Influence of D159687 on OR trial performance.

<p>Effects of D159687 on easy (open bars) versus difficult task (black bars). Dose-dependent improvement in (A) mean percent correct first reach on difficult task performance, with modest improvement on easy trial performance. Individual animal performance plot in easy trials (B) and in difficult trials (C). (D–E) Dose-dependent reduction in the total number of reaches (D) and barrier reaches (E) on difficult taks. (A, D, E) Values are listed as mean ± SEM (n = 8 for vehicle, low and mid-dose groups, n = 7 for high dose group). Asterisks denote significant differences from vehicle treatment (* p<0.05, **p<0.01 and ***p<0.001) following repeat measures one-way ANOVA and Dunnett's post-hoc analysis (n = 7, due to non-completer 7A5D).</p

Influence of D159797 on OR trial performance.

<p>Effects of D159797 on easy (open bars) versus difficult tasks (black bars). Dose-dependent improvement in (A) mean percent correct first reach on difficult task performance. Individual animal performance plot in easy trials (B) and in difficult trials (C). (D–E) Dose-dependent reduction in the total number of reaches (D) and barrier reaches (E) on difficult tasks. (A, D, E) Values are listed as mean ± SEM (n = 8 for vehicle and low dose, and n = 7 for mid and high dose group). Asterisks denote significant differences from vehicle treatment (* p<0.05, **p<0.01 and ***p<0.001) following repeat measures one-way ANOVA and Dunnett's post-hoc analysis (n = 7, due to non-completer 7A5D).</p

Object retrieval task schematic and baseline characterization.

<p>(A) Order of object retrieval task sessions for easy and difficult trials. Drawings illustrate the position of the reward in the boxes. (*) Left position will become right and right position will become left and so on in weekly rotation throughout the study. (#) The <b>bold</b> side of the cube represents the open side of the cube. (**) Trial 17 was for reward purposes and was not included in the data analysis; (B) Box (line indicates median, * indicates mean, box represents upper and lower 25 percentiles) and whisker (maximum to minimum) plots of all animal performance during the 4 training sessions and to vehicle administration during the testing phase on both easy (grey) and difficult (black) trials. Dashed lines indicate targeted performance – performance in easy trials >50% correct first reach, and performance in difficult trials <40% correct first reach. (C) Same data as in (B) but with high performer animal 3939 excluded. Criteria are largely met by exclusion of this animal. The outlier value during the rolipram vehicle trial is due to animal 7A5D.</p

Pharmacokinetic analysis of PDE4D NAMs in female Cynomolgous monkeys.

<p>(A–B) Plasma exposure of D159687 (A), or D159797 (B) in female Cynomolgus monkey plasma following a single intravenous administration at 1.0 mg/kg, and on day 1 and day 7 after repeated daily oral administration at 5.0 mg/kg.</p

Summary of plasma pharmacokinetic parameters of D159687 following single intravenous administration at 1.0/kg, and on day 1 and day 7 after repeated daily oral administration at 5.0 mg/kg.

<p>Data presented as mean ± SD of 3 animals.</p><p>*Calculated from n = 2, as due to patency issues in catheter of one animal following IV dosing, one animal was replaced for po dosing, thus, it was not a cross over design.</p

Summary of plasma pharmacokinetic parameters of D159797 following single intravenous administration at 1.0/kg, and on day 1 and day 7 after repeated daily oral administration at 5.0 mg/kg.

<p>Data presented as mean ± SD of 3 animals.</p

Precision of farmerbased fertility ratings and soil organic carbon for crop production on a Ferralsol

Peer Revie

Blockade by A_2AR antagonists of striatal glutamate release induced by cortical electrical stimulation.

<p>(a) Representative coronal sections of a rat brain, stained with cresyl violet, showing the tracks left by the bipolar stimulation electrode in the orofacial area of the lateral agranular motor cortex (top) and by the microdialysis probe in the lateral striatum (bottom). (b) Effect of systemic administration of the A_2AR antagonists SCH-442416 and KW-6002 (1 mg/kg, i.p., in both cases) on the increase in glutamate extracellular levels in the lateral striatum induced by cortical electrical stimulation. Results are expressed as means ± S.E.M. of percentage of the average of the three values before the stimulation (n = 5–7 per group). Time ‘0’ represents the values of the samples previous to the stimulation. The arrow indicates the time of systemic administration. The train of vertical lines represents the period of cortical stimulation. *: <i>p</i><0.05 compared to value of the last sample before the stimulation (repeated-measures ANOVA followed by Tukey's test).</p

Identification of receptor heteromers in CHO cells by BRET saturation curve.

<p>BRET experiments were performed with CHO cells co-expressing A_2AR-<i>RLuc</i> and A₁R-YFP (A) or A_2AR-<i>RLuc</i> and D₂R-YFP (B). Co-transfections were performed with increasing amounts of plasmid–YFP (0.25 to 4 µg cDNA corresponding to A₁R-YFP and 0.5 to 8 µg corresponding to D₂R-YFP) whereas the A_2AR-<i>RLuc</i> construct was maintained constant (0.5 µg cDNA). Both fluorescence and luminiscence of each sample were measured before every experiment to confirm similar donor expressions (about 100,000 luminescent units) while monitoring the increase acceptor expression (10,000–25,000 fluorescent units). As a negative control, linear BRET was obtained in cells expressing equivalent luminescence and fluorescence amounts corresponding to A_2AR-<i>RLuc</i>, (0.5 µg transfected cDNA) and serotonin 5HT_2B-YFP (0.5 to 8 µg transfected cDNA) receptors. The relative amount of acceptor is given as the ratio between the fluorescence of the acceptor minus the fluorescence value of cells expressing the donor alone (YFP) and the luciferase activity of the donor (Rluc). BRET data are expressed as means ± S.D. of 4–6 different experiments grouped as a function of the amount of BRET acceptor.</p

Validation of the genes predicted to be downregulated in <i>Hdac4</i> knock-out P3 brains.

<p>(A) <i>Nrxn1</i> (neurexin1), <i>Gmbf</i> (glia maturation factor beta), Cdh7 (cadherin 7), <i>Bmpr2</i> (bone morphogenic protein receptor, type II), Stk25 (serine/threonine kinase 25), <i>Atrx</i> (alpha thalassemia/mental retardation syndrome X-linked homolog) and <i>Eif4g1</i> (eukaryotic translation initiation factor 4, gamma 1) were modestly upregulated in <i>Hdac4</i>KO P3 brains. <i>Calml4</i> (calmodulin-like 4) was the only gene that was downregulated (B) The expression level of Hif1α (hypoxia inducible factor 1 alpha subunit), <i>Crebbp</i> (CREB-binding protein), Syt1 (synaptotagemin 1) <i>Garln1</i> (GTPase activating RANGAP domain like 1 protein), <i>Rapgef6</i> (Rap guanine nucleotide exchange factor GEF6), Rock1 (Rho-associated coiled-coil containing protein kinase 1), <i>Nedd4</i> (NEDD4 binding protein), <i>Jmjd4</i> (jumonji containing protein 4), <i>Casc4</i> (cancer susceptibility candidate 4), <i>Pnmal</i> (PNMA-like 2), <i>Rlf</i> (rearranged L-myc fusion sequence) and Scd2 (stearoyl-Coenzyme A desaturase 2) did not change between <i>Hdac4</i>KO and WT P3 brains. All mRNA expression levels were assessed by Taqman qPCR and presented as a relative expression ratio to the geometric mean of three housekeeping genes <i>Atp5b</i>, <i>Canx</i>, <i>Rpl13α</i>. Error bars are S.E.M (n>7). *<i>p</i><0.05, **<i>p</i><0.01.</p