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Despite representing perhaps the simplest form of memory, habituation is not yet well understood mechanistically. We used a reduced preparation to analyze the neurobiological mechanisms of persistent habituation of a simple behavior, the defensive withdrawal reflex of the marine snail Aplysia californica. This preparation permits direct infusion of drugs into the abdominal ganglion during training via a cannula in the abdominal artery. Using siphon-elicited gill withdrawal, we demonstrate habituation of withdrawal that persists for 1-6 hr after repeated, spaced blocks of habituating stimulation. This form of habituation exhibits site specificity and requires protein synthesis because it is blocked by the presence of anisomycin, a protein synthesis inhibitor. We also find that habituation of gill withdrawal requires protein phosphatase activity, because it is blocked by okadaic acid, an inhibitor of protein phosphatase. Finally, habituation of gill withdrawal requires activation of NMDA-type and AMPA-type postsynaptic receptors within the abdominal ganglion, because it is blocked by infusion of dl-2-amino-5-phosphonovaleric acid or 6,7-dinitroquinoxaline-2,3-dione. The requirement for activation of postsynaptic glutamatergic receptors indicates that homosynaptic depression, an exclusively presynaptic mechanism that has been implicated previously in habituation in Aplysia, does not play a significant role in persistent habituation of the withdrawal reflex. Our results indicate that postsynaptic mechanisms, possibly including modulation of glutamate receptor function, play a major, heretofore unsuspected, role in habituation in Aplysia.
- Aplysia
- habituation
- long-term memory
- protein synthesis
- protein phosphatase
- okadaic acid
- glutamate receptors
- postsynaptic processes
Habituation, perhaps the simplest form of learning, is the decrement in an animal's response to repeated presentations of a (typically innocuous) stimulus, in which the decrement cannot be ascribed simply to injury, receptor adaptation, or motor fatigue (Thompson and Spencer, 1966). Habituation is a nearly universal form of learning, having been observed in animals ranging from roundworms (Rankin et al., 1990) to humans (Geer, 1966). Despite its apparent simplicity and universality, our understanding of the neurobiological basis of habituation remains incomplete. A generation ago, it was widely agreed that habituation, at least for certain simple reflexes in invertebrate organisms, was attributable to a presynaptic mechanism: homosynaptic depression of transmitter release from the terminals of the sensory neurons that mediate the reflexes (Zucker, 1972; Castellucci and Kandel, 1974; Krasne, 1976; Kandel, 1978). However, more recent work has indicated that inputs from extrinsic modulatory pathways are critical to the full expression of behavioral habituation, even in invertebrates (Krasne and Glanzman, 1995; Krasne and Teshiba, 1995; Stopfer and Carew, 1996).
Habituation is commonly observed to have both short-term and long-term forms. Cellular correlates of long-term habituation (LTH) of the defensive withdrawal reflex of Aplysia (Carew et al., 1972; Carew and Kandel, 1973) have been identified. Behavioral training that results in habituation persisting for ≥24 hr causes a long-term depression of transmission as sensorimotor synapses (Castellucci et al., 1978), as well as a significant reduction in active zones and varicosities associated with sensory neurons in the abdominal ganglia of habituated animals (Bailey and Chen, 1983, 1988).
Although the cellular expression of LTH of the withdrawal reflex in Aplysia is attributable, at least in part, to long-term presynaptic changes, the locus of the induction of the cellular changes is uncertain. Are the long-term changes in sensory neuron transmitter release and structure that are associated with LTH induced, as well as expressed, presynaptically? Or, rather, are the long-term cellular changes induced by postsynaptic processes? Here, we show that postsynaptic mechanisms, particularly activation of postsynaptic glutamate receptors, play a critical role in inducing long-lasting habituation of the withdrawal reflex in Aplysia.
The habituation we report here persists for 1-6 hr after the last block of stimuli and depends on protein synthesis (see Results). A requirement for protein synthesis has been typically regarded as an essential characteristic of long-term memory (Davis and Squire, 1984; Goelet et al., 1986). However, we decided not to refer to the present form of habituation as long term. Previous studies of LTH in Aplysia have reported habituation that persisted for 24 hr to weeks (Carew et al., 1972; Carew and Kandel, 1973; Castellucci et al., 1978; Stopfer et al., 1996), and it is possible that LTH of the duration achieved in these previous studies will ultimately prove distinct, mechanistically, from the habituation we demonstrate here. We will therefore refer to the prolonged habituation shown in the present experiments as "long-lasting habituation (LLH)."
Parts of this work have been published previously in abstract form (Ezzeddine and Glanzman, 2001, 2002).
Materials and Methods
Animals. Adult Aplysia californica (75-150 gm) were obtained from a local supplier (Alacrity Marine, Redondo Beach, CA). Animals were housed in a 50 gallon aquarium filled with cooled (14°C), aerated artificial seawater (ASW) (Instant Ocean; Aquarium Systems, Mentor, OH). All animals were housed for at least 24 hr before the start of the experiment.
Preparation. An animal was initially anesthetized with an injection of 60-80 cc of isotonic MgCl2 into the animal's foot. The animal was then placed, ventral side up, in a dissection tray. A longitudinal incision was made along the length of the foot. The body wall was pinned back to expose the digestive organs and nervous system. The digestive system was removed to gain access to the nervous system. The CNS was drawn toward the tail, and most of the peripheral nerves were cut. The head of the animal, together with the buccal ganglia, was then removed. The rest of the animal, including the mantle shelf, gill, and siphon, together with the tail and the entire CNS (minus the buccal ganglia), was preserved (Fig. 1). The CNS was left connected to the siphon and gill via the siphon and branchial nerves of the abdominal ganglion. The animal was transferred to another dissection tray filled with 50% MgCl2-50% normal ASW. The artery leading to the abdominal ganglion was cannulated with polyethylene tubing (0.024 outer diameter, 0.011 inner diameter; Intramedic, Parsippany, NJ), which was connected to a peristaltic pump (P720; Instech, Plymouth Meeting, PA). During experiments, the abdominal artery was perfused with aerated ASW (15°C) via the cannula at a rate of ∼1 ml/hr until the start of the experiment (onset of the pretests), at which point the perfusion rate was decreased to 0.5 ml/hr. This perfusion rate was then maintained throughout the experiment. The cannula in the abdominal artery was also used to selectively administer drugs to the abdominal ganglion (below). After cannulation of the abdominal artery, the preparation was transferred to a Lucite experimental chamber filled with normal ASW and pinned, dorsal side up, to the Sylgard-lined bottom of the chamber. The siphon was not pinned but was left to move freely. The afferent vein of the gill was cannulated with polyethylene tubing (above) and perfused with chilled, aerated ASW via a peristaltic pump (Dynamax RP1; Rainin, Oakland, CA) at a rate of 100 ml/hr. The cannula in the afferent vein was secured with a surgical silk suture, and a force transducer (model 1040 or 1030, ADInstruments, Grand Junction, CO) was connected to the suture with a small metal hook. The force produced by movement of the transducer was calibrated by hanging gram weights from the metal hook. Habituating stimuli (see below) were delivered to the siphon via pairs of Teflon-insulated platinum wires (0.005 mm in diameter; catalog #773000; A-M Systems, Carlsborg, WA). The uninsulated tips of the wires were inserted into the siphon at its base. In the within-preparation experiments (see below), a platinum wire was inserted into each side of the siphon, and a ground wire was placed in the bath. The rest of the preparation was perfused with normal ASW (13-15°) at a rate of 1 l/hr throughout the experiment.
After the intensity of the siphon stimuli to be used for an experiment had been determined (see below), the preparation was rested for 2 hr, during which time the afferent vein of the gill and the rest of the preparation were perfused with normal ASW to completely wash out the MgCl2. This perfusion was maintained for the rest of the experiment. At the end of the 2 hr rest period, a single stimulus (at the predetermined intensity for habituation) was delivered to the siphon. Preparations that did not respond to this initial stimulus with a visible gill contraction were discarded. Furthermore, data from preparations from the experiments in which a within-preparation design was used (below) were only accepted if the posttest response of the "untrained" site was at least 80% of the pretest response (see Experimental design).
Shipments of Aplysia from our local supplier varied significantly according to whether or not preparations from the animals expressed long-lasting habituation. In some cases, these differences were seasonal and may represent differences in the behavioral state of the animals. In other cases, the differences may have been related to the health of the animals. Because of the variability in the amount of habituation exhibited by preparations from different shipments of animals, we were obliged to institute a control criterion to ensure that animals in a given shipment were capable of expressing LLH. On the basis of the results of pilot experiments, we decided that preparations could be considered to have undergone long-lasting habituation if the posttest response (at either 6 hr or 60 min after the end of training; see below) of the "trained" site to the habituating stimulus was ≤70% of the pretest response. Using this criterion, we ran an initial control experiment with one animal from each shipment. If that control preparation did not habituate by ≥30%, the entire shipment of animals was rejected.
Siphon stimulation. Both habituating and test stimuli were brief trains (500 msec, 25 Hz) of current pulses (10 msec duration) generated by an electronic stimulator (S48/S88; Grass Instruments, Quincy, MA); the stimuli were delivered to the siphon via implanted platinum wires (see above). The intensity of the stimuli was calibrated for each preparation to be just suprathreshold for reliably eliciting gill withdrawal. The intensity of the stimuli applied to the untrained site in experiments using the within-preparation design (below) was determined in the same way and was also fixed for a given experiment.
Drugs. All drugs were purchased from Sigma-Aldrich (St. Louis, MO). The drugs were dissolved in either normal ASW or DMSO as stated in Results. The drugs were infused directly into the abdominal ganglion via a cannula in the abdominal artery (above). The anisomycin was infused into the abdominal ganglion for 45 min to 1 hr before the onset of the pretests; okadaic acid (OA) and 1-nor-okadone were infused for 2 hr before the onset of the pretests. Infusion of anisomycin, okadaic acid, and 1-nor-okadone was maintained throughout those experiments in which they were used. We performed two sets of dl-2-amino-5-phosphonovaleric acid (APV) experiments. In the first set of experiments, APV was infused into the abdominal ganglion for 45 min to 1 hr before the onset of the pretests and was present throughout the experiments, as in the experiments involving anisomycin, okadaic acid, and 1-nor-okadone. In the second set of experiments, APV was present only during training; both the pretests and posttests were performed in ASW. Our results for these two sets of APV experiments were very similar (see Results). (Notice that both okadaic acid and 1-nor-okadone took a prolonged period of time for wash in, so it was not feasible to have these compounds present only during training. We did not perform experiments with the protein synthesis inhibitor present only during training.) 6,7-Dinitroquinoxaline-2,3-dione (DNQX) was infused at a rate of 1 ml/hr starting immediately after the pretests; it was then washed out of the abdominal ganglion with normal ASW after the end of habituation training.
Experimental design. There were two basic experimental designs. In our initial set of experiments (between-preparation design), there were two separate groups of preparations: an experimental group that received habituation training and a control group that received only test stimuli. In subsequent experiments, we used a modified design in which each preparation served as its own control (within-preparation design). In the experiments using the within-preparation design, two different sites on the siphon were stimulated: one of the sites (trained) received habituation training, whereas the other site (untrained) received only test stimuli (Fig. 1). In the between-preparation experiments, each preparation initially received three pretests at a rate of one per 10 min. Fifteen minutes after the third pretest, some preparations (trained) received habituation training. The training protocol consisted of four blocks of 20 stimuli [interstimulus interval (ISI) of 30 sec]. There was a 90 min rest period between each block of habituating stimuli. After the fourth training block, there was a 6 hr rest period, and then the preparation received three posttests at a rate of one per 10 min. Control (untrained) preparations received three posttests at the same rate 12 hr after the pretests. In the within-preparation experiments, the side of the siphon that was chosen to be the trained site was alternated systematically between left and right sides. The trained site initially received three pretests at a rate of one per 10 min. Ten minutes after the third pretest to the trained site, the other side of the siphon (untrained site) received a single pretest stimulus. Fifteen minutes after the pretest stimulus to the untrained site (25 min after the third pretest to the trained site), the trained site received habituation training. The training protocol was identical to that used for the between-preparation experiments. In the within-preparation experiments, there was 60 min rest period between the final block of training stimuli and the onset of the posttests. Then, the trained site received three posttests at one per 10 min. Ten minutes later, the untrained site received a single posttest.
A somewhat different within-preparation experimental protocol was used in one set of the experiments with the drug APV and in all of those with the drug DNQX (see Results). The altered protocol was used to ensure that the drugs were only present during training and were completely washed out after training. In these experiments, the trained site received two pretests at one per 5 min; 5 min later, the untrained site received two pretests at one per 5 min. Then, there was a 1 hr pretraining rest period during which the drug (APV or DNQX) was infused into the abdominal ganglion via the abdominal artery, after which habituation training of the trained site commenced. After the fourth (final) block of habituating stimuli was presented, there was a 2 hr posttraining rest period to permit thorough washout of the APV-DNQX.
Data acquisition and evaluation. The force transducer used for measuring gill contractions (above) was connected through a bridge amplifier (ML110; ADInstruments) to a digital data acquisition system (MacLab 4s/2e; ADInstruments). The peak amplitude (maximum - minimum) of each gill contraction was determined with the data acquisition system. The minimum was the baseline response recorded when the gill was in a relaxed state 1 sec before stimulation. The maximum was the peak amplitude of the response within 20 sec after the siphon stimulus. If the amplitude of the response declined by ≥80% from the initial peak within 20 sec after the siphon stimulus, the response was considered complete; subsequent responses within the 20 sec period were not counted. Notice that, during training, the 30 sec ISI was not always sufficient to permit the gill to fully relax after a response. In these cases, the minimum value for a particular response was the level of gill contraction before the onset of the training block.
Statistics. The pretest gill response in the experiments using the between-preparation design was the mean value for the three pretests. This value was set to be equal to 100%, and the rest of the gill responses were normalized to the pretest value. There were also three posttests, and the mean of the three tests was used for the statistical comparisons. In the experiments using the within-preparation design, the pretest gill response for the trained site was also the mean value for the three pretests. The trained site also received three posttests. There were only a single pretest and a single posttest, however, for the untrained site in the within-preparation experiments. The pretest value (mean of three pretests for the trained site or the single pretest value for the untrained site) was set to be equal to 100%. The subsequent values for the gill responses to stimulation of the trained-untrained sites were normalized to 100%. All responses are expressed as the normalized mean response ± SEM. Nonparametric statistical tests were used for most comparisons. Wilcoxon tests were used for paired comparisons and Mann-Whitney tests were used for unpaired comparisons, unless otherwise indicated. All significance values reported represent two-tailed levels of significance.
Long-lasting habituation of gill withdrawal in a reduced preparation
Our initial set of experiments used a between-preparation design (see Materials and Methods). There were two groups. One group (trained; n = 9) received four blocks of habituating stimulation spaced at 90 min intervals. Each training block comprised 20 bouts of weak electrical stimulation of the siphon with a 30 sec ISI. Another group (control; n = 13) received only the pretest and posttest stimuli. When tested 6 hr after the final (fourth) training block, gill withdrawal in the trained preparations exhibited significant habituation (mean posttest response, 34.6 ± 5.9%; p < 0.004 for the comparison with the pretest value, which equaled 100% in this and all other experiments; see Materials and Methods) (Fig. 2). The magnitude of gill withdrawal did not change in the control preparations (mean posttest response, 91.5 ± 12.3%; p > 0.05 for the comparison with the pretest response; data not shown). Furthermore, the difference between the posttest responses in the trained and control preparations was significant (p < 0.002). The lack of a significant difference in the magnitude of the gill reflex in control preparations during the experiments indicates that the decrement in the gill reflex in the trained group could not be attributable to deterioration in the health of the preparation over the >12 hr of the experiment. This suggests, in turn, that the training produced LLH in the trained group. It still might be argued, however, that the response decrement in the trained group was attributable to motor or muscle fatigue. We therefore modified our experimental protocol to exclude motor-muscle fatigue as a possible explanation for the response decrement exhibited by the trained preparations.
Reduced preparation exhibits site-specific long-lasting habituation of gill withdrawal
In the rest of our experiments, we used a within-preparation design. Two stimulation sites on the siphon were used rather than one. One site (trained) received the spaced blocks of habituating stimuli, whereas the other site (untrained) received only the pretest and posttest stimuli. The testing and training protocol were similar to those in the between-preparation experiments (above), with the following exceptions. The untrained site received only a single pretest and a single posttest. Furthermore, the rest period between the end of the habituation training and the start of the posttest stimulation of the trained site was shortened from 6 to 1 hr. This was done to streamline the experiments. One hour after the end of training, the response of the gill to stimulation of the trained site was significantly reduced compared with the pretest response of the gill (mean posttest response, 64.8 ± 4.2%; p < 0.003 for the comparison with the pretest response) (Fig. 3A). In contrast, the posttest response of gill to stimulation of the untrained site was significantly greater than the pretest response (mean posttest response, 115.1 ± 7.3%; p < 0.04 for the comparison with the pretest response). This result suggests that the habituation training of the trained site induced modest sensitization of the untrained site within the same preparations. The response of the gill to stimulation of the trained site on the first of the three posttests (77.4 ± 7.6%) was also significantly less than the posttest response to stimulation of the untrained site (p < 0.02; n = 10). (Notice that the interval between the pretests and posttests is identical for this comparison.) The difference between the posttest responses to the trained and untrained sites in these experiments cannot be ascribed to motor or muscle fatigue, because the same preparation was used to control for the effects of training on the neuromuscular components of the reflex. These results therefore demonstrate site-specific habituation of siphon-elicited gill withdrawal in a reduced preparation.
The amount of habituation was greater in the between-preparation experiments than in the within-preparation experiments (p < 0.002 for the comparison between the posttest value for the between-preparation experiments shown in Fig. 2 and the posttest value for trained-site stimulation in Fig. 3A). This difference may indicate that the longer interval between the termination of training and the posttests in the between-preparation experiments resulted in greater consolidation of LLH. However, because these two sets of experiments were performed at different times using animals from different shipments, it is also possible that the greater apparent habituation in the between-preparation experiments was attributable to a seasonal or animal shipment effect.
Long-lasting habituation of gill withdrawal requires protein synthesis
A hallmark of long-term memory across a range of preparations and different forms of learning is that it depends on the synthesis of new proteins (Davis and Squire, 1984; Goelet et al., 1986). To test whether LLH of gill withdrawal in our reduced preparation also exhibits a requirement for protein synthesis, we infused the protein synthesis inhibitor anisomycin (30 μm in ASW) into the abdominal ganglion before the start of an experiment. This concentration of anisomycin has been shown previously to inhibit protein synthesis by >95% in the abdominal ganglion of Aplysia (Schwartz et al., 1971) and in sensory and motor neurons from the abdominal ganglion (Montarolo et al., 1986). The protein synthesis inhibitor was present throughout the experiment. Anisomycin blocked LLH of the withdrawal reflex to stimulation of the trained site. After habituation training in anisomycin, the response of the gill to stimulation of the trained siphon site was enhanced (posttest response, 126.7 ± 14.6%; p > 0.09 for the comparison with the pretest response) (Fig. 3B), although this enhancement was not significant. The posttest response of the gill to stimulation of the untrained site (121.0 ± 15.3%) was also enhanced, although again not significantly (p > 0.1). Furthermore, the difference between the first posttest response to stimulation of the trained site (160.5 ± 18.7%) and the posttest response to stimulation of the untrained site was not significant (p > 0.09; n = 9). A crucial comparison is that between the posttest values for trained site stimulation in the experiments of Figure 3, A and B. This is because one might argue that the apparent blockade of LLH by anisomycin actually results from the fluctuations in the capacity for LLH that we observed among shipments of animals (see Materials and Methods). However, the two sets of experiments in Figure 3, A and B, were performed at the same time using animals from the same shipments. Therefore, the experiment in Figure 3A represents a control for the capability of the preparations used in the experiment in Figure 3B to undergo habituation. The difference between the posttest response to trained-site stimulation in the experiments shown in Figure 3, A and B, was highly significant (p < 0.001). This result supports the notion that LLH was blocked in the experiments of Figure 3B by the presence of the protein synthesis inhibitor.
Long-lasting habituation of gill withdrawal requires protein phosphatase activity
A potential cellular mechanism for LLH of gill withdrawal is long-term depression (LTD) of the monosynaptic connection between sensory and motor neurons in the abdominal ganglion (Lin and Glanzman, 1996). LTD of synapses in the mammalian hippocampus and cortex is attributable, in part, to activation of protein phosphatases (Mulkey et al., 1993; Bear and Abraham, 1996). To test for a possible role of protein phosphatase activity in habituation of gill withdrawal, we infused okadaic acid, a selective inhibitor of protein phosphatases 1 and 2A (PP1 and PP2A) (Cohen et al., 1990), into the abdominal ganglion. OA (1 μm in ASW with 0.1% DMSO) was infused into the abdominal ganglion beginning 2 hr before the first pretest and was present throughout the experiment. The presence of OA blocked LLH (Fig. 4A). The posttest response to stimulation of the trained site (108 ± 13.9%) was not significantly different from the pretest trained-site response (p = 1.0). Furthermore, there was no significant difference between the first posttest response to stimulation of the trained site (118.4 ± 12.1%) and the posttest response to stimulation of the untrained site in OA (133.3 ± 23.9%; p > 0.8; n = 13). The OA did not appear to affect the response of the gill to stimulation of the untrained site over the course of the experiment. The posttest response to untrained-site stimulation was not significantly different from the pretest response (p > 0.5). To control for nonspecific actions of OA, we tested the effect of the control compound 1-nor-okadone (1 μm in ASW with 0.1% DMSO), an analog of OA that does not inhibit phosphatases (Mulkey et al., 1993), on LLH of gill withdrawal (Fig. 4B). The experiments using 1-nor-okadone were performed at the same time as the experiments using OA and used animals from the same shipments. Infusion of 1-nor-okadone into the abdominal ganglion did not inhibit LLH. The posttest gill response to stimulation of the trained (68.5 ± 4.7%) site was significantly lower than the pretest response (p < 0.008; n = 8). The 1-nor-okadone treatment did not appear to affect the response of the gill to stimulation of the untrained site. The posttest response to stimulation of the untrained site (117.6 ± 22.1%) was not significantly different from the pretest response (p > 0.8). The posttest response of the gill to trained-site stimulation was significantly less in the 1-nor-okadone experiments than in the OA experiments (p < 0.03), a result that further supports the idea that protein phosphatase activity is essential for LLH.
Long-lasting habituation of gill withdrawal requires activation of NMDA-type receptors
The CNS of Aplysia possesses postsynaptic NMDA-type receptors (Dale and Kandel, 1993; Lin and Glanzman, 1994; Conrad et al., 1999). Activation of postsynaptic NMDA-type receptors has been shown previously to be necessary for classical conditioning of the withdrawal reflex in Aplysia (Murphy and Glanzman, 1997, 1999; Antonov et al., 2003). We wanted to test whether activation of NMDA-type receptors was also required for LLH of the withdrawal reflex. For this test, we infused the specific NMDA receptor-antagonist APV (150 μm in ASW) (Collingridge et al., 1983; Lin and Glanzman, 1994; Conrad et al., 1999) into the abdominal ganglion. The drug was introduced into the abdominal artery 1 hr before the first pretest and was present throughout the experiment. The presence of APV blocked LLH to the habituating stimuli (Fig. 5A). There was no significant difference between the pretest and posttest responses to stimulation of the trained site after training in APV (posttest response, 110.1 ± 15.4% of the pretest response; p > 0.5; n = 10). Furthermore, there was no significant difference between the first posttest response to trained-site stimulation (138 ± 16%) and the posttest response to untrained-site stimulation (123.6 ± 15.1%; p > 0.5). APV treatment did not appear to affect gill withdrawal to stimulation of the untrained site, because it did not change significantly during the experiment (p > 0.2 for the comparison between the pretest and posttest responses to untrained-site stimulation). We performed control experiments using animals from the same shipments that were used in the APV experiments. In the control experiments, the abdominal ganglion was continually infused with normal ASW. We observed normal LLH in the control experiments (Fig. 5B). The posttest response to stimulation of the trained site (69.3 ± 8%) was significantly decremented compared with the pretest response (p < 0.02; n = 10). In contrast, there was no change in the gill withdrawal to untrained-site stimulation (posttest response, 112.5 ± 14.1%; p > 0.6 for the comparison with the pretest response to untrained-site stimulation). Importantly, the difference between the posttest response to trained-site stimulation in the APV and control experiments was significant (p < 0.02).
In the above experiments, the APV was present during the pretests and posttests, as well as during training. To test whether our results would differ if the NMDA receptor antagonist were present only during training, we used a modified experimental protocol. This protocol was the same as that used for the DNQX experiments (below). We found that APV also blocked LLH when it was present in the abdominal ganglion only during training. The mean posttest response to stimulation of the trained site after training in APV did not differ significantly from the pretest response (posttest response, 120 ± 20.7%; p > 0.4 for the comparison with the pretest response; one-sample t test; n = 4; data not shown), nor was the mean posttest response to stimulation of the untrained site in the APV experiments significantly different from the pretest response (posttest response, 106 ± 11.4%; p > 0.5 for the comparison with the pretest response). There was also no significant difference between the mean posttest responses to trained-site and untrained-site stimulation after training in APV (p > 0.5; paired t test). In contrast, in control experiments performed on animals from the same shipments that were used for the APV experiments, we observed LLH of the trained-site response. The mean posttest response to stimulation of the trained site after training in ASW was significantly habituated (posttest response, 47.5 ± 6.9%; p < 0.005 for the comparison with the pretest response; one-sample t test; n = 4; data not shown). Furthermore, the mean posttest response to trained-site stimulation was significantly less than that to untrained-site stimulation after training in ASW (p < 0.05; paired t test). The mean posttest response to stimulation of the untrained site in the ASW experiments was not significantly different from the pretest response (posttest response, 105.8 ± 16.7%; p > 0.7 for the comparison with the pretest response to untrained-site stimulation). Significantly, the posttest response to trained-site stimulation was more habituated after training in ASW than after training in APV (p < 0.03). These results support the conclusion from our other experiments involving APV (Fig. 5) that the presence of an NMDA receptor antagonist during training disrupts LLH.
Long-lasting habituation of gill withdrawal requires activation of AMPA-type receptors
It has been long believed that homosynaptic depression of transmission between the sensory and motor neurons in the abdominal ganglion, an exclusively presynaptic form of plasticity (Castellucci and Kandel, 1974; Armitage and Siegelbaum, 1998), is a cellular mechanism of habituation of the withdrawal reflex (Kandel et al., 1975). Armitage and Siegelbaum (1998) have shown that application of DNQX, an antagonist of AMPA-type receptors in Aplysia motor neurons (Dale and Kandel, 1993; Chitwood et al., 2001), does not disrupt short-term homosynaptic depression, although DNQX blocks transmission at the sensorimotor synapse. They also found that repeated application of glutamate, the sensory neuron transmitter (Dale and Kandel, 1993; Levenson et al., 2000), to the motor neuron did not depress the synaptically evoked sensorimotor EPSP. On the basis of their results Armitage and Siegelbaum concluded that activation of postsynaptic glutamate receptors was neither necessary nor sufficient for the induction of homosynaptic depression of sensorimotor synapses.
We tested whether activation of postsynaptic AMPA-type receptors was necessary for the induction of LLH. We infused DNQX (500 μm in ASW with 0.2% DMSO) into the abdominal ganglion during habituation training. To give adequate time for the drug to be washed in and washed out of the abdominal ganglion, we modified our basic experimental protocol. In these experiments, there was a 1 hr rest period between the end of the pretests and the onset of habituation training. DNQX was introduced into the abdominal artery immediately after the pretests and was present throughout the training. After the last block of training, there was a 2 hr rest period during which the DNQX was washed out of the abdominal ganglion (using ASW with 0.2% DMSO). Another modification in the protocol was that there were two pretests to both the trained and untrained sites (the pretests were administered alternately to the two sites at 5 min intervals). Each of the sites also received two posttests (also administered alternately to the sites at 5 min intervals). The presence of DNQX during habituation training blocked the induction of LLH (Fig. 6A). The mean posttest response of the gill to trained-site stimulation (108.4 ± 10.6%) was not significantly different from the pretest response (p > 0.6; n = 6). Furthermore, the mean posttest response to trained-site stimulation was not significantly different from that to untrained-site stimulation (140.6 ± 23.5%; p > 0.2). Control experiments were performed using a protocol identical to that used in the DNQX experiments, as well as animals from the same shipments. The abdominal artery was infused with ASW and 0.2% DMSO throughout the control experiments. The habituation training induced LLH of gill withdrawal in the control experiments (Fig. 6B). The mean posttest response to stimulation of the trained site was 61.9 ± 3.2% (p < 0.04; n = 6). Also, the mean posttest response to trained-site stimulation was significantly less than the mean posttest response to untrained-site stimulation (113.2 ± 12.2%; p < 0.04) in the control experiments. Finally, the mean posttest response to trained-site stimulation was significantly greater in the DNQX experiments than in the control experiments (p < 0.003).
Notice that, although the DNQX significantly reduced gill withdrawal to the training stimuli, the blockade of the response was incomplete (Fig. 6A). The concentration of DNQX that we used was approximately an order of magnitude greater than that shown in previous studies to produce a >90% blockade of synaptic transmission at sensorimotor synapses (Dale and Kandel, 1993; Armitage and Siegelbaum, 1998). The reduced gill responses observed during training in the presence of DNQX might reflect the contribution of peripheral sensorimotor synapses to the reflex (Peretz and Estes, 1974; Bailey et al., 1979; Frost et al., 1997). Such synapses would not have been affected by the DNQX, because the infusion of this drug was limited to the abdominal artery, which supplies the abdominal ganglion. Early work indicated that the peripheral nervous system alone can support habituation of gill withdrawal in Aplysia (Peretz, 1970; Lukowiak and Jacklet, 1972; Peretz and Moller, 1974; Lukowiak and Jacklet, 1975; Peretz et al., 1976). If the remnant of the withdrawal response apparent during training in DNQX is indeed mediated by the peripheral nervous system, our data confirm that the isolated peripheral nervous system can mediate short-term habituation (STH) of gill withdrawal, at least to some extent. An alternative explanation for the inability of the DNQX infusion to completely eliminate gill withdrawal in our experiments is that the drug did not have access to all of the sensorimotor synapses in the abdominal ganglion.
In all of the above within-preparation experiments, we excluded from our data analysis those preparations in which the posttest response to stimulation of the untrained siphon site was at least 80% of the pretest response (see Materials and Methods). We did this because we wanted to exclude results from preparations that had deteriorated. However, because it could be argued that we thereby biased our results, we reanalyzed all of our within-preparation data after including the data from the excluded preparations (n = 6). Adding the excluded preparations to our data did not alter any of our main results.
The role of protein synthesis in LLH in Aplysia
The present data demonstrate that prolonged habituation of the withdrawal reflex in Aplysia differs from STH in its requirement for protein synthesis. The lack of a requirement by STH for protein synthesis agrees with results from a previous study (Schwartz et al., 1971). Our study, however, represents the first demonstration that prolonged habituation of the withdrawal reflex requires protein synthesis. A requirement for protein synthesis has been also been observed for LTH of reflexive responses in the nematode worm Caenorhabditis elegans (Rose and Rankin, 2001; Rose et al., 2002) and the crab Chasmagnathus (Pedreira et al., 1996; Hermitte et al., 1999), as well as for LTH of the responsiveness to distress cries in mice (Squire and Becker, 1975).
We do not yet know whether LLH depends on RNA synthesis, as well as protein synthesis. An intermediate form of sensitization has been described recently in Aplysia (Sutton et al., 2001). Intermediate sensitization differs mechanistically from short-term sensitization in its dependence on protein synthesis. Intermediate sensitization differs from long-term sensitization (Frost et al., 1985; Castellucci et al., 1989) in that long-term sensitization requires RNA synthesis, whereas the intermediate form does not. Possibly, LLH represents an intermediate phase of habituation, analogous to intermediate sensitization.
Implications of site specificity of LLH for localizing the cellular site of plasticity
Because LLH of siphon-elicited gill withdrawal exhibits site specificity, the behavioral decrement cannot be ascribed to motor or muscle fatigue or to a gradual deterioration in the health of the preparations. The demonstration of site specificity also places certain limits on the locus of the cellular changes that underlie LLH. For example, LLH cannot be attributable to a general, state-related change within the CNS. Furthermore, if LLH is attributable, at least partly, to enhanced inhibitory input to the withdrawal circuitry, then the inhibitory input must be somewhat specific with respect to sensorimotor pathways within the abdominal ganglion.
Stopfer et al. (1996) have shown previously that STH and LTH (lasting 24 hr) of the tail-elicited siphon-withdrawal reflex exhibit site specificity. Stopfer et al. argued that site-specific habituation in their experiments proved that "the cellular sites of plasticity must exist upstream from the motor neurons," because the siphon motor neurons receive common interneuronal input from both sides of the tail. However, the motor neurons might well be a site of plasticity in our experiments. Site specificity of habituation can be maintained with the cellular locus of plasticity in the motor neurons if the learning-related changes are restricted to specific postsynaptic sites. For example, both NMDA receptor-dependent long-term potentiation and LTD in the mammalian CNS exhibit input specificity, although these forms of plasticity are induced and, at least in part, expressed postsynaptically (Bliss and Collingridge, 1993; Bear and Linden, 2001).
Stimuli that induce habituation of the gill-withdrawal reflex appear to activate sensitizing cellular processes as well
In 1970, Groves and Thompson proposed the "dual process" theory of habituation. According to this theory, stimuli that induce habituation in a particular stimulus-response pathway also recruit an independent, incremental (sensitizing) process that increases the state of excitation in an organism. The final response strength in the Groves and Thompson model is determined by the net outcome of the countervailing processes of habituation and sensitization. Our data provide support for the dual-process theory of habituation. Treatments that blocked LLH (inhibition of protein synthesis, inhibition of protein phosphatase activity, and antagonism of NMDA-type receptors) also revealed the effects of a sensitizing process elicited by habituation training. This sensitizing process is apparent in the tendency of the withdrawal responses to trained-site stimulation to increase in strength over the course of habituation training (Figs. 3B, 4A, 5A).
Homosynaptic, exclusively presynaptic, depression of the sensorimotor synapses is not the mechanism of LLH in Aplysia
The data from the present study indicate that homosynaptic depression of sensorimotor synapses, an exclusively presynaptic form of plasticity (Castellucci and Kandel, 1974; Armitage and Siegelbaum, 1998; Royer et al., 2000; Gover et al., 2002), is insufficient for LLH. In most of the experiments that used pharmacological blocking agents, there was a dissociation between the effect of the agents on LLH and their effect on STH. Specifically, neither anisomycin (Fig. 3B), OA (Fig. 4A), nor APV (Fig. 5A) blocked STH, yet each blocked LTH. Given that STH is mediated primarily by homosynaptic depression (Frost et al., 1997; Antonov et al., 1999), our results imply that homosynaptic depression of sensorimotor synapses is insufficient for the induction of prolonged habituation. The DNQX experiment provides compelling evidence that homosynaptic depression is not the mechanism of LLH. This is because application of DNQX to the sensorimotor synapse does not alter short-term homosynaptic depression (Armitage and Siegelbaum, 1998), whereas application of DNQX to the abdominal ganglion blocked LLH (Fig. 6A).
An alternative explanation for our DNQX result is that the long-term cellular change that underlies LLH occurs exclusively, or mostly, at synapses between interneurons and motor neurons (below). According to this hypothesis, the DNQX blocked synaptic transmission at the sensory neuron-to-interneuron synapses. This, in turn, prevented activation of the interneurons, thereby disrupting long-term plastic changes at the interneuron-to-motor neuron synapses. Although this explanation cannot be ruled out by our data, it seems unlikely given that LTH of gill withdrawal is accompanied by significant long-term depression of sensorimotor synapses in the abdominal ganglion (Castellucci et al., 1978).
The role of postsynaptic glutamate receptor activation in LLH
Our data indicate that activation of postsynaptic glutamate receptors (Dale and Kandel, 1993; Trudeau and Castellucci, 1993; Lin and Glanzman, 1994; Conrad et al., 1999; Yung et al., 2002) is critical for the induction of LLH. Antonov et al. (2003) have reported recently that high concentrations of DNQX can significantly reduce the NMDA-type receptor component, as well as the AMPA-type receptor component, of the sensorimotor EPSP (Antonov et al., 2003). Possibly, the critical action of DNQX in our experiments was on the NMDA type rather than the AMPA type. Future experiments with more selective AMPA receptor antagonists can resolve this issue. Interestingly, a study of LTH of the tap withdrawal response in C. elegans has found that mutant worms lacking a homolog of the vertebrate AMPA receptor [glr-1 (glutamate receptor 1) mutants] exhibit normal STH to spaced blocks of taps but do not show LTH (Rose et al., 2001).
Does an LTD-related synaptic mechanism mediate LLH in Aplysia?
The cellular processes essential for LLH induction are reminiscent of those necessary for the induction of homosynaptic LTD of synapses in the mammalian hippocampus and cortex (Mulkey and Malenka, 1992; Mulkey et al., 1993; Kirkwood and Bear, 1994). According to a current model of LTD (Bear and Linden, 2001), low-frequency stimulation (LFS) of afferent inputs causes modest activation of postsynaptic NMDA receptors, resulting in a moderate influx of Ca2+ into the dendrites of the postsynaptic cell. This postsynaptic influx of Ca2+, in turn, activates calcineurin (PP2B), which releases PP1 from inhibition, thereby permitting PP1 to become active. Activation of these protein phosphatases leads to long-term synaptic weakening. The precise pathways by which activation of PP2B and PP1 produces LTD are not completely understood, but it is believed that the protein phosphatase cascade causes the endocytosis of AMPA receptors, thereby resulting in fewer receptors in the postsynaptic membrane (Carroll et al., 2001).
An LTD-like form of synaptic plasticity has been reported previously for Aplysia sensorimotor synapses (Lin and Glanzman, 1996). Like homosynaptic LTD of hippocampal synapses (Mulkey and Malenka, 1992), LTD of sensorimotor synapses depends on a postsynaptic rise in Ca2+ because it is blocked by postsynaptic injection of the rapid Ca2+ chelator BAPTA before the onset of LFS. Whether this form of LTD of Aplysia synapses also resembles LTD of mammalian synapses in its requirement for activation of protein phosphatases and NMDA receptors is not yet known.
The potential role of interneurons in LLH of siphon-elicited gill withdrawal
The present data do not rule out contributions from changes in interneuronal pathways. Indeed, the previous failure to obtain long-term (24 hr) depression of in vitro sensorimotor synapses using various patterns of homosynaptic stimulation, together with the demonstration that repeated applications of the inhibitory peptide FMRFamide (Piomelli et al., 1987) can induce long-term depression (Montarolo et al., 1988), suggest that contributions from heterosynaptic mechanisms might play a role in LLH. A potential link between our finding that inhibition of protein phosphatase activity can block the induction of LLH (Fig. 4A) and a possible role for FMRFamide in LLH is provided by evidence that at least some neuronal actions of FMRFamide within the CNS of Aplysia are mediated by activation of protein phosphatases (Sweatt et al., 1989; Ichinose and Byrne, 1991).
Habituation of the withdrawal reflex in Aplysia is frequently regarded as one of the simplest forms of learning with respect to its underlying cellular mechanisms. However, the present data demonstrate that our current understanding of habituation of the withdrawal reflex is very incomplete. Postsynaptic mechanisms play a far more important role in habituation than has been heretofore appreciated. In particular, our data raise the possibility that an LTD-like synaptic mechanism may mediate this simple form of invertebrate learning.
This work was supported by National Science Foundation Grant IBN-9808930 and National Institutes of Health Grant NS29563 (D.L.G.). We thank Drs. Frank Krasne, Marc Klein, Tom O'Dell, and Adam Roberts for their helpful comments on this manuscript.
Correspondence should be addressed to Dr. David L. Glanzman, Gonda (Goldschmied) Neuroscience and Genetics Research Center, 695 Young Drive South, Box 951761, University of California, Los Angeles, Los Angeles, CA 90095-1761. E-mail: dglanzman{at}physci.ucla.edu.
Copyright © 2003 Society for Neuroscience 0270-6474/03/239585-10$15.00/0


text: Working while remote NT communities are locked downWritten on the 8 December 2021 by Kelly Gulliver The NT recently experienced their first outbreak of COVID-19 in a number of remote communities surrounding Katherine— otherwise known as The Big Rivers region. The communities affected were Robinson River, Binjari, Rockhole and Lajamanu. Binjari and Rockhole went into 'hard lockdown' for over a week, meaning residents where not permitted to leave their communities or return home if they were in Katherine at the time. This brought about new challenges for the NT Welfare and Homelessness Group made up of NGO's and NT Government departments who met daily via zoom for over three weeks to collaborate on strategies and implement contingencies to keep people safe, housed and fed during the lockdown. The smaller communities such as Binjari and Rockhole do not have their own food supply and residents usually travel into Katherine via car, taxi or the bush bus to buy supplies. CCNT Financial Wellbeing and Capability and Housing Support teams worked with foodbank to supply over 1000 food hampers in total during the lockdown and joined forces with the NTG critical goods team and defence to distribute emergency relief hampers to those communities in hard lockdown. The communities' houses have a pre-paid electricity system that requires the purchase of power cards to keep electricity running in the house. The CCNT team used the DSS funded ER to purchase $2000 worth of power cards to be distributed to Robinson river, Binjari and Rockhole households. The power cards were well received due to power consumption being up by about 20% with the wet season looming and large families being forced to stay indoors to follow CHO orders. In Katherine, there is a high population of people sleeping rough and living in overcrowded housing so as more and more exposure sites were announced, close contacts needed support to be transported to the Centre for National Resilience at Howard Springs for quarantine. Casual contacts required NGO support to access safe accommodation that allowed them to isolate and follow the direction of the CHO. The NTG set up brokerage for this support which was administered by CCNT and Salvation Army in Katherine. In total over 220 people were placed into local motel accommodation during the lockdown and lockout periods with this much needed Government funding. The majority of clients vacated on 8 December to return to their usual place of residence. Some communities have their own land council restrictions above and beyond the CHO direction which means they cannot return home until 17 December. There was a lot of learnings from this experience about how a lockdown looks in a remote setting, as the needs of each community is unique and varies significantly to that of a more urban setting. The people on the ground worked together tremendously to get the job done. Author:Kelly Gulliver |

subset: commoncrawl


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The 8-8 Stargate is a cosmic portal of tremendous power that opened on July 26th (the first day of the Egyptian calendar) and closes August 12th.
Its energy transfer to Earth peaks on August 8th when Sirius rises in the east and is visible on the horizon just before the sun rises. On the 8-8 Earth aligns with both Sirius and our Sun, a very powerful alignment charged with the Heart of the Lion.
Leo governs the heart and the solar plexus so the 8-8 Stargate gives us an extraordinary opportunity to amplify and maximize our Spiritual Light and Inner Power!
We will be doing a Global Meditation at the peak moment of this incredible Ascension Portal. At 8:08 am your local time begin the Lions Gate guided meditation (link below) followed by a brief Flower of Life Meditation immediately afterwards to stabilize the Earth Grids and global situation (link below).
Important Highlights of the 8-8 Lions Gate Portal
- Tune into the Higher Mind, Higher Heart, Higher Wisdom
- Opportunity to harness a limitless supply of Power, Infinity and Immortality
- We can use it to Jump Timelines since the 8-8 Lions Gate is a TIME PORTAL
- It supercharges our Consciousness
- Allows us to Reconnect with the Feminine Divine and the Mysteries of the Goddess
- The Empowerment Gateway – Fully Step Into Your Personal Power
- Connect with the Great Central Sun and Shine With the Light Of A Thousand Suns
- Discharges powerful ascension light codes and is an Accelerated Ascension Portal used by the ancients
TIME August 8th, 2020 8:08am your local time (it is recommended to use the guided audio below for this meditation)
1. Use your own technique to bring you to a relaxed state of consciousness.
2. See a large Violet Flame enveloping your body and your home. Ask St. Germain to amplify this flame 1,000 times its normal strength and transmute any darkness both during and after the meditation.
3. Turn off all identification with linear time and expand your energy field out – first to a 100 mile radius, then to global level, galactic level and finally universal level where you merge with the divine light of omni-presence.
4. Envision a large pillar of extremely bright blue Light emanating from Sirius and inside of this pillar are myriads of shimmering golden stars. See this pillar of dazzling light stream down into your body and anchor into the center of the Earth. Keep this pillar active throughout this cosmic journey. Call in your I Am presence, your oversoul, positive Star Beings from Sirius and any other light beings you work with.
5. Concentrate on opening your 3rd Eye Chakra / Pineal Gland. You will see a swirling violet light and what may appear as a patch of blue sky as the portal opens and becomes wider. This is the built-in travel mechanism you will use to get to the higher dimension and the 5D Earth Grid.
6. See yourself leaping off the old 3D Matrix Grid……..see it collapsing and disintegrating. Watch every last vestige of darkness being vaporized beneath your feet as you travel upwards at warp speed. You can see the new fifth dimensional crystalline Earth Grid shimmering like a jewel in the sky! As you gently land on the surface of this dazzling new world make this decree "I have successfully transferred my Consciousness to the crystalline 5D Earth Grid and from this moment on, THIS is the world I live in and experience on every level!!!
7. You have entered the realm of the Goddess (our Divine Mother). Feel her tenderness filling your heart and her gentle embrace soothing your spirit. Feel the Goddess supporting you and granting the deepest desires of your heart. Feel the ecstasy of her love lifting you up and giving you wings to FLY!
8. Now spend a few minutes seeing our New 5D World completely built and thriving – in its most glorious form! Envision the New Cities of Light in grand detail and the various advanced healing and replication technologies being accessible to all. See the flourishing of creative arts all around the planet.
Take a stroll in your city or community and marvel at all of the beauty surrounding you – paint in as much detail as your wild imagination can conjure. Envision using all your newly restored superpowers and metaphysical abilities. Take a few minutes to explore 5D Earth and Create the Life Of Your Dreams!
9. Finally, go into the silence for a few minutes to receive important spiritual downloads from the Sirius Star System that you need at this time.
Follow this meditation by the Flower of Life Meditation: (Instructions and Guided Audio in Different Languages at Link)
The 8-8 is a day when we can more securely anchor our consciousness onto the Positive Ascension Timeline and Leap off the 3D Matrix Timeline altogether, so we don't want to miss this opportunity!
Although our Triple Eclipse meditations averted many worse scenarios, there will be some very volatile astrological alignments starting mid-August which could stir up anger in the collective, so let's participate in this Meditation to continue stabilizing the global situation and ensure the best smoothest outcome over the coming weeks and months!
Even though 8-8 is called "The Lions Gate" and seems to carry strong masculine overtones, it is actually a "Stargate of the Goddess" that streams in divine feminine energy which restores harmony and balance on our planet.
The Goddess is the feminine aspect of Source Creator and her presence was greatly suppressed on Earth many millennia ago, so this portal can help us revive her presence and reconnect with her loving healing energy.
Since we are in the final showdown between Light and Dark here on Earth, goddess energy has never been more needed than it is right now! So we will want to strongly connect with the Goddess / Feminine Divine throughout this entire cosmic window and especially on August 8th to bring stability to the global situation and ensure peace will reign in our domain!!
The veil between the material and spiritual realms are very thin on 8-8 so it will be much easier to connect with the Goddess, Source Creator and angelic beings on that day.
Just as our physical sun is considered to be the giver of life, Sirius has long been viewed as our spiritual life-giver as it illuminates the inner planes of existence.
SIRIUS is the brightest star in our night sky. It is twice as large in mass as our sun and 26 times more luminous – quite a magnificent star indeed!
Because it is such a brilliant luminary, Sirius is considered to be a broadcasting station of spiritual wisdom. These light transmissions that are now flooding the Earth are packed full of the codes of Spiritual Mastery, Evolution and Ascension.
These diamond light codes originating from the Great Central Sun can activate our DNA, strengthen our Aura (our personal energy field), prime our Pineal Gland and fuel our Quantum Leaping abilities!!
Sirius is often referred to as the "Dog" star because it is found in the Canis Major constellation which is where we get the word "canine" from. One of the oldest gods of Egypt was Anubis and he is portrayed as having the head of a jackal.
A major component of the sacred knowledge that was given to the Atlanteans by the far more evolved Cosmic Central Race, was connecting with and embodying the goddess presence which in turn brought peace, harmony and stability to the whole of society. This is why the goddess was so revered in many ancient cultures and why temples were erected in her honor.
In addition to the power of the astrological alignments during the Lions Gate, we also have the added mathematical manifestation power embedded in the numbers 88.
8 is the number of Abundance, Wealth and Personal Success. The two equal circles stacked atop each other signify balance – 8 brings balance between the spiritual and material realms.
The infinity symbol is the number 8 lying on its side so the power of infinite potential lies within the 8!
88 is a universal code tied to our grandest dreams. When we express and share our innate gifts we align with the higher divine expression of ourselves and the Universe will support us every step of the way!
The 8 is a very fair number – it will dole back out to you in equal measure what you have sent out to others. If you have made many deposits in the spiritual bank of the higher realms for years, you can expect a very large payout soon (with interest).
Those who honor the light and follow the sacred path of love are about to be greatly rewarded. But the same 8-8 passage will spell disaster for those who operate on the tenets of darkness and oppression, such as those who created and run the Matrix. They will not fare so well during this extra potent Lions Gate.
So let's harness the immense manifestation power of this Stargate by quantum leaping through the 8-8 Portal, as within the #8 lie ALL POSSIBILITIES……To Infinity!!!
On this day, we can confidently take a total leap of faith, bounce off of this starlit trampoline and land among the stars of our New Reality, the grandest reality we can envision!
To Read Full Article: https://thevoice5d.blogspot.com/2020/07/8-8-lions-gate-ascension-portal.html
Disclaimer: We at Prepare for Change (PFC) bring you information that is not offered by the mainstream news, and therefore may seem controversial. The opinions, views, statements, and/or information we present are not necessarily promoted, endorsed, espoused, or agreed to by Prepare for Change, its leadership Council, members, those who work with PFC, or those who read its content. However, they are hopefully provocative. Please use discernment! Use logical thinking, your own intuition and your own connection with Source, Spirit and Natural Laws to help you determine what is true and what is not. By sharing information and seeding dialogue, it is our goal to raise consciousness and awareness of higher truths to free us from enslavement of the matrix in this material realm.


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WINDSOR, Colo. - The Town of Windsor Arts Commission announces its 2023 Art LIVE! event schedule that allows residents to watch professional artists create artwork during some of Windsor's biggest events.
Windsor residents are invited to attend and meet local professional artists, watch their creative process live, and learn more about community art plans in Windsor. These events will vary in location and time each month. Attendees will have the chance to receive exclusive ART LIVE! swag such as magnets, water bottles, and sunglasses, while supplies last.
The first event will happen during the Easter Eggstravaganza event at the Community Recreation Center, 250 N 11th St., on Friday, April 7, from 11 a.m. to 12 p.m. Artist Laura Grossett will be showcasing a printmaking demonstration and presenting the acrylic paint mural she painted inside the recreation center lobby.
Additional dates, locations, and details for Art LIVE! include:
Artist details are to be determined. Follow events on Facebook and Instagram @windsorprc, or by visiting, recreationliveshere.com/190/Public-Art.
For more information about the Town of Windsor Arts Commission, visit windsorgov.com/1316/Arts-Commission. To view art from local artists, events or exhibits, visit recreationliveshere.com/190/Public-Art.


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Youngstown Thermal in Talks with Potential Buyer
YOUNGSTOWN, Ohio – A company based in suburban Columbus is performing due diligence toward a possible purchase of Youngstown Thermal LLC's assets — an acquisition that could result in lower rates for customers, according to documents filed with the Mahoning County Court of Common Pleas.
SOBE Energy Solutions Inc., Dublin, has sent a non-binding letter of intent "for the proposed purchase of Youngstown Thermal" to the downtown district heating and cooling utility, according to the receiver's fourth report.
"The receiver determined that the prospective acquisition by SOBE would likely result in lower rates since SOBE would provide ancillary services which would offset the need for rate increases," the report said.
Youngstown Thermal was placed into receivership in August 2017 to restructure the failing business. The district energy system supplies steam heat and some cooling services to about 30 customers and 22 buildings downtown, including City Hall.
Reg Martin, the court-appointed receiver, could not be reached for comment.
Stephen Hubbard, CEO of SOBE, said it would be premature to comment on any agreement or purchase. "There is a lot of due diligence to be completed yet," he said.
According to its website, SOBE Energy uses waste-to-energy conversion technology that produces a clean, synthetic fuel gas that can be used directly in burners for process heating, or in gas turbines or reciprocating engines for electricity generation.
SOBE's letter of intent includes a proposal to the use biomass oil through Ensyn, a renewable energy company based in Canada, according to documents. The company supplies Youngstown Thermal with fuel oil that is converted from wood waste, which should reduce its energy costs by about $1 million this year, according to previous court filings by Youngstown Thermal. Ensyn provides at least 70% of Youngstown Thermal's fuel.
Still, the most recent receiver's report cautions that as of yet, no deal has been reached with SOBE and that the downtown utility should continue to pursue other options.
The most preferable option is for the city to acquire Youngstown Thermal, the report stated. Should this happen, Ensyn has indicted that it would construct a new renewable energy plant in the city that would create 400 construction jobs and 50 permanent positions.
Plus, a city-owned utility would ensure that heating and cooling rates remain reasonable for downtown customers, Martin said during a presentation last summer to Mayor Jamael Tito Brown and members of City Council. A sale to a for-profit corporation, he said, would likely result in higher rates for consumers.
The mayor stated last year that the city is not interested in operating a steam-heat plant, and would not consider taking it over.
Another option would be to create a cooperative among the central business district's customers to run the plant on North Avenue, the report stated.
"I think I'd be in favor of exploring that with the stakeholders," said Fourth Ward Councilman Mike Ray, chairman of city council's public utilities committee. "District steam makes sense if it's run properly."
However, Ray said that the administration would have to support the effort. "It's good to at least start the conversation," he said.
Nevertheless, the receiver "has executed non-disclosure agreements and exchanged confidential with two other potential purchasers for Youngstown Thermal," according to the most recent report.
Martin has said that maintaining lower rates for customers is key to signing up others to use the system. In September, the company received notice that the Youngstown Fire Department would discontinue using Youngstown Thermal's steam service, the report said.
"Unfortunately, the loss of even one customer has a negative impact on all other customers as it causes rates to increase," the report stated. "It is in the best interests of all customers as a whole that each individual customer continue to use Youngstown Thermal as their provider."
Copyright 2024 The Business Journal, Youngstown, Ohio.


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Let your garden take flight with the new Diggity Garden Bird Stake. This cleverly designed garden sculpture is a faux shovel head that has been painted and accessorized to look like a sweet little bird flying out of your garden bed.
Made of metal and patchwork bits in rustic colors, this looks like something you would buy at a country arts and crafts fair, but at half the price! Diggity Stakes are made of weather-resistant metals so they will withstand whatever Mother Nature throws their way, too. Tough and beautiful! You've got to love that.
The stake is over 3 feet tall, so you can put it mid-border and let it peek out from your shrubs for a cute element of surprise. Try it in a flower pot and let summer vines grow up the stake for a pretty welcome by the front door or garden gate.
Let this little bird bring happiness to your garden and order yours today!
Write Your Own Review


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You may not have noticed, but Bayern Munich appear to have updated their logo.
Liverpool to face Bayern at Audi Cup
An eagle-eyed Facebook user noticed the the lettering on the club's crest had changed in recent post sent out from the German champions' official account.
Bayern responded to the user, confirming that the logo had indeed been modified with a slightly different font and that the colours had been updated.
A graphic designer who is affiliated with the club, Daniel Nyari, commented on Twitter that the red is now warmer, the the blue is darker and that the angle Bavarian lozenges has changed
corresponding reds backgrounds. but the kit primary colors can vary & do for a lot of clubs pic.twitter.com/AuoIZddqzV
— Daniel Nyari (@danielnyari) June 16, 2017
Nyari also added that the changes were made due to global non-football competition, to consolidate brand tone and add more brand visibility.


text: Next-generation robots automate large-scale industrial processes Robotic or automated systems have their limitations when it comes to large-scale product life cycles. An EU initiative has designed a flexible and reconfigurable robotic system to accelerate maintenance and repair processes in large-scale products. Industrial Technologies © Thinkstock Automation in the aeronautics and construction industries has been hindered by the need to move sizeable and heavy structures as well as the risk of damaging automation equipment. These major issues have motivated inquiry into automated processes that will be flexible and cost effective. The EU-funded CABLEBOT (Parallel cable robotics for improving maintenance and logistics of large-scale products) project was uniquely designed to meet these needs. Researchers worked on developing a new generation of modular and reconfigurable robotic devices that are capable of performing different maintenance steps during the life-cycle stages of large-scale structures. To achieve automation and flexibility in large-scale maintenance, team members used novel software tools to design parallel cable robots. These robotic solutions meet the needs of products that have differing dimensions and features. Simulations were carried out to verify the robotic device's operation in environments with large-scale structures. Software tools and methods were designed to simulate the different requirements of each life-cycle stage. Project partners also developed control algorithms and strategies to operate the cable robots based on industrial requirements. Lastly, two industrial use cases tested the software tools and cable robots developed, and demonstrated the efficiency of the control algorithms and strategies. They also validated the feasibility of employing cable robots in aircraft maintenance and in handling and assembling construction beams. CABLEBOT proved the value of parallel cable robots as a cost-effective and flexible alternative to conventional industrial robots in automating large-scale processes, particularly for the aerospace and construction industries. As a result, the service life of products will be extended while reducing their overall life-cycle costs. Last June, Euronews visited the facilities of TECNALIA in Montpellier where the prototype COGIRO is installed. COGIRO is Europe's biggest Cable-Driven Parallel Robot and was developed by CNRS-LIRMM and TECNALIA. The video produced by Euronews can be seen here. Keywords Robots, industrial processes, large-scale product, product life cycles, parallel cable robotics, cable driven robots


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Data Reviewer のライセンスで利用可能。
The latest version of ArcGIS Data Reviewer Server can be installed without a user interface by running the setup.exe from the command prompt. You can also install ArcGIS Data Reviewer Server silently in an ArcGIS Server multi-machine deployment.
Upgrading to 10.8.1? For more information, see Upgrading existing ArcGIS Data Reviewer Server sites.
Installation prerequisites include the following:
- Authorizing the software for the machine on which you will install ArcGIS Data Reviewer Server.
- Verify that your site meets the system requirements.
- Log in as a user with administrative privileges.
In certain scenarios, the ArcGIS Server account can be set to Local System when ArcGIS Server is installed silently. In these scenarios, it is recommended that you change the ArcGIS Server account by running the Configure ArcGIS Server Account utility.
- Open a command prompt.
- Execute the following command:
<path to ArcGIS Data Reviewer for Server>\setup.exe /qb
For example:
\\machinename\ArcGIS_DataReviewer_for_Server\setup.exe /qb
To install ArcGIS Data Reviewer Server in an ArcGIS Server multi-machine deployment, run this command on each machine in the site.
Proceed to Configuring ArcGIS Data Reviewer Server to set up your server.


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SOURCE: Bay Area Wine Project
CHICAGO, IL–(Marketwire – October 6, 2010) – More than 300 acclaimed pinots will be poured at the 4th Annual Pinot Days Chicago, a pinot noir wine event that takes place November 11th-November 13th, 2010. Pinot Days, www.pinotdays.com, connects wine enthusiasts directly with winemakers creating a unique opportunity for new and seasoned wine lovers to experience, enjoy and deepen their knowledge of pinot noir. Pinot Days includes intimate wine gatherings and tastings, winemaker dinners, and retail store tastings. Pinot Days culminates with the Pinot Days Grand Festival at historical Navy Pier in Chicago, IL, which gives wine enthusiasts the opportunity to experience and enjoy some of the world's best pinot noir and interact directly with the more than 70 acclaimed winemakers who produce them. Event tickets and detailed information about Pinot Days can be purchased at www.pinotdays.com.
Founded by The Bay Area Wine Project with the ultimate goal of advancing the profile of pinot noir, Pinot Days is a three-day celebration of Pinot Noir and its most brilliant winemakers, the vineyards, the grapes, and the often-bewitching process of urging the noble pinot grape into great wine. At Pinot Days, the winemakers and winery owners do the pouring, creating a uniquely engaging experience where wine enthusiasts can interact, ask questions and learn the stories behind what goes into making great pinot noir.
Pinot Days founder Steven Rigisich believes 2010 may be the best year in Pinot Days Chicago's 4-year history. She explained, "Pinot noir is at its best, with quality and breadth of style increasing with every vintage. The winemakers attending this year's Grand Festival are among the best in the country. We expect this year to be the ultimate celebration of the 'noble grape.'"
Pinot Days events for 2010 will include a hosted Winemakers "Table Hop" Dinner at Uncommon Ground, and a "Pinot 101" with Sommelier Brian Duncan, held at the renowned Bin 36 restaurant, featuring 16 notable pinot noir producers including Rod Berglund of Joseph Swan, Ed Kurtzman of August West, and Wes Hagen of Clos Pepe.
Each year, Pinot Days conducts a large-scale annual charity auction at the San Francisco event, the proceeds of which go to one IOCC charity. This year's Pinot Days Charity Auction Beneficiary is the IOCC's Prevention of Mother to Child HIV/AIDS Transmission (PMTCT) activities currently being undertaken in Ethiopia. 92% of the monies raised for this cause will go directly to providing food to people in the greatest need. Pinot Days Annual Charity Auction has raised over $91K to date for various beneficiaries.
Pinot Days is an event that connects wine enthusiasts directly with winemakers creating a unique opportunity for new and seasoned wine lovers to experience, enjoy and deepen their knowledge of pinot noir. Produced by the Bay Area Wine Project, a non-profit organization dedicated to the promotion of pinot noir, Pinot Days was created by Steven and Lisa Rigisich out of their love for the "the noble grape." The Bay Area Wine Project holds annual events in San Francisco, Chicago, and Southern California. Pinotdays.com provides information about and access to wine-related events, wine education opportunities and wine trivia — including articles about outstanding wine personalities, winery profiles, reviews of wine events, restaurant wine lists, formal tastings and more. For more information visit www.pinotdays.com.
Pinot Days/Bay Area Wine Project
Lisa Rigisich
[email protected]
Click here to see all recent news from this company


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The fact is that a fully healthy, or even mostly healthy Packer team wins this game rather easily. Every Aaron Rodgers interception would have been a big gain or touchdown if he were healthy, and if Davante Adams wasn't knocked out on a despicable blindside hit by Thomas Davis, the Packers would have completed the comeback. Instead, the banged up defense couldn't stop Christian McCaffrey in the passing game or Cam Newton scrambling, and the offense just didn't have enough to keep up.
Chip On His Shoulder
I do not believe Brett Hundley could have won this game, however it was clear that Aaron Rodgers was impacted by his broken collarbone as his throws lacked zip, and more importantly, distance. Rodgers made smart reads as he always does, but his arm simply couldn't cooperate, which led to an unusually high number of turnovers as well as several big incompletions on out patterns. This was clearly a surprise to Rodgers as he attempted many throws he simply couldn't make, and failed to adjust to his limitations with shorter throws or more frequent run calls.
All interceptions are bad, but every one of Rodgers created a huge swing in the game. His first likely should have been a big gain to Davante Adams, who had beaten his man, and his second, a deep shot to Randall Cobb, probably should have resulted in a touchdown.
Jordy Nelson also managed to get behind his man on the third pick of the game, but by the time the ball arrived a few yards short that huge opening became double coverage. Until he is fully healed, Rodgers needs to play to his strengths and keep his throws in the short-to medium range.
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That said, even with all of the turnovers, the Packers still had a very real shot to tie the game in the waning minutes, and they likely would have done so if Davante Adams was still on the field. The incredible cheap shot by linebacker Thomas Davis that landed Adams in the concussion protocol, but did not even result in an ejection for Davis, completely changed the game. Adams, who caught five of seven targets for 57 yards and a touchdown before exiting, dominated the Carolina secondary. He was open on nearly every play, and contributed yards after the catch consistently. Adams would likely have been the target on the final slant pass of the game, which was fumbled away by Geronimo Allison, and based on the spacing on that play there is a good chance that Adams would have taken it all the way.
There's an even better chance that the Packers would have simply produced enough offense to avoid having to recover an onside kick in the first place.
Carolina played well, but they also received several breaks including a ridiculous call on a touchdown pass to Damiere Byrd.
The play was originally ruled incomplete as Byrd landed half out of bounds, however the replay official inexplicably overturned the call, ruling that half of Byrd's posterior landed in bounds first. This is simply ridiculous for a number of reasons, and even the on-field official was unable, or unwilling to provide any rationale, stating that Byrd was "down by contact in the end zone." It is possible the the Panthers would have scored on the this drive anyway, but it's more likely that they would have settled for a field goal try and in any case, awarding a touchdown was unfair.
Given the state of the defense, the offense needed to come through with more than 24 points, and it simply wasn't in the cards, and with their playoff hopes dashed, they can begin thinking about the offseason, including how exactly they plan to deal with their defense.
The Capers Conundrum
It is well established that the most important part of any defense is the pass rush, and when you look to understand the issues the Packers have, it's almost always tied up in pass rush. While the secondary has suffered injuries to key starters for two consecutive seasons, that situation would have been mitigated if the team possessed a consistent pass rush. The best gift you can give a secondary, especially an inexperienced or bad secondary, is time, and a consistent pass rush from your front four is crucial. Capers' defenses specialize in generating pressure from blitz packages, which leads to a lack of consistency even if it does generate sacks. The result is that when a blitz doesn't get home, the secondary is hung out to dry, and that was frequently the case against the very mobile Cam Newton.
It is far too easy for opposing teams to neutralize Nick Perry and Clay Matthews, and while Mike Daniels does create some interior pressure, the sum of these parts is lacking. Matthews may have actually changed the game had he not fallen down on more than one occasion.
The bottom line is that Capers, Thompson, luck and the players themselves all share some blame for the Packers' defensive performance. And given the number of injuries and turnovers by the offense, it wasn't even that bad of a performance. That said, injuries are the norm in the NFL, and Capers' system does not cope well with them. It is in some ways a very balanced system, but that balance comes at the cost of specialization upfront. This isn't the best game to judge the defense on, but the edge rushers had a few chances to steal this one for the team by getting to Newton, or at least speeding up his internal clock. Instead a stretched secondary was unable to deal with tow of the more talented interior receiving threats in Greg Olsen and Christian McCaffrey along with the wideouts, and Newton had ample time to make simple throws to his run-after-catch specialists.


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- Pink - $ 15.99
Returns Policy
We try to avoid there being any issues or reasons why you might want to return an item from our shop but due to the handmade nature and being a small business we do have a policy of final sales on handmade items. So please, ask us whatever questions you may have up front. We're happy to help and get you the perfect item(s).If, for some reason, you do decide you want a different item or color we are happy to work with you to do an exchange but the item MUST be in brand new condition. Buyer is responsible for shipping charges and must place a new order for replacement.
Although we like to consider that we never make mistakes, but sometimes we do, and an item may be damaged. We'll make it right so please let us know by contacting us via email at firstname.lastname@example.com.
In the case we could not come to an agreement of an exchange and a refund has been decided, you should expect to receive your refund within four weeks of giving your package to the return shipper, however, in many cases you will receive a refund more quickly. This time period includes the transit time for us to receive your return from the shipper (5 to 10 business days), the time it takes us to process your return once we receive it (3 to 5 business days), and the time it takes your bank to process our refund request (5 to 10 business days).
You're order is shipping from the southern most State in the USA! Key West, Florida is only 90 miles from Cuba. Please allow 1 extra day for shipping times. Key West is a small 4 mile by 2 mile Island, and it takes time to get all that mail to the mainland.We can ship to virtually any address in the world. Note that there are restrictions on some products, and some products cannot be shipped to international destinations.
When you place an order, we will estimate shipping and delivery dates for you based on the availability of your items and the shipping options you choose. Depending on the shipping provider you choose, shipping date estimates may appear on the shipping quotes page.
Please also note that the shipping rates for many items we sell are weight-based. The weight of any such item can be found on its detail page. To reflect the policies of the shipping companies we use, all weights will be rounded up to the next full pound.
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PLEASE NOTE: We are NOT responsible for lost, stolen or undelivered packages. If you want to purchase insurance you may do so by contacting us prior to ordering. (Priority shipping automatically comes with $50 insurance) ALL LOST, STOLEN OR RETURNED packages paid without insurance are the responsibility of the buyer. USPS failures are not our fault or a reflection on my level of dedication to our customers. In any case, please contact me first with any issue.
International orders are averaging 10-14 business days for delivery. This depends on your country as well. We cannot control customs delays and we are not responsible for lost or stolen packages. If you want to add insurance to your order please contact us prior to purchasing. (Priority shipping automatically comes with $50 insurance)
Bring out your inner wild flower and don't hold back. Be a flower child, pop star, or queen of your own personal fashion show with our Buttercup Body Jewels. With a peel and stick application, you can be ready to go in seconds. A perfect accessory for festivals, theme parties, raves or just wearing at home while baking cookies or watching Netflix…whatever floats your boat! Make sure to pair your Pasties with matching Stickers for a full flowerlicious look.
- Hypoallergenic.
- Waterproof and durable.
- Blingy stickers to satisfy your sparkle craving.
- Wearable in many different ways, use some imagination and go wild.
- Reusable with body glue.
- Ready to go, just peel and stick.
- Measures 4 inches by 2.25 inches.


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Second driver arrested for fatal Cedar Park crash that left two children dead
The identity of the second driver involved in a street race in Cedar Park that led to a fatal car accident that left two children dead and a woman in critical condition on January 10, 2018 has been released.
22-year-old Blake Ayers Kirkpatrick was racing with 18-year-old Hayden Michael Hammer in the westbound lanes of West Whitestone Blvd prior to Hammer crashing into Salman's Nissan, which was turning left onto Walton Way.
The crash happened around 3:45 p.m. on Wednesday, January 10.
Two children, one-year-old Daniel Chaudhary and eight-year-old Elijah Chaudhary, were killed in that accident. Daniel's mother and Elijah's stepmother, 31-year-old Zara Salman, remains in a coma at a local hospital in critical condition.
Hammer has been arrested and charged with two counts of racing on a highway causing death. Both of those charges are second degree felonies. "Mr. Hammer has been cooperating with the investigation and, in an interview with investigators, admitted that he had been drag racing the red or maroon Ford truck prior to the collision," Wukasch said.
Hammer also faces two previous charges out of Taylor County for theft and criminal mischief.
At the time of the crash he was out on bond for the theft charge.


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For more than two years, WND has been at the forefront of reporting a wave of racial violence in the U.S., manifested in what many news outlets are at last acknowledging as "the Knockout Game."
But when the U.S. Justice Department finally decided to act upon on the trend of typically black violence, the Obama administration filed its first hate-crime charges against a white man, Conrad Alvin Barrett of Katy, Texas.
"The charge marks the first time the administration has taken action on a 'Knockout' case after the game became an Internet and media phenomenon," the Washington Times reported. "It chose a case in which the person accused is white, even though most other cases reported in the news have involved black assailants."
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The disparity between the trend and the DOJ's charges has led several voices to conclude there may be more than irony behind the administration's actions.
TRENDING: 'This is an invasion!': Democrat blasted for defending sanctuary policies at Laken Riley briefing
Colin Flaherty, who researched and documented hundreds of cases of racial violence for his 2013 book, "White Girl Bleed A Lot: The Return of Racial Violence to America and How the Media Ignore It," told WND the administration may be using the case to manipulate public opinion on the Knockout Game.
"I have uncovered hundreds of recent examples of black mobs and black teens taking violent actions toward specifically white targets. There can be little doubt there is a racial motivation behind these crimes," Flaherty told WND. "Where have Obama and his DOJ been until now?
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"White people don't have a monopoly on hate," Flaherty continued, "and it smacks of some alternate agenda for this administration to ignore scores of black hate crimes until a white hate crime is found to finally pounce upon."
Flaherty suggested that "agenda" may be to keep the state of racial tensions in the U.S. under wraps.
"For years, there has been an effort to scrub news stories about these crimes of any details about the assailant's race," he explained. "The press has been so determined to avoid any mention of violence among blacks, that it has created a false narrative that it's only 'youths' committing these crimes. I have to wonder if the administration didn't deliberately seize upon this case, knowing the publicity would further the narrative there's no problem with black violence in America, just random violence."
Allen West, the black former congressman from Florida, also questioned whether the charges don't reflect a political agenda on the part of U.S. Attorney General Eric Holder.
"I am glad the U.S. Department of Justice has finally stepped forward, but it begs the question, what took so long?" West asks on his blog. "Does Eric Holder's inaction regarding previous crimes involving blacks attacking whites actually reflect his own hate crime?"
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West continues, "The real question behind the decision to prosecute Mr. Barrett is not his actions, but rather the perception of an even more heinous double standard emanating from the Obama administration's Department of Justice under Eric Holder – sure seems like it's being used as a platform for the progressive socialist version of social justice."
"If the Barrett case were the beginning of a trend of the DOJ equitably prosecuting these brutal and senseless crimes as hate crimes, then that would be all well and good," chimed in Arnold Ahlert at FrontPage Magazine. "However, the notion that the DOJ's newfound interest in prosecuting this particular type of violence is colorblind remains doubtful."
Horace Cooper, constitutional attorney and co-chairman of Project 21, the National Leadership Network of Black Conservatives, told OneNewsNow the DOJ's selective enforcement of hate-crime legislation may be risking the laws' constitutionality.
"The only case that they're able to find is the one example out of a hundred where the victim was black and the perpetrator was white," Cooper said. "That's a very curious outcome."
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Cooper said it appears the Justice Department believes "only whites can engage in hate crime and not blacks."
"If the only way and the only time [a hate-crime law] is ever going to be invoked is to somehow target white Americans, and not target all criminals equally, it becomes constitutionally suspect," says the Project 21 spokesman.
Barrett faces up to 10 years in federal prison for violating the Matthew Shepard and James Byrd Jr. Hate Crimes Prevention Act. Barrett allegedly videotaped himself sucker-punching a 79-year-old black man in a "Knockout Game" just days before Thanksgiving.
According to prosecutors, the attack happened Nov. 24, but it wasn't until 12 days later that authorities connected the attack to the cell-phone video of it.
In the video, according to a federal affidavit filed in the case, Barrett allegedly said: "The plan is to see if I were to hit a black person, would this be nationally televised?"
He then allegedly "hit the man with such force that the man immediately fell to the ground. Barrett then laughed and said 'knockout,' as he ran to his vehicle and fled."
The victim suffered two jaw fractures and was hospitalized for several days, the complaint said.
In the racially charged and sometimes deadly "Knockout Game," one or more assailants, usually black, target a randomly selected white person and, for amusement, try to knock out the unsuspecting victim with a single punch. Nationwide, there have been at least six deaths attributed to the Knockout Game and hundreds of cases have been documented in the book "White Girl Bleed A Lot."
The most recent example of the "Knockout Game" involves a couple taking a late-night stroll at the open-air mall in Charlottesville, Va., when they were viciously beaten.
Jeanne Doucette and her boyfriend Marc Adams were enjoying the Christmas lights downtown when Adams tripped and was immediately pounced upon by three black men.
They beat him severely, breaking his ankle, cracking ribs and knocking out one of his teeth, reports the Charlottesville Weekly.
While Doucette suffered bruising to her head and tearing of the cartilage in her ear, Adams bore the brunt of the men's aggression, sustaining broken bones and a concussion that he said has robbed him of any memory of the incident and its immediate aftermath.
According to the victims, the attackers never uttered any racial epithets and therefore police are not classifying it as a "hate crime."
The assault moved up the street as Adams and Doucette tried to escape their attackers. They screamed at passersby for help. They screamed at their attackers to stop. No one listened. The beating continued.
Then came the laughter, one of the identifying aspects of the"Knockout Game."
"They were laughing, high-fiving, hugging, and then returning to kick him," said Doucette. "There was some kind of camaraderie to it."
Doucette managed to take photos that appear to show the assailants as they kicked and pummeled her boyfriend into unconsciousness.
Nearly two weeks after the attack, the physical wounds are healing, but both Doucette and Adams are troubled by what they see as a lack of response from the Charlottesville Police Department. Doucette said police refused to release her photos to the public.
"I don't understand why they couldn't even have the courtesy to call and say we're not even going to look for them," she told the newspaper.
On Dec. 29, Doucette posted the pictures she'd taken to her Facebook page and said she quickly received several tips that she has passed on to police.
Her frustration at the lack of an investigation is palpable.
"I feel forgotten about," Doucette said. "I feel like I'm not safe."
As WND reported, lawmakers in New York are fighting back against the "Knockout Game," proposing it be made a felony and calling on President Obama and Attorney General Eric Holder to respond.
After years of establishment media silence about the Knockout Game, the New York City attacks pushed the vicious pastime into the national media spotlight. However, the damage from the game was evident years ago. As early as July 2011, the American Thinker featured an article on the Knockout Game.
The media is slowly starting to acknowledge that the attacks are a form of racial violence. A New York City CBS outlet reports:
There have been seven so-called "knockout" or "polar bear" assaults in the Crown Heights and Midwood sections of Brooklyn… The alternate name "polar bearing" comes from the fact that the victims are white.
In the Huffington Post, Al Sharpton wrote that the attacks are racist:
These kids are targeting innocent people, and in many cases specifically targeting Jewish folks. We would not be silent if it were the other way around, and we will not be silent now. This behavior is racist, period.
See a trailer for "White Girl Bleed a Lot":


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'Tis the season for cookies! I love Christmas cookies and one of my favorites is an adaptation of Red's M&M Cookies that I have been making for years. When I first started blogging I was invited to host a cookie party and the recipe was included in the box of goodies for the party and I've been making them ever since. In true Jen fashion, I've been adjusting and tweaking a recipe to fit what I love. I generally always add more vanilla and salt to every recipe, I just love that combo. While I call these Christmas cookies mainly because of the red and green M&Ms, you could most certainly make these cookies any time you'd like. Heck, pick out just the green and do St. Patrick's Day. Go crazy!
Now my goal with chewy sugar cookies is to have more of a sugar cookie with a few M&M, not a Chocolate Chip style M&M cookie. I feel like there is a difference. Our local grocery store has more of a sugar cookie feel with the M&Ms feeling like an afterthought more than anything. Either way, you can't go wrong.
Let's get to the cookies, but before we do that, how cute is this set from Precious Moments? They sent it as a gift last year, and it is adorable!
Don't these look like heaven?
Christmas M&M Cookies
These chewy festive Christmas M&M Cookies are the perfect sweet treat for your holiday cookie swap or a couple for Santa.
- 1 cup (2 sticks) butter
- 2/3 cup brown sugar
- 2/3 cup sugar
- 1 egg
- 2 tsp vanilla extract
- 2 cups flour
- 1 tsp baking soda
- 1 tsp salt
- 1 1/2 cups M&M'S® Brand Milk Chocolate Candies
- Preheat oven to 350°F.
- In a large bowl, cream butter and both sugars until well blended. Add the egg and vanilla extract and mix to combine.
- In a separate bowl, sift the flour, baking soda, and salt together. Slowly incorporate ingredients into the butter mixture and stir to combine.
- Fold in the M&M'S® and chill the dough for 1 hour or overnight.
- Use a cookie scoop and set onto a lightly greased tray, about 2 inches apart.
- Bake for 8-10 minutes for chewier cookies.
Other Cookie Recipes
Check out my other cookie recipes including