Brief Summary of Minutes of Annual Meeting

The meeting was called to order by the chair, Kevin Fritsche at 8:30 a.m., October 6, 2005 at the Fort Collins Hilton, Fort Collins, CO. Kate Claycombe served as a secretary for this meeting. Introductions & Opening Remarks: The meeting opened with an around the introduction of participating members and a welcome by April Mason, Dean of College of Applied Human Sciences at Colorado State University. Dean Mason, with her background in Nutrition, commented on importance of our group's potentially contribution or impact on American public's dietary n-3 PUFA intake modification. Susan Welsh was not able to attend the meeting due to other on-going USDA task (a self study report). Susan; however, sent a report via e-mail to the group on October 5th and offered to have a telephone conference during the meeting. Accordingly, nutrition education group members including Audrey Adler, Jennifer Anderson, Debra Palmer Keenan, Barbara Lohse and Nancy K. Lewis had a consultation session via telephone with Susan Welsh about ADA meeting and NC-1167 proposal renewal issues. Nancy K. Lewis updated and made suggestion to submit an American Dietetic Association (ADA) meeting conference grant proposal for September 16-19, 2006 meeting. Appropriateness and suitability of NC-1167 group to take part in the American Dietetic Association's Annual meeting were discussed. Feasibility of USDA conference grant proposal submission, possible industry support dollars, and identification of NC-1167 member who would take the leadership role in USDA conference grant proposal submission was also discussed. Kevin Fritsche asked to review about the timeline for year 2007-2012 proposal submission by NC-1167 to USDA. The group decided to have the draft proposal finished by December 2006. Members agreed to start writing the proposal during the 2006 Experimental Biology meeting and 2006 ADA meeting, and through out the year 2006. Review of our current proposal for any accomplishments to date and new directions were proposed. Jay Whelan and Ken Allen agreed to take a leadership role in writing NC-1167 renewal proposal for year 2007-2012. Based on suggestions from all members, Kevin Fritsche suggested that NC-1167 proposal renewal should be based on systematic review to determine whether the group adequately addressed the forms and the amount issues from the current cycle. Doreen Woodward suggested considering having the renewal proposal actually be interwoven among all projects rather than having parallel goals. Jay Whelan suggested reviewing how much collaborative effort has been made or will be made through out the current period and in the future proposal. Doreen Woodward (administrative advisor, Michigan State) updated the group about potential changes in CREES funding due to budget cuts and the possibility of future paradigm shifts. Thus, she suggested critical review of impacts and outcomes from our group in preparation for continuing support for our program. Doreen Woodward initiated an intense group discussion regarding how to measure impacts and how best to communicate impacts and outcomes using logic models. Doreen Woodward updated the group about the cancellation of the Annual Agricultural Experimental Station Director's meeting that was scheduled to be held at San Antonio (It was cancelled due to the hurricane). Doreen Woodward encouraged NC-1167 group members to communicate and update Agricultural Experimental Station Directors about group impacts and justifications of Multistate Research Programs. Mary A. Harris gave a 30 min research report on the CSU IFAFS DHA Gestational Duration study- The Intervention Phase. Jennifer Anderson gave a 30 min research report on the CSU IFAFS DHA Gestational Duration Study - The Nutrition Education Phase. Lively input, questions and suggestions were exchanged by all participants of the meeting. Breakout sessions were held at the Nutrition Department of CSU for Group I (Audrey Adler, Jennifer Anderson, Debra Palmer Keenan, Barbara Lohse, and Nancy K. Lewis) about the outreach and education goals. Group II (Allen, Baybutt, Broughton, Chapkin, Claycombe, Harris, Fritsche, Gallaher, and Whelan) met at the Hilton conference room to attend presentation of "Human equivalent dose modeling for omega-3 fatty acids" by Jay Whelan; followed by a discussion of animal (& cell culture) systems used to explore "form & amount" questions. Kevin Fritsche, Ken Allen and Jay Whelan indicated that n-3 PUFA - induced changes were still present in the studies in which US17 diets (with human equivalent levels of background PUFAs) were used. All members participated during the follow up group discussion regarding needs to establish standardization of n-3 PUFA DRIs. Business items: The following issues were discussed: formation of a writing committee for the renewal for next cycle, continuation of group focus (form and amount), EB meeting updates, deadline for ADA meeting conference. All members agreed to meet one day early at EB 2006 meetings to set up the framework for our NC-1167 proposal renewal (for year 2007-2012). Jennifer Anderson and Jay Whelan were asked to serve as speakers at the up coming 2006 ADA meeting in Hawaii. Jennifer Anderson and Jay Whelan also agreed to write ADA conference proposal to be submitted to ADA (arranged with consultation with ADA meeting organizer by Nancy K. Lewis). Kevin Fritsche and Kate Claycombe agreed to write NC-1167 final report for year 2002-2007 period. Kevin Fritsche suggested to update the NC-1167 litserve and to add Daniel Gallaher. Debra Palmer Keenan presented station report on behalf of the Nutrition education and outreach group (Audrey Adler, Jennifer Anderson, Debra Palmer Keenan, Barbara Lohse, and Nancy K. Lewis). Results from the web-based survey of registered dietitians revealed limited knowledge of forms and amounts of n-3 PUFA. Preparation of manuscript with emphasis on registered dietitians' strong interest in learning more to provide accurate information to consumers and other clients was indicated. Following this report, the group as a whole discussed the need to submit for a USDA/NRI conference grant by the December 13th deadline for year 2006. The first day's meeting was adjourned at 5:35 p.m. Jennifer Anderson and Jay Whelan met at 8:00 a.m. on Friday October 7th to finish drafting the ADA conference proposal. The second day meeting was called to order at 8:30 a.m. by the chair. Station Reports: The following member states gave research reports: North Carolina (2), Michigan, Texas, Missouri, Colorado, Tennessee, Wyoming, Nebraska, and Kansas. These reports elicited many questions and some lively discussion. More complete reports are attached to these minutes. Progress/Goals: Collaborative efforts of the basic science groups and nutrition education component were discussed. In the basic science part, use of US17 diet by multiple stations generated significant biological effects even with human equivalent doses of background n-3 PUFAs. Additional data will answer questions regarding the most important forms and amounts of dietary n-3 fatty acids for health maintenance and disease prevention which will be strengthened by documenting the effect in a number of different disease model or systems. Jay Whelan shared his review findings about how dietary n-3 PUFA pools change in vivo according to different dietary forms and amount of n-3 PUFA intake. Preparation of the manuscript from nutrition education component was indicated. Using identified progress (up to date) and future goals Debra Palmer Keenan initiated drafting ideas for USDA NRI proposal. Barbara Lohse agreed to serve as a secretary for year 2005-2006 and then chair for year 2006-2007. The meeting was adjourned at noon.

During 2005, work was begun on interpreting and documenting the results of the 190 surveys completed with Registered Dietitians (RDs) concerning their knowledge of omega-3 fatty acids, how they use this information with clients, and if and how they would like to receive additional education in the future on this topic. Additional analysis of results has been completed to further inform the strategies for the next stage of this project. Multistate partners from Nebraska, New Jersey, Pennsylvania State University, Rutgers, and Colorado State University are continuing data analysis of web-based and telephone surveys of dietitians in the four states. We have also begun initial steps in development of an omega-3 fatty acid educational website directed at nutrition practitioners. The coming year we plan to conduct qualitative assessments to identify dietitians responses to draft educational materials and proposed website layout. Basic Science components of this project: The Tennessee Station completed the developed and testing of a mathematical model for allometric scaling of polyunsaturated fatty acids (PUFA) between rodent models and humans. Their results provide a framework to improve the predictability of pre-clinical screening using experimental models as a surrogate for potential human responses. This would minimize false positives in animal experiments and unnecessary and expensive clinical trials that are, in part, based on these animal experiments. The Tennessee and Missouri Stations collaborated in an application of this novel allometric scaling model. Using rodent diets designed and formulated at Tennessee, the Missouri station conducted a feeding trial to examine the impact on host infectious disease resistance of supplementing EPA only to a basal rodent diet that mimicked current U.S. PUFA intake. The data suggest that providing mice a diet with extra EPA at levels equivalent to 1.4 en% will significantly diminish resistance against the food-borne bacterial pathogen, Listeria monocytogenes. The others Stations focused on investigating the biological activity of SDA (18:4n-3), a novel omega-3 fatty acid that we propose may serve as a land-based alternative source for the long-chain n-3 fatty acids found in marine and fish oil sources, EPA (20:5n-3) and DHA (22:6n-3). The Michigan Station reported that preadipocytes secrete significantly higher levels of LPS-induced IL-6 compared to differentiated adipocytes in murine cell lines and in primary cells that were isolated from mouse adipose tissues. Also, we showed that both EPA and SDA decrease LPS-induced IL-6 mRNA expression and IL-6 secretion significantly in adipose MCS cells and in 3T3-L1 preadipocytes. The Colorado station reported that SDA significantly increased only EPA in reprodutive tissue, the conversion of SDA to DHA appears to be limited by the requisite delta-6 desaturase that is proposed to be a part of the delta-4 desaturation step. Only at the highest level (2 en%) did SDA reduce uterine PGE2 production or fetal membrane PGF2alpha production in relation to LnA, and SDA does not affect uterine PGF2alpha or fetal membrane PGE2 production differently than does LnA. Given findings that lower levels of SDA were not any more effective in increasing tissue percent EPA or diminishing PG production than were higher levels of LnA, and since LnA and SDA did not differ in their effects on MMP-2 and MMP-9, further research is necessary to justify the use and doses of SDA supplements that are necessary to suppress PG production, prevent preterm PROM, and extend gestational length. The Wyoming Station conducted the first study examining the effect of dietary SDA and its effect on ovulation. When present in the diet of rats at only 0.6 wt% there was an enhancement in ova release. The North Carolina Station investigated the modulatory effects of unoxidized and oxidized PUFA on the production of several inflammatory responsive, pro-atherogenic mediators and their upstream activated transcription factors in LPS-stimulated macrophages. U937 cells were differentiated into macrophages by stimulation with PMA, incubated with 50 µM unoxidized and 15-LO oxidized arachidonic acid (AA), eicosapentaenoic acid (EPA), and docosahexaenoic acid (DHA) for 24h, and then stimulated with LPS (1¼g/ml) for 48-72h. The unoxidized PUFA produced no significant differences in LPS-induced IL-1² and IL-6 concentrations but decreased significantly the amount of mCRP compared to control, with unoxidized EPA and DHA being significantly more potent agents than unoxidized AA. All three oxidized PUFA significantly decreased the amount of LPS-induced IL-1², IL-6, and mCRP, with oxidized EPA being more potent than oxidized DHA and AA. Also, the LPS-induced activation of NF-ºB, C/EBP-², and STAT-3 was changed by to a greater extent by the oxidized PUFA than the unoxidized PUFA. The Texas Station investigated dietary perturbation of caveolae lipid microdomains and their subsequent altering of protein functionality. Caveolae isolated from mice fed n-6 (control) or n-3 PUFA enriched diets were characteristically enriched in cholesterol and sphingomyelin. n-3 PUFA reduced cholesterol and caveolin-1 content in the caveolae fraction by 46% and 53%, respectively. Concomitantly, localization of caveolae resident signaling proteins, H-Ras and eNOS, was decreased by 45% and 52%, respectively, and H-Ras activation was reduced. In contrast, localization of non caveolae proteins, K-Ras and clathrin was unchanged. These findings suggest that caveolae can be targeted in vivo by lipid modifying dietary agents and highlight the potential for caveolae to be exploited for therapeutic applications.

Al-Numair, Khalid and Lewis, Nancy M. 2005. Omega-3 fatty acid consumption and food sources differ among elderly men living in coastal and internal regions of Saudi Arabia. Pakistan Journal of Nutrition 4(2):106-111. J. Series No. 14308. Ritter-Gooder, P. and Lewis, Nancy M. 2005. Validity and reliability of a quantitative food frequency questionnaire measuring N-3 fatty acid intakes in cardiac patients in the rural Midwest. Journal of the American Dietetic Association 105:A16. Ritter-Gooder, P. Validity and reliability of a quantitative food frequency questionnaire measuring omega-3 fatty acid intakes in cardiac patients in the Midwest. M.S. Thesis, University of Nebraska. 2005. Troxell, H., Anderson, J., Auld, G., Marx, N., Harris, M.A., Reece M.S. and Allen, K.G.D. Omega-3 for Baby and Me: Material Development for a WIC intervention to Increase DHA Intake During Pregnancy. Maternal and Child Health Journal 9: 189-197, 2005. Lohse B, Shafer KJ, Theory-driven n-3 polyunsaturated fatty acid education delivered by written correspondence and problem-based approaches. Nutrition Research. (In Press). Jones, L. and Whelan, J., Human Equivalent Dose Modeling for Omega-3 Fatty Acid Supplementation in C57BL/6J Mice. 2005. FASEB J. 590.3A. Irons, R and Fritsche K.L. (2005) Omega-3 polyunsaturated fatty acids impair in vivo interferon-gamma responsiveness via diminishing receptor signaling. J Infect Dis. 191(3): 481-6. Irons, R., Pinge-Filho, P., Fritsche, K. (2005) Dietary (n-3) polyunsaturated fatty acids do not affect the in vivo development and function of Listeria-specific CD4+ and CD8+ effector and memory/effector T cells in mice. J. Nutr. 135: 1151-1156. Fritsche, K., Irons, R., Pompos, L., Janes, J., Zheng Z. and Brown, C. (2005) Omega-3 polyunsaturated fatty acid impairment of early host resistance against Listeria monocytogenes infection is independent of neutrophil infiltration and function. Cell Immunol (in press) Perez, M.A., Hansen, R.A., Harris, M.A. and Allen, K.G.D. Dietary docosahexaenoic acid alters pregnant rat reproductive tissue prostaglandin and matrix metalloproteinase production. J. Nutr. Biochem. In Press 2005. Rose, Janelle R. Dietary stearidonic acid and pregnant rat uterine and fetal membrane mediators of premature delivery. Masters degree Thesis, Colorado State University. Fall Semester 2004. Perez, Mark A. Dietary docosahexaenoic acid alters pregnant rat reproductive tissue prostaglandin and matrix metalloproteinase production. Masters degree Thesis, Colorado State University. Fall Semester 2005. Krugman, J, Rowe T, Tonso E, Tonso T, Driscoll M, Clement B, Anderson L, Hirschi K, Rule DC, Culver B, and Broughton KS. 2005 Effect of stearidonic acid on eicosanoid metabolism and ovulation. FASEB J. 19:A1007 (590.2) Harkins JM, Moustaid-Moussa N, Chung YJ, Penner KM, Pestka JJ, North CM, Claycombe KJ. Expression of interleukin-6 is greater in preadipocytes than in adipocytes of 3T3-L1 cells and C57BL/6J and ob/ob mice. J Nutr. 134(10):2673-2677, 2004 Ciubotaru, I., Potempa, L.A., and Wander, R.C. 2005. Production of modified C-reactive protein in U937-derived macrophages. Accepted Society Experimental Biology and Medicine. Lee, Y.S. and Wander, R.C. 2005. Reduced effect on apoptosis of 4-hydroxyhexenal and oxidized LDL enriched with n-3 fatty acids from postmenopausal women. J Nutr. Biochem. Apr 16(4): 213-221. Hall, J.A., Henry, L.R., Jha, S., Skinner, M.M., Jewell, D.E., and Wander, R.C. 2005. Dietary (n-3) fatty acids alter plasma fatty acids and leukotriene B synthesis by stimulated neutrophils from healthy geriatric Beagles. Prostaglandins, Leukotrienes, Essential Fatty Acids. Nov 73(5):335-41. Irina Ciubotaru, Ph.D. 2005. The Anti-Inflammatory Effects of Eicosapentaenoic and Docosahexaenoic Acids found in Fish in vivo and in vitro. Thesis. D.W.L. Ma, J. Seo, L.A. Davidson, E.S. Callaway, Y.Y. Fan, J.R. Lupton and R.S. Chapkin. n-3 PUFA alter caveolae lipid composition and resident protein localization in mouse colon. Faseb Journal, April 2004, 10.1096/fj.03-0604fje, 18:1040-1042, 2004. D.W.L. Ma, J. Seo, K.C. Switzer, Y.Y. Fan, D.N. McMurray, J.R. Lupton and R.S. Chapkin. n-3 PUFA and membrane microdomains: a new frontier in bioactive lipid research. J Nutr Biochem 15: 700-706, 2004.