Therapeutic protein inhibition of spontaneous calcium oscillations mediated by either NMDA or Watch Live Sexcam AMPA will be assessed. Addition of therapeutic protein will inhibit AMPA receptor mediated oscillations. FLIPR signal was read for an additional 200 seconds (1 reading/second) after the addition of oligomer. Baseline signal (measurement of intracellular calcium) was read for 100 seconds (1 reading/second) before the addition of anti-sense oligomers. A second 5 minute post addition plate read (300 one second points) on the FLIPR was conducted to allow for additional data capture. Raw data from the 5 minute read was exported and, using Excel, spike amplitude and frequency was calculated. Data presented in FIG. 1 show that, addition of the AMPA receptor antagonist, 6-Cyano-7-nitroquinoxaline-2,3-dione (CNQX; 3 .mu.M), reduced calcium oscillations by 20% representing the total AMPA response in the assay (FIG. 1AMPA labeled bar shown). Addition of small molecule will inhibit AMPA receptor mediated oscillations. Small molecule inhibition of spontaneous calcium oscillations mediated by either NMDA or AMPA will be assessed.



Antisense oligomer inhibition of spontaneous calcium oscillations mediated by either NMDA or AMPA was assessed in the presence or absence of 1 mM MgCl.sub.2 (representing 100% control in each case; FIG. 2). Addition of 25 .mu.M antisense oligomers (ASO) inhibited AMPA receptor but not NMDA receptor mediated oscillations (FIG. 2). ASOs, and other oligomers that behaved similarly, were shown to negatively impact central nervous system (CNS) network activity in vivo and electrophysiologic spontaneous neuronal activity in vitro (data not shown). Calcium oscillations were reduced further, by about 80%, when (NMDA) receptor function was blocked by 1 mM MgCl.sub.2 (FIG. 1 NMDA labeled bar shown). The effect of small molecules on calcium oscillations will be measured using substantially the same method provided in Example 2. To measure primary cortical neuron spontaneous calcium oscillation, rat primary cortical neurons will be prepared. The effect of therapeutic proteins, such as antibodies or antigen-binding fragments thereof, fusion proteins, cytokines, cell surface receptors, hormones or growth factors, on calcium oscillations will be measured using substantially the same method provided in Example 2. To measure primary cortical neuron spontaneous calcium oscillation, rat primary cortical neurons will be prepared.



To measure primary cortical neuron spontaneous calcium oscillation, rat primary cortical neurons were prepared from Sprague-Dawley rat embryos (E19). Tau antisense oligonucleotide impact on spontaneous calcium oscillations in primary neurons is shown in FIG. 4. The oligomers in neuronal cells showing AMPA-mediated oscillations that are equal to or greater than 75% of the calcium oscillations in the untreated control cells were selected for further analysis. Tau antisense oligonucleotide impact on spontaneous calcium oscillations in primary neurons is summarized in FIG. 4. See Murphy et al., J. Neurosci. 1987) Gene Transfer Vectors For Mammalian Cells, (Cold Spring Harbor Laboratory); Wu et al., eds., Methods In Enzymology, Vols. Miller, watch live sexcam A.D., "Human gene therapy comes of age." Nature (1992); 357: 455-460. cited by applicant . The human mind is an incredible thing. As discussed in Example 2, the effect of therapeutic proteins on primary neuronal spontaneous calcium oscillations will be measured under two conditions, in the presence and Watchlivesexcam.Com absence of 1 mM MgCl.sub.2 as described previously (Murphy et.



Therapeutic proteins that reduce calcium oscillations to levels below 70% of control will be expected to negatively impact CNS network activity in vivo and electrophysiologic spontaneous neuronal activity in vitro. Small molecules that reduce calcium oscillations to levels below 70% of control will be expected to negatively impact CNS network activity in vivo and electrophysiologic spontaneous neuronal activity in vitro. Calcium oscillation reduction in the neuronal cells was measured for the oligomers of the invention and compared with that of the control cells (i.e., the calcium oscillations in the neuronal cells that are not treated with the oligomers). Oligomers were added with a 384 well head in the FLIPR in 20 .mu.l of assay buffer at 75 .mu.M for a final concentration of 25 .mu.M. After 1 hr 20 .mu.l of room temperature assay buffer was added and the cells were allowed to equilibrate to room temperature for 10 additional minutes and placed in the fluorescent imaging plate reader (FLIPR). Cells will be incubated and allowed to equilibrate to room temperature to measure the fluorescent intensity. We're always working towards adding more features that will keep your love for porno alive and well.



Other features include email, web cam and blogs. As discussed in Example 2, the effect of small molecules on primary neuronal spontaneous calcium oscillations will be measured under two conditions, in the presence and absence of 1 mM MgCl2 as described previously (Murphy et. Effect of oligomers on primary neuronal spontaneous calcium oscillations was measured under two conditions, in the presence and absence of 1 mM MgCl2 as described previously (Murphy et. Sequence score is a mathematical calculation determined for all oligomers and is based on the percent of G and C nucleotides, or analogs thereof, within a given oligomer sequence. For treated wells, a scoring system was developed where a score of 1 was given for each 1 second read where signal increase greater than 50% of the average control value (calculated above). A score of 0 was given for each 1 second read that increase less than 50% of average control value.