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Episodic ataxia type 1 (EA1) is a Shaker-like channelopathy characterized by continuous myokymia and attacks of imbalance with jerking movements of the head, arms, and legs. Although altered expression and gating properties of Kv1.1 channels underlie EA1, several disease-causing mechanisms remain poorly understood. It is likely that Kv1.1, Kv1.4, and Kvβ1.1 subunits form heteromeric channels at hippocampal mossy fiber boutons from which Zn2+ ions are released into the synaptic cleft in a Ca2+-dependent fashion. The sensitivity of this macromolecular channel complex to Zn2+ is unknown. Here, we show that this heteromeric channel possesses a high-affinity (<10 μM) and a low-affinity (<0.5 mM) site for Zn2+, which are likely to regulate channel availability at distinct presynaptic membranes. Furthermore, the EA1 mutation F184C, located within the S1 segment of the Kv1.1 subunit, markedly decreased the equilibrium dissociation constants for Zn2+ binding to the high- and low-affinity sites. The functional characterization of the Zn2+ effects on heteromeric channels harboring the F184C mutation also showed that this ion significantly 1) slowed the activation rate of the channel, 2) increased the time to reach peak current amplitude, 3) decreased the rate and amount of current undergoing N-type inactivation, and 4) slowed the repriming of the channel compared with wild-type channels. These results demonstrate that the EA1 mutation F184C will not only sensitize the homomeric Kv1.1 channel to extracellular Zn2+, but it will also endow heteromeric channels with a higher sensitivity to this metal ion. During the vesicular release of Zn2+, its effects will be in addition to the intrinsic gating defects caused by the mutation, which is likely to exacerbate the symptoms by impairing the integration and transmission of signals within specific brain areas.
- shaker channel gating
- episodic ataxia type 1
- Xenopus laevis cocytes
episodic ataxia type 1 (EA1) is an autosomal dominant neurological disorder affecting central and peripheral nerve functions. The disease is characterized by symptomatic attacks of imbalance and uncontrolled movements that may be triggered by physical or emotional stress (3, 8, 12, 22, 60). The duration of the attacks of ataxia varies: they may occur several times a day or once a year. Although the symptoms vary between and within families, two symptoms are always observed: an ataxic gait during attacks and myokymia. The latter symptom is characterized by continuous muscle activity that can be detected as a rhythmic electromyography activity with a pattern of repeated duplets and multiplets (12, 22). Genetic linkage studies have localized the EA1 locus to chromosome 12p13 (36). Subsequently, the KCNA1 gene, which encodes for the delayed-rectifier K+ channel Kv1.1, has been identified as underlying EA1 (11). All affected individuals possess heterozygous point mutations at positions highly conserved among the voltage-dependent K+ channel superfamily (10, 11). Dominant-negative effects or haploinsufficiency may underlie altered K+ channel function in EA1-affected individuals (65). Most EA1 channels are functional and, depending on the location within the Kv1.1 subunit, produce homomeric channels with biophysical properties that differ from those of wild-type channels (7, 9, 15, 16, 19, 31, 55, 66, 70). Although several lines of evidence indicate that defective delayed rectifier channels underlie EA1, many molecular mechanisms have not been thoroughly characterized (33). Recently, a mouse model of EA1 has been generated by introduction of the V408A mutation into the mouse Kv1.1 gene. These animals showed impaired motor performance and altered cerebellar GABAergic transmission from the basket cells to the Purkinje cells (27).
When EA1 subunits coassemble with wild-type subunits or with subunits belonging to the same K+ channel subfamily (e.g., Kv1.2), the resulting heteromeric proteins are endowed with partial EA1 phenotypes (16, 46). Individuals carrying the F184C mutation showed generalized motor seizures, in addition to typical EA1 symptoms (11, 60). This phenylalanine-to-cysteine substitution is located at the COOH-terminal portion of the first transmembrane domain (S1) of the Kv1.1 subunit. In previous studies, we and others showed that the F184C mutation shifted the half-maximal activation voltage (V1/2) of Kv1.1 homomeric channels ∼25 mV to more depolarized potentials, and slowed the time constants of activation by nearly twofold (1, 9, 14, 65). Recently, we also showed that this mutation sensitizes homomeric Kv1.1 channels to extracellular Zn2+ (14). However, we did not investigate whether this mutation also affects the Zn2+ sensitivity of channels composed of Kv1.4, Kv1.1, and Kvβ1.1 subunits, inasmuch as these subunits are likely to form a macromolecular channel complex in several brain structures in which Zn2+ plays a role (24, 44, 48, 50, 51, 59, 62).
Specific neurons are able to store Zn2+ within vesicles. Their release into the synaptic cleft is regulated by the frequency and intensity of the stimuli by means of Ca2+-dependent mechanisms. Neuron excitability is affected by Zn2+, which modulates the activity of selected members of ligand- and voltage-gated ion channels (for reviews see Refs. 20, 25, 40, 54, 56, 57). The location of this ion and its release pattern have been established across the brain by use of Zn2+ fluorophores, radiolabeled Zn2+, and selective chelators (20, 21, 30, 35, 58, 63). In particular, Zn2+ is released from mossy fiber terminals in the hippocampus (4, 61, 64). A growing body of evidence indicates that Kv1.1, Kv1.4, and, Kvβ1.1 subunits form heteromeric channels at hippocampal mossy fiber boutons (44, 48, 49, 51, 53, 59). Moreover, patch-clamp recordings from such presynaptic terminals suggest that this macromolecular channel complex regulates the activity-dependent spike broadening of hippocampal mossy fiber boutons and, as a consequence, the amount of neurotransmitter released during high-frequency stimuli (23). Thus an altered Zn2+-mediated modulation of heteromeric channel activity may exert a remarkable impact on the hippocampal integration and transmission of signals.
Here we show that the EA1 mutation F184C will not only sensitize the homomeric Kv1.1 channel to extracellular Zn2+, but it will also endow heteromeric channels, composed of human Kv1.1, Kv1.4, and Kvβ1.1 subunits, with a higher sensitivity to Zn2+, which also significantly modifies several biophysical properties of the mutated channel.
Molecular biology.
The human β-subunits were kindly provided by Prof. Olaf Pongs (Institut für Neurale Signalverarbeitung, Center for Molecular Neurobiology Hamburg, Universität Hamburg, Hamburg, Germany), and the human Kv1.4 subunit was cloned in our laboratory. To study the activity of channels composed of two human Kv1.1 and two human Kv1.4 subunits, the relevant cDNAs were linked as dimers (15). To concatenate the human Kv1.4 wild-type subunits with the episodic ataxia human Kv1.1 subunits as dimers, the stop codon of the first subunit was removed and a linker encoding 10 glutamine residues was inserted between the last codon of the 5′-subunit coding sequence and the initiator codon of the following subunit. This was achieved using a sequential PCR protocol similar to that used by Horton et al. (29). Briefly, junctions were generated by overlap extension of the PCR primers that also encoded the glutamine linker. The dimers were constructed in pGEMA, a modified version of pGEM9zf (Promega), with a stretch of 40 A-T base pairs inserted just upstream of the Tth111I site. The QuikChange Site-Directed Mutagenesis kit (Stratagene) was used according to the manufacturer's instructions to introduce the F184C mutation into the human Kv1.1 subunit cDNA. The nucleotide sequences of all linked subunits and throughout the joined segments were determined by automated sequencing. Oligonucleotides were obtained from EUROBIO (Milan, Italy). All channel subunits were subcloned into the oocyte's expression vector pBF, which provides 5′- and 3′-untranslated regions from the Xenopus β-globin gene flanking a polylinker containing multiple restriction sites. Capped mRNAs were synthesized in vitro using the SP6 mMESSAGE mMACHINE kit (Ambion).
Oocyte preparation and RNA injection.
Procedures involving Xenopus laevis and their care were conducted in conformity with institutional guidelines, which are in compliance with national (D.L. no. 116, G.U., Suppl. 40, 18 February 1992) and international [EEC Council Directive 86/609, OJ L 358,1, 12 December 1987; National Institutes of Health Guide for the Care and Use of Laboratory Animals (NIH Publ. No. 85-23, 1985); and "Guidelines for the Use of Animals in Biomedical Research" (Thromb Haemost 58: 1078–1084, 1987)] laws and policies.
The animals underwent no more than two surgical operations, with an interval of ≥3 wk between the procedures. X. laevis were anesthetized with an aerated solution containing 5 mM 3-aminobenzoic acid ethyl ester methansulfonate salt and 60 mM sodium bicarbonate (pH 7.3). The ovary was dissected, and the oocytes were digested in OR-2 solution containing collagenase A (Sigma; 0.5 U/ml). Stage V–VI oocytes were isolated and stored at 16°C in fresh ND96 medium (96 mM NaCl, 2 mM KCl, 1 mM MgCl2, 1.8 mM CaCl2, 5 mM HEPES, and 50 μg/ml gentamicin).
After 24 h, a nanoliter injector (WPI, Nanoject, Drummond, Broomall, PA) was used to microinject in vitro transcribed mRNAs into the oocytes, which were then incubated at 16°C. Typically, every oocyte was injected with 50 nl of a solution containing the relevant mRNA. The amount of mRNAs was quantified using a spectrophotometer and by ethidium bromide staining. Human Kvα1.4-1.1 and human Kvβ1 mRNAs were injected into the oocytes in a ratio of 1:20–1:40.
Electrophysiological recordings and data analysis were performed as previously described (14, 15). Briefly, two-electrode voltage-clamp recordings were obtained from Xenopus oocytes at ∼22°C 1–8 days after injection. An amplifier (GeneClamp 500, Axon Instruments) interfaced to a personal computer with an ITC-16 interface (Instrutech) was used. Microelectrodes were filled with 3 M KCl; resistance of the microelectrodes was 0.1–0.5 MΩ. The recording solution contained (in mM) 96 NaCl, 2 KCl, 1 MgCl2, 1.8 CaCl2, and 5 HEPES (pH 7.4). Currents were evoked by voltage commands from a holding potential of −80 mV. The recordings were filtered at 2 kHz and acquired at 5 kHz with Pulse software (HEKA Elektronik). Data analysis was performed using IGOR (Wavemetrics), PulseFit (HEKA Elektronik), and KaleidaGraph (Synergy Software) software. Leak and capacitative currents were subtracted using a P/4 protocol. An unpaired Student's test was used to determine the statistical significance. P < 0.05 was considered significant.
Functional expression of wild-type and F184C channels composed of Kv1.1, Kv1.4, and Kvβ1.1 subunits.
To investigate the effects of the F184C mutation on the biophysical properties of heteromeric channels and to assess their sensitivity to extracellular Zn2+, the human Kv1.4 subunit was concatenated with the Kv1.1 wild-type or Kv1.1F184C, as dimers, by means of a flexible glutamine linker. Two constructs were generated: Kv1.4-1.1 wild-type (WT) and Kv1.4-1.1F184C (F184C). The expression of these concatemers in Xenopus oocytes gave rise to functional delayed-rectifier currents, presumably arising from a homogeneous population of heterotetrameric channels composed of two subunits of each type. Figure 1A shows a representative current family recorded in two-electrode voltage-clamp configuration from a cell expressing WT channels. These currents were characterized by rapid N-type inactivation conferred by the Kv1.4 subunit, which possesses an inactivation particle at its NH2 terminus (Fig. 1, A and inset) (39, 43, 50). The dimeric channel was also coexpressed with the auxiliary subunit Kvβ1.1. As expected, the current family recorded from these cells showed that the human Kvβ1.1 subunit further accelerated the inactivation of the channel by providing four additional inactivation particles (Fig. 1, B and inset) (26, 38, 39, 47, 69). The current traces recorded from cells expressing the F184C channels showed altered kinetics of activation and inactivation in the presence and absence of Kvβ1.1 subunits (Fig. 1, C and D; see below).
F184C mutation increases Zn2+-induced inhibition of heteromeric channels.
To determine the Zn2+ sensitivity of WT and F184C channels with or without Kvβ1.1 subunits, macroscopic currents were recorded before and during the superfusion of a solution containing ZnCl2 at several concentrations into the recording chamber. The concentration-response curves for Zn2+-induced inhibition were calculated at 0 and +60 mV (Fig. 2). However, the open probabilities of WT and F184C channels, with or without Kvβ1.1 subunits, are maximal at +60 mV before and after the application of Zn2+, which shifted the V1/2 of channel activation only 3–10 mV to more depolarized potentials (not shown). This test potential also corresponds approximately to the peak of the action potential, and it allows us to obtain pathophysiologically relevant information. The Zn2+ concentration/inhibition data points for WT channels were best fitted by the sum of two Hill equations in the presence or absence of Kvβ1.1 subunits (Fig. 2). These results revealed, first, that heteromeric channels composed of Kv1.4-1.1 or Kv1.4-1.1/Kvβ1.1 subunits possess a high- and a low-affinity binding site for Zn2+. The low- and high-affinity equilibrium dissociation constants for Zn2+ binding (KZn) were calculated at 0 and 60 mV (Table 1). Unexpectedly, we observed that the auxiliary Kvβ1.1 subunits increased the sensitivity of WT channels to Zn2+-induced inhibition by decreasing the low-affinity KZn ∼2.7- and ∼3.8-fold at 0 and +60 mV, respectively (Table 1). The concentration-response relations were shifted leftward by the F184C mutation (Fig. 2). Overall, the F184C mutation significantly decreased the KZn values of the high- and low-affinity binding sites to Zn2+-induced current inhibition of Kv1.4-1.1 and Kv1.4-1.1/Kvβ1.1 channels (Table 1). Moreover, the low- and high-affinity KZn values of WT and F184C channels were markedly smaller at less depolarized potentials (Table 1) (24). Taken together, these findings imply that Zn2+ may regulate the transmission of signals by modulating the availability of heteromeric WT channels at distinct synapses. The availability of F184C channels is further reduced by Zn2+ because of their higher sensitivity for this metal ion.
Effects of Zn2+ on activation kinetics of F184C channels.
Little information is available regarding the dynamics of Zn2+ within the synapse. In particular, the mechanisms regulating the patterns of release, the peak concentrations, the rates of removal, and the distance that it disperses from its release site(s) are not well understood. Measurements obtained from imaging or use of chelating agents suggest that the peak concentrations of Zn2+ in the synapse may be transiently quite high, quickly dissipating to 10–30 μM (20, 35, 54, 58). Thus we used 10 μM Zn2+ for our study, inasmuch as this concentration is likely to be within a physiological range at mossy fiber synapses (61) and is comparable to the high-affinity KZn determined in vitro from our heteromeric constructs. In a previous study, we showed that the activation kinetics of homomeric F184C channels were markedly slowed by Zn2+ (14). To investigate the effects of Zn2+ on heteromeric channel activation, current traces were recorded at +60 mV from oocytes expressing WT and F184C channels before and after the superfusion of 10 μM Zn2+. The overlaid traces show that Zn2+ slows the kinetics of activation of F184C channels considerably, whereas those of WT channels were little affected by this ion (Fig. 3A). To quantify this phenomenon, a single-exponential function was fitted to the activation traces recorded at several test potentials for WT and F184C channels before and after application of 10 μM Zn2+. Plotting the activation time constants as a function of voltage revealed that Zn2+ increased the activation time constant of F184C channels nearly twofold at V1/2, whereas WT channel activation was much less affected by this ion (Fig. 3B, Table 2). The F184C mutation by itself confers slower kinetics of activation to heteromeric than to WT channels (Fig. 3B, inset; Table 2). The superimposed traces in Fig. 3A also suggest that the mutation itself and Zn2+ affect the time necessary to reach maximal channel activation. Thus the time to reach peak current was calculated as the time from zero current to its maximal amplitude and was plotted as a bar graph. This analysis confirms that the F184C mutation increased the time to maximal channel activation nearly threefold compared with WT channels (Fig. 3C). Furthermore, superfusion of 10 μM Zn2+ onto oocytes expressing F184C channels significantly increased their time to reach peak current (Fig. 3C). The coexpression of Kvβ1.1 subunits apparently accelerated the activation kinetics of WT channels (Fig. 3E, Table 2), and, again, the F184C mutation by itself slowed the channel opening kinetics (Fig. 3E, Table 2). The application of 10 μM Zn2+ markedly increased the time constants of activation (Fig. 3E, Table 2) and the time to reach peak current of F184C/Kvβ1.1 channels (Fig. 3F, Table 2).
Effects of Zn2+ on N-type inactivation and refractory period of F184C channels.
The Kv1.4-1.1 channels undergo fast N-type inactivation, which is accelerated by the ancillary subunits Kvβ1.1. Fast inactivation regulates the firing properties of neurons and their response to input stimuli. Thus the effects of the F184C mutation and Zn2+ on the biophysical properties of the channel were investigated. Figure 4, A–D, shows representative currents, evoked at +60 mV, before and after application of 10 μM Zn2+ for WT and F184C channels with or without the coexpression with Kvβ1.1 subunits. These superimposed traces suggest that the F184C mutation by itself and the superfusion of Zn2+ changed the amount and the rate of N-type inactivation induced by the Kv1.4 and Kvβ1.1 subunits. The amount of inactivation was then quantified as the ratio of final (at the end of the 1-s depolarizing pulse at +60 mV) to peak current amplitude. The amount of F184C current undergoing inactivation in the presence or absence of auxiliary subunits was significantly decreased compared with the amount of WT current (Fig. 4, E and F). The application of 10 μM Zn2+ reduced these amounts markedly (Fig. 4, E and F). To determine the inactivation rates, the decaying current traces for WT and F184C channels were fitted with a single-exponential function. The relevant time constants, plotted as a function of voltage, showed that the rate of inactivation was ∼1.5-fold slower for F184C than for WT channels at +60 mV (Fig. 4G, Table 2). The currents recorded from oocytes coexpressing the Kvβ1.1 subunit were best described with a sum of two exponentials. Again, the F184C mutation slowed the fast component of the time constants nearly twofold, which accounted for 85% of the current (Fig. 1F, Table 2). The superfusion of 10 μM Zn2+ markedly slowed the N-type inactivation rates of F184C channels in the absence and presence of the Kvβ1.1 subunit (Fig. 4, G and H, Table 2). By contrast, the effects on WT channels were much less pronounced (Fig. 4, G and H, Table 2).
To determine the refractory period of all channel types, a double-pulse protocol was used. Two test pulses to +60 mV were delivered for 200 ms from a holding potential of −80 mV and were separated by an interpulse interval of increasing duration. Figure 5, A and C, shows representative current traces of the recovery from inactivation for WT and F184C channels (alone or coexpressed with Kvβ1.1) before and after the superfusion of 10 μM Zn2+ into the recording chamber. To calculate the time constants of the recovery from N-type inactivation, the current amplitudes were normalized, plotted as a function of the interpulse duration, and fitted with a single-exponential function (Fig. 5, B and D). This analysis revealed that the recovery from inactivation was ∼1.4-fold slower for the F184C than for the WT channel (Fig. 5B, Table 2). In the presence of the Kvβ1.1 subunit, the recoveries from inactivation for WT and F184C channels were not significantly different (Fig. 5D, Table 2). Application of 10 μM Zn2+ markedly slowed the recovery of the F184C channel coexpressed with the Kvβ1.1 subunit (Fig. 5D, Table 2).
Effects of Zn2+ on voltage dependence of F184C channel inactivation.
The effects of the F184C mutation on the voltage dependence of heteromeric channel inactivation (steady-state inactivation) were assessed before and after application of 10 μM Zn2+. The amplitude of the outward current was measured during 100-ms depolarizing steps to +60 mV (Fig. 6, A–C) after 1-s prepulse potentials from −100 to +60 mV (in 5-mV increments). These currents were normalized, plotted as a function of prepulse potentials, and fitted with the Boltzmann function: I = 1/{1 + exp [(V − V1/2)/k]}, from which V1/2 and the slope factor k were calculated. This analysis showed that the F184C mutation shifted the voltage dependence of the steady-state inactivation of the channel ∼29 and ∼14 mV with and without Kvβ1.1 subunits, respectively (Fig. 6, B–D, Table 2), whereas application of 10 μM Zn2+ did not affect the inactivation voltage dependence of WT and F184C channels with or without Kvβ1.1 subunits (Fig. 6, B–D, Table 2).
In this study, we have demonstrated that distinct heteromeric channels have low- and high-affinity binding sites for Zn2+ and that the EA1 mutation F184C endows these channels with a higher sensitivity to Zn2+. Furthermore, we show that the F184C mutation will not only affect the function of the homomeric Kv1.1 channel, but it will also alter the biophysical properties of channels composed of the human Kv1.1, Kv1.4, and Kvβ1.1 subunits.
Structural and neurophysiological speculations.
The S1 segment plays a critical role in homo- and heteromultimerization of Kv1.x subunits and appears to stabilize subunit interactions (5, 41, 52). The F184 amino acid resides within the S1 segment. Here, we demonstrate that a phenylalanine-to-cysteine substitution at position 184 and Zn2+ slow the activation kinetics of the channel as well as the N-type inactivation kinetics and reduce the inactivating current amplitude. These results suggest that the F184C mutation and Zn2+ stabilize the closed-channel conformation, thus delaying channel opening. N-type inactivation results from the open channel block operated by the NH2 termini of the Kv1.4 and Kvβ1.1 subunits (47, 69). The activation and inactivation processes of Kv channels are intimately related. Therefore, it is not surprising that a slower time course of channel opening delays the onset of "ball-and-chain" block mediated by these subunits. On the contrary, an interruption of the normal time course of channel opening by N-type inactivation could account for the apparent faster activation kinetics of WT channels that we observed on coexpression of Kvβ1.1 subunits (18). The slower time course of F184C channel activation may possibly affect the time course of inactivation. Nevertheless, the functional characterization of Kv1.4-1.1I177N, Kv1.4-1.1E325D, and Kv1.4-1.1V408A channels shows that they undergo slower inactivation kinetics (unpublished results). These latter EA1 mutations either do not change or accelerate the activation kinetics of the channel (1, 16, 31). Therefore, it appears that several EA1 mutations slow N-type inactivation, regardless of their activation kinetics. Thus we cannot exclude the possibility that the F184C mutation and Zn2+ alter the affinity of the receptor site for the inactivation particle by allosteric modifications of the inner vestibule. Alternatively, they could change the overall structure of the nearby T1 domain, altering the predocking site for the inactivation particle before insertion into the inner vestibule (69). A similar mechanism has been suggested by Li and co-workers (34), who proposed that the EA1 mutation V408A, at an equivalent position of the Kv1.4 channel, modulates inactivation through membrane-spanning mechanisms involving S6 (34). Nevertheless, further studies are required to assess with certainty the molecular mechanisms underlying the altered biophysical properties of the channel caused by the mutation and by Zn2+.
Previous data showed that Zn2+ affected current amplitudes, kinetics, and voltage dependence of F184C channels also in the presence of external solutions containing higher concentrations of divalent cations such as Ca2+ and Mg2+ (14). Therefore, surface charge screening appears to be an unlikely mechanism underlying the Zn2+-induced effects on F184C channels. The concentration-response relation for the Zn2+-induced inhibition of the Kv1.4-1.1/Kvβ1.1 channel was well fitted with the sum of two Hill equations, suggesting the presence of two sites with which Zn2+ interacts. The molecular determinants of these binding sites are unknown. Recent studies of Kv1.5 channels have shown that Zn2+ may act on two distinct binding sites, one of which is located within the pore turret (32, 67, 68). It is possible that at least one Zn2+ binding site is also located within the pore turret of the Kv1.4-1.1 channel, since the amino acid sequence of this region is similar to that of the Kv1.5 channel. Unexpectedly, we also observed that the Kvβ1.1 subunits markedly decreased KZn for the low-affinity binding site. The mechanism underlying this effect is unknown. Nevertheless, allosteric modifications of the WT protein, induced by the association with the ancillary subunits, may possibly account for this phenomenon. Interestingly, the F184C mutation enhances the Zn2+ sensitivity of the low- and high-affinity sites. These results support the hypothesis that the F184C mutation strengthens the affinity for Zn2+ of sites already existing within the channel protein. Whether this occurs through allosteric changes caused by the mutation or by different mechanisms remains to be established.
The KZn values of the low- and high-affinity binding sites for Zn2+ on Kv1.4-1.1/Kvβ1.1 channels appear to be within the physiological range for peak Zn2+ concentrations at the synapse. These findings imply that Zn2+ may regulate the transmission of electrical signals by modulating the availability of heteromeric WT channels at synapses where Zn2+ is coreleased. The mossy fibers from the hippocampal CA3 area contain one of the highest concentrations of chelatable Zn2+ in the brain, which is assumed to be coreleased with glutamate. After release into the synaptic cleft and diffusion, hippocampal mossy fiber Zn2+ may modulate different pre- and postsynaptic mechanisms and may be taken up by Zn2+ transport systems or enter the postsynaptic neurons. Recently, it has been reported that Zn2+ released from mossy fibers activates presynaptic ATP-sensitive K+ channels (6, 45). These authors concluded that the activation of such a channel type may rapidly hyperpolarize the cell, reducing Ca2+ influx and glutamate release. On the contrary, our results suggest that Zn2+ facilitates synaptic transmission by depressing the activity of presynaptic delayed-rectifier K+ channels composed of Kv1.1, Kv1.4, and Kvβ1.1 subunits. Previous data have shown that Kv1.1, Kv1.4, and Kvβ1.1 subunits are the most likely molecular counterparts of the A-type current recorded from hippocampal mossy fiber boutons, which regulates the action potential duration and the frequency-dependent action potential broadening of the synaptic terminal (23). Here we have shown that Zn2+ reduces the availability of Kv1.4-1.1/Kvβ1.1 channels. These results support a model by which reduced delayed-rectifier K+ currents broaden presynaptic action potentials during high-frequency stimuli, leading to an enhancement of intracellular Ca2+ signals, glutamate release, and excitatory postsynaptic potential amplitudes.
Functional properties of heteromeric channels altered by the F184C mutation and implications for EA1.
Detailed biochemical studies indicate that Kv1.1, Kv1.4, and Kvβ1 subunits may also form heteromeric channels in the substantia nigra, globus pallidus, and cortical interneurons (48, 49). The results of the present study demonstrate that the F184C mutation will not only affect the function of the homomeric Kv1.1 channel, but it will also alter the biophysical properties of channels composed of the human Kv1.1, Kv1.4, and, Kvβ1.1 subunits. This evidence strongly suggests that the electrical properties of neurons within these brain structures, which coexpress these subunits, will be altered by the EA1 mutation F184C. The currents arising from the activation of Kv1.4-1.1/Kvβ1 channels possess A-type K+ current characteristics. A-type currents regulate the firing properties of neurons, their response to input stimuli, the propagation of axonal action potential, and their backpropagation in dendrites (13, 17, 28). Because of the localization of Kv1 channel complexes on axons and terminals of hippocampal synapses, it is expected that the action potential propagation and glutamate release might be modified by the mutation (44). Precisely how will the F184C mutation affect nerve cell properties? The functional characterization of WT and F184C channels, with and without Kvβ1.1 subunits, showed that the EA1 mutation causes opposite effects on the biophysical properties of the channel. On one hand, the F184C mutation downregulates the heteromeric channel function by slowing the activation rate and increasing the time to peak current amplitude. On the other hand, this mutation increases channel availability by decreasing the rate and the amount of current undergoing N-type inactivation and by shifting the voltage dependence of inactivation in the positive direction. The relative weight of these opposite effects has yet to be determined; therefore, it is difficult to predict specifically how they will affect the electrical properties of distinct neurons.
The functional characterization of the Zn2+ effects on F184C channels with or without Kvβ1.1 subunits showed that this ion 1) inhibited the F184C channels significantly more than the WT channels, 2) further slowed the activation rate of the channel, 3) further increased the time to reach peak current amplitude, 4) further decreased the rate and the amount of current undergoing N-type inactivation, and 5) slowed the repriming of the channel. First, these results demonstrate that the EA1 mutation F184C will not only sensitize the homomeric Kv1.1 channel to extracellular Zn2+, but it will also endow the heteromeric channels with a higher sensitivity to Zn2+ when the mutated subunit is associated with the human Kv1.4 and Kvβ1.1 subunits. Although Zn2+ also exerts some opposite effects, it appears that this metal ion mainly depresses F184C channel functions. Therefore, it is reasonable to hypothesize that the electrical properties of mossy fibers are likely to be altered by the combined effects of the F184C mutation and Zn2+. In particular, the reduced availability of Kv1.4-1.1F184C/Kvβ1.1 channels, because of their increased sensitivity to Zn2+, may further increase the excitability and the frequency-dependent action potential broadening of the synaptic terminals, leading to an enhancement of intracellular Ca2+ signals, glutamate release, and excitatory postsynaptic potential amplitudes. Interestingly, EA1 patients report cognitive symptoms during an attack. An alteration of the transmission of the electrical signals within the hippocampus may possibly underlie this symptom (42). Moreover, epileptiform brain activity has also been associated with intracranial administration of Zn2+ salts, and changes in Zn2+ modulation of GABA receptors have been implicated in the etiology of epilepsy. Interestingly, patients bearing the F184C mutation have shown enhanced seizure susceptibility in addition to typical EA1 symptoms (60). Whether Zn2+ plays a role in triggering epilepsy-like symptoms in this kindred remains an intriguing hypothesis.
In conclusion, one major finding of this study is that heteromeric channels composed of the human Kv1.1 and Kv1.4 subunits have low- and high-affinity binding sites for Zn2+, which are likely to regulate channel availability at distinct presynaptic membranes. We also showed that the EA1 mutation F184C alters the biophysical properties and the Zn2+-induced modulation of such heteromeric channels with or without Kvβ1.1 subunits. These molecular mechanisms of EA1 are likely to exacerbate the symptoms in patients harboring the F184C mutations by altering the transmission of signals within nerve cells expressing this heteromeric channel type. Direct evidence of distinct electrical events modulated by Zn2+ and of overt behavioral changes caused by the F184C mutation shall be provided by the in vitro and in vivo analysis of wild-type and knock-in mice harboring such point mutation (2, 37).
This work was financially supported by Telethon-Italy Grant GGP030159, MIUR-COFIN 2005, and Compagnia di San Paolo (Turin, Italy). P. Imbrici is the recipient of a Ph.D. fellowship from Compagnia di San Paolo.
We thank Ezio Mezzasoma for technical assistance and Domenico Bambagioni for art work. We are grateful to the Italian Red Cross (Women's Section of Perugia) for the donation of scientific equipment to the Section of Human Physiology for this research.
Present address of A. Cusimano: Istituto di Biomedicina e Immunologia Molecolare, CNR, Via Ugo La Malfa 153, 90146 Palermo, Italy.
The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.
- Copyright © 2007 the American Physiological Society


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This thread could be used for your own requests, a general Make-over thread.
Ahhh...went ahead and took the plunge, leaving the thread up though for others to post such requests (if mod feels like its a waste, delete away). My choice went with thinner, darker, face change, typical frosted thief look:
Edited by TGIFrisbie, 27 November 2012 - 05:30 PM.

subset: commoncrawl


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Jim Malloy has medical problem detected during student training exercise
(WJLA) - Jim Malloy, a Charlottesville retiree, was told to pretend he had an abdominal aortic aneurysm by University of Virginia doctors, not actually have one.
When he did though, he was luckily in the right place to have his life saved.
Malloy and his wife, Louise, regularly would pretend to suffer from a wide variety of maladies in the name of training aspiring doctors at the University of Virginia School of Medicine. In March, Jim was asked to pretend to suffer from an AAA. It's a condition where a section of a person's lower aorta balloons.
That's when, according to the university, UVA medical student Ryan Jones noticed the symptoms of an actual aneurysm. Several months later, Malloy checked into the hospital, had a stent implanted and is now doing just fine.
"It feels great to know that I made a difference," Jones said.


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The Department of Surgery is dedicated to the education of medical students. Our department provides a unique learning experience for third- and fourth-year medical students.
Visit the UW SMPH Health Education site for additional information.
Opportunities are also available for medical students at other institutions to complete clinical rotations as a visiting student. The Department of Surgery is also pleased to offer the Underrepresented Minority Visiting Elective Scholarship Program.


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Get DVD Talk Feeds
For those not familiar with The Andy Milonakis Show, in a nutshell it's a sketch comedy show about Andy (a thirty year old who thanks to a growth hormone problem looks like he's in his early teens) who lives in an apartment next to an old Jewish guy named Larry and a weird black dude with a speech impediment named Ralphie. He periodically receives visits from an old woman named Rivka and a nameless old black man wearing a flannel shirt, smoking a cigarette and holding a dead fish (this guy is both completely hilarious and utterly terrifying at the same time – like much of this show!). Andy has little misadventures, many of which involve his friends and neighbors, and he also tends to head out onto the streets to talk to other New Yorkers.
What the show provides is a completely absurd mixture of skits and interviews done guerilla style with various random citizens of the Big Apple with the odd animated cartoon thrown in to the mix just for kicks. None of it makes a lick of sense and it's all just really, really, really weird. It's also damn funny.
First and foremost when thinking about what makes the show work is Andy himself. He's a brat, or at least he plays one really well, who often gives in to his selfish needs and mistreats his friends. He's a prankster with a love for messing with home delivery services and busting out primitive freestyle raps. He's a fat kid in a red shirt who'll eat strange things for your amusement and who periodically flies around on his pet dog who he seems to both adore and fear. Andy is the center of the strange universe that this show exists in, and everything seems to revolve around him.
As funny as Milonakis can be, the show would not be the same without the supporting cast. Ralphie may or may not be mentally retarded, we don't really know, while Larry continues to amass a collection of can openers that he's been borrowing from Andy and not giving back for God only knows how long. Rivka is just so flat out strange looking with her lip curling up and her thick Coke-bottle glasses that you can't help but be amused by her very presence and the crazy guy with the fish shows up at such random and inopportune times that he is always good for a laugh even if you won't really know what exactly it is you're laughing at.
The second season of The Andy Milonakis Show contains eight half hour episodes that are presented here uncut and like the first season, these episodes sometimes contain some fun guest stars. Appearing in the second seasons are Seth Green, Jimmy Kimmel, snowboarder Shawn White, Julez Santana and a few other notables. The guest stars are always involved in the skits – Shawn White goes snowboarding with Andy while Kimmel explains how 'he made' him at which point we see Andy being created by a bearded Jimmy in a funny Pinnochio parody that eventually goes horribly wrong much to the dismay of a convenience store owner.
The comedy in the show won't appeal to everyone. Humor is, after all, extremely subjective and so much of this material is just so completely out there that it isn't really the most accessible sketch comedy show in the world. It makes oddities like The Kids In The Hall seem positively normal by comparison but it is definitely hilarious stuff if you're to appreciate the unfiltered absurdity of it all. It's low brow at its finest but at the same time it can also be rather clever which makes it unique, stupid, irritating and wonderful all at the same time.
The 1.33.1 fullframe transfer on this set maintain the show's original aspect ratio and for the most part they all look pretty good on DVD. Some shimmering is present in a few scenes but there's no need to worry about edge enhancement or compression artifacts. Color reproduction is fine and there's as much detail in the image as you'd want to see. For a show that's shot on DV things look quite strong here and there aren't any issues with print damage or debris, the picture is clean and stable throughout playback.
The English language Dolby Digital 2.0 Stereo track on the set isn't going to make you want to make this your new demo DVD but it sounds as good as it needs to. Dialogue is clean and clear and while sometimes the outdoor bits have a bit of background noise, you won't ever have a problem understanding anyone. The music is punchy and strong as are the effects but the levels are properly balanced and nothing ever gets buried. There are no subtitles, alternate language dubs or closed captions provided for this set.
Each of the episodes in the set has an optional commentary track with Andy, who is joined periodically by a few of the other performers including Ralphie, Larry, Rivka and Billy. These tracks aren't in the least bit informative, in fact, they're pretty much just nonsense, much like the show. That being said, they can be pretty funny. Much of what we hear is observational humor, remarks about what's happening on the screen rather than how it happened or how it was put together. Don't expect much in the way of serious trivia or an honest discussion of how the show is put together, instead we get a lot of ribbing and riffing on one another and the kind of general goofing off that one would expect from Andy and his crew.
On the second disc you'll find roughly ten minutes worth of unaired skits. You can watch each on individually or if you select the first one and just let it play it defaults to a 'play all' function. Some of this material is pretty funny and a lot of it is just as good as the skits that made it into the broadcast episodes so it's nice to seem them included here as they are definitely good for a few laughs. There are also a few minutes of outtakes in here and we get to check out how some of the skits and ideas used in the show can backfire which again is quite amusing. Also included on the second disc is the music video for the second season theme song. Each disc has animated menus and episode/chapter selection options.
If you're into absurdist sketch comedy and nutty man-on-the-street impromptu interviews then you should give The Andy Milonakis Show a shot. It's as wacky as wacky can get but it's just stupid enough to work especially in episodic doses. The Complete Second Season looks and sounds good and it's got some fun supplements on top of the episodes earning this set a solid recommendation.
Ian lives in NYC with his wife where he writes for DVD Talk, runs Rock! Shock! Pop!. He likes NYC a lot, even if it is expensive and loud.


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Aculco rentals, vacation homes direct from the owners
You will find many studios, apartments, houses and villas for your vacations in Aculco. Contact the owners directly on the IHA site and enjoy the best rental rates, promotions and last minute deals that meet your budget. Visit the photo gallery on private rentals for a break with family or friends. All vacation listings on IHA are verified, plan your trip to Aculco with absolute confidence!
6 bedrooms House for rent near Aculco from 16 to > 30 people
Tequisquiapan rentals, at 30mi. from Aculco - State of Querétaro - Central Mexico
direct by owners
35 requests already received
Advert 70709PDF
Show map
Quinta Cobos Country House > 0.25 acre in a private property. panoramic, mountain, swimming pool, garden/park viewHouse Rental in Tequisquiapan #70709 >Swimming poolParkingPlaypen
Interior facilities
- Sleeping den
- 11 Bathroom
- 6 Shower room
- 6 Toilets
- Living room 1,600 Sq.ft.
- Sitting room 970 Sq.ft.
- Countryside vacation
- Ideal for large families and group
- Kids welcome
- Setting : Enchanting, Tranquil
- River at <165'
- Meublés de Tourisme : 3 stars
- Golf, Festival, Spa, Horse riding, Fishing
View the advert


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Making the simple complicated is commonplace; making the complicated simple, awesomely simple, that's creativity
We specialise in designing state of the art websites for individuals and businesses worldwide. Our team is dedicated to help you get your business online and make it prosper. The websites we create are responsive and well attuned to the needs of our customers.
They are designed in an attractive and appealing way that makes it easy for your readers to get engaged with your content, services and products. We offer fully responsive user interfaces that are powerful and flexible enough to work on any type of mobile device or desktop while retaining the beauty and functionality of the original. Whether you own a website that you would like to see expanded, or you plan on being the owner of a brand new website, we can help you realize your ambition. Contact us now and see for yourself, you and your website's users will be glad you did!
We make websites that don't just look good, they work well, too. Unlike some digital agencies, we're not focused solely on creating works of art, we're passionate about user experience and conversion rate optimisation, too.
There is no point in reinventing the wheel and even less point in paying for it, thats why we use tried and tested open source platforms such as WordPress and Joomla to keep you costs down and your functionality high.
- We love what we do
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If you are looking for an indication of the breadth of our experience, then please visit our portfolio page to explore our website design, branding, UX-UI and CMS services. Or if you'd like to know what our clients say about working with us, then please visit our testimonials page.
Fantastic work - delivered on time and went to the extra mile - Highly recommended.Mike LowndesFounder
Couldn't have asked for a better product! Perfect in every way, thank you very much for not just meeting our expectations, but exceeding them!!!!Simona THead of Marketing
Working with Rob has been a pleasure on so many levels. He is imaginative, thoughtful, knowledgable and a great communicator. The site he's produced is beautiful and I'm delighted.Eliott ScottOwner
Rob did an amazing job - easy to work with, stays on schedule, and delivered a great final product!Jaydon RobinsonCo-founder
We engaged Rob to design the homepage and inner page of our website. He worked hard to complete the design of our website ontime and onbudget, and worked with us thru various revisions and iterations of the webdesign. He takes on board feedback well, and has good web design skills, and in addition he is responsive and prompt in the delivery of his work.David COwner
Great to work with. Worked within budget and time frame. The results were very skilled. He interpreted my objectives perfectly.Carl LOwner
We'll bring your vision to life.
If you're reviewing the effectiveness of your existing web presence, or thinking about starting something new...


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Your password reset has been submitted to the address provided.
Design plus words, letters, and/or numbers
Medical services, namely, providing medical information including clinical study results to doctors and patients relating to diseases and disorders of the cardiovascular system and metabolism
Business in Basel, CH


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New Rudy Currence Songs, Mixtapes & Videos
Rudy Currence - Royal Blue [Stream]
It's been a year since we last heard from Rudy Currence (Blackberry Molasses is still in rotation in our office), but today the Rock Hill, South Carolina native returns with a new single in hand. Royal Blue, produced by... Read More
Rudy Currence - Blackberry Molasses [Stream & Download]
Heartache and pain will be part of even the happiest life. Fortunately, we have music to help us through the toughest of times. If you're feeling down, Rudy Currence's latest single, Blackberry Molasses, may be... Read More
Rudy Currence - Reinvention [Stream]
It's hard to believe we're already ten days into the New Year, isn't it? Hopefully the prospect of new beginnings hasn't worn off just yet. Let's face it, even if you don't believe in... Read More
Rudy Currence - The O.T.H.E.R. [Stream & Download]
Previously featured on summer 2010's Anymore, Dirty Southern crooner Rudy Currence makes his return to our pages with the latest single off his forthcoming LP. The O.T.H.E.R. finds the DTP crooner showcazsing his... Read More
Rudy Currence - Anymore [Stream]
Impassioned, often crazily-elaborate metaphors have been a staple of romantic discourse since… well, at least since the Song of Solomon (y'know, that sexy part of the Old Testament that fundamentalists do their... Read More


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Google took the consumer electronics world in July by storm with the announcement of the [easyazon-link asin="B00DR0PDNE" locale="us"]Google Chromecast HDMI Streaming Media Player[/easyazon-link], a digital media dongle available at the unbelievably low price point of $35.
The arrival of the Chromecast was a big surprise for several reasons. I keep on top of all the tech rumor sites every day, and I hadn't heard so much as a rumor that Google was working on a media dongle, much less one for $35. There's also Google's very checkered record when it comes to hardware releases, which included the "Nexus Q," another media streaming device introduced in the summer of 2012 that was ultimately cancelled before release.
But the Chromecast was something different: A super-cheap streaming solution, controllable from a smartphone or tablet, and a product that was released the day it was announced. In addition, it offered "browser-casting" from the Google Chrome browser on a computer. That, and the low price point, led to the Chromecast's instant popularity.
One morning the week it came out, I called several local Best Buys. Most had none available, except one had nine of them, from a new shipment. I went at lunch that day- two hours after the phone call- and they were all long gone.
After I ordered it online and waited about a month for the shipment, I've now been using the Chromecast for about three months, and the device has its pluses and minuses. But with the exception of its inclusion of YouTube- which is owned by Google, of course- there's not a lot about the Chromecast that I prefer over my trusted Roku LT. I have kept the Roku in my living room, with the Chromecast finding a home attached to my living room TV.
Here's how it works: The Chromecast is a tiny dongle that attaches to a TV's HDMI port- it's so tiny, in fact, that since the day I attached it to my TV I haven't even looked at it. It connects to your home Wi-Fi network, and can be controlled from an iPhone or iPad. There's a Chromecast app, but I've found I control the device mostly from within the Netflix and YouTube apps.
So what's good about the Chromecast? It's easy to use and the interface is excellent. It's not buggy; it works, every time I try to use it. The small size and small price are to recommend too; it's cheaper than the least expensive Roku device and most other competing streaming boxes, Apple TV included. When my kids roll into our room at 6 a.m. and demand a cartoon, dialing up a Daniel Tiger's Neighborhood episode on Netflix via the Chromecast is much easier than navigating those awful cable on-demand menus.
The drawbacks, though, are plentiful, starting with the woeful collection of apps. The Roku, for instance, offers access to hundreds of apps. Chromecast launched with only Netflix, Google Play Movies, TV and Music and YouTube, and has since added Hulu Plus and Pandora.
Sure, if the brief era of Smart TV has taught us anything, it's that there are only a handful of TV apps that people actually use. But the Chromecast omits Amazon, HBO Go, Facebook, Twitter and Spotify, as well as every major sports app. It also doesn't allow streaming of native content.
So overall, do I recommend the Chromecast? If Netflix and YouTube are the only streaming apps you care about- and yes, for many people, that's the case- than it's a good gateway for streaming. It's also perfectly fine for a kitchen or bedroom TV. But if you're interested in streaming everything there is to stream, and have no other options, go for the Roku instead.
Google Chromecast streaming dongle


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Legal Question in Wills and Trusts in Florida
How do you find an attorney's paperwork who did a will, now he is out of business?
Asked on 7/22/13, 10:55 am
1 Answer from Attorneys
Barry Stein De Cardenas, Freixas, Stein & Zachary
Each Florida attorney is supposed to name another attorney to receive and maintain his active files if something happens or he goes out of business. This does not mean that information with regard to a will will be in those files unless he maintained the original. Check with The Florida Bar to see if the attorney named the other attorney to hold his files.
Answered on 7/22/13, 1:48 pm


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I enjoy being a board member at the Empowerment Through Arts Foundation and working by our president's side. Monica is a goal-oriented professional with excellent interpersonal skills that allow her to liaise efficiently with stakeholders at all business levels. She has the ability to motivate and inspire those around her with her enthusiasm, creativity, high energy and engaging personality. Monica eagerly accepts new challenges and finds innovative ways of meeting them. As a trustee she does not think outside of the box because she does not even know that the box exists. That is an important trait for a leader to have when you are examining an organization and how it functions. Monica has demonstrated the ability to create solutions that fit the situation rather than embrace a one-size fits all mentality. She's resourceful and is a true administrator, fostering leadership qualities in others. Her leadership style is smart: allowing people to grow in their roles, welcoming new ideas, and providing direction with encouragement, earning well due respect from staff and volunteers in our campaign. I highly recommend her as a consummate professional with whom it is always a pleasure to work.
Review from Guidestar


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Currently 1.00/512345 Rating: 1.0/5 (Total votes: 1)
Rating: 1.0/5 (Total votes: 1)
1. Manage notes both text and image (single image per note)
2. Lao Keyboard layout
3. Search notes time, text wise.
4. Share to all popular social networks like Facebook, Whatspp, Twitter Or you can email, sms to anyone in Khmer.
5. Texts are autosaved as soon as you write.
The app should help all Lao users in Thailand, Laos, Cambodia and Lao native speakers all over the world .
» show more


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Recent Posts
Category Archives: North Carolina
Frankie Silver Murders Husband in 1831
Frances "Frankie" Silver was born in 1815. She was hung in Morganton North Carolina on July 12, 1833 for killing and dimembering her husband, Charlie Silver. She was only 18 years old at the time of her death. Frankie married … Continue reading
Posted in North Carolina Tagged 1833, Burke, charlie, frances, frankie, hanging, morganton, murder, north carolina, silver 14 Comments
Lawson Family Murders
Charles Davis Lawson was a tobacco farmer from Stokes County North Carolina. He owned a farm on Brook Cove Road near Germantown. He lived there with his wife, Fannie (37), and their seven children: Marie (17) Arthur (16) Carrie (12) … Continue reading
Posted in North Carolina Tagged 1929, charles, charlie, christmas, family, fannie, horror, killing, lawson, massacre, murder, north carolina, stokes 17 Comments


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Child abuse prevention and awareness walk scheduled in Chamberlain
CHAMBERLAIN -- First Child Abuse Prevention and Awareness Walk in honor of Mason Naser will be held at noon on Saturday at American Creek Campground in Chamberlain.
Raffle tickets will be sold for a star quilt and beaded earrings. Justice For Mason T-shirts will be available for $15 in adult sizes small to XL. After the walk, winners of the quilt and earrings will be announced.
Vegetable soup, fry bread, wojapi, hot dogs and chips will be available after the walk at the American Creek Campground shelter. Money from the raffle and T-shirt sales will go toward an Easter egg hunt in Lower Brule and sponsorship of Tiny Tots at the Lower Brule powwow. Information will be presented during lunch about the impact child abuse has on families and victims.


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Artist description:
I love to draw some RMD things, like this one. I hope you like it. Thank You!
Canvas Draw Created Jul 26, 2011 640 x 460px Unless noted Copyright 2011 Yuki LEe.
We encourage you to submit a comment and let the artist know what you think of this drawing. Please log in or join to post a comment.
Post a comment
yeah... i didn't have my snacks for the 2nd time...
Ugh.... Are you Hungry,Saori? O.o
No It's only a drawing!!!
Hey Sis,Did You Cooked It Or Alfred Cooked It? O.o


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Use the tools below to refine your search by only displaying reviews with a certain number of star ratings or to only show reviews from a certain time period. Eg click on '1 star' to just display the reviews we have which received a 1 star rating click or 'Within the last month' to display just reviews posted over the last month.
As rated by our community of reviewers
" Catering at its Best " Read More
Written by Dawniey
" Outstanding food and service! " Read More
Written by Milliham
" Very rude woman " Read More
" Fantastic food & Staff " Read More
" Ruined Our Wedding " Read More


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Listing Courtesy of Ross Reed of Coldwell Banker Anchor R.E.
Listing information
- Beds: 4
- Stories: 0
- Foreclosure: No
- Lot Area: 0.12
- Association Fee: $210
- Subdivision: S30 - St. Joseph School
- Style: Condominium
- Baths: 5 (Full: 3 3/4: 0 1/2: 2 Other: 0)
- Parking Type Description: None 0 cars
- Total Square Footage: 2,160 ($203.66 per sq. foot)
- Lot Size: 23.79x163, 10x91, 113.24
- County: Berrien
- Year Built: 2004
- Status: Active
- New Construction: No
- Above Ground Sq. Feet: 2,160
- Estimated Annual Taxes: $10,454
- School District: St. Joseph
Access Feat: Accessibility Features No
Additional Items All Window Treatments, Ceiling Fans, Garage Door Opener, Wood Floor
Air Conditioning Central
Appliances Built in Oven, Cook Top, Dishwasher, Dryer, Freezer, Refrigerator, Washer
Assoc. Amenities Club House, Meeting Room, Pets Allowed, Pool - Outdoor, Pool-Indoor
Assoc. Fee Incl. Lawn/Yard Care, Snow Removal
Association Info.: Assoc. Fee Payable Monthly
Basement No
Design Contemporary
Driveway Paved
Exterior Features Deck(s)
Exterior Material Vinyl
Fireplace Living, Yes
Garage Type 2
Garage: Garage Yes
Heat Source Natural Gas
Heat Type Forced Air
Kitchen Features Snack Bar
Landscape Shrubs/Hedges, Underground Sprinkler
Lot Description Flag Lot
Manufactured: Manufactured Y/N No
Mineral Rights Unknown
Other Association Info.: Approx. Assoc Fee - 210.00, Lower Rooms: Lower Half Baths - 0, Lower Rooms: Lower Bedrooms - 0, Lower Rooms: Lower Finished SQFT - 0.00, Lower Rooms: Lower Full Baths - 0, Lower Rooms: Total Lower SQFT - 0.00, Main Rooms: Main Bedrooms - 2, Main Rooms: Main Full Baths - 2, Main Rooms: Main Half Baths - 1, Main Rooms: Main Level SQFT - 1260.00, Road Frontage - 23.79, Upper Rooms: Upper Bedrooms - 2, Upper Rooms: Upper Full Baths - 1, Upper Rooms: Upper Half Baths - 1, Upper Rooms: Upper Level Square Feet - 900.00, Waterfront Options: Water Frontage - 91.93
Pets Y/N Yes
Pool Outdoor/Inground
Property Sub-Type Condominium
Roofing Comp Shingle
Sale Conditions None
Street Type Paved
Substructure Crawl Space
Terms Available Cash/Conventional
Util Avail at Street Broadband, Cable, Electric, Natural Gas, Public Sewer, Public Water, Telephone Line
Utilities / Sewer Public
Utilities Attached Cable, Natural Gas, Telephone Line
Water Fea. Amenities All Sports, Deeded Boat Lot, Private Frontage
Water Heater Natural Gas
Water Type Public, River
Waterfront Options: Water Access Y/N Yes
Windows Low E
Bedroom 2 U 25x11
Master Bathroom U 13x9
Master Bedroom U 13x22
Bedroom 3 M 12x10
Bedroom 4 M 9x10
Kitchen M 17x13
Living Room M 23x17
Listing Courtesy of Ross Reed of Coldwell Banker Anchor R.E.


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Fitzpatrick sworn in as Rock County circuit court judge
Governor Jim Doyle has high praise for Rock County's newest circuit court judge. Doyle says when Michael Fitzpatrick interviewed for the Branch 3 appointment, he'd built up his legal practice, was highly regarded in the bar, and was at a point in his career where he could take the cases he wanted and train the younger lawyers to take the others. Doyle asked Fitzpatrick why he'd want to move to the bench. Fitzpatrick told Doyle during the interview about public service and dispensing justice.
Fitzpatrick says as he works in Courtroom A, some of the cases seem to be similar to others, but those cases aren't similar to those people who are in the courtroom. He says that is the most important thing going on in their life right then, and it's important to remember it's not routine for them.
Fitzpatrick was sworn in Thursday afternoon. He began his duties this week.
Scroll down to hear Doyle's comments from the investiture.
Last updated: 10:15 pm Thursday, December 13, 2012


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Dan Jordan, Spencer Larson and Chris Laughlin achieved the honour during the Consumer Electronics show at the Las Vegas Convention Center in Nevada.
The trio, all from Las Vegas, watched programing from two TiVo Roamio digital video recorders that had been broadcast and recorded ahead of the attempt
They were allowed breaks of five minutes per hour to use the bathroom or nap, in accordance with Guinness World Records rules for the category.
While allowed to save the five-minute breaks, the longest anyone slept during the 87 hours at any one time was 80 minutes.
They underwent regular medical checkups throughout and were allowed to change channels and eat and drink while they watched, but were not allowed to read or talk on the phone at the same time.
For completing the marathon, the three shared a prize of $5,000 (£3,038) and received a package including a TV, TiVo Roamio with product lifetime service, iPad, Sonos system, gamer chair and a year's Netflix subscription.
Mr Jordan told guinnessworldrecords.com: "When we arrived on Monday, we had no idea what we were in for. It's been a long week, but it's been fun."
The attempt broke a record set in 2012 by Carin Shreeves and Jeremiah Franco, which saw the pair watch episodes of The Simpsons for more than 86 hours.
:: Watch Sky News live on television, on Sky channel 501, Virgin Media channel 602, Freeview channel 82 and Freesat channel 202.


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Housing authority upgrades drafty windows
Wed, 22 Jan 2014 03:31:42 GMT —
The Lee County Housing Authority met Tuesday to go over the budget for the new year.
The budget is set around $480,000.
Lee County officials will soon start using funds to replace all the windows in one of their apartment buildings.
"It's Academy Homes. It's right off 5th Avenue in Academy and we'll be replacing the windows there with a contract of about $46,000," said Dan McCarthy, Executive Director for the Lee County Housing Authority.
McCarthy says they are also continuing to work on their rental assistance program.


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Healthcare: Cash Comes In, Jobs Go Out
By Kristen B. Frasch
Saturday, June 16, 2012
A survey by ADP shows companies of all sizes are choosing to outsource benefits administration because of the ever-increasing compliance complexity of, not just healthcare reform, but other benefits-related acts as well.
In addition, Kaiser Family Foundation finds that employers could be receiving millions in health-insurance rebates from heath-insurance companies that spent more on administrative and marketing expenses and profits than is allowed under the new law.
The report estimates rebates for large companies could reach more than $541 million.
Copyright 2017© LRP Publications


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We're The 'B' Team, oppurrating from a secret bunker somewhere in middle England ~ If mew have a purroblem, if no-purrdy else can help, and if mew find can us, then purrhaps mew can hire The 'B' Team. We also share a wide array of entertaining posts to amews mew each week, so drop by often to catch up on all the fun!
Tuesday, 3 May 2016
Tuesday Tails ~ Pirate Q&A ~The Secrets of The Voyages of Captain Basil Revealed ~ Part VIII & Part IX
7 comments:
guys.....a total lee awesum post two day & manee thanx for answerin everee onez questshunz.....ya noe, we never trusted that amsted dood either, like who wood with a name spelled bak werdz that iz detsma...ReplyDelete
now speekin oh weaponz....ewe noe we haz pie ratz connectionz over at thiz end oh de globe; sew if ya ever knead sum......just sayin with out....sayin
and speekin oh klingonz.....we bet horice knowz that hole transporter rooms stuff like frum star trek N thatz how ya get bak home...that ore thoz red shooz like that dorothy ladee wore !!!
we will take another round o three number 5's pleez....thanx !!♥♥♥Replies
Detsma!!!!!!!!!!!!!!!!!! MOL MOL MOL ROFMOL seriously mew guys crack us up so much, we love your comments!Delete
It's always good to have connections, and if we effur need some pirate dudes we'll let mew know - smoke signal ok?
Horice knows so much more than he's been letting on... but that's ok, I'm sure he'll impart knowledge when he's ready :D
And of course mew can have 3 more No: 5's and I'll throw in a few of the new recipes too - I'll dispatch them today via robot carrier pigeon.
Purrs Basil xox
You really outdid yourselves with answering the questions. I think it took up more lines than the actual story. I think you should get the series published as a book.ReplyDeleteReplies
MOL guys I think mew could be right! Do mew think it would translate ok in book form? Purrs Basil xoxDelete
I am really enjoying reding this!! PS Tell Mu your interview goes live tonight our time!!! Maybe when you wake up......ReplyDeleteReplies
Hi guys, we're so purrleased that mew like our Pirate Q&A and thanks fur letting us know about the interview, we've got a post published and are promo-ing as we type this! Thanks mew so much, purrs Basil & Co xoxDelete
and we can't spell either - reading.....ReplyDelete
*Waves Paw* we love comments and do purr extra loud when mew leave one, and we do try to reply to effurypurrdy and visit mew too! ❤❤❤


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Artist description:
i have a crush i hope she is lookin. guess who this crush is and i'll tell everyone who it is. just give a guess. if the right person ask ill. tell them.
Flash Draw Created Aug 16, 2007 470 x 338px Unless noted Copyright 2007 grim18.
We encourage you to submit a comment and let the artist know what you think of this drawing. Please log in or join to post a comment.
Post a comment
no.. haha it's ME! kidding kidding... good luck with ur crush!
this one is so cool
sarah?plz tell or is it messisa
Ehm.. I think it is me! xD :p


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Caught in the Sandwich Generation? Get Some Tax Relief
The term "Sandwich Generation" refers to adults in their 30s to 50s with children at home and responsibilities to care for aging parents. As a tax professional and a "Squashee" -- I have a daughter in Grade 7 and an elderly father living in my home -- I've got a unique perspective on the tax implications of this juggling act.


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Dr. Thomas Frieden, director of the Centers for Disease Control in Atlanta, talks to Charlie Rose and Gayle King about a new test that could make it easier to diagnose food poisoning. Some people are expressing concern, however, because the test could make it harder to track the source of future contaminations.
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Honeywell features an unrivaled selection of premium Door Chimes. Incredibly diverse by design, Honeywell has expanded its catalogue to include both intuitive and traditional Door Chimes. The Honeywell RCWL330A100/N P4-Premium Portable Wireless Door Chime and Push Button is a Wireless option that doubles as an extra defense in security. Obtain new levels in peace of mind with the ability to place Add-on push buttons and motion detectors that work in conjunction with your door chime. More traditional options such as the Honeywell RPW301A1009/A Wired Surface Mount Door Chime Push Button are incredibly easy to install Surface Mounted Door Chimes that feature all of the durability and performance one would associate with a superior Honeywell product. Regardless of whether you're shopping for a more traditional and Decorative Piece or a Wireless option that features security options, Honeywell has the appropriate Door Chime for you.
Let Honeywell door chimes help you show off your individual style! You'll be sure to impress your guests with items such as the Honeywell RCW3504N1001/N Decor Wired Door Chime Decor chime with its sleek wooden and glass look and multiple door chime options, or the RCWL3505A1005/N Decor Customizable Wood Wireless Chime, paint it, stain it, wallpaper it, make it your own!
Tired of having to replace your doorbell from UV damage or poor quality? Honeywell offers a wide variety of dependable and long-lasting surface mount doorbells to fit your needs. UV resistant doorbells such as the RPW210A1002/A Wired Surface Mount Illuminated Push Button Door Chime or RPW111A1002/A Wired Surface Mount Illuminated Push Button Door Chime are great options for withstanding constant UV exposure. Or if you're looking for something with a little character the stainless steel Honeywell RPW204A1005/A Wired Surface Mount Illuminated Push Button Door Chime, Stainless Steel Finish or solid brass RPW316A1007/A Illuminated Recessed Door Chime Push Button can help bring new life to your entrance!
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Clear Image Radio Interview, Sunday Feb. 17th!
by Eric Walter on January 28th, 2008
Blaine Duncan and Eric Walter will be guests on Clear Image Radio "Real Paranormal Talk" on February 17, 2008 at 10PM EST.
Be sure to tune in for exciting updates from AmityvilleFiles.com and call in to the show to voice your opinions on America's most famous haunted house controversy!
To listen live, visit http://clearimageradio.com/
AmityvilleFiles.com on X Zone Radio! 1/2/08
by Blaine Duncan on January 1st, 2008
Eric Walter and Blaine Duncan will be guests of X Zone Radio with Rob McConnell January 2, 2008 at 9PM PST. Be sure to tune in for exciting news from AmityvilleFiles.com, and listen to Eric and Blaine discuss the controversy surrounding the Amityville story!
To listen live, visit http://www.xzone-radio.com


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November 9, 2004
v No. 249226
Oakland Circuit Court
LC No. 2002-040915-CZ
Before: Murray, P.J., and Sawyer and Smolenski, JJ.
Plaintiff appeals as of right the order granting defendant?s motion for summary
disposition in this age discrimination and retaliation action under the Civil Rights Act (CRA),
MCL 37.2101 et seq. We affirm. This appeal is being decided without oral argument pursuant
to MCR 7.214(E).
A motion under MCR 2.116(C)(10) tests the factual sufficiency of the complaint. In
evaluating the motion, the trial court considers affidavits, pleadings, depositions, admissions and
other evidence submitted by the parties in a light most favorable to the party opposing the
motion. Where the proffered evidence fails to establish a genuine issue of material fact, the
moving party is entitled to judgment as a matter of law. Maiden v Rozwood, 461 Mich 109, 120;
597 NW2d 817 (1999).
On appeal, plaintiff only challenges the dismissal of her retaliation claim. To establish a
prima facie case of retaliation under the Civil Rights Act, MCL 37.2701(a), a plaintiff must
(1) that the plaintiff engaged in a protected activity, (2) that this was known by the
defendant, (3) that the defendant took an employment action adverse to the
plaintiff, and (4) that there was a causal connection between the protected activity
and the adverse employment action. To establish causation, the plaintiff must
show that his participation in activity protected by the CRA was a ?significant
factor? in the employer?s adverse employment action, not just that there was a
causal link between the two. [Barrett v Kirtland Community College, 245 Mich
App 306, 315; 628 NW2d 63 (2001) (internal citations and quotations omitted).]
Thus, a plaintiff cannot prevail on a retaliation claim under the CRA without establishing that
she engaged in an activity protected under the act. Under MCL 37.2701(a), a protected activity
is opposition to a violation of the act, or participation in an investigation, proceeding, or hearing
under the act.
Here, plaintiff alleged that she opposed a violation of the act, i.e., age discrimination, and
that his was evidenced by her January 29, 2002 letter indicating that she could not work in a
hostile environment where she was denied promotions and placed under a new supervisor.
Plaintiff presented no evidence that she complained about age discrimination or failure to follow
company policies prior to her dismissal. Where a plaintiff does not show that adverse treatment
constituted a violation of the CRA, the plaintiff cannot establish that the employer retaliated
against him or her for opposition to a violation of the CRA. Barrett, supra at 323-324. The trial
court properly granted defendant?s motion for summary disposition where plaintiff failed to
present evidence that she engaged in a protected activity.
/s/ Christopher M. Murray
/s/ David H. Sawyer
/s/ Michael R. Smolenski


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Beavan, Noesi to start season in rotation
Beavan, Noesi to start season in rotation
Those two will both be breaking camp as Major League starters for the first time, along with veterans Felix Hernandez, Jason Vargas and Kevin Millwood.
Beavan, 23, started 15 games after a midseason promotion last year for Seattle. Noesi, 25, made two spot starts last year for the Yankees, but was primarily a reliever in his rookie season.
"It was good to hear that," Noesi said after learning he'd won a spot. "I've been waiting for that since whenever I started in baseball, to start in the big leagues."
Although he was with the Yankees last season, Noesi is eager to take a larger role in Seattle.
"It's big," he said. "I have just one year of playing in the big leagues. I'd like to be part of the team. I just have to keep me in there now."
Noesi will pitch the first exhibition game in Japan against the Hanshin Tigers on Sunday, while Beavan will be held in reserve in case something should happen to Felix Hernandez or Jason Vargas prior to the March 28-29 regular-season openers against the A's in the Tokyo Dome.
Beavan said it was a relief to win a job after all the competition that emerged for the rotation with the trade for Noesi and signings of Millwood and Hisashi Iwakuma.
"It's pretty hard for a rookie to look around and see everybody they signed and still think you have a shot," he said, "but being as competitive as I am, I never let anything try to get to me too much and I tried to stick to what I do and just come to camp ready to throw strikes and keep the ball down."
He succeeded with both and now has a starting role, though he and Noesi won't likely pitch in a regular-season game until the April 9-12 road trip to Texas.


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Dish has secured a content licensing deal with A&E Networks for the company's yet-to-be announced web TV streaming service, reports Re/code.
The service was first rumored way back in 2012 as part of an overall plan by Dish to become more competitive than rival DirecTV as well as cable television providers like Comcast. The biggest advantage of an IP-based TV service would be that you can access it regardless of your location, something that you can't really do with competing services. (The one exception would have been Aereo, but that service shut down after last month's Supreme Court decision.)
The licensing deal will apparently bring all currently airing TV shows from A&E's stable of cable stations, including A&E, History, H2, Lifetime, fyi, and the Military History channel. But customers won't be able to access a direct linear feed from those channels. Instead, they will be given on-demand access to shows.
Dish also previously signed a similar deal with Disney to air content from Disney-owned cable channels via its unlaunched web TV service. However, if Dish really wants to make a go at a competitive IP-based TV service, it'll need to sign on plenty of other media partners — namely Viacom, which owns Comedy Central, Spike, and MTV.


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Open Source Malaria (OSM, pronounced "awesome") is aimed at finding new medicines for malaria using strictly open source principles, embodied in the 6 Laws of Open Research.
This wiki describes where the project is up to, and summaries of relevant data. For other aspects of the project, see the main Project Landing Page.
If you would like to improve these wiki pages, you will need to get an OpenWetWare account (which is easy).
About the Malaria Project
A human-friendly summary of the science to date can be found in The Story So Far.
A description of what open research is can be found in the Open Research Page
Aggregated data on various branches of the project so far can be found on the Compound Series Page.
What's Happening and What's Needed
Daily updates (including the To Do list) can be accessed through the project's main hub.
At the moment (July 2013) there is most activity on Series 2, with the raw data in this lab notebook.
The main resource needed in the OSM project is experimental input, particularly chemical compound synthesis. Anyone can make compounds, get them tested and contribute data. You don't have to be a research scientist - maybe you're an undergrad doing a lab.
If you have no access to a lab, there are a number of important admin things needed, labelled as such on the To Do List.
Open Source Drug Discovery More Generally
A one-day meeting on open source drug discovery for malaria was held in February 2012. General issues surrounding the feasibility of open source drug discovery were discussed, followed by more specific malaria-related ideas. These talks are gradually going up on YouTube with annotations, and they frame many of the relevant issues, for example the landscape of drug discovery in neglected diseases, and whether patents are necessary in drug discovery. An important message is that open source drug discovery is where anyone may participate in driving the research, which is different from a more general use of the word "open" where data are made freely available, but perhaps after a delay which essentially prevents participation by others.
How We Run the Project
The way the project is run is one of the novelties, though as with everything in this project nothing is static and advice is always welcome on improvements. Raw experimental data are recorded in an online, openly-readable electronic lab notebook. The Synaptic Leap is being used to discuss ideas and results, as well as plan future work. The project's Google+ page is a light way to keep up with developments and discuss. The project's Twitter feed is a broadcast mechanism for updates. LinkedIn as used in the past on another project as a way of connecting with relevant experts, but has not been used much so far in this project. A wiki (that includes this page) is used to host the current overall project status. Updates on the project's progress can also be found at our Facebook page, and this also a place for interaction. If you wish to participate in this project, you can sign up to all these sites, and you would then be sent the Twitter/G+ passwords so you can used the same accounts.
Why Take Part?
What of motivations? Why would people want to contribute to this project? Partly to solve a problem. Partly to be involved with quality science that is open, and hence subject to the most brutal form of ongoing peer-review. Partly for academic credentials since regular peer-reviewed papers will come from the project. Partly to demonstrate competence publicly - open science is meritocratic and status-blind. Perhaps a mixture of all these things.
A competition is possible in the future, i.e. with a cash prize. Progress towards a very promising lead compound series has been rapid, but there is a long road to a compound that looks sufficiently promising that it moves towards clinical trials. There's a lot of tweaking needed, and perhaps even the move to another series. It is not obvious what will happen. It is certain the project will need a lot more input than it has received to date. A prize may increase traffic and input. The competition would be teamless, however, awarded based on performance of individuals within a group where everything is shared. Not sharing data or ideas leads to disqualification. Such a competition is difficult to judge, difficult to award, and hence almost certainly worth doing. More about this is here.
A final point - the project is open. Nobody owns it. Those people most active in the project lead it while they are active. If you wish to contribute, in any capacity, please do so. There is no need to "clear" anything with existing project members by email first. To date is has been very common for current participants to receive questions/suggestions from people by email, which is to be discouraged. In the development of Linux, the need for Linus Torvalds to approve everything caused a serious bottleneck, and the observation that "Linus doesn't scale". Nobody scales, but the team does. So it's more efficient if all the project discussions are held publicly. Many people do not like this idea. In science the idea of "beta testing" something is alien. When data are released in science there is an expectation that the data are correct, and essentially finished. This project eschews this view. All data are released immediately, all discussions are public, anyone can participate.
This is a wiki, meaning it's meant as a site for how projects are going. For discussion try The Synaptic Leap, or the project's Google+ page, and for updates try the Twitter feed and Facebook Page. If you would like access to the Twitter or G+ accounts because you've become a member of the project please email firstname.lastname@example.com.
Relevant papers are available in the Mendeley group, to which anyone can add.
The OSM project's licence unless otherwise stated is CC-BY-3.0 meaning you can use whatever you want for whatever purpose, provided you cite the project.


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I relaunched our partner site, Minnesota KidVid this past week. The Minnesota KidVid website was the idea of Julian West, building upon his original Axel's Treehouse website and expanding to include other Minnesota TV programs from the era of live programming. Julian did the initial leg work several years ago for the new site, but […]
In the Casey Jones Shop
Visit our Lunch With Casey Shop
for DVDS and more


text:
Injury update: R.J. Mattes is questionable with an ankle injury.— New England Patriots (@Patriots) August 17, 2013
ESPNBoston.com's Mike Reiss tweeted "Patriots injury update: OT R.J. Mattes (ankle), return questionable". Additionally, Boston Hearald's Jeff Howe tweeted "R.J. Mattes is questionable to return after an ankle injury".
The Offical Twitter Account of the New England Patriots tweeted "Injury update: R.J. Mattes is questionable with an ankle injury". Pro Football Focus's Steve Palazzolo also tweeted: R.J. Mattes led the Patriots offense with 56 snaps and graded at +1.8 last week.


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Global Synthetic Carbon Film Potentiator Industry 2022 Market Research Report analyzed the current state in the definitions, classifications, applications and industry chain structure. The report also focuses on the development trends as well as history, competitive landscape analysis, and key regions etc in the international markets.
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Elite's Globalist Manifesto of Rules
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Financial Tyranny: 'We the People' Are the New Permanent Underclass in America
David Icke: All The World Is Mad (Video)
David Wilcock: The Downfall Of The Cabal & Something Big Is Coming (Video)
Liberals are Completely Unravelling and Admitting Insanity!
Robert David Steele Deep State dissolved with Jason Liosatos
Tucker Carlson Points Out Exactly Why it's the 'End of the Clinton Era, Finally and Completely'
Greg Hunter, "Fed Fears New Record High Credit Bubble"
It Has Now Been Confirmed: It Was Not Russia...it Was Obama!
Elite's Globalist Manifesto of Rules
Financial Tyranny: 'We the People' Are the New Permanent Underclass in America
Major Gas Line Explodes in Mich; Sabotage Suspected – North Korea Sleeper Cells?
Is the Messiah Coming This Year? Iran-Saudi War Fulfills Ancient Prophecy!
Truth story behind Donald Trump, Asia Trip & Zimbabwe
'New York' Looks Like a Skid Row Bum - Indigenous People Never Gave Away Their Land
Expose the Network and You will find the Missing People
Venezuela Arrests President of U.S.-Based Refiner Citgo
I smiled at Putin's expression…. How about some newsy coffee this morning?
How Modern Empire Uses "Awards" to Keep Servants Loyal
A Time For Thanksgiving, A Time For Preparation
Alarming! Pentagon Insider on "The Deeper State"
A Swarm Of Earthquakes Beneath The San Andreas Fault Is Making Scientists Nervous
A Culture That Trained Men Not to Respect Women but to Respect Feminism
The Mystery of the Origin of Gold Deposits Gets Very Deep Answer
"This Is Who's Behind Slaughterbots Viral Video Campaign" {Warning: Do Not Sign the Petition} (Video)
"A New Theory of History – Moronism: A History of Idiocy"
You Won't Believe What 900 Scientists Just Concluded About Major Liberal 'Issue', Dems in Panic Mode
It's Happening! President Trump Is About to Smash Thousands of Welfare Leeches, Obama Is Furious
How Crazy Have Liberals Gone? Here's What They Say About the US Flag


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Lebanon: Information on property confiscations by the various militias and the effort to return them to their owners
1 May 1993 | Publisher: Canada: Immigration and Refugee Board of Canada | Document type: Query Responses


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After more than a decade without a general state tax increase, most of the Democrats running for governor say they would consider raising taxes to help solve Minnesota's financial problems.
Candidates traditionally go to great lengths to avoid the "T" word, but several DFL gubernatorial hopefuls say the looming budget shortfall requires a different approach.
When DFL House Speaker Margaret Anderson Kelliher kicked off her gubernatorial campaign last week near Mankato, state Republican Party officials sent out a news release warning that the Minneapolis Democrat is out to raise taxes.
It wasn't exactly a shocking revelation. All of the DFL candidates for governor have been talking about tax increases. Kelliher just isn't ready to offer many specifics.
"I think it's difficult to say exactly what it is going to be," Kelliher said. "But there is no one in this race who can tell Minnesotans that they'll be able to balance the budget without severely harming the valuable things that Minnesotans care about -- education, health care, the state-local relationship -- if there is not some amount of revenue raised at the state level."
Kelliher might not know the amount of revenue, but she has at least one source in mind. She said Minnesotans who are earning over $500,000 a year need to pay their fair share of income tax. That's the income group former U.S. Sen. Mark Dayton said he would also target.
"The wealthiest 10 percent of Minnesotans pay only about two-thirds of their proportionate share of their incomes in state and local taxes as the next 80 percent," Dayton said. "That is fundamentally wrong."
Minnesota's next governor won't take office until January 2011, but some early predictions suggest the new chief executive will face a state budget deficit of between $4.5 billion and $7 billion -- anywhere from 14 to 22 percent of the budget.
"I think there are a lot of people in this state that make some pretty good money that wouldn't mind at all paying a little more in tax."State Rep. Tom Rukavina
State Rep. Tom Rukavina of Virginia said the need to raise new revenue won't be the question. He said the only question is: who will pay?
"People make money even during a recession in the stock market, and I think Minnesota has a reputation of helping their neighbors, of being compassionate," Rukavina said. "I think there are a lot of people in this state that make some pretty good money that wouldn't mind at all paying a little more in tax."
Rukavina wants to reverse income tax cuts enacted in 1999 and other DFL candidates agree. State Rep. Paul Thissen of Minneapolis said tax increases alone won't close the entire deficit, but he said new revenue is essential. Thissen and others say expanding the sales tax to clothing or personal services that are currently untaxed should also be discussed.
"I think that we need to sit down and talk about everything," Thissen said. "I don't want to necessarily point to a particular preference of mine right now. I think we need to put everything on the table."
DFL candidates say they're just trying to be honest about taxes, but history shows that politicians can pay a big price for such honesty. Minnesota's Walter Mondale took a hit in the 1984 presidential race when he went with the open approach.
"Mr. Reagan will raise your taxes, and so will I. He won't tell you. I just did," Mondale said during his acceptance speech at the Democratic National Convention in 1984.
Dan Hofrenning, a political science professor at St. Olaf College in Northfield, said Mondale's remark helped Republicans link Democrats with high taxes and big government and it was a winning formula for decades. Hofrenning said it's unusual to hear Democrats again talking openly about tax increases, but he said the political landscape is changing.
"I think Minnesotans and American citizens, they haven't become thoroughly enamored with big government and significant tax increase, but they're more open to it," Hofrenning said. "If they can be convinced that government can bring a more prosperous and just society, I think there's a willingness to be open to some tax increases."
State Sen. John Marty of Roseville has taken political heat before for his stand on taxes. In 1999, he was the lone vote against legislation to lower income taxes. Marty said much of today's budget trouble began with that bill. He said he was honest with voters then and he's being honest with them now.
"I think most voters would rather have somebody who can talk to them as adults and say here's the choices we have, instead of saying, oh, I'm not going to use the tax word because it could cost me some votes," Marty said.
DFL gubernatorial candidates will detail their views on taxes and other issues during a forum Thursday night in St. Cloud. There are currently 10 Democrats in the race, and more are expected to join.