Table 1.

Neurocart test battery, EEG and related CNS domains

NeuroCart test	Targeted function and description of the test	Related CNS areas
Saccadic eye movement	Alertness, vigilance.58,63 The measurement of saccadic eye movements, specifically saccadic peak velocity, is highly sensitive for assessing sedation. Saccadic eye movements are captured using a moving dot displayed on a computer screen. Parameters such as latency, saccadic peak velocity, and inaccuracy of saccades were measured and analyzed.	Superior colliculus, substantia nigra, amygdala
Smooth pursuit	Visuomotor coordination.58,63 In the case of smooth pursuit eye movements, the target moved at frequencies ranging from 0.3 to 1.1 Hz, in increments of 0.1 Hz. The amplitude of target displacement corresponded to a rotation of 22.5 degrees for both sides of the eyeball. Four cycles were recorded for each stimulus frequency. The duration of time during which the eyes tracked the target in smooth pursuit was calculated for each frequency and expressed as a percentage of the stimulus duration. The average percentage of smooth pursuit across all stimulus frequencies was used as a parameter.	Midbrain
Adaptive tracking	Attention, visuomotor coordination.64 The adaptive tracking test was conducted using custom equipment and software, which was developed by Hobbs and Strutt, according to specifications of Borland and Nicholson. Analysis involved the average performance and standard deviation of scores over a 3.5-minute period, excluding the initial 0.5-minute run-in time. The task required the subject to track a moving circle on the screen by controlling a joystick to keep a dot within the target circle. Performance was deemed to be successful if the participant tracked the target at high speeds, while failure occurred when the velocity for tracking the target was low.	Neocortex, basal nuclei, brain stem, cerebellum
Body sway	Motor coordination, postural balance.65 Postural stability was assessed using a body sway meter, which measured body movements in a single plane to determine postural stability. The measurement of body sway was conducted using a pot string meter based on the Wright ataxiameter. A string was attached to the subject’s waist, capturing all body movements over a specific time period and quantifying them as millimeters of sway. The total measurement duration for body sway was two minutes.	Cerebellum, brain stem
Pupil size	Brain stem function. A digital camera was used to measure the size of subject’s pupils at different time points. One picture was taken from both eyes simultaneously, and the ratio between the pupil- and iris diameter was measured.	Brain stem, medulla oblongata
Visual verbal learning test (VVLT)	Episodic memory. Subjects that perform the VVLT were presented 30 words in three consecutive word trials, i.e. word learning test. Each trial ended with a free recall of the presented words (Immediate Recall) test to determine acquisition and consolidation of information). Approximately 2.5 h after start of the first trial, the subjects were asked to recall as many words as possible (Delayed Recall- this test measures active retrieval from long term memory). Immediately thereafter, the subjects underwent memory recognition test, which consisted of 15 presented words and 15 ‘distractors’ (Delayed Recognition- testing memory storage). A different word list was presented to each subject during the different occasions, in this way a subject was presented with a new word list each measurement.	Hippocampus
Stroop color-word interference task	A two-trial version of the color-word Stroop task was presented to the test subjects. In the first trial, 6 colored items were presented at random. The subjects were asked to respond as fast and as accurately as possible by pressing the keys 1, 2 or 3 on the number pad with the index finger, middle finger and ring finger of the dominant hand, corresponding with the correct answer. In the second trial, which appeared directly after the first trial, 34 color and word pairs were presented randomly to the subject, forming either congruent or incongruent matches. The subjects were again asked to respond as fast as possible by pressing the keys 1, 2 or 3 on the numpad, corresponding with the correct answer. Three colors were shown, which are green, red and blue. The colored items were presented in a random order.
Visual Analogue Scale (VAS) Bond and Lader	Alertness, mood, calmness.66 Participants were asked to indicate with a mouse click on the computer screen how they felt on sixteen visual analogue scales from which the following 3 main factors were calculated as described by: alertness (from nine scores), contentedness (often called mood; from five scores), and calmness (from two scores).	Cortex, prefrontal cortex
Visual Analogue Scale (VAS) Bowdle	Feeling high, internal and external perception.67–69 Potential subjective psychotomimetic (psychedelic) effects of antiglutamatergic agents can be evaluated using specific VAS. Bowdle Psychotomimetic Effects Scores consisted of thirteen 10 cm visual analogue lines ranging from 0 (‘not at all’) to 100 mm (‘extremely’), addressing various abnormal states of mind. The Bowdle VAS was administered electronically and took approximately 2 minutes to complete.	Cortex, prefrontal cortex, amygdala
Pharmaco-electroencephalography (pEEG)	CNS actions of pharmacological substances.70 Resting-state pEEG recordings were conducted on subjects with open and closed eyes for 5 minutes each. Subjects were instructed to avoid staring, head and eye movements, and suppress eye blinks while facing a featureless wall. pEEG was recorded using a 40-channel system, with electrode placement following the international 10–20 system. Ocular artifacts were detected using vertical and horizontal pEEG recordings. The recorded signals were processed by applying filters, calculating power spectrum density, and analyzing specific electrode sites of interest.	All brain regions

CNS, central nervous system; pEEG, pharmaco-electroencephalography; VAS, visual analogue scale; VVLT, Visual Verbal Learning Test.