1.What major problem or issue is being resolved and how are you resolving it (summarize project aims and objectives)? How serious is the problem? What does it matter?
The amount and type of dietary fatty acids plays an important role in health maintenance and disease prevention. Different fatty acids and their metabolites regulate signal transduction, gene expression, cell growth, differentiation, and death. Excessive intake of omega-6 fatty acids found in most seed oils has the potential to increase inflammatory diseases, while omega-3 fatty acids found in marine and some seed oils (flax, perilla, and walnuts) have strong anti-inflammatory effects; the ratio between the dietary omega-6 and omega-3 fatty acids influences the development and progression of several human chronic diseases including cardiovascular disease (CVD), diabetes, arthritis, lupus, inflammatory bowel disease, and many others. CVD alone is responsible for more than one-half of all the deaths in USA and many European countries. Overall goal of our studies is to determine the effects of dietary fatty acids on immune and inflammatory responses and on risk factors of chronic diseases. Specific objectives are: 1. Determine effects of DHA supplementation on risk factors for CVD in hypertriglyceridemic men. 2. Determine effects of omega-6 and omega-3 fatty acids on proliferation and differentiation of granulocytic precursor cells, and elucidate the mechanisms involved. 3. Determine whether different dietary fatty acids modulate the activation of Toll-like receptors, downstream signaling pathways, target gene expression and consequent cellular responses. Specific aim 1 will involve DHA supplementation to hypertriglyceridemic men, and monitor its effects on a number of conventional and novel markers for CVD, markers for diabetes, and gene expression. Specific aims 2 & 3 will involve feeding diets with different fatty acid compositions to animal models, and adding fatty acids to cultured cells. Results obtained will provide the information needed to determine dietary requirements for long chain PUFA, specific dietary fatty acid based therapies for the prevention and management of inflammatory conditions. This is expected to improve quality of life and may open new avenues for agricultural products. Understanding the mechanisms by which different dietary fatty acids improve health or cause diseases, may identify the metabolic sites that may be regulated by drug therapies as well.

2.List the milestones (indicators of progress) from your Project Plan.
Specific Aim 1:

Year 1: Subject recruitment, fatty acid supplementation, and sample collection. Year 2: Continue with that listed for year 1 and start sample and data analysis. Year 3-5: Sample and data analysis, and preparation of manuscripts.

Specific Aim 2:

Year 3: Conduct experiments with promyelocytic cell lines. Year 4: Analyze data from experiments with cell lines and prepare manuscripts, conduct experiments with mice. Year 5: Analyze samples and data from the mice study, and prepare manuscripts.

Specific Aim 3:

Year 1-3: Determine relative potency of n-3 PUFAs as compared with n-6 PUFAs in inhibiting the activation of toll like receptors (TLR) and if different TLRs are differently modulated by fatty acids.

Year 1-5: Determine whether TLR4 induced activation of downstream signaling pathways and target gene expressions are also altered by fatty acids. (This is a tentative time schedule.

3.Milestones:
Specific aims 1 and 2: In FY 2005, we plan to complete sample collection from the human DHA supplementation study, start experiments with promyelocytic cell lines. In FY 2006, we will continue sample and data analysis from the human study, and cell lines. In FY 2007, we will continue sample and data analysis from the DHA study and those experiments with cell lines and mouse model, and prepare manuscripts. Results obtained from the DHA study may provide new nutritional information that can be used to manage hypertriglyceridemia and other risk factors for CVD; results from studies dealing the effects of fatty acids on granulocyte numbers and activity may provide information to manage neutropenia and neutrophelia.

Specific aim 3: We will continue to study which downstream signaling pathways derived from TLR activation are modulated by fatty acids, and in turn which target genes are differentially modulated. Identification of the differentially regulated genes and consequent cellular responses will eventually lead us to understand why types of dietary fatty acids differentially affect the risk of development of chronic diseases.

4.What were the most significant accomplishments this past year?
A. The benefits and/or risks of individual isomers of conjugated linoleic acid (CLA) to human health are not known. Research conducted by Dr. Kelley at WHNRC evaluated two purified isomers of CLA (c9, t11- and t10, c12-CLA) to determine their effects on lipid metabolism. Both isomers altered the fatty acid composition of liver lipids, but only t10, c12-CLA caused a five fold increase in the amount of liver fat. These results show that CLA isomers differ in their effects on lipid metabolism and the t10, c12-CLA can be potentially very harmful. B. None.

C. None

D. None

5.Describe the major accomplishments over the life of the project, including their predicted or actual impact.
5. This is a new CRIS and the report is for its first year; hence accomplishments listed under question 4 apply here. Our previous CRIS also dealt with the health effects of dietary fatty acids and the accomplishments listed are for the expired CRIS. Research conducted under the expired CRIS showed that increasing total fat intake inhibited several aspects of human immune status. If total fat intake was held constant, increasing the intake of omega-3 fatty acids reduced a number of markers for inflammation (numbers of circulating granulocytes, inflammatory cytokines and eicosanoids), while omega-6 fatty acids had the opposite effects. Omega-3 fatty acids are now used in the management of several inflammatory diseases including, CVD, arthritis, asthma, and adult respiratory distress syndrome or ARDS. We have also shown that the ratio between saturated and unsaturated fatty acids was more critical than the amount of dietary fat in determining serum cholesterol levels. Our studies conducted with CLA supplementation showed no benefit, but several potential risks to humans. Results from our studies have been used by health organizations for making recommendations regarding fatty acid intake.

6.What science and/or technologies have been transferred and to whom? When is the science and/or technology likely to become available to the end-user (industry, farmer, other scientists)? What are the constraints, if known, to the adoption and durability of the technology products?
None.

Review Publications
Journal Biological Chemustry, Vol.278, No.39, Issue of September 26, pp. 37041-37051, 2003