Morphometric Study
The morphometric study was conducted by two independent histopathologists experienced in animal models. They were not aware of which animal group the images come from and analyzed each criterion separately. Lesions identified in each organ were described according to the International Harmonization of Nomenclature and Diagnostic Criteria (INHAND) (16, 17).
A semiquantitative scoring analyses of electron microscope images was used to evaluate the degree of ultrastructural damage, as previously described by Sergazy et al., 2020, which utilizes a four-point scale (Grade 1–4) for the morphological assessment of subcellular structures (18, 19). In brief, slides assessed for different morphologic changes, and lesions were classified as present/absent on a scale from 0 to 1, or assessed for severity on a scale from 0 to 4. Histopathological lesions of the heart evaluated as present – ”1” or absent – ”0”, included cardiomyocyte hypertrophy, focal hyper-eosinophilia, enlargement of cardiomyocyte nuclei/multinucleated cardiomyocytes, cellular infiltrate, and atherosclerosis. Fibrosis (perivascular and interstitial) was assessed on a scale: ”0” – none, ”1” – minimal changes, ”2” – mild changes, ”3” – moderate changes, ”4” – severe lesions. Kidney lesions evaluated as present – ”1” or absent – ”0”, included tubular epitheliocyte vacuolization and hyperplasia, formation of protein cylinders in tubules, interstitial cellular infiltrate, thickening and fibrosis of glomerular membranes, hyaline glomerulopathy, and glomerulosclerosis. Liver lesions evaluated as present – ”1” or absent – ”0”, included focal hepatocyte vacuolization, presence of hyperchromatic/double-nucleated hepatocytes, lymphocytic infiltration, hyperplasia (cysts) of bile ducts, and periportal fibrosis. Glycogen accumulation was differentiated on a scale from 0 to 3: ”0” – less than 10% of hepatocytes, ”1” – 11-30% of hepatocytes, ”2” – 31-60%, ”3” – more than 61% of hepatocytes.