{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,6]],"date-time":"2026-07-06T15:15:33Z","timestamp":1783350933027,"version":"3.54.6"},"reference-count":44,"publisher":"MDPI AG","issue":"13","license":[{"start":{"date-parts":[[2026,7,4]],"date-time":"2026-07-04T00:00:00Z","timestamp":1783123200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"This research received no external funding."}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["JCM"],"abstract":"<jats:p>Background: Reliable intraoperative tools for donor liver assessment are needed, particularly in the context of steatotic and extended-criteria grafts. While histology remains the reference standard, it is limited by sampling variability and logistical constraints. Preservation fluid may provide a complementary, whole-organ source of biochemical information. Methods: In this single-center prospective exploratory pilot study, liver tissue and preservation fluid were collected from 30 donation-after-brain-death grafts during the back-table procedure. Biochemical parameters, including arginase activity, \u03b2-hydroxybutyrate (\u03b2HB), acetoacetate, and total protein, were measured using standard assays. Associations with histological steatosis on wedge biopsy were assessed using nonparametric correlation analyses, and paired preservation fluid samples were compared. Results: Most grafts demonstrated absent or mild steatosis; only two exhibited moderate steatosis, and none were severely steatotic. No preservation fluid biomarker showed a statistically significant association with histological steatosis. Weak, non-significant positive correlations were observed for \u03b2HB and arginase activity (Spearman r \u2248 0.33\u20130.35). Protein concentration and arginase activity decreased between start and end samples, whereas ketone body levels remained relatively stable. Conclusions: Preservation fluid biomarker measurement during routine graft preparation is feasible. Although no significant associations with histological steatosis were identified, the observed weak correlations suggest possible associations requiring validation in larger studies. Larger, adequately powered studies, including a broader spectrum of steatosis and clinically relevant outcomes, are required to determine potential clinical applicability.<\/jats:p>","DOI":"10.3390\/jcm15135239","type":"journal-article","created":{"date-parts":[[2026,7,6]],"date-time":"2026-07-06T13:43:14Z","timestamp":1783345394000},"page":"5239","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Exploratory Analysis of Liver Tissue and Preservation Fluid Biomarkers (\u03b2-Hydroxybutyrate and Arginase) in Relation to Graft Steatosis"],"prefix":"10.3390","volume":"15","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-1325-7094","authenticated-orcid":false,"given":"Kawthar","family":"Safi","sequence":"first","affiliation":[{"name":"Department of Biochemistry, Medical University of Warsaw, 02-097 Warsaw, Poland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0003-0751-2818","authenticated-orcid":false,"given":"Angelika Joanna","family":"Pawlicka","sequence":"additional","affiliation":[{"name":"Department of Biochemistry, Medical University of Warsaw, 02-097 Warsaw, Poland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Gra\u017cyna","family":"Kubiak-Tomaszewska","sequence":"additional","affiliation":[{"name":"Department of Biochemistry, Medical University of Warsaw, 02-097 Warsaw, Poland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0181-3607","authenticated-orcid":false,"given":"Marta","family":"Struga","sequence":"additional","affiliation":[{"name":"Department of Biochemistry, Medical University of Warsaw, 02-097 Warsaw, Poland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7851-7147","authenticated-orcid":false,"given":"Andriy","family":"Zhylko","sequence":"additional","affiliation":[{"name":"Department of General, Transplant and Liver Surgery, Medical University of Warsaw, Banacha 1A, 02-097 Warsaw, Poland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Maciej","family":"Krasnod\u0119bski","sequence":"additional","affiliation":[{"name":"Department of General, Transplant and Liver Surgery, Medical University of Warsaw, Banacha 1A, 02-097 Warsaw, Poland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Micha\u0142","family":"Gr\u0105t","sequence":"additional","affiliation":[{"name":"Department of General, Transplant and Liver Surgery, Medical University of Warsaw, Banacha 1A, 02-097 Warsaw, Poland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6928-4561","authenticated-orcid":false,"given":"Alicja","family":"Chrzanowska","sequence":"additional","affiliation":[{"name":"Department of Biochemistry, Medical University of Warsaw, 02-097 Warsaw, Poland"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2026,7,4]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"242","DOI":"10.12659\/AOT.914246","article-title":"Risk Factors for High Mortality on the Liver Transplant Waiting List in Times of Organ Shortage: A Single-Center Analysis","volume":"24","author":"Husen","year":"2019","journal-title":"Ann. Transplant."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"101217","DOI":"10.1016\/j.jceh.2023.07.004","article-title":"Donor Evaluation Protocol for Live and Deceased Donors","volume":"14","author":"Puri","year":"2024","journal-title":"J. Clin. Exp. Hepatol."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"1040","DOI":"10.1016\/j.jhep.2024.07.032","article-title":"EASL Clinical Practice Guidelines on Liver Transplantation","volume":"81","author":"Samuel","year":"2024","journal-title":"J. Hepatol."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"4","DOI":"10.1007\/s40137-024-00434-8","article-title":"\u201cHope and Hurdles\u201d in Liver Machine Perfusion: A Review and Future Directions of Machine Perfusion in Liver Transplantation","volume":"13","author":"Guarrera","year":"2024","journal-title":"Curr. Surg. Rep."},{"key":"ref_5","doi-asserted-by":"crossref","unstructured":"Abbas, S.H., Ceresa, C.D.L., and Pollok, J.M. (2024). Steatotic Donor Transplant Livers: Preservation Strategies to Mitigate against Ischaemia-Reperfusion Injury. Int. J. Mol. Sci., 25.","DOI":"10.3390\/ijms25094648"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"405","DOI":"10.1097\/TP.0000000000004291","article-title":"Impact of Donor Liver Macrovesicular Steatosis on Deceased Donor Yield and Posttransplant Outcome","volume":"107","author":"Kwong","year":"2023","journal-title":"Transplantation"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"100091","DOI":"10.1016\/j.liver.2022.100091","article-title":"Current Techniques and Future Trends in the Diagnosis of Hepatic Steatosis in Liver Donors: A Review","volume":"7","author":"Rajamani","year":"2022","journal-title":"J. Liver Transplant."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"1999","DOI":"10.1053\/j.gastro.2019.11.312","article-title":"MAFLD: A Consensus-Driven Proposed Nomenclature for Metabolic Associated Fatty Liver Disease","volume":"158","author":"Eslam","year":"2020","journal-title":"Gastroenterology"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"1542","DOI":"10.1016\/j.jhep.2023.06.003","article-title":"A Multisociety Delphi Consensus Statement on New Fatty Liver Disease Nomenclature","volume":"79","author":"Rinella","year":"2023","journal-title":"J. Hepatol."},{"key":"ref_10","doi-asserted-by":"crossref","unstructured":"Mooli, R.G.R., and Ramakrishnan, S.K. (2022). Emerging Role of Hepatic Ketogenesis in Fatty Liver Disease. Front. Physiol., 13.","DOI":"10.3389\/fphys.2022.946474"},{"key":"ref_11","doi-asserted-by":"crossref","unstructured":"Kwon, S., Jeyaratnam, R., and Kim, K.H. (2024). Targeting Ketone Body Metabolism to Treat Fatty Liver Disease. J. Pharm. Pharm. Sci., 27.","DOI":"10.3389\/jpps.2024.13375"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"1231","DOI":"10.1016\/j.jhep.2013.07.034","article-title":"MicroRNA Profiles in Graft Preservation Solution Are Predictive of Ischemic-Type Biliary Lesions after Liver Transplantation","volume":"59","author":"Verhoeven","year":"2013","journal-title":"J. Hepatol."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"1361","DOI":"10.1053\/j.gastro.2017.12.027","article-title":"Glycome Patterns of Perfusate in Livers Before Transplantation Associate with Primary Nonfunction","volume":"154","author":"Verhelst","year":"2018","journal-title":"Gastroenterology"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"479","DOI":"10.1097\/SLA.0000000000006822","article-title":"A New Era of Decision-Making in Liver Transplantation: A Prospective Validation and Cost-Effectiveness Analysis of FMN-Guided Liver Viability Assessment During Normothermic Machine Perfusion","volume":"282","author":"Wehrle","year":"2025","journal-title":"Ann. Surg."},{"key":"ref_15","doi-asserted-by":"crossref","unstructured":"Pali, C., Kim, D.V., Mantik, S., Lee, K.E.K., So, J.-B., Moon, C.-Y., Park, S., Kwak, D.-H., Kang, H.-B., and Pali, D.V. (2025). Unraveling the Translational Relevance of \u03b2-Hydroxybutyrate as an Intermediate Metabolite and Signaling Molecule. Int. J. Mol. Sci., 26.","DOI":"10.3390\/ijms26157362"},{"key":"ref_16","doi-asserted-by":"crossref","unstructured":"Clemente, G.S., van Waarde, A., Antunes, I.F., D\u00f6mling, A., and Elsinga, P.H. (2020). Arginase as a Potential Biomarker of Disease Progression: A Molecular Imaging Perspective. Int. J. Mol. Sci., 21.","DOI":"10.3390\/ijms21155291"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"2069","DOI":"10.1016\/j.cmet.2024.07.007","article-title":"Hierarchical Tricarboxylic Acid Cycle Regulation by Hepatocyte Arginase 2 Links the Urea Cycle to Oxidative Metabolism","volume":"36","author":"Zhang","year":"2024","journal-title":"Cell Metab."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"20405","DOI":"10.1038\/srep20405","article-title":"Targeting Arginase-II Protects Mice from High-Fat-Diet-Induced Hepatic Steatosis through Suppression of Macrophage Inflammation","volume":"6","author":"Liu","year":"2016","journal-title":"Sci. Rep."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"1207","DOI":"10.1016\/j.jhep.2014.12.036","article-title":"Arginase 1: A Potential Marker of a Common Pattern of Liver Steatosis in HCV and NAFLD Children","volume":"62","author":"Alisi","year":"2015","journal-title":"J. Hepatol."},{"key":"ref_20","doi-asserted-by":"crossref","unstructured":"Basta, G., Babboni, S., Pezzati, D., Del Turco, S., Balzano, E., Catalano, G., Russo, L., Tincani, G., Carrai, P., and Petruccelli, S. (2025). Perfusate Liver Arginase 1 Levels After End-Ischemic Machine Perfusion Are Associated with Early Allograft Dysfunction. Biomedicines, 13.","DOI":"10.3390\/biomedicines13010244"},{"key":"ref_21","unstructured":"(2025, December 28). Cell Membrane\u2014Servier Medical Art. Available online: https:\/\/smart.servier.com\/category\/cellular-biology\/cell-membrane\/."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"766","DOI":"10.1159\/000522052","article-title":"Primary Nonfunction and Early Allograft Dysfunction after Liver Transplantation","volume":"40","author":"Masior","year":"2022","journal-title":"Dig. Dis."},{"key":"ref_23","first-page":"219","article-title":"Assessing Donor Liver Quality and Restoring Graft Function in the Era of Extended Criteria Donors","volume":"11","author":"Lin","year":"2022","journal-title":"J. Clin. Transl. Hepatol."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"147","DOI":"10.1172\/JCI200422422","article-title":"Molecular Mediators of Hepatic Steatosis and Liver Injury","volume":"114","author":"Browning","year":"2004","journal-title":"J. Clin. Investig."},{"key":"ref_25","doi-asserted-by":"crossref","unstructured":"Ichim, C., Boicean, A., Todor, S.B., Anderco, P., and B\u00eerlu\u021biu, V. (2025). Fecal Microbiota Transplantation in Patients with Alcohol-Associated Cirrhosis: A Clinical Trial. J. Clin. Med., 14.","DOI":"10.3390\/jcm14175981"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"6706","DOI":"10.1074\/jbc.M109.064501","article-title":"A Sequence Variation (I148M) in PNPLA3 Associated with Nonalcoholic Fatty Liver Disease Disrupts Triglyceride Hydrolysis","volume":"285","author":"He","year":"2010","journal-title":"J. Biol. Chem."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"84","DOI":"10.4239\/wjd.v12.i1.84","article-title":"Novel Glucose-Lowering Drugs for Non-Alcoholic Fatty Liver Disease","volume":"12","author":"Fu","year":"2021","journal-title":"World J. Diabetes"},{"key":"ref_28","doi-asserted-by":"crossref","unstructured":"Fougerat, A., Montagner, A., Loiseau, N., Guillou, H., and Wahli, W. (2020). Peroxisome Proliferator-Activated Receptors and Their Novel Ligands as Candidates for the Treatment of Non-Alcoholic Fatty Liver Disease. Cells, 9.","DOI":"10.3390\/cells9071638"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"2565","DOI":"10.2147\/DDDT.S307257","article-title":"Metformin in Combination with Malvidin Prevents Progression of Non-Alcoholic Fatty Liver Disease via Improving Lipid and Glucose Metabolisms, and Inhibiting Inflammation in Type 2 Diabetes Rats","volume":"15","author":"Zou","year":"2021","journal-title":"Drug Des. Devel. Ther."},{"key":"ref_30","doi-asserted-by":"crossref","unstructured":"Meszaros, A.T., Weissenbacher, A., Schartner, M., Egelseer-Bruendl, T., Hermann, M., Unterweger, J., Mittelberger, C., Reyer, B.A., Hofmann, J., and Zelger, B.G. (2024). The Predictive Value of Graft Viability and Bioenergetics Testing Towards the Outcome in Liver Transplantation. Transpl. Int., 37.","DOI":"10.3389\/ti.2024.12380"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"1217","DOI":"10.1007\/s00216-015-9227-8","article-title":"Extending Metabolome Coverage for Untargeted Metabolite Profiling of Adherent Cultured Hepatic Cells","volume":"408","author":"Castell","year":"2016","journal-title":"Anal. Bioanal. Chem."},{"key":"ref_32","doi-asserted-by":"crossref","unstructured":"Xu, J., Hassan-Ally, M., Casas-Ferreira, A.M., Suvitaival, T., Ma, Y., Vilca-Melendez, H., Rela, M., Heaton, N., Wayel, J., and Legido-Quigley, C. (2020). Deregulation of the Purine Pathway in Pre-Transplant Liver Biopsies Is Associated with Graft Function and Survival after Transplantation. J. Clin. Med., 9.","DOI":"10.3390\/jcm9030711"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"17737","DOI":"10.1038\/srep17737","article-title":"Lipidomics Comparing DCD and DBD Liver Allografts Uncovers Lysophospholipids Elevated in Recipients Undergoing Early Allograft Dysfunction","volume":"5","author":"Xu","year":"2015","journal-title":"Sci. Rep."},{"key":"ref_34","doi-asserted-by":"crossref","unstructured":"Liu, Z., Zhu, H., Wang, W., Xu, J., Que, S., Zhuang, L., Qian, J., Wang, S., Yu, J., and Zhang, F. (2020). Metabonomic Profile of Macrosteatotic Allografts for Orthotopic Liver Transplantation in Patients with Initial Poor Function: Mechanistic Investigation and Prognostic Prediction. Front. Cell Dev. Biol., 8.","DOI":"10.3389\/fcell.2020.00826"},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"50","DOI":"10.2337\/diab.37.1.50","article-title":"Lipogenesis from Ketone Bodies in Perfused Livers from Streptozocin-Induced Diabetic Rats","volume":"37","author":"Freed","year":"1988","journal-title":"Diabetes"},{"key":"ref_36","doi-asserted-by":"crossref","unstructured":"Raigani, S., Karimian, N., Huang, V., Zhang, A.M., Beijert, I., Geerts, S., Nagpal, S., Hafiz, E.O.A., Fontan, F.M., and Aburawi, M.M. (2020). Metabolic and Lipidomic Profiling of Steatotic Human Livers during Ex Situ Normothermic Machine Perfusion Guides Resuscitation Strategies. PLoS ONE, 15.","DOI":"10.1371\/journal.pone.0228011"},{"key":"ref_37","doi-asserted-by":"crossref","unstructured":"Verstraeten, L., and Jochmans, I. (2022). Sense and Sensibilities of Organ Perfusion as a Kidney and Liver Viability Assessment Platform. Transpl. Int., 35.","DOI":"10.3389\/ti.2022.10312"},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1042\/bj3360001","article-title":"Arginine Metabolism: Nitric Oxide and Beyond","volume":"336","author":"Wu","year":"1998","journal-title":"Biochem. J."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"641","DOI":"10.1152\/physrev.00037.2016","article-title":"Arginase: A Multifaceted Enzyme Important in Health and Disease","volume":"98","author":"Caldwell","year":"2018","journal-title":"Physiol. Rev."},{"key":"ref_40","doi-asserted-by":"crossref","unstructured":"Moon, J., Do, H.J., Cho, Y., and Shin, M.J. (2014). Arginase Inhibition Ameliorates Hepatic Metabolic Abnormalities in Obese Mice. PLoS ONE, 9.","DOI":"10.1371\/journal.pone.0103048"},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"1602S","DOI":"10.1093\/jn\/137.6.1602S","article-title":"Arginine Metabolism: Boundaries of Our Knowledge","volume":"137","author":"Morris","year":"2007","journal-title":"J. Nutr."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"395","DOI":"10.1016\/j.tips.2015.03.006","article-title":"Arginase: An Old Enzyme with New Tricks","volume":"36","author":"Caldwell","year":"2015","journal-title":"Trends Pharmacol. Sci."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"467","DOI":"10.1089\/bio.2022.0053","article-title":"The Influence of Preanalytical Biospecimen Handling on the Measurement of B Vitamers, Amino Acids, and Other Metabolites in Blood","volume":"21","author":"Michels","year":"2023","journal-title":"Biopreserv. Biobank."},{"key":"ref_44","doi-asserted-by":"crossref","unstructured":"Thachil, A., Wang, L., Mandal, R., Wishart, D., and Blydt-Hansen, T. (2024). An Overview of Pre-Analytical Factors Impacting Metabolomics Analyses of Blood Samples. Metabolites, 14.","DOI":"10.3390\/metabo14090474"}],"container-title":["Journal of Clinical Medicine"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2077-0383\/15\/13\/5239\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,7,6]],"date-time":"2026-07-06T14:34:55Z","timestamp":1783348495000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2077-0383\/15\/13\/5239"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,7,4]]},"references-count":44,"journal-issue":{"issue":"13","published-online":{"date-parts":[[2026,7]]}},"alternative-id":["jcm15135239"],"URL":"https:\/\/doi.org\/10.3390\/jcm15135239","relation":{},"ISSN":["2077-0383"],"issn-type":[{"value":"2077-0383","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,7,4]]}}}