2007 Annual Report 1a.Objectives (from AD-416) Aim 1: Continue to determine the effects of BB supplementation on behavioral aging using paradigms sensitive to: motor (probes sensitive to balance, strength, coordination) and cognitive (short- and long-term memories) behaviors. Aim 2: Determine the effects of BB supplementation on signaling and neurogenesis and correlate these with alterations in behavioral parameters determined in SA1. Aim 3: Determine the mechanisms involved in the anti-inflammatory/antioxidant activity of BB in cells. 1b.Approach (from AD-416) After 8 weeks supplementation of 2% (of the blueberry diet), performance will be examined in the rats using age-sensitive motor (e.g., accelerating rotarod) and cognitive (Morris water maze) behaviors. The effects of BB dietary supplementation on neurogenesis proliferation and differentiation will be examined using immunocytochemistry, i.e., bromodeoxyuridine (BrDU) incorporation in hippocampus obtained from the supplemented behaviorally-assessed animals. We will also assess the effects of BB on kainic acid and lipopolysaccharide (LPS) treated BV-2 microglial cells and COS-7 cells will be examined by assessing cell death and calcium clearance. Recently, we have found that several markers of inflammation, including the cytokine, Il-1 beta, the transcription factor, NfkappaB, are both suppressed when the BV-2 cells are pre-treated with BB before being exposed to LPS or kainic acid. We will continue these experiments under the present proposal to determine if the antioxidant/anti-inflammatory properties of BB involve reductions in inflammatory/oxidant signals. We will use imaging, Western blot, and gene array with real-time PCR analyses for these experiments. 3.Progress Report This report serves to document research conducted under a Trust Fund Cooperative Agreement between ARS and the Wild Blueberry Association of North America. Last year we assessed three different BB-derived diets, all equated on phenolic levels, to determine whether the effects observed with the whole, crude BB extract are due to polyphenolics or if the other compounds were contributing to the age-related improvements in behavior. We choose to examine fresh plum juice in this phase of the study for its effectiveness in reversing age-related behavioral and brain effects, and their potential mechanism(s) of action. Nineteen month old F344 rats (15/group x 2 groups, for a total of 30 rats) were given either water or plum juice (18.5 Brix, provided by Sunsweet) as their sole source of liquid before behavioral testing (as described above). The water or juice was provided to the rats in small glass jars secured to their cages to prevent spillage. The experimental products in the jars were changed daily and the jars were changed twice a week. The plum juice rats performed significantly better on the cognitive tests, but not the motor tests, than did control rats. When examining cognitive performance, we performed separate t-tests between the two trial latencies for each group, in order to see if the different juice groups significantly improved their performance from Trial 1 to Trial 2, showing improved working memory. We found that the plum juice group showed significant (p < 0.05) differences between Trial 1 and Trial 2, i.e., Trial 2 latencies and distances to the platform were significantly less than Trial 1, showing that these rats demonstrated one-trial learning, even with the 10 min retention interval. This one-trial learning was not found in the water group. Weights showed no overall group differences.therefore, although there were no effects on motor function the plum juice had a significant effect on cognitive performance. Extrapolating from the effects previously seen with blueberries, these findings suggest that perhaps the polyphenolics in the plum juice may be having some profound effects on neuronal communication, that is-- "getting old neurons to talk to one another again". If this is the case then we would hope to explore this further in additional experiments in the next Phase of this research. Additional details of this research can be found in the report for the parent CRIS 1950-51000-063-00D Nutritional Modulation of Brain Aging and Cognitive Decline.