Cystatin c

In the first experiment (28 day follow-up), urea nitrogen (urea), glucose, lactate dehydrogenase (LDH) and alanine transaminase (ALT) were analyzed in plasma samples with Cobas c111 analyzer (Roche, Basel, Switzerland). The lower detection limit for urea was 3 mg/dl, for glucose 1.98 mg/dl, for LDH 10U/l, and for ALT was 2U/l. In the second experiment (BT-based outcome stratification), cystatin C (Biovendor, Karasek, Czech Republic) and troponin I (Life Diagnostics, West Chester, PA, USA) were analyzed in plasma (obtained from undiluted whole blood samples) using ELISA kits according to manufacturer's recommendations. Creatinine was analyzed with Cobas c111 analyzer using the same plasma samples. The lower detection limit for cystatin C was 0.04 ng/ml, for troponin 0.156 ng/ml. In the context of the liver, it must be noted that ALT and LDH are indicators of hepatocyte and general (LDH) tissue injury, thus, they do not per se define the level of liver dysfunction (although the latter effect typically develops once a pronounced injury occurs - as in CLP-ODam mice). Given that the parameters we recorded in this study are indicative of either organ injury or (dys)function, we refer to those data by the ‘organ injury/dysfunction’ acronym.

Albuminuria was assessed by ELISA according to the manufacturer’s instructions (Bethyl Laboratories, Inc., Montgomery, TX, United States) (Watson et al., 2012 (link)).
Cystatin C (BioVendor – Laboratorni medicina a.s., Karasek, Brno, Czech Republic) and KIM-1 (kidney injury molecule-1, or hepatitis A virus cellular receptor 1; Assay Matrix; EIAab, Optics Valley, Wuhan, China) ELISA analyses were carried out on plasma samples as per the manufacturer’s instructions.