Brief Summary of Minutes of Annual Meeting

Accomplishments and Impacts The overall objectives of this Multistate Regional Project are:
Objective #1: Evaluate the effect of different n-3 fatty acids, both form (source) and amount, on tissue functions and correlate these effects with changes in putative biomarkers relevant to health promotion and disease prevention.
Objective #2: Experimental diets used in animal studies will examine dietary levels of n-3 PUFA that are achievable in human diets, based on human equivalent amounts (allometric scaling) in rodent models.
Objective #3: Develop effective means for translating research knowledge about n-3 PUFA into consumer food choices.

The outcomes and impacts of this Multistate Regional Project are:
" Improved health of the US population.
" Decreased incidence of age-related and/or degenerative diseases.
" Education of clients by nutrition professional about health benefits of n-3 PUFA, both forms and amounts.
" Oil processors will be introduced to the concerns health professionals have about meeting n-3 PUFA guidelines.
" Stimulating the development of alternative sources of agriculturally-based LCn-3 PUFA.
" Stimulating the development and marketing of new and creative agriculturally-based food products containing LCn-3 PUFA.
" Enhanced awareness by the medical community of the benefits n-3 PUFA consumption.

Milestones are:
" Collection of data to be used as preliminary data for competitive grant submissions.
" Submission of competitive grant applications to USADA and NIH.
" Objective #3 is addressed in three consecutively dependent stages: (1) Development of Nutrition Education Intervention; (2) Implementation of the Intervention; and (3) Evaluation of Outcomes.

Accomplishments and Impacts

Experiments in animals have addressed the importance of preformed long-chain n-3 polyunsaturated fatty acids (LC n-3 PUFA) in indices of premature delivery and cardiovascular disease.


Experiments have shown that substitution of long-chain n-3 polyunsaturated fatty acids (LC n-3 PUFA), such as eicosapentaenoic acid (EPA), docosahexaenoic acid (DHA) and stearidonic acid (SDA), for linolenic acid (LnA) improves markers of premature delivery, ovulation and cardiovascular disease. For indices of premature delivery both DHA and SDA provided at 0.7 % of calories, the current US intake of total n-3 PUFA, were effective in reducing placental prostaglandin production. DHA provided at 0.7 % of calories also reduced placental collagenase and matrix metalloproteinase production. Both prostaglandin production and the amounts of collagenase and matrix metalloproteinases are important determinants of parturition, premature delivery and infant birth weight. These results show that substituting a LC n-3 PUFA was more effective than increasing LnA intake. Thus, a switch to foods providing these LC n-3 PUFA, without increasing the intake of n-3 PUFA is effective in improving markers of premature delivery and gestational duration both of which are major health problems for infants.


Heart disease remains the major cause of mortality in the US and inflammation plays a major role in its development. Interleukin-6 (IL-6), a proinflammatory cytokine is secreted by a variety of cells and plays a role in the production of C-reactive protein which increases cardiovascular disease risk by increasing uptake of low density lipoprotein cholesterol. Anti-inflammatory n-3 fatty acids such as EPA (20:5, n-3) and SDA (18:4, n-3) inhibited lipopolysaccharide (LPS)-induced increases in IL-6 secretion in murine 3T3-L1 preadipocytes. In a cell culture experiment, data showed that preadipocytes secreted significantly higher levels of IL-6, and treatment of preadipocytes with both EPA and SDA decreases LPS-induced IL-6 secretion significantly. As a result, this demonstrates that one of the ways in which n-3 fatty acids can reduce cardiovascular disease risk may be via down regulating IL-6 in the preadipocytes. These results suggest that LC n-3 PUFA are effective in preventing some of the inflammatory processes of cardiovascular disease.
These results address Objectives 1 and 2 and the first milestone of our original multistate proposal. In particular, the form of n-3 PUFA (EPA, DHA, SDA) has been identified as more important than providing more linolenic acid. This has important implications for human health in that consumers believe that all forms of n-3 PUFA have health benefits. The data from this project indicate that preformed LC n-3 PUFA are much more important in providing health benefits. Even increasing LnA intakes 3-fold to 2.0 % of calories does not afford the same benefits as simply substituting preformed LC n-3 PUFA for the current 0.7 % of calories provided by LnA.


Despite the benefits of increased LC n-3 PUFA intakes, there is concern that infectious disease resistance is impaired by n-3 PUFA. Disease resistance using the murine listeriosis model has been examined. Listeriosis is an infectious disease of humans and animals that is caused by the intracellular, gram-positive bacteria, Listeria monocytogenes. This pathogen has been used by many investigators as a model organism to define the various factors and cells that are involved in the host response to intracellular pathogens. Cells of the innate immune system (i.e. macrophages, natural killer cells, neutrophils, dendritic cells) play a critical role in controlling bacterial growth during the initial stages of infection. It is this stage of host defense that seems most seriously impaired by LC n-3 PUFA, such as those provided by fish oil. The consequences are best illustrated by the reduced bacterial clearance we have observed in the liver and spleen 3 to 4 days post-infection. Typically at this time, we have found 10 to 100-fold more bacteria in the liver and spleen of mice fed diets rich in LCn-3 PUFA compared with mice fed diets rich in n-6 PUFA or low in total PUFA (i.e., soybean oil and lard, respectively). It has been shown that changes in bacterial clearance correlate with lower survival rates for mice consuming a high LCn-3 PUFA diet. We have some evidence that both LCn-3 PUFA found in fish oils eicosapentaenoic acid (EPA, 20:5n-3) and docosahexaenoic acid (DHA, 22:6n-3) are equally effective at modulating host infectious disease resistance to this pathogen. However, we have not examined other n-3 PUFA, nor have we addressed the dose-response relationship between n-3 PUFA and host resistance. To date we have observed a significant adverse effect of LCn-3 PUFA at a level of intake equivalent to ~2 energy%. This is about 10-fold higher than the current average intake of LCn-3 PUFA for people in the U.S., however, it is not far from the levels that are currently being recommended by some health professionals (i.e., 0.65% of calories). These studies address objectives 1 and 2 and the first milestone of our original proposal.
In order to disseminate the results from animal experiments members of the NC 1167 committee have begun to address the dietary behavior patterns that influence omega-3 fatty acid intakes, and to design effective omega-3 nutrition education interventions. This year we have developed and pilot tested an n-3 fatty acid food frequency.


We have designed and evaluated an omega-3 fatty acid nutrition education intervention for heart patients and have shown that participants increased their n-3 fatty acid intakes after receiving the education. We have also assessed the impact of geographic region on omega-3 fatty acid intakes by surveying the intakes of elderly men in two geographic regions (a coastal and an internal region) of Saudi Arabia. Residents of the coastal region have significantly higher intakes than those in the internal regions of the country.


IMPACT: Increasing n-3 fatty acid intakes will have a favorable impact on reducing risk of several chronic diseases such as cardiovascular disease, arthritis, and cancer and in the long term will reduce health care costs and increase quality of life for the elderly. Accurate assessment of n-3 fatty acid intakes requires effective nutrition education interventions for increasing n-3 fatty acid intakes. Understanding of factors that influence n-3 fatty acid intakes, such as geographic location, provide components and information needed by nutrition educators to increase n-3 fatty acid intakes in their clientele.
During this past year, NC-1167 partners from Kansas, Colorado, New Jersey and Nebraska have worked as a team to develop a questionnaire for assessing omega-3 knowledge and practices of dietitians. For the formative phase of development, 10 dietitians from each state were called and ask to respond to open-ended questions. Responses to each open-ended question were grouped and from this a detailed questionnaire was developed. The detailed questionnaire was then adapted for completion using the web and this web survey was piloted with two dietitians from each state and further refinements have been made. Our plan at this time is to conduct the survey early in 2004 and to obtain responses using both telephone and web-based surveys. Information gathered from these surveys will provide a profile of baseline knowledge and practices of a random sample of dietitians in these four states.


IMPACT: Consumers look to practicing dietitians as a source of accurate and unbiased information on nutrition. By assessing baseline knowledge and practices of dietitians we can understand their n-3 nutrition education needs and can design educational resources to meet these needs.
These studies addressing dietary behavior patterns that influence omega-3 fatty acid intakes, and the design of effective omega-3 nutrition education interventions address Objective 3 and the third milestone of our original proposal.

Turek, J.J., Watkins, B.A., Schoenlein, I.A., Allen, K.G.D., Hayek, M.G. and Aldrich, C.G. Oxidized lipids depress canine growth, immune function, and bone formation. J. Nutr. Biochem. 14: 24-31, 2003.


Yu, L., Perret, J., Parker,T. and Allen, K.G.D. Enzymatic modification to improve the water-absorbing and gelling properties of psyllium. Food Chem. 82: 243-248, 2003.


Watkins, B.A., Li, Y., Romsos, D.R., Hoffman, W.E., Allen, K.G.D. and Seifert, M.F. CLA and Bone Modeling in Rats. In: Advances in Conjugated Linoleic Acid Research, Volume 2, pp 218-250 (Sebedio, J-L., Christie, W.W. and Adlof, R. Eds.) American Oil Chemists Society (AOCS) Press, Champaign IL, 2003.


Allen, K.G.D., Perez, M.A., Hansen, R.A. and Harris, M.A. N-3 fatty acids alter eicosanoid and matrix metalloproteinase mediators of parturition in rats. Presented at Experimental Biology 2003 Meeting, San Diego, CA, April 11-15, 2003. FASEB J. 17, A1118 (Abstract # 695.1), 2003.

Hansen, R.A. Short term effects of dietary fish oil on matrix metalloproteinases in dogs with anterior cruciate ligament disease. PhD Dissertation, Colorado State University, Fort Collins, CO, Summer 2003.

Seal, S.J. Circulating PGE2 and PGF2? metabolite and long chain polyunsaturated fatty acid levels in full term pregnancy with docosahexaenoic acid supplementation. Masters Thesis, Colorado State University, Fort Collins, CO, Spring 2003.

Hansen, R.A., Allen, K.G.D., Pluhar, E.G., Ogilvie, G.K., Fettman, M.J. and Harris, M.A. N-3 fatty acids decrease inflammatory mediators in arthritic dogs. Presented at Experimental Biology 2003 Meeting, San Diego, CA, April 11-15, 2003. FASEB J. 17, A330 (Abstract # 200.3), 2003.


Allen, K.G.D. A role for dietary n-3 fatty acids in gestational length. Presented at The Institute of Food Technologists (IFT) Annual Meeting and Food Expo, July 12-16, 2003, Chicago IL, as part of Session 82: Symposium on Dietary n-6/n-3 Fatty Acid Balance. Abstract 82-3, page 207 of 2003 IFT Annual Meeting Technical Program Abstracts.


Lee, J-Y., Lewis, N.M., Scheideler, S.E., Carr, T.P. 2003. Consumption of omega-3 fatty acid enriched eggs and serum lipids in humans. Journal of Nutraceuticals, Functional and Medical Foods. 4(1):3-13.


Lewis, N., Plugge, S., Eskridge, K., Wander, R.C. 2003. Serum ALA increases with inclusion of one omega-3 enriched egg in the diet. The FASEB J. 17:A336.


Al-Numair, Khalid and Lewis, Nancy M. 2003. Variation in source and consumption of omega-3 fatty acids in two different population groups in Saudi Arabia. 5th International Food Data Conference and 27th National Nutrient Databank Conference, Washington, D.C., Proceedings, p. 85.


Heidal, Kimberly. 2003. Evaluating the effects of a nutrition education program on the consumption of omega-3 fatty acids for heart patients. Ph.D. Dissertation, University of Nebraska, Lincoln, NE.


Lohse B, Shafer K. Problem-based learning is effective for extension education about omega-3 fats. Society for Nutrition Education Annual Conference Proceedings 36(1):33;2003.


Lohse B, Nitzke S, Ney D. Introducing a problem-based unit into a lifespan nutrition class using a randomized design produces equivocal outcomes. J Amer Diet Assoc. 103(8):1020-1025;2003.


Lohse Knous B, Nutritional Issues for Pharmacists: Dietary Supplements [Continuing Professional Education] 2003;29(4):1-23. Lawrence, KS:Pharmat, Inc.


Weigh to Diet. Lesson (MF2595) and Leader?s Guide (MF2596). August 2003.
http://www.oznet.ksu.edu/library/fntr2/MF2596.pdf
http://www.oznet.ksu.edu/library/fntr2/MF2595.pdf