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The two-step method requires the existence of causal relationships between the exposure, mediator, and outcome in pairs, otherwise, the mediator may not exhibit a true mediation effect [85]

It has been found that GLP-1R has glucose-dependent expression during osteogenic differentiation of MC3T3-E1 cells induced by bone morphogenetic protein-2 (BMP-2), and its expression increases with the increase of glucose concentration in the culture medium (Nuche-Berenguer et al., 2010)

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the study mentioned earlier noted a higher incidence of thiamine deficiency at 12 months in GLP-1 RA users compared to those not on these drugs, although its not extremely common