Research Project: Elucidating Phytonutrient Bioavailability, Health Promoting Effects and Mechanisms of Existing/Emerging Foods and Beverages

Location: Diet, Genomics and Immunology Laboratory

2023 Annual Report

Objectives
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]

Approach
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 technology will be performed to elucidate the role of the microbiome in mediating changes in inflammation and liver lipid metabolism.

Progress Report
This report is for a NP107 project entitled "Elucidating Phytonutrient Bioavailability, Health Promoting Effects and Mechanisms of Existing/Emerging Foods and Beverages." We focus on two foods, coffee and kale at different growth/maturation states. The following describes the current progress of the project for FY23. For Objective 1, we are assessing the amounts of coffee purported health promoting bioactives javamide-I/-II in decaf and other coffee products in the market using a high performance liquid chromatography (HPLC) method. Also, using a high-performance chemical isotope-labelling LC/MS method, we are in the process of identifying bio-active compounds beyond javamide-I/-II, chlorogenic acids, in coffee products. Preliminary analysis suggests significant disparities in the amounts of javamide-I/-II and other compounds in the coffee products including decaf products and that every coffee product in the market may not provide the same purported health effects. For Objective 2, we are investigating potential effects of coffee containing javamide-I/-II (CCJ12) on metabolic/inflammatory factors in an obese model. We are analyzing metabolic factors (bodyweight, cholesterol, HDL/LDL, lipids), inflammatory cytokines (e.g., TNF-alpha, IL-1beta, IL-6, MCP-1), and adipokines (adiponectin) in plasma samples from the study. We are also investigating the effects of caffedymine-type phenolic compounds found in coffee on signal transduction pathways (e.g., Mitogen-activated protein [MAP] Kinase, NF-KB) and Cyclooxygenase (COX) enzymes. For Objective 3, we are examining the effects of kale (Darkibor variety) microgreen and its mature counterpart on the gut microbiome. We are employing a mouse model of diet-induced obesity for this work. Animals (C57BL mice) are being fed with diets contain low fat (10%, LF)) or high fat (45%, HF), supplemented with or without Kale microgreen (KMG) or mature kale (MK) in the perspective diet matrix for 8 weeks. We are processing data to elucidate potential biomarkers that may correlated with KMG and MK consumption in both low fat and high matrix. Modulatory effects of consuming KMG, MK on the gut microbiome is being assessed; and influence of diet matrix are also being examined. Preliminary analysis identified an unidentified Ruminococcus species, as a potential biomarker to be correlated with KMG and MK consumption in both low fat and high matrix.

Accomplishments
1. Individual differences in biology and inflammatory responses of adipose stem cell identified. Adipose stem cell plays critical roles in normal physiology as well as diseases development. Understanding individual differences in adipose stems cell biology and responses to stimuli serve as critical step toward realizing the goal of precision nutrition. ARS scientists in Beltsville, Maryland, examines the growth, differentiation, and responses of adipose stem cell to inflammatory stimuli in adipose stem cells from different individuals. Individual differences in growth, differentiation and inflammatory responses were identified. The study provides fundamental information regarding individual difference needed for precision nutrition.

Review Publications
Schillinger, D., Bullock, A., Powell, C., Fukagawa, N.K., Greenlee, C., Towne, J., Gonzalvo, J.D., Lopata, A.M., Cook Iv, W.J., Herman, W.H. 2023. Leveraging federal programs for population-level diabetes prevention and control: Recommendations from the National Clinical Care Commission. Diabetes Care. 46(2):e24-e38. https://doi.org/10.2337/dc22-0619.
Boltri, J., Tracer, H., Strogatz, D., Schumacher, P., Fukagawa, N.K., Shell, D., Wu, S., Idzik, S., Herman, W., Powell, C. 2023. The NCCC Report to Congress: Prevention in people with prediabetes. Diabetes Care. 46(2):e39-e50. https://doi.org/10.2337/dc22-0620.
Schillinger, D., Bullock, A., Powell, C., Fukagawa, N.K., Greenlee, C.M., Towne, J., Gonzalvo, J.D., Lopata, A.M., Cook IV, W., Herman, W.H., Albright, A., Bolen, S., Boltri, J.M., Chong, W., Conlin, P.R., Dokun, A., Hawkins, M., Idzik, S., Leake, E., Linder, B., Marx, B., Schumacher, P., Shell, D., Strogatz, D., Tracer, H., Wu, S. 2023. The National Clinical Care Commission Report to Congress: Recommendations to better leverage federal policies and programs to prevent and control diabetes. Diabetes Care. 46(2):255-261. https://doi.org/10.2337/dc22-1587.
Liu, F., Smith, A.D., Wang, T.T., Pham, Q., Yang, H., Li, R.W. 2023. Multi-omics analysis detected multiple pathways by which pomegranate punicalagin exerts its biological effects in modulating host–microbiota interactions in murine colitis models. Food & Function. 14:3824. https://doi.org/10.1039/D3FO00286A.
Liu, F., Smith, A.D., Wang, T.T., Pham, Q., Yang, H., Li, R.W. 2023. Ellagitannin punicalagin disrupts the pathways related to bacterial growth and affects multiple pattern recognition receptor signaling by acting as a selective histone deacetylase inhibitor. Journal of Agricultural and Food Chemistry. 71(12):5016-5026. https://doi.org/10.1021/acs.jafc.2c08738.
Liu, F., Smith, A.D., Wang, T.T., Pham, Q., Cheung, L., Yang, H., Li, R.W. 2022. Biological pathways via which the anthocyanin malvidin alleviated the murine colitis induced by Citrobacter rodentium. Food & Function. 14(2):1048-1061. https://doi.org/10.1039/D2FO02873E.
Yasmin, R., Pham, Q., Fukagawa, N.K., Wang, T.T. 2022. Individual variabilities in adipose stem cell proliferation, gene expression and responses to lipopolysaccharide stimulation. International Journal of Molecular Sciences. 23:12534. https://doi.org/10.3390/ijms232012534.
Li, Y., Liu, Z., Tamia, G., He, X., Sun, J., Chen, P., Lee, S., Wang, T.T., Gao, B., Xie, Z., Yu, L. 2022. Identification of soluble free, soluble conjugated and insoluble bound phenolics in tomato seeds and their radical scavenging and antiproliferative activities. Journal of Agricultural and Food Chemistry. 70(29):9039-9047. https://pubs.acs.org/doi/10.1021/acs.jafc.2c03418.
Xie, Z., Li, Y., Liu, Z., Zeng, M., Moore, J., Gao, B., Wu, X., Sun, J., Wang, T.T., Pehrsson, P.R., He, X., Yu, L. 2023. Chemical characterization of cinnamon (Cinnamomum verum J. Presl) extracts and their capacities in weakening SARS-CoV-2 spike protein binding to ACE2, inhibiting ACE2 and scavenging free radicals. Journal of Agricultural and Food Chemistry. 71:4890-4900. https://doi.org/10.1021/acs.jafc.3c00285.
Park, J.B., Peters, R.C., Novotny Dura, J. 2023. Impact of roasting on javamide-I/-II in Arabica and Robusta coffee beans. Food Chemistry. 412:135586. https://doi.org//10.1016/j.foodchem.2023.135586.