To regulate SAA content of the diets, capsules with (SAAhigh) or without (SAAlow) methionine and cysteine will be provided in a double-blind manner. The SAAlow group will receive capsules with no SAA (3000 mg/days maltodextrin) (capsules: Capsuline, Florida, USA; maltodextrin: (Star Nutrition, Sweden), whereas the SAAhigh group will receive capsules with methionine (1125 mg/days) and cystine (2500 mg/days) (Jo Mar Laboratories, Scotts Valley, CA, USA).

Total intake of SAA from base diet, powder and capsules is 50–60 mg/kg/days SAA and 15–25 mg/kg/days in the SAAhigh and SAAlow group, respectively. In human and animal studies, SAA restriction has usually involved ~ 80% reduction of SAA intake compared to the control group [7]. Notably, data on the normal range of SAA intakes in humans are limited. One paper by Nimni et al. estimated that Western and high-protein diets contain up to 50–70 mg/kg body weight/day of SAA in total, in comparison with ~ 20–30 mg/kg body weight/day in vegans/vegetarians [6]. The SAA restricted diet in our planned intervention will meet the WHO recommendations of SAA content of a minimum of 15 mg/kg body weight/day [27].

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