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Background Diastolic dysfunction is an important factor in the development of heart failure with preserved ejection fraction (HFpEF). As the ejection fraction is preserved in HFpEF, the diagnosis of this disease with non-invasive methods is difficult. Purpose In this study, the relationship of BNP, NT-proBNP, Ghrelin, and echocardiographic 3D strain findings with diastolic dysfunction was investigated in patients undergoing left heart catheterization. Methods Our study is a cross-sectional study and included 78 patients in whom echocardiography was performed, and who underwent left heart catheterization based on relevant indications. The patient data recorded for evaluation included the findings from left heart catheterization, follow-up 3D echocardiography; and the levels of blood NT-proBNP, and Ghrelin. Results The rate of diastolic dysfunction was 42.3%. Longitudinal 2D and 3D mean strain as absolute values were observed to decrease more in patients with diastolic dysfunction. The median levels of BNP, NT-proBNP, and Ghrelin levels were higher in patients with diastolic dysfunction. The independent predictors of diastolic dysfunction were determined to be the left atrial volume index (LAVI) (OR=1.17; p=0.018), longitudinal 3D strain values (OR=1.88; p<0.001), NT-proBNP (OR=1.11; p=0.001), and Ghrelin (OR=1.40; p=0.001), respectively. Relationship Between LV EDP and LV Longitudinal Strain LV EDP 2D Strain 3D Strain r p r p r p BNP, pg/ml 0.429 <0.001* 0.115 0.316 0.178 0.118 NT-proBNP, pg/ml 0.484 <0.001* 0.155 0.177 0.186 0.104 Ghrelin, pg/ml 0.478 <0.001* 0.086 0.455 0.157 0.169 SolV DB – – 0.481 <0.001* 0.591 <0.001* dP/dT −0.389 <0.001* −0.283 0.012* −0.307 0.006* Negative dP/dT −0.747 <0.001* −0.337 0.003* −0.458 <0.001* 2D. % 0.481 <0.001* – – 0.852 <0.001* 3D. % 0.591 <0.001* 0.852 <0.001* – – If p value is less than 0.05 shows statistical significance. Measurement of longitudinal strain Conclusion In conclusion, our study found out that the reduced 3D strain absolute values and increased levels of NT-proBNP and Ghrelin biomarkers predicted diastolic dysfunction. If further large-scale studies prove the efficiency of these practical, they may not only allow for making a diagnosis of HFpEF more readily but may also eliminate the confusion in diagnostic algorithms. Acknowledgement/Funding None