Salivary Cytokine Profile in Patients with Ischemic Stroke. Can Salivary TNF-α be a Non-invasive Biomarker of Cognitive Impairment in Stroke Cases?

Salivary Cytokine Profile in Patients with Ischemic Stroke. Can Salivary TNF-α be a Non-invasive Biomarker of Cognitive Impairment in Stroke Cases? | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Research Article Salivary Cytokine Profile in Patients with Ischemic Stroke. Can Salivary TNF-α be a Non-invasive Biomarker of Cognitive Impairment in Stroke Cases? Mateusz Maciejczyk, Kacper Mil, Piotr Gerreth, Katarzyna Hojan, and 2 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-494834/v1 This work is licensed under a CC BY 4.0 License Status: Under Review Version 1 posted 9 You are reading this latest preprint version Abstract Inflammation plays a crucial role in stroke pathogenesis. Thus, it is not surprising that cytokines, chemokines, and growth factors have been advocated in stroke diagnostics. Our study is the first to evaluate the salivary cytokine profile in patients with ischemic stroke. Twenty-five patients with subacute ischemic stroke and an age-, sex-, and oral hygiene status-matched control group were enrolled in the study. The number of patients was set a priori based on our previous experiment (α = 0.05, test power = 0.9). Salivary concentrations of tumor necrosis factor α (TNF-α), interleukin 6 (IL-6), and interleukin 10 (IL-10) were assessed using an ELISA method. We showed that salivary TNF-α and IL-6 were significantly higher, whereas IL-10 content was statistically lower in both non-stimulated (NWS) and stimulated (SWS) whole saliva of ischemic stroke patients. However, evaluation of cytokines in NWS rather than in SWS may be of greater diagnostic value. Of particular note is salivary TNF-α, which may indicate cognitive/physical impairment in post-stroke individuals. This parameter distinguishes stroke patients from healthy controls and correlates with cognitive dysfunction and severity of functional impairment. It also differentiates (with high sensitivity and specificity) stroke patients with normal cognition from mild to moderate dementia patients. Saliva may be an alternative to blood for assessing cytokines in stroke patients, although further studies on a larger patient population are needed. Cardiac & Cardiovascular Systems Cognitive Neuroscience stroke saliva inflammation cytokines salivary biomarkers salivary diagnostics Figures Figure 1 Introduction Stroke is a clinical syndrome characterized by the sudden onset of focal and sometimes generalized brain damage with symptoms lasting longer than 24 hours 1 . According to the World Health Organization (WHO), stroke is the third leading cause of death in the adult population 2 . It is estimated that 5.5 million people die annually from this disease, accounting for 10% of all deaths 3 . Stroke is also the leading cause of long-term functional impairment and disability in adults 4 . There are two types of stroke: 10–15% are hemorrhagic strokes, and the rest are ischemic strokes 5 . Inflammation plays a crucial role in stroke pathogenesis 2 , 6 , 7 . Acute cerebral ischemia induces cellular-molecular mechanisms involving both the central nervous system (CNS) and the vascular system. In response to ischemic brain injury, acute inflammatory reactions are induced, mediated by various cytokines, chemokines, and growth factors 8 . Among them, a unique role is attributed to TNF-α (tumor necrosis factor α), which is the first mediator synthesized in response to acute cerebral ischemia 6 . Under these conditions, other pro-inflammatory cytokines are activated (e.g., interleukin-6 (IL-6) and interleukin-8 (IL-8)). TNF-α also upregulates the expression of matrix metalloproteinases (MMPs), which increases endothelial cell permeability and results in blood-brain barrier (BBB) dysfunction 9 , 10 . Pro-inflammatory mediators exert direct damaging effects on neurons and astrocytes and promote prostaglandin release and leukocyte migration 2 , 6 , 7 . It has been shown that IL-6 and TNF-α stimulate the synthesis and secretion of several chemotactic factors, which activate and attract peripheral blood leukocytes (neutrophils and monocytes) to the site of stroke lesions 9 , 11 . Therefore, adhesion molecules' expression on the endothelial surface is increased, which potentiates leukocyte penetration into the ischemic brain area. Simultaneously, the secretion of interleukin-10 (IL-10) increases, inhibiting the inflammatory response 12 . Stroke diagnosis is based on physical examination and imaging studies (CT) 13 . Despite the precise definition of stroke, up to 20–25% of diagnoses may be conditions that mimic stroke 14 . Availability of the latest diagnostic techniques (CT perfusion, MRI) is also often limited 15 . Therefore, it is not surprising that new diagnostic methods are still being sought 16 . In particular, biochemical biomarkers are being searched extensively to facilitate early diagnosis of stroke or to establish further patient’s prognosis 17 , 18 . Given the critical role of inflammation in stroke pathogenesis, the use of cytokines and chemokines in laboratory diagnostics has been increasingly advocated 16 , 19 . Of particular note is the evaluation of TNF-α and IL-6, whose serum levels increase in the acute phase of stroke as well as correlate with its cerebrospinal fluid (CSF) levels and stroke focus size 20 – 23 . The high diagnostic utility also seems to be demonstrated by IL-10, whose levels decrease in patients with ischemic stroke and indicate the predominance of pro-inflammatory processes in the early stroke phase 12 , 24 . Nowadays, the use of biological materials collected in a non-invasive manner, without special equipment and medical personnel, is gaining interest in laboratory diagnostics 25 , 26 . This reduces the patient's anxiety associated with the examination, promotes more frequent monitoring of health condition, and thus diagnoses the disorder at an early stage 27 . Saliva is an interesting alternative to blood or CSF that are commonly used in laboratory medicine 25 , 28 . Saliva is a secretion of major and minor salivary glands containing water (94–99%) and numerous proteins, lipids, carbohydrates, hormones, immunoglobulins, cytokines, growth factors, and vitamins 29 – 31 . Its composition is therefore very similar to blood plasma. Indeed, the level of many compounds in saliva correlates with their content in blood/CSF, indicating a solid diagnostic potential 25,28,32−34 . Although salivary cytokines are used to diagnose many systemic diseases 35 – 39 , there is no data on salivary inflammatory mediators' diagnostic utility in stroke individuals. Considering the continuous increase in stroke morbidity/mortality and the lack of non-invasive biomarkers of the disease, our study aimed to evaluate TNF-α, IL-6, and IL-10 levels in the non-stimulated (NWS) and stimulated (SWS) whole saliva of stroke patients. Material And Methods Study population The study was carried out from June to August 2019 in a health center (Bonifraterskie Centrum Zdrowia) in Piaski–Marysin (Piaski, Poland). In this center, patients with various disorders are hospitalized, including the individuals after the cerebral stroke that arrive from different country provinces. At the time of the research, there were 315 patients in the neurorehabilitation unit following different incidents, including vascular brain damage, brain injury, spinal cord injury, surgically treated patients with a brain tumor, sclerosis multiplex, myelopathy, and polyneuropathy. It was determined that 218 (65.71%) patients were stroke survivors, and the individuals were admitted to the neurorehabilitation ward immediately from the hospital, directly after the acute phase cessation, in a subacute phase of the stroke. A medical doctor assessed each patient at the beginning of the stay at the unit, and afterward, he/she was suggested to comprehensive individual and similar rehabilitation. Most of the patients were able to cooperate, understand instructions, and communicate. In addition, the majority of the individuals followed the same diet, including a baseline diet for most patients or a diet for people with diabetes mellitus. All the meals were prepared in the hospital and were distributed to the patients at the same time daily. Decisively, 97 (30.79%) individuals with other diseases than stroke were excluded from the study, and 25 (11.47% of stroke survivors; 7.94% of all individuals rehabilitated at the health center fully completed the examination, saliva sampling, and were considered in the analysis (Table 1 ). The other stroke patients had to be excluded from the study because of the following reasons: 80 (36.70%) patients refused to participate in the study; 42 (19.27%) people were uncooperative since they could not give conscious written informed consent for participation in the research and/or entirely communicate; 28 (12.84%) patients did not report for a sampling of saliva and dental examination, although they gave informed consent and were reminded up to 4 times; 11 (5.04%) subjects withdrew from the research after sampling of non-stimulated saliva due to physiological or psychological fatigue; 5 (2.29%) people were not able to gather the saliva due to issues concerning understanding the procedure because of cognitive and language deficits, and 2 (0.92%) patients were transferred to another hospital because of deterioration of general health. Essential information on the general condition and health status of patients were obtained from individuals' files and contained: gender, age, the medication used, time since diagnosis of cerebral stroke, and medical history (Table 1 ). Table 1 General characteristics of patients. Control n = 25 Stroke n = 25 p value Mean ± SD 95%CI Mean ± SD 95%CI Sex (male/female) 12/13 12/13 > 0.9999 Age 63.52 ± 8.545 59.99–67.05 63.52 ± 8.545 59.99–67.05 > 0.9999 Cognitive and physical functional status ACE III 97.24 ± 1.422 96.65–97.83 69.8 ± 22.97 60.32–79.28 < 0.0001 BI 20 ± 0 20–20 10.92 ± 3.662 9.408–12.43 < 0.0001 BBS 55.52 ± 0.5099 55.31–55.73 31 ± 16.27 24.28–37.72 < 0.0001 FIM 125.3 ± 0.6904 125–125.6 86.88 ± 30.31 74.37–99.39 0.9999 diabetes n (%) 4 (16) 4 (16) > 0.9999 epilepsy n (%) 4 (16) 3 (12) > 0.9999 arteriosclerosis n (%) 7 (28) 8 (32) > 0.9999 limb thrombosis n (%) 2 (8) 2 (8) > 0.9999 atrial fibrillation n (%) 3 (12) 3 (12) > 0.9999 Drugs 0.9999 ≥ 5 drugs/day n (%) 15 (60) 16 (64) > 0.9999 Abbreviations: ACE III – Addenbrooke’s Cognitive Examination; BI – Barthel Index; BBS – Berg Balance Scale; FIM – functional independence measure; differences statistically significant at: * p < 0.05; ** p < 0.005; *** p < 0.0005; **** p < 0.0001. For measurement of the functional status of the individuals the following scales were used: Addenbrooke’s Cognitive Examination III (ACE III) for differentiation between patients without and with cognitive impairment 40 . The Barthel Index (BI) to measure performance in activities of daily living (ADL) 41 . The Berg Balance Scale (BBS) to determine an individual's ability or inability to safely balance during a series of predetermined tasks 42 . The functional independence measure (FIM) for exploration of the patient's psychological, physical, and social functioning 43 . One experienced medical doctor, a neurorehabilitation specialist, qualified all the patients for the examination according to study criteria (Table 2 ). Table 2 Inclusion and exclusion criteria for the subjects in the study group. Inclusion Criteria Exclusion Criteria age of consent (> 18 years) nonage (< 18 years) consciousness unconsciousness giving of written and informed consent for saliva sampling and oral examination lack of written and informed consent for saliva sampling and oral examination from patient good general condition poor general condition confirmed cerebral infarction or cerebral hemorrhage based on CT and magnetic resonance imaging (MRI) unconfirmed cerebral infarction or cerebral hemorrhage based on CT and magnetic resonance imaging (MRI) adequate capacity to follow instructions, i.e., understanding how to perform the procedures, ability to collect a saliva sample, and being able to answer questions during the examination inadequate capacity to follow instructions, i.e., not understanding how to perform the procedures, inability to collect a saliva sample, and being unable to answer questions during the examination recovery from the acute phase of ischemic or hemorrhagic stroke in all brain areas no recovery from the acute phase of ischemic or hemorrhagic stroke in all brain areas no stroke recurrence during the subacute phase stroke recurrence during the subacute phase good cooperation insufficient cooperation due to cognitive and/or language deficits ischemic stroke treated without thrombectomy or thrombolysis ischemic stroke treated with thrombectomy or thrombolysis capability to collect a saliva sample incapability to collect a saliva sample no legal guardianship legal guardianship patients without psychiatric or cognitive disorders patients with psychiatric or cognitive disorders no autoimmune disease (rheumatoid arthritis, systemic lupus erythematosus) autoimmune disease (rheumatoid arthritis, systemic lupus erythematosus) no lung disease (chronic obstructive pulmonary disease) or cardiovascular disease (angina or uncontrolled hypertension) lung disease (chronic obstructive pulmonary disease) or cardiovascular disease (angina or uncontrolled hypertension) good general condition heart failure above NYHA II no intake of dietary supplements and vitamins for the last three months intake of dietary supplements and vitamins for the last three months non-smokers smokers women/men without malnutrition patients with malnutrition, i.e., having body mass index 10% during the previous three months the first admission to cure stroke unit was more than 5–6 (to 10) hours from the onset of the early neurological symptoms Control group The control group included 25 generally healthy patients who reported for oral examination to the Department of Restorative Dentistry of the Medical University of Bialystok (Bialystok, Poland) from March to September 2017. Controls were similar to the stroke patients regarding age and gender, oral hygiene, dentition, and periodontium status (Table 1 ). All participants were provided with medical clearance by clinicians before involvement in the research. Individuals from the control group followed a regular, balanced diet (which was not restricted), and they were given standard physical activity recommendations. Sampling of saliva The research material was total mixed non-stimulated saliva (NWS) and stimulated saliva (SWS), and both samples were collected via spitting. Saliva sampling was done in the health center during summertime, i.e., between June and September, to keep similar weather conditions outside. Before the procedure, individuals were instructed not to intake any solid food and/or liquid, other than clean water, at least two hours before saliva sampling. All individuals were in the subacute phase of the stroke. The study and control group individuals had not taken any medication eight hours before sampling of saliva 44 , 45 . The oral bioliquid was gathered, from subjects that had restrained from intensive physical activity for the preceding twelve hours, i.e., between 7:30 a.m. and 9:00 a.m. The patients were also indicated not to carry out any oral hygiene practices (i.e., mouth rinsing, teeth brushing, gum chewing, etc.) 45 , 46 . The saliva was collected after a five‐minute adaptation to the environment in a separate, private room. The oral cavity was rinsed two times with distilled water at room temperature before gathering saliva to avoid possible impurity from other sources. During the procedure, subjects were seated in an adjustable chair, individually adapted to each individual's height, with the head slightly bent downwards and resting in a convenient position, and they tried to limit their lips and face movements 45 , 46 . The samples of saliva that were stored during the first minute were ejected. Saliva was accumulated into a sterile Falcon tube placed in a container with ice. The non‐stimulated saliva was collected for 10 minutes and stimulated saliva was gathered in the same manner for 5 minutes to avoid the patients' physiological and/or psychological fatigue. The secretion of this oral bioliquid was stimulated by ten µL of 2% citric acid applied to the tongue's central part every 30 seconds. Afterward, the volume of saliva was measured using a calibrated pipette with an accuracy of 0.1 mL 47 . The minute flow of NWS and SWS was measured by dividing the saliva volume by the time essential for its secretion (mL/min). Forthwith, after collection of saliva, it was centrifuged (+ 4°C, 20 min, 3000 × g; MPW 351, MPW Med. Instruments, Warsaw, Poland). To protect againts sample oxidation, butylated hydroxytoluene (BHT, Sigma‐Aldrich, Saint Louis, MO, USA) was added into the acquired supernatants in the amount of 10 µL 0.5 M BHT in acetonitrile (ACN)/1 mL of saliva 48 . The samples of NWS and SWS were frozen at − 80°C and stored for no more than three months for further investigation. Oral examination The dentition was evaluated in a separate room shortly after saliva sampling. According to the World Health Organization criteria, the individual's check-up was carried out using a plain mouth mirror and a dental probe in artificial lighting 49 , 50 . The dentist performed the dental examination in front of the chair where the patient was seated and with the individual's head resting against the wall. Each accessible tooth surface was assessed and scored as generally healthy, decayed (DT), extracted due to the carious process (MT), or filled because of caries (FT). The data collected were used to calculate the DMFT index, that expresses dental caries experience, and it is the sum of the following components: DT, MT, and FT. Additionally, the prevalence of dental caries was determined as a percentage of patients with DMFT > 0. Gingival index (GI) and Plaque Index (PlI) were evaluated on the teeth 16, 12, 24, 36, 32, 44 using four-degree scales (from 0 to 3) 51 . The oral examination was performed by two dentists (P.G. and K.G.), subsequently after sampling of non-stimulated (NWS) and stimulated (SWS) whole saliva, and after previous training and calibration by an experienced dental specialist (A.Z.). The inter‐examiner and intra‐examiner agreement for DMFT, PlI and GI was evaluated by another dental examination in 10 patients with a κ > 0.94. Patient’s consent and Bioethical Issues All individuals, i.e., stroke patients and healthy controls, voluntarily took part in the study, and they were provided with comprehensive information concerning its purpose, procedures, risks, and benefits. Full written consent was obtained from all participants in accordance with the Declaration of Helsinki for a sampling of saliva and dental examination. Before the research, it was approved by the Bioethics Committee of the Poznan University of Medical Sciences (resolutions 59/19 and 890/19). Salivary cytokine profile Salivary concentrations of tumor necrosis factor α (TNF-α), interleukin 6 (IL-6), and interleukin 10 (IL-10) were assessed using commercial enzyme-linked immunosorbent assay (ELISA) kits according to the manufacturer’s instructions (EIAab, Wuhan, China). Briefly, specific antibodies suitably labeled with an enzyme were added to the antigen-coated plate. Depending on the amount of antigen, antibodies specifically bound to the antigen and unbound antibodies were eluted. After adding the substrate/chromophore, the enzyme-catalyzed the reaction, and the colored reaction product was measured colorimetrically at 405 nm using Mindray MR-96 Microplate Reader. All determinations were performed in duplicate samples. Salivary total protein content was determined in duplicate samples by the bicinchoninic acid (BCA) method (Pierce BCA Protein Assay Kit; Rockford, IL, USA). All results were presented as concentration (pg/mL), salivary output (pg/min), and specific content (mg/mg protein). The TNF-α /IL-10 and IL-6/IL-10 ratios were also calculated. Statistical data analysis Statistical analysis was carried out using GraphPad Prism 8 for macOS (GraphPad Software, La Jolla, CA, USA). Due to the fact that most of the results were characterized by a normal distribution (Kolmogorov-Smirnov test), the t-student test was used to compare two groups. If the distribution was not normal, the Mann-Whitney U test was used. ANOVA analysis of variance with Tukey's post hoc test was used to compare the 3 groups, and p values were calculated with correction for multiple comparisons. Correlations between salivary biomarkers and clinical data were performed using the Pearson correlation coefficient. The results were presented as mean ± SD and 95% confidence interval (95% CI). The assumed statistical significance was p < 0.05. Cohen Kappa (online calculator) was used to establish the inter-and intracorporeal agreement between the evaluated dental indicators. The diagnostic usefulness of salivary cytokines was determined using ROC (Receiver Operating Curve) analysis by evaluating the area under the curve (AUC). The sensitivity and specificity and the cut-off point, which was characterized by the highest sensitivity and specificity, were given. The number of patients was set a priori based on our previous study. For this purpose, an online sample size calculator (ClinCalc) was used. The level of significance was set at 0.05, and the power of the study was 0.9. Variables used for sample size calculation were concentration of salivary IL-6 and IL-10. The minimum number of patients was 21 (for one group). Results Table 3 Salivary gland function and dental characteristics of the stroke and healthy controls. Control n = 25 Stroke n = 25 p value Mean ± SD 95%CI Mean ± SD 95%CI NWS Flow rate (mL/min) 0.33 ± 0.08461 0.2951–0.3649 0.394 ± 0.2459 0.2925–0.4955 0.2245 Total protein (µg/mL) 1220 ± 174 1148–1292 1141 ± 281.5 1025–1257 0.2391 SWS Flow rate (mL/min) 0.9112 ± 0.2629 0.8027-1.02 0.5796 ± 0.2454 0.4783–0.6809 < 0.0001 Total protein (µg/mL) 1310 ± 172.3 1239–1382 947 ± 244.8 846–1048 < 0.0001 DMFT 24.4 ± 7.848 21.16–27.64 23.08 ± 7.989 19.78–26.38 0.5584 GI 0.8136 ± 0.8085 0.4799–1.147 0.72 ± 0.7294 0.4189–1.021 0.6693 PlI 1.254 ± 0.9743 0.8514–1.656 1.234 ± 0.9953 0.8228–1.644 0.9431 Abbreviations: NWS – non-stimulated whole saliva; SWS – stimulated whole saliva; DMFT index - a sum of decayed teeth (DT), teeth missing due to carious process (MT), and teeth filled because of caries (FT); GI – Gingival Index; PlI – Plaque Index. Salivary gland function The secretion of non-stimulated saliva did not differ significantly between the study and control groups. Stimulated saliva secretion was significantly decreased (↓36%) in the stroke population than the controls (Table 3 ). The concentration of total protein in non-stimulated saliva did not differ significantly between healthy controls and stroke individuals. The total protein concentration in stimulated saliva of the stroke population was significantly lower (↓28%) than in the control group (Table 3 ). Oral health status DMFT, GI, and PlI were not significantly different between the study and control group (Table 3 ). Salivary cytokines Non-stimulated saliva of stroke patients was characterized by significantly higher concentration (↑135%), output (↑196%), and specific amount (↑170%) of TNF-α. Similarly, these parameters were increased regarding IL-6 (respectively: ↑85%, ↑124%, 103%). On the other hand, the concentration (↓47%), output (↓78%), and specific amount (↓42%) of IL-10 decreased. As a result of these changes, the TNF-α/IL-10 ratio (↑507%) and IL-6/IL-10 ratio (↑378%) increased markedly. The results of measurements in stimulated saliva were comparable to those in non-stimulated saliva. The concentration (↑68%) and specific amount (↑147%) of TNF-α were significantly higher in patients with stroke than in control. Mentioned parameters were also higher in relation to IL-6 (concentration: ↑62%, specific amount: ↑57%). The output of TNF-α and IL-6 remained statistically insignificant. IL-10 concentration, output, and specific amount decreased, similarly to NWS (respectively: ↓65%, ↑36%, ↑48%). In consequence, TNF-α/IL-10 (↑611%) and IL-6/IL-10 (↑568%) ratios increased greatly (Table 4 ). Correlations Statistically significant correlations were summed up in Table 5 . The positive correlations were revealed only in non-stimulated saliva between IL-6 concentration and ADL, and IL-6 specific amount and ADL. The strong negative correlations in non-stimulated saliva were observed between TNF-α concentration and BBS, TNF-α concentration and ACE III or TNF-α concentration and ADL. Moreover, IL-6 specific amount and PlI and IL-6 output and FIM correlate negatively in unstimulated saliva. The negative correlation between TNF-α output and BBS was noted in either stimulated or non-stimulated saliva. In stimulated saliva, TNF-α output correlated negatively with BBS, ADL and ACE III. On the other hand, IL-6 output was observed to correlate negatively with BBS and ADL. Results of all correlations were presented in the supplementary material. ROC analysis The results of ROC analysis were presented in Tables 6 and 7 . The majority of analyzed cytokines significantly differentiated the patients with ischemic stroke from healthy controls. What is worth mentioning, many of given parameters in stimulated saliva, were characterized by AUC = 1, sensitivity and specificity reaching up to 100%, e.g. TNF-α/IL-10 ratio, IL-6/IL-10 ratio, TNF-α specific amount, and IL-6 concentration. In non-stimulated saliva, TNF-α/IL-10 and IL-6/IL-10 ratios distinguished themselves with high AUC and both great sensitivity and specificity. TNF-α diagnostic significance Salivary TNF correlates remarkably negatively with cognitive function in the ACE III scale. Therefore, we examined how this parameter differentiates stroke patients according to cognitive impairment. For this purpose, stroke patients were divided into 3 groups: normal cognitive function (100 − 89 in the ACE III scale), mild dementia (88 − 61 in the ACE III scale), and moderate dementia (< 61 in the ACE III scale) 52 , 53 . Salivary TNF-α differentiates (with high sensitivity and specificity) stroke patients with normal cognitive function from patients with mild to moderate dementia (Fig. 1 ). Table 4 Salivary cytokines in stroke patients and healthy controls. Salivary biomarker NWS SWS Control n = 25 Stroke n = 25 p value Control n = 25 Stroke n = 25 p value Mean ± SD 95%CI Mean ± SD 95%CI Mean ± SD 95%CI Mean ± SD 95%CI TNF-α concentration (pg/mL) 11.72±3.503 10.27-13.16 27.93±6.021 25.44-30.41 <0.0001 4.02±0.6272 3.761-4.279 6.776±0.8391 6.43-7.122 <0.0001 output (pg/min) 3.731±1.181 3.244-4.218 11.07±7.472 7.989-14.16 <0.0001 3.649±1.189 3.158-4.14 3.947±1.768 3.217-4.677 0.4884 specific amount (ng/mg protein) 9.846±3.591 8.364-11.33 26.63±11.21 22-31.25 <0.0001 3.106±0.5689 2.871-3.341 7.702±2.53 6.658-8.746 <0.0001 IL-6 concentration (pg/mL) 15.81±3.715 14.28-17.34 29.27±9.684 25.28-33.27 <0.0001 6.996±0.8605 6.64-7.351 11.3±1.065 10.86-11.74 <0.0001 output (pg/min) 5.213±1.863 4.443-5.982 11.68±8.251 8.27-15.08 0.0004 6.37±2.004 5.543-7.197 6.487±2.655 5.391-7.583 0.8614 specific amount (ng/mg protein) 13.27±3.674 11.75-14.79 26.89±9.536 22.95-30.82 <0.0001 5.449±1.094 4.997-5.9 12.8±3.838 11.21-14.38 <0.0001 IL-10 concentration (pg/mL) 3.934±0.7284 3.634-4.235 2.078±0.8456 1.728-2.427 <0.000 0.5012±0.07068 0.4721-0.5304 0.1753±0.085 0.1402-0.2104 <0.0001 output (pg/min) 0.4502±0.1308 0.3962-0.5042 0.09977±0.05173 0.07842-0.1211 0.0031 1.287±0.3666 1.136-1.438 0.8238±0.6476 0.5564-1.091 <0.0001 specific amount (ng/mg protein) 3.303±0.8475 2.953-3.653 1.927±0.9295 1.543-2.31 <0.0001 0.3881±0.07196 0.3584-0.4178 0.2015±0.1105 0.1559-0.2471 <0.0001 TNF-α/IL-10 ratio 3.119±1.285 2.588-3.649 18.96±17.54 11.71-26.2 <0.0001 8.186±1.776 7.452-8.919 58.24±56.44 34.94-81.54 <0.0001 IL-6/IL-10 ratio 4.156±1.28 3.627-4.684 19.88±22.59 10.56-29.21 0.0011 14.33±3.247 12.99-15.67 95.8±86.71 60.01-131.6 <0.0001 Abbreviations: TNF-α - tumor necrosis factor α; IL-6 – interleukin 6; IL-10 – interleukin 10 Table 5 Statistically significant correlations in patients with ischemic stroke. Pair of variables r p value NWS TNF-α concentration and BBS -0.612 0.001 TNF-α concentration and BI -0.789 < 0.001 TNF-α concentration and ACE III -0.832 < 0.001 TNF-α output and BBS -0.398 0.045 IL-6 concentration and BI 0.464 0.02 IL-6 output and FIM -0.41 0.042 IL-6 specific amount and PlI -0.481 0.015 IL-6 specific amount and BI 0.418 0.037 SWS TNF-α output and BBS -0.474 0.017 TNF-α output and BI -0.761 < 0.001 TNF-α output and ACE III -0.801 < 0.001 IL-6 output and BBS -0.512 0.009 IL-6 output and BI -0.796 < 0.001 IL-6 output and ACE III -0.754 < 0.001 Abbreviations: NWS – non-stimulated whole saliva; SWS – stimulated whole saliva; TNF-α - tumor necrosis factor α; IL-6 – interleukin 6; PLI – Plaque Index; ACE III – Addenbrooke’s Cognitive Examination; FIM – functional independence measure; BBS – Berg Balance Scale; Table 6 Receiver operating characteristics of salivary cytokines in non-stimulated saliva of stroke patients and healthy controls. Salivary biomarker NWS AUC 95% CI P-value Cut off Sensitivity% 95% CI Specificity% 95% CI TNF-α concentration (pg/mL) 0.992 0.9760–1.000 17.46 96 80.46–99.79 96 80.46–99.79 output (pg/min) 0.8544 0.7470–0.9618 4.727 76 56.57–88.50 80 60.87–91.14 specific amount (ng/mg protein) 0.9712 0.9340–1.000 15.61 92 75.03–98.58 92 75.03–98.58 IL-6 concentration (pg/mL) 0.8976 0.7944–1.000 22.08 84 65.35–93.60 96 80.46–99.79 output (pg/min) 0.7376 0.5897–0.8855 0.004 > 5.602 76 56.57–88.50 72 52.42–85.72 specific amount (ng/mg protein) 0.9104 0.8168–1.000 18.03 84 65.35–93.60 92 75.03–98.58 IL-10 concentration (pg/mL) 0.9568 0.9078–1.000 < 0.0001 < 3.014 88 70.04–95.83 92 75.03–98.58 output (pg/min) 0.776 0.6400–0.9120 0.0008 < 1.140 80 60.87–91.14 68 48.41–82.79 specific amount (ng/mg protein) 0.8848 0.7883–0.9813 < 0.0001 < 2.173 72 52.42–85.72 96 80.46–99.79 TNF-α/IL-10 0.9984 0.9931–1.000 5.577 100 86.68–100.0 96 80.46–99.79 IL-6/IL-10 0.9792 0.9426–1.000 7.625 92 75.03–98.58 100 86.68–100.0 Abbreviations: TNF-α - tumor necrosis factor α; IL-6 – interleukin 6; IL-10 – interleukin 10. Table 7 Receiver operating characteristics of salivary cytokines in stimulated saliva of stroke patients and healthy controls. Salivary biomarker SWS AUC 95% CI P-value Cut off Sensitivity% 95% CI Specificity% 95% CI TNF-α concentration (pg/mL) 0.9984 0.9931 to 1.000 4.941 100 86.68–100.0 96 80.46–99.79 output (pg/min) 0.5616 0.3950 to 0.7282 0.4551 > 4.211 52 33.50–69.97 76 56.57–88.50 specific amount (ng/mg protein) 1 1.000–1.000 4.249 100 86.68–100.0 100 86.68–100.0 IL-6 concentration (pg/mL) 1 1.000–1.000 8.986 100 86.68–100.0 100 86.68–100.0 output (pg/min) 0.536 0.3704–0.7016 0.6624 > 6.640 60 40.74–76.60 60 40.74–76.60 specific amount (ng/mg protein) 0.9968 0.9882–1.000 8.366 96 80.46–99.79 100 86.68–100.0 IL-10 concentration (pg/mL) 0.9984 0.9931–1.000 < 0.0001 < 0.3165 96 80.46–99.79 100 86.68–100.0 output (pg/min) 1 1.000–1.000 < 0.0001 < 0.2250 100 86.68–100.0 100 86.68–100.0 specific amount (ng/mg protein) 0.9136 0.8353–0.9919 < 0.0001 < 0.2665 76 56.57–88.50 96 80.46–99.79 TNF-α/IL-10 1 1.000–1.000 17.01 100 86.68–100.0 100 86.68–100.0 IL-6/IL-10 1 1.000–1.000 24.60 100 86.68–100.0 100 86.68–100.0 Abbreviations: TNF-α - tumor necrosis factor α; IL-6 – interleukin 6; IL-10 – interleukin 10. Discussion Our study is the first to evaluate the salivary cytokine profile in patients with ischemic stroke. We have demonstrated that assessment of TNF-α, IL-6, and IL-10 levels in both NWS and SWS significantly differentiates stroke patients from healthy controls. However, particular attention should be paid to salivary TNF-α, which may be a non-invasive biomarker of cognitive function/physical status in stroke subjects. In recent years, increasing attention has been given to the potential use of saliva in laboratory diagnostics. This biomaterial allows for repeated, non-invasive, and stress-free sampling, which is especially important in disabled and elderly patients 54 . Saliva is also a non-infectious and relatively durable bioliquid 29 , 55 . The concentration of many biomarkers in saliva highly correlates with their blood/CSF levels, allowing saliva to be used in the diagnosis of various systemic diseases 56 – 59 . Indeed, saliva can be an alternative to blood for the determination of enzymes, hormones, immunoglobulins, and redox biomarkers 60 , 61 . Saliva has also been shown to be valuable in assessing cytokines, chemokines, pro-inflammatory enzymes, and growth factors 62 – 64 . Although salivary cytokines' diagnostic utility has been demonstrated in metabolic and neurodegenerative diseases 35 – 39 , there are no data on ischemic stroke patients. We showed that salivary TNF-α and IL-6 were significantly higher, whereas IL-10 content was statistically lower in ischemic stroke patients than healthy controls. TNF-α/IL-10 and IL-6/IL-10 ratio were also significantly higher in NWS and SWS of stroke patients, indicating an intensification of the inflammatory process and a limited anti-inflammatory role of IL-10. As we have shown by ROC analysis, salivary biomarkers are also characterized by a high diagnostic utility in stroke diagnostics. Salivary TNF-α, IL-6, and IL-10 as well as TNF-α/IL-10 and IL-6/IL-10 ratio significantly distinguish stroke patients from controls with high sensitivity and specificity. This fact should not be surprising because inflammatory and immune factors have been indicated as potential stroke biomarkers 65 , 66 . Indeed, in stroke patients, activation of CNS cells (mainly microglia and astrocytes) and increased expression of blood cytokines, interleukins, chemokines, adhesion molecules, and pro-inflammatory enzymes is observed 67 . Interestingly, the severity of inflammation correlates with brain infarct size and neurological deficits in CT and physical examination 68−70 . However, cytokines' diagnostic utility has only been demonstrated in plasma/serum and CSF of ischemic stroke patients 70−72 . Our study is the first to show the utility of saliva to determine cytokine levels in stroke subjects. This raises hopes for non-invasive, painless, and non-stressful stroke diagnostics. It is believed that cytokines synthesized in the stroke focus pass through the damaged BBB into the blood and then into saliva by passive transport (diffusion or ultrafiltration) or by a specific transporter (facilitated diffusion or active transport) 73 , 74 . The BBB has a prominent role in the development of the inflammatory response by increasing the cytokine circulation between the CSF and the bloodstream 9 . Unfortunately, in our research, we did not assess the cytokine content in blood, so further studies are needed to evaluate stroke patients' saliva-blood relationship. Nevertheless, it can be speculated that, as in other CNS diseases, salivary cytokine concentrations reflect their plasma/CSF levels 25 , 28 , 75 . In our patients, the content of pro-inflammatory TNF-α and IL-6 was higher, and anti-inflammatory IL-10 was lower in both NWS and SWS of stroke patients. However, it is important to note that saliva's cytokine profile depends on the type of salivary gland involved in saliva secretion. Indeed, up to 70% of non-stimulated saliva is produced in the submandibular glands, making the NWS composition very similar to blood plasma. During stimulation, the parotid gland's contribution to saliva formation increases 76 . Stimulated saliva is characterized by significantly higher concentrations of Na + , HCO 3 − and Cl − ions and lower contents of K + and phosphate ions and mucins compared to non-stimulated saliva 77 . Cytokine production in salivary gland cells also increases, making the composition of SWS more different from that of plasma 78 . Therefore, evaluation of cytokines in NWS rather than in SWS may be of greater diagnostic value. In our study, among all assessed cytokines, tumor necrosis factor α deserves special attention. Salivary TNF-α differentiates stroke patients from controls with high sensitivity and specificity and correlates with cognitive dysfunction in the ACE III scale and severity of functional impairment in the BBS and ADL scale. Therefore, in the next step, we examined how this parameter distinguishes stroke patients according to cognitive impairment. For this purpose, stroke subjects were divided into three groups: normal cognition (100 − 89 in the ACE III scale), mild dementia (88 − 61 in the ACE III scale), and moderate dementia (< 61 in the ACE III scale) 52 , 53 . Salivary TNF-α differentiates (with high sensitivity and specificity) stroke patients with normal cognitive function from patients with mild to moderate dementia (Fig. 1 ). Although further studies in a larger patient population are needed, TNF-α may be a potential biomarker of cognitive impairment/functional status in stroke cases. As shown in previous studies, elevated TNF-α serum level was noted already in the first few hours after the ischemic stroke 10 , 16 . Serum TNF-α also correlates with brain infarct volume and severity of neurological symptoms as well as reflects the degree of patients' functional disability 79 , 80 . Indeed, TNF-α has a unique role in stroke pathogenesis. Primarily, it is the first inflammatory mediator produced in the ischemic area 6 , 80 . Secondly, it stimulates the synthesis/secretion of chemotactic factors that stimulate peripheral blood leukocytes and activates other cytokines like IL-1, IL-6, and interferon-gamma (IFN-γ) 10 . TNF-α also enhances MMPs activity 81 , thereby increasing endothelial cell permeability and BBB dysfunction 82 . Our study is the first to demonstrate that salivary TNF-α can be a potential biomarker of stroke patient’s functioning. Interestingly, the level of TNF-α in NWS correlates more strongly with the cognitive and physical functional status of stroke patients. Thus, non-stimulated saliva has a better diagnostic potential than SWS. Despite the unquestionable advantages, the use of saliva in diagnostics also has some drawbacks. The biomarker concentration in saliva may be highly dependent on the degree of salivary gland secretion 83 . Because salivary hypofunction may occur in stroke patients 84 , 85 , in our study, the concentration of inflammatory biomarkers was standardized to total protein content and salivary flow rate. Nevertheless, both the concentration (pg/mL), salivary output (pg/min), and specific content (per mg protein) of TNF-α and IL-6 were significantly higher, whereas IL-10 was statistically lower in NWS and SWS of stroke patients compared to controls. Importantly, we did not observe a relationship between salivary gland function and the assessed cytokines. This suggests a negligible effect of salivary hypofunction on salivary TNF-α, IL-6, and IL-10 levels. Nevertheless, aging, medications, diet supplements, smoking, as well as oral and systemic diseases can also modify the cytokine profile of saliva 86 – 88 . At this time, it is important to emphasize the careful selection of the control group in the presented study. Control individuals are matched not only by age and sex to the study group but also by other clinical features, including concomitant diseases, medications, oral hygiene status, etc. This significantly reduces the impact of confounding factors on the obtained results. In summary, stroke patients have increased salivary secretion of pro-inflammatory cytokines and decreased anti-inflammatory mediators. Salivary TNF-α, IL-6, and IL-10 may be potential non-invasive biomarkers that distinguish stroke patients from controls with high sensitivity and specificity. Of particular note is salivary TNF-α, which may indicate cognitive/physical impairment in post-stroke individuals. Further studies are needed in a larger stroke population to assess cytokines at both the salivary and central (plasma/serum) levels. Evaluation of other inflammatory mediators for their potential diagnostic significance also requires research. Declarations Author Contributions: Study concept and design: M.M. and K.G. Preparation of dental examination: P.G., K.G., and A.Z. Carrying out dental examination: P.G., K.G., and A.Z. Carrying out general examination: P.G and K.H. Collection of literature data: M.M and K.G. Analysis and interpretation of epidemiological data: M.M., K.G, and A.Z. Sample preparation: M.M and K.M.M. Laboratory determinations: M.M and K.M.M. Statistical analysis: M.M. Drafting of the manuscript: M.M, K.M.M. and K.G. Critical revision of the manuscript for important intellectual content: M.M., P.G., K.G., A.Z., and K.H. Study supervision: M.M., K.G. and A.Z. All authors approved the manuscript. All authors have read and agreed to the published version of the manuscript. Funding: This work was granted by the Medical University of Bialystok, Poland (grant number: SUB/1/DN/21/002/3330, SUB/1/DN/21/002/1209. Dr. Mateusz Maciejczyk was supported by the Foundation for Polish Science (FNP). Institutional Review Board Statement: The study was conducted according to the guidelines of the Declaration of Helsinki, and approved by the Ethics Committee of the Poznan University of Medical Sciences (resolutions 59/19 and 890/19) in Poland. Informed Consent Statement: Informed consent was obtained from all subjects involved in the study. Data Availability Statement: The datasets generated for this study are available on request to the corresponding author. Acknowledgments: The authors would like to thank the head and staff of the health care center (Bonifraterskie Centrum Zdrowia) in Piaski–Marysin (Piaski, Poland) for consent and help in organization of examination of the patients and all patients who participated in the research for their consent and cooperation. In addition, we would like to express our gratitude to neuropsychologist, Mrs. Hanna Norman, for her help in cognitive assessment of the patients. Conflicts of Interest: The authors declare no conflict of interest. References Guzik, A. & Bushnell, C. Stroke Epidemiology and Risk Factor Management. CONTINUUM Lifelong Learning in Neurology (2017) doi:10.1212/CON.0000000000000416. Jayaraj, R. L., Azimullah, S., Beiram, R., Jalal, F. Y. & Rosenberg, G. A. Neuroinflammation: friend and foe for ischemic stroke. J. Neuroinflammation 16 , 142 (2019). Feigin, V. L., Norrving, B. & Mensah, G. A. Global Burden of Stroke. Circ. Res. 120 , 439–448 (2017). 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Supplemental materials Supplementary material file is not available with this version. Additional Declarations No competing interests reported. Cite Share Download PDF Status: Under Review Version 1 posted Editorial decision: Major revision 03 Aug, 2021 Reviews received at journal 21 Jul, 2021 Reviewers agreed at journal 19 Jul, 2021 Reviewers agreed at journal 15 Jul, 2021 Reviewers invited by journal 16 Jun, 2021 Editor assigned by journal 16 Jun, 2021 Editor invited by journal 07 May, 2021 Submission checks completed at journal 06 May, 2021 First submitted to journal 05 May, 2021 You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. 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Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-494834","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":25292502,"identity":"79ecb9ee-e04f-4886-9982-8425820cb4d3","order_by":0,"name":"Mateusz Maciejczyk","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAABAUlEQVRIie2RsWrDMBCGTwl0EnhNoK9QMAQ0GflBstgI0iV+Aw8qhWbr3KGv0LXzgUCTqFZBlhqtHQyF0qn0nBZKBzsZC9U33C3/x89xAInEH4YDIC0saLArnA7PfymbQdEnKd+g+VpTwsXO4yy08jxjJr5y5+XDzlBLW6zHFOEUzLdW8aW2+YqHvXp0NSl20+gxBQflDHmODhTv90ogKUybccVHUj6Ql6QY3j8p4bsjSqCW5oZawLFrHlCKcKwlxNw0t4ov0K7YvVOVCNRSTd3i6y5u32SZ3ZnYv1hZCn/ZPfdtMaoM4GEuEGb00PqQrCbiP2Qa2DtAeVI4kUgk/hWfKC5pttDp+jgAAAAASUVORK5CYII=","orcid":"","institution":"Medical University of Białystok","correspondingAuthor":true,"prefix":"","firstName":"Mateusz","middleName":"","lastName":"Maciejczyk","suffix":""},{"id":25292503,"identity":"593e4f92-8fe3-4441-95a7-c74001f6d852","order_by":1,"name":"Kacper Mil","email":"","orcid":"","institution":"Medical University of Białystok","correspondingAuthor":false,"prefix":"","firstName":"Kacper","middleName":"","lastName":"Mil","suffix":""},{"id":25292504,"identity":"98d4cdc4-9733-4b3c-9294-5a4bec392872","order_by":2,"name":"Piotr Gerreth","email":"","orcid":"","institution":"Poznan University of Medical Sciences","correspondingAuthor":false,"prefix":"","firstName":"Piotr","middleName":"","lastName":"Gerreth","suffix":""},{"id":25292505,"identity":"0f2b811e-dd07-483b-ad98-32e8deffc589","order_by":3,"name":"Katarzyna Hojan","email":"","orcid":"","institution":"Poznan University of Medical Sciences","correspondingAuthor":false,"prefix":"","firstName":"Katarzyna","middleName":"","lastName":"Hojan","suffix":""},{"id":25292506,"identity":"7e1be749-92ab-41af-bb26-7f106e441f51","order_by":4,"name":"Anna Zalewska","email":"","orcid":"","institution":"Medical University of Białystok","correspondingAuthor":false,"prefix":"","firstName":"Anna","middleName":"","lastName":"Zalewska","suffix":""},{"id":25292507,"identity":"399ca34a-c615-4a27-b9ed-ae377f1da9bf","order_by":5,"name":"Karolina Gerreth","email":"","orcid":"","institution":"Poznan University of Medical Sciences","correspondingAuthor":false,"prefix":"","firstName":"Karolina","middleName":"","lastName":"Gerreth","suffix":""}],"badges":[],"createdAt":"2021-05-05 08:29:12","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-494834/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-494834/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":8930318,"identity":"7887d745-0eda-4bf6-a19e-bf96ab10c721","added_by":"auto","created_at":"2021-05-07 19:09:52","extension":"jpg","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":249524,"visible":true,"origin":"","legend":"Diagnostic significance of salivary TNF-α in ischemic stroke patients: correlation between salivary TNF-α and cognitive function in ACE III scale (A), results of ROC analysis for salivary TNF-α in relation to cognitive function status in ACE III scale (B-E). Differences statistically significant at: ** p \u003c 0.005**** p \u003c 0.0001","description":"","filename":"Figure1.jpg","url":"https://assets-eu.researchsquare.com/files/rs-494834/v1/2daacf69f3015ef432fad4c2.jpg"},{"id":13691694,"identity":"47002435-d991-45e5-ac20-cb08c9fbf5a8","added_by":"auto","created_at":"2021-09-17 12:39:46","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":559705,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-494834/v1/002fa4ce-d7d4-49a3-97b8-419eb47f2152.pdf"}],"financialInterests":"No competing interests reported.","formattedTitle":"Salivary Cytokine Profile in Patients with Ischemic Stroke. Can Salivary TNF-α be a Non-invasive Biomarker of Cognitive Impairment in Stroke Cases?","fulltext":[{"header":"Introduction","content":" \u003cp\u003eStroke is a clinical syndrome characterized by the sudden onset of focal and sometimes generalized brain damage with symptoms lasting longer than 24 hours \u003csup\u003e\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e\u003c/sup\u003e. According to the World Health Organization (WHO), stroke is the third leading cause of death in the adult population \u003csup\u003e\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e\u003c/sup\u003e. It is estimated that 5.5\u0026nbsp;million people die annually from this disease, accounting for 10% of all deaths \u003csup\u003e\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e\u003c/sup\u003e. Stroke is also the leading cause of long-term functional impairment and disability in adults \u003csup\u003e\u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e4\u003c/span\u003e\u003c/sup\u003e. There are two types of stroke: 10\u0026ndash;15% are hemorrhagic strokes, and the rest are ischemic strokes \u003csup\u003e\u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e\u003c/sup\u003e.\u003c/p\u003e \u003cp\u003eInflammation plays a crucial role in stroke pathogenesis \u003csup\u003e\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e,\u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e,\u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e7\u003c/span\u003e\u003c/sup\u003e. Acute cerebral ischemia induces cellular-molecular mechanisms involving both the central nervous system (CNS) and the vascular system. In response to ischemic brain injury, acute inflammatory reactions are induced, mediated by various cytokines, chemokines, and growth factors \u003csup\u003e\u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e8\u003c/span\u003e\u003c/sup\u003e. Among them, a unique role is attributed to TNF-α (tumor necrosis factor α), which is the first mediator synthesized in response to acute cerebral ischemia \u003csup\u003e\u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e\u003c/sup\u003e. Under these conditions, other pro-inflammatory cytokines are activated (e.g., interleukin-6 (IL-6) and interleukin-8 (IL-8)). TNF-α also upregulates the expression of matrix metalloproteinases (MMPs), which increases endothelial cell permeability and results in blood-brain barrier (BBB) dysfunction \u003csup\u003e\u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e9\u003c/span\u003e,\u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e10\u003c/span\u003e\u003c/sup\u003e. Pro-inflammatory mediators exert direct damaging effects on neurons and astrocytes and promote prostaglandin release and leukocyte migration \u003csup\u003e\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e,\u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e,\u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e7\u003c/span\u003e\u003c/sup\u003e. It has been shown that IL-6 and TNF-α stimulate the synthesis and secretion of several chemotactic factors, which activate and attract peripheral blood leukocytes (neutrophils and monocytes) to the site of stroke lesions \u003csup\u003e\u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e9\u003c/span\u003e,\u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e11\u003c/span\u003e\u003c/sup\u003e. Therefore, adhesion molecules' expression on the endothelial surface is increased, which potentiates leukocyte penetration into the ischemic brain area. Simultaneously, the secretion of interleukin-10 (IL-10) increases, inhibiting the inflammatory response \u003csup\u003e\u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e12\u003c/span\u003e\u003c/sup\u003e.\u003c/p\u003e \u003cp\u003eStroke diagnosis is based on physical examination and imaging studies (CT) \u003csup\u003e\u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e13\u003c/span\u003e\u003c/sup\u003e. Despite the precise definition of stroke, up to 20\u0026ndash;25% of diagnoses may be conditions that mimic stroke \u003csup\u003e\u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e14\u003c/span\u003e\u003c/sup\u003e. Availability of the latest diagnostic techniques (CT perfusion, MRI) is also often limited \u003csup\u003e\u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e15\u003c/span\u003e\u003c/sup\u003e. Therefore, it is not surprising that new diagnostic methods are still being sought \u003csup\u003e\u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e16\u003c/span\u003e\u003c/sup\u003e. In particular, biochemical biomarkers are being searched extensively to facilitate early diagnosis of stroke or to establish further patient\u0026rsquo;s prognosis \u003csup\u003e\u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e17\u003c/span\u003e,\u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e18\u003c/span\u003e\u003c/sup\u003e. Given the critical role of inflammation in stroke pathogenesis, the use of cytokines and chemokines in laboratory diagnostics has been increasingly advocated \u003csup\u003e\u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e16\u003c/span\u003e,\u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e19\u003c/span\u003e\u003c/sup\u003e. Of particular note is the evaluation of TNF-α and IL-6, whose serum levels increase in the acute phase of stroke as well as correlate with its cerebrospinal fluid (CSF) levels and stroke focus size \u003csup\u003e\u003cspan additionalcitationids=\"CR21 CR22\" citationid=\"CR20\" class=\"CitationRef\"\u003e20\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e23\u003c/span\u003e\u003c/sup\u003e. The high diagnostic utility also seems to be demonstrated by IL-10, whose levels decrease in patients with ischemic stroke and indicate the predominance of pro-inflammatory processes in the early stroke phase \u003csup\u003e\u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e12\u003c/span\u003e,\u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e24\u003c/span\u003e\u003c/sup\u003e.\u003c/p\u003e \u003cp\u003eNowadays, the use of biological materials collected in a non-invasive manner, without special equipment and medical personnel, is gaining interest in laboratory diagnostics \u003csup\u003e\u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e25\u003c/span\u003e,\u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e26\u003c/span\u003e\u003c/sup\u003e. This reduces the patient's anxiety associated with the examination, promotes more frequent monitoring of health condition, and thus diagnoses the disorder at an early stage \u003csup\u003e\u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e27\u003c/span\u003e\u003c/sup\u003e. Saliva is an interesting alternative to blood or CSF that are commonly used in laboratory medicine \u003csup\u003e\u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e25\u003c/span\u003e,\u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e28\u003c/span\u003e\u003c/sup\u003e. Saliva is a secretion of major and minor salivary glands containing water (94\u0026ndash;99%) and numerous proteins, lipids, carbohydrates, hormones, immunoglobulins, cytokines, growth factors, and vitamins \u003csup\u003e\u003cspan additionalcitationids=\"CR30\" citationid=\"CR29\" class=\"CitationRef\"\u003e29\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e31\u003c/span\u003e\u003c/sup\u003e. Its composition is therefore very similar to blood plasma. Indeed, the level of many compounds in saliva correlates with their content in blood/CSF, indicating a solid diagnostic potential \u003csup\u003e25,28,32\u0026minus;34\u003c/sup\u003e. Although salivary cytokines are used to diagnose many systemic diseases \u003csup\u003e\u003cspan additionalcitationids=\"CR36 CR37 CR38\" citationid=\"CR35\" class=\"CitationRef\"\u003e35\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR39\" class=\"CitationRef\"\u003e39\u003c/span\u003e\u003c/sup\u003e, there is no data on salivary inflammatory mediators' diagnostic utility in stroke individuals. Considering the continuous increase in stroke morbidity/mortality and the lack of non-invasive biomarkers of the disease, our study aimed to evaluate TNF-α, IL-6, and IL-10 levels in the non-stimulated (NWS) and stimulated (SWS) whole saliva of stroke patients.\u003c/p\u003e "},{"header":"Material And Methods","content":"\u003cdiv id=\"Sec3\" class=\"Section2\"\u003e\n\u003ch2\u003eStudy population\u003c/h2\u003e\n\u003cp\u003eThe study was carried out from June to August 2019 in a health center (Bonifraterskie Centrum Zdrowia) in Piaski\u0026ndash;Marysin (Piaski, Poland). In this center, patients with various disorders are hospitalized, including the individuals after the cerebral stroke that arrive from different country provinces. At the time of the research, there were 315 patients in the neurorehabilitation unit following different incidents, including vascular brain damage, brain injury, spinal cord injury, surgically treated patients with a brain tumor, sclerosis multiplex, myelopathy, and polyneuropathy. It was determined that 218 (65.71%) patients were stroke survivors, and the individuals were admitted to the neurorehabilitation ward immediately from the hospital, directly after the acute phase cessation, in a subacute phase of the stroke. A medical doctor assessed each patient at the beginning of the stay at the unit, and afterward, he/she was suggested to comprehensive individual and similar rehabilitation. Most of the patients were able to cooperate, understand instructions, and communicate. In addition, the majority of the individuals followed the same diet, including a baseline diet for most patients or a diet for people with diabetes mellitus. All the meals were prepared in the hospital and were distributed to the patients at the same time daily.\u003c/p\u003e\n\u003cp\u003eDecisively, 97 (30.79%) individuals with other diseases than stroke were excluded from the study, and 25 (11.47% of stroke survivors; 7.94% of all individuals rehabilitated at the health center fully completed the examination, saliva sampling, and were considered in the analysis (Table\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e1\u003c/span\u003e). The other stroke patients had to be excluded from the study because of the following reasons: 80 (36.70%) patients refused to participate in the study; 42 (19.27%) people were uncooperative since they could not give conscious written informed consent for participation in the research and/or entirely communicate; 28 (12.84%) patients did not report for a sampling of saliva and dental examination, although they gave informed consent and were reminded up to 4 times; 11 (5.04%) subjects withdrew from the research after sampling of non-stimulated saliva due to physiological or psychological fatigue; 5 (2.29%) people were not able to gather the saliva due to issues concerning understanding the procedure because of cognitive and language deficits, and 2 (0.92%) patients were transferred to another hospital because of deterioration of general health.\u003c/p\u003e\n\u003cp\u003eEssential information on the general condition and health status of patients were obtained from individuals' files and contained: gender, age, the medication used, time since diagnosis of cerebral stroke, and medical history (Table\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e1\u003c/span\u003e).\u003c/p\u003e\n\u003cdiv class=\"gridtable\"\u003e\n\u003ctable id=\"Tab1\" border=\"1\"\u003e\u003ccaption\u003e\n\u003cdiv class=\"CaptionNumber\"\u003eTable 1\u003c/div\u003e\n\u003cdiv class=\"CaptionContent\"\u003e\n\u003cp\u003eGeneral characteristics of patients.\u003c/p\u003e\n\u003c/div\u003e\n\u003c/caption\u003e\n\u003cthead\u003e\n\u003ctr\u003e\n\u003cth rowspan=\"2\" align=\"left\"\u003e\u0026nbsp;\u003c/th\u003e\n\u003cth colspan=\"2\" align=\"left\"\u003e\n\u003cp\u003eControl\u003c/p\u003e\n\u003cp\u003e\u003cem\u003en\u003c/em\u003e\u0026thinsp;=\u0026thinsp;25\u003c/p\u003e\n\u003c/th\u003e\n\u003cth colspan=\"2\" align=\"left\"\u003e\n\u003cp\u003eStroke\u003c/p\u003e\n\u003cp\u003e\u003cem\u003en\u003c/em\u003e\u0026thinsp;=\u0026thinsp;25\u003c/p\u003e\n\u003c/th\u003e\n\u003cth rowspan=\"2\" align=\"left\"\u003e\n\u003cp\u003ep value\u003c/p\u003e\n\u003c/th\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003eMean\u0026thinsp;\u0026plusmn;\u0026thinsp;SD\u003c/p\u003e\n\u003c/th\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003e95%CI\u003c/p\u003e\n\u003c/th\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003eMean\u0026thinsp;\u0026plusmn;\u0026thinsp;SD\u003c/p\u003e\n\u003c/th\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003e95%CI\u003c/p\u003e\n\u003c/th\u003e\n\u003c/tr\u003e\n\u003c/thead\u003e\n\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eSex (male/female)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd colspan=\"2\" align=\"left\"\u003e\n\u003cp\u003e12/13\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd colspan=\"2\" align=\"left\"\u003e\n\u003cp\u003e12/13\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e\u0026gt;\u0026thinsp;0.9999\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eAge\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e63.52\u0026thinsp;\u0026plusmn;\u0026thinsp;8.545\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e59.99\u0026ndash;67.05\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e63.52\u0026thinsp;\u0026plusmn;\u0026thinsp;8.545\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e59.99\u0026ndash;67.05\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e\u0026gt;\u0026thinsp;0.9999\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd colspan=\"6\" align=\"left\"\u003e\n\u003cp\u003e\u003cem\u003eCognitive and physical functional status\u003c/em\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eACE III\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e97.24\u0026thinsp;\u0026plusmn;\u0026thinsp;1.422\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e96.65\u0026ndash;97.83\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e69.8\u0026thinsp;\u0026plusmn;\u0026thinsp;22.97\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e60.32\u0026ndash;79.28\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e\u0026lt;\u0026thinsp;0.0001\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eBI\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e20\u0026thinsp;\u0026plusmn;\u0026thinsp;0\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e20\u0026ndash;20\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e10.92\u0026thinsp;\u0026plusmn;\u0026thinsp;3.662\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e9.408\u0026ndash;12.43\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e\u0026lt;\u0026thinsp;0.0001\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eBBS\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e55.52\u0026thinsp;\u0026plusmn;\u0026thinsp;0.5099\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e55.31\u0026ndash;55.73\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e31\u0026thinsp;\u0026plusmn;\u0026thinsp;16.27\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e24.28\u0026ndash;37.72\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e\u0026lt;\u0026thinsp;0.0001\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eFIM\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e125.3\u0026thinsp;\u0026plusmn;\u0026thinsp;0.6904\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e125\u0026ndash;125.6\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e86.88\u0026thinsp;\u0026plusmn;\u0026thinsp;30.31\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e74.37\u0026ndash;99.39\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e\u0026lt;\u0026thinsp;0.0001\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd colspan=\"6\" align=\"left\"\u003e\n\u003cp\u003e\u003cem\u003eComorbidities\u003c/em\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003ehypertension n (%)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd colspan=\"2\" align=\"left\"\u003e\n\u003cp\u003e18 (72)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd colspan=\"2\" align=\"left\"\u003e\n\u003cp\u003e17 (68)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e\u0026gt;\u0026thinsp;0.9999\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003ediabetes n (%)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd colspan=\"2\" align=\"left\"\u003e\n\u003cp\u003e4 (16)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd colspan=\"2\" align=\"left\"\u003e\n\u003cp\u003e4 (16)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e\u0026gt;\u0026thinsp;0.9999\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eepilepsy n (%)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd colspan=\"2\" align=\"left\"\u003e\n\u003cp\u003e4 (16)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd colspan=\"2\" align=\"left\"\u003e\n\u003cp\u003e3 (12)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e\u0026gt;\u0026thinsp;0.9999\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003earteriosclerosis n (%)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd colspan=\"2\" align=\"left\"\u003e\n\u003cp\u003e7 (28)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd colspan=\"2\" align=\"left\"\u003e\n\u003cp\u003e8 (32)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e\u0026gt;\u0026thinsp;0.9999\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003elimb thrombosis n (%)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd colspan=\"2\" align=\"left\"\u003e\n\u003cp\u003e2 (8)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd colspan=\"2\" align=\"left\"\u003e\n\u003cp\u003e2 (8)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e\u0026gt;\u0026thinsp;0.9999\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eatrial fibrillation n (%)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd colspan=\"2\" align=\"left\"\u003e\n\u003cp\u003e3 (12)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd colspan=\"2\" align=\"left\"\u003e\n\u003cp\u003e3 (12)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e\u0026gt;\u0026thinsp;0.9999\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd colspan=\"6\" align=\"left\"\u003e\n\u003cp\u003e\u003cem\u003eDrugs\u003c/em\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e\u0026lt;\u0026thinsp;5 drugs/day \u003cem\u003en\u003c/em\u003e (%)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd colspan=\"2\" align=\"left\"\u003e\n\u003cp\u003e10 (40)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd colspan=\"2\" align=\"left\"\u003e\n\u003cp\u003e9 (36)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e\u0026gt;\u0026thinsp;0.9999\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e\u0026ge;\u0026thinsp;5 drugs/day \u003cem\u003en\u003c/em\u003e (%)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd colspan=\"2\" align=\"left\"\u003e\n\u003cp\u003e15 (60)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd colspan=\"2\" align=\"left\"\u003e\n\u003cp\u003e16 (64)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e\u0026gt;\u0026thinsp;0.9999\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003c/tbody\u003e\n\u003c/table\u003e\n\u003c/div\u003e\n\u003cp\u003eAbbreviations:\u0026nbsp;ACE III \u0026ndash; Addenbrooke\u0026rsquo;s Cognitive Examination; BI \u0026ndash; Barthel Index; BBS \u0026ndash; Berg Balance Scale; FIM \u0026ndash; functional independence measure; differences statistically significant at: * p\u0026thinsp;\u0026lt;\u0026thinsp;0.05; ** p\u0026thinsp;\u0026lt;\u0026thinsp;0.005; *** p\u0026thinsp;\u0026lt;\u0026thinsp;0.0005; **** p\u0026thinsp;\u0026lt;\u0026thinsp;0.0001.\u003c/p\u003e\n\u003cp\u003eFor measurement of the functional status of the individuals the following scales were used:\u003c/p\u003e\n\u003col\u003e\n\u003cli\u003e\n\u003cp\u003eAddenbrooke\u0026rsquo;s Cognitive Examination III (ACE III) for differentiation between patients without and with cognitive impairment \u003csup\u003e\u003cspan class=\"CitationRef\"\u003e40\u003c/span\u003e\u003c/sup\u003e.\u003c/p\u003e\n\u003c/li\u003e\n\u003cli\u003e\n\u003cp\u003eThe Barthel Index (BI) to measure performance in activities of daily living (ADL)\u003csup\u003e\u003cspan class=\"CitationRef\"\u003e41\u003c/span\u003e\u003c/sup\u003e.\u003c/p\u003e\n\u003c/li\u003e\n\u003cli\u003e\n\u003cp\u003eThe Berg Balance Scale (BBS) to determine an individual's ability or inability to safely balance during a series of predetermined tasks \u003csup\u003e\u003cspan class=\"CitationRef\"\u003e42\u003c/span\u003e\u003c/sup\u003e.\u003c/p\u003e\n\u003c/li\u003e\n\u003cli\u003e\n\u003cp\u003eThe functional independence measure (FIM) for exploration of the patient's psychological, physical, and social functioning \u003csup\u003e\u003cspan class=\"CitationRef\"\u003e43\u003c/span\u003e\u003c/sup\u003e.\u003c/p\u003e\n\u003c/li\u003e\n\u003c/ol\u003e\n\u003cp\u003eOne experienced medical doctor, a neurorehabilitation specialist, qualified all the patients for the examination according to study criteria (Table\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e2\u003c/span\u003e).\u003c/p\u003e\n\u003cdiv class=\"gridtable\"\u003e\n\u003ctable id=\"Tab2\" border=\"1\"\u003e\u003ccaption\u003e\n\u003cdiv class=\"CaptionNumber\"\u003eTable 2\u003c/div\u003e\n\u003cdiv class=\"CaptionContent\"\u003e\n\u003cp\u003eInclusion and exclusion criteria for the subjects in the study group.\u003c/p\u003e\n\u003c/div\u003e\n\u003c/caption\u003e\n\u003cthead\u003e\n\u003ctr\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003eInclusion Criteria\u003c/p\u003e\n\u003c/th\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003eExclusion Criteria\u003c/p\u003e\n\u003c/th\u003e\n\u003c/tr\u003e\n\u003c/thead\u003e\n\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eage of consent (\u0026gt;\u0026thinsp;18 years)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003enonage (\u0026lt;\u0026thinsp;18 years)\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003econsciousness\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eunconsciousness\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003egiving of written and informed consent for saliva sampling and oral examination\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003elack of written and informed consent for saliva sampling and oral examination from patient\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003egood general condition\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003epoor general condition\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003econfirmed cerebral infarction or cerebral hemorrhage based on CT and magnetic resonance imaging (MRI)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eunconfirmed cerebral infarction or cerebral hemorrhage based on CT and magnetic resonance imaging (MRI)\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eadequate capacity to follow instructions, i.e., understanding how to perform the procedures, ability to collect a saliva sample, and being able to answer questions during the examination\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003einadequate capacity to follow instructions, i.e., not understanding how to perform the procedures, inability to collect a saliva sample, and being unable to answer questions during the examination\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003erecovery from the acute phase of ischemic or hemorrhagic stroke in all brain areas\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eno recovery from the acute phase of ischemic or hemorrhagic stroke in all brain areas\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eno stroke recurrence during the subacute phase\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003estroke recurrence during the subacute phase\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003egood cooperation\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003einsufficient cooperation due to cognitive and/or language deficits\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eischemic stroke treated without thrombectomy or thrombolysis\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eischemic stroke treated with thrombectomy or thrombolysis\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003ecapability to collect a saliva sample\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eincapability to collect a saliva sample\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eno legal guardianship\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003elegal guardianship\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003epatients without psychiatric or cognitive disorders\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003epatients with psychiatric or cognitive disorders\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eno autoimmune disease (rheumatoid arthritis, systemic lupus erythematosus)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eautoimmune disease (rheumatoid arthritis, systemic lupus erythematosus)\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eno lung disease (chronic obstructive pulmonary disease) or cardiovascular disease (angina or uncontrolled hypertension)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003elung disease (chronic obstructive pulmonary disease) or cardiovascular disease (angina or uncontrolled hypertension)\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003egood general condition\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eheart failure above NYHA II\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eno intake of dietary supplements and vitamins for the last three months\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eintake of dietary supplements and vitamins for the last three months\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003enon-smokers\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003esmokers\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003ewomen/men without malnutrition\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003epatients with malnutrition, i.e., having body mass index\u0026thinsp;\u0026lt;\u0026thinsp;18kg/m\u003csup\u003e2\u003c/sup\u003e or with weight loss\u0026thinsp;\u0026gt;\u0026thinsp;10% during the previous three months\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003ethe first admission to cure stroke unit was more than 5\u0026ndash;6 (to 10) hours from the onset of the early neurological symptoms\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n\u003c/tr\u003e\n\u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec4\" class=\"Section3\"\u003e\n\u003ch2\u003eControl group\u003c/h2\u003e\n\u003cp\u003eThe control group included 25 generally healthy patients who reported for oral examination to the Department of Restorative Dentistry of the Medical University of Bialystok (Bialystok, Poland) from March to September 2017. Controls were similar to the stroke patients regarding age and gender, oral hygiene, dentition, and periodontium status (Table\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e1\u003c/span\u003e). All participants were provided with medical clearance by clinicians before involvement in the research. Individuals from the control group followed a regular, balanced diet (which was not restricted), and they were given standard physical activity recommendations.\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec5\" class=\"Section3\"\u003e\n\u003ch2\u003eSampling of saliva\u003c/h2\u003e\n\u003cp\u003eThe research material was total mixed non-stimulated saliva (NWS) and stimulated saliva (SWS), and both samples were collected via spitting. Saliva sampling was done in the health center during summertime, i.e., between June and September, to keep similar weather conditions outside. Before the procedure, individuals were instructed not to intake any solid food and/or liquid, other than clean water, at least two hours before saliva sampling. All individuals were in the subacute phase of the stroke. The study and control group individuals had not taken any medication eight hours before sampling of saliva \u003csup\u003e\u003cspan class=\"CitationRef\"\u003e44\u003c/span\u003e,\u003cspan class=\"CitationRef\"\u003e45\u003c/span\u003e\u003c/sup\u003e. The oral bioliquid was gathered, from subjects that had restrained from intensive physical activity for the preceding twelve hours, i.e., between 7:30 a.m. and 9:00 a.m. The patients were also indicated not to carry out any oral hygiene practices (i.e., mouth rinsing, teeth brushing, gum chewing, etc.) \u003csup\u003e\u003cspan class=\"CitationRef\"\u003e45\u003c/span\u003e,\u003cspan class=\"CitationRef\"\u003e46\u003c/span\u003e\u003c/sup\u003e. The saliva was collected after a five‐minute adaptation to the environment in a separate, private room. The oral cavity was rinsed two times with distilled water at room temperature before gathering saliva to avoid possible impurity from other sources. During the procedure, subjects were seated in an adjustable chair, individually adapted to each individual's height, with the head slightly bent downwards and resting in a convenient position, and they tried to limit their lips and face movements \u003csup\u003e\u003cspan class=\"CitationRef\"\u003e45\u003c/span\u003e,\u003cspan class=\"CitationRef\"\u003e46\u003c/span\u003e\u003c/sup\u003e. The samples of saliva that were stored during the first minute were ejected. Saliva was accumulated into a sterile Falcon tube placed in a container with ice. The non‐stimulated saliva was collected for 10 minutes and stimulated saliva was gathered in the same manner for 5 minutes to avoid the patients' physiological and/or psychological fatigue. The secretion of this oral bioliquid was stimulated by ten \u0026micro;L of 2% citric acid applied to the tongue's central part every 30 seconds. Afterward, the volume of saliva was measured using a calibrated pipette with an accuracy of 0.1 mL \u003csup\u003e\u003cspan class=\"CitationRef\"\u003e47\u003c/span\u003e\u003c/sup\u003e. The minute flow of NWS and SWS was measured by dividing the saliva volume by the time essential for its secretion (mL/min). Forthwith, after collection of saliva, it was centrifuged (+\u0026thinsp;4\u0026deg;C, 20 min, 3000 \u0026times; g; MPW 351, MPW Med. Instruments, Warsaw, Poland). To protect againts sample oxidation, butylated hydroxytoluene (BHT, Sigma‐Aldrich, Saint Louis, MO, USA) was added into the acquired supernatants in the amount of 10 \u0026micro;L 0.5 M BHT in acetonitrile (ACN)/1 mL of saliva \u003csup\u003e\u003cspan class=\"CitationRef\"\u003e48\u003c/span\u003e\u003c/sup\u003e. The samples of NWS and SWS were frozen at \u0026minus;\u0026thinsp;80\u0026deg;C and stored for no more than three months for further investigation.\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec6\" class=\"Section3\"\u003e\n\u003ch2\u003eOral examination\u003c/h2\u003e\n\u003cp\u003eThe dentition was evaluated in a separate room shortly after saliva sampling. According to the World Health Organization criteria, the individual's check-up was carried out using a plain mouth mirror and a dental probe in artificial lighting \u003csup\u003e\u003cspan class=\"CitationRef\"\u003e49\u003c/span\u003e,\u003cspan class=\"CitationRef\"\u003e50\u003c/span\u003e\u003c/sup\u003e. The dentist performed the dental examination in front of the chair where the patient was seated and with the individual's head resting against the wall. Each accessible tooth surface was assessed and scored as generally healthy, decayed (DT), extracted due to the carious process (MT), or filled because of caries (FT). The data collected were used to calculate the DMFT index, that expresses dental caries experience, and it is the sum of the following components: DT, MT, and FT. Additionally, the prevalence of dental caries was determined as a percentage of patients with DMFT\u0026thinsp;\u0026gt;\u0026thinsp;0. Gingival index (GI) and Plaque Index (PlI) were evaluated on the teeth 16, 12, 24, 36, 32, 44 using four-degree scales (from 0 to 3) \u003csup\u003e\u003cspan class=\"CitationRef\"\u003e51\u003c/span\u003e\u003c/sup\u003e. The oral examination was performed by two dentists (P.G. and K.G.), subsequently after sampling of non-stimulated (NWS) and stimulated (SWS) whole saliva, and after previous training and calibration by an experienced dental specialist (A.Z.). The inter‐examiner and intra‐examiner agreement for DMFT, PlI and GI was evaluated by another dental examination in 10 patients with a \u0026kappa;\u0026thinsp;\u0026gt;\u0026thinsp;0.94.\u003c/p\u003e\n\u003c/div\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec7\" class=\"Section2\"\u003e\n\u003ch2\u003ePatient\u0026rsquo;s consent and Bioethical Issues\u003c/h2\u003e\n\u003cp\u003eAll individuals, i.e., stroke patients and healthy controls, voluntarily took part in the study, and they were provided with comprehensive information concerning its purpose, procedures, risks, and benefits. Full written consent was obtained from all participants in accordance with the Declaration of Helsinki for a sampling of saliva and dental examination.\u003c/p\u003e\n\u003cp\u003eBefore the research, it was approved by the Bioethics Committee of the Poznan University of Medical Sciences (resolutions 59/19 and 890/19).\u003c/p\u003e\n\u003cdiv id=\"Sec8\" class=\"Section3\"\u003e\n\u003ch2\u003eSalivary cytokine profile\u003c/h2\u003e\n\u003cp\u003eSalivary concentrations of tumor necrosis factor \u0026alpha; (TNF-\u0026alpha;), interleukin 6 (IL-6), and interleukin 10 (IL-10) were assessed using commercial enzyme-linked immunosorbent assay (ELISA) kits according to the manufacturer\u0026rsquo;s instructions (EIAab, Wuhan, China). Briefly, specific antibodies suitably labeled with an enzyme were added to the antigen-coated plate. Depending on the amount of antigen, antibodies specifically bound to the antigen and unbound antibodies were eluted. After adding the substrate/chromophore, the enzyme-catalyzed the reaction, and the colored reaction product was measured colorimetrically at 405 nm using Mindray MR-96 Microplate Reader. All determinations were performed in duplicate samples.\u003c/p\u003e\n\u003cp\u003eSalivary total protein content was determined in duplicate samples by the bicinchoninic acid (BCA) method (Pierce BCA Protein Assay Kit; Rockford, IL, USA). All results were presented as concentration (pg/mL), salivary output (pg/min), and specific content (mg/mg protein). The TNF-\u0026alpha; /IL-10 and IL-6/IL-10 ratios were also calculated.\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec9\" class=\"Section3\"\u003e\n\u003ch2\u003eStatistical data analysis\u003c/h2\u003e\n\u003cp\u003eStatistical analysis was carried out using GraphPad Prism 8 for macOS (GraphPad Software, La Jolla, CA, USA). Due to the fact that most of the results were characterized by a normal distribution (Kolmogorov-Smirnov test), the t-student test was used to compare two groups. If the distribution was not normal, the Mann-Whitney U test was used. ANOVA analysis of variance with Tukey's post hoc test was used to compare the 3 groups, and p values were calculated with correction for multiple comparisons. Correlations between salivary biomarkers and clinical data were performed using the Pearson correlation coefficient. The results were presented as mean\u0026thinsp;\u0026plusmn;\u0026thinsp;SD and 95% confidence interval (95% CI). The assumed statistical significance was p\u0026thinsp;\u0026lt;\u0026thinsp;0.05.\u003c/p\u003e\n\u003cp\u003eCohen Kappa (online calculator) was used to establish the inter-and intracorporeal agreement between the evaluated dental indicators.\u003c/p\u003e\n\u003cp\u003eThe diagnostic usefulness of salivary cytokines was determined using ROC (Receiver Operating Curve) analysis by evaluating the area under the curve (AUC). The sensitivity and specificity and the cut-off point, which was characterized by the highest sensitivity and specificity, were given.\u003c/p\u003e\n\u003cp\u003eThe number of patients was set \u003cem\u003ea priori\u003c/em\u003e based on our previous study. For this purpose, an online sample size calculator (ClinCalc) was used. The level of significance was set at 0.05, and the power of the study was 0.9. Variables used for sample size calculation were concentration of salivary IL-6 and IL-10. The minimum number of patients was 21 (for one group).\u003c/p\u003e\n\u003c/div\u003e\n\u003c/div\u003e"},{"header":"Results","content":"\u003cdiv class=\"gridtable\"\u003e\n\u003ctable id=\"Tab3\" border=\"1\"\u003e\u003ccaption\u003e\n\u003cdiv class=\"CaptionNumber\"\u003eTable 3\u003c/div\u003e\n\u003cdiv class=\"CaptionContent\"\u003e\n\u003cp\u003eSalivary gland function and dental characteristics of the stroke and healthy controls.\u003c/p\u003e\n\u003c/div\u003e\n\u003c/caption\u003e\n\u003cthead\u003e\n\u003ctr\u003e\n\u003cth colspan=\"2\" rowspan=\"2\" align=\"left\"\u003e\u0026nbsp;\u003c/th\u003e\n\u003cth colspan=\"2\" align=\"left\"\u003e\n\u003cp\u003eControl\u003c/p\u003e\n\u003cp\u003e\u003cem\u003en\u003c/em\u003e\u0026thinsp;=\u0026thinsp;25\u003c/p\u003e\n\u003c/th\u003e\n\u003cth colspan=\"2\" align=\"left\"\u003e\n\u003cp\u003eStroke\u003c/p\u003e\n\u003cp\u003e\u003cem\u003en\u003c/em\u003e\u0026thinsp;=\u0026thinsp;25\u003c/p\u003e\n\u003c/th\u003e\n\u003cth rowspan=\"2\" align=\"left\"\u003e\n\u003cp\u003ep value\u003c/p\u003e\n\u003c/th\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003eMean\u0026thinsp;\u0026plusmn;\u0026thinsp;SD\u003c/p\u003e\n\u003c/th\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003e95%CI\u003c/p\u003e\n\u003c/th\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003eMean\u0026thinsp;\u0026plusmn;\u0026thinsp;SD\u003c/p\u003e\n\u003c/th\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003e95%CI\u003c/p\u003e\n\u003c/th\u003e\n\u003c/tr\u003e\n\u003c/thead\u003e\n\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd rowspan=\"2\" align=\"left\"\u003e\n\u003cp\u003eNWS\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eFlow rate (mL/min)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.33\u0026thinsp;\u0026plusmn;\u0026thinsp;0.08461\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.2951\u0026ndash;0.3649\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.394\u0026thinsp;\u0026plusmn;\u0026thinsp;0.2459\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.2925\u0026ndash;0.4955\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.2245\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eTotal protein (\u0026micro;g/mL)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e1220\u0026thinsp;\u0026plusmn;\u0026thinsp;174\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e1148\u0026ndash;1292\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e1141\u0026thinsp;\u0026plusmn;\u0026thinsp;281.5\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e1025\u0026ndash;1257\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.2391\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd rowspan=\"2\" align=\"left\"\u003e\n\u003cp\u003eSWS\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eFlow rate (mL/min)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.9112\u0026thinsp;\u0026plusmn;\u0026thinsp;0.2629\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.8027-1.02\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.5796\u0026thinsp;\u0026plusmn;\u0026thinsp;0.2454\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.4783\u0026ndash;0.6809\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e\u0026lt;\u0026thinsp;0.0001\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eTotal protein (\u0026micro;g/mL)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e1310\u0026thinsp;\u0026plusmn;\u0026thinsp;172.3\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e1239\u0026ndash;1382\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e947\u0026thinsp;\u0026plusmn;\u0026thinsp;244.8\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e846\u0026ndash;1048\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e\u0026lt;\u0026thinsp;0.0001\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd colspan=\"2\" align=\"left\"\u003e\n\u003cp\u003eDMFT\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e24.4\u0026thinsp;\u0026plusmn;\u0026thinsp;7.848\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e21.16\u0026ndash;27.64\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e23.08\u0026thinsp;\u0026plusmn;\u0026thinsp;7.989\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e19.78\u0026ndash;26.38\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.5584\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd colspan=\"2\" align=\"left\"\u003e\n\u003cp\u003eGI\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.8136\u0026thinsp;\u0026plusmn;\u0026thinsp;0.8085\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.4799\u0026ndash;1.147\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.72\u0026thinsp;\u0026plusmn;\u0026thinsp;0.7294\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.4189\u0026ndash;1.021\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.6693\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd colspan=\"2\" align=\"left\"\u003e\n\u003cp\u003ePlI\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e1.254\u0026thinsp;\u0026plusmn;\u0026thinsp;0.9743\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.8514\u0026ndash;1.656\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e1.234\u0026thinsp;\u0026plusmn;\u0026thinsp;0.9953\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.8228\u0026ndash;1.644\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.9431\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003eAbbreviations: NWS \u0026ndash; non-stimulated whole saliva; SWS \u0026ndash; stimulated whole saliva; DMFT index - a sum of decayed teeth (DT), teeth missing due to carious process (MT), and teeth filled because of caries (FT); GI \u0026ndash; Gingival Index; PlI \u0026ndash; Plaque Index.\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec11\" class=\"Section2\"\u003e\n\u003ch2\u003eSalivary gland function\u003c/h2\u003e\n\u003cp\u003eThe secretion of non-stimulated saliva did not differ significantly between the study and control groups. Stimulated saliva secretion was significantly decreased (\u0026darr;36%) in the stroke population than the controls (Table\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e3\u003c/span\u003e).\u003c/p\u003e\n\u003cp\u003eThe concentration of total protein in non-stimulated saliva did not differ significantly between healthy controls and stroke individuals. The total protein concentration in stimulated saliva of the stroke population was significantly lower (\u0026darr;28%) than in the control group (Table\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e3\u003c/span\u003e).\u003c/p\u003e\n\u003cdiv id=\"Sec12\" class=\"Section3\"\u003e\n\u003ch2\u003eOral health status\u003c/h2\u003e\n\u003cp\u003eDMFT, GI, and PlI were not significantly different between the study and control group (Table\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e3\u003c/span\u003e).\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec13\" class=\"Section3\"\u003e\n\u003ch2\u003eSalivary cytokines\u003c/h2\u003e\n\u003cp\u003eNon-stimulated saliva of stroke patients was characterized by significantly higher concentration (\u0026uarr;135%), output (\u0026uarr;196%), and specific amount (\u0026uarr;170%) of TNF-\u0026alpha;. Similarly, these parameters were increased regarding IL-6 (respectively: \u0026uarr;85%, \u0026uarr;124%, 103%). On the other hand, the concentration (\u0026darr;47%), output (\u0026darr;78%), and specific amount (\u0026darr;42%) of IL-10 decreased. As a result of these changes, the TNF-\u0026alpha;/IL-10 ratio (\u0026uarr;507%) and IL-6/IL-10 ratio (\u0026uarr;378%) increased markedly.\u003c/p\u003e\n\u003cp\u003eThe results of measurements in stimulated saliva were comparable to those in non-stimulated saliva. The concentration (\u0026uarr;68%) and specific amount (\u0026uarr;147%) of TNF-\u0026alpha; were significantly higher in patients with stroke than in control. Mentioned parameters were also higher in relation to IL-6 (concentration: \u0026uarr;62%, specific amount: \u0026uarr;57%). The output of TNF-\u0026alpha; and IL-6 remained statistically insignificant. IL-10 concentration, output, and specific amount decreased, similarly to NWS (respectively: \u0026darr;65%, \u0026uarr;36%, \u0026uarr;48%). In consequence, TNF-\u0026alpha;/IL-10 (\u0026uarr;611%) and IL-6/IL-10 (\u0026uarr;568%) ratios increased greatly (Table\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e4\u003c/span\u003e).\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec14\" class=\"Section3\"\u003e\n\u003ch2\u003eCorrelations\u003c/h2\u003e\n\u003cp\u003eStatistically significant correlations were summed up in Table\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e5\u003c/span\u003e. The positive correlations were revealed only in non-stimulated saliva between IL-6 concentration and ADL, and IL-6 specific amount and ADL.\u003c/p\u003e\n\u003cp\u003eThe strong negative correlations in non-stimulated saliva were observed between TNF-\u0026alpha; concentration and BBS, TNF-\u0026alpha; concentration and ACE III or TNF-\u0026alpha; concentration and ADL. Moreover, IL-6 specific amount and PlI and IL-6 output and FIM correlate negatively in unstimulated saliva. The negative correlation between TNF-\u0026alpha; output and BBS was noted in either stimulated or non-stimulated saliva.\u003c/p\u003e\n\u003cp\u003eIn stimulated saliva, TNF-\u0026alpha; output correlated negatively with BBS, ADL and ACE III. On the other hand, IL-6 output was observed to correlate negatively with BBS and ADL.\u003c/p\u003e\n\u003cp\u003eResults of all correlations were presented in the supplementary material.\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec15\" class=\"Section3\"\u003e\n\u003ch2\u003eROC analysis\u003c/h2\u003e\n\u003cp\u003eThe results of ROC analysis were presented in Tables\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e6\u003c/span\u003e and \u003cspan class=\"InternalRef\"\u003e7\u003c/span\u003e.\u003c/p\u003e\n\u003cp\u003eThe majority of analyzed cytokines significantly differentiated the patients with ischemic stroke from healthy controls. What is worth mentioning, many of given parameters in stimulated saliva, were characterized by AUC\u0026thinsp;=\u0026thinsp;1, sensitivity and specificity reaching up to 100%, e.g. TNF-\u0026alpha;/IL-10 ratio, IL-6/IL-10 ratio, TNF-\u0026alpha; specific amount, and IL-6 concentration. In non-stimulated saliva, TNF-\u0026alpha;/IL-10 and IL-6/IL-10 ratios distinguished themselves with high AUC and both great sensitivity and specificity.\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec16\" class=\"Section3\"\u003e\n\u003ch2\u003eTNF-\u0026alpha; diagnostic significance\u003c/h2\u003e\n\u003cp\u003eSalivary TNF correlates remarkably negatively with cognitive function in the ACE III scale. Therefore, we examined how this parameter differentiates stroke patients according to cognitive impairment. For this purpose, stroke patients were divided into 3 groups: normal cognitive function (100\u0026thinsp;\u0026minus;\u0026thinsp;89 in the ACE III scale), mild dementia (88\u0026thinsp;\u0026minus;\u0026thinsp;61 in the ACE III scale), and moderate dementia (\u0026lt;\u0026thinsp;61 in the ACE III scale) \u003csup\u003e\u003cspan class=\"CitationRef\"\u003e52\u003c/span\u003e,\u003cspan class=\"CitationRef\"\u003e53\u003c/span\u003e\u003c/sup\u003e. Salivary TNF-\u0026alpha; differentiates (with high sensitivity and specificity) stroke patients with normal cognitive function from patients with mild to moderate dementia (Fig.\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e1\u003c/span\u003e).\u003c/p\u003e\n\u003ctable border=\"1\" width=\"0\"\u003e\u003ccaption\u003e\n\u003cp\u003eTable 4\u003c/p\u003e\n\u003cp\u003eSalivary cytokines in stroke patients and healthy controls.\u003c/p\u003e\n\u003c/caption\u003e\n\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd colspan=\"2\" rowspan=\"3\" width=\"180\"\u003e\n\u003cp\u003e\u003cstrong\u003eSalivary biomarker\u003c/strong\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd colspan=\"5\" width=\"439\"\u003e\n\u003cp\u003e\u003cstrong\u003eNWS\u003c/strong\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd colspan=\"5\" width=\"449\"\u003e\n\u003cp\u003e\u003cstrong\u003eSWS\u003c/strong\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd colspan=\"2\" width=\"184\"\u003e\n\u003cp\u003e\u003cstrong\u003eControl\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e\u003cem\u003en\u003c/em\u003e\u003c/strong\u003e\u003cstrong\u003e = 25\u003c/strong\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd colspan=\"2\" width=\"198\"\u003e\n\u003cp\u003e\u003cstrong\u003eStroke\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e\u003cem\u003en\u003c/em\u003e\u003c/strong\u003e\u003cstrong\u003e = 25\u003c/strong\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd rowspan=\"2\" width=\"57\"\u003e\n\u003cp\u003e\u003cstrong\u003ep value\u003c/strong\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd colspan=\"2\" width=\"184\"\u003e\n\u003cp\u003e\u003cstrong\u003eControl\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e\u003cem\u003en\u003c/em\u003e\u003c/strong\u003e\u003cstrong\u003e = 25\u003c/strong\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd colspan=\"2\" width=\"198\"\u003e\n\u003cp\u003e\u003cstrong\u003eStroke\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e\u003cem\u003en\u003c/em\u003e\u003c/strong\u003e\u003cstrong\u003e = 25\u003c/strong\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd rowspan=\"2\" width=\"66\"\u003e\n\u003cp\u003e\u003cstrong\u003ep value\u003c/strong\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"90\"\u003e\n\u003cp\u003eMean \u0026plusmn; SD\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"94\"\u003e\n\u003cp\u003e95%CI\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"103\"\u003e\n\u003cp\u003eMean \u0026plusmn; SD\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"95\"\u003e\n\u003cp\u003e95%CI\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"97\"\u003e\n\u003cp\u003eMean \u0026plusmn; SD\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"87\"\u003e\n\u003cp\u003e95%CI\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"104\"\u003e\n\u003cp\u003eMean \u0026plusmn; SD\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"94\"\u003e\n\u003cp\u003e95%CI\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd rowspan=\"3\" width=\"85\"\u003e\n\u003cp\u003eTNF-\u0026alpha;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"95\"\u003e\n\u003cp\u003econcentration (pg/mL)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"90\"\u003e\n\u003cp\u003e11.72\u0026plusmn;3.503\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"94\"\u003e\n\u003cp\u003e10.27-13.16\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"103\"\u003e\n\u003cp\u003e27.93\u0026plusmn;6.021\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"95\"\u003e\n\u003cp\u003e25.44-30.41\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"57\"\u003e\n\u003cp\u003e\u0026lt;0.0001\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"97\"\u003e\n\u003cp\u003e4.02\u0026plusmn;0.6272\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"87\"\u003e\n\u003cp\u003e3.761-4.279\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"104\"\u003e\n\u003cp\u003e6.776\u0026plusmn;0.8391\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"94\"\u003e\n\u003cp\u003e6.43-7.122\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"66\"\u003e\n\u003cp\u003e\u0026lt;0.0001\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"95\"\u003e\n\u003cp\u003eoutput\u003c/p\u003e\n\u003cp\u003e(pg/min)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"90\"\u003e\n\u003cp\u003e3.731\u0026plusmn;1.181\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"94\"\u003e\n\u003cp\u003e3.244-4.218\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"103\"\u003e\n\u003cp\u003e11.07\u0026plusmn;7.472\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"95\"\u003e\n\u003cp\u003e7.989-14.16\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"57\"\u003e\n\u003cp\u003e\u0026lt;0.0001\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"97\"\u003e\n\u003cp\u003e3.649\u0026plusmn;1.189\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"87\"\u003e\n\u003cp\u003e3.158-4.14\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"104\"\u003e\n\u003cp\u003e3.947\u0026plusmn;1.768\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"94\"\u003e\n\u003cp\u003e3.217-4.677\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"66\"\u003e\n\u003cp\u003e0.4884\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"95\"\u003e\n\u003cp\u003especific amount (ng/mg protein)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"90\"\u003e\n\u003cp\u003e9.846\u0026plusmn;3.591\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"94\"\u003e\n\u003cp\u003e8.364-11.33\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"103\"\u003e\n\u003cp\u003e26.63\u0026plusmn;11.21\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"95\"\u003e\n\u003cp\u003e22-31.25\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"57\"\u003e\n\u003cp\u003e\u0026lt;0.0001\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"97\"\u003e\n\u003cp\u003e3.106\u0026plusmn;0.5689\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"87\"\u003e\n\u003cp\u003e2.871-3.341\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"104\"\u003e\n\u003cp\u003e7.702\u0026plusmn;2.53\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"94\"\u003e\n\u003cp\u003e6.658-8.746\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"66\"\u003e\n\u003cp\u003e\u0026lt;0.0001\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd rowspan=\"3\" width=\"85\"\u003e\n\u003cp\u003eIL-6\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"95\"\u003e\n\u003cp\u003econcentration (pg/mL)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"90\"\u003e\n\u003cp\u003e15.81\u0026plusmn;3.715\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"94\"\u003e\n\u003cp\u003e14.28-17.34\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"103\"\u003e\n\u003cp\u003e29.27\u0026plusmn;9.684\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"95\"\u003e\n\u003cp\u003e25.28-33.27\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"57\"\u003e\n\u003cp\u003e\u0026lt;0.0001\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"97\"\u003e\n\u003cp\u003e6.996\u0026plusmn;0.8605\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"87\"\u003e\n\u003cp\u003e6.64-7.351\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"104\"\u003e\n\u003cp\u003e11.3\u0026plusmn;1.065\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"94\"\u003e\n\u003cp\u003e10.86-11.74\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"66\"\u003e\n\u003cp\u003e\u0026lt;0.0001\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"95\"\u003e\n\u003cp\u003eoutput\u003c/p\u003e\n\u003cp\u003e(pg/min)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"90\"\u003e\n\u003cp\u003e5.213\u0026plusmn;1.863\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"94\"\u003e\n\u003cp\u003e4.443-5.982\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"103\"\u003e\n\u003cp\u003e11.68\u0026plusmn;8.251\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"95\"\u003e\n\u003cp\u003e8.27-15.08\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"57\"\u003e\n\u003cp\u003e0.0004\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"97\"\u003e\n\u003cp\u003e6.37\u0026plusmn;2.004\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"87\"\u003e\n\u003cp\u003e5.543-7.197\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"104\"\u003e\n\u003cp\u003e6.487\u0026plusmn;2.655\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"94\"\u003e\n\u003cp\u003e5.391-7.583\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"66\"\u003e\n\u003cp\u003e0.8614\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"95\"\u003e\n\u003cp\u003especific amount (ng/mg protein)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"90\"\u003e\n\u003cp\u003e13.27\u0026plusmn;3.674\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"94\"\u003e\n\u003cp\u003e11.75-14.79\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"103\"\u003e\n\u003cp\u003e26.89\u0026plusmn;9.536\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"95\"\u003e\n\u003cp\u003e22.95-30.82\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"57\"\u003e\n\u003cp\u003e\u0026lt;0.0001\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"97\"\u003e\n\u003cp\u003e5.449\u0026plusmn;1.094\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"87\"\u003e\n\u003cp\u003e4.997-5.9\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"104\"\u003e\n\u003cp\u003e12.8\u0026plusmn;3.838\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"94\"\u003e\n\u003cp\u003e11.21-14.38\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"66\"\u003e\n\u003cp\u003e\u0026lt;0.0001\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd rowspan=\"3\" width=\"85\"\u003e\n\u003cp\u003eIL-10\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"95\"\u003e\n\u003cp\u003econcentration (pg/mL)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"90\"\u003e\n\u003cp\u003e3.934\u0026plusmn;0.7284\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"94\"\u003e\n\u003cp\u003e3.634-4.235\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"103\"\u003e\n\u003cp\u003e2.078\u0026plusmn;0.8456\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"95\"\u003e\n\u003cp\u003e1.728-2.427\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"57\"\u003e\n\u003cp\u003e\u0026lt;0.000\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"97\"\u003e\n\u003cp\u003e0.5012\u0026plusmn;0.07068\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"87\"\u003e\n\u003cp\u003e0.4721-0.5304\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"104\"\u003e\n\u003cp\u003e0.1753\u0026plusmn;0.085\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"94\"\u003e\n\u003cp\u003e0.1402-0.2104\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"66\"\u003e\n\u003cp\u003e\u0026lt;0.0001\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"95\"\u003e\n\u003cp\u003eoutput\u003c/p\u003e\n\u003cp\u003e(pg/min)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"90\"\u003e\n\u003cp\u003e0.4502\u0026plusmn;0.1308\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"94\"\u003e\n\u003cp\u003e0.3962-0.5042\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"103\"\u003e\n\u003cp\u003e0.09977\u0026plusmn;0.05173\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"95\"\u003e\n\u003cp\u003e0.07842-0.1211\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"57\"\u003e\n\u003cp\u003e0.0031\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"97\"\u003e\n\u003cp\u003e1.287\u0026plusmn;0.3666\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"87\"\u003e\n\u003cp\u003e1.136-1.438\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"104\"\u003e\n\u003cp\u003e0.8238\u0026plusmn;0.6476\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"94\"\u003e\n\u003cp\u003e0.5564-1.091\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"66\"\u003e\n\u003cp\u003e\u0026lt;0.0001\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"95\"\u003e\n\u003cp\u003especific amount (ng/mg protein)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"90\"\u003e\n\u003cp\u003e3.303\u0026plusmn;0.8475\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"94\"\u003e\n\u003cp\u003e2.953-3.653\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"103\"\u003e\n\u003cp\u003e1.927\u0026plusmn;0.9295\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"95\"\u003e\n\u003cp\u003e1.543-2.31\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"57\"\u003e\n\u003cp\u003e\u0026lt;0.0001\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"97\"\u003e\n\u003cp\u003e0.3881\u0026plusmn;0.07196\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"87\"\u003e\n\u003cp\u003e0.3584-0.4178\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"104\"\u003e\n\u003cp\u003e0.2015\u0026plusmn;0.1105\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"94\"\u003e\n\u003cp\u003e0.1559-0.2471\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"66\"\u003e\n\u003cp\u003e\u0026lt;0.0001\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"85\"\u003e\n\u003cp\u003eTNF-\u0026alpha;/IL-10\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"95\"\u003e\n\u003cp\u003eratio\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"90\"\u003e\n\u003cp\u003e3.119\u0026plusmn;1.285\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"94\"\u003e\n\u003cp\u003e2.588-3.649\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"103\"\u003e\n\u003cp\u003e18.96\u0026plusmn;17.54\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"95\"\u003e\n\u003cp\u003e11.71-26.2\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"57\"\u003e\n\u003cp\u003e\u0026lt;0.0001\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"97\"\u003e\n\u003cp\u003e8.186\u0026plusmn;1.776\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"87\"\u003e\n\u003cp\u003e7.452-8.919\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"104\"\u003e\n\u003cp\u003e58.24\u0026plusmn;56.44\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"94\"\u003e\n\u003cp\u003e34.94-81.54\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"66\"\u003e\n\u003cp\u003e\u0026lt;0.0001\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"85\"\u003e\n\u003cp\u003eIL-6/IL-10\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"95\"\u003e\n\u003cp\u003eratio\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"90\"\u003e\n\u003cp\u003e4.156\u0026plusmn;1.28\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"94\"\u003e\n\u003cp\u003e3.627-4.684\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"103\"\u003e\n\u003cp\u003e19.88\u0026plusmn;22.59\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"95\"\u003e\n\u003cp\u003e10.56-29.21\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"57\"\u003e\n\u003cp\u003e0.0011\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"97\"\u003e\n\u003cp\u003e14.33\u0026plusmn;3.247\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"87\"\u003e\n\u003cp\u003e12.99-15.67\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"104\"\u003e\n\u003cp\u003e95.8\u0026plusmn;86.71\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"94\"\u003e\n\u003cp\u003e60.01-131.6\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"66\"\u003e\n\u003cp\u003e\u0026lt;0.0001\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003eAbbreviations: TNF-\u0026alpha; - tumor necrosis factor \u0026alpha;; IL-6 \u0026ndash; interleukin 6; IL-10 \u0026ndash; interleukin 10\u003c/p\u003e\n\u003cdiv class=\"gridtable\"\u003e\n\u003ctable id=\"Tab4\" border=\"1\"\u003e\u003ccaption\u003e\n\u003cdiv class=\"CaptionNumber\"\u003eTable 5\u003c/div\u003e\n\u003cdiv class=\"CaptionContent\"\u003e\n\u003cp\u003eStatistically significant correlations in patients with ischemic stroke.\u003c/p\u003e\n\u003c/div\u003e\n\u003c/caption\u003e\n\u003cthead\u003e\n\u003ctr\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003ePair of variables\u003c/p\u003e\n\u003c/th\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003er\u003c/p\u003e\n\u003c/th\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003ep value\u003c/p\u003e\n\u003c/th\u003e\n\u003c/tr\u003e\n\u003c/thead\u003e\n\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd colspan=\"3\" align=\"left\"\u003e\n\u003cp\u003e\u003cem\u003eNWS\u003c/em\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eTNF-\u0026alpha; concentration and BBS\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-0.612\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.001\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eTNF-\u0026alpha; concentration and BI\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-0.789\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e\u0026lt;\u0026thinsp;0.001\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eTNF-\u0026alpha; concentration and ACE III\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-0.832\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e\u0026lt;\u0026thinsp;0.001\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eTNF-\u0026alpha; output and BBS\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-0.398\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.045\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eIL-6 concentration and BI\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.464\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.02\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eIL-6 output and FIM\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-0.41\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.042\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eIL-6 specific amount and PlI\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-0.481\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.015\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eIL-6 specific amount and BI\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.418\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.037\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd colspan=\"3\" align=\"left\"\u003e\n\u003cp\u003e\u003cem\u003eSWS\u003c/em\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eTNF-\u0026alpha; output and BBS\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-0.474\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.017\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eTNF-\u0026alpha; output and BI\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-0.761\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e\u0026lt;\u0026thinsp;0.001\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eTNF-\u0026alpha; output and ACE III\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-0.801\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e\u0026lt;\u0026thinsp;0.001\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eIL-6 output and BBS\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-0.512\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.009\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eIL-6 output and BI\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-0.796\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e\u0026lt;\u0026thinsp;0.001\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eIL-6 output and ACE III\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-0.754\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e\u0026lt;\u0026thinsp;0.001\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003eAbbreviations: NWS \u0026ndash; non-stimulated whole saliva; SWS \u0026ndash; stimulated whole saliva; TNF-\u0026alpha; - tumor necrosis factor \u0026alpha;; IL-6 \u0026ndash; interleukin 6; PLI \u0026ndash; Plaque Index; ACE III \u0026ndash; Addenbrooke\u0026rsquo;s Cognitive Examination; FIM \u0026ndash; functional independence measure; BBS \u0026ndash; Berg Balance Scale;\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv class=\"gridtable\"\u003e\n\u003ctable id=\"Tab6\" border=\"1\"\u003e\u003ccaption\u003e\n\u003cdiv class=\"CaptionNumber\"\u003eTable 6\u003c/div\u003e\n\u003cdiv class=\"CaptionContent\"\u003e\n\u003cp\u003eReceiver operating characteristics of salivary cytokines in non-stimulated saliva of stroke patients and healthy controls.\u003c/p\u003e\n\u003c/div\u003e\n\u003c/caption\u003e\n\u003cthead\u003e\n\u003ctr\u003e\n\u003cth colspan=\"2\" rowspan=\"2\" align=\"left\"\u003e\n\u003cp\u003eSalivary biomarker\u003c/p\u003e\n\u003c/th\u003e\n\u003cth colspan=\"8\" align=\"left\"\u003e\n\u003cp\u003eNWS\u003c/p\u003e\n\u003c/th\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003e\u003cstrong\u003eAUC\u003c/strong\u003e\u003c/p\u003e\n\u003c/th\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003e\u003cstrong\u003e95% CI\u003c/strong\u003e\u003c/p\u003e\n\u003c/th\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003e\u003cstrong\u003eP-value\u003c/strong\u003e\u003c/p\u003e\n\u003c/th\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003e\u003cstrong\u003eCut off\u003c/strong\u003e\u003c/p\u003e\n\u003c/th\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003e\u003cstrong\u003eSensitivity%\u003c/strong\u003e\u003c/p\u003e\n\u003c/th\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003e\u003cstrong\u003e95% CI\u003c/strong\u003e\u003c/p\u003e\n\u003c/th\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003e\u003cstrong\u003eSpecificity%\u003c/strong\u003e\u003c/p\u003e\n\u003c/th\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003e\u003cstrong\u003e95% CI\u003c/strong\u003e\u003c/p\u003e\n\u003c/th\u003e\n\u003c/tr\u003e\n\u003c/thead\u003e\n\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd rowspan=\"3\" align=\"left\"\u003e\n\u003cp\u003eTNF-\u0026alpha;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003econcentration (pg/mL)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.992\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.9760\u0026ndash;1.000\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e\u0026lt;\u0026thinsp;0.0001\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e\u0026gt;\u0026thinsp;17.46\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e96\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e80.46\u0026ndash;99.79\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e96\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e80.46\u0026ndash;99.79\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eoutput\u0026nbsp;(pg/min)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.8544\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.7470\u0026ndash;0.9618\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e\u0026lt;\u0026thinsp;0.0001\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e\u0026gt;\u0026thinsp;4.727\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e76\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e56.57\u0026ndash;88.50\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e80\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e60.87\u0026ndash;91.14\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003especific amount (ng/mg protein)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.9712\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.9340\u0026ndash;1.000\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e\u0026lt;\u0026thinsp;0.0001\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e\u0026gt;\u0026thinsp;15.61\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e92\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e75.03\u0026ndash;98.58\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e92\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e75.03\u0026ndash;98.58\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd rowspan=\"3\" align=\"left\"\u003e\n\u003cp\u003eIL-6\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003econcentration (pg/mL)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.8976\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.7944\u0026ndash;1.000\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e\u0026lt;\u0026thinsp;0.0001\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e\u0026gt;\u0026thinsp;22.08\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e84\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e65.35\u0026ndash;93.60\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e96\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e80.46\u0026ndash;99.79\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eoutput\u0026nbsp;(pg/min)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.7376\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.5897\u0026ndash;0.8855\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.004\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e\u0026gt;\u0026thinsp;5.602\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e76\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e56.57\u0026ndash;88.50\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e72\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e52.42\u0026ndash;85.72\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003especific amount (ng/mg protein)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.9104\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.8168\u0026ndash;1.000\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e\u0026lt;\u0026thinsp;0.0001\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e\u0026gt;\u0026thinsp;18.03\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e84\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e65.35\u0026ndash;93.60\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e92\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e75.03\u0026ndash;98.58\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd rowspan=\"3\" align=\"left\"\u003e\n\u003cp\u003eIL-10\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003econcentration (pg/mL)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.9568\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.9078\u0026ndash;1.000\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e\u0026lt;\u0026thinsp;0.0001\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e\u0026lt;\u0026thinsp;3.014\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e88\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e70.04\u0026ndash;95.83\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e92\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e75.03\u0026ndash;98.58\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eoutput\u0026nbsp;(pg/min)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.776\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.6400\u0026ndash;0.9120\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.0008\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e\u0026lt;\u0026thinsp;1.140\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e80\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e60.87\u0026ndash;91.14\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e68\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e48.41\u0026ndash;82.79\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003especific amount (ng/mg protein)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.8848\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.7883\u0026ndash;0.9813\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e\u0026lt;\u0026thinsp;0.0001\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e\u0026lt;\u0026thinsp;2.173\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e72\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e52.42\u0026ndash;85.72\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e96\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e80.46\u0026ndash;99.79\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd colspan=\"2\" align=\"left\"\u003e\n\u003cp\u003eTNF-\u0026alpha;/IL-10\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.9984\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.9931\u0026ndash;1.000\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e\u0026lt;\u0026thinsp;0.0001\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e\u0026gt;\u0026thinsp;5.577\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e100\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e86.68\u0026ndash;100.0\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e96\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e80.46\u0026ndash;99.79\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd colspan=\"2\" align=\"left\"\u003e\n\u003cp\u003eIL-6/IL-10\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.9792\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.9426\u0026ndash;1.000\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e\u0026lt;\u0026thinsp;0.0001\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e\u0026gt;\u0026thinsp;7.625\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e92\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e75.03\u0026ndash;98.58\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e100\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e86.68\u0026ndash;100.0\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003eAbbreviations: TNF-\u0026alpha; - tumor necrosis factor \u0026alpha;; IL-6 \u0026ndash; interleukin 6; IL-10 \u0026ndash; interleukin 10.\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv class=\"gridtable\"\u003e\n\u003ctable id=\"Tab7\" border=\"1\"\u003e\u003ccaption\u003e\n\u003cdiv class=\"CaptionNumber\"\u003eTable 7\u003c/div\u003e\n\u003cdiv class=\"CaptionContent\"\u003e\n\u003cp\u003eReceiver operating characteristics of salivary cytokines in stimulated saliva of stroke patients and healthy controls.\u003c/p\u003e\n\u003c/div\u003e\n\u003c/caption\u003e\n\u003cthead\u003e\n\u003ctr\u003e\n\u003cth colspan=\"2\" rowspan=\"2\" align=\"left\"\u003e\n\u003cp\u003eSalivary biomarker\u003c/p\u003e\n\u003c/th\u003e\n\u003cth colspan=\"8\" align=\"left\"\u003e\n\u003cp\u003eSWS\u003c/p\u003e\n\u003c/th\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003e\u003cstrong\u003eAUC\u003c/strong\u003e\u003c/p\u003e\n\u003c/th\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003e\u003cstrong\u003e95% CI\u003c/strong\u003e\u003c/p\u003e\n\u003c/th\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003e\u003cstrong\u003eP-value\u003c/strong\u003e\u003c/p\u003e\n\u003c/th\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003e\u003cstrong\u003eCut off\u003c/strong\u003e\u003c/p\u003e\n\u003c/th\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003e\u003cstrong\u003eSensitivity%\u003c/strong\u003e\u003c/p\u003e\n\u003c/th\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003e\u003cstrong\u003e95% CI\u003c/strong\u003e\u003c/p\u003e\n\u003c/th\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003e\u003cstrong\u003eSpecificity%\u003c/strong\u003e\u003c/p\u003e\n\u003c/th\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003e\u003cstrong\u003e95% CI\u003c/strong\u003e\u003c/p\u003e\n\u003c/th\u003e\n\u003c/tr\u003e\n\u003c/thead\u003e\n\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd rowspan=\"3\" align=\"left\"\u003e\n\u003cp\u003eTNF-\u0026alpha;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003econcentration (pg/mL)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.9984\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.9931 to 1.000\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e\u0026lt;\u0026thinsp;0.0001\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e\u0026gt;\u0026thinsp;4.941\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e100\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e86.68\u0026ndash;100.0\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e96\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e80.46\u0026ndash;99.79\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eoutput\u0026nbsp;(pg/min)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.5616\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.3950 to 0.7282\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.4551\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e\u0026gt;\u0026thinsp;4.211\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e52\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e33.50\u0026ndash;69.97\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e76\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e56.57\u0026ndash;88.50\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003especific amount (ng/mg protein)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e1\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e1.000\u0026ndash;1.000\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e\u0026lt;\u0026thinsp;0.0001\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e\u0026gt;\u0026thinsp;4.249\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e100\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e86.68\u0026ndash;100.0\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e100\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e86.68\u0026ndash;100.0\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd rowspan=\"3\" align=\"left\"\u003e\n\u003cp\u003eIL-6\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003econcentration (pg/mL)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e1\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e1.000\u0026ndash;1.000\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e\u0026lt;\u0026thinsp;0.0001\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e\u0026gt;\u0026thinsp;8.986\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e100\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e86.68\u0026ndash;100.0\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e100\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e86.68\u0026ndash;100.0\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eoutput\u0026nbsp;(pg/min)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.536\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.3704\u0026ndash;0.7016\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.6624\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e\u0026gt;\u0026thinsp;6.640\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e60\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e40.74\u0026ndash;76.60\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e60\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e40.74\u0026ndash;76.60\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003especific amount (ng/mg protein)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.9968\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.9882\u0026ndash;1.000\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e\u0026lt;\u0026thinsp;0.0001\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e\u0026gt;\u0026thinsp;8.366\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e96\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e80.46\u0026ndash;99.79\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e100\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e86.68\u0026ndash;100.0\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd rowspan=\"3\" align=\"left\"\u003e\n\u003cp\u003eIL-10\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003econcentration (pg/mL)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.9984\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.9931\u0026ndash;1.000\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e\u0026lt;\u0026thinsp;0.0001\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e\u0026lt;\u0026thinsp;0.3165\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e96\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e80.46\u0026ndash;99.79\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e100\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e86.68\u0026ndash;100.0\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eoutput\u0026nbsp;(pg/min)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e1\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e1.000\u0026ndash;1.000\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e\u0026lt;\u0026thinsp;0.0001\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e\u0026lt;\u0026thinsp;0.2250\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e100\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e86.68\u0026ndash;100.0\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e100\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e86.68\u0026ndash;100.0\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003especific amount (ng/mg protein)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.9136\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.8353\u0026ndash;0.9919\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e\u0026lt;\u0026thinsp;0.0001\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e\u0026lt;\u0026thinsp;0.2665\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e76\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e56.57\u0026ndash;88.50\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e96\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e80.46\u0026ndash;99.79\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd colspan=\"2\" align=\"left\"\u003e\n\u003cp\u003eTNF-\u0026alpha;/IL-10\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e1\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e1.000\u0026ndash;1.000\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e\u0026lt;\u0026thinsp;0.0001\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e\u0026gt;\u0026thinsp;17.01\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e100\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e86.68\u0026ndash;100.0\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e100\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e86.68\u0026ndash;100.0\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd colspan=\"2\" align=\"left\"\u003e\n\u003cp\u003eIL-6/IL-10\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e1\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e1.000\u0026ndash;1.000\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e\u0026lt;\u0026thinsp;0.0001\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e\u0026gt;\u0026thinsp;24.60\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e100\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e86.68\u0026ndash;100.0\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e100\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e86.68\u0026ndash;100.0\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003c/tbody\u003e\n\u003c/table\u003e\n\u003c/div\u003e\n\u003cp\u003eAbbreviations: TNF-\u0026alpha; - tumor necrosis factor \u0026alpha;; IL-6 \u0026ndash; interleukin 6; IL-10 \u0026ndash; interleukin 10.\u003c/p\u003e\n\u003c/div\u003e\n\u003c/div\u003e"},{"header":"Discussion","content":"\u003cp\u003eOur study is the first to evaluate the salivary cytokine profile in patients with ischemic stroke. We have demonstrated that assessment of TNF-\u0026alpha;, IL-6, and IL-10 levels in both NWS and SWS significantly differentiates stroke patients from healthy controls. However, particular attention should be paid to salivary TNF-\u0026alpha;, which may be a non-invasive biomarker of cognitive function/physical status in stroke subjects.\u003c/p\u003e\n\u003cp\u003eIn recent years, increasing attention has been given to the potential use of saliva in laboratory diagnostics. This biomaterial allows for repeated, non-invasive, and stress-free sampling, which is especially important in disabled and elderly patients \u003csup\u003e\u003cspan class=\"CitationRef\"\u003e54\u003c/span\u003e\u003c/sup\u003e. Saliva is also a non-infectious and relatively durable bioliquid \u003csup\u003e\u003cspan class=\"CitationRef\"\u003e29\u003c/span\u003e,\u003cspan class=\"CitationRef\"\u003e55\u003c/span\u003e\u003c/sup\u003e. The concentration of many biomarkers in saliva highly correlates with their blood/CSF levels, allowing saliva to be used in the diagnosis of various systemic diseases \u003csup\u003e\u003cspan class=\"CitationRef\"\u003e56\u003c/span\u003e\u0026ndash;\u003cspan class=\"CitationRef\"\u003e59\u003c/span\u003e\u003c/sup\u003e. Indeed, saliva can be an alternative to blood for the determination of enzymes, hormones, immunoglobulins, and redox biomarkers \u003csup\u003e\u003cspan class=\"CitationRef\"\u003e60\u003c/span\u003e,\u003cspan class=\"CitationRef\"\u003e61\u003c/span\u003e\u003c/sup\u003e. Saliva has also been shown to be valuable in assessing cytokines, chemokines, pro-inflammatory enzymes, and growth factors \u003csup\u003e\u003cspan class=\"CitationRef\"\u003e62\u003c/span\u003e\u0026ndash;\u003cspan class=\"CitationRef\"\u003e64\u003c/span\u003e\u003c/sup\u003e. Although salivary cytokines' diagnostic utility has been demonstrated in metabolic and neurodegenerative diseases \u003csup\u003e\u003cspan class=\"CitationRef\"\u003e35\u003c/span\u003e\u0026ndash;\u003cspan class=\"CitationRef\"\u003e39\u003c/span\u003e\u003c/sup\u003e, there are no data on ischemic stroke patients.\u003c/p\u003e\n\u003cp\u003eWe showed that salivary TNF-\u0026alpha; and IL-6 were significantly higher, whereas IL-10 content was statistically lower in ischemic stroke patients than healthy controls. TNF-\u0026alpha;/IL-10 and IL-6/IL-10 ratio were also significantly higher in NWS and SWS of stroke patients, indicating an intensification of the inflammatory process and a limited anti-inflammatory role of IL-10. As we have shown by ROC analysis, salivary biomarkers are also characterized by a high diagnostic utility in stroke diagnostics. Salivary TNF-\u0026alpha;, IL-6, and IL-10 as well as TNF-\u0026alpha;/IL-10 and IL-6/IL-10 ratio significantly distinguish stroke patients from controls with high sensitivity and specificity. This fact should not be surprising because inflammatory and immune factors have been indicated as potential stroke biomarkers \u003csup\u003e\u003cspan class=\"CitationRef\"\u003e65\u003c/span\u003e,\u003cspan class=\"CitationRef\"\u003e66\u003c/span\u003e\u003c/sup\u003e. Indeed, in stroke patients, activation of CNS cells (mainly microglia and astrocytes) and increased expression of blood cytokines, interleukins, chemokines, adhesion molecules, and pro-inflammatory enzymes is observed \u003csup\u003e\u003cspan class=\"CitationRef\"\u003e67\u003c/span\u003e\u003c/sup\u003e. Interestingly, the severity of inflammation correlates with brain infarct size and neurological deficits in CT and physical examination \u003csup\u003e68\u0026minus;70\u003c/sup\u003e. However, cytokines' diagnostic utility has only been demonstrated in plasma/serum and CSF of ischemic stroke patients \u003csup\u003e70\u0026minus;72\u003c/sup\u003e. Our study is the first to show the utility of saliva to determine cytokine levels in stroke subjects. This raises hopes for non-invasive, painless, and non-stressful stroke diagnostics. It is believed that cytokines synthesized in the stroke focus pass through the damaged BBB into the blood and then into saliva by passive transport (diffusion or ultrafiltration) or by a specific transporter (facilitated diffusion or active transport) \u003csup\u003e\u003cspan class=\"CitationRef\"\u003e73\u003c/span\u003e,\u003cspan class=\"CitationRef\"\u003e74\u003c/span\u003e\u003c/sup\u003e. The BBB has a prominent role in the development of the inflammatory response by increasing the cytokine circulation between the CSF and the bloodstream \u003csup\u003e\u003cspan class=\"CitationRef\"\u003e9\u003c/span\u003e\u003c/sup\u003e. Unfortunately, in our research, we did not assess the cytokine content in blood, so further studies are needed to evaluate stroke patients' saliva-blood relationship. Nevertheless, it can be speculated that, as in other CNS diseases, salivary cytokine concentrations reflect their plasma/CSF levels \u003csup\u003e\u003cspan class=\"CitationRef\"\u003e25\u003c/span\u003e,\u003cspan class=\"CitationRef\"\u003e28\u003c/span\u003e,\u003cspan class=\"CitationRef\"\u003e75\u003c/span\u003e\u003c/sup\u003e.\u003c/p\u003e\n\u003cp\u003eIn our patients, the content of pro-inflammatory TNF-\u0026alpha; and IL-6 was higher, and anti-inflammatory IL-10 was lower in both NWS and SWS of stroke patients. However, it is important to note that saliva's cytokine profile depends on the type of salivary gland involved in saliva secretion. Indeed, up to 70% of non-stimulated saliva is produced in the submandibular glands, making the NWS composition very similar to blood plasma. During stimulation, the parotid gland's contribution to saliva formation increases \u003csup\u003e\u003cspan class=\"CitationRef\"\u003e76\u003c/span\u003e\u003c/sup\u003e. Stimulated saliva is characterized by significantly higher concentrations of Na\u003csup\u003e+\u003c/sup\u003e, HCO\u003csub\u003e3\u003c/sub\u003e\u003csup\u003e\u0026minus;\u003c/sup\u003e and Cl\u003csup\u003e\u0026minus;\u003c/sup\u003e ions and lower contents of K\u003csup\u003e+\u003c/sup\u003e and phosphate ions and mucins compared to non-stimulated saliva \u003csup\u003e\u003cspan class=\"CitationRef\"\u003e77\u003c/span\u003e\u003c/sup\u003e. Cytokine production in salivary gland cells also increases, making the composition of SWS more different from that of plasma \u003csup\u003e\u003cspan class=\"CitationRef\"\u003e78\u003c/span\u003e\u003c/sup\u003e. Therefore, evaluation of cytokines in NWS rather than in SWS may be of greater diagnostic value. In our study, among all assessed cytokines, tumor necrosis factor \u0026alpha; deserves special attention. Salivary TNF-\u0026alpha; differentiates stroke patients from controls with high sensitivity and specificity and correlates with cognitive dysfunction in the ACE III scale and severity of functional impairment in the BBS and ADL scale. Therefore, in the next step, we examined how this parameter distinguishes stroke patients according to cognitive impairment. For this purpose, stroke subjects were divided into three groups: normal cognition (100\u0026thinsp;\u0026minus;\u0026thinsp;89 in the ACE III scale), mild dementia (88\u0026thinsp;\u0026minus;\u0026thinsp;61 in the ACE III scale), and moderate dementia (\u0026lt;\u0026thinsp;61 in the ACE III scale) \u003csup\u003e\u003cspan class=\"CitationRef\"\u003e52\u003c/span\u003e,\u003cspan class=\"CitationRef\"\u003e53\u003c/span\u003e\u003c/sup\u003e. Salivary TNF-\u0026alpha; differentiates (with high sensitivity and specificity) stroke patients with normal cognitive function from patients with mild to moderate dementia (Fig.\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e1\u003c/span\u003e). Although further studies in a larger patient population are needed, TNF-\u0026alpha; may be a potential biomarker of cognitive impairment/functional status in stroke cases. As shown in previous studies, elevated TNF-\u0026alpha; serum level was noted already in the first few hours after the ischemic stroke \u003csup\u003e\u003cspan class=\"CitationRef\"\u003e10\u003c/span\u003e,\u003cspan class=\"CitationRef\"\u003e16\u003c/span\u003e\u003c/sup\u003e. Serum TNF-\u0026alpha; also correlates with brain infarct volume and severity of neurological symptoms as well as reflects the degree of patients' functional disability \u003csup\u003e\u003cspan class=\"CitationRef\"\u003e79\u003c/span\u003e,\u003cspan class=\"CitationRef\"\u003e80\u003c/span\u003e\u003c/sup\u003e. Indeed, TNF-\u0026alpha; has a unique role in stroke pathogenesis. Primarily, it is the first inflammatory mediator produced in the ischemic area \u003csup\u003e\u003cspan class=\"CitationRef\"\u003e6\u003c/span\u003e,\u003cspan class=\"CitationRef\"\u003e80\u003c/span\u003e\u003c/sup\u003e. Secondly, it stimulates the synthesis/secretion of chemotactic factors that stimulate peripheral blood leukocytes and activates other cytokines like IL-1, IL-6, and interferon-gamma (IFN-\u0026gamma;) \u003csup\u003e\u003cspan class=\"CitationRef\"\u003e10\u003c/span\u003e\u003c/sup\u003e. TNF-\u0026alpha; also enhances MMPs activity \u003csup\u003e\u003cspan class=\"CitationRef\"\u003e81\u003c/span\u003e\u003c/sup\u003e, thereby increasing endothelial cell permeability and BBB dysfunction \u003csup\u003e\u003cspan class=\"CitationRef\"\u003e82\u003c/span\u003e\u003c/sup\u003e. Our study is the first to demonstrate that salivary TNF-\u0026alpha; can be a potential biomarker of stroke patient\u0026rsquo;s functioning. Interestingly, the level of TNF-\u0026alpha; in NWS correlates more strongly with the cognitive and physical functional status of stroke patients. Thus, non-stimulated saliva has a better diagnostic potential than SWS.\u003c/p\u003e\n\u003cp\u003eDespite the unquestionable advantages, the use of saliva in diagnostics also has some drawbacks. The biomarker concentration in saliva may be highly dependent on the degree of salivary gland secretion \u003csup\u003e\u003cspan class=\"CitationRef\"\u003e83\u003c/span\u003e\u003c/sup\u003e. Because salivary hypofunction may occur in stroke patients \u003csup\u003e\u003cspan class=\"CitationRef\"\u003e84\u003c/span\u003e,\u003cspan class=\"CitationRef\"\u003e85\u003c/span\u003e\u003c/sup\u003e, in our study, the concentration of inflammatory biomarkers was standardized to total protein content and salivary flow rate. Nevertheless, both the concentration (pg/mL), salivary output (pg/min), and specific content (per mg protein) of TNF-\u0026alpha; and IL-6 were significantly higher, whereas IL-10 was statistically lower in NWS and SWS of stroke patients compared to controls. Importantly, we did not observe a relationship between salivary gland function and the assessed cytokines. This suggests a negligible effect of salivary hypofunction on salivary TNF-\u0026alpha;, IL-6, and IL-10 levels. Nevertheless, aging, medications, diet supplements, smoking, as well as oral and systemic diseases can also modify the cytokine profile of saliva \u003csup\u003e\u003cspan class=\"CitationRef\"\u003e86\u003c/span\u003e\u0026ndash;\u003cspan class=\"CitationRef\"\u003e88\u003c/span\u003e\u003c/sup\u003e. At this time, it is important to emphasize the careful selection of the control group in the presented study. Control individuals are matched not only by age and sex to the study group but also by other clinical features, including concomitant diseases, medications, oral hygiene status, etc. This significantly reduces the impact of confounding factors on the obtained results.\u003c/p\u003e\n\u003cp\u003eIn summary, stroke patients have increased salivary secretion of pro-inflammatory cytokines and decreased anti-inflammatory mediators. Salivary TNF-\u0026alpha;, IL-6, and IL-10 may be potential non-invasive biomarkers that distinguish stroke patients from controls with high sensitivity and specificity. Of particular note is salivary TNF-\u0026alpha;, which may indicate cognitive/physical impairment in post-stroke individuals. Further studies are needed in a larger stroke population to assess cytokines at both the salivary and central (plasma/serum) levels. Evaluation of other inflammatory mediators for their potential diagnostic significance also requires research.\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eAuthor Contributions: \u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eStudy concept and design: M.M. and K.G. Preparation of dental examination: P.G., K.G., and A.Z. Carrying out dental examination: P.G., K.G., and A.Z. Carrying out general examination: P.G and K.H. Collection of literature data: M.M and K.G. Analysis and interpretation of epidemiological data: M.M., K.G, and A.Z. Sample preparation: M.M and K.M.M. Laboratory determinations: M.M and K.M.M. Statistical analysis: M.M. Drafting of the manuscript: M.M, K.M.M. and K.G. Critical revision of the manuscript for important intellectual content: M.M., P.G., K.G., A.Z., and K.H. Study supervision: M.M., K.G. and A.Z. All authors approved the manuscript. All authors have read and agreed to the published version of the manuscript.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFunding:\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThis work was granted by the Medical University of Bialystok, Poland (grant number: SUB/1/DN/21/002/3330, SUB/1/DN/21/002/1209. Dr. Mateusz Maciejczyk was supported by the Foundation for Polish Science (FNP).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eInstitutional Review Board Statement:\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe study was conducted according to the guidelines of the Declaration of Helsinki, and approved by the Ethics Committee of the Poznan University of Medical Sciences (resolutions 59/19 and 890/19) in Poland.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eInformed Consent Statement:\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eInformed consent was obtained from all subjects involved in the study.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eData Availability Statement:\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe datasets generated for this study are available on request to the corresponding author.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAcknowledgments: \u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe authors would like to thank the head and staff of the health care center (Bonifraterskie Centrum Zdrowia) in Piaski\u0026ndash;Marysin (Piaski, Poland) for consent and help in organization of examination of the patients and all patients who participated in the research for their consent and cooperation. In addition, we would like to express our gratitude to neuropsychologist, Mrs. Hanna Norman, for her help in cognitive assessment of the patients.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConflicts of Interest: \u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe authors declare no conflict of interest.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\n\u003cli\u003eGuzik, A. \u0026amp; Bushnell, C. Stroke Epidemiology and Risk Factor Management. \u003cem\u003eCONTINUUM Lifelong Learning in Neurology\u003c/em\u003e (2017) doi:10.1212/CON.0000000000000416.\u003c/li\u003e\n\u003cli\u003eJayaraj, R. L., Azimullah, S., Beiram, R., Jalal, F. Y. \u0026amp; Rosenberg, G. A. Neuroinflammation: friend and foe for ischemic stroke. \u003cem\u003eJ. Neuroinflammation\u003c/em\u003e \u003cstrong\u003e16\u003c/strong\u003e, 142 (2019).\u003c/li\u003e\n\u003cli\u003eFeigin, V. L., Norrving, B. \u0026amp; Mensah, G. A. Global Burden of Stroke. \u003cem\u003eCirc. 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Salivary markers of kidney function - Potentials and limitations. \u003cem\u003eClinica Chimica Acta\u003c/em\u003e (2016) doi:10.1016/j.cca.2015.11.028.\u003c/li\u003e\n\u003c/ol\u003e"},{"header":"Supplemental materials","content":"\u003cp\u003eSupplementary material file is not available with this version.\u003c/p\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":false,"highlight":"","institution":"","isAcceptedByJournal":true,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"scientific-reports","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"scirep","sideBox":"Learn more about [Scientific Reports](http://www.nature.com/srep/)","snPcode":"","submissionUrl":"","title":"Scientific Reports","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"stoa","reportingPortfolio":"Scientific Reports","inReviewEnabled":true,"inReviewRevisionsEnabled":true},"keywords":"stroke, saliva, inflammation, cytokines, salivary biomarkers, salivary diagnostics","lastPublishedDoi":"10.21203/rs.3.rs-494834/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-494834/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eInflammation plays a crucial role in stroke pathogenesis. Thus, it is not surprising that cytokines, chemokines, and growth factors have been advocated in stroke diagnostics. Our study is the first to evaluate the salivary cytokine profile in patients with ischemic stroke. Twenty-five patients with subacute ischemic stroke and an age-, sex-, and oral hygiene status-matched control group were enrolled in the study. The number of patients was set \u003cem\u003ea priori\u003c/em\u003e based on our previous experiment (α = 0.05, test power = 0.9). Salivary concentrations of tumor necrosis factor α (TNF-α), interleukin 6 (IL-6), and interleukin 10 (IL-10) were assessed using an ELISA method. We showed that salivary TNF-α and IL-6 were significantly higher, whereas IL-10 content was statistically lower in both non-stimulated (NWS) and stimulated (SWS) whole saliva of ischemic stroke patients. However, evaluation of cytokines in NWS rather than in SWS may be of greater diagnostic value. Of particular note is salivary TNF-α, which may indicate cognitive/physical impairment in post-stroke individuals. This parameter distinguishes stroke patients from healthy controls and correlates with cognitive dysfunction and severity of functional impairment. It also differentiates (with high sensitivity and specificity) stroke patients with normal cognition from mild to moderate dementia patients. Saliva may be an alternative to blood for assessing cytokines in stroke patients, although further studies on a larger patient population are needed.\u003c/p\u003e","manuscriptTitle":"Salivary Cytokine Profile in Patients with Ischemic Stroke. Can Salivary TNF-α be a Non-invasive Biomarker of Cognitive Impairment in Stroke Cases?","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2021-05-07 19:09:50","doi":"10.21203/rs.3.rs-494834/v1","editorialEvents":[{"type":"communityComments","content":0},{"type":"decision","content":"Major revision","date":"2021-08-03T05:52:18+00:00","index":"","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2021-07-21T13:43:38+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"5792a50a-a57e-4d78-b8b5-d482d579dc99","date":"2021-07-19T16:25:07+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"401677d6-d7da-4b2c-9d5f-a4967a81e4e4","date":"2021-07-15T14:32:12+00:00","index":"hide","fulltext":""},{"type":"reviewersInvited","content":"","date":"2021-06-16T14:36:31+00:00","index":"","fulltext":""},{"type":"editorAssigned","content":"","date":"2021-06-16T14:24:40+00:00","index":"","fulltext":""},{"type":"editorInvited","content":"","date":"2021-05-07T09:12:15+00:00","index":"","fulltext":""},{"type":"checksComplete","content":"","date":"2021-05-06T05:15:45+00:00","index":"","fulltext":""},{"type":"submitted","content":"Scientific Reports","date":"2021-05-05T08:26:16+00:00","index":"","fulltext":""}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"scientific-reports","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"scirep","sideBox":"Learn more about [Scientific Reports](http://www.nature.com/srep/)","snPcode":"","submissionUrl":"","title":"Scientific Reports","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"stoa","reportingPortfolio":"Scientific Reports","inReviewEnabled":true,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"402954dd-15e0-4526-aa27-a22713fe5562","owner":[],"postedDate":"May 7th, 2021","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"under-review","subjectAreas":[{"id":4168955,"name":"Cardiac \u0026 Cardiovascular Systems"},{"id":4168956,"name":"Cognitive Neuroscience"}],"tags":[],"updatedAt":"2021-08-17T05:14:09+00:00","versionOfRecord":[],"versionCreatedAt":"2021-05-07 19:09:50","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-494834","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-494834","identity":"rs-494834","version":["v1"]},"buildId":"_2-kVJe1T_tPrBINL-cwx","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}