Valuable information about cardiovascular system can be derived from the shape of aortic pulse wave being the result of reciprocal interaction between heart and vasculature. Pressure profiles in ascending aorta were obtained from peripheral waveforms recorded non-invasively (SphygmoCor, AtCor Medical, Australia) before, during and after hemodialysis sessions performed after 3-day and 2-day interdialytic intervals in 35 anuric, prevalent hemodialysis patients. Fluid status was assessed by Body Composition Monitor (Fresenius Medical Care, Bad Homburg, Germany) and online hematocrit monitoring device (CritLine, HemaMetrics, Utah). Systolic pressure and ejection duration decreased during dialysis. Augmentation index remained stable at 30 ± 13% throughout hemodialysis session despite the decrease of augmented pressure and pulse height. Subendocardial viability ratio (SEVR) determined after 3-day and 2-day interdialytic intervals increased during the sessions by 43.8 ± 26.6% and 26.1 ± 25.4%, respectively. Hemodialysis performed after 3-day and 2-day interdialytic periods reduced significantly overhydration by 2.4 ± 1.0 L and 1.8 ± 1.2 L and blood volume by 16.3 ± 9.7% and 13.7 ± 8.9%, respectively. Intradialytic increase of SEVR correlated with ultrafiltration rate (R = 0.39, p-value < 0.01), reduction in overhydration (R = -0.57, p-value < 0.001) and blood volume drop (R = -0.38, p-value < 0.01). The strong correlation between the decrease of overhydration during hemodialysis and increase in SEVR confirmed that careful fluid management is crucial for proper cardiac function. Hemodialysis affected cardiovascular system with the parameters derived from pulse-wave-analysis (systolic and augmented pressures, pulse height, ejection duration, SEVR) being significantly different at the end of dialysis from those before the session. Combination of pulse-wave-analysis with the monitoring of overhydration provides a new insight into the impact of hemodialysis on cardiovascular system.