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After deparaffinization, sections were heated in an autoclave at 121 C for 5 min in 10 mM sodium citrate buffer (pH 6.0) or reacted with protease solution (Nichirei Biosciences, Tokyo, Japan) for epitope retrieval, after which endogenous peroxidase was inactivated by 0.3% hydrogen peroxide in methanol

Individuals with risk factors for B12 deficiency including strict vegans, those with gastrointestinal conditions, patients taking long-term metformin or proton pump inhibitors, and older adultsshould discuss screening with their GP, even in the absence of obvious symptoms

Vitamin B 12 deficiency is highly prevalent in older adults, particularly among those with impaired vitamin absorption due to conditions such as gastric surgery and atrophic gastritis ( 12 levels were predictive of short, medium, and long-term mortality at six, 12, and 48 months ( 12 levels were linked to higher all-cause and cardiovascular mortality during hospitalization