Analytical Methods for Assessing Thiol Antioxidants in Biological Fluids: A Review.

Redox metabolism is an integral part of the glutathione system, encompassing reduced and oxidized glutathione, hydrogen peroxide, and associated enzymes. This core process orchestrates a network of thiol antioxidants like thioredoxins and peroxiredoxins, alongside critical thiol-containing proteins such as mercaptoalbumin. Modifications to thiol-containing proteins, including oxidation and glutathionylation, regulate cellular signaling influencing gene activities in inflammation and carcinogenesis. Analyzing thiol antioxidants, especially glutathione, in biological fluids offers insights into pathological conditions. This review discusses the analytical methods for biothiol determination, mainly in blood plasma. The study includes all key methodological aspects of spectroscopy, chromatography, electrochemistry, and mass spectrometry, highlighting their principles, benefits, limitations, and recent advancements that were not included in previously published reviews. Sample preparation and factors affecting thiol antioxidant measurements are discussed. The review reveals that the choice of analytical procedures should be based on the specific requirements of the research. Spectrophotometric methods are simple and cost-effective but may need more specificity. Chromatographic techniques have excellent separation capabilities but require longer analysis times. Electrochemical methods enable real-time monitoring but have disadvantages such as interference. Mass spectrometry-based approaches have high sensitivity and selectivity but require sophisticated instrumentation. Combining multiple techniques can provide comprehensive information on thiol antioxidant levels in biological fluids, enabling clearer insights into their roles in health and disease. This review covers the time span from 2010 to mid-2024, and the data were obtained from the SciFinder® (ACS), Google Scholar (Google), PubMed®, and ScienceDirect (Scopus) databases through a combination search approach using keywords.

Salivary Oxytocin and Antioxidative Response to Robotic Touch in Adults with Autism Spectrum Disorder.

Individuals with ASD are known to have a tendency to have tactile sensory processing issues that could be associated with their impairment as regards social communication. The alterations in tactile processing in autistic subjects are usually accompanied by hypersensitivity and other unpleasant emotions induced by tactile contact. In our study, we investigated the impact of the velocity and the force of a tactile stroke received impersonally by a custom-built robotic device. A total of 21 adults with ASD and 22 adults from a control group participated in our study. The participants' responses were assessed according to subjective scales, EEG changes, and the dynamics of saliva antioxidants and oxytocin. It was found that the oxytocin level was significantly lower in subjects with ASD but increased after tactile stimulation. However, contrary to expectations, the increase in the oxytocin level in the target group negatively correlated with the subjective pleasantness of tactile stimulation and was probably associated with a stress-induced effect. The basic levels of antioxidants did not differ between the TD and ASD groups; however, these had significantly increased in individuals with ASD by the end of the study. The EEG findings, which revealed enhanced antioxidant levels, contributed to the relief of the cognitive control during the study.

Effect of Low-Dose Line-Spectrum and Full-Spectrum UV on Major Humoral Components of Human Blood.

Ultraviolet blood irradiation (UVBI) is an alternative approach to the treatment of infectious diseases of various pathogeneses. Recently, UVBI has attracted particular interest as a new immunomodulatory method. Experimental studies available in the literature demonstrate the absence of precise mechanisms of the effect of ultraviolet radiation (UV) on blood. Here, we investigated the effect of UV radiation of line-spectrum mercury lamp (doses up to 500 mJ/cm2) traditionally used in UVBI on the major humoral blood components: albumin, globulins and uric acid. Preliminary data on the effect of various doses of UV radiation of full-spectrum flash xenon lamp (doses up to 136 mJ/cm2), a new promising source for UVBI, on the major blood plasma protein, albumin, are presented. The research methodology included spectrofluorimetric analysis of the oxidative modification of proteins and analysis of the antioxidant activity of humoral blood components by chemiluminometry. The effect of UV radiation on albumin caused its oxidative modification and, accordingly, an impairment of the transport properties of the protein. At the same time, UV-modified albumin and γ-globulins acquired pronounced antioxidant properties compared to native samples. Uric acid mixed with albumin did not protect the protein against UV-induced oxidation. The flash full-spectrum UV qualitatively had the same effect on albumin as line-spectrum UV did, but an order of magnitude lower doses were required to achieve comparable effects. The suggested protocol can be used for selecting a safe individual dose for UV therapy.

Composition of the Cytochrome c Complex with Cardiolipin by Thermal Lens Spectrometry.

Thermal lens spectrometry along with spectrophotometric titration were used to assess the composition of the complex of oxidized cytochrome c (ferricytochrome c) with 1,1',2,2'-tetraoleyl cardiolipin, which plays a key role in the initiation of apoptosis. Spectrophotometric titration was carried out for micromolar concentrations at which the complex is mainly insoluble, to assess the residual concentration in the solution and to estimate the solubility of the complex. Thermal lens spectrometry was used as a method of molecular absorption spectroscopy, which has two advantages over conventional optical transmission spectroscopy: the higher sensitivity of absorbance measurements and the possibility of studying the light absorption by chromophores and heat transfer in complex systems, such as living cells or tissues. Thermal lens measurements were carried out at nanomolar concentrations, where the complex is mainly in solution, i.e., under the conditions of its direct measurements. From the thermal lens measurements, the ratios of cytochrome c and cardiolipin in the complex were 50 at pH 7.4; 30 at pH 6.8; and 10 at pH 5.5, which fit well to the spectrophotometric data. The molecular solubility of the complex at pH 6.8-7.4 was estimated as 30 µmol/L.

Green and Sustainable Ultrasound-Assisted Anodic Electrochemical Preparation of Graphene Oxide Dispersions and Their Antioxidant Properties.

A fast method for preparing aqueous graphene oxide (GO) dispersions by electrochemical oxidation of a graphite anode without preliminary intercalation with oxidizing agents is proposed. Ultrasonic probing was used in the modulation mode of ultrasonic waves (work/rest) for more efficient graphite oxidation-exfoliation. It is shown that the 4/2 s mode of ultrasonic modulation is the most effective due to the probe material's low corrosion while maintaining the optimum synthesis temperature not exceeding 30-35 °C and achieving the best characteristics of the resulting product. Three cases of anodic oxidation of graphite to obtain graphene oxide were considered: (1) a combined cathode-anode compartment, (2) a split cathode-anode salt-bridged compartment, and (3) separated anode compartment with a 3.5 kDa dialysis membrane. It was determined that the approach to synthesis with a divided cathode-anode compartment makes it possible to obtain GO sheets with fewer defects compared to chemical methods or methods with a combined cathode-anode compartment and makes it possible to control the oxidation degree of the material (C:O ratio) by varying the current density. The prepared samples showed good stability for more than six months. The spectral and morphological characteristics were studied. Using chemiluminometry in the luminol/Co(II)/H2O2 system, the antioxidant properties concerning three key reactive oxygen species (H2O2, superoxide anion radical, and hydroxyl radical) were demonstrated. It was also shown that the prepared GO dispersions do not induce lipid and phospholipid peroxidation.

Antioxidant Potential of Aqueous Dispersions of Fullerenes C60, C70, and Gd@C82.

The antioxidant potential (capacity and activity) of aqueous fullerene dispersions (AFD) of non-functionalized C60, C70, and Gd@C82 endofullerene (in micromolar concentration range) was estimated based on chemiluminescence measurements of the model of luminol and generation of organic radicals by 2,2'-azobis(2-amidinopropane) dihydrochloride (ABAP). The antioxidant capacity was estimated by the TRAP method, from the concentration of half-suppression, and from the suppression area in the initial period. All three approaches agree and show that the antioxidant capacity of AFDs increased in the order Gd@C82 < C70 < C60. Mathematical modeling of the long-term kinetics data was used for antioxidant activity estimation. The effect of C60 and C70 is found to be quenching of the excited product of luminol with ABAP-generated radical and not an actual antioxidant effect; quenching constants differ insignificantly. Apart from quenching with a similar constant, the AFD of Gd@C82 exhibits actual antioxidant action. The antioxidant activity in Gd@C82 is 300-fold higher than quenching constants.

Effects of Aqueous Dispersions of C60, C70 and Gd@C82 Fullerenes on Genes Involved in Oxidative Stress and Anti-Inflammatory Pathways.

BACKGROUND: Fullerenes and metallofullerenes can be considered promising nanopharmaceuticals themselves and as a basis for chemical modification. As reactive oxygen species homeostasis plays a vital role in cells, the study of their effect on genes involved in oxidative stress and anti-inflammatory responses are of particular importance. METHODS: Human fetal lung fibroblasts were incubated with aqueous dispersions of C60, C70, and Gd@C82 in concentrations of 5 nM and 1.5 µM for 1, 3, 24, and 72 h. Cell viability, intracellular ROS, NOX4, NFκB, PRAR-γ, NRF2, heme oxygenase 1, and NAD(P)H quinone dehydrogenase 1 expression have been studied. RESULTS & CONCLUSION: The aqueous dispersions of C60, C70, and Gd@C82 fullerenes are active participants in reactive oxygen species (ROS) homeostasis. Low and high concentrations of aqueous fullerene dispersions (AFD) have similar effects. C70 was the most inert substance, C60 was the most active substance. All AFDs have both "prooxidant" and "antioxidant" effects but with a different balance. Gd@C82 was a substance with more pronounced antioxidant and anti-inflammatory properties, while C70 had more pronounced "prooxidant" properties.

Effect of Low-Dose Ionizing Radiation on the Expression of Mitochondria-Related Genes in Human Mesenchymal Stem Cells.

The concept of hormesis describes a phenomenon of adaptive response to low-dose ionizing radiation (LDIR). Similarly, the concept of mitohormesis states that the adaptive program in mitochondria is activated in response to minor stress effects. The mechanisms of hormesis effects are not clear, but it is assumed that they can be mediated by reactive oxygen species. Here, we studied effects of LDIR on mitochondria in mesenchymal stem cells. We have found that X-ray radiation at a dose of 10 cGy as well as oxidized fragments of cell-free DNA (cfDNA) at a concentration of 50 ng/mL resulted in an increased expression of a large number of genes regulating the function of the mitochondrial respiratory chain complexes in human mesenchymal stem cells (MSC). Several genes remained upregulated within hours after the exposure. Both X-ray radiation and oxidized cfDNA resulted in upregulation of FIS1 and MFN1 genes, which regulated fusion and fission of mitochondria, within 3-24 h after the exposure. Three hours after the exposure, the number of copies of mitochondrial DNA in cells had increased. These findings support the hypothesis that assumes oxidized cell-free DNA as a mediator of MSC response to low doses of radiation.

The Phosphonate Derivative of C60 Fullerene Induces Differentiation towards the Myogenic Lineage in Human Adipose-Derived Mesenchymal Stem Cells.

Inductors of myogenic stem cell differentiation attract attention, as they can be used to treat myodystrophies and post-traumatic injuries. Functionalization of fullerenes makes it possible to obtain water-soluble derivatives with targeted biochemical activity. This study examined the effects of the phosphonate C60 fullerene derivatives on the expression of myogenic transcription factors and myogenic differentiation of human mesenchymal stem cells (MSCs). Uptake of the phosphonate C60 fullerene derivatives in human MSCs, intracellular ROS visualization, superoxide scavenging potential, and the expression of myogenic, adipogenic, and osteogenic differentiation genes were studied. The prolonged MSC incubation (within 7-14 days) with the C60 pentaphoshonate potassium salt promoted their differentiation towards the myogenic lineage. The transcription factors and gene expressions determining myogenic differentiation (MYOD1, MYOG, MYF5, and MRF4) increased, while the expression of osteogenic differentiation factors (BMP2, BMP4, RUNX2, SPP1, and OCN) and adipogenic differentiation factors (CEBPB, LPL, and AP2 (FABP4)) was reduced or did not change. The stimulation of autophagy may be one of the factors contributing to the increased expression of myogenic differentiation genes in MSCs. Autophagy may be caused by intracellular alkalosis and/or short-term intracellular oxidative stress.

Perceived pleasantness of gentle touch in healthy individuals is related to salivary oxytocin response and EEG markers of arousal.

Affective touch plays an important role in human social bonding, affiliative behavior, and in general emotional well-being. A system of unmyelinated low-threshold mechanosensitive C-type afferents innervating hairy skin (C-tactile or CT system) is postulated to provide the neurophysiological background of affective touch perception. C-tactile afferents respond optimally to soft and slow strokes, and this response correlates positively with pleasure ratings of tactile stimuli. As gentle touch is consistently associated with oxytocin release further promoting prosocial behavior, it has been suggested that this effect is mediated by the response of C-tactile afferents. This study assesses a possible link between CT-optimal touch, its subjective pleasantness, EEG indices of cortical arousal, and peripheral oxytocin response. EEG was recorded in 28 healthy volunteers during resting state and tactile stimulation[gentle slow brush strokes on forearm (CT-targeted touch) and palm (non-CT-targeted touch)]. Saliva samples were collected before and after the touch stimulation. Oxytocin concentration increase was significantly associated with greater subjective ratings of CT-targeted touch but not of non-CT-targeted touch, and with lower peak alpha frequency values indicating decreased cortical arousal. The findings suggest that CT-targeted stimulation triggers oxytocin release but only when the touch is perceived at an individual level as having clearly positive affective salience. This corresponds to previous studies reporting that oxytocin response to touch can be related to different personality factors, and bears important implications for planning touch-based interventions in social and medical care.

Non-Functionalized Fullerenes and Endofullerenes in Aqueous Dispersions as Superoxide Scavengers.

Endohedral metal fullerene are potential nanopharmaceuticals for MRI; thus, it is important to study their effect on reactive oxygen species (ROS) homeostasis. Superoxide anion radical is one of the key ROS. The reactivity of aqueous dispersions of pristine (non-functionalized) fullerenes and Gd@C82 endofullerene have been studied with respect to superoxide in the xanthine/xanthine oxidase chemiluminescence system. It was found that C60 and C70 in aqueous dispersions react with superoxide as scavengers by a similar mechanism; differences in activity are determined by cluster parameters, primarily the concentration of available, acting molecules at the surface. Gd endofullerene is characterized by a significantly (one-and-a-half to two orders of magnitude) higher reactivity with respect to C60 and C70 and is likely to exhibit nanozyme (SOD-mimic) properties, which can be accounted for by the nonuniform distribution of electron density of the fullerene cage due to the presence of the endohedral atom; however, in the cell model, Gd@C82 showed the lowest activity compared to C60 and C70, which can be accounted for by its higher affinity for the lipid phase.

Antioxidant Potential of Antiviral Drug Umifenovir.

Free radical reactions play an important role in biological functions of living systems. The balance between oxidants and antioxidants is necessary for the normal homeostasis of cells and organisms. Experimental works demonstrate the role of oxidative stress that is caused by influenza virus as well as the toxic effects of some antiviral drugs. Therefore, antiviral drugs should be characterized by its pro- and antioxidant activity, because it can affect its therapeutic efficiency. The aim of the study was to quantify the antioxidant capacity and propose the mechanism of the antioxidant effect of the antiviral drug Umifenovir (Arbidol®). The kinetic chemiluminescence with the 2,2'-azobis (2-amidinopropane) dihydrochloride + luminol system was used to quantify the antioxidant capacity of Umifenovir relative to the standard compound Trolox. With computer simulation, the reaction scheme and rate constants were proposed. The antioxidant capacity of 0.9 µM Umifenovir (maximum concentration of Umifenovir in blood after oral administration of 200 mg) was as high as 1.65 ± 0.18 µM of Trolox. Thus, the total antioxidant capacity of Umifenovir is comparable to the antioxidant capacity of Trolox. Unlike Trolox, Umifenovir reacts with free radicals in two stages. For Trolox, the free radical scavenging rate constant was k = 2000 nM-1 min.-1, for Umifenovir k1 = 300 nM-1min.-1, k2 = 4 nM-1min.-1. Slower kinetics of Umifenovir provides the prolonged antioxidant effect when compared to Trolox. This phenomenon can make a serious contribution to the compensation of oxidative stress that is caused by a viral disease and the therapeutic effect of the drug.

Activity of NAD(P)H-Oxidoreductases in Ovarian Cancer.

Reactive oxygen species (ROS) play an important and controversial role in carcinogenesis. Microsomal redox chains containing NADH- and NADPH-dependent oxidoreductases are among the main sites of intracellular ROS synthesis, but their role in the oxidative balance has not been fully studied. Here, we studied the activity of cytochrome b5 reductase (CYB5R) and cytochrome P450 reductase (CYPOR) in ovarian cancer tissues and cells isolated from peritoneal fluid, along with the antioxidant capacity of peritoneal fluid. We used the developed a chemiluminescence assay based on stimulation with NADH and NADPH, which reflects the activity of CYB5R and CYPOR, respectively. The activity of CYB5R and CYPOR was significantly higher in moderately and poorly differentiated ovarian adenocarcinomas compared with well-differentiated adenocarcinomas and cystadenomas. For the chemotherapy-resistant tumors, the activity of tissue CYB5R and CYPOR was lower compared to the non-resistant tumors. In the peritoneal fluid, the antioxidant capacity significantly increased in this series, benign tumors < well-differentiated < moderately and poorly differentiated adenocarcinomas, so the antioxidant excess was observed for moderately and poorly differentiated adenocarcinomas. The antioxidant capacity of peritoneal fluid and the activity of CYB5R and CYPOR of cells isolated from peritoneal fluid were characterized by a direct moderate correlation for moderately and poorly differentiated adenocarcinomas. These results indicate the significant role of NAD(P)H oxidoreductases and the antioxidant potential of peritoneal fluid in cancer biochemistry. The parameters studied are useful for diagnostics and prognostics. The developed assay can be used to analyze CYB5R and CYPOR activity in other tissues and cells.

The Dynamics of Resting-State EEG and Salivary Trace Elements in Patients with Obsessive-Compulsive Disorder.

The study of salivary microelements and their neurophysiological and behavioral correlates in patients with obsessive-compulsive disorder (OCD) is a pressing issue in modern psychiatry, which, however, lacks adequate research at this time. In this study, we tested the dynamics of behavioral parameters, resting-state electroencephalogram (EEG), and salivary iron, copper, manganese, magnesium, and zinc in 30 healthy volunteers and 30 individuals with OCD before and after an emotional antisaccade task. The eye-movement data served as a measure of behavioral performance. Our research revealed consistently higher manganese concentrations in the OCD group compared to healthy volunteers associated with a higher EEG ratio of amplitude transformation and symptom severity. The dynamics of salivary microelements and resting-state EEG, possibly influenced by cognitive and emotional load during the anticsaccade task, differed between groups. In healthy volunteers, there was a decrease in salivary iron level with an increase in high-frequency power spectral density of EEG. The OCD group showed a decrease in salivary copper with an increased Hjorth mobility of EEG.

Acoustic and Subjective Basis of Emotional Perception in Comatose Patients: A Comparative Study.

Objective: : The acoustic stimulation in consciousness patients may improve the diagnosis assessment and the effectiveness of rehabilitation procedures. We aimed to investigate the event-related potential (ERP) response to emotional auditory stimuli in comatose patients. Methods: : We measured the nonlinear and linear electroencephalogram (EEG) features, prepared the acoustic analysis of stimuli parameters, and assessed the subjective emotional rates of stimuli characteristics. Results: : Patients with better outcomes had recognizable ERP responses and significant changes of the nonlinear EEG features to emotional sounds, unlike patients with worse outcomes. The response of comatose patients was attributed to acoustical features of emotional sounds, whereas the EEG response of healthy subjects was associated with their subjective feelings. The comatose patients demonstrated the variable EEG activity for neutral and emotional sounds. Conclusion: : Thus, the EEG reactivity followed the better outcome of comatose patients to emotional stimuli. The study assumed the substantial differences of emotional stimuli perception in the healthy and unconscious brain.

Albumin Retains Its Transport Function after Interaction with Cerium Dioxide Nanoparticles.

The interaction of inorganic nanomaterials with biological fluids containing proteins can lead not only to the formation of a protein corona and thereby to a change in the biological activity of nanoparticles but also to a significant effect on the structural and functional properties of the biomolecules themselves. This work studied the interaction of nanoscale CeO2, the most versatile nanozyme, with human serum albumin (HSA). Fourier transform infrared spectroscopy, MALDI-TOF mass spectrometry, UV-vis spectroscopy, and fluorescence spectroscopy confirmed the formation of HSA-CeO2 nanoparticle conjugates. Changes in protein conformation, which depend on the concentration of both citrate-stabilized CeO2 nanoparticles and pristine CeO2 nanoparticles, did not affect albumin drug-binding sites and, accordingly, did not impair the HSA transport function. The results obtained shed light on the biological consequences of the CeO2 nanoparticles' entrance into the body, which should be taken into account when engineering nanobiomaterials to increase their efficiency and reduce the side effects.

Salivary Antioxidant Capacity and Magnesium in Generalized Anxiety Disorder.

Generalized anxiety disorder (GAD) is a prevalent disorder. The search for biomarkers may contribute to new knowledge about molecular pathogenesis and treatment. Since oxidative stress and micronutrient imbalance play a key role in the development of mental disorders, we aimed to study salivary antioxidant capacity and magnesium in patients with GAD in an anxiety model of solving problems with increasing complexity. The study subgroup consisted of 15 patients with GAD, and 17 healthy volunteers of the same age made up the control subgroup. Participants took a test with six levels of difficulty, which included false feedback. In this test, the participants were asked to remember the colors of balloons and react when the color changed. The reaction time, the number of correct answers, as well as biochemical parameters such as the antioxidant capacity of saliva and salivary magnesium, were assessed. There was no difference in the results of the quest between the subgroups; however, anxious participants spent more time at the moment of experimental frustration due to incorrect feedback and additional negative psycho-emotional load. Antioxidant capacity did not differ between the subgroups both before and after the experimental session. Average antioxidant capacity also did not change significantly at the endpoint of the experiment. However, the endpoint antioxidant capacity correlated negatively with the reaction time in anxious patients in the second block (where the false feedback as a frustrating factor appeared). Magnesium was initially significantly higher in the group of anxious participants and decreased at the experiment endpoint; in healthy patients, there were no changes in salivary magnesium at the endpoint. In conclusion, the compensatory potential of oxidative metabolism and magnesium in patients with GAD was spent with additional psycho-emotional stress, in contrast to healthy individuals, but it was sufficient to avoid exhaustion during experimental frustrating exposure.

Effects of Aqueous Dispersions of C60, C70, and Gd@C82 Fullerenes on DNA Oxidative Damage/Repair and Apoptosis in Human Embryonic Lung Fibroblasts.

Fullerenes and metallofullerenes play an active role in homeostasis of reactive oxygen species and may cause oxidative damage to cells. As pristine fullerenes are a basis for derivatization, studying oxidative DNA damage/repair and apoptosis is important in terms of genotoxicity and cytotoxicity for their biomedical application. Aqueous dispersions of C60, C70, and Gd@C82 (5 nM and 1.5 µM) were cultured with human fetal lung fibroblasts for 1, 3, 24, and 72 h. Oxidative DNA damage/repair was assessed through concentration of 8-oxodG, double-strand breaks, and activation of BRCA1. Activity of apoptosis was assessed through the BCL2/BAX ratio. All three fullerenes caused oxidative modification of DNA at the early stages; C60 caused the most long-term damage, Gd@C82 caused the most short-term damage, and C70 caused "wave-like" dynamics. The dynamics of DNA repair correlated with the dynamics of oxidative damage, but Gd@C82 caused more prolonged activation of the repair system than C60 or C70. The oxidative toxicity of Gd@C82, is minor and the oxidative toxicity of C60 is mild and short-term, in contrast to C70. In relation to the studied effects, the fullerenes can be arranged in a safety row of Gd@C82 > C60 > C70.

High-Throughput Preparation of Uncontaminated Graphene-Oxide Aqueous Dispersions with Antioxidant Properties by Semi-Automated Diffusion Dialysis.

A semi-automated diffusion-dialysis purification procedure is proposed for the preparation of uncontaminated graphene oxide (GO) aqueous dispersions. The purification process is integrated with analytical-signal processing to control the purification degree online by several channels: oxidation-reduction potential, conductivity, and absorbance. This approach reduces the amounts of reagents for chemical treatment during dialysis. The total transition metal (Mn and Ti) content was reduced to a sub-ppb level (assessed by slurry nebulization in inductively coupled plasma optical atomic emission spectroscopy). Purified aqueous GO samples possess good stability for about a year with a zeta-potential of ca. -40 mV and a lateral size of ca. sub-µm. Purified GO samples showed increased antioxidant properties (up to five times compared to initial samples according to chemiluminometry by superoxide-radical (O2-) generated in situ from xanthine and xanthine oxidase with the lucigenin probe) and significantly decreased peroxidase-like activity (assessed by the H2O2-L-012 system).

Touch-induced emotional comfort results in an increase in the salivary antioxidant potential: A correlational study.

A pleasant touch reduces psychoemotional stress via the oxytocin mechanism due to its anti-inflammatory and antioxidant effects. Our research is aimed to reveal the correlations between the subjectively perceived pleasantness of touch, the antioxidant potential of saliva, and salivary oxytocin. A total of 56 healthy volunteers aged 18-38 years participated in the study. The control group consisted of 24 volunteers. The participants were subjected to tactile stimulation using a specially designed protocol. They ranked the touch pleasantness on a scale from 1 to 10. Heart rate variability and low-frequency/high-frequency ratios from the power spectral density of ECG were determined to assess psychoemotional relaxation. Salivary oxytocin and antioxidant capacity were quantified before and after the touch test. We found a significant increase in salivary antioxidant potential and oxytocin after pleasant tactile stimulation for the participants compared to the control group. The difference in antioxidant capacity values before and after the test positively correlated with mean pleasantness in the touch test (r = 0.57) and the difference in heart rate variability (r = 0.67); it negatively correlated with the difference in low-frequency/high-frequency ECG band ratio (r = -0.59). Oxytocin ratio positively correlated with the difference in antioxidant capacity values (r = 0.47). As a result of tactile stimulation, a significant increase in the antioxidant capacity of saliva and salivary oxytocin was found in the test group compared to the control group. These findings support further studies of the effects of pleasant touch on hormonal and oxidative metabolism.