{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"medRxiv"}],"indexed":{"date-parts":[[2026,5,9]],"date-time":"2026-05-09T07:14:21Z","timestamp":1778310861953,"version":"3.51.4"},"posted":{"date-parts":[[2025,7,16]]},"group-title":"Nutrition","reference-count":53,"publisher":"openRxiv","license":[{"start":{"date-parts":[[2025,7,16]],"date-time":"2025-07-16T00:00:00Z","timestamp":1752624000000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by-nc\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2025,7,16]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                <jats:sec>\n                  <jats:title>Background<\/jats:title>\n                  <jats:p>Choline is an essential nutrient, and insufficient intake negatively affects organs such as the liver, brain, and muscles. In the United States, average choline intake remains below the Adequate Intake (AI) (550 mg\/day men, 425 mg\/day women). Although conventional dietary assessment tools can identify people who are eating diets low in choline, no metabolite biomarkers have been proven to reliably assess choline intake.<\/jats:p>\n                <\/jats:sec>\n                <jats:sec>\n                  <jats:title>Objective<\/jats:title>\n                  <jats:p>We tested whether plasma concentrations of choline and its metabolites could determine dietary choline intake. We also assessed whether liver elastography (Fibroscan) could detect diet-induced changes in liver fat.<\/jats:p>\n                <\/jats:sec>\n                <jats:sec>\n                  <jats:title>Methods<\/jats:title>\n                  <jats:p>\n                    In a double-blind, randomized, crossover feeding study, participants adhered to 3 distinct dietary patterns for 2-wk intervals, delivering approximately 100%, 50%, and 25% of the choline AI. On Day 12 of each dietary arm, in addition to the food supplied, subjects consumed a single bolus of 2.2 mmol trimethyl-d\n                    <jats:sub>9<\/jats:sub>\n                    -choline. Plasma concentrations of choline, betaine and phosphatidylcholine (PtdCho) were measured using mass spectrometry. Targeted assays quantified choline, betaine, phosphatidylcholine and total homocysteine concentrations. Liver fat content was evaluated non-invasively using Fibroscan.\n                  <\/jats:p>\n                <\/jats:sec>\n                <jats:sec>\n                  <jats:title>Results<\/jats:title>\n                  <jats:p>\n                    Plasma concentrations of d\n                    <jats:sub>9<\/jats:sub>\n                    -choline, betaine, and their isotopic enrichment ratio (IER) varied with dietary intake (q&lt;0.0001), and PtdCho IER also differed significantly (q=0.001). In targeted analysis, choline and betaine concentrations were highly responsive to dietary choline intake, while PtdCho and tHcy were not. Receiver Operator Characteristic (ROC) analysis showed strong accuracy using plasma choline (AUC=0.81) and betaine (AUC=0.80), with improved accuracy when combined (AUC= 0.83). Fibroscan identified a subset of participants with increased liver fat in response to the 25% AI vs. 100% AI choline diet, though patterns varied among individuals.\n                  <\/jats:p>\n                <\/jats:sec>\n                <jats:sec>\n                  <jats:title>Conclusion<\/jats:title>\n                  <jats:p>Plasma choline and betaine concentrations are robust biomarkers of dietary choline intake under controlled feeding. These findings support targeted metabolite profiling to improve choline intake assessment and reveal induvial variability in liver response to low choline intake.<\/jats:p>\n                <\/jats:sec>","DOI":"10.1101\/2025.07.15.25331567","type":"posted-content","created":{"date-parts":[[2025,7,17]],"date-time":"2025-07-17T00:30:21Z","timestamp":1752712221000},"source":"Crossref","is-referenced-by-count":3,"title":["Choline and betaine concentrations in plasma predict dietary choline intake in healthy humans: a double-blind randomized control feeding study"],"prefix":"10.64898","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-4239-4795","authenticated-orcid":false,"given":"Isis","family":"Trujillo-Gonzalez","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"David A.","family":"Horita","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Julie","family":"Stegall","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Rachel","family":"Coble","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8344-818X","authenticated-orcid":false,"given":"Evan M.","family":"Paules","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Anju A.","family":"Lulla","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Emmanuel","family":"Baah","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4199-7551","authenticated-orcid":false,"given":"Teodoro","family":"Bottiglieri","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Wei","family":"Sha","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Martin","family":"Kohlmeier","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Walter B.","family":"Friday","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9282-161X","authenticated-orcid":false,"given":"Steven H.","family":"Zeisel","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"54368","reference":[{"key":"2025071804350427000_2025.07.15.25331567v1.1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1753-4887.2009.00246.x"},{"key":"2025071804350427000_2025.07.15.25331567v1.2","doi-asserted-by":"publisher","DOI":"10.1097\/NT.0000000000000302"},{"key":"2025071804350427000_2025.07.15.25331567v1.3","doi-asserted-by":"publisher","DOI":"10.1177\/0148607101025005260"},{"issue":"1","key":"2025071804350427000_2025.07.15.25331567v1.4","doi-asserted-by":"crossref","first-page":"163","DOI":"10.1093\/ajcn\/80.1.163","article-title":"Elevated serum creatine phosphokinase in choline-deficient humans: mechanistic studies in C2C12 mouse myoblasts","volume":"80","year":"2004","journal-title":"The American journal of clinical nutrition"},{"key":"2025071804350427000_2025.07.15.25331567v1.5","doi-asserted-by":"publisher","DOI":"10.1093\/advances\/nmac082"},{"key":"2025071804350427000_2025.07.15.25331567v1.6","unstructured":"Institute of Medicine Standing Committee on the Scientific Evaluation of Dietary Reference I, its Panel on Folate OBV, Choline. The National Academies Collection: Reports funded by National Institutes of Health. Dietary Reference Intakes for Thiamin, Riboflavin, Niacin, Vitamin B(6), Folate, Vitamin B(12), Pantothenic Acid, Biotin, and Choline. Washington (DC): National Academies Press (US) Copyright \u00a9 1998, National Academy of Sciences.; 1998."},{"issue":"1","key":"2025071804350427000_2025.07.15.25331567v1.7","doi-asserted-by":"crossref","first-page":"58","DOI":"10.1093\/advances\/nmx004","article-title":"Choline","volume":"9","year":"2018","journal-title":"Advances in nutrition"},{"issue":"6","key":"2025071804350427000_2025.07.15.25331567v1.8","doi-asserted-by":"crossref","first-page":"733","DOI":"10.1080\/19390211.2019.1639875","article-title":"Choline: The Neurocognitive Essential Nutrient of Interest to Obstetricians and Gynecologists","volume":"17","year":"2020","journal-title":"Journal of Dietary Supplements"},{"key":"2025071804350427000_2025.07.15.25331567v1.9","doi-asserted-by":"publisher","DOI":"10.3945\/ajcn.112.048462"},{"key":"2025071804350427000_2025.07.15.25331567v1.10","doi-asserted-by":"crossref","first-page":"445","DOI":"10.1016\/j.envres.2016.08.015","article-title":"Dietary micronutrient intake and its relationship with arsenic metabolism in Mexican women","volume":"151","year":"2016","journal-title":"Environ Res"},{"issue":"4","key":"2025071804350427000_2025.07.15.25331567v1.11","first-page":"642","article-title":"Dietary choline and folate relationships with serum hepatic inflammatory injury markers in Taiwanese adults","volume":"26","year":"2017","journal-title":"Asia Pac J Clin Nutr"},{"key":"2025071804350427000_2025.07.15.25331567v1.12","doi-asserted-by":"publisher","DOI":"10.1016\/j.jada.2006.07.020"},{"key":"2025071804350427000_2025.07.15.25331567v1.13","doi-asserted-by":"publisher","DOI":"10.1016\/j.jada.2008.10.046"},{"key":"2025071804350427000_2025.07.15.25331567v1.14","doi-asserted-by":"publisher","DOI":"10.1096\/fj.07-8227com"},{"key":"2025071804350427000_2025.07.15.25331567v1.15","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M110.106922"},{"key":"2025071804350427000_2025.07.15.25331567v1.16","doi-asserted-by":"publisher","DOI":"10.1096\/fj.09-154054"},{"key":"2025071804350427000_2025.07.15.25331567v1.17","doi-asserted-by":"publisher","DOI":"10.1093\/ajcn\/85.5.1275"},{"issue":"1","key":"2025071804350427000_2025.07.15.25331567v1.18","doi-asserted-by":"crossref","first-page":"35","DOI":"10.1038\/s42255-024-01203-8","article-title":"Cellular and organismal function of choline metabolism","volume":"7","year":"2025","journal-title":"Nature Metabolism"},{"key":"2025071804350427000_2025.07.15.25331567v1.19","doi-asserted-by":"publisher","DOI":"10.1038\/s41586-024-07444-7"},{"key":"2025071804350427000_2025.07.15.25331567v1.20","unstructured":"Service UAR . 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