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Transient overexpression of hs GNMT and associated mutants was achieved with calcium phosphate transfection of the corresponding pcDNA3.3 mammalian overexpression plasmid, which was modified to include an internal ribosome entry site (IRES) downstream of the open reading frame to allow for bicistronic expression of GFP as a fluorescent marker of overexpression

Therefore, future research should incorporate detailed clinical data and adopt more longitudinal cohort designs to assess changes in fungal communities over time and their temporal relationship with disease progression

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