Vettest ghp

Hemodynamic parameters were continuously monitored including MAP, mean pulmonary artery pressure (MPAP), right atrial pressure (RAP), cardiac output (CO), cardiac index (CI), and S_vO₂.
Blood was sequentially drawn for the determination of (i) blood gases, (ii) arterial lactate, (iii) plasma concentration of urea, creatinine, albumin (VetTest GHP, Idexx, Saint-Denis, France) (iv) blood cell count, and (v) TNF-α, and IL-6 (ELISA, RnD Systems, Minneapolis, USA). At the end of the experiment, bacterial counts were performed on blood samples, and lung and kidney biopsies were taken for histological analyses. Histology scores were determined by an experienced pathologist who was blinded to the treatment applied.

Organ dysfunction was determined by measurement of biochemical indicators of organ functions in plasma samples and by histology scores. Plasma concentrations of urea and creatinine (indicators of renal dysfunction), alanine aminotransferase (ALT) and alkaline phosphatase (ALKP) (indicators of liver dysfunction), and amylase and glycemia (indicators of pancreatic dysfunction) were analyzed by an automate (VetTest GHP, Idexx, Saint-Denis, France) after the animals’ sacrifice 2 or 7 days after the CLP procedure.
Forty-eight hours after CLP, the lungs, liver, kidneys, and spleen were fixed in 4% paraformaldehyde during 24 h at 4 °C, dehydrated through a series of ethanol concentrations, cleared with toluene, embedded in paraffin wax, and cut into 5-mm-thick sections with a Leica micro-tome (RM2135, Leica, France). For histological examination, each specimen was stained with hematoxylin-erythrosin-saffron, mounted on glass slides, and visualized on an optical microscope (DMD 108, Leica, France).
Histology scores were performed by an experienced pathologist blinded to the treatment administered. The scoring system was adapted from [34 (link), 35 (link)] and ranged from 0 to 9 for the lungs, 0 to 5 for the kidneys, 0 to 4 for the liver, and 0 to 3 for the spleen.