Project/Activity Title: N-3 Polyunsaturated fatty acids in human health and disease.
Period Covered: November 1, 2003 – November 1 -2004.
Date of this report: November 10, 2004.
Annual Meeting Dates:
November 4-5, 2004 -
October 6-7, 2005 –
September ??? 2006 –
conjunction with annual
Participants: Kenneth G.D. Allen (Colorado State University), Jennifer Anderson (Colorado State University), Jay Whelan (University of Tennessee), Nancy K. Lewis (University of Nebraska), Audrey Adler (Rutgers University), Debra Palmer Keenan (Rutgers University), Kevin Fritsche (University of Missouri), Barbara Lohse (Kansas State University), Richard Baybutt (Kansas State University), Kate Claycombe (Michigan State University), Robert Chapkin (Texas A & M), K. Shane Broughton (University of Wyoming), Doreen Woodward (Administrative Adviser, Michigan State University), Susan Welch (CSREES Representative, Washington, DC).
Members Absent: Rosemary Wander (
meeting was called to order by the chair, K.
Shane Broughton, at 1:15 pm, November 4, 2004 at the
Kevin Fritsche served as secretary for this meeting.
Introductions & Opening Remarks:
The meeting opened with a welcome from John Baker, Acting Director of Michigan Ag. Expt. Station. He gave a brief overview of the areas of focus for the MAES: Food & Health, Environmental Stewardship, Food, Family & Fiber, Microbial Food Safety, Food Security.
We were introduced to our new USDA rep., Susan Welch. She told about her background as Natl. Program Leader for Nutrition Education Leadership. She informed us that the USDA has two open positions and the staff has been working hard over the past few months with finalizing the Dietary Guidelines Advising Report. Also, she told us that the NRI competitive grants program may get a $20M boost from Congress next fiscal year. How much of that will go into either of the nutrition-related programs (i.e., 31.0 and 31.5) is unclear. This is the first year of the new policy at the NRI regarding funding larger grants (~$500,000) and for longer (4 yrrs). As a result, in the 31.0 program only 10 of 104 applications were funded with a budget that has remained at ~$4M for many years. The new program (31.5) area on nutrition and obesity (31.5) the budget is ~$8.5M. These projects are supposed to integrate basic science with education, outreach, and outcomes. Ten out of 89 applications were funded in this program area. Susan encouraged us to consider applying for a conference grant to support the activities of this Multi-state project.
Doreen Woodward (Ad. Advisor, MSU) introduced herself and shared several helpful handouts regarding the NIMS reporting system and how to fill out the SAES-422 Annual Reports. She also reminded us that our mid-project report would be reviewed this Spring.
A research seminar entitled, “Hepatic metabolism and its impact on fatty acid-regulated transcription factors” was presented by Donald Jump (Prof. Physiol., Biochem. & Mol. Biol., MSU). The presentation was followed by many questions and a lively discussion.
1. The FDA allowed Health Claims for food rich in EPA and DHA were reviewed and discussed.
2. IFT Symposium: Members were asked if there was interest in organizing another symposium on n-3 PUFA nutrition for the annual IFT meeting. Several members had previously (July 2003) participated in such an activity at the invitation of Bruce Watkins, an IFT and NC1167 member. No one expressed an interest in organizing another IFT symposium, in part, because no one present was an IFT member.
3. EB’05 Symposium: “N-3 PUFA, Transitioning Research to Education” will be on April 5,
4. Society for Nutrition Education (SNE): Members of the NC1167 committee will be
organizing a symposium at SNE’s upcoming meeting in
(Note: Dr. Lohse, assigned to NC1167 from the Kansas Agricultural
Experiment Station, moved to
October 1, 2003 to September 30, 2004, the Nutrition Education unit of NC 1167
developed a survey for registered dietitians to assess n-3 fat knowledge,
practices regarding n-3 fat advice with patients, and n-3 fat continuing
education venue preferences. Surveys
were administered using two procedures:
self-report web-based and telephone interview with interviewers
recording responses on a web survey modified from the self-report version.
Following this report the group as a whole discussed the strength of the evidence to support benefits for a long list of health conditions. We attempted to categorize each health condition * EPA&DHA linkage as either: strong (A), modest (B), and weak or inconclusive (C), none (D). The subcommittee agreed to email this listing for further consideration by the entire committee.
The first day meeting was adjourned at 5:35 p.m.
The second day meeting was called to order at 8:40 by the chair.
1. Chair for next year will be Kevin Fritsche; secretary will be Kate Claycombe.
(Kate understands that she will be the incoming chair for ’06 as per our newly agreed upon line of succession.)
2. Next year’s meeting will held in Fort Collins, CO and be hosted by Ken Allen and Jennifer Anderson, reps. from Colorado State University.
3. NC1167 Listserv: Debra Palmer Keenan wanted to know if she should maintain the listserv for this group. She did so last year, yet no one seemed to use it. We all agreed that we should keep it and USE IT. All committee members are to send her an email from our email address that we want on the listserv.
The following member states gave “brief” research
Lunch & Guest Speaker:
We worked through lunch and had an informative discussion with Gale Strasburg, acting chair of the Food Science & Human Nutr. Dept. at MSU, regarding the recent reorganization of Colleges at MSU and the ongoing search for a new chair.
Progress/Goals: Collaborative efforts of the basic science groups and nutrition education component were discussed. The collaboration has strengthened as the project progresses; the group is developing into a cohesive core of n3 fatty acid studies. Collaborative efforts are in experimental design, n-3 form and amount, investigations being done under a number of different experimental conditions and the education of nutrition professionals. The result will be a general answer to a fundamental question regarding the most important forms and amounts of dietary n3 fatty acids for health maintenance and disease prevention which is strengthened by documenting the effect in a number of different systems. In addition, the nutrition education component will inform nutrition professionals on these results for education of RDs and their clients. Opportunities for obtaining NRI funding by various subgroups within this group were discussed. Kate Claycombe received a round of applause and the gratitude of her colleagues for an outstanding job of hosting the meeting.
The meeting was adjourned at 1:45 pm.
Reporting Period: October 1st 2003 – September 30th 2004
Personnel: Kenneth G.D. Allen (PI) Mary A. Harris (CoI)
Paul Kim (GRA), Mark A. Perez (GRA), Rodney Hansen (GRA)
Report: Pregnant rats were fed human equivalent amounts (0.7 energy %) and enhanced amounts (2.0 energy %) of the n-3 fatty acids linolenic (LnA) and docosahexaenoic (DHA) at constant n-6 linoleic acid (2 by 2 factorial design) from conception through day 20 of gestation. At day 20 of gestation DHA significantly decreased both placenta and uterus PGE2 production, uterus PGF2α production, both placenta and uterus collagenase activity (matrix metalloproteinase 1, 8 and 13 activity), and placenta active, but not pro-, matrix metalloproteinase (MMP) 2 and 9 expression. When analyzed by factorial ANOVA the main effects were attributable to fatty acid and not to dose. We conclude that the form of dietary n-3 fatty acid – DHA as opposed to LnA – is the most important in suppressing these indicators of shortened gestation and risk of premature delivery. Form is more important than amount and enhanced LnA (2 energy %) is not effective.
Using an immortalized late pregnancy human myometrial cell line (MHM1-41) in culture we have examined Ca2+ entry in response to oxytocin. Cells were cultured in DMEM 10% fetal bovine serum (FBS) and grown to ~ 70% confluence. Myometrial cells were provided with either DHA or oleic acid (OA) as 2:1 fatty acid: BSA by inclusion in culture media. DHA and OA were provided at 100, 30 and 10 mM. Following fatty acid provision for 3 days cells were loaded with 5μM Fura-2 AM Ca fluorescence dye and washed cells incubated with 1.0 mM Ca2+ in fluorescence buffer (FBS and fatty acid free). Changes is intracellular calcium – [Ca2+]i - were monitored by 340 nm and 380 nm excitation and 510 nm emission in response to 5-25 nM oxytocin using a calcium fluorescence microscope system. Identical parallel plates were used for cell membrane preparations. DHA provided at 100mM and 30mM significantly depressed
[Ca2+]i by 80% and 55% respectively relative to equimolar OA controls. For the 10mM experiments DHA suppressed [Ca2+]i by 30% which approached significance (p<0.08). Fatty acid analysis of membranes (> 95% phospholipid) showed enrichment of membrane DHA from 2.5% (BSA control) to ~ 5% (30 mM DHA) and ~ 10% (100 mM DHA). These data suggest that DHA incorporation into myometrial cell membranes alters oxytocin mediated Ca2+ entry. Possible mechanisms include membrane structure, phospholipase C activity, inositoltriphosphate production or alterations in G-coupled systems.
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,
Allen, K.G.D., Bristow, S.J. and Yu, L. Hypolipidemic effects of modified psyllium preparations. J. Agric. Food Chem. 52: 4998-5003, 2004.
Troxell H, Anderson J, Auld G, Marx, N, Harris M, Reece M, Allen K. Omega-3 For Baby and Me: Material Development For a WIC intervention to Increase DHA Intake During Pregnancy. Maternal & Child Health Journal (in press)
Abstracts and presentations:
Harkins, J.M., Hazan, A.M., Allen, K., Penner,
K.M., Pestka, J.J. and Claycombe, K.J. Modulation of
IL-6 expression and secretion in adipose tissue in virto
and in vivo by n-3 fatty acids. Presented at Experimental Biology 2004 Meeting,
Rose, Janelle R. Dietary stearidonic acid
and pregnant rat uterine and fetal membrane mediators of premature delivery.
Masters degree Thesis,
Personnel: Kevin L. Fritsche (PI)
Report: We investigated the impact of diet n-3 PUFA from fish oil on IFNg receptor expression and function. We hypothesized that reduced receptor expression would be both necessary and sufficient for n-3 PUFA-mediated hypo-responsiveness to IFNg to occur. Such an effect would then lead to greater host susceptibility to L. monocytogenes infection. Using IFNg null mice (IFNg-/-) on a BALB/c background, we showed that high intake of n-3 PUFA could create an in vitro and in vivo state of IFNg hypo-responsiveness. Surprisingly, this effect occurred without a reduction in surface receptor expression. These findings suggested that n-3 PUFA impairment of the IFNg response might be mediated through changes within the cell (i.e., a receptor signaling pathway). We have recently demonstrated that IFNg-mediated phosphorylation of Stat1, an early and critical factor in the IFNg signaling pathway, was cut in half in resident peritoneal macrophages isolated from mice fed a diet rich in n-3 PUFAs. These data suggest that dietary n-3 PUFA impair in vivo interferon-gamma responsiveness via diminishing receptor signaling. Additional studies are planned to elucidate the underlying molecular mechanism(s) responsible for this aberrant signaling and to explore the possibility that other cytokine receptors may be similarly affected.
Zhang, M. and Fritsche K.L. (2004) Fatty acid-mediated inhibition of interleukin-12 production by murine macrophages is independent of peroxisome proliferator-activated receptor-g. Brit. J. Nutr. 91: 733-739.
Anderson M and Fritsche KL (2004) Dietary polyunsaturated fatty acids modulate in vivo, antigen-driven CD4+ T-cell proliferation in mice. J. Nutr. 134: 1978-1983.
Irons R, Pinge-Filho P, and Fritsche KL. (2004) Omega-3 polyunsaturated fatty acids impair in vivo interferon-gamma responsiveness via diminishing receptor signaling. J Infect Dis. (in press).
Irons, R. (2004) Fish oil severely impairs immunity to Listeria monocytogenes
without affecting the adaptive immune response. Ph.D. Dissertation,
Personnel: Kate Claycombe (PI)
Report: An increasing number of studies have shown that inflammation plays a major role in the development of cardiovascular diseases. IL-6, a proinflammatory cytokine, is secreted from variety of cell types such as activated leukocytes, endothelial cells, adipocytes and preadipocytes. We recently have shown 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. Using these data as bases, we tested whether n-3 fatty acids SDA (18:4, n-3) have anti-inflammatory effects on lipopolysaccharide (LPS)-induced IL-6 secretion in 3T3-L1 preadipocytes. Our data showed that SDA treatment of 3T3-L1 cells for 48 hrs significantly increase EPA levels (20:5, n-3) in the phospholipid fraction. We further showed that both EPA and SDA decrease LPS-induced IL-6 mRNA expression and IL-6 secretion significantly in 3T3-L1 preadipocytes. Mechanisms underlying the inhibitory effect of SDA and EPA on LPS-induced IL-6 mRNA and secretion are currently being investigated.
Harkins, JM, Moustaid-Moussa, N, Penner, KM, Pestka, JJ, Chung, YJ, North, CM, and 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. 134:2673-2677, 2004.
Harkins, JM, Whelan J, Jones L, Ken Allen, and Claycombe, KJ. Inhibitory effect of SDA and EPA on 3T3-L1 preadipocyte secretion of interleukin-6 (IL-6). In preparation, 2004.
Personnel: Nancy M. Lewis (PI)
Report: This research addresses Objective 3 of the current NC-1167 project. Dietary intakes of omega-3 fatty acids are influenced by geographic region and intakes of those living in the Midwestern United States typically do not meet current recommendations. The purpose of our research is to identify factors influencing omega-3 fatty acid intakes and to design effective nutrition education interventions to increase intakes. This year we established the reliability of a previously developed omega-3 fatty acid food frequency (FFQ) and we used the FFQ to evaluate the effectiveness of a nutrition education intervention in increasing omega-3 fatty acid intakes in heart patients. Thirty-six adults were divided into two groups. Both groups received a 2-hour workshop to increase cognitive and behavioral strategies for increasing consumption of omega-3 fatty acids. Group 1 received a follow-up one-on-one discussion with a registered dietitian to identify and discuss personal pros and cons of changing eating behaviors. Group 2 received a follow-up telephone call. Both Groups doubled their n-3 intakes from baseline to 1 month post. Group 1 maintained >80% of the increase at 2 months post while group 2 maintained only 35 % of the increase at 2 months post. We conclude that inclusion of decisional balance in this nutrition education intervention increased long term maintenance of omega-3 fatty acid intakes.
Multistate partners from Kansas State University (now Pennsylvania State University), Rutgers University and Colorado State University conducted both a telephone and a web-based survey of practicing dietitians to assess knowledge of n-3 fatty acids and continuing education needs in this area. Data from this survey will be used in the next step of this project which is to design a nutrition education intervention with dietitians.
IMPACT: Identification of components of n-3 fatty acid nutrition education interventions that increase effectiveness of the interventions will increase the credibility of nutrition educators and will result in more significant health outcomes for consumers.
Sindelar, Carrie A., Lewis, Nancy M., Scheerger, Sarah B., Plugge, Sheri L., Eskridge, Kent M., and Wander, Rosemary C. 2004. Serum lipids of physically active adults consuming omega-3 fatty acid-enriched eggs or standard eggs. Nutrition Research 24:731-739.
Heidal, Kimberly, Lewis, Nancy M. 2004. Omega-3 fatty acid nutrition education resources. Journal of Nutrition Education and Behavior 36 (4):209-210.
Al-Numair, Khalid and Lewis, Nancy M. 2004. Omega-3 fatty acid intake and
incidence of nonfatal myocardial infarction differ between coastal and internal
Al-Numair, Khalid .S. 2004. Comparison
of the intake of omega-3 fatty acids and its relation to the incidence of non
fatal myocardial infarction in two samples from different geographical
locations in Saudi Arabia. Ph.D. Dissertation,
Personnel: Audrey Adler (co-PI), Debra Palmer Keenan (co-PI)
year 2004, the protocol and two survey questionnaires developed in fiscal year
2003 were implemented to query 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.
The research protocol was approved by the Rutgers Institutional Review
Board, as well as those in
A web-based version of the survey questionnaire was
administered by researchers at
All data, from both the telephone and web-based surveys,
were forwarded to
Analyses will continue in fiscal year 2005 to further inform the intervention strategies and materials that will be developed beginning this year.
Personnel: Rosemary C. Wander (PI)
Report: The role of n-3 PUFA in Cardiovascular
Disease: Cardiovascular disease (CVD) represents the leading cause of mortality
In a recent human study conducted by the NC Station, thirty healthy women taking hormone replacement therapy were randomly divided into three groups and supplemented for five weeks with 14g/d safflower oil (SO), 7g/d of both safflower oil and fish oil (LFO), or 14g/d fish oil (HFO). The concentration of serum C-reactive protein (CRP) and interleukin 6 (IL-6) in plasma and in cell culture supernatant collected from 24-h lipopolysaccharide (LPS)-stimulated whole blood was measured. In addition, a panel of lipid markers was assessed. Fish oil supplementation significantly decreased CRP and IL-6 compared to SO, with a greater effect in the LFO than the HFO groups. Plasma triacylglycerol (TG) and the TG/HDL-C ratio were significantly lower in the HFO compared to the SO group (Ciubotaru et al, 2003).
Low density lipoprotein (LDL) from each group was also obtained after 5-wk supplementation and oxidized with CuSO4. The concentration of cholesteryl ester hydroperoxide (CEOOH) and the formation of conjugated dienes in the oxidized LDL (oxLDL) were measured. Additionally, apoptotic events in response to oxidized LDL derived from the three supplementation groups were measured in U937 monocytes. There was no significant difference among the treatment groups with respect to CEOOH concentrations and conjugated diene formation in oxLDL. However, the percent of apoptotic U937 monocytes was approximately 40 % lower upon incubation with oxLDL obtained from the HFO-supplemented group compared to the SO-supplemented group (Lee and Wander, in press). Previously, we have also shown similar protective effects on apoptosis of U937 cells which were incubated with oxidized EPA/DHA enriched LDL compared to non-EPA/DHA enriched LDL (Wu et al, 2002). These results suggest that dietary fish oil may decrease the risk for cardiovascular disease through the modulation of plasma lipids, inflammatory markers, and apoptosis. Therefore, the objectives of our NC1167 research project are to take these observation further, and using in vitro models to elucidate the mechanism that drives the anti-atherogenic and anti-apoptotic properties of fish consumption.
Lee Y.S. and Wander R.C. Reduced effect on apoptosis of 4-hydroxyhexenal and oxidized LDL enriched with n-3 fatty acids from postmenopausal women. J Nutr Biochem. 2004. In Press.
Ciubotaru I, Lee YS, Wander. Dietary fish oil decrease C-reaction protein, interleukin-6, and triacylglycerol to HDL-cholesterol ratio in postmenopausal women on HRT. J Nutr Biochem. 2003 Sep;14(9):513-21.
Toobert DJ, Glasgow RE, Strycker LA, Barrera M Jr, Radcliffe JL, Wander RC, Bagdade JD. Biologic and quality-of-life outcomes from the Mediterranean Lifestyle Program: a randomized clinical trial. Diabetes Care. 2003 Aug;26(8):2288-93.
Hall JA, Tooley KA, Gradin JL, Jewell DE, Wander RC. Effects of dietary n-6 and n-3 fatty acids and vitamin E on the immune response of healthy geriatric dogs. Am J Vet Res. 2003 Jun;64(6):762-72.
Manns PJ, Williams DP, Snow CM, Wander RC. Physical activity, body fat, and serum C-reactive protein in postmenopausal women with and without hormone replacement. Am J Hum Biol. 2003 Jan-Feb;15(1):91-100.
Personnel: Jay Whelan (PI)
This project is addressing the following two objectives: (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; (2) use experimental diets in animal studies to examine
dietary levels of n-3 PUFA that are achievable in human diets, based on human
equivalent amounts (allometric scaling) in rodent
models. To date, we have established a
mathematical model predicting allometric scaling for
dietary polyunsaturated fatty acids between rodent models and humans and began
testing this mathematical model using C57BL/J6 mice. We compared allometric
scaling based on a number of conversion factors: body weight, body weight to
the 3/4 power (Max Kleiber’s conversion formula) and
caloric intake in the form of % energy.
Using direct comparisons of each formula, allometric
scaling based on caloric intake not only linearized
the relationship of nutrient intakes across species with different body sizes,
but the conversion was more closely aligned with the daily reference intakes
for humans for each of the nutrients identified in the mouse diet. Following adjustment and validation of the
mathematical model, a series of experiments were performed in C57BL/J6 mice
comparing dietary alpha-linolenic acid (
Whelan, J. and McEntee, M.F. 2004. Dietary N-6 Polyunsaturated Fatty Acids and Intestinal Tumorigenesis. J. Nutr. 134 (Dec issue), (In press).
J., McEntee, M.F. and Baek,
S.J. 2004. Dietary Polyunsaturated Fatty Acids, Eicosanoids, and Intestinal Tumorigenesis. In: Bioactive Lipids in Cancer. Carcinogenic and Anticarcinogenic Food
Components (Ed., Sikorski, Z. E.) CRC Press.
J. and McEntee, M.F. 2003. NSAIDs, Prostaglandins and APC-Driven Intestinal Tumorigenesis. In: Cyclooxygenase-2 Blockade in Cancer
Prevention and Therapy
(Ed., Harris, R.E.) Humana
Personnel: Robert S. Chapkin (PI)
Report: This project addresses the following two objectives: (1) to determine how n-3 PUFA uniquely modulate surface receptor protein function and T-cell responsiveness by altering membrane microdomain lipid composition; (2) to determine how n-3 PUFA alter intestinal membrane microdomain lipid composition and resident protein signaling. Our data indicate that (i) the anti-inflammatory properties of dietary n-3 PUFA are the result of a coordinated direct effect on T-cell proliferation and activation induced cell death (AICD); (ii) at least two co-receptor molecules (CD28 and CTLA-4) providing positive and negative signals, respectively, are involved in regulating this diet-induced effect; (iii) the propensity of n-3 PUFA to enhance the polarization of T-cells toward a Th1-AICD prone population suggest that dietary n-3 PUFA may act differently on distinct T-cell subsets; and (iv) the dietary n-3 PUFA modification of T-cell lipid raft microdomain structure, which is associated with alterations in the assembly of signal transduction complexes and the induction of AICD, must be sustained in culture using homologous mouse serum to observe the T-cell functional changes.
We have shown that the balance between colonic epithelial cell proliferation and apoptosis can be favorably modulated by dietary n-3 PUFA, conferring resistance to toxic carcinogenic agents. This is significant because the typical Western diet contains 10 to 20 times more n-6 than n-3 PUFA. In a major step toward developing a unifying mechanistic hypothesis addressing why n-3 PUFA suppress colon cancer compared to n-6 PUFA (the major dietary form of PUFA in the U.S. diet), we demonstrate for the first time that n-3 PUFA feeding is capable of markedly altering lipid-protein composition of colonic caveolae microdomains, thereby selectively modulating localization and function of caveolar proteins. In addition, our in vivo dietary intervention study carries significant biological relevance, compared with commonly used extreme pharmacological perturbations. Overall, our findings highlight a novel modality by which n-3 PUFA influence membrane microorganization, thereby modulating biological responses.
K.C. Switzer, D.N. McMurray, J.S. Morris and R.S. Chapkin. Dietary n-3 polyunsaturated fatty acids selectively promote activation-induced cell death in T-lymphocytes. Journal of Nutrition 133:496-503, 2003.
Y.Y. Fan, D.N. McMurray, L.H. Ly and R.S. Chapkin. Dietary n-3 polyunsaturated fatty acids remodel mouse T cell lipid rafts. Journal of Nutrition 133:1913-1920, 2003.
L.K. Bancroft, J.R.
M.H. Hong, R.S. Chapkin,
Y.Y. Fan, T.E. Spencer, N. Wang, M.P. Moyer and R.S. Chapkin. Chemopreventive n-3 fatty acids activate RXRa in colonocytes. Carcinogenesis 24:1-8, 2003.
T.V. Apanasovich, S. Sheather,
J.R. Lupton, N.
D.W.L. Ma, J. Seo,
L.M. Sanders, C.E. Henderson, M.Y. Hong, R. Barhoumi, R.C. Burghardt, R.J. Carroll, N.D. Turner, R.S. Chapkin and J.R. Lupton. Pro-oxidant environment of the colon compared to the small intestine may contribute to greater cancer susceptibility. Cancer Letters 208:155-161, 2004.
K.C. Switzer, Y-Y. Fan, N. Wang, D.M. McMurray and R.S. Chapkin. Dietary n-3 polyunsaturated fatty acids promote activation-induced cell death in the Th1-polarized murine CD4+ T-cells. Journal of Lipid Research, 45:1482-1492, 2004.
L.A. Davidson, D.V. Nyuyen, R.M. Hokanson, E.S. Callaway, R.B. Isett, N.D. Turner, E.R. Dougherty, J.R. Lupton, R.J. Carroll and R.S. Chapkin. Chemopreventive n-3 polyunsaturated fatty acids reprogram genetic signatures during colon cancer initiation and progression in the rat. Cancer Research 64:6797-6804, 2004.
Y.Y. Fan, L.H. Ly, R. Barhoumi, D.N. McMurray and R.S. Chapkin. Dietary docosahexaenoic acid suppresses T-cell protein kinase C-theta lipid raft recruitment and interleukin-2 production. Journal of Immunology (In press).
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. Journal of Nutritional Biochemistry (In press).
L.M. Sanders, C.E. Henderson, M.Y. Hong, R. Barhouim, R.C. Burghardt, N. Wang, C.M. Spinka, R.J. Carroll, N.D. Turner, R.S. Chapkin and J.R. Lupton. Enhancement of reactive oxygen species by dietary fish oil and attenuation of antioxidant defenses by dietary pectin coordinately heightens apoptosis in the rat. Journal of Nutrition (In press).
D.W.L. Ma, R.H. Finnell, L.A. Davidson, E.S. Callaway, O. Spiegelstein, J.A. Piedrahita, J.M. Salbaum, C. Kappen, B. Weeks, J. James, D. Bozinov, J.R. Lupton and R.S. Chapkin. Folate transport gene inactivation in mice increases sensitivity to colon carcinogenesis. Cancer Research (In press).
L.H. Ly, R. Smith, R.S. Chapkin and D.N. McMurray. Dietary n-3 polyunsaturated fatty acids suppress splenic CD4+ T-cell function in IL-10(-/-) mice. Clinical and Experimental Immunology (In press).
Personnel: K. Shane Broughton (PI)
Report: The objective of this work is to determine the effect n-3 PUFA consumption at achievable levels on compounds and expression of enzymes involved in ovulation: Previous studies in rats demonstrated that dietary intake of the n-3 PUFA stearidonic acid (SDA) at 0.3en% resulted in a 120% elevation in cyclooygenase expression in ovarian tissue. Current studies are in progress to compare the effect of three different dietary levels of SDA (0.15, 0.3, and 0.6 en%) and eicosapentaenoic acid (EPA)( 0.3 en%) on cyclooxygenase 1 (COX-1) expression, prostaglandin E2 and F2a biosynthesis, and ovulation. Following feeding from 15 days of gestation until 24 days of female rat pup age, Sprague Dawley rats are maintained on a diet that is isocaloric with a variation in the n-3 PUFA content. Following induction of ovulation with pregnant mares serum gonadotropin followed by human chorionic gonadotropin, female rats are euthanized and the eggs in the oviduct are assayed along with prostaglandin production and ovarian COX-1 expression.
None related to this research.
Abbreviated State (Station) Reports for NC-1167 Committee
Kenneth G.D. Allen (PI)
Mary A. Harris (CoI)
Department of Food Science and Human Nutrition
We investigated the importance of both the form and
the amount of dietary n-3 polyunsaturated fatty acids (PUFA) on indices of
gestational length and premature delivery in rats. We constructed rat diets to
provide relatively constant linoleic acid (LA) which
ranged from 5.4 to 6.3 energy (en) %, containing either 0.7 en% n-3 PUFA or 2.0
en % PUFA as linolenic acid, docosahexaenoic,
or SDA. At day 20 of gestation uterine prostaglandin (PG) E2 and F2α
synthesis rates were both significantly depressed approximately 50% by both 0.7
en% and 2.0 en% DHA, but not by 2.0 en% LnA.
Placental collagenase activity (matrix metalloproteinase (MMP) 1, 8 and 13 activities) was 50%
depressed by 0.7 en % and 2.0 en% DHA and by 2.0 en% LnA.
Placental active MMP-2 and MMP-9 expression were 40% suppressed by 0.7 en% and
2.0 en% DHA. Decreases in PG synthesis and MMP activity and expression are
associated with increased gestational length and reduced incidence of premature
delivery. Replacing LnA with DHA in diets, without
increasing n-3 PUFA intakes from current
K. Shane Broughton (PI)
Nutrition/Dept. of Family and Consumer Sci.
The influence of different forms of n-3 fatty acids, 2.2 energy% of the n-3 PUFA stearidonic acid (SDA) and eicosapentaenoic acid (EPA) and conjugated linoleic acid (CLA on ovulation and the expression of enzymes involved in ovulation. Sprague Dawley rats were maintained on a diet that was isocaloric with a variation in the n-3 PUFA source, or consumed a control diet or a diet enriched in conjugated linoleic acid (CLA). Following induction of ovulation with pregnant mares serum gonadotropin followed by human chorionic gonadotropin, the female rats were euthanized and the eggs in the oviduct were assayed along with prostaglandin production and ovarian cyclooxygenase 1 and 2 (COX-1 and COX-2) expression. While egg release did not vary at this level of dietary lipid modification, COX-1 expression was increased by 120% with SDA ingestion over control or EPA ingestion. Conversely, with CLA ingestion, COX-2 expression was reduced to 68% of that seen by the control and EPA animals. Prostaglandin release in correlation with COX expression is under analysis.
Kate J. Claycombe
Dept. Food Science and Human Nutrition
An increasing number of studies have shown that inflammation plays a major role in the development of cardiovascular diseases. IL-6, a proinflammatory cytokine, is secreted from variety of cell types such as activated leukocytes, endothelial cells, and adipocytes and is a potent inducer of hepatic C-reactive protein (CRP), one of the most sensitive markers of CVD risk. Recent studies also have demonstrated that adipose tissue releases large amounts of CRP activator, IL-6, in vivo. We tested whether murine 3T3-L1 preadipocytes secret significant levels of IL-6 when compared to differentiated 3T3-L1 adipocytes and whether anti-inflammatory n-3 fatty acids such as EPA (20:5, n-3) and SDA (18:4, n-3) inhibit lipopolysaccharide (LPS)-induced increases in IL-6 secretion in murine 3T3-L1 preadipocytes. Our data showed that preadipocytes secrete significantly higher levels of IL-6 than adipocytes, and treatment of preadipocytes with both EPA and SDA decreases LPS-induced IL-6 secretion significantly by approximately 70%. We conclude that one way by which n-3 fatty acids can reduce cardiovascular disease risk may be via down regulating IL-6 in the preadipocytes.
Kevin Fritsche (PI)
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 LCn-3 PUFA, e.g. fish oil, as illustrated by the reduced bacterial clearance we have observed in the liver and spleen 3 to 4 days post-infection. 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. 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 energy%). Current studies are investigating the level of LCn-3 PUFA and the impact of various n-3 PUFA forms (alpha-linolenic, stearidonic, EPA and DHA) on host infectious disease resistance.
Nancy M. Lewis (PI)
Department of Nutritional Sciences and Dietetics
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
During this past year, NC-1167 partners from Kansas, Colorado, New Jersey and Nebraska have worked as a team to develop a detailed questionnaire for assessing omega-3 knowledge and practices of dietitians which was 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.
Barbara Lohse PhD, RD, LD
Department of Human Nutrition
Kansas State University Extension Agents in the
College of Human Ecology have been teaching consumers about omega-3 fatty acids
using the Lesson, “Omega-3s: Fats you
Should and Can Eat,” written by Barbara Lohse (Knous). A second
omega-3 problem that encouraged menu planning was developed and the omega-3
Debra Palmer Keenan
Department of Nutritional Sciences
Fax: (732) 932-6522
. An open
ended, qualitative survey prototype was developed to collect preliminary
information on what Registered Dieticians (RDs) know
about omega-3 fatty acids, what they believe their clients know, what they
teach their clients, and what kind of educational opportunities would be most
useful to them to learn more. A convenience sample of ten RDs
Jay Whelan (PI)
Department of Nutrition
Studies are evaluating the effect of different n-3 fatty acids, both form (source) and amount, on tissue functions and correlating these effects with changes in putative biomarkers relevant to health promotion and disease prevention. 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. We are currently establishing a mathematical models predicting allometric scaling for dietary polyunsaturated fatty acids between rodent models and humans and testing this mathematical model using C57BL/J6 mice.
Following adjustment and validation of the mathematical
model, a series of experiments will be performed in the same murine model where we will be comparing dietary alpha-linolenic acid (
Jennifer E.L. Anderson, PhD, RD
Department of Food Science and Human Nutrition
Fax: (970) 491-7252
this past year the NC 1167 partners from