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Hyperpolarized magnetic resonance spectroscopy (MRS) enables noninvasive, real\u2010time metabolic monitoring, while being able to distinguish between pathogenic bacterial metabolism and host metabolism. Using dDNP\u2010NMR, we demonstrate real\u2010time conversion of hyperpolarized [2\u2010<jats:sup>13<\/jats:sup>C]pyruvate to [1\u2010<jats:sup>13<\/jats:sup>C]acetate within <jats:italic>Shigella<\/jats:italic>\u2010infected HeLa cells. The conversion of hyperpolarized [1,4\u2010<jats:sup>13<\/jats:sup>C<jats:sub>2<\/jats:sub>]fumarate to [1,4\u2010<jats:sup>13<\/jats:sup>C<jats:sub>2<\/jats:sub>]malate was used to verify host cell integrity during infection, indicating that <jats:italic>Shigella<\/jats:italic> metabolism occurs within intact host cells. These findings highlight the potential of dDNP\u2010NMR to provide deeper insights into the metabolic dynamics of bacterial infections, offering a powerful tool for studying bacterial\u2010host interactions at a cellular level and facilitating the development of more precise therapeutic strategies for targeting infectious diseases.<\/jats:p>","DOI":"10.1002\/nbm.70154","type":"journal-article","created":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T07:14:22Z","timestamp":1760080462000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Real\u2010Time Intracellular Bacterial Metabolism During <i>Shigella<\/i> Infection Using Hyperpolarized [2\u2010<sup>13<\/sup>C]Pyruvate"],"prefix":"10.1002","volume":"38","author":[{"given":"Thomas\u00a0B.","family":"Wareham\u00a0Mathiassen","sequence":"first","affiliation":[{"name":"Center for Hyperpolarization in Magnetic Resonance, Department of Health Technology Technical University of Denmark  Kgs. 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