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The Level of NMDA Receptor in the Membrane Modulates Amyloid-β Association and Perforation
Alzheimer's disease is a neurodegenerative disorder that affects mostly the elderly. The main histopathological markers are the senile plaques formed by amyloid-β peptide (Aβ) aggregates that can perforate the plasma membrane of cells, increasing the intracellular calcium levels and releasing synaptic vesicles that finally lead to a delayed synaptic failure. Several membrane proteins and lipids interact with Aβ affecting its toxicity in neurons. Here, we focus on NMDA receptors (NMDARs) as proteins that could be modulating the association and neurotoxic perforation induced by Aβ on the plasma membrane. In fact, our results showed that decreasing NMDARs, using enzymatic or siRNA approaches, increased the association of Aβ to the neurons. Furthermore, overexpression of NMDARs also resulted in an enhanced association between NMDA and Aβ. Functionally, the reduction in membrane NMDARs augmented the process of membrane perforation. On the other hand, overexpressing NMDARs had a protective effect because Aβ was now unable to cause membrane perforation, suggesting a complex relationship between Aβ and NMDARs. Because previous studies have recognized that Aβ oligomers are able to increase membrane permeability and produce amyloid pores, the present study supports the conclusion that NMDARs play a critical protective role on Aβ actions in hippocampal neurons. These results could explain the lack of correlation between brain Aβ burden and clinically observed dementia.
Alzheimer's disease (AD) is the most prevalent neurodegenerative disorder in the elderly [1]. AD manifests progressively with cognitive and behavioral impairments [2] characterized by loss of memory and learning [3]. One of the major histopathological markers of AD are the senile plaques formed by amyloid-β peptide (Aβ) aggregates [4]. These Aβ aggregates produce a complex cascade of eventsthat finally lead to synaptic failure and neuronal death [5].
A common hypothesis used to explain the toxicity induced by Aβ is the formation of amyloid pores in the plasma membrane. Regarding this, several studies from our and other laboratories support the notion that membrane disruptions are induced by Aβ [6–13]. Furthermore, Aβ perforation allows the entry of small molecules and ions, such as calcium, into the cells [6, 7, 12]. This sustained calcium influx increases the release in synaptic vesicles, leading to a delayed synaptic failure produced by vesicle depletion [6–8].
It is now believed that Aβ association and toxic actions at the membrane level can be affected by the lipid and protein composition [9, 12, 14–18].N-methyl-D-aspartate receptors (NMDARs) are exc-itatory ligand gated ion channels that have been described as important for some of the toxic effects induced by Aβ, similar to that observed with other membrane proteins like cellular prion, mGluR5, nicotinic receptor, and AβPP [9, 19–25]. The glutamate NMDA receptor is a tetramer composed of different subunit combinations (NR1, NR2A-D) allowing the influx of Na+ and Ca2 + ions into the neurons resulting in excitation [26, 27]. The relationship between AD and NMDARs is widely postulated [28, 29], but the mechanisms involving this relationship are not fully understood.
One of the strongest evidence for the clinical relevance of such interactions between Aβ and NMDARs in AD is that the NMDA receptor antagonist, memantine, is used clinically in the treatment of AD [28]. In fact, other NMDAR antagonists, like (+)MK-801, or the removal of extracellular Ca2 +, reduced Aβ1–40-induced Ca2 + transients, NO production and neurotoxicity in cultured neuroblastoma cells [30]. Moreover, (+) MK-801 partially prevented the decrease in cell viability and the energy impairment induced by Aβ1–42 in HEK293 cells transiently expressing NR1/NR2A or NR1/NR2B subunits [29].
Regarding a potential interaction between Aβ and NMDAR, the data is controversial. While some authors indicate that the co-immunoprecipitation of Aβ dodecameric oligomers with NR1 and NR2A is evidence for their interaction [31], others have failed to detect binding of Aβ1–42 to any known regulatory sites on glutamate receptors [28]. Furthermore, recent data indicates that such effects of Aβ1–42 on NMDA receptors may be due to its binding to postsynaptic anchoring proteins such as PSD-95 or other membrane proteins like prion [21, 32, 33].
Here, in an attempt to clarify a potential role of NMDARs on Aβ1–42-induced neurotoxicity, we examined the ability of the peptide to associate to and disrupt plasma membranes, something that has not been studied until now. The results indicate that NMDARs are an important factor controlling Aβ neurotoxicity.
Primary cultures of rat hippocampal neurons
Hippocampal neurons were obtained from 18-day pregnant Sprague-Dawley rats and maintained for10-14 days in vitro (DIV) as previously described [34]. All animals were handled in strict accordance with NIH guidelines and approved by the Ethics Committee of the Universidad de Concepción (Concepción, Chile).
Peptide preparation and storage
Human Aβ1–42 labeled with FAM (green fluorescence, Aβ-FAM) at its N-terminus, and unlabeled peptides were purchased from Anaspec (CA, USA). The preparation and storage were performed as previously reported by our lab [6]. Briefly, Aβ1–42 was dissolved in DMSO (10 mg/ml) and stored in aliquots at –20°C. To prepare Aβ oligomers (80 μM), aliquots of the peptide (250 μg in 25 μl of DMSO) were added to 700 μl of PBS (Gibco, USA) and vertically agitated (200 RPM at 37°C) for 90 min and stored at 4°C until use. Aβ-FAM was dissolved in DMSO (4 mg/ml) and immediately stored in aliquots at –20°C.
Neuronal transfection was performed using magnetofection with the reagent Neuromag as described by the protocol provided by the manufacturer (Oz Biosciences, France). To decrease the levels of NMDARs, siRNAs for NR1 and NR2B were co-transfected with GFP (2 μg total) to visualize the transfected neurons.
To increase the levels of NMDARs, HEK cells were transfected with the plasmids NR1, NR2B and GFP (2 μg total) using lipofectamine 2000 (Invitrogen, USA) following the manufacturer's instructions. The plasmids and siRNAs used in this study were previously described [35, 36].
Electrophysiological recordings were carried out using the patch clamp technique as previously described [8, 37]. Briefly, culture media was changed for an external solution containing (in mM): 150 NaCl, 5.4 KCl, 2.0 CaCl2, 1.0 MgCl2, 10 glucose and 10 HEPES (pH 7.4). The internal solution consisted of (in mM): 120 KCl, 2.0 MgCl2, 2 ATP-Na2, 10 BAPTA, 0.5 GTP, 10 HEPES (pH 7.4). The holding potential was fixed at –60 mV and currents were acquired using a Digidata 1200 board and the pClamp10 software (Axon Instruments, Inc.). Recording pipettes were pulled from borosilicate glass (WPI, Sarasota, FL) on a horizontal puller (Sutter Instruments, Novato, CA) having a resistancebetween 5 and 10 MΩ. Perforated recordings were obtained as previously described [6, 8]. Briefly, Aβ was added to the pipette internal solution and a 5 mV pulse was used to monitor the perforation in cell attached configuration. For evoked current recordings, the experiments were performed at room temperature (20–25°C) using a membrane potential of –60 mV. Data are given as mean±S.E.M. and were obtained from more than 5 experiments. Recordings were performed in the presence of 100 nM TTX (tetrodotoxin) to inhibit action potentials.
Experiments were performed as previously described [9]. The primary antibodies used for 16 hwere: anti-MAP2, 1 : 400 (Santa Cruz Biotechnology, CA, USA); and NR2B, 1 : 200 (Covance, USA). The secondary antibodies conjugated withFITC (ExMax/EmMax = 500/517 nm), Cy3 (ExMax/EmMax = 545/570 nm), or Cy5 (ExMax/EmMax= 649/670 nm) (1 : 400, Jackson ImmunoResearch Laboratories, USA) were incubated for 2 h for fluorescent staining. Finally, samples were mounted in fluorescent mounting medium (DAKO, CA, USA) and images were obtained using a Nikon Eclipse confocal microscope (Nikon, Japan). The immunoreactivity of the proteins was quantified at primary processes with ImageJ software (NIH). Fluorescent signal was quantified as relative units (RU) using a region of interest (ROI).
Western blots
Equal amounts of proteins were separated on 10–12% SDS-PAGE gels as previously described [7, 8]. Protein bands were transferred onto nitrocellulose membranes, blocked with 5% milk, and incubated with the primary antibody anti-NR2B (1 : 1000; Covance, USA) or α-tubulin (Sigma, USA). Immunoreactive bands were detected with secondary antibodies conjugated with HRP 1 : 5000 (Santa Cruz Biotechnology, CA, USA) and visualized with ECL Plus Western Blotting Detection System (PerkinElmer, MA, USA).
Data analysis
Non-lineal analysis was performed using Prism (Graph Pad). The analysis of the cluster size, number, and fluorescence intensity were performed using the ImageJ software package (NIH, USA) and using appropriate and similar intensity thresholds for cluster resolution. Membrane charge was analyzed by integrating the transient capacitative current after subtracting the pipette capacitance. The values are expressed as mean±SEM (standard error mean). Statistical differences were determined using Student's t test or ANOVA. The experiments were performed in triplicate.
We wanted to evaluate the potential role of NMDARs in the mechanism of Aβ1–42 association and perforation of the plasma membrane. Therefore, we used experimental protocols that could either decrease or increase the levels of NMDARs in the cell membrane before performing the assays. First, we used a mild enzymatic proteolytic digestion using trypsin (0.00025%, 30 min), a serine protease widely used to remove membrane proteins [38, 39], thus decreasing the levels of NMDARs in the membrane of hippocampal neurons as detected by immunocytochemistry against the NR2B subunit of the receptor (Fig. 1A). More detailed analyses of the immunofluorescence data showed that treatment of hippocampal neurons with trypsin decreased the NMDAR puncta number in primary neuronal processes (Fig. 1B). For instance, control values were 12±1.2 punctas, and these values decreased to 6±1.10 punctas with the treatment. This result is in agreement with western blot analyses that also showed a decrease of 60±12% in NMDAR total levels after the treatment as compared to control (Fig. 1 C,D). To evaluate if the treatment was actually reducing functional membrane NMDAR in the neurons treated with trypsin for 30 min, we performed electrophysiological recordings using patch clamp techniques. The neurons were stimulated with a large concentration of NMDA (100 μM) resulting in current amplitudes of several hundred pA. The results clearly showed a significant reduction in the amplitude of the NMDA-evoked current in trypsin-treated neurons (approximately 20%) as compared to the control condition (Fig. 1E). An internal control using the NMDAR antagonist, D-AP5 (100 μM), showed that the NMDA-evoked current was completely blocked by D-AP5, indicating that the evoked current was only mediated by NMDARs (Fig. 1E). As expected, the NMDA current density (pA/pF) was also significantly lowered in trypsin-treated neurons (4.4 ± 2 pA/pF)as compared to control cells (20 ± 2 pA/pF) (Fig. 1F). No differences were found in the values of membrane capacitance indicating that the treatment did not affect the size of the neurons (Fig. 1G). These data demonstrate that the use of a very low concentration of trypsin (0.00025%) is sufficient to decrease NMDAR levels in the neuronal membrane, making this experimental protocol a good tool for further assays which need decreased levels of this ion channel receptor.
The next step was to evaluate the degree of association of oligomeric Aβ1–42 in control and trypsin treated neurons. Figure 2 shows a confocal micrograph of NR2B (red) and MAP2 (blue) together with the fluorescent signal of Aβ (1 h incubation with Aβ1–42 coupled to FAM, a green fluorescent tag)in control and trypsin-treated cells. The overall analysis of this data shows that after treating the hippocampal neurons with trypsin, the NR2B signal was reduced and that of Aβ increased (Fig. 2A). The measurement of Aβ clusters in primary neurites corroborated the increase in Aβ-FAM association together with a decrease in NMDAR on trypsin-treated neurons (Fig. 2B). A more detailed analysis indicated that the Aβ-associated clusters were much larger after the treatment (Fig. 2C). In fact, trypsin-treated neurons resulted in an increase in the size of Aβ-FAM clusters together with the number of big clusters (Fig. 2C,D).
Furthermore, to characterize the role of NMDARs in Aβ association using more specific methodologies, we used siRNA for NMDAR subunits NR1 and NR2B to selectively decrease the NMDAR levels in these hippocampal neurons. Neuronal transfections for 48 h with siRNAs also reduced the amplitude of NMDA-evoked currents (100 μM) in patch clamp recordings (Fig. 3A). For instance, the data show that treatment with the siRNA decreased the amplitude of the NMDA current from 720 ± 96 to 347 ± 63 pA (Fig. 3B), resulting in a reduction of approximately 48%. After confirming that the siRNA reduced the expression of membrane NMDARs, we incubated the neurons for 1 h with Aβ-FAM to assess the association to the neuronal membrane (Fig. 3C). The data obtained with immunocytochemistry showed that the siRNA-transfected neurons had an increase in Aβ-FAM association, incrementing the puncta number and the intensity of Aβ-FAM signal in primary processes (Fig. 3D,E), similar to the results obtained in trypsin-treated neurons.
Previous studies have shown that following Aβ association to the plasma membrane, a process of membrane damage begins that produces an increase in membrane current (perforation) and intracellular calcium levels which lead to a delayed synaptic failure [6–8, 40]. Thus, the increase in membrane association of Aβ1–42 to neurons depleted of membrane NMDARs could result in an increase in membrane damage. To determine if this was actually occurring, we performed electrophysiological experiments using perforated patch clamp recordings, as previously described [6, 8, 41] (see methods), and found that Aβ1–42 increased the peak and charge of the capacitative current after 15 minutes of application in control neurons (Fig. 4A). The effect of Aβ1–42 on the amplitude of the capacitative current, on the other hand, developed much quicker in neurons previously treated with trypsin to reduce NMDAR (Fig. 4A). The treatment, however, did not have any effect on the holding current. The time course of Aβ1–42 effects on membrane charge in control and after trypsin shows that the treatment caused an increase in the perforation onset (Fig. 4B). Data show that the approximate t1/2 of Aβ effect on the perforation was 24 min in control conditions and reduced to 17 min with the treatment. Furthermore, the analysis of the time to acquire an open configuration (perforation) in control and trypsin-treated neurons showed that the effect of Aβ1–42 was concentration-dependent, with faster effects at higher concentrations (Fig. 4C). The latter suggests that removal of membrane proteins, such as NMDARs, facilitates the membrane perforation induced by Aβ1–42.
Next, we examined if increasing NMDARs in the cell membrane could have the opposite effect, thus increasing the association of Aβ and the consequent perforation of the membrane. Thus, we overexpressed and electrophysiologically characterizedthe NMDAR subunits NR1 and NR2B in HEK cells (Fig. 5A), and glycine receptor as a control (Fig. 5B), evaluating the association of Aβ-FAM after 1 h incubation (Fig. 5C). Data showed that the Aβ-FAM association increased in the cells that overexpressed NMDARs, resulting in a reduced distance between the Aβ-FAM clusters (Control 0.55 ± 0.03, NMDA-R 0.24 ± 0.01 pixels) and an increase in their size (Control 0.46 ± 0.05, NMDA-R 0.97 ± 0.04 μm) (Fig. 5B,C), which is similar to that observed when the levels of NMDARs were decreased (Figs. 2, 3). On the other hand, overexpression of another membrane protein that is associated to inhibitory transmission, the glycine receptor (GlyR), did not produce any change in Aβ-FAM association to the plasma membrane indicating that the effect was selective for NMDARs (Fig. 5). Thereafter, we evaluated the membrane perforation induced by Aβ1–42in cells overexpressing NMDARs or GlyRs (Fig. 6A). The data obtained showed that NMDAR overexpression blocked the membrane perforation induced by Aβ1–42, while overexpression of GlyR was unable to affect the membrane charge transferred (Control 12.3 ± 5.3, Aβ 155.3 ± 14.3, NMDAR 25.4 ± 4.8, GlyRα1 135.9 ± 13.8 fC) (Fig. 6B). As a positive control for membrane perforation, we used a small peptide constructed with the native sequence of Aβ11–17 that includes the two histidine residues 13 and 14 (EVHHQKL) [42] which blocked the perforation of the membrane induced by Aβ (Fig. 6B) [6, 41, 42]. The latter suggests that NMDAR presence in the membrane of the cells increases Aβ association but interferes with the perforation induced by the peptide.
The presence of a direct or indirect interaction between Aβ and NMDARs is still under discussion, however, the role of NMDARs in AD is becoming more recognized [28]. Previous studies, for example, are focused on the use of NMDAR antagonists, like memantine or (+) MK-801 as AD treatments [29, 30]. In fact, memantine is one of the few FDA approved drugs for AD [43, 44].
In the present study, we characterized a largely unexplored area of the NMDAR relationship with Aβ regarding membrane association and subsequent perforation and that might have clinical implications. The original idea was to evaluate if the association of Aβ to the plasma membrane was affected by altering the levels of functional NMDARs, thus, cells were treated with trypsin to reduce receptor levels, or transfected with plasmids containing NMDAR subunits to increase the receptor levels. Interestingly, both treatments resulted in higher association of Aβ to the cells, suggesting a more complex role of NMDARs than expected. A simple way of explaining these results of Aβ association is what we call the "forest effect", where the membrane proteins can be likened to trees covering the ground (lipids) (Fig. 7). Thus, decreasing the level of NMDARs, suggested to be important for the interaction of Aβ with the neuronal membrane [21, 31], might be facilitating its association to lipids previously not accessible, resulting in enhanced clustering (Figs. 2 and 3). On the other hand, the increase in Aβ association after the overexpression of NMDARs might be due to enhanced direct or indirect interactions with these excitatory receptors. In agreement, it is now believed that Aβ has promiscuous membrane interactions associating to cellular prion, nicotinic receptors, AβPP, and lipids like GM1 and cholesterol, thus affecting Aβ clustering [9, 15, 17, 19–25]. Therefore, the increased Aβ association after overexpression of NMDARs could be explained by the formation of Aβ/NMDAR complexes [20, 31]. Interestingly, this effect was specific for NMDARs since overexpression of GlyRs did not affect Aβ association to the membrane. This differential effect might be relevant for the disease because GlyRs, unlike NMDARs, are inhibitory proteins that are mainly expressed in spinal cord neurons [45, 46], which are believed to be largely unaffected by the disease.
Interestingly, although decreasing or increasing NMDARs resulted in similar increases in Aβ association to the membrane (clustering), our data showed that the functional impact of altering the levels of NMDARs in the membrane was quite different. For instance, decreasing NMDARs resulted in an accelerated rate of brain membrane damage revealed as an increase in membrane current in the presence of Aβ. This is remarkable because it demonstrates differences on the impact that Aβ association has on the plasma membrane. One plausible explanation is that the fine-tuning of Aβ interaction with membrane lipids and proteins affects its capacity to form amyloid pores. Thus, a membrane devoid of some types of proteins (i.e., NMDARs) might be more sensitive to Aβ-induced damage. It was reported that increasing protein levels in the membrane produced a reduction on its fluidity [47], thus our results might also be explained by a reduced level of membrane fluidity. Nevertheless, lipid rafts can also regulate Aβ association to the membrane [15, 17, 18]. For instance, cholesterol in lipid rafts can affect Aβ association and membrane damage by decreasing membrane fluidity as a consequence of reduced phospholipid movement in the bilayer [17, 48].
On the other hand, we found that the decrease in NMDAR levels by siRNAs or trypsin also increased Aβ association. In parallel, the perforation of the membrane was faster when compared to control cells (Fig. 5). We believe that by decreasing the levels of NMDARs, Aβ associates preferentially to membrane lipids facilitating the process of perforation [15, 16, 49]. For example, it is believed that GM1 mediates Aβ association, seeding, fibrillogenesis and membrane disruption [15, 49]. These results could very well explain why some people with high levels of Aβ in the brain do not show dementia and vice versa. In other words, some proteins might be buffering the levels of toxic Aβ [50].
In conclusion, our data support the role of NMDARs as an important mediator for Aβ association and damage/perforation in the plasma memb-rane, actively participating in the membrane toxicity induced by Aβ. These results could explain the lack of correlation between brain Aβ burden and clinically observed dementia.
We thank Lauren Aguayo for revising the manuscript and for technical assistance, and Cecilia González for graphic design. This work was supported by FONDECYT grant N° 1140473. EF is a CONICYT fellow.
Authors' disclosures available online (http://j-alz.com/manuscript-disclosures/16-0170r1).
Evans DA , Funkenstein HH , Albert MS , Scherr PA , Cook NR , Chown MJ , Hebert LE , Hennekens CH , Taylor JO ((1989) ) Prevalence of Alzheimer's disease in a community population of older persons. Higher than previously reported. JAMA 262: , 2551–2556.
Faber-Langendoen K , Morris JC , Knesevich JW , LaBarge E , Miller JP , Berg L ((1988) ) Aphasia in senile dementia of the Alzheimer type. Ann Neurol 23: , 365–370.
Spires TL , Hyman BT ((2005) ) Transgenic models of Alzheimer's disease: Learning from animals. NeuroRx 2: , 423–437.
Ittner LM , Gotz J ((2011) ) Amyloid-beta and tau–a toxic pas de deux in Alzheimer's disease. Nat Rev Neurosci 12: , 65–72.
Selkoe DJ ((2002) ) Alzheimer's disease is a synaptic failure. Science 298: , 789–791.
Sepulveda FJ , Parodi J , Peoples RW , Opazo C , Aguayo LG ((2010) ) Synaptotoxicity of Alzheimer beta amyloid can be explained by its membrane perforating property. PLoS One 5: , e11820.
Parodi J , Sepulveda FJ , Roa J , Opazo C , Inestrosa NC , Aguayo LG ((2009) ) Beta-amyloid causes depletion of synaptic vesicles leading to neurotransmission failure. J Biol Chem 285: , 2506–2514.
Peters C , Fernandez-Perez EJ , Burgos CF , Espinoza MP , Castillo C , Urrutia JC , Streltsov VA , Opazo C , Aguayo LG ((2013) ) Inhibition of amyloid beta-induced synaptotoxicity by a pentapeptide derived from the glycine zipper region of the neurotoxic peptide. Neurobiol Aging 34: , 2805–2814.
Peters C , Espinoza MP , Gallegos S , Opazo C , Aguayo LG ((2015) ) Alzheimer's Abeta interacts with cellular prion protein inducing neuronal membrane damage and synaptotoxicity. Neurobiol Aging 36: , 1369–1377.
Arispe N , Rojas E , Pollard HB ((1993) ) Alzheimer disease amyloid beta protein forms calcium channels in bilayer membranes: Blockade by tromethamine and aluminum. Proc Natl Acad Sci U S A 90: , 567–571.
Arispe N , Pollard HB , Rojas E ((1996) ) Zn2+ interaction with Alzheimer amyloid beta protein calcium channels. Proc Natl Acad Sci U S A 93: , 1710–1715.
Kawahara M , Ohtsuka I , Yokoyama S , Kato-Negishi M , Sadakane Y ((2011) ) Membrane incorporation, channel formation, and disruption of calcium homeostasis by Alzheimer's beta-amyloid protein. Int J Alzheimers Dis 2011: , 304583.
Quist A , Doudevski I , Lin H , Azimova R , Ng D , Frangione B , Kagan B , Ghiso J , Lal R ((2005) ) Amyloid ion channels: A common structural link for protein-misfolding disease. Proc Natl Acad Sci U S A 102: , 10427–10432.
Lauren J , Gimbel DA , Nygaard HB , Gilbert JW , Strittmatter SM ((2009) ) Cellular prion protein mediates impairment of synaptic plasticity by amyloid-beta oligomers. Nature 457: , 1128–1132.
Kakio A , Nishimoto SI , Yanagisawa K , Kozutsumi Y , Matsuzaki K ((2001) ) Cholesterol-dependent formation of GM1 ganglioside-bound amyloid beta-protein, an endogenous seed for Alzheimer amyloid. J Biol Chem 276: , 24985–24990.
Yanagisawa K , Odaka A , Suzuki N , Ihara Y ((1995) ) GM1 ganglioside-bound amyloid beta-protein (A beta): A possible form of preamyloid in Alzheimer's disease. Nat Med 1: , 1062–1066.
Di Scala C , Troadec J-D , Lelièvre C , Garmy N , Fantini J , Chahinian H ((2014) ) Mechanism of cholesterol-assisted oligomeric channel formation by a short Alzheimerβ-amyloid peptide. J Neurochem 128: , 186–195.
Arispe N , Doh M ((2002) ) Plasma membrane cholesterol controls the cytotoxicity of Alzheimer's disease AbetaP (1-40) and (1-42) peptides. FASEB J 16: , 1526–1536.
Lorenzo A , Yuan M , Zhang Z , Paganetti PA , Sturchler-Pierrat C , Staufenbiel M , Mautino J , Vigo FS , Sommer B , Yankner BA ((2000) ) Amyloid beta interacts with the amyloid precursor protein: A potential toxic mechanism in Alzheimer's disease. Nat Neurosci 3: , 460–464.
Um Ji W , Kaufman Adam C , Kostylev M , Heiss Jacqueline K , Stagi M , Takahashi H , Kerrisk Meghan E , Vortmeyer A , Wisniewski T , Koleske Anthony J , Gunther Erik C , Nygaard Haakon B , Strittmatter Stephen M ((2013) ) Metabotropic glutamate receptor 5 is a coreceptor for Alzheimer Aβ oligomer bound to cellular prion protein. Neuron 79: , 887–902.
Um JW , Nygaard HB , Heiss JK , Kostylev MA , Stagi M , Vortmeyer A , Wisniewski T , Gunther EC , Strittmatter SM ((2012) ) Alzheimer amyloid-beta oligomer bound to postsynaptic prion protein activates Fyn to impair neurons. Nat Neurosci 15: , 1227–1235.
Fu W ((2003) ) β-amyloid peptide activates non- 7 nicotinic acetylcholine receptors in rat basal forebrain neurons. J Neurophysiol 90: , 3130–3136.
Cullen WK , Wu J , Anwyl R , Rowan MJ ((1996) ) β-Amyloid produces a delayed NMDA receptor-dependent reduction in synaptic transmission in rat hippocampus. Neuroreport 8: , 87–92.
He Y , Cui J , Lee JCM , Ding S , Chalimoniuk M , Simonyi A , Sun AY , Gu Z , Weisman GA , Gibson Wood W , Sun GY ((2011) ) Prolonged exposure of cortical neurons to oligomeric amyloid-β impairs NMDA receptor function via NADPH oxidase-mediated ROS production: Protective effect of green tea (–)-epigallocatechin-3-gallate. ASN Neuro 3: , 13–24.
Li S , Jin M , Koeglsperger T , Shepardson NE , Shankar GM , Selkoe DJ ((2011) ) Soluble Aβ oligomers inhibit long-term potentiation through a mechanism involving excessive activation of extrasynaptic NR2B-containing NMDA receptors. J Neurosci 31: , 6627–6638.
Chaffey H , Chazot PL ((2008) ) NMDA receptor subtypes: Structure, function and therapeutics. Curr Anaesth Crit Care 19: , 183–201.
Regehr WG , Tank DW ((1990) ) Postsynaptic NMDA receptor-mediated calcium accumulation in hippocampal CA1 pyramidal cell dendrites. Nature 345: , 807–810.
Danysz W , Parsons CG ((2012) ) Alzheimer's disease,β-amyloid, glutamate, NMDA receptors and memantine - searching for the connections. Br J Pharmacol 167: , 324–352.
Domingues A , Almeida S , da Cruz e Silva EF , Oliveira CR , Rego AC ((2007) ) Toxicity of beta-amyloid in HEK293 cells expressing NR1/NR2A or NR1/NR2B N-methyl-D-aspartate receptor subunits. Neurochem Int 50: , 872–880.
Le WD , Colom LV , Xie WJ , Smith RG , Alexianu M , Appel SH ((1995) ) Cell death induced by beta-amyloid 1-40 in MES 23.5 hybrid clone: The role of nitric oxide and NMDA-gated channel activation leading to apoptosis. Brain Res 686: , 49–60.
Venkitaramani DV , Chin J , Netzer WJ , Gouras GK , Lesne S , Malinow R , Lombroso PJ ((2007) ) Beta-amyloid modulation of synaptic transmission and plasticity. J Neurosci 27: , 11832–11837.
De Felice FG , Velasco PT , Lambert MP , Viola K , Fernandez SJ , Ferreira ST , Klein WL ((2007) ) Abeta oligomers induce neuronal oxidative stress through an N-methyl-D-aspartate receptor-dependent mechanism that is blocked by the Alzheimer drug memantine. J Biol Chem 282: , 11590–11601.
Lacor PN , Buniel MC , Furlow PW , Clemente AS , Velasco PT , Wood M , Viola KL , Klein WL ((2007) ) Abeta oligomer-induced aberrations in synapse composition, shape, and density provide a molecular basis for loss of connectivity in Alzheimer's disease. J Neurosci 27: , 796–807.
Aguayo LG , Pancetti FC ((1994) ) Ethanol modulation of the gamma-aminobutyric acidA- and glycine-activated Cl- current in cultured mouse neurons. J Pharmacol Exp Ther 270: , 61–69.
Bustos FJ , Varela-Nallar L , Campos M , Henriquez B , Phillips M , Opazo C , Aguayo LG , Montecino M , Constantine-Paton M , Inestrosa NC , van Zundert B ((2014) ) PSD95 suppresses dendritic arbor development in mature hippocampal neurons by occluding the clustering of NR2B-NMDA receptors. PLoS One 9: , e94037.
Sepulveda FJ , Bustos FJ , Inostroza E , Zuniga FA , Neve RL , Montecino M , van Zundert B ((2010) ) Differential roles of NMDA Receptor Subtypes NR2A and NR2B in dendritic branch development and requirement of RasGRF1. J Neurophysiol 103: , 1758–1770.
Peters C , Munoz B , Sepulveda FJ , Urrutia J , Quiroz M , Luza S , De Ferrari GV , Aguayo LG , Opazo C ((2011) ) Biphasic effects of copper on neurotransmission in rat hippocampal neurons. J Neurochem 119: , 78–88.
Lepke S , Passow H ((1976) ) Effects of incorporated trypsin on anion exchange and membrane proteins in human red blood cell ghosts. Biochim Biophys Acta 455: , 353–370.
Molloy MP , Herbert BR , Walsh BJ , Tyler MI , Traini M , Sanchez JC , Hochstrasser DF , Williams KL , Gooley AA ((1998) ) Extraction of membrane proteins by differential solubilization for separation using two-dimensional gel electrophoresis. Electrophoresis 19: , 837–844.
Peters C , Espinoza MP , Gallegos S , Opazo C , Aguayo LG ((2015) ) Alzheimer's Aβ interacts with cellular prion protein inducing neuronal membrane damage and synaptotoxicity. Neurobiol Aging 36: , 1369–1377.
Sepúlveda FJ , Fierro H , Fernandez E , Castillo C , Peoples RW , Opazo C , Aguayo LG ((2014) ) Nature of the neurotoxic membrane actions of amyloid-β on hippocampal neurons in Alzheimer's disease. Neurobiol Aging 35: , 472–481.
Diaz JC , Linnehan J , Pollard H , Arispe N ((2006) ) Histidines 13 and 14 in the Abeta sequence are targets for inhibition of Alzheimer's disease Abeta ion channel and cytotoxicity. Biol Res 39: , 447–460.
Reisberg B , Doody R , Stoffler A , Schmitt F , Ferris S , Mobius HJ ((2006) ) A 24-week open-label extension study of memantine in moderate to severe Alzheimer disease. Arch Neurol 63: , 49–54.
Doody RS , Tariot PN , Pfeiffer E , Olin JT , Graham SM ((2007) ) Meta-analysis of six-month memantine trials in Alzheimer's disease. Alzheimers Dement 3: , 7–17.
Aguayo LG , Castro P , Mariqueo T , Munoz B , Xiong W , Zhang L , Lovinger DM , Homanics GE ((2014) ) Altered sedative effects of ethanol in mice with alpha1 glycine receptor subunits that are insensitive to Gbetagamma modulation. Neuropsychopharmacology 39: , 2538–2548.
Mariqueo TA , Agurto A , Munoz B , San Martin L , Coronado C , Fernandez-Perez EJ , Murath P , Sanchez A , Homanics GE , Aguayo LG ((2014) ) Effects of ethanol on glycinergic synaptic currents in mouse spinal cord neurons. J Neurophysiol 111: , 1940–1948.
De Paillerets C , Gallay J , Alfsen A ((1984) ) Effect of cholesterol and protein content on membrane fluidity and 3 beta-hydroxysteroid dehydrogenase activity in mitochondrial inner membranes of bovine adrenal cortex. Biochim Biophys Acta 772: , 183–191.
Kawahara M , Kuroda Y ((2001) ) Intracellular calcium changes in neuronal cells induced by Alzheimer's beta-amyloid protein are blocked by estradiol and cholesterol. Cell Mol Neurobiol 21: , 1–13.
Chi EY , Frey SL , Lee KY ((2007) ) Ganglioside G(M1)-mediated amyloid-beta fibrillogenesis and membrane disruption. Biochemistry 46: , 1913–1924.
Forman MS , Mufson EJ , Leurgans S , Pratico D , Joyce S , Leight S , Lee VM , Trojanowski JQ ((2007) ) Cortical biochemistry in MCI and Alzheimer disease: Lack of correlation with clinical diagnosis. Neurology 68: , 757–763.


text:
cel·an·dine (sĕlən-dīn′, -dēn′)
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1. A perennial Eurasian herb (Chelidonium majus) having deeply divided leaves, showy yellow flowers, and yellow-orange latex. Also called swallowwort.
2. The lesser celandine.
[Middle English celidoine, from Old French, from Medieval Latin celidōnia, from Latin chelīdonia, feminine of chelīdonium, from Greek khelīdonion, from khelīdōn, swallow (from the association by ancient writers of the blossoming of the plant with the return of the swallows in spring); see ghel-1 in the Appendix of Indo-European roots.]
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subset: commoncrawl


text:
Darlington Real Estate - Find Your Perfect Home For Sale!
Darlington - Town vs. County Stats
Avg Price in Darlington: $316,300/Co. / County Avg $365,300
Avg Taxes in Darlington: $1,400/Co. / County. Avg $2,800
Avg Sq. Ft. in Darlington: 1,774/Co. / County Avg 2,609
Avg Price per/ft2 in Darlington: $178/Co. / County Avg $140
Avg Walkscore in Darlington: 15 / County Avg 22
Avg Year Built in Darlington: 1941/Co. / County Avg 1982
Avg Days on Website in Darlington: 191/Co. / County Avg 119
Darlington Real Estate Market Health
|Neighborhoods in Darlington MD


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"fresh from the factory to our stores"
Do you have an online shop?
No, the Noosa Chocolate Factory does not currently sell online or take phone orders.
What! Why not?
Our product range changes nearly every day! We make our products in very small batches depending on what cocoa, fruit & berries are in season or available. This makes it difficult to maintain an online store and keep an up to date stock list.
The Noosa Chocolate Factory typically makes 40-60kg of each product before shipping it to our stores, most products usually sell out within 2 days. Our freeze dried strawberries and Peanut butter crunch can sometimes sell out within hours of delivery!
Our philosophy is to make industry leading chocolate creations that are manufactured fresh and consumed fresh. Sometimes products are once-offs never to be seen again! We believe this is what makes us unique.
So please forgive us for not selling online, just remember to stock up when you next visit our Brisbane stores!


text:
Get a Visa® Reward Card for $300 with the latest innovations from Samsung. Buy any one or more qualifying Samsung kitchen appliances valued at a total of $3,000** or more and you'll get a Visa® Reward Card (after submitting valid Offer Claim Form and receipt).*
Valid Purchase Dates: 9/28/16 - 10/19/16
Call us at 1-866-888-5503.
We are here at all times.
24 hours a day. Every day.


text:
You need pitchers. I need pitchers. We all need pitchers.
It's a delicate balance, this fantasy baseball thing, especially down the stretch. You're close in several pitching categories. A few well-timed wins or saves that don't burst apart your ratios could be the difference between championship and Palookaville. Maybe you're bogged down with starters who need to be let go. Maybe you're ready to give up on saves and load up on innings eaters. Maybe five extra saves would mean so much to you, you're willing to drop a stud hitting reserve, and speculate on a set-up pitcher with closer aspirations.
If ever you're going to make a decisive move, now's the time. We're at the end of August, with four scant weeks to make up the ground Mike Mussina and Kip Wells have cost you. Accordingly, here are 10 pitchers owned in 25 percent or fewer of ESPN.com leagues. Depending on your squad and your standings, they might be the ticket to the fantasy nudge you so desperately need.


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Pixies – Indie Cindy
The press release for Pixies' long awaited – 23 years in fact – comeback album Indie Cindy proclaims "their songwriting chops have hardly wavered and the best may be yet to come". It doesn't take long to realise that might be a bit optimistic.
It's difficult to know how to assess this record – it stands on its own two feet well enough and makes for a perfectly enjoyable listening experience, but knowing what this band are capable of it's hard not to feel disappointed. Shorn of bassist Kim Deal, Pixies' differing composition from their heyday extends further than mere personnel changes. Gil Norton who produced Doolittle, Bossanova and Trompe le Monde is again at the helm but Black Francis' lyrics are noticeably less oblique, and the off-the-wall Buí±uel references, biblical allusions and assorted mythological and anecdotal weirdness are dialled down. The title track is a safe ballad that you couldn't imagine slotting in anywhere on Surfer Rosa or Doolittle. It's easy to accept that their sound is different, but while this could have been due to the addition of interesting new sonic elements, it is simply due to a watering down of the offbeat characteristics that made their earlier work so endearing.
There are definitely moments that rock in the conventional sense of the word – opener 'What Goes Boom' is a highlight in this department, as is 'Bagboy', perhaps the most traditionally Pixies moment here replete with Black Francis rants and Deal-esque vocal harmonies. But tracks like 'Greens and Blues', the title track, and 'Ring The Bell' are almost disconcertingly poppy and generally devoid of the energy that characterised their best work, while 'Andro Queen' hints subtly at a new direction but ultimately feels a bit pedestrian.
There's still a fair deal (no pun intended) of good material here though. 'Magdalena 318' is another high point and captures that excellent sci-fi meets surf rock combination present on much of their earlier material. Closing pair 'Snakes' and 'Jaime Bravo' – with the latter featuring some spoken word ramblings from Francis that hark back to the halcyon days of the band's first two LPs – along with the aforementioned rockier tracks, still carry enough energy to be captivating. It's no coincidence that Indie Cindy is at its best when Joey Santiago cuts loose with his guitar – a good helping of mid-song guitar freakout with a fair shake of the Black Francis rant sauce bottle never went astray in the Pixies recipe for success.


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|18 Sep 2009
Convert VC-1 to H264
Is it possible to Convert VC-1 to H264 format as some of my Blu-Ray movies with VC-1 format will not play in Media Center only H264 format will.
|My System Specs
|18 Sep 2009
Yes. Use AutoMKV to encode video in addition to tsMuxer to combine audio and video to m2ts files. PS3 can not play .dts audio unless on a physical blu-ray. So you can re-encode audio to ac3 and then add it to the original file to include both audios.
I know you asked about Media Center, but I included additional helpful info. Anything else, let me know.
In AutoMKV, the 2 pass XBOX/PS3 preset is excellent for encoding and compatible with almost all sorts of players.
AutoMKV - http://www.softpedia.com/get/Multime.../AutoMKV.shtml
tsMuxeR - http://www.videohelp.com/tools/tsMuxeR
|My System Specs
|18 Sep 2009
Maybe someone else can help further, but this is the registry code to merge to the reg to make WMC play mkv files. Perhaps it can be edited to do the same thing for your VC-1 files? I dunno.
Windows Registry Editor Version 5.00 [HKEY_CLASSES_ROOT\.ogm] "PerceivedType"="video" "Content Type"="video/x-ogm" [HKEY_LOCAL_MACHINE\SOFTWARE\Microsoft\Multimedia\WMPlayer\Extensions\.ogm] "Runtime"=dword:00000007 "Permissions"=dword:0000000f "UserApprovedOwning"="yes" [HKEY_CLASSES_ROOT\.mkv] "PerceivedType"="video" "Content Type"="video/x-matroska" [HKEY_LOCAL_MACHINE\SOFTWARE\Microsoft\Multimedia\WMPlayer\Extensions\.mkv] "Runtime"=dword:00000007 "Permissions"=dword:0000000f "UserApprovedOwning"="yes"
|My System Specs
|18 Sep 2009
Thanks torrentg I used tsMuxeR originally so I will give AutoMKV a go.
Forgot to mention I get Sound ok but no Picture
|My System Specs
|23 Aug 2011
Windows 7 Ultimate x64
MKV is only a container (an extension, not a format/codec), and VC-1 is a codec.
|My System Specs
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Founded in 1995, the Wings of Hope Breast Cancer Foundation is entirely volunteer-run, with nearly 100 per cent of donations put directly into programs that help ease the financial and psychological burdens of women and men living with breast cancer.
The Wings of Hope annual luncheon takes place Friday, Oct. 26 from 11 a.m. to 1:30 p.m. at the Hyatt Regency featuring guest speaker Wendy Mesley. Tickets are available at wings-of-hope.com, under events.
Bhawana Clark will be presenting a trunk show at the event, featuring her Wings Of Hope and other designs, and will donate 40 per cent of those sales to Wings of Hope. The jewelry can also be purchased through a pre-sale offering by contacting Diana Harris at firstname.lastname@example.com
Eight years ago, armed with a $5,000 loan, Bhawana Clark opened a business out of her Calgary home and headed to Bali, to source T-shirts, straw hats and jewelry.
In an unexpected twist for this single mom, her hottest seller became her own design for a silver pendant with mother-of-pearl.
Today, Clark's jewelry business has progressed to the point where her pieces can now be found in boutiques in Alberta, B.C., Saskatchewan and Manitoba. And, she's giving back to the community, designing three original pieces of jewelry for this year's Wings of Hope Breast Cancer Foundation luncheon.
Wings of Hope provides financial assistance to breast cancer patients for living expenses not covered by health care, such as some medications, groceries, utilities, child care, housing, transportation and other basic costs.
"In many cases when people are physically broken, they most often are financially and emotionally broken — giving (them) financial assistance while they focus on healing themselves gives them one less thing to worry about," says Clark.
"I am grateful that I have been invited by Wings of Hope to create beautiful jewelry in order for them to raise funds for this very worthy cause. I am grateful to be a part of some healing work in my life's path."
Indeed, the idea of healing is key to Clark's work. She earned a bachelor's degree in physical education from the University of Calgary, but always knew that she wanted to pursue a path that included this idea.
As Clark continued to explore and expand her jewelry designs, which led her to India, where semi-precious stones abound, she also learned of the research being done on the stones' healing energies.
"The gemstones I use in my jewelry are natural and are said to have healing properties and energies in many civilizations around the world," she says. "I personally discovered their effects as I was wearing the jewelry over the course of my first two years in business."
For the Wings of Hope pieces, Clark explains that she used Labradorite, pyrite, smokey quartz for their particular healing characteristics:
- Labradorite is a protective stone that diffuses negative energy. It's also a spiritual stone that raises consciousness, intuition and psychic gifts. It can help people get through changes in life, dispelling fear and giving strength and courage to the wearer.
- Smokey quartz is a grounding stone that helps people deal with stress in a calm manner. This stone allows people to move forward and leave behind what no longer serves them, as well as helping to clear the mind before meditation.
- Pyrite helps people to communicate more openly and honestly, allowing them to see the truth of the situation. This stone helps build self-confidence and self-esteem while blocking negative and toxic energy. This stone also increases oxygen, improving memory, circulation and overall energy and vitality.
Wings of Hope committee member Diana Harris said Clark was chosen as this year's designer because of her distinctive jewelry and her unique background.
"Bhawana is extremely generous and kind. She is also a very spiritual person and she is truly an 'angel' for Wings of Hope," Harris says. "She is very interested in helping others, so helping (people) who have breast cancer was a huge motivating factor for her in helping us raise money for WOH."
Harris says they only have a few hours to sell the jewelry, but she's confident the pieces will sell out because the contemporary, unique designs are versatile and can work with any woman's attire.
"We are thrilled to be working with Bhawana and are very proud to present this year's jewelry for the Wings of Hope event," she says.


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The coffee connoiseur is prepared to travel the extra stretch to find the tastiest drop, but happiest to discover it on their own doorstep. Hotly debated and hugely enjoyed, the perfect cup of coffee may even be the moment by which we savour our surrounds.
Our challenge was to design the packs for an international range by Robert Roberts. Seeking to capture the character and flavour of distinct countries, we chose iconic modes of transport representative of each destination, from the nifty Italian Vespa to the classic 50s Chevy of Cuba.
Striking imagery and strong colour blocking unifies the range, which is suitably stylish for display on counter tops at home.


text:
I watch with eyes full of tears as Rose makes her way up the stage, shaking visibly, even from where I'm seated. Danielle's coffin is place in the front of the stage, white and glistening in the sun that shines on it.
Quite a few people turned up for her funeral which is no surprise. On Sunday I had called the newspaper asking they place an advertisement in the paper, not wanting to be the one to ring all her friends.
As promise, her family called the relatives and they sit beside me now, sniffling as Rose begins her speech.
"Even though I hate to admit it, mum really was my best friend." People chuckle and I see her eyes brighten a little bit, helping her continue.
I don't know when she wrote her speech. I never even asked her if she wished to, assuming that she wouldn't want to because she hates public speaking. It wasn't until this morning when she handed me her notebook and asked me to check it did I know.
"You want to make a speech?" I had asked, dumbfounded. "You hate making speeches."
"Its mum's funeral," she shrugged. "It feels wrong not to."
Rose flows through her speech flawlessly, chocking up only once in a while. I can see the strength and pain behind her eyes and the struggle to keep strong. She recounts her favourite memories of her mother, like the time she was angry and they had a paint war together or the time we all tried to eat octopi, only to end up getting food poisoning and we had to stay home all day. It was Danielle's idea.
By the end of her speech, she had the room in tears. They applaud her loudly and I stand up to help her down the stairs, hugging her tightly when she reaches the bottom. Finally, she can cry.
We bury Danielle in our Church's graveyard with the senior pastor George conducting the ceremony. The sun isn't too hot at this time in the afternoon, making for a comfortable gathering of family and friends.
Rose and I cry the whole time together. We try to keep quiet but every now and then one of us will sniffle and let out a loud sob. Many relatives and friends give us reassuring pats on the back, but they do nothing to curb the pain in our hearts. There are only one's hands we desire and we'll never receive them again.
After the coffin is lowered and people say their goodbyes, Rose and I make our way back to the car with Tayla in tow. Her parents weren't able to make it so I offered to take her with us, much to Rose's gratitude.
The drive is quiet and uneventful. Rose chose to sit in the back with Tayla and they're both listening to music with earphones in. They're the old styled ones so I make a mental note to buy them both a pair of better ones for Christmas which is only a matter of weeks away now.
We drop Tayla off and make our way home. This time Rose is seated in the front beside me with her head resting on the window.
"You know," I tell her, trying to make conversation. "They're re-inventing the driverless cars."
"Telcorp?" she lifts her head.
"Yes. They think they might finally have a system that could pull it off properly this time."
"They all said that," Rose points out.
Not too long ago, the driverless car made its first appearance in the world. For the first four months everything was uneventful, but then the computer system crashed, killing up to two hundred people when the vehicles lost control. After that, the driverless car was shoved in the discarded pile of great inventions. That was the same year Telcorp first opened and began supplying new and exciting technology to the world. Of course, they immediately began designing a new and better system for the driverless car.
"I don't know, honey," I tease. "I think they might have this one perfected."
"You say that about all their inventions."
I laugh a little and she joins in.
"Maybe you can take the day off school one day and come visit the lab. One of the geniuses, Timothy, might be willing to show you around."
"Maybe," Rose shrugs. "That would be kind of cool. Can you do that though?"
"I don't see why not."
I can, but I don't tell her that. Telcorp is well known for keeping secrets. Everything in the lab and that isn't authorised for full viewing is rarely witnessed legally by anyone outside of the work force. If I speak to Riles though, maybe he could pull a few strings.
"Maybe Tayla could come to," I hint to Rose and she smiles lightly before resting her head on the window again.
We don't have to cook dinner that night. At the funeral many gave us all kinds of meals of comfort so we wouldn't have to worry about cooking for a few nights. It's almost annoying because the cooking is a gentle distraction from the sadness, but I appreciate the people's efforts to help and comfort us.
As our own memorial service to Danielle, Rose and I sit on the lounge room floor and go through some of the photo albums – pointing out old favourites and laughing a little.
"What are we going to do about mum's studio?" Rose asks at one point and I pause.
"Her friend Kirsty might buy it if she's still interested in running the place."
"Where will she get the artwork?"
"Make it herself I guess," I shrug. We both know it won't compare to Danielle's talent.
"Do you think maybe we can go in next weekend and take a painting or two? I'd love to hang some on my wall and, maybe I can save some for when I have my own place."
"Of course, sweetheart," I kiss her forehead. "We can take anything we want. Anything that was hers now belongs to us."
Rose shakes her head.
"It will always be mums."
We flick on the news later that night, interested and dreading what it might hold. As expected, there's been more terrorist attacks, mostly on the boundaries. Some are on the outer city.
"People are stupid," Rose wrinkles her nose in disgust. "They're only making things worse for themselves."
"And others, too. People forget that all their actions, every one of them, will affect those around them." I pause in thought. "I want to say sorry to you, Rose."
"What for?" she asks in curiosity.
"I'm sorry that these last few years I haven't been much of a father to you. I'm going to be honest with you, but I'm sure you already know what I'm going to say because you're smart, just like your mother."
She smiles lightly and I continue.
"I think that instead of letting my family be the meaning of my life, I let it become my job. Where I was investing my time into it, I should have been putting it towards you and your mother. Because I didn't I have missed out on so many precious memories that I could have shared with you both and so have you. I'm sorry for not being a good father to you and for you, but I promise to you I'm trying hard to fix up everything."
"Thank you," Rose cuddles against my side. "I forgive you. I'm sorry for being a brat of a teenager."
I laugh and she joins in.
"Just like your parents," I tease.
The next morning I manage to finish the brochure and eagerly set to work printing them out for Elaine to send off to all my clients. Despite yesterday's despair, I feel a sense of freedom from the heaviness. I don't want to ever let go off my wife, but I know each day I need to work hard in building a future for Rose and I. I know that's what Danielle would have wanted of me.
Tomorrow will mark one week. It already feels like forever. I don't know how I'll handle the lonely nights in bed, but I know I'll find away. After all, I have no choice.
I know I have to start planning for different things such as the clean-up of her studio, packing up her wardrobe, finding someone to clean the house and the most important, try and figure out how to be a mother to Rose.
Out of them all, the last will be the hardest. I don't want to be Danielle's replacement, nor do I want to find anyone to replace her. I have never believed in the idea of remarriage or even finding another person to love after you've lost your dearest. I believe we all have a soul-mate and I know without a doubt Danielle was mine. You can't just replace a soul-mate when there's only one.
That wouldn't be fair to Danielle or the non-existent future partner.
I bundle up the brochures and make my way outside my office to Elaine's desk. She's currently in a phone call and holds up her finger, asking me to wait. I nod and set the brochures on her desk before flicking through one of the magazines.
"Sorry about that," Elaine tells me after a moment as she sets down the phone. "The phone has been going non-stop today."
"I'm sorry to add this to your workload, but when you have time whether it's today or tomorrow, would you mind sending off these?" I push the bundle of brochures towards her. "We want people to get them on Monday."
She takes them from my hands and smiles are flirtatious grin. Today she's wearing an orange colour that goes well with her black outfit.
Quickly, I turn away to leave.
"Have I missed something?" Her voice stops me and I turn back around.
Her face is a mask of confusement and anger. Some sadness even touches her eyes.
"What do you mean?"
Her hands move to her hips and she glares at me.
"I mean one minute we're flirting each day and then suddenly you stop."
"My wife just-"
She interrupts me.
"You gave me the cold shoulder before the attack even happened that day. I can understand it now, but what did I do?"
Sighing, I pinch the bridge of my nose between my thumb and forefinger.
"You've done nothing wrong. It's my fault."
She looks taken aback but quickly regains her cold stare. I continue.
"I was and am a very selfish man. I had and still have the most amazing family in the world, but I still seeked more from you then any married man every should. I turned my back on the responsibilities of a father and spouse for outside thrills."
"Plenty of married men have mistresses," she points out and takes a step closer to me. "Plus now, you don't have to worry about being caught."
She reaches out and runs her hands over my shoulders.
I know I should pull away, but I hold my ground.
She steps forehead again and wraps her arms around my neck to rest her head on my shoulder.
"I've always been attracted to you and that's not about to go away. I know I could have pulled you away from your family, but that doesn't mean you can't have me on the side. I don't mind what I am to you, just as long as I'm something."
After a moment's pause, I slowly step back and rest my hands on her shoulders, fixing my gaze on hers through her glasses.
"I know I've lead you on ever since you've started here. You're a beautiful girl, Miss Robertson, but this has to end here. You are something to me, a friend. That's all it will ever be and all it should be."
Her face falls and she looks down.
"I'm sorry," I pat her shoulder again and after a moment she sighs with a nod.
"It's okay. I shouldn't be chasing married guys, should I?"
"I chased you back," I point out. "But you're right. You deserve so much more then to be someone's mistress or side-chick. I believe everyone has a soul-mate. No doubt yours is out there too."
She smiles at my words and I feel a budding warmth. I hope I'm making Danielle proud.
"I've dated on and off again," she shrugs. "All guys ever want is sex."
"I'm sorry I gave the impression I did."
"It wasn't an impression though, was it?" she purses her lips and then makes her way around the desk again, taking the bundle in her hand. "What was your wife like?"
"She was lovely," I tell her with a feeling of pride even though her previous words have left me almost empty. "Very creative, open-minded and smart."
"She was lucky to have you."
I shake my head.
"No, I was lucky to have her."
She smiles at my words.
"I hope I find someone like you."
"I hope you find someone better than me," I shake my head. "No good man every cheats on their partners. Don't let anyone ever do that to you."
"I'm sure your wife forgives you."
I thank her for her words and then head back into the office. I feel even lighter than before.
I have to wonder if I should change my job title to something more fitting. By the time my work time is up for the day I've only made one more sale. Part of the new job is ringing old customers to survey their opinions on the products and most of them are old with no more money to spend. I don't mind though. Surveys are some of my favourite parts of the business because you get to chat with the customers.
I store my notes away in the drawer ready to type up and hand to the marketing coordinator at a later date. They use the feedback to improve and change the products. That's why Telcorp has advanced above other companies – we truly do listen to feedback.
Just like every other day, I give Larry a pinch of fish food before making my way out the door. I smile to Elaine as I walk past and she smiles back, waving goodbye.
"See you tomorrow!"
At home I immediately crash onto the bed. There's no point trying to plan anything more for Riles and the group because we have to wait to hear from the government if we have their tick of approval or not with the idea. I can't even try planning it into further detail due to my own lack of knowledge. I know nothing about science or the human body so I suspect everything might have to be left in the hands of Thomas and any other geniuses Marcus can supply to help out. Can a needle change skin colour? Is it even possible to change people's sexuality?
The questions are what put me to sleep but thankfully my alarm wakes me up ready for Rose to arrive home. I yawn as I walk down the stairs, stretching my arms above my head, just as Rose walks through the front door.
"Hey dad."
"Hey, you. How was school today?"
"Not to bad," she leads me through the kitchen where she immediately makes a move towards the fridge. "We had a pop quiz in maths."
"Oh? How'd you go with it?"
She shrugs and looks down.
"I guess I did okay."
"It's okay if you didn't. I'm sure they understand giving the circumstances."
"I just hope they don't make me repeat the year like they've threatened to do multiple times already."
This is news to me, but I don't comment on it.
"Just do your best and whatever happens happens. There's nothing we can do about it."
She nods and eats a spoonful of yoghurt. I can see the tub is running low and I frown to myself before opening the fridge.
Besides some of the meals we had been given yesterday, there's nothing much left for us to eat.
"I guess you'll have to go shopping," Rose tells me as she peers into the fridge.
"Did you want to come with?"
She shakes her head.
"I'm really tired and I don't feel like going out."
"I'm thinking we might have to hire a housekeeper. You're too busy with school and I'm busy with work."
"If you didn't go out so late maybe you could do it?" she hints and I pat her head teasingly.
"I'll work on an ad now. I'll do some grocery shopping tomorrow."
"Okey dokey."
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Weight Loss Yoga Trailer (2011)
01 March 2011 40 mins
The stresses of daily life can lead to unhealthy habits including poor nutrition, lack of exercise and inadequate rest. Trudie Styler s Weight Loss Yoga helps you adopt a more balanced lifestyle to achieve your weight loss goals while creating a healthier body.
Watch the official Weight Loss Yoga 2011 trailer in HD below or find more Weight Loss Yoga videos on Vidimovie.


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What a difference a half millennium can make.
Since 1516, a Bavarian food purity law mandated that beer could contain only water, malts and hops. (When yeast's role in brewing was discovered in the 19th century, it was added to the list.) Known as the Reinheitsgebot, this law helped ensure quality, consistency — and a bit of stodginess.
About 500 years later, craft breweries around the world experiment with ingredients, often producing terrific new beers. With few exceptions, though, these innovations are not coming from Germany.
One exception: Sebastian Sauer of Freigeist, whose wild reinterpretations of traditional Teutonic brews have won a rabid international following.
"They're really, really good, and interesting," said Scot Blair, whose Hamilton's Tavern will host Sauer and tap seven of his beers at 3 p.m. Friday. "Interesting and German usually don't go hand-in-hand."
Freigeist, a spinoff venture from Cologne's Braustelle brewery, is where Sauer brews small experimental batches. For this label, he's made gose, a salty wheat beer; an aggressively hopped alt that alters this brew's usual malty profile; and sour porters.
Into his brew kettle, Sauer pours cherries, hibiscus, spruce tips. If he ranges well past the Reinheitsgebot, he also travels far beyond Bavaria. His current California tour began with a December appearance at North Park's Waypoint Public, then hit the Bay Area.
In Northern California, he's drawn crowds at beer bars urban (San Francisco's Mikkeller Bar, Oakland's Trappist Provisions) and suburban (Øl in Walnut Creek).
"He's a young dude, fun, and has a really good perspective on what he is doing," Blair said of Sauer, whom he met on the German's earlier visit to San Diego. "He's revamping these ancient styles."
This experimentation may continue here. Sauer had never tried his hand at a traditional British cask-conditioned beer, but Blair hopes to tap a cask of Sauer's peated porter, prepared with smoked Cara Cara orange peels.
"That's so not German," Blair said, "but he's started to get engaged with it."


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Product Description
Online Shopping Size Table: TypeXSSMLUS46810Bust88cm(34.6")91cm(35.8")93cm(36.6")98cm(38.6")Sleeve Length17cm(6.7")17cm(6.7")17cm(6.7")17cm(6.7")Shoulder37cm(14.6")37cm(14.6")38cm(15")39cm(15.4")Length80cm(31.5")81cm(31.9")82cm(32.3")83cm(32.7")TypeXL2XL3XL4XL5XLUS1214161822Bust103cm(40.6")108cm(42.5")113cm(44.5")118cm(46.5")127cm(50.0")Sleeve Length18cm(7.1")18cm(7.1")18cm(7.1")19cm(7.5")19cm(7.5")Shoulder40cm(15.7")42cm(16.5")43cm(16.9")44cm(17.3")46cm(18.1")Length84cm(33.1")85cm(33.5")86cm(33.9")87cm(34.3")87cm(34.3")Manual measurement, some errors may exist.Attention: Please note that the size table is only for reference,please choose right size according to your dressing habits such as slim or loose fit.If you like loose fit, please choose a larger size than the reference.Thanks. Package included:1*DressMore details: Disclaimer :About Size:Size may be 2cm/1 inch inaccuracy due to hand measure.These measurements are meant as a guide to help you select the correct size. Please take your own measurements and choose your size accordingly.we cannot accept responsibility for incorrect purchases. About Color:The precise color of the clothing items may vary depending on the specific monitor,the settings and the lighting conditions.The items colors depicted should only be used as an approximate guide About Review:1. we try to make sure you have a pleasant shopping experience with us, so that we could do more business with you. If there is any dissatisfaction, please email us before leaving any 1 or 2 ratings, negative feedback .We will try our best to resolve the issues. Please give us the opportunity to resolve any problem.2. If you are satisfied with our service, please leave us a 5 star positive feedback. Your recognition will make us more confident to develop business and serve you better. amazon 60% off Shopping | Glasses, cameras, dress, cloth, watches, shoes, baby dress party dress


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1. How far is the airport from the hotel?
The Renaissance Nashville is approximately a 20-minute drive from the Nashville International Airport (BNA)
2. Will all the classes take place at the Renaissance Nashville Hotel?
Yes, all sessions will take place on the premises.
3. When will the conference end?
The conference ends on Wednesday, May 24th at 2:00pm
4. When should I plan to arrive?
Hot Topic conference workshops begin on Monday, May 22 at 2:15pm, we recommend that you arrive a couple of hours prior to the workshop for registration check-in. Also please remember that in order to receive CE credits you need to attend the full session. Don't miss this chance to network in a unique and fun atmosphere.
5. How many people are you expecting?
The conference is expected to attract over 250 attendees. Conference attendees include stock plan administrators, accounting and compliance managers, financial reporting administrators and senior-level finance executives. Certent executive and support and services teams will also be there to greet you!
6. How can I justify my attendance?
Are you eager to attend Certent Summit in Nashville? Not sure your manager will approve the expense? Please click here to download a customizable letter template to help explain and justify your conference attendance to your boss.


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RARE 1922 HAWTHORNE'S WONDER BOOK - 1st Ed. ARTHUR RACKHAM ILLUSTRATED - COLOUR
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You are looking at a first edition of a lovely Arthur Rackham illustrated copy of HAWTHORNE'S WONDER BOOK. � Published in 1922 by Hodder & Stoughton. � Measures 10" by 7.5" 207 pages. Decorated endpapers. � With 24 colour Rackham illustrations (most tipped in with tissue guards) throughout the book. � The maroon red boards are in good condition and the gilt decoration on the front board is like new. � The spine however is damaged with the cloth peeling back a bit along the left edge of the spine. � also bumping to edges and corners. � Inside, the pages are clean and bright and the binding is holding though loosening. � Please feel free to email with any questions about this stunning book. really lovely illustrations by the master. � Sorry for the blurry photos - my camera is on its last legs - imagine the book in focus. Thanks for looking and good luck bidding,�


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Quick reference pocket guides in the field of medicine
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New York City, Los Angeles, Haiku, Maui, hawaii vegetarians, vegans and raw foodist events and singles dating
is a 60 year old, other / not disclosed male.
Living in New York City, Los Angeles, Haiku, Maui, hawaii
Vegan diet.
I'm a stable, intelligent, honest, loving, hard-working, fit, energetic, sincere, down to earth, sensual, affectionate, passionate, rebellious, compassionate being and do everything I can to better the planet and the lives of all creatures. I'm somewhat spiritual but definitely not religious. I love my family. I don't want to start a relationship with someone that's not vegan. If they don't get the basic right to life then they're not advanced enough for me. I also don't want to kiss someone with blood and animal fat on their lips. I'm a big animal rights/human rights/GLBT rights activist. I play music, mainly drums, most of the time. I play doumbek, djembe, bongos, percussion and guitar/bass/keys a little. I'm spending most of the next year writing and recording my own music for an album someday soon. I'll be recording in L.A., NYC and Berlin, but really prefer Maui. I also lived in L.A. a long time. I live simply with a minimal footprint on the Earth and am not a consumer for destructive corporations.
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text: Hope you guys can help. I'm having trouble finding an app that I had. I was playing about with multi con and cleared my home screen but then tried to get them all back and the only one I'm having trouble with is anycut i can't find it on the market the app symbol was a musical note (a cleff I think its called) it let me make playlists and put them in the shortcut if you can help, thanks ric


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Haptic marketing has been a buzzword for a few years now. Marketers are applying the importance of touch. "Haptic" is to touch as "Optics" are to vision. Touch is incredibly important to grabbing the attention of your audience. Increasingly, haptics have invaded the digital space. Al...October 23, 2017
Category: Resources
The Power of Touch
8 Reasons Direct Mail is Critical in B2B Marketing
By Heidi Tolliver-Walker, Industry Analyst, article from July issue of PIA's-MAG the magazine Day in and day out, you are bombarded with false messaging that direct mail is dead. The reality is that direct mail remains critical. Here are eight reasons why: 1. Direct mail doesn't require op...August 9, 2017
Postal Service's Rate Changes You Will See in 2017
November 2, 2016 Author: Renee Bingling (Re-posted from NeoPost) The U.S. Postal Service® has recently received approval from the Postal Regulatory Commission (PRC) to proceed with implementation of its January 2017 rate changes for First-Class Mail, Standard Mail, Periodical products, and Pack...December 1, 2016
Postal Reg Changes and Updated Lists
In the near future the post office will begin charging us a monthly fee if we exceed 0.8% in NCOA errors out of our total mailing quantity for each month. In order to avoid exceeding this percentage, it will become our default procedure to remove any records from the mailing which contain chargeable...November 3, 2016
Pantone's bold choice for colour of the year:
Two Colours! Rose Quartz and Serenity: Check out the colour formulas, guides, and standards....December 8, 2015
8 Great Fundraising Habits
From Fundraising Success – By Jeff Jowdy | Posted on January 14, 2015 Develop meaningful relationships with donors. Get to know your donors — each is different. Help to meet their needs and interests. Say "thank you" more than "please." Be sure that your next request ...January 19, 2015
Can a Print Publication Be Equally Effective Online? Testing the Effect of Medium Type on Marketing Communication
From Print In The Mix: By Robert G. Magee Source: Can a print publication be equally effective online? Testing the effect of medium type on marketing communications; Robert G. Magee, Virginia Tech; Marketing Letters; March 2013, Volume 24, Issue 1, pp 85-95. "Faced with budgetary pre...May 5, 2014
9 Things Great Leaders Say Every Day
BY Bill Murphy Jr. @BillMurphyJr from Inc Magazine People judge you by what you do–and by what you say. Here are nine phrases that should roll off your lips every single day. Your words are among your greatest tools. They're a window into your vision, your values and your abilities....April 14, 2014
Is NFC the next big thing in Print?
Near Field Technology (NFC) is the latest way to combine the print space with the digital space. While it's been used overseas for some time, it's just starting to gain a foot hold in America. A printed piece with an NFC tag enables a user to simply tap their phone to the piece and go [&he...November 7, 2013
Useful writing resource site
Check this out: You can copy and paste what you've written here and it will calculate your Fleish Readability Score. http://www.readability-score.com/...September 20, 2013
Personalized Marketing Drives Buyer Readiness and Sales
By Lenna Garibian From Marketing Profs Cross-channel, customer-centric marketing (AKA personalized marketing), the practice of tailoring offers and promotions to consumers across multiple touch points based on their past shopping or browsing experiences, appears to increase buyer readiness, engage...March 5, 2013


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Stainless Hero?
Is it just me, or has Elon Musk started to look like John DeLorean?
23 posts • joined 21 Jul 2010
Is it just me, or has Elon Musk started to look like John DeLorean?
Taking a bit of a risk aren't you? If GCHQ decodes this, you are TOAST...
(How embarrassing would that be?)
The Fellow is a fraud! Commercially made transistors? Pah! Get some Silicon and get doping...
The western world's capitalist system is steadily increasing the gap between the 'haves' and the 'have nots'. My comment was a simple reductio ad absurdem argument. You though, seem to be one of the believers who will chant "There is no alternative!" as the world slides down the crapper..
The real problem seems to be that Global Capitalism will end with a few Trillionaires and everyone else crawling around in the mud convincing themselves that "there is no alternative!"
In typical UK MOD bungling style, it was going to have steam catapults, then it wasn't, then some fancy electro-magnetic linear motor catapult, then nothing. Multiply this sort of faffing about for every system aboard and I wonder it hasn't cost ten times its already staggering cost. Still six years to finish it, so plenty of opportunities to add / delete / add again countless bits of technology. . . .
Happy memories of covering the patio doors with 'Post-it' notes as we kept a record of how to go from room to room - Creak!
I bought 30 - odd Amstrads for my school at a time when CUB monitors and BBC masters were the norm. They proved amazingly tough and reliable whereas the BBCs were a nightmare. We used PCW's as upgrades from typewriters and CPC 6128s to introduce computing. At some point we built (kit?) mice interfaces and used DMP printers. 'Protext' was a fabulous word processor and simple 'coding' done via LOGO. Happy memories.
Quite; there's nothing like high speed Porn...
Today I learnt a gun can be 'safe' and 'useful'.
Working Sonic Screwdriver - Pozidrive screwdriver with a whistle built in to the handle.
"Embrace the change. With more people listening, it can only get better."
No it won't.
DAB is an old system abandoned by the rest of the World. We need the Government to acknowledge their mistake and adopt DAB+ with more thought of firmware updates in the future.
It's Silicon. (You may however have a radio polishing fetish - who am I to judge.)
Modern cyber-war is one thing; why illustrate it with a picture of a Bristol Bloodhound missile from the 1950s?
Those pron films are the best!
or connected even...
$17.8 million seems very reasonable; have they taken inflation into account?
A distinct improvement on my DVD copy with pin-sharp graphics (though the 'dodgy' artwork in Bulldog scene still grates.) Audio seems over cooked though with alarming surges and stereo image leaping about at times. A colourful, one-off featuring the Fab Four.
The U.S. government is run by and for big corporations; they will do anything in their considerable power to keep the population hooked on their products.
Amoral and jaw - droppingly ignorant - the tomato is a fruit!
Or even space odyssey . . .
FM / Internet radio - fine; but DAB+?
Is this PURE's attempt to push the UK government into adopting a half-way sensible digital system?
Other Companies have been developing Autogyros (Gyroplanes?) for some time -
Celier have Rotax powered Gyros that are being assessed by Police forces.


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Among the methods used — the most primitive and dirty provocation
In the US city of Greenville (Mississippi) «unknown» set fire to the Methodist church for black writing on the wall «Vote for Trump.» Federal Bureau of Investigation (FBI) that refused to cover the newly discovered crime candidate for US president from the Democratic Party, Hillary Clinton, launched an investigation into an act of vandalism. This was announced by Eric Simmons — mayor of Greenville, 78 percent of whom are negros.
FBI task to harassment which joined US President Barack Obama, is now spending all of your time to save the Clinton election campaign is extremely simple. To understand the direction in which to conduct the search for «vandals», should ask only one question: who benefits? The answer is obvious. Those who want to customize the blacks, who are already historically more supportive of Democrats than Republicans, against Trump. But the goal, as it often happens, justifies the means. But it can be said, the motto of the ex-Secretary of State, this is how she and her entourage and act in life.
The investigation makes it easier is that Mississippi Governor Phil Bryant — a Republican and, of course, a supporter of Trump. He has assured the public that the instigators of the church and the authors of the provocative inscription, «will soon be found and respond with justice» for their actions.
The official representative of the FBI in Mississippi, Brett Carr said that his department «is working closely with the authorities at the city, state and country,» although he admitted that this case will not be easy.
To help the investigation, a reward of 11 thousand dollars for help in tracking down the author of the inscription «Vote for Trump.» And, as the chief of the city police department Delano Wilson, although no specific suspects at the moment, certain strings in the investigation already.
Why Trump sling mud?
While in Mississippi are investigating, campaign headquarters Donald Trump have to expose this crime prior provocation, arranged with the same goals — further customize blacks against the Republican candidate for the White House. He has to deny and disown publication supported by the Ku Klux Clan Crusader newspaper, which seems to be good at it to work.
«Trump and his campaign condemn any manifestation of hatred This publication is disgusting, do not express the views of the views of tens of millions of Americans who have joined together to support our campaign.», — Said the billionaire assistant US weekly Time.
And this is far from the first time: the American public are trying to instill the idea that Trump and his supporters — low, unworthy people, America is a shame that it is necessary to get round: sophistication, intelligence, humanity and Trump — this is inconsistent. Be careful, if you suspect someone that you support it.
Well, to give the impression that it is supposedly so employed person Democrats regularly staged violent brawl at campaign rallies Trump: beat political activists, police, bleeding. To have an impression: but what else can you expect from the loons, Trump supporters that revealed this way their relations? But soon, if not stop Trump, these people will already be a threat for the whole country.
However, not all so simple. In his famous Gettysburg Address billionaire he said that, as the American authorities have established, the money for these antics paid thugs Democratic National Committee and the Clinton campaign headquarters. And he is confident that «due to the fraudulent system (in the country — approx Constantinople.), Is likely to them nothing for it will not be.»
These above-mentioned structures are, in his words, and the appearance of many women who, through the media regularly accuse Trump «sexual harassment» and «sexism». He has no doubt that they are well paid for it: «We'll find out later in court is waiting for this.!»
Lies materialized in action
It is characteristic that these stories, mostly, of course, made-up, lead to the fact that lying is squared, and its object is already sending real black mark. Recently, the attacker, who identified himself as the first hit it, shattered personal pick Trump name of the star on the Walk of Fame in Hollywood, Los Angeles (California). As reported by the US media, «political» vandal explained his trick desire to break the brass plate with the name and surname of the billionaire in the center of the star, to sell it at auction and give the proceeds to women, which allegedly offended Trump.
But for those who is not impressive, invent stories that Trump does not pay taxes (if it tried, it would have been a political corpse, this little family of the Clintons in this respect everything is getting away with) that is associated with the Russian government and even a «coordinated» with him on the campaign trail, his mysterious people exchange messages with the Russian Alfa-Bank, and the like.
Undoubtedly, we are talking not only about a dirty campaign to discredit the billionaire, but also attempts to divert attention from the many uncomfortable questions that would not hurt to ask itself Clinton. For example, she does not know how to keep state secrets and is a boon for spyware, it has in its immediate environment is extremely suspicious persons involved is presumed terrorists in the Middle East, through the Clinton Foundation receives money from foreign donors …
In this sense, particularly injurious to Trump is the very last days: all that he blamed on the eve of the vote, to deny he had no time. This is true, of course, and for Clinton. The difference between them is only in the fact that it, as we see, is accused in the case.
Double standards
In what position they are unequal, however, are located.
«Reports that the FBI is investigating the activities of the Clinton Foundation, shows the true scale of the corrupt Clinton machine,» — he said in this regard, adviser to Republican Jason Miller. He expressed regret at the same time that the Ministry of Justice «are not permitted to open an investigation until the completion of the elections.» Miller stressed that the excitement for the week against the former Secretary of State of two investigations — on the new «e-mail scandal» and the Foundation — enough for her «full suspension» from further participation in the election campaign.
Instead, Obama attacks the FBI director James Komi, because he let the late, but still interested in Clinton's activities. Moreover, the outgoing president traveled to campaign for her to Florida, one of the United States in key states in the election — in hopes of securing his former employee voice black voters.
Why is it necessary Trump?


text:
Looking to learn more about our trucks? You've come to the right place.
The details on this truck are listed out below for you to browse, but let's talk about you.
Ever thought of the cab as the driver's office? Maybe you should. Your drivers spend hours behind the wheel and it's your job to keep them safe and alert. You do that by keeping them comfortable. This means thoughtful ergonomics, 325° visibility, and an HVAC system that just won't quit. It's a cab with room to maneuver and space to keep organized. It's a cab that won't leave them exhausted at the end of their day. Spec'ing your truck with the drivers in mind means more productivity every single day.
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Here are the details on City of Dallas's Autocar truck.
Body Company
Line of Business
Automated Side Loader
ISL9 '13 270HP Diesel
3000 6 speed
Front Axle
Meritor MFS-14 Steer Axle
Front Tire
Goodyear 12R22.5H
Rear Axle
Dana S30-90
Rear Tire
315 Goodyear
Drive Configuration


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The Sphinx from Mystery Men said it best (original quote by Benjamin Franklin).
A few months back, I facilitated a lesson planning session for a group of online tutors. During this session, we've discussed how important it is for a facilitator/trainer/educator to have a step-by-step guide on how to systematically approach a learning session. We've discussed three points as to why having a well-crafted lesson plan is beneficial – no, a necessity, to the educator.
The Lost Art of Lesson Planning


text: 1. Is there a full size keyboard and/or mouse that can connect to the htc evo? 2. Is there an app that can read text from pictures like microsoft onenote? 3. Is there an app that can convert text talk? So I can listen to m notes and/or scanned notes.


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Product Groups
Getting Started
Intraday Data
Subscription Based Data
This paper reviews 10 of the most frequently used CTA indexes, serving as a key resource for the industry. Prior to the writing of this paper, information on CTA indexes was not easy to find.
CME Group is the world's leading and most diverse derivatives marketplace. The company is comprised of four Designated Contract Markets (DCMs). Further information on each exchange's rules and product listings can be found by clicking on the links to CME, CBOT, NYMEX and COMEX.
CME Group / Chicago HQ
Phone: +1 312 930 1000
Toll Free (US Only): +1 866 716 7274
© 2017 CME Group Inc. All rights reserved.


text: United States President Barack Obama (R) walks with President Hu Jintao of China during a State arrival ceremony on the South Lawn of the White House, Wednesday, January 19, 2011 in Washington, DC. Obama and Hu are scheduled to meet in the Oval Office later in the day, hold a joint press conference and attend a State dinner. .Credit: Mark Wilson / Pool via CNP


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» Entertainment
Online casinos have really revolutionized the way people gamble. Now anyone from any part of the world can participate and walk away with thousands, even millions in cash prizes without having to invest much. One of the most popular online gambling website right now is Jackpotjoy. Jackpotjoy is famous for its wide varieties of games that are both attractive and very rewarding. This article shows you how you can enjoy your time the most playing games from this website.
Set aside a budget
Before you start playing, the first thing you need to do is set aside a budget. If you play different games from Jackpotjoy, it is important you determine the amount you are willing to spend in each one of them. It is important to note that while it is possible to walk away with cash prices from this website, losses are also a reality and it is a good idea to always come prepared. Setting aside the amount you want to use in this online casino will ensure you dont spend more than you can afford and that your experience here is fun.
Dont play with your winnings
One of the most common mistakes people make when playing games from Jackpotjoy is get greedy. They want to win more and will take their winnings and use them to play more games. While it is possible to increase your earnings playing with the extra money you get, it is also possible to lose it all and walk home with nothing. You need to be wise so that you live to gamble another day.
Never chase your losses
This is another common mistake with those who play games from Jackpotjoy. Most people, after losing while gambling will try and chase their losses. This, more often than not leads to more disasters where gamblers lose even more money, with some getting bankrupt within a few hours. This should never happen to you. Accept defeat so that you avoid more damage to your finances.
Have a strategy
Before you sit down on your computer, it is important to know which games you want to play. Go for the Jackpotjoy game you are familiar with and if you are feeling like trying something new, make sure you dont invest too much until you are fully skilled. Two of the most popular online gambling games in Jackpotjoy are Bingo and Slot machines. These games are not only easy to play but have very high chances of winning. In order to enjoy these two or any other games on Jackpotjoy, you need to formulate a plan. For instance, to increase your chances of winning with slots, you will first need to find out the hot and cold slots.
Have fun
Most people forget the fact that as much as online casinos are meant for gambling, to truly enjoy the experience, you have to make it as fun as possible. When playing from Jackpotjoy, remove the emotional part and just have fun. This will improve your chances of winning as you will have a more open mind to try out new things.
Darren Knuth is a big fan of Red or Black and Jackpotjoy games.