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United States Department of Agriculture

Agricultural Research Service

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Location: Boston, Massachusetts

2009 Annual Report

1a. Objectives (from AD-416)
1: Determine the effects of SB supplementation on behavioral aging using paradigms sensitive to cognitive (short and long-term memories) behaviors. 2: Determine the effects of SB supplementation on calcium dependent and neuronal signaling (2) and neurogenesis (2b) correlate these with alterations in behavioral parameters determined in SA 1. 3: Determine whether the efficacies of these supplementations in the behavioral assays are associated with enhanced resistance to oxidative stress or inflammation. 4: Establish the effects of nutritional modulation (e.g., berry fruit) on behavioral (e.g., cognition, gait, force and balance) and neuronal deficits in aging to assess the mechanisms involved and the most effective dietary supplements in animal and human models.

1b. Approach (from AD-416)
Aim 1. The effects of SB dietary supplementation (2% SB extract in the diet for 8 weeks. After 8 weeks performance will be examined in the rats using age-sensitive cognitive behaviors that are selective for reference and working memories. The latter behaviors will be assessed using the Morris water maze and the radial arm water maze, as well as novelty tests. All of these tests have been validated as being age sensitive (e.g. demonstrated a significant decline as a function of age). Aim 2. The effects of SB dietary supplementation on neurogenesis and differentiation will be examined using immunocytochemistry, bromodeoxyuridine (BrDU) incorporation in hippocampus and olfactory bulb obtained from the supplemented behaviorally-assessed animals Calcium signaling will be assessed by examining Ca45 clearance in tissues (as above) taken from the brain. Aim 3. For the basal assessments we will use immunohistochemistry to assess various markers of oxidative stress (e.g., HO-1) and inflammation (e.g., cytokines) as well as immunoblotting to detect HO-1 and bcl 2 expression. The responses of the tissue (muscarinic receptor sensitivity, HSP-70 activation) to oxidant (hydrogen peroxide, 10 uM) or inflammatory (LPS) stressors will be assessed by exposing cross-cut slices of the various brain regions obtained from the supplemented animals.

3. Progress Report
Previously, we have shown that whole, crude berry extracts are able to reverse several parameters of brain aging, as well as age-related motor and cognitive deficits when fed to rats from 19-21 months of age. These effects may be the result of direct effects on brain signaling or indirect effects through antioxidant and anti-inflammatory properties of the polyphenols. This year we extended these studies by attempting to determine the mechanisms of the beneficial effects of the strawberries. In this respect we showed that deficits in calcium function induced by the oxidant stressors dopamine or amyloid beta (AB) in cells are blocked by blueberry (BB) and strawberry (SB) extracts and that BB or SB extract pretreatments can reduce both stress- and inflammatory- induced neuronal dysfunction. This is important since if the neuronal cell’s ability to clear calcium is antagonized by oxidative or inflammatory stressors, the cell may lose viability and show declines in function. The finding that BB or SB pre-treatments can mitigate these effects has important implications for preventing declines in neuronal function via nutrition. It appears that the mechanisms involved in these beneficial effects may involve declines in the signals that are enhanced by the stressors. Thus a subsequent study in cells indicated that at least part of BB-induced reductions in stress mediated signaling might involve decreases in nitric oxide which is a powerful stressor. Experiments were carried out with BB and SB to determine if similar findings could be seen in co-cultures of different but related cells sensitive to oxidative stress and inflammation. Results indicated that in the cells the effectiveness of the various treatments are dependent upon the stressor that we used such as dopamine, suggesting that it is better to eat a variety of berries. We showed for the first time that mechanisms other than the inactivation of free radicals (molecules producing oxidative stress) are involved in the beneficial effects of the berry fruits. These findings are important in both industry and health areas, since they point once again to the importance of including berries in the diet for promoting healthy aging.

4. Accomplishments

Last Modified: 10/18/2017
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