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Conclusion In healthy subjects, LSM increased after recent physical exercise, while alcohol consumption 24-72 h prior to examination did not have a significant impact. There was no clinically significant effect of breath hold on LSM. Our study supports present guidelines recommending rest prior to LSM, while indicating that breath hold may not be mandatory. Recent moderate alcohol exposure may affect LSM to a lesser extent than commonly believed.The bulk measurement of extracellular matrix (ECM) stiffness is commonly used in mechanobiology. However, past st