Au680 clinical chemistry analyzer

ProGN and proUGN concentrations in plasma and urine were quantified by using the Human Proguanylin ELISA and the Prouroguanylin Human ELISA kits (Cat. Nos. RD191046100R and RD191069200R, respectively; BioVendor a.s., Brno, Czech Republic). The ELISAs were performed according to the manufacturer’s instructions. The ELISA plate optical readings were done by using a SPECTRAmax microplate reader (Molecular Devices, Sunnyvale, CA, USA), and concentrations were calculated by using SoftMaxPro Software version 7.1 (Molecular Devices, Sunnyvale, CA, USA) using a curve fit based on its 4-parameter logistic nonlinear regression model.
Urine chloride, sodium, potassium, and protein levels were measured using the Cobas^® c 702 module (Roche, Basel, Switzerland); urine creatinine was measured using a AU680 Clinical Chemistry Analyzer (Beckman Coulter, Brea, CA, USA); and urine zinc was measured using inductively coupled plasma mass spectrometry (ICP-MS) on an ELAN DRC-e (PerkinElmer Inc., Waltham, MA, USA), all performed by the Laboratory Clinic at HUS.

Fasting blood samples (10 mL) were taken from IS patients and NCs within 72 h of recruitment, separated by centrifugation, and frozen at −80°C immediately after processing. Samples were shipped in packaging incorporating dry ice-cooling agents by courier from every site to a blood storage site, where they were stored at −80°C. Total cholesterol (TC), triglyceride (TG), low-density lipoprotein cholesterol (LDL-C), high-density lipoprotein cholesterol (HDL-C), apolipoprotein B (ApoB), ApoA1, fasting plasma glucose (FPG), high-sensitive C-reactive protein (hs-CRP), and homocysteine (Hcy) concentrations were measured in the laboratory of the Dian Diagnostics Group Co. Ltd with Beckman Coulter AU680 Clinical Chemistry Analyzer and Beckman reagents (Beckman Coulter, Brea, CA, USA). All the participants have not taken antibiotics, probiotics, and prebiotics in the 3 months prior to the feces collection. Feces were collected at hospital or home. The feces were collected according to the instruction and delivered immediately at low temperatures. The frozen feces were shipped using dry ice to Xiangxi Center for Disease Prevention and Control. Once received, fecal samples were divided into three parts of 200 mg and stored at −80°C until extraction.

A blood smear was prepared using whole blood for the evaluation of red blood cell morphology and platelet count. The remaining blood was aliquoted to a heparinized microhematocrit tube and a 1-milliliter lithium heparinized gel separator tube. The microhematocrit tube was centrifuged for 3 min in a BD Clay Adams™ TRIAC^® centrifuge, and the packed cell volume was immediately determined. Approximately 500 µL of plasma was collected following centrifugation at 1000× g for 15 min. Plasma was stored at −80 °C until the biochemical analysis was completed. The Louisiana State University’s Veterinary Clinical Pathology Service measured albumin, AST, blood urea nitrogen (BUN), and total bilirubin on a Beckman AU680 Clinical Chemistry Analyzer according to the manufacturer’s protocols. Lactate dehydrogenase (LDH) was measured using a colorimetric assay kit from Abcam™ (ab102526).

A mature model of partial hepatic warm IRI has been used [12 (link), 33 (link)]. Briefly, after successful anesthesia with isoflurane, heparin was injected into the mice. A midline laparotomy was performed, and an atraumatic clip was used to interrupt the arterial and portal venous blood supply to the cephalic lobes (70%) of the liver. After 90 min of partial hepatic warm ischemia, the clip was removed to initiate the process of hepatic reperfusion. Mice were maintained anesthetized with isoflurane and placed in the environment temperature at 26 °C. The mice were killed after 6 h of reperfusion and the collected samples were harvested for analysis. Sham controls underwent the same procedure but without vascular occlusion.
To study the effects of ROS, mice were pretreated with 300 mg/kg N-acetylcysteine (NAC) (Yeasen Biotechnology, Shanghai, China) or PBS by intraperitoneal injection.
Serum ALT and AST levels were measured using an AU680 clinical chemistry analyzer (Beckman Coulter, Brea, California, USA).
Liver specimens were fixed in 4% paraformaldehyde and embedded in paraffin for hematoxylin and eosin (H&E) and immunofluorescent staining. Some of the specimens were frozen in liquid nitrogen for 8-OHdG staining.

All biochemical procedures were done on the first day of hospital admission in a specialized biochemical laboratory of the Clinical Center Kragujevac, Serbia. Complete blood cell count (CBC) was measured using a hematology analyzer (DxH 800 Hematology Analyzer by Beckman Coulter). The biochemical parameters such as glucose, creatinine, urea, cholesterol, triglyceride (TG), aspartate aminotransferase (AST), alanine aminotransferase (ALT) gamma-glutamyl transferase (GGT), lactate dehydrogenase (LDH), total and direct bilirubin, C-reactive protein (CRP), sodium, and potassium were estimated from the serum samples by using standard kits in an automatic clinical chemistry analyzer (AU680 Clinical Chemistry Analyzer by Beckman Coulter). Measurement of the vitamin D level was performed using an automated immunoassay analyzer—the Alinity i system (Abbott Laboratories, IL, USA) that utilizes the chemiluminescent microparticle immunoassay (CMIA) principle. The level of procalcitonin in the serum was determined by the method of electrochemiluminescence, on the immunochemistry analyzer (Cobas e 411 by Roche). D dimer concentration measurement was performed on coagulation analyzer ACL-TOP 300 (Instrumentation Laboratory, Bedford, USA) employing the automated latex-enhanced particle immunoturbidimetric method.