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Although the actual mechanisms of MTX-induced liver and kidney damage are unknown, one of the hypotheses is the breakdown in cellular antioxidant defences, which increases the production of reactive oxygen species and nitrogen species, inhibits cytosolic NADP-dependent dehydrogenase and NADP malic enzyme, lowers glutathione (GSH), superoxide dismutase (SOD) and catalase levels and reduces the effectiveness of the antioxidant defence system protecting the cell against reactive oxygen species (Reference Savran, Cicek and Doguc7)

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The amino acid driven alteration of diversity and composition of the GM can also be modulated by the different types of metabolites produced, including nitric oxide, ammonia, polyamines, hydrogen sulfide (H 2 S), indoles, and phenols in both the proximal and distal intestine (Dai et al., 2011