2min with no cooldown skill.
Ok, enough. started playing again and soon after beating Flynt reached lv6 and guess what? 2nd wall. again head first. This time went though his skill points in a bit more details.
Hellborn - light enemies on fire and get light your ass on fire too.
Me - this is too much for 1 day. i don't want to live in this world.
Mania - increase health. add shield recharge delay.
Me - F**K me. sorry you.
Bloodlust - mag size increase for each stack.
Me - poof. finally a skill that doesn't outright kill me.
Returned to the game and kept on playing. Once in the middle resec my skill and went full on hellborn (back then i had no idea about Moxxi gun's healing) and as (un) expected death was swift and fast. Mania and reacharge delay increase = death increases even more. Back to sanctuary and straight to bloodlust. It was only after reaching reaching TVHM i figured out about Moxxi's gun and various other factor. In short, i found this wiki and at the same time joined Gearbox Forum.
(MAD) Adventures while in TVHM coming up in part 2.


text:
Iraqi-exile thriller 'Baghdad In My Shadow' begins production
EXCLUSIVE: Switzerland-Germany-UK co-pro's cast includes Zahraa Ghandour [pictured].
Baghdad-born, Swiss-based director Samir's latest feature Baghdad In My Shadow, a thriller about Iraqi exiles living in north London, starts shooting on June 8.
Written and directed by Samir, the film will shoot over eight weeks in Baghdad, Zurich, London and Cologne, Germany.
The film is produced by Joël Jent, who also produced Samir's Iraqi Odyssey.
The film is a Switzerland-Germany-UK co-production produced by Dschoint Ventschr Filmproduktion with Coin Film and Ipso Facto Productions. The latter's credits include Nicolas Winding Refn's Valhalla Rising.
The cast includes Haytham A. Ali Al Jadah (also a professor at the University of Baghdad), Iraqi newcomer Zahraa Ghandour, Kerry Fox, Meriam Abbas, Daniel Adegboyega and Felix Scott.
Have your say
You must sign in to make a comment.


text:
Nightcore/ Nightstep
Favorite Band
Favorite band....Not song.
I have WAY too many favorite songs so I just decided to do my favorite band. Nightcore or Nightstep isn't exactly a band, but it's just a techno or dubstep mix of any song that you may have heard. I prefer Nightstep because I enjoy dubstep more than techno and Nightstep is Nightcore in dub step version.


text:
Students create a book about themselves after reading the children's classic book, The Story of Ferdinand. The students' books explore diversity and what makes each person unique.
Children's Book (731)
Fiction (1,057)
Language Arts and Writing (3,673)
Literature (2,518)
Reading and Literature (3,656)
Language Arts (122)
Reading (124)
Arts and Crafts (37)
Art and Music (49)
Skill Builder (633)
Diversity (11)
Literature (95)
Activity (2,844)


text:
Query Vars and Permalinks
Query vars which are PUBLIC will be used by WP_Query to determine what page should be displayed.
Something like
/?cat=1&author=1&year=2012will show posts of category ID 1 written by author ID 1 on year 2012
The same result should be achieve from
/2012/?cat=1&author=1. But be aware of the second example, entering such URL will redirect you to
/author/something/?cat=1which I hate. Also entering
/?cat=1&author=1will break the page.
From what I've been aware of, it because such URL will giving false positive where all conditions are meet. Take the first example, it will make all of the is_date(), is_author, and is_category() return true.
THIS IS BAD! Why?
First, WordPress behave strangely with this, and doesn't seem to have consistency on it. WP will redirect people to the proper URL if they enter the query string directly. Entering /?cat=1 for instance, will redirect to /category/uncategorized. What strange is, it doesn't act the same with /?post_format=image.
Second, Archives are based on either category, tags, date, author, and now post_format. That what most people believe when design a template like me too. Thus, if you enter the URL on the first example, the loaded page is NOT a Date Archive, NOT an Author Archive, and NOT a Category Archive. It is a HOME, or Posts Page.
How do you handle this is up to you. Me? I prefer to redirect people to Search Results page whenever they manually entered multiple query_string on URL. By doing this, I'm not breaking any archive page, and not confusing the visitor.
Just my two cents
- The topic 'Query Vars and Permalinks' is closed to new replies.


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Evaluating the Written Work of Others: One Way Economics Students Can Learn to Write
The authors present a series of writing assignments that teaches students how to evaluate and critique the written economic work of others. The foundation text is McCloskey's (2000) Economical Writing. The students' dialogues with McCloskey, with each other, and with the authors of the pieces they evaluate sharpen their understanding of, and ability to use, language as an instrument of economic thought. Interviews with former students identify specific benefits from the student perspective of this approach. The authors show how the assignment series can be modified in several ways and how the general approach, as well as the foundation text, can be used in different economics courses.
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Volume (Year): 36 (2005)
Issue (Month): 1 (January)
Pages: 43-58
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