2.1 Participants
Twenty females have received remuneration for participation in this experiment (Age: 39.8 ± 11.9 years old), after providing written informed consent. They reported normal or corrected-to-normal vision and no history of psychiatric and neurological disorders. This study was performed in accordance with the principles in the Declaration of Helsinki and approved by the Ethical committee at the Shiseido Global Innovation Center (Approval number: C02121).
2.2 Stimuli and procedure
The participants comfortably sat on a chair and were asked to perform 4 behavioural tasks according to the instructions that appeared on the PC screen in front of them (Fig. 1). The first task was the "photo" task where the participants watched a movie. An experimenter appeared in the movie and pretended to take a picture of the participant, who was asked to smile. The second was the "smiling" task. The participants made their biggest smile that they can immediately after a beep was heard from the PC. Participants were instructed how to make the biggest smile (pull the mouth corner back maximally) and the participants decided whether to open their mouth or not themselves before the experiment. During the experiment, they replicated this type of smile. The third was the "speech” task. The participants spoke the designated 4 sentences (from an ATR 503 phonetically balanced sentences) on the screen. The fourth task was the "free talk" task. The participants talked about 2 themes (“What I'm addicted to these days” and “What I have enjoyed recently”) as long as they could up to a maximum of 30 seconds. They were also asked to pretend to talk with a person familiar to them in this task.
Photo and smiling tasks were both conducted twice (once at the beginning and once at the end of the experiment) for each mask condition. All tasks were implemented with or without a surgical mask on their face (Fig. 1). The order of mask condition (with or without mask) was counterbalanced for the repetition (once at the beginning and once at the end of the experiment) and across participants. After the experiment was started, the experimenter left the participants alone in the experimental room as an anti-coronavirus infection risk countermeasure and let them initiate the start of the experiment by themselves which they did by pushing the spacebar of the PC. During this time, the participant was recorded on video. Detailed procedures of the behavioural tasks and timing of wearing and taking off the mask were instructed by the in-house developed Python 3.7.3 (Python software foundation, https://www.python.org/) program made with the Psychopy library 19.
2.3 Data acquisition and analysis
Three active bipolar Ag electrodes (AP-C140-020, Miyuki Giken Co., Ltd., Tokyo, Japan) were attached on the face to acquire the electromyography (EMG) from orbicularis oculi, zygomaticus major of the left facial sites, and depressor anguli oris of the right facial sites. The detailed positioning was decided in accordance with published guidelines 20. Reference and Grand electrodes (AP-C151-015 and MA-C004-015 with general disposal electrode, respectively) were both attached on the forehead and covered with a hairband. To lower the electrode conductance, we applied Ten20 conductive paste (Weaver and Company, Aurora, CO, USA) on the electrode tips and attached a pierced double-sided tape between the body of the electrode and skin. They were also covered with elastic tape.
EMG signals were recorded by a 32-ch physiological signal recording device (Polymate V AP5148, Miyuki Giken Co., Ltd., Tokyo, Japan) with a sampling rate of 1000 Hz. 50-Hz noise in the EMG was excluded by a notch filter (48–52 Hz). Then the spiky outlier data over 200 mV was removed. The denoised signal was rectified and integrated with each 20-ms time window 20 to monitor its time series (Fig. 2–5). The preprocessed EMG signals were divided into epochs of each behavioural task based on the audio signals (beep for the smiling task and voice onset for the photo, speech, and free talk tasks). Due to the self-proceeding experimental procedure, the epochs where the participants failed the procedures were manually checked on the recorded video and excluded from further analyses. This process was done by a rater who took no further role in the data analysis. These failures included failure to comply with the task (e.g. due to failure to hear instructions), misspeaking or misreading. In the photo and smiling tasks, if both trials were rated as successful, the signals from the two trials were averaged. If only one trial was rated as a success and one as a fail, only the successful trial was used in further analysis. The EMG signal was averaged during the tasks and compared between conditions with or without the mask with Wilcoxon signed-rank test with a Bonferroni correction (N = 12 for tasks and measured muscles).