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The developed method is satisfied with the regulatory approach standards outlined in the ICH guidelines

The animals were anesthetized with Isofluran Baxter (IsoFlo, Abbot Laboratories) and intraperitoneally injected with ZRF mixture (Zolazepam 3.8 mg/ml, Tiletamin 3.8 mg/ml Xylazine 0.45 mg/ml and Fentanyl 2.6 g/ml) before transcardial perfusion with 4?C 2% dextran in 0.1 M phosphate buffer (PB) for approximately 30 seconds, immediately followed by 4% formaldehyde (FA) in PB for 10 minutes at the speed of 6 ml/min

Furthermore, TMAO induces EndMT and apoptosis through protein kinase RNA-like endoplasmic reticulum kinase (PERK) signaling, worsening plaque formation and cardiac function [121]

Neurology (2013) 9(3):14151

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