Progress 11/15/15 to 11/14/21
Outputs
Target Audience:The target audience for identification of biomarkers for rice bran and legumes wereChildren and Adults residing in the US that are at high risk for developing chronic diseases. These diseases included, but were not limited to obesity, diabetes, cardiovascular disease and cancer. Changes/Problems:There were delays in final years of the project with COVID-19 lab shutdowns. These delays limited thefinalvalidation of targeted quantification approaches of biomaker metabolites. What opportunities for training and professional development has the project provided?Multidisciplinary cross training between technical staff, undergraduate and graduate students from colleges of agriculture, food science, toxicology/environmental healthand biomedical sciences. How have the results been disseminated to communities of interest?Manuscripts are under reviewfor publication in peer reviewed journals. Presentations were conducted at the North America Society of Metabolomics and at the American Society of Nutrition. What do you plan to do during the next reporting period to accomplish the goals?
Nothing Reported
Impacts
What was accomplished under these goals?
Non-targeted and targeted metabolomics identifies dietary exposure biomarkers for navy bean and rice bran consumption in children and adults: a secondary analysis from randomized control trials Background: Traditional methods for assessing dietary exposures that rely on self-reporting are 2 error prone. Qualitative and quantitative small molecule markers following food consumption are 3 needed for establishing relationships with health and disease outcomes. 4 Objective: The main objective was to examine dietary exposure biomarkers for navy beans and 5 rice bran using targeted and non-targeted metabolomics. 6 Methods: Plasma, urine, stool, and study foods from three randomized, controlled, dietary 7 intervention studies were subjected to secondary analysis by ultra-performance liquid-8 chromatography tandem mass-spectrometry (UPLC-MS/MS). Cohort 1 (N = 38 children), cohort 9 2 (N=29 adults) and 3 (N = 20 adults) consuming navy beans and/or rice bran for 2-, 4-, 6- or 12-10 weeks. 11 Results: Non-targeted metabolomics identified 138 and 175 metabolites of relevance to navy bean 12 and/or rice bran exposure in the diet. Trigonelline concentrations was increased in children (p-13 value 0.04, and 004) and adult (p-value 0.05, and 0.03) plasma after 4 weeks from navy bean or 14 rice bran intake compared to baseline. Plasma pipecolate concentrations after 4 weeks of navy 15 bean intake increased in both children and adults and decreased in adult urine. An increase in 16 xanthurenate (46%) was observed in the children plasma after rice bran intake for four weeks. 17 Adult urine S-methylcysteine sulfoxide showed 50% increase after at 2 weeks, and 40% after 4 18 weeks after navy bean intake. Adult urine and stool salicylate decreased (40%) when compared to 19 control after consuming rice bran. Navy bean + rice bran consumed together in adults resulted in 20 higher concentrations of plasma pipecolate (286%) and S-methylcysteine (65%). 21 3 Conclusion: Plasma trigonelline was identified as a candidate dietary biomarker for adults 22 consuming rice bran (30g/day) or navy bean (35g/day) and in children (15g/day) after 2 and/or 4 23 weeks. Non-targeted metabolomics was an effective screening tool for identification of dietary 24 exposure biomarkers. 25 Clinical trial registry numbers: Healthy Hearts clinicaltrails.gov ( clinical trail #NCT01911390). 26 Beans/Bran Enriching Nutritional Eating For Intestinal health Trial (BENEFIT) (clinical trial # 27 NCT01929122).
Publications
- Type:
Journal Articles
Status:
Accepted
Year Published:
2021
Citation:
Zarei I, Baxter BA, Oppel RC, Borresen EC, Brown RJ, Ryan EP*. Plasma and Urine Metabolite Profiles Impacted by Increased Dietary Navy Bean Intake in Colorectal Cancer Survivors: A Randomized-Controlled Trial. Cancer Prevention Research (Phila). 2021 Apr;14(4):497-508. doi: 10.1158/1940-6207.CAPR-20-0270. Epub 2020 Dec 24. PMID: 33361317; PMCID: PMC8026539.
- Type:
Journal Articles
Status:
Accepted
Year Published:
2019
Citation:
Zarei I, Oppel RC, Borresen EC, Brown RJ, Ryan EP* (2019) Modulation of plasma and urine metabolome in colorectal cancer survivors consuming rice bran. Integr Food Nutr Metab 6: DOI: 10.15761/IFNM.1000252
- Type:
Journal Articles
Status:
Under Review
Year Published:
2022
Citation:
Bridget A. Baxter1, Katherine J. Li1, Iman Zarei1, Linxing Yao2, Sangeeta Rao3, Elizabeth P. Ryan1*
Non-targeted and targeted metabolomics identifies dietary exposure biomarkers for navy bean and rice bran consumption in children and adults: a secondary analysis from randomized control trials
- Type:
Journal Articles
Status:
Accepted
Year Published:
2022
Citation:
Brooke Sayre-Chavez, Bridget Baxter, Corey D. Broeckling, Mar�a Mu�oz-Amatria�n, Mark Manary, Elizabeth P. Ryan*. Non-targeted metabolomics of cooked cowpea (Vigna unguiculata) and pigeon pea (Cajanus cajan) from Ghana using two distinct and complementary analytical platforms. Food Chemistry: Molecular Sciences (2022, in press)
- Type:
Journal Articles
Status:
Accepted
Year Published:
2022
Citation:
Pfluger BA, Smith HV, Weber AM, Ibrahim H, Doumbia L, Bore A, Cissoko A, Douyon S, Kone K, Sangare L, Maiga A, Koita OA, Goodman K, Evans AM, Ryan EP*. Non-Targeted Dried Blood Spot-Based Metabolomics Analysis Showed Rice Bran Supplementation Effects Multiple Metabolic Pathways during Infant Weaning and Growth in Mali. Pediatric Nutrition section. Nutrients. 2022; 14(3):609. https://doi.org/10.3390/nu14030609
- Type:
Journal Articles
Status:
Accepted
Year Published:
2021
Citation:
7. Annika M. Weber, Bridget A. Baxter, Anna McClung, Molly M. Lamb, Sylvia Becker-Dreps, Samuel Vilchez, Ousmane Koita, Frank Wieringa, Elizabeth P. Ryan*. Arsenic speciation in rice bran: Agronomic practices, postharvest fermentation, and human health risk assessment across the lifespan. Environmental Pollution, Volume 290,2021,https://doi.org/10.1016/j.envpol.2021.117962.
- Type:
Journal Articles
Status:
Accepted
Year Published:
2021
Citation:
11. Santos MCB, Barouh N, Durand E, Bar�a B, Robert M, Micard V, Lullien-Pellerin V, Villeneuve P, Cameron LC, Ryan EP, Ferreira MSL, Bourlieu-Lacanal C. Metabolomics of Pigmented Rice Coproducts Applying Conventional or Deep Eutectic Extraction Solvents Reveal a Potential Antioxidant Source for Human Nutrition. Metabolites. 2021 Feb 15;11(2):110. doi: 10.3390/metabo11020110. PMID: 33671946; PMCID: PMC7919034.
- Type:
Conference Papers and Presentations
Status:
Accepted
Year Published:
2020
Citation:
15. Elizabeth Ryan*, Bridget Baxter, Katherine Li, Lisa Wolfe, Linxing Yao, Corey Broecling, Erica
Borreson, Lei Zhang, Iman Zarei, Melanie Beale, Sangeeta Rao, Hillary Smith, Luis Zambrana, Ousmane
Koita, and Samuel Vilchez. Developing Biomarkers of Rice Bran and Navy Bean Intake via Integrated
Metabolomics from Infants, Children and Adults for Association with Gut Health Properties, Current Developments in Nutrition, Volume 4, Issue Supplement_2, June 2020, https://doi.org/10.1093/cdn/nzaa045_096
- Type:
Conference Papers and Presentations
Status:
Accepted
Year Published:
2022
Citation:
Madison Tipton, Bridget Baxter, Kailey Berry, Brooke Sayre-Chavez, Mar�a Mu�oz-Amatria�n, Corey D. Broeckling, Mark Manary and Elizabeth P. Ryan. Dried Blood Spots and Urine Metabolite Profiling reveals dietary biomarkers of increased cowpea consumption by young Children and Pregnant Women in Ghana. College of Veterinary Medicine and Biomedical
- Type:
Conference Papers and Presentations
Status:
Accepted
Year Published:
2022
Citation:
National
Baxter BA, Beale M, Leach H & Ryan EP. Rice bran and navy bean intake alters the amino acid profile and healthy eating index of adults at risk for colon cancer. American Society Preventive Oncology (ASPO), Tucson AZ. March 13-15, 2022 (Poster)
|
Progress 11/15/19 to 11/14/20
Outputs
Target Audience:
Nothing Reported
Changes/Problems:Challenges were presented due to lab closures with respect to developing targeted metabolite assays for biospecimens. What opportunities for training and professional development has the project provided?Students and staff were able to focus on data analysis and submit manuscripts. How have the results been disseminated to communities of interest?
Nothing Reported
What do you plan to do during the next reporting period to accomplish the goals?Complete manuscripts and disseminate results in scientific meetings, finish/complete project. There is a final manuscript that is still under review after multiple submissions. Overall the study team has been cited/referenced and established leadership roles in the field for assessing and identifying dietary biomarkers of whole grains and legumes.
Impacts
What was accomplished under these goals?
A no cost extension to finish manuscripts and final analysis of samples was performed while labs were not in full capacity.
Publications
|
Progress 11/15/18 to 11/14/19
Outputs
Target Audience:
Nothing Reported
Changes/Problems:Ryan did have a sabbatical during the fall of 2019 that delayed some onsite development of assays on the mass spectrometers at CSU. What opportunities for training and professional development has the project provided?Networking at meetings. Notably, all of the students have recieved jobs continuing in the field of metabolomics and food science, nutrition with increased education regarding the human health benefits of rice bran and beans across the lifespan. How have the results been disseminated to communities of interest?The university of colorado health collaborating partners have recieved co-authorship and will use findings in nutrition education sessions targeted for K-12, college adults and cancer survivors. What do you plan to do during the next reporting period to accomplish the goals?Finish publications and create the targeted metabolite biomarker assays in the mass spec facility following the list that was generatedof dietary biomarkers identified in the non-targeted metabolomics approach.
Impacts
What was accomplished under these goals?
Training of students on metabolite analysis of foods and biological specimens was followed bydissemination of research through presentations at 3 localschools and grocery stores were completed in addition tofood and nutrition scientific meetings.
Publications
- Type:
Journal Articles
Status:
Accepted
Year Published:
2019
Citation:
Jablonski, B.B.; Carolan, M.; Hale, J.; Thilmany McFadden, D.; Love, E.; Christensen, L.; Covey, T.; Bellow, L.; Cleary, R.; David, O.; Jablonski, K.E.; Jones, A.S.; Meiman, P.; Quinn, J.; Ryan, E.P.; Schipanski, M.; Summers, H.; Uchanski, M. Connecting Urban Food Plans to the Countryside: Leveraging Denvers Food Vision to Explore Meaningful RuralUrban Linkages. Sustainability 2019, 11, 2019
- Type:
Journal Articles
Status:
Accepted
Year Published:
2019
Citation:
Zarei I, Oppel RC, Borresen EC, Brown RJ, Ryan EP* (2019) Modulation of plasma and urine metabolome in colorectal cancer survivors consuming rice bran. Integr Food Nutr Metab 6: DOI: 10.15761/IFNM.1000252
- Type:
Journal Articles
Status:
Accepted
Year Published:
2019
Citation:
Luis E. Zambrana, Starin McKeen, Hend Ibrahim, Iman Zarei, Erica C. Borresen, Lassina Doumbia, Abdoulaye Bore, Alima Cissoko, Seydou Douyon, Karim Kone, Johann Perez, Claudia Perez, Ann Hess, Zaid Abdo, Lansana Sangare, Ababacar Maiga, Sylvia Becker-Dreps, Lijuan Yuan, Ousmane Koita, Samuel Vilchez & Elizabeth P. Ryan*. (2019). Rice bran supplementation modulates growth, microbiota and metabolome in weaning infants: a clinical trial in Nicaragua and Mali. Scientific Reports. 9, Article 13919.
- Type:
Journal Articles
Status:
Accepted
Year Published:
2018
Citation:
Li KJ, Jenkins N, Luckasen G, Rao S, Ryan EP* (2018) Plasma metabolomics of children with aberrant serum lipids and inadequate micronutrient intake. PLoS ONE 13(11): e0205899. PMID: 30412600. https://doi.org/10.1371/journal.pone.0205899
|
Progress 11/15/17 to 11/14/18
Outputs
Target Audience:Our findings for dietary modulation of the metabolome were reached by multidisciplinaryaudiences involved in food chemistry and precision nutrition as well as medical communities including those with interests to prevent chronic disease via nutritional interventions. Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided?The graduate students and research staff that were trained during this time period have succeded to advance studies as postdoctoral fellows and obtaining permanent research scientist positions. There were 3 undergraduates provided with opportunities to co-study biomarkers of whole grains and legumes intake. How have the results been disseminated to communities of interest?There have been four presentations at scientific conferences including but not limited to Society of Toxicology, IFT, American Institute for Cancer Research and American Society of PReventive ONcology. In addition the information has been disseminated in peer-reviewed research publications What do you plan to do during the next reporting period to accomplish the goals?We seek to perform targeted validation of the metabolites using quantification methods on QQQ mass spectrometers at the CSU core metabolomics facility and in collaboration with Metabolon.
Impacts
What was accomplished under these goals?
There were two graduate students trained under this period that resulted infour publications that are provided as final products. Li KJ, Borresen EC, Jenkins-Puccetti N, Luckasen G andRyan EP*Navy Bean and Rice Bran Intake Alters the Plasma Metabolome of Children at Risk for Cardiovascular Disease.2018;Frontiers in Nutrition. 4:71. DOI: 10.3389/fnut.2017.00071. PMID: 29404331 Li KJ, Jenkins N, Luckasen G, Rao S,Ryan EP*(2018) Plasma metabolomics of children with aberrant serum lipids and inadequate micronutrient intake.PLoS ONE13(11): e0205899. PMID: 30412600. https://doi.org/10.1371/journal.pone.0205899 Baxter, B., Oppel, R., andRyan EP*. (2018) Navy beans impact the stool metabolome and metabolic pathways forcolon health in cancer survivors. Nutrients. 2018 Dec 22,11(1).pii:E28. PMID: 30583518 Zarei, I.; Luna, E.; Leach, J.E.; McClung, A.; Vilchez, S.; Koita, O.;Ryan, E.P*.Comparative Rice Bran Metabolomics across Diverse Cultivars and Functional Rice Gene-Bran Metabolite Relationships.Metabolites2018,Oct 9;8(4). pii:E63. PMID:30304872
Publications
- Type:
Journal Articles
Status:
Accepted
Year Published:
2018
Citation:
Baxter, B., Oppel, R., and Ryan EP*. (2018) Navy beans impact the stool metabolome and metabolic pathways for colon health in cancer survivors. Nutrients. 2018 Dec 22,11(1).pii:E28. PMID: 30583518
- Type:
Journal Articles
Status:
Accepted
Year Published:
2018
Citation:
32. Zarei, I.; Luna, E.; Leach, J.E.; McClung, A.; Vilchez, S.; Koita, O.; Ryan, E.P*. Comparative Rice Bran Metabolomics across Diverse Cultivars and Functional Rice GeneBran Metabolite Relationships. Metabolites 2018, Oct 9;8(4). pii:E63. PMID:30304872
- Type:
Journal Articles
Status:
Accepted
Year Published:
2018
Citation:
35. Li KJ, Borresen EC, Jenkins-Puccetti N, Luckasen G and Ryan EP* Navy Bean and Rice Bran Intake Alters the Plasma Metabolome of Children at Risk for Cardiovascular Disease. 2018; Frontiers in Nutrition. 4:71. DOI: 10.3389/fnut.2017.00071. PMID: 29404331
- Type:
Journal Articles
Status:
Accepted
Year Published:
2018
Citation:
Li KJ, Jenkins N, Luckasen G, Rao S, Ryan EP* (2018) Plasma metabolomics of children with aberrant serum lipids and inadequate micronutrient intake. PLoS ONE 13(11): e0205899. PMID: 30412600. https://doi.org/10.1371/journal.pone.0205899
|
Progress 11/15/16 to 11/14/17
Outputs
Target Audience:The target audiencesreached during this project reporting period include undergraduate students in a research symposium, graduate students and academic faculty and staff communities in the fields of food science and chronic disease prevention as well as a number of diverse chemical analytical science groups working in targeted and non targeted metabolomics. The food industry sector has also become aware of our technological advancements and analysis capacity to identify dietary biomarkers of intake for both commonly and uncommonly consumed staple foods in people that have been previously undetermined/unknown. While the human populations that these dietary biomarkers are being assessed has had limited inclusion of racial/ethnicminorities, these findings have included children from lowsocioeconomic groups as evidenced by their enrollment in free/reduced lunch programs. There have been a number of effortsthat our team hasdelivered science-based knowledge to people through both formal and informal educational and community based programs. Ryan has formal classroom instruction, large collaborative team laboratory meetings with students across the ranks presenting in oral and poster platforms. This project has yielded two MPH studentpracticum experiences and experiential learning opportunities through multi-purposed and integrated food and nutritional metabolomics data analysis.Ryan is using data and publications from this effort in the development of curriculum andinnovative teaching methodologies that will be applied to workshops and with respect to the emerging gut microbiome community network of scientists. Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided?This project has provided a diverse set of undergrad and graduate students, research scientists and collaborating facultyfrom the disciplines of food science and nutrition, toxicology, analytical chemistry, epidemiology and statistics. The presentations of findings from this research has provided key professional development opportunities for all staff involved through both national and international scientific meetings and via engagement with food industry stakeholders and metabolomics research groups around the country. How have the results been disseminated to communities of interest?Yes thefirst stage of publications are completed (or in final stages of submission) from the dietary intervention studies in adults and children, and specifically for which the integrated global, non-targeted metabolomics analysis was described for the foods (rice bran and navy beans)as well as the human biological matrices that we can detect these foods after consumption. What do you plan to do during the next reporting period to accomplish the goals?The Objective for this next reporting periodwill encompass together all three specific aims. We seek toidentify candidate dietary biomarkers of intake for navy beans and rice bran from non-targeted metabolomics datasets, and quantify levels of these compounds in a targeted assay. The presence of the plant food metabolite (from rice bran or beans) or its modified/intermediate form (detected in plasma, urine, and/or stool) was confirmed. Food metabolome data (non-targeted metabolomics of navy bean or rice bran powders) was used for confirmation of biological sample detected metabolite. Based on the presence of the metabolite in children and/or adults, and the magnitude of the fold-change/difference after consumption, metabolites were categorized into 3 tiers. Tier 1: Compound increased in both children and adults; with difference/change ≥2.0; p<0.05 Tier 2: Compound increased in either children or adults, with difference/change ≥2.0; p<0.05 Tier 3: Compound increased in either children or adults, with fold difference/change between 1.0 and 2.0; p<0.05 Using the above tiering criteria, 7, 17, and 36 compounds were categorized into Tiers 1, 2, and 3, respectively, for rice bran. Additionally, 5, 9, and 43 compounds were categorized into Tiers 1, 2, and 3, respectively, for navy bean. The final list of candidate compounds (Tier 1) were streamlined, based on (i) additional confirmation of metabolites using adult data from 2-weeks post-consumption, (ii) whether the metabolite has been detected in navy beans or rice bran in the literature (indicating either an existing non-validated dietary biomarker, or novel dietary biomarker), (ii) the specificity of compounds to navy beans or rice bran, (iii) cost and practicality considerations (e.g., availability and costs of authentic standards). A total of 11 candidate dietary biomarkers were selected for the targeted assayfor navy bean and rice bran that will be evaluated in the next reporting period.
Impacts
What was accomplished under these goals?
With regards to Aim 1, the targeted nutrient analysis on a series of meals/snacks that were prepared to include/excludecooked navy beans or rice bran were compared to the samecontrol meal/snack by an independent registered laboratory and these findings have also nowbeen confirmed with the database analysis outputs usingNutritionist Pro software analysis. Additionally, all the food metabolome analysis of the beans and rice bran separately, and following their inclusion into finished meals and snacks has been completed. These analyses andresults are currently being interpreted and visualized in graphs/tables and in preparation for manuscript submission/publication in 2018. For Aim 2, all of the blood, urine and stool metabolomics has been completed for 29 study participants from our BENEFIT study. Twomanuscripts werepublished regarding dietary, gut microbiome and stool metabolite outcomes from this trial in 2017, and anotherpublication in the British Journal of Nutrition(2017) highlights rice bran components identified instool that are considered candidate dietary markers of intake. There are two more papers in final stages of preparation for evaluating both rice bran and navy beans in the plasma and urine metabolomes of these study participants, as well as one focused on the navy bean food components in stool. The protocols for targeted food biomarker metabolite analysis in these biological matrices is underway and was a direct result of the non-targeted global metabolomics approach applied initially. A similar approach to Aim 2 was applied for Aim 3. One paper has been published for this aim that describes the nutritional outcomes and serum lipid results following dietary intervention: Borresen et. al. A Pilot Randomized Controlled Clinical Trial to Assess Tolerance and Efficacy of Navy Bean and Rice Bran Supplementation for Lowering Cholesterol in Children. Global Pediatric Health Volume 4: 1-10, 2017. Usingplasma samples collected from these children that participated in this clinical trial, we completedaplasma metabolome analysis that has been accepted for publication and is in press.Li et. al. Navy Bean and Rice Bran Intake Alters the Plasma Metabolome of Children at Risk for Cardiovascular Disease. Frontiers in Nutrition.doi: 10.3389/fnut.2017.00071, 2018. The targeted dietary metabolite biomarker analysis is underway for the remaining plasma samples available from these children and the results from this new protocol are anticipated to be available by end of 2018. Across the three specific aims: The integration of the global food metabolome for rice bran and navy beans with the plasma, urine and stool metabolome of people who ate fixed amounts of these foods is a novel approach, and our results thus far indicate promise to identify a set of dietary metabolites for these foods that can be used in future population based assessments that seek to correlate biomarkers of food intake with nutritional status and health outcomes.
Publications
- Type:
Journal Articles
Status:
Accepted
Year Published:
2017
Citation:
Brown DG, Borresen EC, Brown RJ, Ryan EP. Heat-stabilised rice bran consumption by colorectal cancer survivors modulates stool metabolite profiles and metabolic networks: a randomised controlled trial. British Journal of Nutrition. 2017 May;117(9):1244-1256. doi: 10.1017/S0007114517001106.
- Type:
Journal Articles
Status:
Accepted
Year Published:
2017
Citation:
Erica C. Borresen, MPH, NaNet Jenkins-Puccetti, MPH, Katie Schmitz, BSc, Dustin G. Brown, BSc, Austin Pollack, BSc, Amanda Fairbanks, BSc, Melissa Wdowik, PhD, RDN, Sangeeta Rao, PhD, Tracy L. Nelson, MPH, PhD, Gary Luckasen, MD and Elizabeth P. Ryan, PhD
A Pilot Randomized Controlled Clinical Trial to Assess Tolerance and Efficacy of Navy Bean and Rice Bran Supplementation for Lowering Cholesterol in Children. Global Pediatric Health
Volume 4: 110.2017. PubMed PMID: 28345013
- Type:
Journal Articles
Status:
Accepted
Year Published:
2017
Citation:
Zarei I, Brown DG, Nealon NJ, Ryan EP. Rice Bran Metabolome Contains Amino Acids, Vitamins & Cofactors, and Phytochemicals with Medicinal and Nutritional Properties. Rice (N Y). 2017;10: 24. PubMed PMID: 28547736.
- Type:
Journal Articles
Status:
Accepted
Year Published:
2016
Citation:
Erica C. Borresen, Dustin G. Brown, Greg Harbison, Lynn Taylor, Amanda Fairbanks, Joanne O'Malia, Marlon Bazan, Sangeeta Rao, Susan M. Bailey, Melissa Wdowik, Tiffany L. Weir, Regina J. Brown & Elizabeth P. Ryan. A Randomized Controlled Trial to Increase Navy Bean or Rice Bran Consumption in Colorectal Cancer Survivors. (2016): A Randomized Controlled Trial to Increase Navy Bean or Rice Bran Consumption in Colorectal Cancer Survivors, Nutrition and Cancer, DOI: 10.1080/01635581.2016.1224370
- Type:
Book Chapters
Status:
Accepted
Year Published:
2017
Citation:
Fermented Foods in Health and Disease Prevention: An overview. J.R. Wilburn EPR. In: Juana Frias CM-V, Elena Penas, editor. Fermented Foods in Health and Disease Prevention: Nikki Levy, 2017:3-19.
|
Progress 11/15/15 to 11/14/16
Outputs
Target Audience:In the first year, we have engaged twograduate students and one postdoc working across disciplines fromveterinary, toxicology, food and nutrition fields, andone undergraduate from in food scienc/food safety. We havetwo staff personell in the preparation of samples for metabolite profile analysis and they have completed preliminary analysis on experiments and objectives outlined in Aims 1, 2 and 3. Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided?This project has provided extensive training in food and nutritional metabolomics appraoches to both graduate students and undergraduates, and staff personnel. The integration of these techniques for evaluating dietary biomarkers of intake in children and adults is a novel undertaking and limited statistical methods have been applied in this context from randomized controlled clinical trials. we are working with Metabolon to peform semi qualitative metabolite analysis that can be follwed by targeted analysis for biomarker validation to intake levels. How have the results been disseminated to communities of interest?Findings from this research has been included into invited lectures given by Dr. Ryan to diverse scientific communiites, including but not limited to academic lectures at Michigan State Unviersity, a plenary talke at the American Association for Veterinary Nutritionists and Veterinary Diagnosticians meeting. This preliminary work from 2016 has been presented in the whole grains and healthy symposium at the annual American Chemical Society meeting. Additional research from this project has been presented by my students and staff at local meetings. What do you plan to do during the next reporting period to accomplish the goals?We will be continuing to analyze the plasma, urine and stool datasets from adults consuming the three dietary interventions. This data for serum will be compared to metabolite profiles from children consuming similar diets rich in rice bran and navy bean compared to control.
Impacts
What was accomplished under these goals?
In Aim I, we are examining the rice bran and navy bean metabolite profiles within a food matrix of meals and snacks that were developed for two human dietary intervention studies in children and adults. In this first progress report, the major activities that have been completed to achieve Aim I includes running GC/LC-MS metabolomics on study meals and snacks that have either rice bran or navy bean powder included or neither (control). This data will be used to characterize the metabolome of rice bran and navy bean food ingredients incorporated into these foods and consumed by study participants. Additionally, quantifiable proximate anlysis and fatty acid levels have been measured on a set of these study foods. Below is the methods and analysis strategy approach: Metabolomics: • Study foods were lyophilized and 100 grams were provided in a blinded manner to Metabolon for analysis. • Global metabolite profiling was performed by Metabolon using state-of-the-art UPLC-TOF MS and GC-MS platforms to enable broad metabolite coverage. • Study food samples were performed on a 96-well plate format using 80% methanol. • Data from platforms were processed using XCMS for peak detection, retention time alignment and normalization. • A list of all measured metabolites across study foods and groups were provided by Metabolon and values were normalized as raw area counts (original scale) as well as rescaled to set the median to 1 to assess metabolites that are higher and lower than the median. • The next step in the metabolomics analysis will be to review all measured metabolites across study groups for each study food to initiate a list of metabolites higher in rice bran and navy bean foods compared to control. Nutrient Composition: • Study foods were lyophilized and analyzed onsite at Warren Analytical Laboratory • The nutritional tests for proximate analysis included: o moisture by air oven drying AOAC 950.46 method, o protein by AOAC 992.15/992.23/992.15 (LECO) method o total fat by AOAC 945.44 method o ash by AOAC 920.153 method o total carbohydrates was estimated by difference calculation method (CH029.01), o dietary fibre by AOAC 991.43 method o calories were calculated using Atwater factors • The nutritional tests for fatty acids included: o saturated fat by AOAC 996.06 method o trans-fat by AOAC 996.06 method o Lauric by AOAC 996.06 method o Mytistol by AOAC 996.06 method o Palmitoleic by AOAC 996.06 method o Cis 11 Vaccenic Acid by AOAC 996.06 method o Trans Elaidic by AOAC 996.06 method o Trans Linoelaidic by AOAC 996.06 method o Gamma O6 GLA by AOAC 996.06 method o Eicosen by AOAC 996.06 method o Arachidonic - Omega 6 by AOAC 996.06 method o Behenic by AOAC 996.06 method o Docosahexaenoic - Omega by AOAC 996.06 method o Caproic by AOAC 996.06 method o Capric Acid by AOAC 996.06 method o Tridecanoic saturated by AOAC 996.06 method o Myristic by AOAC 996.06 method o Palmitic by AOAC 996.06 method o Stearic by AOAC 996.06 method o Cis 9 Oleic Acid by AOAC 996.06 method o Linoleic by AOAC 996.06 method o Cis O3 ALA by AOAC 996.06 method o Arachidic by AOAC 996.06 method o Eocosatrienoic Omega 3 by AOAC 996.06 method o EPA by AOAC 996.06 method o Erucic by AOAC 996.06 method o Lignoceric by AOAC 996.06 method o Caprylic Acid by AOAC 996.06 method • The next step in the nutrition composition analysis will be compare the proximate analysis and fatty acid levels across groups of each study food analyzed. For Aim 2, we will be assessing biomarkers of bean and/or rice bran intake in children that participated in a RCT. Methods & Analysis Strategy: - General study methods: o Participants: Children aged 8-13 years old (n=50) with elevated cholesterol were enrolled in a pilot, randomized controlled dietary intervention study (NCT01911390). o Interventions: Participants consumed muffins or a smoothie daily with no navy beans or rice bran (control, 0g), navy beans (17.5g), rice bran (15g), or a combination of navy bean (9g) and rice bran (8g) for 4 weeks. o Sample collection and analyses: Fasting blood samples were collected at baseline and week 4 and serum was sent to Metabolon (Durham, NC) for global metabolic profiling via ultra-performance liquid-chromatography tandem mass spectroscopy (UPLC-MS/MS). • 76 samples total (38 at baseline, 38 at week 4) were analyzed by Metabolon • Baseline: control (n=9), beans (n=10), rice bran (n=9), beans + bran (n=10) • Week 4: control (n=9), beans (n=10), rice bran (n=9), beans + bran (n=10) o Metabolites from the diets were analyzed by Metabolon against a total of 854 compounds of known identity o Metabolite ratios were calculated to identify metabolites that were significantly different between groups (fold-differences) or within groups (fold-changes) o Statistics: Repeated-measures ANOVA was used to analyze any statistically significant differences within- and between-groups (p≤0.05). - Analysis strategy: o From the dataset provided by Metabolon, statistically significant differences (increases or decreases) between the intervention groups at 4 weeks vs. control were isolated and analyzed o A literature search was performed on the significant metabolites to determine whether the metabolite could be a potential biomarker of intake, and/or to elucidate biological relevance/links to health o For significantly increased metabolites, their presence in the original food powders were confirmed (although not always possible) Results & General Conclusions: - Table 1 presents the potential biomarkers of intake for either beans or bran o Potential biomarkers of intake for beans include 6-oxopiperidine-2-carboxylate, trigonelline, 4-vinylphenol sulfate, and N-methylpipecholate o Potential biomarkers of intake for rice bran include pyridoxal o The metabolites gentisate and theanine were increased following consumption of the combination beans + bran, but not beans or bran alone - Table 2 presents statistically significantly increased or decreased metabolites following the consumption of beans, bran, or beans + bran intake for 4 weeks vs. control o Amino acids: Statistically significant decreases were observed in several amino acid metabolites following the consumption of beans, bran, and beans + bran. However, it should be noted that out of 183 total amino acid metabolites analyzed by Metabolon, only 7, 7, and 3 significant changes were observed in the beans, bran, and beans + bran groups, respectively, suggesting that the consumption of beans and/or bran did not have a significant effect on amino acid metabolites overall. o Lipids: Overall significant modulatory effects on lipid metabolites were observed following the consumption of beans and/or bran; however, patterns of lipid modulation were different between groups. Generally, consumption of beans alone increased lipid metabolites from fatty acid (dicarboxylate, acyl glycine, and acyl carnitine metabolism), lysolipid, and plasmalogen metabolism, whereas the consumption of bran alone increased lipid metabolites from fatty acid (monohydroxy) and sphingolipid metabolism, secondary bile acid metabolism, and ceramides that do not overlap with the lipid changes in the beans group. Consumption of the combination beans + bran resulted in increased metabolites from phospholipid metabolism, and some lysolipid metabolites, but overall the beans + bran combination group did not produce as many modulatory effects as beans or bran consumed alone (probably due to dose). o Other: Several significant decreases were observed in several nucleotides, cofactors & vitamins, and xenobiotics (do not appear to be associated with intake, no general pattern to observed significance)
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