Sugar, albumin, creatinine, urea, potassium, and sodium levels were estimated using the specified kits according to the instructions of the supplier.
All biochemical analyses (lipid profile tests "sTC, sTG, sLDL-C, sHDL-C, and sVLDL-C," liver enzymes "AST, ALT, and ALP," and kidney function parameters "creatinine, urea, and uric acid") were achieved using the specified kits from Gesellschaft for Biochemical and Diagnostic (Germany) according to the instructions of the suppliers.
Lipid peroxide was estimated by measuring malondialdehyde (MDA) and the activity of catalase, reduced glutathione (GSH), and superoxide dismutase (SOD) in the serum and in the kidney tissue homogenate using the specified kits from Biodiagnostic Chemical Company (Egypt) according to the instructions of the supplier.
DNA extraction was done with the concentrated pellet using MagNA Pure LC instruments (Roche, USA) with MagNA Pure LC DNA isolation kits III (Roche, USA) following automated mode specified by the kit manufacturer's instructions manual.
Amounts of the cysLTs C4, D4 and E4 (as determined with the enzyme immunoassay kit specified in the following paragraph) were below the detection limit.
The reaction condition used was as specified in the kit, with an annealing temperature of 55°C and 45 cycles for the data collection step.
Blank control was performed without the addition of protein as specified by the kit and the degree of enrichment is calculated by subtracting with blank control.
Lysis solution (DNeasy® kit buffer AP1 containing 25 mg mL−1 polyvinylpolypyrrolidone), along with RNase A, was added to the powdered leaf samples and the mixture was incubated at 65 °C, as specified in the kit instructions.
Before LC3 detection, to assess autophagic flux, cells were incubated for 2 hr with the autophagy flux inhibitor provided by the kit specified and according to the manufacturer's instructions.
To evaluate the accuracy of the molecular detection compared to conventional culture, we considered only concordance with those samples with positive cultures for any of the organisms specified in the kit.
