Plans for assessment and collection of outcomes {18a}
behavioral abnormalities, PVT, Exhaled breath, shock
SECONDARY, SHOCK
All data and biological specimens will be collected by trained study personnel. Standard operating procedures (SOP) for each study activity ensure consistent and high data quality. Participants will familiarize themselves with the equipment and performance tests (especially the driving simulator) during screening to prevent learning effects during the study sessions.For the primary outcome, unstimulated oral fluid will be collected using neutral Salivette® (Sarstedt, Sevelen, Germany) sampling devices. Samples will be stored at − 80 °C until trial conclusion and then extracted and analyzed in an untargeted metabolomics approach by ultra-performance liquid chromatography coupled to high-resolution mass spectrometry (UPLC-HRMS) equipment.For the secondary outcome measures, different procedures for data collection and assessment will be applied:Driving simulation will take place in a custom-modified full-chassis BMW i3 car, in which actuators replace shock absorbers to convey motion cues, such as road bumps, acceleration and braking. Placed in a 270-degree-projection and 360-degree-visibility setting, participants will drive through a fully immersive simulated environment provided by the driving simulation software (SILAB, “Szenariopaket Driver Fitness and Ability” Version 7.0; Würzburg Institute for Traffic Sciences GmbH, Veitshöchheim, Germany), offering a row of validated scenario modules. SILAB simultaneously records vehicle and driving parameters to evaluate, among others, standard deviation of the lateral position (SDLP). Standard driving errors are recorded automatically via Standardized Application for Fitness to Drive Evaluations (S.A.F.E.) software. Additionally, S.A.F.E. allows including observed qualified errors and scoring the driving performance (Fitness to Drive, FtD) [The PVT will be performed on laptops and its results will be calculated according to the validated protocol [The d2 Test of Attention will be performed on paper and its results will be calculated according to a standardized manual [Visual attention test will be performed using virtual reality glasses according to standardized procedure and assessment [Paper-based Karolinska Sleepiness Scale (Version A) questionnaires will be provided and assessed according to the validated procedure [Validated Stanford Sleepiness Scale questionnaires will be completed on laptops and follow a simple seven-scale choice [The EEG data, complemented by the polysomnographic (PSG) recording of electrooculogram (EOG), electromyogram (EMG), and electrocardiogram (ECG), will be collected with SIENNA® Ultimate amplifier systems (EMS Biomedical, Korneuburg, Austria) and analyzed with dedicated software for the visual scoring of sleep variables (e.g., alphatrace NeuroSpeed-PSG, alpha trace medical systems, Vienna, Austria). During the driving simulation, EEG data will be recorded using a mobile Philips ALICE 6 device (Philips AG Sleep and Respiratory Care, Zofingen, Switzerland). According to standard international criteria, sleep stages will be scored for each night in the laboratory, and validated sleep variables reflecting sleep quality (e.g., total sleep time, sleep latency, length of non-rapid-eye-movement (NREM) and rapid-eye-movement (REM) sleep stages, waking time after sleep onset, sleep efficiency) will be derived and compared between the conditions [Blinded and pre-instructed assessors will collect the behavioral abnormalities of participants after the driving simulation test on paper. They will rate orientation, coordination, speech, mood, appearance, reaction, and eye reactions to light according to standardized options.Unstimulated finger sweat will be collected using self-punched sampling units made of filter paper. Samples will be handled and analyzed analogously to oral fluid samples.Exhaled breath samples will be collected in specialized devices from SensAbues AB (Stockholm, Sweden) and BreathExplor® (Munkplast AB, Uppsala, Sweden). Dried blood spots will be collected on volumetric cards (Capitainer, Solna, Sweden). All samples will be handled and analyzed analogously to oral fluid samples.