Molecular actions of sex hormones in the brain and their potential treatment use in anxiety disorders.

Anxiety disorders are one of the most prevalent mood disorders that can lead to impaired quality of life. Current treatment of anxiety disorders has various adverse effects, safety concerns, or restricted efficacy; therefore, novel therapeutic targets need to be studied. Sex steroid hormones (SSHs) play a crucial role in the formation of brain structures, including regions of the limbic system and prefrontal cortex during perinatal development. In the brain, SSHs have activational and organizational effects mediated by either intracellular or transmembrane G-protein coupled receptors. During perinatal developmental periods, the physiological concentrations of SSHs lead to the normal development of the brain; however, the early hormonal dysregulation could result in various anxiety diorders later in life. Sex differences in the prevalence of anxiety disorders suggest that SSHs might be implicated in their development. In this review, we discuss preclinical and clinical studies regarding the role of dysregulated SSHs signaling during early brain development that modifies the risk for anxiety disorders in a sex-specific manner in adulthood. Moreover, our aim is to summarize potential molecular mechanisms by which the SSHs may affect anxiety disorders in preclinical research. Finally, the potential effects of SSHs in the treatment of anxiety disorders are discussed.

Sex steroid hormones in depressive disorders as a basis for new potential treatment strategies.

The sex steroid hormones (SSHs) such as testosterone, estradiol, progesterone, and their metabolites have important organizational and activational impacts on the brain during critical periods of brain development and in adulthood. A variety of slow and rapid mechanisms mediate both organizational and activational processes via intracellular or membrane receptors for SSHs. Physiological concentrations and distribution of SSHs in the brain result in normal brain development. Nevertheless, dysregulation of hormonal equilibrium may result in several mood disorders, including depressive disorders, later in adolescence or adulthood. Gender differences in cognitive abilities, emotions as well as the 2-3 times higher prevalence of depressive disorders in females, were already described. This implies that SSHs may play a role in the development of depressive disorders. In this review, we discuss preclinical and clinical studies linked to SSHs and development of depressive disorders. Our secondary aim includes a review of up-to-date knowledge about molecular mechanisms in the pathogenesis of depressive disorders. Understanding these molecular mechanisms might lead to significant treatment adjustments for patients with depressive disorders and to an amelioration of clinical outcomes for these patients. Nevertheless, the impact of SSHs on the brain in the context of the development of depressive disorders, progression, and treatment responsiveness is complex in nature, and depends upon several factors in concert such as gender, age, comorbidities, and general health conditions.

Testosterone, personality traits and aggressive driving among young male drivers.

INTRODUCTION: Psychological testing to examine potentially aggressive behaviour is a gold standard, but it is not sufficient. Testosterone might increase an aggressive behaviour. AIM: The aim of this study was to evaluate whether testosterone along with psychological assessment of fitness to drive could help to identify aggressive drivers. METHODS: Male participants (n=150) aged from 20 to 25, who possessed a driving license and drive at least 100 km per week, were evaluated in this study using an Inventory of traffic-relevant personality characteristics, the Sensation Seeking Scale and the Buss-Durkee Aggression Inventory. Saliva was collected for testosterone and cortisol measurements. The five binomial logistic models with dependent variables Caused an accident, Driving license taken away, Court trial, Intoxicated driving and Sporty self-report were tested in this study. RESULTS: The 'Intoxicated driving' model, was found to be statistically highly significant, explaining 48.8 % of the dependent variable's variance (χ2(16)=36.145, p<0.01). In this model with sensation seeking, actual testosterone and their interaction was highly significant and explained 20.4 % of intoxicated driving variability (χ2(3)=14.283, p<0.01). This was higher than sensation seeking scores only. CONCLUSION: To conclude, salivary testosterone might prove a biological marker that improves the identification of those with a high probability of aggressive driving or its subtypes (Tab. 3, Ref. 53).

Concentration of apelin inversely correlates with atrial fibrillation burden.

AIM: Asymptomatic atrial fibrillation (AF) detection and pulmonary veins isolation (PVI) outcome prediction remain challenging. Our aim was to study the association between apelin and paroxysmal AF in patients undergoing radiofrequency catheter PVI. METHODS: Sixty-three consecutive patients (55 ± 8years, 12 females) with paroxysmal AF without a structural heart disease and implanted ECG loop recorders undergoing PVI and healthy control group of 34 persons (41 ± 9.5years, 21 females) were included. Apelin plasmatic concentrations were measured before and three months after PVI. AF burden was continually assessed for three years. RESULTS: Apelin was significantly decreased in AF patients compared to the healthy controls (0.79 ± 0.09 vs 0.98 ± 0.06 ng/ml; p < 0.00001). Apelin plasmatic concentration of 0.89 ng/ml had 94 % specificity and 89 % sensitivity for AF prediction with the area under the curve (AUC) of 0.96. After propensity matching to sex, age and comorbidities, apelin concentration was significantly lower in AF group (0.78 ± 0.1 vs 0.99 ±0.06  ng/ml; p < 0.0001; AUC: 0.97). There was a significant inverse correlation between apelin concentration and AF burden both before and after PVI (Rho = ‒0.22; p = 0.05) and (Rho = ‒0.51; p = 0.006), respectively. There was no significant association between pre-PVI apelin and PVI long-term outcome. CONCLUSION: In patients without a structural heart disease apelin showed a significant specificity and sensitivity for AF prediction and inversely correlated with AF burden (Tab. 3, Fig. 3, Ref. 34).

Measurement of Urea in the Saliva of Healthy Mice - a Pilot Study.

Salivary urea is studied as a non-invasive alternative for screening and monitoring of renal diseases. Its high variability prevents a wider clinical use. Animal experiments are needed to identify factors affecting this marker. The aim of this study was to describe the inter-individual variability of salivary urea in healthy mice, establish reference intervals, and analyse the effects of sex, age and body weight. Plasma and saliva samples were obtained from 37 male and 41 female healthy adult CD1 mice aged 13-69 weeks (body weight 22-51 g). The reference interval for salivary urea in heathy mice based on our results is 2.7-8.4 mmol/l (CV = 23 %). Multivariate analysis did not show any significant effect of age, sex, or body weight. In addition, salivary urea did not correlate with its plasma concentrations. The high variability of the promising salivary marker of kidney function in healthy mice requires further research before its use to diagnose or monitor renal failure in animal models of kidney diseases. Other potential confounders should be analysed, including intra-individual and pre-analytical variability. In addition, a normalization factor such as total salivary proteins or salivation rate is likely needed.

Association of apelin and AF in patients with implanted loop recorders undergoing catheter ablation.

BACKGROUND: Previous studies showed an association between apelin and atrial fibrillation (AF). The aim of this study was to analyse the effect of pulmonary vein isolation (PVI) in patients with paroxysmal AF on plasmatic apelin concentrations. METHODS: Nine consecutive patients (aged from 43 to 69 years, 3 females and 6 males) with documented paroxysmal atrial fibrillation and implanted loop recorders (ILR) for continuous ECG monitoring were included in this study. All the patients underwent a radiofrequency catheter ablation with PVI. RESULTS: The plasmatic concentration of apelin increased after PVI. The average plasmatic concentration of apelin before PVI was 0.299 ng/ml (±0.16), 3 months after PVI 0.462 ng/ml (±0.10) and 9 months after PVI 0.565 ng/ml (±0.146). There was an increase in the concentration of apelin 3 months and 9 months after the PVI by 0.163 ng/ml (p=0.07) and by 0.266 ng/ml (p=0.01), respectively. The concentration of apelin inversely correlated with the AF burden (r=-0.44, p=0.03). CONCLUSIONS: Our study showed a significant increase in apelin levels after the reduction of AF burden via PVI and an inverse correlation with AF burden. Apelin might be a promising marker of AF (Tab. 2, Fig. 2, Ref. 28).

Localization dependent sensitivity of cerebral Na,K-ATPase to irradiation induced oxidative imbalance in rats.

Na,K-ATPase represents the key enzyme maintaining the ionic gradient across plasma membrane. It was documented that in directly irradiated organs the activity of this enzyme is decreased. The aim of present study was to clarify the remote effect of irradiation in mediastinal area on the activity of the Na,K-ATPase in selected brain regions in rats. Ionizing radiation in single dose 25 Gy induced alterations in oxidative status of blood plasma. Irradiation also decreased the activity of the Na,K-ATPase in cerebral cortex. Measurements of kinetic properties of the enzyme dependently on the concentration of energy substrate ATP or cofactor Na+ indicated that the lowered enzyme activity is probably a consequence of decreased number of active molecules of the enzyme, as suggested by lowered Vmax values (by 13 - 14%). Immunoblot analysis revealed that this effect is connected namely to decreased presence of α2 and α3 subunits (by 25% and 30% respectively). Considering the current concepts about involvement of the malfunction of α2 α3 subunits in development of primary brain dysfunctions, it may be hypothesized that the lowered functionality of those subunits of Na,K-ATPase may represent a predisposition to neurodegenerative disorders after irradiation. The observed effect seems to be localization dependent as the enzyme in cerebellum resisted to irradiation.

Metabolic and renal effects of dietary advanced glycation end products in pregnant rats - a pilot study.

Thermally processed food contains advanced glycation end products (AGEs) including N(epsilon)-(carboxymethyl)lysine (CML). Higher AGEs or circulating CML were shown to be associated with pregnancy complications such as preeclampsia and gestational diabetes. It is unclear whether this association is causal. The aim of our study was to analyze the effects of dietary CML and CML-containing thermally processed food on metabolism in pregnant rats. Animals were fed with standard or with AGE-rich diet from gestation day 1. Third group received standard diet and CML via gavage. On gestation day 18, blood pressure was measured, urine and blood were collected and the oral glucose tolerance test was performed. Plasma AGEs were slightly higher in pregnant rats fed with the AGE-rich diet (p=0.09). A non-significant trend towards higher CML in plasma was found in the CML group (p=0.06). No significant differences between groups were revealed in glucose metabolism or markers of renal functions like proteinuria and creatinine clearance. In conclusion, this study does not support the hypothesis that dietary AGEs such as CML might induce harmful metabolic changes or contribute to the pathogenesis of pregnancy complications. The short duration of the rodent gestation warrants further studies analyzing long-term effects of AGEs/CML in preconception nutrition.

Behavioral Changes During Development of Chronic Kidney Disease in Rats.

Decreased renal function due to chronic kidney disease (CKD) is associated with anxiety and cognitive decline. Although these mental disorders are often obvious in late stage renal disease patients, they might be unnoticeable or are neglected in early stages of the CKD development. Associations between renal and cognitive dysfunction have been indicated by studies performed mainly in patients undergoing dialysis, which itself represents a stress and decreased quality of life. However, experimental and causal studies are scarce. Our aim was to investigate dynamic changes in behavioral traits during the progression of CKD in an animal model. Thirty 12-week old male rats were used in this experiment. CKD was induced by a subtotal (5/6) nephrectomy. Two, 4, and 6 months after surgical induction of CKD, the open field, the light-dark box and the novel object recognition tests were conducted to assess the locomotor activity, anxiety-like behavior and the memory function of rats. Blood urea nitrogen (BUN), plasma concentration of creatinine (CREAT), albumin to creatinine ratio in urine (ACR) along with the renal histology were assessed to monitor the development and severity of CKD. In comparison to control rats, 5/6 nephrectomized rats had by 46-66% higher concentration of BUN during the whole follow-up period, as well as by 52% and by 167% higher CREAT and ACR, respectively, 6 months after surgery. Although the effect of time was observed in some behavioral parameters, nephrectomy did not significantly influence either locomotor activity, or anxiety-like behavior, or memory function of animals. Two and 4 months after surgery, animals moved shorter distance and spent less time in the center zone. However, the open-field ambulation returned back to the baseline level 6 months after CKD induction. Although nephrectomized rats displayed impaired kidney function as early as 2 months after surgery, no significant differences were found between the CKD and the control rats in any of the observed behaviors. Further studies are needed in order to evaluate whether behavioral abnormalities are related to severity of CKD or might be attributed to psychosocial aspect of end-stage renal disease and decreased quality of life in dialysis patients.

Markers of oxidative stress and antioxidant status in the plasma, urine and saliva of healthy mice.

Oxidative stress markers are usually measured in plasma, a stable environment for biomarkers. Blood collection is invasive, but the use of alternative biofluids is limited, due to high variability. In this study, we aimed to establish reference values for oxidative stress markers in plasma, urine and saliva of adult, healthy mice and to identify some sources of variability. Samples were obtained from 41 female and 37 male adult, healthy mice of the CD-1 strain, aged 95-480 days, weighing 21-55 grams. Reference ranges of TBARS (thiobarbituric acid reactive substances), AOPP (advanced oxidation protein products), fructosamine, GSH/GSSG (reduced and oxidized glutathione) ratio, TAC (total antioxidant capacity), and FRAP (ferric reducing antioxidant power) were measured in plasma and urine, and TBARS, GSH/GSSG ratio, TAC and FRAP in saliva, using standard spectrophotometric and fluorometric methods. Salivary GSH/GSSG and urinary AOPP were higher in females. Urinary fructosamine, GSH/GSSG and FRAP were higher in males. Urinary TAC and FRAP negatively correlated with age, and urinary GSH/GSSG positively correlated with weight. We determined that urine and saliva can be obtained non-invasively from mice, in sufficient amounts for reliable oxidative status assessment. Further studies are needed to uncover whether these biofluids reflect systemic oxidative status in diseases.

Putative Effects of Sex Hormones on Urinary Tract Infection.

Urinary tract infections affect mostly females. The infection and possible consequent ascent of bacteria is enhanced by various risk factors. Sex hormones regulate gene transcription implicated in immune cell development and maturation, in regulation of immune responses and immune signalling pathways. Limited knowledge is available; however, recent findings underline the importance of understanding the interactions between sex hormones and urinary tract infection to diminish the occurrence of complications related to this infection. This review summarizes and discusses the current knowledge on the correlation and impact of sex hormones on urinary tract infections.

Deoxyribonuclease partially ameliorates thioacetamide-induced hepatorenal injury.

Several recent studies have shown that liver injury is associated with the release of DNA from hepatocytes. This DNA stimulates innate immunity and induces sterile inflammation, exacerbating liver damage. Similar mechanisms have been described for acute renal injury. Deoxyribonuclease degrades cell-free DNA and can potentially prevent some of the induced tissue damage. This study analyzed the effects of thioacetamide-induced hepatorenal injury on plasma DNA in rats. Plasma DNA of both nuclear and mitochondrial origin was higher in thioacetamide-treated animals. Administration of deoxyribonuclease resulted in a mild, nonsignificant decrease in total plasma DNA and plasma DNA of mitochondrial origin but not of nuclear origin. This was accompanied by a decrease in bilirubin, creatinine, and blood urea nitrogen as markers of renal function. In conclusion, the study confirmed the hepatotoxic and nephrotoxic effect of thioacetamide. The associated increase in cell-free DNA seems to be involved in hepatorenal pathogenesis because treatment with deoxyribonuclease resulted in a partial prevention of hepatorenal injury. Further experiments will focus on the effects of long-term treatment with deoxyribonuclease in other clinically more relevant models. Clinical studies should test endogenous deoxyribonuclease activity as a potential risk determinant for kidney or liver failure.NEW & NOTEWORTHY Thioacetamide-induced hepatorenal injury resulted in higher plasma cell-free DNA. Deoxyribonuclease decreased average cell-free DNA of mitochondrial origin but not nuclear origin. Deoxyribonuclease partially prevented hepatorenal injury in rats.

Oxidative Stress and Antioxidants in the Diagnosis and Therapy of Periodontitis.

Oxidative stress has been implicated in the pathogenesis of numerous diseases. However, large interventional studies with antioxidants failed to show benefits in the prevention or treatment of cardiovascular diseases, cancer, or diabetes mellitus. Numerous clinical studies have confirmed the association of oxidative stress markers and periodontitis. Technical and biological variability is high for most of the analyzed markers and none of them seems to be optimal for routine clinical use. In a research setting, analysis of a palette of oxidative stress markers is needed to cover lipid peroxidation, protein oxidation, and the antioxidant status. The source of reactive oxygen species and their role in the pathogenesis of periodontitis remains unclear. Interventional experiments indicate that oxidative stress might be more than just a simple consequence of the inflammation. Small studies have confirmed that some antioxidants could have therapeutic value at least as an addition to the standard non-surgical treatment of periodontitis. A clear evidence for the efficiency of antioxidant treatment in large patient cohorts is lacking. Potentially, because lowering of oxidative stress markers might be a secondary effect of anti-inflammatory or antibacterial agents. As the field of research of oxidative stress in periodontitis gains attraction and the number of relevant published papers is increasing a systematic overview of the conducted observational and interventional studies is needed. This review summarizes the currently available literature linking oxidative stress and periodontitis and points toward the potential of adjuvant antioxidant treatment, especially in cases where standard treatment fails to improve the periodontal status.

The relation between oxidative stress biomarkers and atrial fibrillation after pulmonary veins isolation.

INTRODUCTION: The current paradigm claims a link between oxidative stress and atrial fibrillation. The aim of our research was to study a relation between the percentage of time spent in atrial fibrillation (AF burden) and concentrations of oxidative stress biomarkers, before and after pulmonary veins isolation (PVI). METHODOLOGY: We included 19 patients (mean age 55±10years, 4 females and 15 males) with implanted loop recorders undergoing PVI. Plasmatic concentrations of advanced glycation end-products (AGEs), fructosamine, advanced oxidation protein products and thiobarbituric-acid reacting substances (TBARS) were measured and AF burden was recorded immediately before and 3months after the PVI. AF burden was also recorded 9months after the PVI. RESULTS: Post procedural AGEs concentration significantly negatively correlated with AF burden after 3months (ρ=-0.63; p<0.01) and 9months (ρ=-0.5; p=0.04), respectively as well as TBARS concentration significantly negatively correlated with AF burden after 9months (ρ=-0.61; p=0.01). CONCLUSION: Our study showed AGEs and TBARS to be potential predictors for AF burden after the PVI. We suppose that the more oxidative stress after the PVI is provoked, the more fibrotic tissue is produced. That means a better electrical isolation of pulmonary veins and consequently a lower AF burden.

Variability of salivary markers of oxidative stress and antioxidant status in young healthy individuals.

OBJECTIVES: Salivary advanced glycation end-products (AGEs), advanced oxidation protein products (AOPP), total antioxidant capacity (TAC), and ferric reducing ability of saliva (FRAS) are increased in various diseases. Little data exist for these markers in the healthy population. The aim of this study was to assess the inter-individual and intra-individual variability of AGEs, AOPP, TAC, and FRAS in the saliva of young healthy individuals. METHODS: Unstimulated saliva samples were collected from 16 females and 18 males daily over a period of 30 days. Markers were measured using spectrophotometric and spectrofluorometric microplate-based methods. RESULTS: All salivary markers measured were significantly higher in men than in women (P < 0.05 for AGEs; P < 0.001 for AOPP, TAC, and FRAS). The inter-individual variability was approximately 60% for AGEs and AOPP and 30-40% for TAC and FRAS in both genders. The inter-individual variability of FRAS was higher in men vs. women (P < 0.01). Intra-individual variability ranged from 20% for TAC, to 30% for AGES and FRAS and 45% for AOPP. DISCUSSION: Intra-individual variability of salivary AGEs, AOPP, TAC, and FRAS indicates that their use is currently limited to large cohort studies. Identifying the underlying factors related to the high inter-individual and intra-individual variability is needed. Sex differences should be considered in future studies.

Salivary markers of oxidative stress in oral diseases.

Saliva is an interesting alternative diagnostic body fluid with several specific advantages over blood. These include non-invasive and easy collection and related possibility to do repeated sampling. One of the obstacles that hinders the wider use of saliva for diagnosis and monitoring of systemic diseases is its composition, which is affected by local oral status. However, this issue makes saliva very interesting for clinical biochemistry of oral diseases. Periodontitis, caries, oral precancerosis, and other local oral pathologies are associated with oxidative stress. Several markers of lipid peroxidation, protein oxidation and DNA damage induced by reactive oxygen species can be measured in saliva. Clinical studies have shown an association with oral pathologies at least for some of the established salivary markers of oxidative stress. This association is currently limited to the population level and none of the widely used markers can be applied for individual diagnostics. Oxidative stress seems to be of local oral origin, but it is currently unclear whether it is caused by an overproduction of reactive oxygen species due to inflammation or by the lack of antioxidants. Interventional studies, both, in experimental animals as well as humans indicate that antioxidant treatment could prevent or slow-down the progress of periodontitis. This makes the potential clinical use of salivary markers of oxidative stress even more attractive. This review summarizes basic information on the most commonly used salivary markers of oxidative damage, antioxidant status, and carbonyl stress and the studies analyzing these markers in patients with caries or periodontitis.

Anthocyanin-Rich Diet in Chemically Induced Colitis in Mice.

The aetiology of inflammatory bowel diseases is unclear, but oxidative stress plays a key role in the pathogenesis. Anthocyanins--plant polyphenols--were shown to have antioxidant and anti-inflammatory properties. The aim of this study was to investigate the potential protective effects of anthocyanins on the oxidative status in mice with chemically induced colitis. Adult male mice were randomly divided into a control group drinking tap water and a colitis group drinking 1% dextran sulphate sodium solution. Animals had ad libitum access to a control wheat-based diet or food based on wheat producing anthocyanins. Bodyweight and stool consistency were monitored daily for 14 days. At the end of the experiment, colon length was measured and tissue samples were collected for the assessment of histology and oxidative status. Mice with colitis had lower body weight, higher stool score and shorter colon than control mice. Anthocyanins had neither an effect on stool consistency, nor on bodyweight loss and colon length. In the colon, liver and plasma, analysis of oxidative stress markers and antioxidant status revealed no significant differences between the groups. Food made from wheat producing anthocyanins did not protect mice from the consequences of chemically induced colitis. The measured biomarkers do not confirm the role of oxidative stress in this model of colitis. Further optimization of the anthocyanin-rich food might be needed before further experiments are conducted.

The renal effects of prenatal testosterone in rats.

PURPOSE: Previous studies have shown that prenatal testosterone affects the development of not only reproductive organs but also the brain and even glucose metabolism. Whether prenatal testosterone influences the kidney development is largely unknown. We analyzed whether testosterone modulation during prenatal development would affect renal function and the number of nephrons in adult offspring. MATERIALS AND METHODS: Pregnant rats were treated with olive oil, testosterone (2 mg/kg), the androgen receptor blocker flutamide (5 mg/kg) or testosterone plus flutamide via daily intramuscular injections from gestation day 14 until delivery. Renal histology and functional parameters were assessed in male and female adult offspring. Macerated kidneys were used for nephron counting. RESULTS: Prenatal testosterone administration increased proteinuria in male rats by 256%. A similar 134% effect in female rats was not statistically significant. This effect was prevented when flutamide was co-administered. In male rats prenatal testosterone increased blood urea nitrogen. In female rats flutamide increased creatinine clearance. In male rats prenatal testosterone and flutamide led to higher and lower, respectively, interstitial collagen deposition in adulthood. CONCLUSIONS: Prenatal testosterone induces proteinuria in adulthood. This effect is mediated via androgen receptor. Additional effects seem to be sex specific. Further studies should focus on the timing and dosing of testosterone as well as the applicability to human development.

BSE-associated polymorphisms in the prion protein gene: an investigation.

The aim of this study was to determine the frequency of the 12-bp and 23-bp indel polymorphisms in the prion protein gene (PRNP) in cattle and to investigate the association between these frequencies and the occurrence of bovine spongiform encephalopathy (BSE). There was no significant difference in the 12-bp indel frequency between the BSE animals and control group. For the 23-bp indel, the BSE animals had a significantly lower + + (insins) genotype frequency and + allele frequency compared with the control animals. The - - / - - genotype frequency in the BSE animals was not significantly higher when compared with the control animals. One - allele increased the risk of BSE by a factor of 1.55 (i.e. by 55%) for the 12-bp indel and by a factor of 2.10 for the 23-bp indel. When both indels are considered, one - allele increased the risk of BSE by a factor of 1.54.

Salivary markers of oxidative stress in patients with obstructive sleep apnea treated with continuous positive airway pressure.

PURPOSE: Obstructive sleep apnea syndrome (OSAS) is characterized by elevated oxidative stress. Measurement of oxidative stress in saliva seems to be promising in long-term treatment monitoring of OSAS patients. In this study, our aim was to investigate whether short-term continuous positive airway pressure (CPAP) treatment would influence oxidative stress in saliva. METHODS: Patients with diagnosed OSAS (16 women, 28 men) underwent polysomnography during the first night and CPAP treatment during the second night. Saliva samples were taken in the evening and morning on both days. Markers of oxidative stress and antioxidant status were analyzed in saliva. RESULTS: Evening concentrations of the salivary thiobarbituric acid reacting substances (p < 0.001), advanced glycation end-products (p < 0.001), and advanced oxidation protein products (p < 0.01) were significantly lower than morning values during the diagnostic night. However, salivary concentrations of none of the oxidative stress markers were significantly influenced by the CPAP treatment. No changes in salivary antioxidant status after CPAP therapy were found. CONCLUSION: Salivary markers of oxidative stress and antioxidant status do not change significantly after one night treatment with CPAP. On the contrary, after 1 month with CPAP therapy, reduced markers of oxidative stress were reported. Therefore, the future studies should be focused on finding the optimal sampling frequency to clarify the potential of saliva for the monitoring of OSAS treatment.