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44 patients with CAD underwent endothelial function testing. Segments with >10% vasoconstriction after acetylcholine (ACh) were defined as having severe EDFx. WSS, calculated using computational fluid dynamics, was defined as low (< 1 Pa), intermediate (1–2.5 Pa) or high (> 2.5 Pa). In 4,510 coronary segments, higher proportion of segments with low WSS had severe EDFx (71%) compared to intermediate WSS (22%) or high WSS (23%) (p < 0.001). Segments with low WSS demonstrated greater vasoconstriction in response to ACh than intermediate or high WSS segments (p < 0.001). Low WSS (OR: 9.14, p < 0.001) independently predicted severe EDFx.