Location: Diet, Genomics and Immunology Laboratory2019 Annual Report
Objective 1: Determine the accumulation and variability of important bioactive compounds in commonly consumed food crops as affected by factors such as stage of maturity (leafy greens harvested at various stages of maturity) and variety/processing (coffee products derived from different sources). [NP107, C1, PS1A] Objective 2: Determine bioavailability and cellular uptake of potential compounds in coffee products; investigate their effects on subclinical inflammation and its associated events related to chronic metabolic diseases; elucidate mechanisms of action on subclinical inflammation and related events. [NP107, C3, PS3B] Objective 3: Determine effects of brassica vegetables harvested at different stages of maturity (e.g., sprout, microgreen, baby green, mature plant) on high fat diet-induced inflammation, and adipose uncoupling protein 1 (UCP1) as mechanisms for their attenuation of high fat diet-induced weight gain. Elucidate the role of the microbiome in mediating changes in inflammation and liver lipid metabolism. [NP107, C3, PS3B]
Objective 1: Amounts of bioactive compounds in selected model plant products (e.g., coffee beans grown in different regions/conditions, coffee products, brassica vegetables harvested at different stages of maturity) will be determined using established HPLC, MS/MS and NMR methods. Objective 2: Both in-vivo and in-vitro models will be used for this objective. (1) We will determine bioavailability and cellular uptake of coffee compounds using cell culture (e.g., Caco-2, HepG2, monocytic THP-1) models. HPLC, metabolomic and lipidomic analytical technologies will be used to measure the compounds and associated metabolites. (2) We will determine potential effects of coffee/coffee chemicals (javamide-I/-II) on subclinical inflammation markers using a rodent model. Obesity will be induced in animals (e.g., rats) fed a high-fat diet, and the potential effects of coffee and coffee compounds (e.g., javamide-I/-II) on obesity-associated subclinical inflammation and biological changes will be determined to elucidate the effects. (3) We will determine the cellular/molecular mechanisms responsible for the biological effects using cell culture models. Cell will be treated with the compounds and effects of compounds on cellular pathways related to signal transduction pathways, inflammatory cytokines, adhesion molecular, transcriptional factors will be assessed at protein and message levels using western blots, ELISA and RT-PCR. Objective 3: We will determine effects of brassica vegetables harvested at different stages of maturity (e.g., sprout, microgreen, baby green, mature plant) on high fat diet-induced rodent model. Inflammatory marker and adipose uncoupling protein 1 (UCP1) will be assessed using ELISA at protein level and RT-PCR at message level as mechanisms for their attenuation of high fat diet-induced weight gain. Biochemical and marker genes analysis will be performed in liver and adipose tissues to assess the effects on lipid and energy metabolism. Metagenomic analysis using next generation sequencing techology will be performed to elucidate the role of the microbiome in mediating changes in inflammation and liver lipid metabolism.
This report is for a new NP107 OSQR approved project entitled “Elucidating Phytonutrient Bioavailability, Health Promoting Effects and Mechanisms of Existing/Emerging Foods and Beverages.” Several reports suggest that plant-based foods and drinks may have beneficial health effects on several human diseases (e.g., obesity, diabetes, liver disease, cardiovascular disease, cancer) but the bioactive composition as well as mechanisms of action remain largely unclear. The current project seeks to address the deficiencies. Coffee contains several bioactive compounds (e.g., caffeine, chlorogenic acids, javamide-I/-II), and recent reports indicate that javamide-I/-II may have a greater anti-inflammatory activity than caffeine and chlorogenic acids. However, there is little information about the amounts of javamide-I/-II in different coffee beans and products, so it is unfeasible to assess their potential effects. Furthermore, there is little information regarding the potential impacts of species and growing regions on the amounts of javamide-I/-II in coffee beans. Therefore, a new HPLC method is under development to measure not only javamide-I/-II, but also chlorogenic acids (3-O-caffeoylquinic acid, 4-O-caffeoylquinic acid, 5-O-caffeoylquinic acid) and caffeine concurrently. This new method will make it possible to assess their amounts and biological activities in coffee products related to the proposed studies. Brassica vegetables are also known to contain several bioactives. LC/MS methods were developed for the analysis of glucosinolates and flavonoids in brassica vegetables. It was found that brassica vegetable microgreens in their mature form contain drastically different levels and varieties of glucosinlates and flavonoids. The microgreen/mature vegetable, with composition determined, will be used in the bio-efficacy studies of animal models of chronic disease.