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Item A High Serum Level of Taurocholic Acid is Correlated with the Severity and Resolution of Drug-induced Liver Injury(Elsevier, 2020) Tian, Qiuju; Yang, Ruiyuan; Wang, Yan; Liu, Jimin; Wee, Aileen; Saxena, Romil; Wang, Lan; Li, Min; Liu, Liwei; Shan, Shan; Kong, Yuanyuan; Ma, Hong; Ou, Xiaojuan; You, Hong; Zhao, Xinyan; Jia, Jidong; Pathology and Laboratory Medicine, School of MedicineBackground & Aims Alterations in the serum levels of bile acids are associated with drug-induced liver injury (DILI). We investigated the association between serum levels of bile acids and the severity and outcome of DILI, along with the potential role of variants in the ATP binding cassette subfamily B member 11 ( ABCB11) gene and expression of its product, ABCB11 (also called BSEP). Methods We performed this prospective study of 95 patients (median age, 53 years; 73.7% female) with DILI from August 2018 through August 2019. Patients were matched for age, gender, and body mass index with healthy individuals (n=100; healthy controls) and patients with chronic hepatitis B (n=105; CHB controls). We collected demographic and biochemical data at baseline and 1 week, 1 month, 3 months, and 6 months after DILI onset and at the time of biochemical recovery, liver failure or liver transplantation. Serum levels of bile acids were measured using high-performance liquid-chromatography tandem mass-spectrometry. All 27 exons of ABCB11 were sequenced and expression of BSEP were analyzed by immunohistochemistry in liver biopsy specimens. Results Levels of 30 of the 37 bile acids analyzed differed significantly between patients with DILI and healthy controls. Changes in levels of taurocholic acid (TCA), glycocholic acid, taurochenodeoxycholate, and glycochenodeoxycholate associated with the increased levels of bilirubin and greater severity of DILI, and were also associated with CHB. Cox regression analysis showed that only change in the levels of TCA independently associated with biochemical resolution of DILI. Combination of TCA level (≥ 1955.41 nmol/L), patient age, and DILI severity was associated with abnormal blood biochemistry at 6 months after DILI onset (area under the curve, 0.81; 95% confidence interval, 0.71–0.88; sensitivity, 0.69; specificity, 0.81). ABCB11 missense variants were not associated with differences in the serum bile acid profiles, DILI severity, or clinical resolution. However, lower levels of BSEP in bile canaliculi in liver biopsies were associated with altered serum levels of bile acids. Conclusions In this prospective study performed in Chinese patients, we found that the serum levels of TCA were associated with the severity and clinical resolution of DILI. Reduced protein expression of BSEP in liver tissue, rather than variants of the ABCB11 gene were associated with altered serum levels of bile acids.Item Identifying Metabolic Pathways Producing Alkamides in Echinacea purpurea(Office of the Vice Chancellor for Research, 2016-04-08) Williams, Jermell; Teitgen, Alicen; Minto, RobertEchinacea purpurea is a widely used herbal supplement that is frequently taken to relieve cold symptoms. Alkamides are a secondary metabolite found throughout the Echinacea genus that contain fatty acid chains incorporated into amides and are believed to be the bioactive agent in Echinacea. Our goal is to identify and understand the specific metabolic processes by which E. purpurea produces alkamides. In our experiment, Echinacea seedlings were grown to where the first true leaf emerged and unfurled which is when alkamide production is known to be most active. Alkamides were then extracted and taken to the GC/MS and LC/MS for analysis. Extracted alkamides were analyzed by triplequadrupole chromatography to investigate 13C labeling by glucose. Solid phase extractions were also performed to better observe fragmentation patterns. Fatty acids were also extracted to determine if fatty acids and alkamides were affected the same way by light or the lack of light, which would indicate that they are being synthesized in the same place. It was determined that neither compound experienced a synthesis decrease in the dark significant enough to support a model where acyl chains are newly created in the chloroplasts. Therefore alkamides are more likely to be made in the mitochondria. We are currently in the process of examining the spectra in order to determine the structures of the alkamides as well as any metabolic relationships.Item Identifying Metabolic Pathways Producing Alkamides in Echinacea purpurea(Office of the Vice Chancellor for Research, 2015-04-17) Williams, Jermell; Teitgen, Alicen; Minto, Robert E.Echinacea purpurea is a widely used herbal supplement that is frequently taken to relieve cold symptoms; alkamides are believed to be the bioactive agent. Alkamides are natural products found throughout the Echinacea genus that contain fatty acid chains incorporated into amides. Our goal is to identify and understand the specific metabolic processes by which E. purpurea produces alkamides. In our experiment Echinacea seedlings were grown to the point where the first true leaf started to grow and alkamide production is known to be active. Alkamides were then extracted and taken to the GC/MS for analysis. Extracted alkamides were analyzed by triple-quadrupole chromatography to investigate 13C labeling by glucose. We are currently in the process of examining the spectra in order to determine the structures of the alkamides as well as any metabolic relationships and if these are altered by a lack of light.