2006 Annual Report 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? Why does it matter? Cardiovascular disease is the leading cause of death and disability in Americans. The risk of developing cardiovascular disease increases dramatically with age. Major risk factors for heart disease and stroke include age, male gender, high blood pressure, diabetes, cigarette smoking and altered levels of lipoproteins (particles in the blood that contain cholesterol, proteins and other fats, such as triglyceride), increased blood levels of total cholesterol (>240 mg/dl), increased levels of low-density lipoprotein (LDL) cholesterol (>160mg/dl) and decreased levels of high-density lipoprotein (HDL) cholesterol (<40 mg/dl in men and <50 mg/dl in women). LDL - the “bad” cholesterol, deposits cholesterol into tissues in the body whereas HDL is important for cholesterol removal, a process known as reverse cholesterol transport. Being obese markedly increases the prevalence of diabetes, high blood pressure, increased LDL and decreased HDL cholesterol levels. Emerging risk factors for cardiovascular disease include increased levels of small, dense LDL, decreased levels of large HDL particles, elevated levels (>30 mg/dl) of lipoprotein (a), a genetically determined lipoprotein, blood markers of inflammation and increased white cell activity such as increased levels of C-reactive protein (CRP), lipoprotein associated phospholipase A2 (LpPLA2), fibrinogen and serum amyloid A protein. Prevention of heart disease and stroke by treating the risk factors is by far the most effective treatment strategy. Dietary modification remains the cornerstone of the prevention of cardiovascular disease. Although we know that diet can play an important role in maintaining cardiovascular health by lowering LDL cholesterol, our understanding of the absolute impact of individual dietary components in this process is far from complete. Although we know that diet can play an important role in maintaining cardiovascular health, our understanding of the absolute impact of individual dietary components in this process is far from complete. Similarly, our understanding of diet/gene interactions, especially as they contribute to variability in response to dietary modification, is in its infancy. The focus of this project, which encompasses the work of the Lipid Metabolism and Cardiovascular Nutrition laboratories is to investigate dietary strategies intended to maintain cardiovascular health, develop a screening tool for the early identification of individuals at high-risk of developing cardiovascular disease, and expand our understanding at a biochemical level of how diet alters measures of cardiovascular disease risk. Our research focuses on the types of dietary fats, carbohydrates, protein and fiber, as well as cholesterol, and their relationships to cardiovascular risk, obesity, dementia, plasma lipoproteins and their subspecies and inflammatory markers. By conducting this research we will provide the scientific community with solid research data that justify specific recommendations for dietary modification. These recommendations aim to favorable impact plasma lipoprotein and glucose levels, inflammatory markers, blood pressure, weight and overall cardiovascular disease risk. Research is also conducted to assess the efficacy of selectively bred and genetically modified vegetable oils, new sources of plant sterols, and soy protein on cardiovascular disease risk factors. Scientists are working to determine the impact of diet and genotype on cholesterol homeostasis and cardiovascular health in the Framingham Offspring Study; and to relate diet-induced changes in cardiovascular disease risk factors to functional changes in gene expression and atherosclerotic lesion formation in animal models and cell culture. The objectives are to test novel dietary interventions aimed at improving biological markers of cardiovascular health, defining diet/gene interactions that might mitigate nutrition interventions and determine more accurate approaches to identify high-risk individuals for early and intensive dietary intervention to maintain cardiovascular health with advancing age. The results of this work are relevant to the general United States (US) population and relate to the following program components within the National Program 107 - Human Nutrition: 3. Nutrition Monitoring; 5. Health Promoting Properties of Plant and Animal Foods; 6. Prevention of Obesity and Disease: Relationship between Diet, Genetics, and Lifestyle; and 7: Health Promoting Intervention Strategies for Targeted Populations. 2.List by year the currently approved milestones (indicators of research progress) The objectives of the Lipid Metabolism Laboratory are: 1. To determine the effect of altering dietary composition by restricting carbohydrates, fats, glycemic load, or total calories on body weight, and cardiovascular risk factors in overweight and obese subjects in the free-living state. 2. To determine the effect of altering fat and carbohydrate level as well as carbohydrate type or glycemic index and load on weight loss and cardiovascular risk factors in overweight or obese subjects, under controlled, ad libitum, and isoweight conditions. 3. To determine the effect of the standard 2005 Pyramid Diet versus a low glycemic load Pyramid Diet on weight loss and heart disease risk factors in overweight and obese subjects in the free-living state. 4. To develop and test an interactive web-based program to provide an optimal diet and exercise program for middle-aged and elderly subjects for weight and heart disease reduction. 5. To observe the interactions of nutritional factors, especially intake of calories, types of fat, types of carbohydrate, level of physical activity, and different genetic factors on lipoprotein subspecies, obesity, metabolic syndrome, inflammatory markers, and heart disease risk in overweight and obese subjects and subjects with premature cardiovascular disease as compared to age- and gender-matched control subjects within populations. 6. To determine the in vitro and in vivo effects of dietary fatty acids, cholesterol, carbohydrates, hormone levels, hormonal replacement, B vitamins, cholesterol biosynthesis inhibition and cholesteryl ester transfer protein inhibition on lipoprotein metabolism and gene expression, and inflammation in human liver cells (HepG2) and in human subjects under metabolic ward conditions using stable isotopes. The objectives of the Cardiovascular Research Laboratory are: 1. Examine the efficacy of selectively bred and genetically modified vegetable oils and new sources of plant sterols on cardiovascular disease risk factors in middle-aged and older hypercholesterolemic female and male subjects. 2. Determine the effect of soy protein on cardiovascular risk factors in middle-aged and older hypercholesterolemic female and male subjects. 3. Determine the impact of diet and genotype on cholesterol homeostasis and cardiovascular health in the Framingham Offspring Study – Cycle 6. 4. Relate diet-induced changes in plasma lipid and lipoprotein levels to functional changes in gene expression and atherosclerotic lesion formation in Golden-Syrian hamsters. Milestones: Lipid Metabolism Laboratory 2005 1. To determine the effects of markedly restricting either total fat or total carbohydrate, or increasing protein in the diet versus a more balanced approach of caloric restriction on weight loss and cardiovascular risk factors in obese subjects over a 1-year period. (Objective 1). 2006 1. To determine the effect of altering fat and carbohydrate level as well as carbohydrate type or glycemic index and load on weight loss and cardiovascular risk factors in overweight or obese subjects, under controlled, ad libitum, and isoweight conditions. (Objective 2). 2. To assess the interaction of nutritional factors, especially intake of calories, total fat, saturated fat, carbohydrate and sugar, level of physical activity and different genetic factors on obesity, insulin resistance, inflammatory markers, lipoprotein subspecies and heart disease risk in overweight and obese subjects with premature cardiovascular disease as compared to age- and gender-matched control subjects within populations (Objective 5). 2007 1. To determine the in vitro and in vivo effects of dietary fatty acids, cholesterol, and different carbohydrates, hormones, hormonal replacement, B vitamins, cholesterol biosynthesis inhibition and cholesteryl ester transfer protein inhibition on lipoprotein metabolism and gene expression, and inflammation in human liver cells (HepG2) and in human subjects under metabolic ward conditions using stable isotopes. (Objective 6) 2008 1. To determine the effects of the standard 2005 Pyramid Diet versus a low glycemic load Pyramid Diet on weight loss and heart disease risk factors in overweight and obese subjects. (Objective 3) 2009 1.To develop and test an interactive web-based program to provide an optimal diet and exercise program for middle-aged and elderly subjects for weight loss and heart disease risk reduction. (Objective 4) Milestones: Cardiovascular Nutrition Laboratory 2005 1. Determine the effect of genetically modified and selectively breed soybean oils on cardiovascular disease risk factors. Objective 1 2. Determine the effect of plant sterols with and without very long chain omega-3 fatty acids on cardiovascular disease risk factors. Objective 1 3. Determine the effect soybean derived foods on cardiovascular disease risk factors. Objective 2 4. Begin measures of surrogate markers of cholesterol homeostasis in case and matched control subjects. Objective 3 5. Begin to relate measures of cholesterol homeostasis to the incidence of cardiovascular disease and carotid stenosis. Objective 3 6. Determine the effect of dietary fatty acids on mRNA abundance of genes involved with the metabolism of low-density lipoprotein (LDL) cholesterol and high-density lipoprotein cholesterol in Golden Syrian Hamsters. Objective 4 2006 1. Complete studies on the effect of genetically modified and selectively breed soybean oils on cardiovascular disease risk factors. Objective 1 2. Complete studies on the effect of plant sterols with and without very long chain omega-3 fatty acids on cardiovascular disease risk factors. Objective 1 3. Complete studies on the effect soybean derived foods on cardiovascular disease risk factors. Objective 2 4. Continue work relating measures of cholesterol homeostasis to incidence of cardiovascular disease and carotid stenosis in case and matched control subjects. Objective 3 5. Begin to establish adult normal ranges for phytosterol, cholestanol and cholesterol precursor levels using samples collected during Cycle 6 of the Framingham Offspring Study. Objective 3 6. Continue determining the effect of dietary fatty acids on mRNA abundance of genes involved with the metabolism of low-density lipoprotein (LDL) cholesterol and high-density lipoprotein cholesterol in Golden Syrian Hamsters. Objective 4 7. Begin to determine the effect of dietary fatty acids on mRNA abundance of genes involved with the metabolism of low-density lipoprotein (LDL) cholesterol and high-density lipoprotein cholesterol in LDL knockout mice. Objective 4 2007 1. Complete work on relating measures of cholesterol homeostasis to incidence of cardiovascular disease and carotid stenosis in case and matched control subjects. Objective 3 2. Continue work to establish adult normal ranges for phytosterol, cholestanol and cholesterol precursor levels using samples collected during Cycle 6 of the Framingham Offspring Study. Objective 3 3. Begin work to define the relationship between phytosterol, cholestanol and cholesterol precursor levels and plasma lipid, lipoprotein and apolipoprotein levels. Objetive 3 4. Begin work to define the relationship between phytosterol, cholestanol and cholesterol precursor levels and selected dietary intake data (macronutrients, cholesterol, fatty acids, fiber and antioxidant supplements). Objective 3 5. Continue work determining the effect of dietary fatty acids on mRNA abundance of genes involved with the metabolism of low-density lipoprotein (LDL) cholesterol and high-density lipoprotein cholesterol in Golden Syrian Hamsters. Objective 4 6. Continue studies to determine the effect of dietary fatty acids on mRNA abundance of genes involved with the metabolism of low-density lipoprotein (LDL) cholesterol and high-density lipoprotein cholesterol in LDL knockout mice. Objective 4 2008 1. Continue establishing adult normal ranges for phytosterol, cholestanol and cholesterol precursor levels using samples collected during Cycle 6 of the Framingham Offspring Study. Objective 3 2. Continue defining the relationship between phytosterol, cholestanol and cholesterol precursor levels and plasma lipid, lipoprotein and apolipoprotein levels. Objective 3 3. Continue defining the relationship between phytosterol, cholestanol and cholesterol precursor levels and selected dietary intake data (macronutrients, cholesterol, fatty acids, fiber and antioxidant supplements). Objective 3 4. Continue determining the effect of dietary fatty acids on mRNA abundance of genes involved with the metabolism of low-density lipoprotein (LDL) cholesterol and high-density lipoprotein cholesterol in Golden Syrian Hamsters. Objective 4 5. Continue determining the effect of dietary fatty acids on mRNA abundance of genes involved with the metabolism of low-density lipoprotein (LDL) cholesterol and high-density lipoprotein cholesterol in LDL knockout mice. Objective 4 2009 1. Complete studies determining adult normal ranges for phytosterol, cholestanol and cholesterol precursor levels using samples collected during Cycle 6 of the Framingham Offspring Study. Objective 3 2. Continue work defining the relationship between phytosterol, cholestanol and cholesterol precursor levels and plasma lipid, lipoprotein and apolipoprotein levels. Objetive 3 3. Continue work defining the relationship between phytosterol, cholestanol and cholesterol precursor levels and selected dietary intake data (macronutrients, cholesterol, fatty acids, fiber and antioxidant supplements). Objective 3 4. Continue work determining the effect of dietary fatty acids on mRNA abundance of genes involved with the metabolism of low-density lipoprotein (LDL) cholesterol and high-density lipoprotein cholesterol in Golden Syrian Hamsters. Objective 4 5. Continue work determining the effect of dietary fatty acids on mRNA abundance of genes involved with the metabolism of low-density lipoprotein (LDL) cholesterol and high-density lipoprotein cholesterol in LDL knockout mice. Objective 4 4a.List the single most significant research accomplishment during FY 2006. Glycemic Load and Weight Loss The Lipid Metabolism Laboratory found that obese, free-living subjects who consumed caloric-restricted, low glycemic index diets experienced greater weight loss, greater reductions in insulin and triglyceride levels, and greater increases in high density lipoprotein cholesterol over a one-year period than those who followed high glycemic index diets. The results of this work are closely aligned to National Program 107 - Human Nutrition program component: 6. Prevention of Obesity and Disease: Relationship between Diet, Genetics, and Lifestyle. 4b.List other significant research accomplishment(s), if any. Docosahexaenoic Acid and Dementia In a 9-year follow-up study of 899 elderly male and female participants in the Framingham Heart Study, scientists in the Lipid Metabolism Laboratory tested the hypothesis that fish intake and level of certain plasma fatty acids found in fish would be associated with a decreased risk of developing dementia. Our data indicated that subjects with increased fish intakes (over 2 servings per week) and elevated blood levels of plasma n3 fatty acids (docosahexaenoic acid) showed an approximate 50 percent reduction in risk of new onset all-cause dementia and Alzheimer’s disease. These data indicate that intake of fish or fish oil may reduce the risk of developing dementia and cognitive dysfunction in the elderly. This finding needs to be tested in a randomized prospective trial. This work is related to National Program 107 - Human Nutrition program component 6. Prevention of Obesity and Disease: Relationship Between Diet, Genetics, and Lifestyle. Palm, partially-hydrogenated soybean oil, soybean oil and canola oil and CVD risk factors The Cardiovascular Nutrition Laboratory completed studies on the effect of palm oil relative to soybean oil, partially hydrogenated soybean oil and canola oil on cardiovascular disease risk factors in middle-aged and older hypercholesterolemic female and male subjects.The problem addressed is whether palm oil is an acceptable substitute for partially-hydrogenated soybean oil as assessed on the basis of plasma lipoprotein levels and other heart disease risk factors. The effects of these two fats were assessed relative to soybean oil, rich in polyunsaturated fatty acids and canola oil, rich in monounsaturated fatty acids. Fifteen subjects over the age of 50 years with low density lipoprotein (LDL) cholesterol concentrations greater than 130 mg/dL at screening were fed each of five experimental diets provided in random order for 35 day periods each (partially-hydrogenated soybean, soybean, palm, or canola; 2/3 fat, 20% energy). Plasma fatty acid profiles, lipids, lipoproteins, apo AI, apo B, Lp(a), glucose, insulin, HDL subfractions, and indicators of lipoprotein metabolism (HDL cholesterol fractional esterification rate, and CETP, PLTP and paraoxonase activities) were measured at the end of each phase. Plasma fatty acid profiles reflected the major source of dietary fat. Partially-hydrogenated soybean and palm oils resulted in higher LDL cholesterol and apo B concentrations than soybean and canola. There was no significant effect of palm oil relative to the other dietary fats on the total cholesterol/HDL ratio. HDL3 cholesterol was higher after palm oil relative to partially-hydrogenated and soybean oils. Differences attributable to dietary fat in measures of glucose and HDL intravascular processing were small. Palm and partially-hydrogenated soybean oils, relative to soybean and canola oils, adversely altered the lipoprotein profile in moderately hyperlipidemic subjects without significantly affecting HDL intravascular processing markers. This work is related to National Program 107 - Human Nutrition program component 6. Prevention of Obesity and Disease: Relationship Between Diet, Genetics, and Lifestyle. 4c.List significant activities that support special target populations. None. 4d.Progress report. None. 5.Describe the major accomplishments to date and their predicted or actual impact. The first major accomplishment of the Lipid Metabolism Laboratory over the life of this project our finding that the weight loss associated with following popular diets in overweight and obese subjects in the free-living state is not a function of carbohydrate restriction (Atkins diet), fat restriction (Ornish diet), or decreased glycemic load or increased protein (Zone diet), but rather a function of caloric restriction (Weight Watchers program). Compliance to any given dietary program that promotes lowering of caloric intake can result in significance weight loss. (Objective 1, Milestone 1, 2005). The second major accomplishment is the finding among Framingham Heart Study subjects that decreased levels of large alpha-1 migrating HDL particles in plasma as determined by two dimensional gel electrophoresis is a much better predictor of heart disease than is low HDL cholesterol. This finding has important implications in the assessment of heart disease risk. (Objective 5, Milestone 2, 2006). This fraction of HDL is very effective in promoting delivery of cholesterol to the liver via the SRB1 receptor, while very small pre-beta migrating HDL serve as optimal acceptors of free cholesterol from cells via the ABCA1 transporter. These data explain the exact functions of these particles an illustrate of reverse cholesterol transport and heart disease risk from imaging HDL particles in whole plasma. The technology provides a very precise measure of heart disease risk and allows us to examine the effects of individual interventions on these particles, in predicting the potential utility of such dietary interventions in reducing heart disease risk. Free-living, obese subjects consuming caloric restricted, low glycemic index diets experienced greater weight loss, greater reduction in plasma insulin and triglyceride levels, and greater increase in high density lipoprotein cholesterol over a one-year period than those who followied high glycemic index diets. These findings have important implications for developing optimal diets for prevention of obesity, diabetes, and heart disease (Objective 2, Milestone 1, 2006). The fourth major accomplishment is that elevated plasma phospholipid docosahexaenoic acid (DHA) levels that are associated with increased fish intake of more than two servings per week reduce the risk of all-cause dementia and Alzheimer’s disease by about 50 percent. These findings have important implications for the development of nutritional strategies for the prevention of dementia in the elderly. The further accomplishments of the Laboratory relate to the following components of the National Program 107 – Human Nutrition: 4 - Nutrient requirements, 5 - Health promoting properties of plant and animal foods, 6 - Prevention of obesity and disease: relationship between diet, genetics, and lifestyle, and 7 - Health promotion intervention strategies in target populations. The first major accomplishment of the Cardiovascular Nutrition Laboratory over the life of the project includes work related to Milestone 4: Cholesterol homeostasis. In a case-controlled study of Framingham Offspring Study participants with coronary heart disease and carotid stenosis, cardiovascular health was associated with lower absorption rates and paradoxically higher synthesis rates of cholesterol. These surrogate markers of cardiovascular health were better predictors of risk than plasma lipid levels. These data provide support for using surrogate markers of cholesterol homeostasis as risk factor measures, in addition to those currently in place (i.e., plasma lipids, c-reactive protein [CRP] levels) to assess an individual’s risk for cardiovascular health and will promote cost effective and targeted diagnostic and treatment modalities. Corn oil (low in trans fatty acid) is currently being use in the commercial preparation of savory snacks as a replacement for hydrogenated fat (high trans fatty acids). The impact of these two oils was assessed in moderately hypercholesterolemic postmenopausal females and older males. The fats were tested under conditions where they contributed a relatively high amount (20%) of the dietary fat and were incorporated into various food items throughout the day. Scientists observed a significant decrease in total and LDL cholesterol levels, in both the fasting and non-fasting states among subjects consuming diets prepared with the test corn oil relative to the hydrogenated fat. These results suggest that encouraging the food industry to displace trans fatty acid-containing hydrogenated fats with fats low in trans fatty acids may have a beneficial effect with respect to cardiovascular health. This work was related to Milestone 1: Dietary variables that alter cardiovascular disease risk. Metabolic study of selectively bred and genetically modified oils. These accomplishments will have the following potential benefits by providing a basis with which to formulate more efficacious public health dietary guidelines to maintain cardiovascular health for both the general population and high-risk individuals. They can also be used to design more accurate tools for identifying individuals at increased risk of developing cardiovascular disease at an early age, thereby fostering an economically sound model of prevention and maintenance of cardiovascular health rather than treatment of disease. This information can be used to relate cholesterol homeostasis regulation to dietary intake data and specific genotypes associated with lipoprotein metabolism to provide a better understanding of the environmental and genetic influences determining cardiovascular health, and to increase our understanding at a basic level of how specific dietary modifications alter the metabolic parameters that favor cardiovascular health at the genetic level. These accomplishments relate to the National Program 107 - Human Nutrition component 6 - Prevention of Obesity and Disease: Relationship between Diet, Genetics. 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? Two methodologies developed in the Lipid Metabolism Laboratory over the past year that have been transferred to other laboratories. These methods include: two dimensional gel electrophoresis for the analysis of HDL subfractions from whole plasma or serum; and uman apolipoprotein metabolic study methodology with stable isotopes in the fed tate to examine endogenous labeling of apolipoproteins and their metabolism. These indings have been presented at national meetings and have been reported in peer-reviewed journals. There are no constraints on the use of this technology. Our research findings have been published in peer-reviewed journals as included in this report. Research accomplishments have also been presented to the scientific community at a number of meetings including the American Heart Association’s symposium on Obesity, Lifestyle and Lifestyle, the 2006 Women’s Cardiac Health Conference of the Massachusetts Medical Society, the Nutrition and Health: State of the Science & Clinical Applications at Columbia University, and the American Diabetes Association’s annual meeting. 7.List your most important publications in the popular press and presentations to organizations and articles written about your work. (NOTE: List your peer reviewed publications below). Lipid Metabolism Laboratory Articles and Presentations: 1. Article in USDA Bulletin, May 2006 about our Popular Diet Study 2. Cardiology Grand Rounds, Massachusetts General Hospital, Boston, MA. Invited speaker. Where are we with HDL Raising? January 2006. 3. Cardiology Grand Rounds, University of Chicago Hospital, Chicago, IL. Invited speaker. Lipoprotein (a) and Premature Heart Disease, March 2006. 4. Boston University School of Medicine, invited speaker. Benefits of n3 Fatty Acids in Prevention of Heart Disease and Dementia, May 2006. 5. International Symposium of Atherosclerosis, Rome. Keynote speaker. Diet and Heart Disease: Where Are We? June 2006. Also speaker at satellite meetings in Naples and Parma. 6. Endocrine Society Meetings, Boston, MA. Advances in the Diagnosis and Treatment of Lipid Disorders, June 2006. Cardiovascular Research Laboratory Articles and Presentations: 1. Chicago Daily Tribune, October 11, 2005, By Ronald Kotulak, Eating fish slows loss of memory, study says 2. The Miami Herald, October 16, 2005, By Amy Driscoll, Trim calories and pump up nutrition-packed fruits and veggies to help hold back the hands of time 3. The Boston Globe, October 24, 2005, By Beth Daley, Fish warnings taken too far 4. The New York Times, October 31, 2005, By Alexei Barrionuevo, An Apple a Day for Health? Mars Recommends Two Bars of Chocolate 5. Real Simple, October 2005, By Mary Desmond Pinkowish, Check the label 6. Parade Magazine, November 13, 2005, Victuals vs. Vitamins 7. MedPage Today, November 17, 2005, By Peggy Peck, AHA: Fish Oil Plus Statin Reduces Coronary Events 8. MedPage Today, November 22, 2005, By Katrina Woznicki, Chocolate Makers Fortify Cocoa Snacks for Health 9. CBS4 News, November 22, 2005, Reporter Paula Ebben, New Food Labels Help ID Carbs, Whole Grains 10. CBS4 Boston, Nov 23, 2005, Reporter Dr. Mallika Marshall, Can It Be True? Chocolate That's Good For You? 11. Nutrition Action Health Letter, November 2005, By Bonnie Liebman, Cover Story: Antioxidants: Still Hazy After All These Years 12. Nutrition Action Health Letter, November 2005, By Bonnie Liebman, Cover Story: Beyond Vitamins 13. CBS13, CA, December 1, 2005, Healthcheck: Chocolate Bar Good For Your Heart? 14. Toledo Blade, December 5, 2005, By Luke Shockman, Trans fat: 'Zero' foods add up: Experts say manufacturers should have to list even traces of the substance 15. MedPage Today, December 7, 2005, By Katrina Woznicki, IOM Report Says TV Advertisers Influences Children's Eating Habits 16. WCCO, MN, December 8, 2005, Reporter Dennis Douda, 'Healthy' Chocolate Bar Could Tax Your Diet 17. WFRV, CBS 5, Boston, December 8, 2005, Reporter Lisa Malak 18. National Public Radio, December 15, 2005, Host Renee Montagne 19. TheBostonChannel.com, December 22, 2005, New Food Label Requirements To Go Into Effect: Trans Fat Content To Be Added To Labels 20. U.S. News & World Report, December 26, 2005, By Kerry Hannon, Have a Daily Dose of Omega-3: From food to floss: how to stay in good shape in 2006 and beyond 21. The New York Times, December 28, 2005, By Melanie Warner, Marketing Fortified Food to Those Leery of Drugs 22. HealthDay via Gannett News Service, December 30, 2005, By Janice Billingsley, Trans fat food labels are coming 23. SeaFood Business, December 2005, By Steven Hedlund, Medical miracle: omega-3 24. WebMD, January 6, 2006, By Kathleen Zelman 25. Medpage Today, January 13, 2006, By Katrina Woznicki, Fat Viewed by Americans as Less of Stigma 26. Newsweek, January 16, 2006, By Anne Underwood, The Nature of Nutrients: Are you really getting all the vitamins, minerals and fatty acids you need? 27. The Los Angeles Times, January 23, 2006, By Rosie Mestel, Zero guilt? Not on your life. 'No trans fat' doesn't mean low-fat. It might even mean more fat. 28. USA Today, January 23, 2006, By Kim Painter, Today's mailbag 29. National Public Radio, January 24, 2006, Reporter Allison Aubrey, Host Renee Montagne, Study: Fish Oil May Not Help Prevent Cancer 30. HealthScout News (South Africa), January 30, 2006, Don't skip breakfast 31. Medwise News / Fashion Monitor (Toronto Canada), January 31, 2006, Cholesterol Lowering Margarines Don't Promise Miracles 32. International Food Information Council Food Insight Newsletter, January/February 2006, Make the Most Out of Dietary Fats Information: Keep the Trans AND Saturated Fats Down 33. The New York Times, February 10, 2006, To the Editor: Re "Low-Fat Diets Flub a Test" (editorial, Feb. 9): 34. The News Journal (Delaware), February 14, 2006, By In-Sung Yoo, Snacks claim health benefits of chocolate 35. The Washington Post, February 14, 2006, By Sally Squires, Chewing the Low-Fat Results 36. The News-Times, February 16, 2006, By Robert Miller, Area doctors still stick to low-fat diet message 37. WebMD, February 17, 2006, By R. Morgan Griffin, The New Low-Cholesterol Diet: Soy 38. WebMD, February 22, 2006, By Salynn Boyles, Low-Salt Diet May Not Cut Heart Risk: More Deaths Seen With Sodium Restriction 39. USA Today, February 27, 2006, By Elizabeth Weise, Pattern for health is tried and true: Amid conflicting diet headlines, experts advise staying the course 40. Bottom Line's Daily Health News, February 28, 2006, SPILLING THE BEANS ON BEANS 41. O, The Oprah Magazine, February 2006, By Alison Motluk, The Drug Detectives: Women who make sure your medication is safe 42. Eating Well, February 2006, By Peter Jaret, The Food-Cancer Connection 43. Delicious Living, February 2006, By Cheryl Sternman Rule, How diet can help you heal after a heart attack or cardiac surgery 44. Harvard Health Letter, February 2006, Take two candy bars and call me in the morning? Adding plant sterols and flavanols to chocolate doesn't make candy bars a health food. 45. Doc News, March 1, 2006, By Joene Hendry , Healthful Fiber Findings Often Forgotten: Benefits include lowering CVD risk, normalizing glucose 46. The Los Angeles Times, March 6, 2006, By Hilary E. MacGregor, Chocolate, with benefits? Forget the nuts and caramel. Candy makers are packing sweets with heart-healthy ingredients 47. WebMD, March 7, 2006, By Louise Chang, Coffee May Up Heart Risks for Some 48. U.S. News & World Report, March 8, 2006, By Christopher J. Gearon, Retirement: Five tips for the next 20 years 49. Tribune Media Services, March 8, 2006, By Bev Bennett, Nutritious fish on a fast track 50. Scout News Service, March 12, 2006, By Steven Reinberg, Folic Acid Supplements Won't Lower Heart-Attack Risk 51. Associated Press, March 15, 2006, Tofu, oatmeal can help lower cholesterol: Study finds strict soy, nut, fiber diet can be as effective as drugs 52. Scout News Service, March 16, 2006, By Kathleen Doheny, Foods Fortified With Folic Acid May Cut Stroke Risk 53. The Washington Post, March 21, 2006, By Sally Squires, Portfolio Diet: Tough to Stay Invested In 54. The Los Angeles Times, March 27, 2006, By Denise Gellene, Makin' better bacon: Scientists use genetic engineeringto make pigs rich in healthy fat 55. The New York Times, March 27, 2006, By Gina Kolata 56. The New York Times, March 28, 2006, Corrections: For the Record 57. Orlando Sentinel, March 28, 2006, By Robyn Shelton, Anti-cholesterol diet unlikely to spark fad 58. Harvard Heart Letter, March 2006, Eating your way to lower cholesterol 59. Science News, April 1, 2006, By Christen Brownlee, Pigging Out Healthfully: Engineered pork has more omega-3s 60. Prevention.com, April 11, 2006, Oh No, It's Frankenfat! 61. Associated Press, April 18, 2006, By Jamie Stengle, A yo-yo sized lesson in portion control 62. Health Day News, April 24, 2006, By Ed Edelson, Heavy Coffee Drinking Doesn't Hurt the Heart: No risk of heart disease found in two long-term studies 63. The Los Angeles Times, May 8, 2006, By Hilary E. MacGregor, Fab bean or has-been? 64. Chicago Tribune, May 17, 2006, By Janet Helm, When fats are good; Time to rethink the low-fat mantra in favor of healthful oils and fats 65. USA Today (Weekend magazine), May 21, 2006, By Jean Carper, A guide to better health with the AHA 66. Shape Magazine, May 2006, By Connie Harikul, The latest on what makes a diet healthy 67. O, The Oprah Magazine, May 2006, By Allison Motluck, More Omega-3s, Please 68. Women’s Health, June 6, 2006, By Lisa Draver, Food labels can stump even the smartest eater. Here's our cheat sheet for the back of the box 69. Tribune Media Services, June 8, 2006, By Bev Bennett, Fish is a Fast Meal, and It’s Good for You 70. American Public Radio- Marketplace, June 8, 2006, Host Helen Palmer, Wendy's gets leaner 71. MSN Health and Fitness, June 8, 2006, By Rich Maloof, Reality Check Health Centers: Cholesterol-High cholesterol is within your power to control. 72. Health Day News, June 13, 2006, By Ed Edelson, Fish Oil Won't Fix Abnormal Heart Rhythms: Study 73. Fitcommerce.com, June 14, 2006, Hear Me Roar, Time For Change -- Understanding Women's Heart Disease 74. Eating Well, June/July 2006, By Peter Jaret, Soy Reconsidered; Is the magic bean a magic bullet? 75. The Chicago Tribune, June 18, 2006, Extra-crispy chicken and deep-fried panic 76. WebMD, June 19, 2006, By Daniel DeNoon, Lifestyle Vital to New Heart Diet: American Heart Association: Good Food, Exercise Best Way to Health 77. NBC Newsfeed, June 19, 2006, HealthWatch 78. Health Day News, June 19, 2006, By Kathleen Doheny, Veggies Do a Heart Good: Mice fed 30% vegetable diet were thinner with clearer arteries, study found 79. Heartwire, June 19, 2006, New AHA diet recommendations also emphasize lifestyle, but focus is still mostly on trimming the fat 80. Investor’s Business Daily, June 19, 2006, Issues & Insights: Let Them Eat Whatever 81. MedPage Today, June 20, 2006, By Peggy Peck, AHA Recommends Less Fat and Sugar and More Exercise 82. Associated Press, June 20, 2006, By Marilynn Marchione, Guidelines call for limits on trans fats: The American Heart Association lends its support to a dietary crackdown. 83. The Detroit News, June 20, 2006, By Kimberly Hayes Taylor, The ABCs of the vitamin debate: Should you take dietary supplements? Physicians disagree over conflicting studies.