Pupillary alterations in virtual humans induce neurophysiological responses within an observer. Technological advances have enabled rapid developments in artificial intelligence (AI), from verbal systems, to visual AI interfaces with the ability to express, and respond to emotional states of a user. Visual AI interfaces are able to change their physical parameters, such as pupil diameter. Pupillary changes can alter heart rate, however, effects on heart rate variability (HRV) are unknown. HRV, is an autonomic, non-conscious parameter which monitors sympathetic and parasympathetic nervous system (PNS) activity. N = 34 male participants aged between 19–33 were subjected to a number of conditions such as pupil dilation, constriction and blushing. The present research is the first to investigate the effects of virtual human interactions on human HRV. Outcomes of this study were obtained using eye tracking and HRV measurements. Pupil dilation relative to constriction presented in the female virtual partner induced a significant right pupillary diameter increase (p = 0.041) in human observers. Additionally, female virtual partner pupil constriction relative to dilation induced a significant increase in participants’ PNS HRV response (p = 0.036). These findings indicate the ability of a female virtual interaction partner to modulate parasympathetic autonomic functioning in young healthy male humans. This allows first insights into the effects of interacting with virtual AI interaction partners, on human autonomic functioning, and may aid development of future virtual humans, and their implementation into relevant clinical settings.
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