1a.Objectives (from AD-416)
The pilot study will examine the metabolism, safety and efficacy of increased intake of citrus liminoids by human subjects. The main study will determine their health benefits, particularly the effects on serum lipids and markers of inflammation.

1b.Approach (from AD-416)
In the pilot study 6 healthy subjects (18-65y) with LDL cholesterol greater than 130 mg/dL will be recruited and fed citrus liminoids. The metabolites of limonoids in the serum and urine samples will be determined by the use of mass spectrometry. NMR technology will be used to determine the number and sizes and of different lipoprotein fractions in plasma. Markers of inflammation in the serum and also their ex-vivo production after stimulation of blood cells with mitogens will be determined by ELISA and flowcytometry methods. The design, number of subjects, and the dose of liminoids to be used will depend upon the results obtained from the pilot study. Documents Trust with Coca Cola Institute for Health and Wellness. Log 27838. Associated with WRRC A. Breksa project 5325-41430-009-01T 411498 425 Log 32687.

3.Progress Report
This report serves to document research conducted under a trust agreement between ARS and the Coca Cola Beverage Company. Additional details of this research project can be found in the report for the parent CRIS 5306-51530-015-00D, Influence of Dietary Fatty Acids on Human Health with Emphasis on Immune and Inflammatory Responses. We had intended to start our citrus limonoids study in hypercholesterolemic men (not listed in this CRIS), but because of the delay in the purification of the limonoids, we were not able to start it during last year. We held several meetings and conference calls with our sponsors (Beverage Institute for Health and Wellness) and our collaborators (Dr Breksa at WRRC) to solve the problems. We now have enough material for 10 subjects and are currently recruiting subjects for this study. We are also in preparation of transgenic mice that overexpress TLR4 in breast, adipose or prostate tissues. If these mice show enhanced inflammation, these mice will be used to test anti-inflammatory and chemopreventive efficacy of plant polyphenols