lesioned hemisphere of rat brain on day 7 after tMCAO. Two groups (10 rats per group) were set as control, sham-grafted rats received equivalent volume (6 Al) of vehicle. The behavioral tests were carried out while pathologic examination was processed within 3 months after transplantation. The results revealed that All NSC transplantation in the three doses can improve the neurological function of tMCAO rats. There were significant difference between the group I and group II; however, no significant difference was observed between group I and group III. Conclusions NSCs can be isolated, cultured and passaged in vitro. Transplanted NSCs can survive in the host brain and differentiate into neurons and astrocytes and can promote the functional recovery after tMCAO. The treatment effect is concerned with administrated NSCs dose. In this study, the amount in the middle range of NSCs was considered as an optimal dose to benefit effectively the function recovery of tMCAO rat. doi:10.1016/j.expneurol.2006.02.103 Dietary supplementation with blueberry extract attenuates the age-associated increase in microglial activation L.M. Willis 1 , P.C. Bickford 3 , I. Stromberg 4 , A.C. Granholm 1,2 1 Department of Neurosciences, Medical University of South Carolina, Charleston, USA 2 Center on Aging, Medical University of South Carolina, Charleston, USA 3 Department of Neurosurgery, University of South Florida, Tampa, USA 4 Department of Integrative Medical Biology, Umea University, Umea, Sweden Dietary supplementation with foods high in antioxidants, such as blueberries, has been shown to have dramatic effects in the central nervous system. Although blueberries have tradi- tionally been recognized for their potent antioxidant activity, recent studies indicate that dietary supplementation with this fruit may also affect inflammatory processes in the brain. Central nervous system inflammation is elevated in a number of neurodegenerative disorders and may also be detrimental to neuronal functioning during normal aging. The present study explores the age-related increase in inflammation and the potential of dietary supplementation with blueberry extract to attenuate this inflammation. Young (6 months) and middle aged (19 months) rats were given a diet of either feed supplemented with 2% blueberry extract or a control feed and were maintained on the respective diets for 12 weeks. Inflammatory status was assessed in the form of activated microglia. Microglia that have become activated undergo morphological changes and exhibit expression of the MHC class II molecule OX6. OX6 expression by these cells is indicative of an activated state, in which microglia release toxic cytokines, damaging nearby cells. Preliminary results indicate a robust increase in OX6 positive microglia in the hippocampus of aged versus young animals. This increase was dramatically attenuated in aged animals receiving a diet supplemented with 2% blueberry extract. Other markers of inflammation such as COX-2, IL-6 and TNF-a are currently being examined. Considering that inflammation is associated with neurodegen- erative diseases, brain injury and age-associated cognitive decline, the anti-inflammatory potency of blueberries has profound implications for neuroprotective therapeutics. (Sup- ported by NIH grants AG04418 and AT003023). doi:10.1016/j.expneurol.2006.02.104 An intermediate pathway of cell death involving cathepsin D during tumor necrosis factor-A-induced apoptosis T.A. Womble, D.C. Lee, I.N. Jackson, C.A. Mason, B.A. Smith, N. Anderson, D.E. Palm Florida A&M University College of Pharmacy and Pharmaceutical Sciences, Tallahassee, FL, USA Background: Tumor necrosis factor-alpha (TNF-a) is a pleio- tropic cytokine that is rapidly upregulated in the brain following injury. These elevated levels of TNF-a have been detected in ischemic brain cells in experimental animal models, and are believed to play an important role in the propagation of cell death following a stroke. However, the pathway leading to this cellular demise has been directly attributed to the classical pathway of caspase activation. Following the protection provided by inhibition of the executioner caspase (caspase 3), the continual presence of cell death leads us to investigate the role of an intermediate pathway regulated by lysosomal proteases in N1E- 115 murine neuroblastoma cells following TNF-a-induced cell death. Purpose: Here we show the role of the lysosomal proteases cathepsin B and D in an intermediate pathway triggered by TNF-a. Methods: Characterization of the type of cell death was determined by the use of terminal deoxy-nucleotidyl transfer- ase-mediated dUTP nick-end labeling (TUNEL). Immunocy- tochemistry was utilized to localize cathepsin D following TNF-a treatment. Variations in cathepsin D expression were observed using Western blot analysis. All experiments were carried out using N1E-115 murine neuroblastoma cells treated with TNF-a (1, 25, 50 and 100 ng/ml) for 3, 6, 12, 24, and 48 h. We also quantitatively assessed the cytotoxic effects of N1E-115 murine neuroblastoma cells pretreated with various caspase inhibitors or cathepsin D inhibitor prior to treatment with TNF-a by measuring the release of lactate dehydrogenase (LDH) from damaged cells into the extracellular fluid. Results: There was no significant difference in the 3 and 6 h time point but a time-dependent increase in cell death was observed during 12, 24 and 48 h exposure. We also showed a dose-dependent increase in cathepsin D expression to TNF-a as well as cathepsin D immunoreactivity to TNF-a. Pretreat- ment with caspase inhibitors ACDEVD-cho (caspase 3 inhibitor) and BOC-fmk (pan-caspase inhibitor) revealed partial protection following TNF-a-induced death. Interesting- ly, pretreatment with FMK-007 (caspase 8 inhibitor) prior to TNF-a-induced cell death resulted in total protection. Addi- ABSTRACTS / Experimental Neurology 198 (2006) 558 – 597 594