Antioxidant Activity of Saliva and Periodontal Disease

The antioxidant activity of saliva has been investigated in 28 apparently healthy individuals and seven dental patients with periodontal disease. The results show that the major aqueous antioxidant component of whole saliva is uric acid, with lesser contributions from ascorbic acid and albumin. All are present at lower concentrations than those found in the plasma water. The total antioxidant activity (TAA) of saliva correlates (r² = 0.972) with the concentration of uric acid, which contributes more than 70% of the TAA. Stimulation of salivary flow is associated with increased production of antioxidants. The antioxidant potential of saliva does not appear to be compromised in patients with periodontal disease but this may relate to the antioxidant flow from the gingival crevicular fluid.     

Antioxidant status and dialysis: Plasma and saliva antioxidant activity in patients with fluctuating urate levels

The present study is concerned with the influence of processes occurring during dialysis on the antioxidant capacity of plasma and saliva. The biological fluids were also tested for uric acid and total protein content. Before hemodialysis, plasma antioxidant status of hemodialyzed patients appears slightly higher than the corresponding status in normal subjects; after hemodialysis it is found unchanged. The result can be explained by a balance between a reduction in uric acid plasma content, due to the dialytic procedure, and an increase in protein content, possibly due to a dialysis-related hemoconcentration. Moreover, pre-dialysis total antioxidant capacity of whole saliva samples is higher than in healthy individuals and drastically decreases towards normal values following dialytic procedure. Our data indicate a certain concentration of the uric acid in the saliva of hemodialyzed patients and evidence that both total protein concentration and uric acid level show a good correlation with saliva total antioxidant capacity, suggesting that proteins are major antioxidants of this fluid. Further observations are needed to assess whether this improved saliva antioxidant ability has any consequence on the periodontal conditions of hemodialyzed subjects.     

Antioxidant status and dialysis: Plasma and saliva antioxidant activity in patients with fluctuating urate levels

The present study is concerned with the influence of processes occurring during dialysis on the antioxidant capacity of plasma and saliva. The biological fluids were also tested for uric acid and total protein content. Before hemodialysis, plasma antioxidant status of hemodialyzed patients appears slightly higher than the corresponding status in normal subjects; after hemodialysis it is found unchanged. The result can be explained by a balance between a reduction in uric acid plasma content, due to the dialytic procedure, and an increase in protein content, possibly due to a dialysis-related hemoconcentration. Moreover, pre-dialysis total antioxidant capacity of whole saliva samples is higher than in healthy individuals and drastically decreases towards normal values following dialytic procedure. Our data indicate a certain concentration of the uric acid in the saliva of hemodialyzed patients and evidence that both total protein concentration and uric acid level show a good correlation with saliva total antioxidant capacity, suggesting that proteins are major antioxidants of this fluid. Further observations are needed to assess whether this improved saliva antioxidant ability has any consequence on the periodontal conditions of hemodialyzed subjects.     

Salivary uric acid at the acidic pH of the stomach is the principal defense against nitrite-derived reactive species: sparing effects of chlorogenic acid and serum albumin

A complex antioxidant system is present in human saliva, with uric acid being the most concentrated component. Ascorbic acid, present at low concentrations in saliva, is actively secreted into the gastric lumen. We report that ascorbic acid added to human saliva at pH 2 was consumed within a few minutes, regenerating HNO₂, whereas uric acid was consumed relatively slowly in a nitrite-dependent manner. The consumption of uric acid was (i) rapid under normoxic conditions and slower at low oxygen tensions, (ii) coupled to NO release, (iii) linked to the decrease in nitrite consumption and in nitrate formation, and (iv) unaffected by the nitrosation catalyst thiocyanate. Both chlorogenic acid and bovine serum albumin, representative of a phenol- and a protein-rich meal, respectively, were able to spare uric acid, although chlorogenic acid increased, whereas bovine serum albumin inhibited, NO release. We hypothesize that the major role of uric acid in saliva at pH 2 could be to preserve the stomach from the formation of toxic nitrogen species and that low levels of uric acid, together with ascorbic acid consumption, may contribute to the high occurrence of tumors at the gastroesophageal junction and cardia. The sparing effects of dietary compounds may therefore be an important not fully appreciated effect.     

Total reactive antioxidant potential in human saliva of smokers and non-smokers.

Uric acid is the most important non-enzymatic antioxidant present in human saliva. There is a great variability among individuals, both in salivary uric acid content and saliva total reactive antioxidant potential (TRAP). The uric acid present in saliva correlates with plasma uric acid, suggesting that the former is imported from plasma. There are not statistical differences between uric acid or TRAP values in saliva of smokers and non-smokers. Also, smoking a cigarette does not modify the levels of antioxidants present in saliva.     

Spectrophotometric determination of antioxidant activity

Summary

The spectrophotometric technique for total antioxidant activity (TAA)^1,2 measures the relative abilities of antioxidants to scavenge the 2,2′-azino-bis(3-ethylbenz-thiazoline-6-sulfonic acid) (ABTS) radical cation (ABTS^?+) in comparison with the antioxidant potency of standard amounts of Trolox, the water-soluble vitamin E analogue. This method is based on the progressive consumption of antioxidant activity by ABTS^?+ as it is generated in the reaction cuvette and can be automated with a spectrophotometric analyzer. Several different analytical strategies are possible using the same reagents, enabling the assay system to be used to determine the antioxidant activity of plasma, saliva, lipoprotein fractions, foods and beverages. To determine the activity of pure antioxidant substances, a hydrogen peroxide concentration of 75 μM is used, together with a 6 min measuring time. For biological samples with endogenous peroxidase activity the hydrogen peroxide concentration is increased fivefold and the measuring time shortened to 3.25 min. Assays with improved sensitivity are described for low-density lipoprotein (LDL) preparations and saliva. Use of a spectrophotometric endpoint makes the assay simple to carry out without special laboratory equipment. Measurement at 734 nm avoids a range of potential interfering factors, such as sample turbidity and non-specific absorbance by sample constituents. Current applications of the ABTS antioxidant assay are described and discussed.     

Uric acid concentration in saliva and its changes with the patients receiving treatment for hyperuricemia

To date, few studies have examined uric acid in saliva or dental calculus. The purpose of this study is to examine the uric acid concentration in saliva and serum. Saliva and blood samples were collected from 244 participants. We divided them into four groups: untreated or treated group in normal or abnormal serum uric acid concentration groups. Within the untreated group, Pearson??s correlation coefficient was used to examine the correlation between salivary and serum uric acid concentrations. We compared uric acid concentrations between saliva and serum, or between untreated and treated groups using the paired or unpaired student??s t-test. In the untreated group, uric acid concentrations in saliva and serum were significantly and positively correlated (r?=?0.503, P?<?0.01). Within the untreated group, those with abnormal serum uric acid concentrations had significantly higher uric acid concentrations in serum and saliva compared to those with normal serum uric acid concentrations (P?<?0.01). Within the untreated group, uric acid concentrations in serum were significantly higher than that in saliva (P?<?0.01). Uric acid concentrations in saliva of the treated group were significantly higher than that of the untreated group (P?<?0.01). Within the treated group, uric acid concentrations in saliva were significantly higher than that of serum, particularly in users of benzbromarone (P?<?0.01). Uric acid concentrations in saliva were lower than that in serum among non-users of benzbromarone. In contrast, uric acid concentrations in saliva of patients taking benzbromarone were higher than that in serum. We surmise that URAT1 may influence uric acid excretion in the salivary gland.     

Free-radical processes and the antioxidant system in the realization of the recovery function of sleep

The somnological status of test subjects was assessed. The content of malonic dialdehyde, total peroxidase activity, and concentration of uric acid, urea, and total protein in the saliva of students with insomnia and age-matched (20- to 23-year-old) healthy students were determined during everyday studies and summer exams. It was found that emotional (examination) stress intensifies free-radical processes and leads to a compensatory increase in the activity of the nonenzymatic antioxidant system in the saliva of students. Insomnia is accompanied by an increased level of free-radical processes and increased total protein content in the saliva of test subjects. The recovery anabolic function of sleep is realized via activation of protein synthesis and involvement of low-molecular-weight nonenzymatic antioxidant compounds (uric acid and urea) in the defensive responses of the body under conditions of a significant increase in the intensity of free-radical processes during examination stress combined with insomnia.     

Reactions of Peroxynitrite with Uric Acid: Formation of Reactive Intermediates,Alkylated Products and Triuret,and In Vivo Production of Triuret Under Conditions of Oxidative Stress

Hyperuricemia is associated with hypertension, metabolic syndrome, preeclampsia, cardio-vascular disease and renal disease, all conditions associated with oxidative stress. We hypothesized that uric acid, a known antioxidant, might become prooxidative following its reaction with oxidants; and, thereby contribute to the pathogenesis of these diseases. Uric acid and 1,3-¹⁵N₂-uric acid were reacted with peroxynitrite in different buffers and in the presence of alcohols, antioxidants and in human plasma. The reaction products were identified using liquid chromatography-mass spectrometry (LC-MS) analyses. The reactions generate reactive intermediates that yielded triuret as their final product. We also found that the antioxidant, ascorbate, could partially prevent this reaction. Whereas triuret was preferentially generated by the reactions in aqueous buffers, when uric acid or 1,3-¹⁵N₂-uric acid was reacted with peroxynitrite in the presence of alcohols, it yielded alkylated alcohols as the final product. By extension, this reaction can alkylate other biomolecules containing OH groups and others containing labile hydrogens. Triuret was also found to be elevated in the urine of subjects with preeclampsia, a pregnancy-specific hypertensive syndrome that is associated with oxidative stress, whereas very little triuret is produced in normal healthy volunteers. We conclude that under conditions of oxidative stress, uric acid can form reactive intermediates, including potential alkylating species, by reacting with peroxynitrite. These reactive intermediates could possibly explain how uric acid contributes to the pathogenesis of diseases such as the metabolic syndrome and hypertension.     

Chemiluminescent in vitro estimation of the inhibitory constants of antioxidants ascorbic and uric acids in Fenton’s reaction in urine

The goal of this research was to measure in vitro the inhibitory constants of the antioxidants ascorbic and uric acid in urine, with lucigenin enhanced chemiluminescence (CL) in Fenton’s system. Maximum CL emission is registered in urine containing H₂O₂ (5·10⁻⁴ M), Fe²⁺ (5·10⁻⁵ M), EDTA (5·10⁻⁵ M), and chemical enhancer lucigenin (10⁻⁴ M) at pH 5.5 and 36°C. Ascorbic acid exhibits up to 4-fold stronger antioxidant effect than uric acid. The constants of antioxidant inhibition in urine were measured at concentrations 10⁻³ and 10⁻⁴ M: for ascorbic acid, 5.92 ± 0.04 and 24.05 ± 1.82 μmol·sec⁻¹; for uric acid, 1.60 ± 0.02 and 21.45 ± 0.97 μmol·sec⁻¹, respectively. Three phases of CL kinetics of urine are well observed: spontaneous CL (0–10 sec), fast flash of CL (10–50 sec), and latent period (50–300 sec). The antioxidant efficiency of ascorbic and uric acids in the final stage of catabolic processes in the body is discussed. Published in Russian in Biokhimiya, 2006, Vol. 71, No. 8, pp. 1062–1065.     

Depletion of urate in human nasal lavage following in vitro ozone exposure

Ozone, a strong oxidant present in summer smog, is thought to primarily react with antioxidant molecules found in the epithelial lining fluid of the respiratory tract. In humans, as much as 40% of inhaled ozone can be removed in the nasal cavity where the major extracellular antioxidant has been identified as uric acid. The present study was undertaken to examine urate/oxidant interactions in human nasal lavage fluid following in vitro exposure to ozone at concentrations relevant to the U.K. Lavage fluid was collected from 8 volunteers using a modified Foley catheter which permits prolonged contact of isotonic saline with the anterior nasal cavity. Nasal lavage samples in multiwell plates were exposed to ozone at concentrations of 50, 100 and 250 ppb. Samples were removed at intervals from 15 to 240 min following exposure and assayed for uric acid depletion. Uric acid concentrations in the nasal lavage were found to fall from 8.52 (time zero) to 3.99 μM, 0.05 and 0.07 μM after 240 min at 50, 100 and 250 ppb ozone respectively. At a non-environmentally relevant ozone concentration of 1000 ppb, uric acid was completely depleted after 60 min. Regression analysis showed a linear correlation between rate of loss of urate and ozone concentration (R² = 0.97). A novel, non-invasive technique is described to investigate antioxidant compromise and its importance in individual subjects. We conclude that uric acid in nasal lavage samples is scavenged by ozone in a dose and time dependant manner.     

Determination of uric acid in human saliva by high-performance liquid chromatography with amperometric electrochemical detection

The aim of the present study is to establish a highly sensitive method for the determination of uric acid (UA) in human saliva. The monitoring of UA levels in less invasive biological samples such as saliva is suggested for the diagnosis and therapy of gout, hyperuricemia, and the Lesch-Nyhan syndrome, and for detecting such conditions as alcohol dependence, obesity, diabetes, high cholesterol, high blood pressure, kidney disease, and heart disease. Reversed-phase high-performance liquid chromatography with electrochemical detection (HPLC-ED) was employed for the determination of UA obtained by solid-phase extraction from saliva. To quantify UA, we compared the ED efficiencies of an amperometric ED (Ampero-ED) with a single electrode and a coulometric ED (Coulo-ED) with a multiple electrode array. The results showed that the detection limits (S/N=3) were 3 nM for Ampero-ED and 6 nM for Coulo-ED, and the linearity of the calibration curves of 60-6000 nM had correlation coefficients exceeding 0.999. In addition, the total analytical time was 10 min. In the sample preparation of UA in saliva, an Oasis MAX solid-phase cartridge was used. The recoveries of UA spiked at 0.6 and 3 microM in saliva were above 95% with a relative standard deviation (RSD) of less than 15%. Therefore, the present method may be used in the routine and diagnostic determination of UA in human saliva.     

不同类型冠心病患者血清Hcy、TBIL、hs-CRP、尿酸的表达及临床意义

目的:探讨不同类型冠心病患者血清同型半胱氨酸(Hcy)、总胆红素(TBIL)、高敏C反应蛋白(hs-CRP)、尿酸的水平及临床意义。方法:选取首都医科大学附属北京地坛医院2015年9月-2018年7月收治的冠心病患者132例为冠心病组,根据临床诊断分为稳定型心绞痛52例(SAP组)、不稳定型心绞痛42例(UAP组)、急性心肌梗死38例(AMI组),另选取50例同时期于我院体检的健康志愿者为对照组,检测各组Hcy、TBIL、hs-CRP、尿酸的水平,采用Pearson相关分析Hcy、TBIL、hs-CRP、尿酸水平之间的相关性,采用Logistic回归分析冠心病的影响因素。结果:冠心病组患者的血清Hcy、hs-CRP、尿酸水平显著高于对照组,TBIL水平显著低于对照组(P0.05)。AMI、UAP组患者的血清Hcy、hs-CRP、尿酸水平显著高于SAP组,TBIL水平显著低于SAP组(P0.05),且AMI组患者的血清Hcy、hs-CRP、尿酸水平显著高于UAP组,TBIL水平显著低于UAP组(P0.05)。经Pearson相关分析显示,Hcy与hs-CRP、尿酸呈正相关,与TBIL呈负相关,hs-CRP与尿酸呈正相关(P0.05),TBIL与hs-CRP、尿酸无明显相关性(P0.05)。经Logistic回归分析显示,Hcy、hs-CRP、尿酸、高血压、糖尿病均是冠心病的独立危险因素(P0.05),TBIL是冠心病的保护因素(P0.05)。结论:冠心病患者血清Hcy、hs-CRP、尿酸水平升高,TBIL水平降低,Hcy、TBIL、hs-CRP、尿酸与患者的病情相关,也是冠心病的影响因素。     

Uric Acid: The Oxidant-Antioxidant Paradox

Uric acid, despite being a major antioxidant in the human plasma, both correlates and predicts development of obesity, hypertension, and cardiovascular disease, conditions associated with oxidative stress. While one explanation for this paradox could be that a rise in uric acid represents an attempted protective response by the host, we review the evidence that uric acid may function either as an antioxidant (primarily in plasma) or pro-oxidant (primarily within the cell). We suggest that it is the pro-oxidative effects of uric acid that occur in cardiovascular disease and may have a contributory role in the pathogenesis of these conditions.     

Physiological and histopathological study on the influence of Ocimum basilicum leaves extract on thioacetamide-induced nephrotoxicity in male rats

Kidney disease is a worldwide public health problem that affects millions of people worldwide. Globally, many risk factors for kidney disease progression have been identified. The global prevalence of acute and chronic forms of kidney disease is rising continuously. Nephrotoxicity is defined as rapid dysfunction of kidney due to toxic influence of medications and chemicals. Nephroprotective agents are material that has potential to minimize the effects of nephrotoxic agents. Plants have been shown to be potential therapeutic agents to protect against nephrotoxicity. The purpose of the present study was to evaluate the nephroprotective effect of basil leaves extract against thioacetamide (TAA) in male rats. Experimental male rats were divided into four groups. Rats of the first group were served as controls. Rats of the second group were exposed to TAA. Rats of the third group were treated with basil leaves extract and TAA. Rats of the fourth group were treated with basil leaves extract. After the end of experimental duration (6 Weeks), rats of the second group showed significantly increases of serum creatinine, blood urea nitrogen and uric acid levels, while the levels of serum superoxide dismutase and glutathione were significantly decreased. Histopathologically, renal sections from rats treated with only TAA showed several alterations in the structure of most renal corpuscles including a degeneration of glomeruli and Bowman's capsules. Treatment with basil leaves extract improved the observed biochemical and histopathological changes induced by TAA intoxication. These new findings indicate that the extract of basil leaves represent protective roles on biochemical and histopathological changes induced by TAA toxicity due to its antioxidant activities.     

Characterization of the differentiated antioxidant profile of human saliva

Saliva is armed with various defense mechanisms, such as the immunological and enzymatic defense systems. In addition, saliva has the ability to protect the mucosa against mechanical insults and to promote its healing via the activity of epidermal growth factor. However, another defense mechanism, the antioxidant system, exists in saliva and seems to be of paramount importance. The most interesting finding of the present study was the demonstration of the existence of much higher concentrations of the various salivary molecular and enzymatic antioxidant parameters in the parotid saliva compared with the submandibular/sublingual saliva. For example, peroxidase, superoxide dismutase, uric acid, and total antioxidant status were higher in resting parotid saliva compared with resting submandibular/sublingual saliva by 2405, 235, 245, and 147%, respectively. Another important finding was the distinction between the salivary antioxidant system and the immunological and enzymatic protective systems, as represented by the salivary concentrations of secretory IgA and lysozyme, respectively. These findings suggest that the profound antioxidant capacity of saliva secreted from parotid glands is related either to the different physiological demands related to eating (parotid predominance), to oral integrity maintenance (submandibular/sublingual predominance), or to the high content of deleterious redox-active transitional metal ions present in parotid saliva. This also may signify that our oral cavity environment is only partially protected against oxidative stress during most of the day and night.     

慢性牙周炎患者唾液中IL-17，miR-146a表达水平与疾病程度的相关性分析

目的:观察和分析慢性牙周炎(CP)患者唾液中白细胞介素-17(IL-17)、微小RNA-146a(miR-146a)表达水平与疾病程度的相关性。方法:选取80例CP患者作为病例组,选取80名健康志愿者作为对照组,对两组研究对象唾液样本中的IL-17、miRNA-146a表达水平进行检测和比较。对病例组患者的菌斑指数(PLI)、牙龈指数(GI)、牙周探诊深度(PD)和附着丧失(AL)等牙周病指标进行检测。结果:病例组患者唾液中IL-17、miR-146a表达水平显著高于对照组,两组之间的差异均有统计学意义(P0.05)。随着疾病程度的加剧,患者唾液中IL-17、miR-146a表达水平也逐渐上升,各组之间的差异均有统计学意义(P0.05)。Spearman等级相关分析结果显示,CP患者唾液中IL-17、miR-146a表达水平与牙周炎严重程度均呈正相关关系(P0.05)。直线相关分析结果显示,CP患者唾液中IL-17、miR-146a表达水平与PLI、GI、PD、AL等牙周炎临床指标均呈正相关关系(P0.05)。结论:CP患者表现为唾液中IL-17、miR-146a表达水平的显著升高,其表达水平与疾病严重程度和临床指标均具有相关性。     

The antioxidant and peroxidase activities of saliva in patients with inflammatory periodontal diseases and possibility of their correction

The antioxidant and peroxidase activities of mixed saliva have been investigated during test loads in control group and patients with caries and combination of caries with inflammatory periodontal diseases. Antioxidant activity of saliva was significantly decreased in the last two groups and so its was corrected by means of xidiphone.     

Global Metabolomic Analysis of Human Saliva and Plasma from Healthy and Diabetic Subjects,with and without Periodontal Disease

Recent studies suggest that periodontal disease and type 2 diabetes mellitus are bi-directionally associated. Identification of a molecular signature for periodontitis using unbiased metabolic profiling could allow identification of biomarkers to assist in the diagnosis and monitoring of both diabetes and periodontal disease. This cross-sectional study identified plasma and salivary metabolic products associated with periodontitis and/or diabetes in order to discover biomarkers that may differentiate or demonstrate an interaction of these diseases. Saliva and plasma samples were analyzed from 161 diabetic and non-diabetic human subjects with a healthy periodontium, gingivitis and periodontitis. Metabolite profiling was performed using Metabolon''s platform technology. A total of 772 metabolites were found in plasma and 475 in saliva. Diabetics had significantly higher levels of glucose and α-hydroxybutyrate, the established markers of diabetes, for all periodontal groups of subjects. Comparison of healthy, gingivitis and periodontitis saliva samples within the non-diabetic group confirmed findings from previous studies that included increased levels of markers of cellular energetic stress, increased purine degradation and glutathione metabolism through increased levels of oxidized glutathione and cysteine-glutathione disulfide, markers of oxidative stress, including increased purine degradation metabolites (e.g. guanosine and inosine), increased amino acid levels suggesting protein degradation, and increased ω-3 (docosapentaenoate) and ω-6 fatty acid (linoleate and arachidonate) signatures. Differences in saliva between diabetic and non-diabetic cohorts showed altered signatures of carbohydrate, lipid and oxidative stress exist in the diabetic samples. Global untargeted metabolic profiling of human saliva in diabetics replicated the metabolite signature of periodontal disease progression in non-diabetic patients and revealed unique metabolic signatures associated with periodontal disease in diabetics. The metabolites identified in this study that discriminated the periodontal groups may be useful for developing diagnostics and therapeutics tailored to the diabetic population.     

Effect of surfactants and inducers on increased uricase production under submerged fermentations by Bacillus cereus

Uricase is a clinical enzyme used for the oxidation of uric acid crystals in gout disease. The present study aimed to increase the suitable surfactant-mediated uricase production on induction by different concentrations of inducers. The efficiency of Bacillus cereus to produce extracellular uricase enzyme was studied in uric acid-containing agar plates. Among the studied inducers, uric acid is the potential inducer for uricase production under submerged fermentations (SMF), which induced 19.41?U/ml uricase in medium containing 2.0?g/L of uric acid, however further increase in the uric acid concentration decreased uricase production, which could be because of substrate inhibition. The physical parameters including agitation speed (rpm) and time duration (h) of uricase production were optimized and found to produce optimum uricase at 150?rpm in 26?h of SMF. Among the studied surfactants, nonionic surfactant, polyvinyl alcohol has shown a remarkable increase in the uricase production of 31.58?U/ml, which is a 61% increase under optimized conditions in SMF. The stability of produced uricase was found at pH 7.5 and temperature 30°C. Also the effects of various metal ions (1?mM) on the uricase activity were studied and observed to be inhibitory in nature in the descending order K⁺?>?Ca²⁺?>?Zn²⁺?>?Fe³⁺?>?Ni²⁺?>?Mg²⁺?>?Mn²⁺?>?Cu²⁺.