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Background: Reactive oxygen species released on stimulation by periodontal pathogens cause oxidation of biomolecules and play significant role in periodontal disease pathogenesis. Aim: This study aimed to evaluate the levels of oxidative by?products malondialdehyde (MDA) and 8?hydroxy deoxyguanosine (8?OHdG) as biomarkers in chronic periodontitis patients compared to healthy as well as before and after nonsurgical periodontal therapy. The correlation between biomarkers and clinical attachment level was also evaluated. Settings and Design: A total of 112 subjects were included in this study. The subjects were divided into two groups (Group I included 56 healthy subjects and Group II constituted 56 chronic periodontitis patients) on the basis of clinical periodontal parameters. Materials and Methods: Group I subjects received no treatment and were evaluated once only for clinical and oxidative stress biomarker parameters. Nonsurgical periodontal therapy was carried out in Group II patients and they were evaluated at baseline and 3 months after therapy. Results: Both salivary and serum levels of MDA and 8?OHdG were found to be increased in chronic periodontitis patients as compared to healthy subjects. After nonsurgical periodontal therapy, the levels of MDA and 8?OHdG significantly reduced. Linear correlation between clinical attachment level and oxidative stress parameters was found to be positive and highly significant. Conclusion: It can be concluded that periodontal therapy is effective in improving the oxidative stress condition.
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Objective To study the changes in protein expression levels of 8 oxide of guanine in the temporal and frontal lobes in SAMP8 (senescence accelerated mice)mice versus SAMR1 (control mice).Methods In the study,we collected 12 SAMP8 mice and 12 SAMR1 mice at different month stage(1m,4m,8m,12m)(n=6,each group).After anesthesia,mice were sacrificed.The temporal and frontal lobe tissues of mice were labeled according to the stereotactic anatomical map of mouse brain and taken up for study.After paraffin sections were made,the expression level of 8 oxide of guanine in brain tissues was detected by immunohistochemical method.Results In the same age group at 1 month in the same temporal lobe,the average optical density of 8-oxodG was slightly higher in SAMP8 mice(0.086)than in SAMR1 mice(0.061,t =1.541,P＞0.05).There were significant differences in the mean optical density of 8-oxodG in temporal lobe in 4,8 and 12 months groups between SAMP8 and SAMR1 group(0.101 vs.0.081,0.147 vs.0.109 and 0.176 vs.0.120,t =2.405,2.612 and 5.019,respectively,P ＜0.05).There was no significant difference in the average optical density levels of 8-oxodG in frontal lobe in 1 and 4 months groups between SAMP8 and SAMR1 mice (0.044 vs.0.030,0.062 vs.0.046,t =1.843 and 1.163,respectively,P ＞ 0.05),while the average optical density levels of 8-oxodG in 8 and 12 months groups(0.090 and 0.138)were higher than those of SAMR1 mice in the same age group(0.049 and 0.063)(t =5.327 and 4.88,P＜0.01).There was no significant difference in the average optical density of 8-oxoG in temporal lobe in 1 month group between SAMP8 and SAMR1 mice(0.099 vs.0.066,t =1.956,P ＞0.05).There were significant differences in the average optical density levels of 8-oxoG in temporal lobe in 4,8 and 12 months groups between SAMP8 and SAMR1 mice(0.119 vs.0.076,0.148 vs.0.094 and 0.173 vs.0.101,t =4.033,3.042 and 5.328,respectively,P＜0.01).There was no significant difference in the average optical density levels of 8-oxoG in frontal lobe in 1 month and 4 months groups between SAMP8 and SAMR1 mice(0.049 vs.0.039,0.058 vs.0.056,t =0.713 and 0.172,respectively,P ＞0.05).The average optical density of 8-oxoG in frontal lobe in 8 months and 12 months groups had significant differences between SAMP8 and SAMR1 mice(0.087 vs.0.058,0.12 vs.0.063,t =2.261 and 4.185,P ＜0.05).In addition,we also found that the average optical density of 8 oxide of guanine in same SAMP8 mice at the same age was higher in temporal lobe than in frontal lobe.Conclusions The protein levels of 8-oxodG and 8-oxoG are increased with age,and the damage caused by nucleic acid oxidation is more severe in temporal lobe than in frontal lobe in SAMP8 mice.
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Objective: To evaluate the antioxidant activity of extracts and fractions from Stachys sieboldii Miq., and to examine its effect on the cellular reactive oxygen species (ROS) and glutathione (GSH) production and genomic DNA oxidation in HT-1080 cells. Methods: The ROS generation induced by H
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Objective: To evaluate the antioxidant activity of extracts and fractions from Stachys sieboldii Miq., and to examine its effect on the cellular reactive oxygen species (ROS) and glutathione (GSH) production and genomic DNA oxidation in HT-1080 cells.Methods: The ROS generation induced by H2O2 was measured by the dichlorofluorescein-diacetate assay. GSH levels were measured using a fluorescent method with mBBr. Genomic DNA oxidative damage was measured with levels of oxidative DNA induced by the reaction of ferritin with H2O2.Results: Then-hexane, 85％ aqueous methanol andn-butanol fractions (0.05 mg/mL concentrations) inhibited H2O2-induced ROS generation by 63％, 35％ and 45％, respectively. GSH levels were significantly increased in both acetone+methylene chloride and methanol extracts (P<0.05). Supplementation of cells withn-hexane significantly increased GSH levels at concentrations of 0.05 mg/mL (P<0.05). Both the acetone+methylene chloride and methanol extracts, as well as all fractions significantly inhibited oxidative DNA damage (P<0.05). Conclusions: These results indicate that cellular oxidation was inhibited by then-hexane fraction and this fraction may contain valuable active compounds.
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Oxidative stress, the imbalance between the production of reactive oxygen species (ROS) and antioxidant activity is a major culprit of male infertility. Peroxiredoxins (PRDXs) are major antioxidant enzymes of mammalian spermatozoa and are thiol oxidized and inactivated by ROS in a dose-dependent manner. Their deficiency and/or inactivation have been associated with men infertility. The aim of this study was to elucidate the impact of oxidative stress, generated by the in vivo tert-butyl hydroperoxide (tert-BHP) treatment on rat epididymal spermatozoa during their maturation process. Adult Sprague-Dawley males were treated with 300 μmoles tert-BHP/kg or saline (control) per day intraperitoneal for 15 days. Lipid peroxidation (2-thibarbituric acid reactive substances assay), total amount and thiol oxidation of PRDXs along with the total amount of superoxide dismutase (SOD), motility and DNA oxidation (8-hydroxy-deoxyguanosine) were determined in epididymal spermatozoa. Total amount of PRDXs and catalase and thiol oxidation of PRDXs were determined in caput and cauda epididymis. While animals were not affected by treatment, their epididymal spermatozoa have decreased motility, increased levels of DNA oxidation and lipid peroxidation along with increased PRDXs (and not SOD) amounts. Moreover, sperm PRDXs were highly thiol oxidized. There was a differential regulation in the expression of PRDX1 and PRDX6 in the epididymis that suggests a segment-specific role for PRDXs. In conclusion, PRDXs are increased in epididymal spermatozoa in an attempt to fight against the oxidative stress generated by tert-BHP in the epididymis. These findings highlight the role of PRDXs in the protection of sperm function and DNA integrity during epididymal maturation.
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In order to prepare antioxidant peptide through hydrolyzing low-value protein resources with bacterial extracellular proteases and to discover novel proteases, crude extracellular protease from Pseudoalteromonas sp. SHK1-2 was obtained through fermentation which was used to hydrolyze collagen extracted from Cirrhinus molitorella skin. Small peptide fraction was isolated from hydrolysate by ultrafiltration and Sephadex LH-20 size exclusion chromatography and showed 1, 1-diphenyl-2-picrylhydrazyl radical scavenging activity (35.6%±7%), oxygen radical absorbance capacity and inhibition of DNA oxidation damage. The molecule weight was 776.2 Da, and amino acid sequence was Thr-Ala-Gly-His-Pro- Gly-Thr-His through liquid chromatography mass spectrum. Our findings suggest that peptide obtained from low-value protein of fish waste by hydrolysis with bacterial protease has antioxidant activity.
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Animals , Amino Acid Sequence , Antioxidants , Chemistry , Chromatography, Gel , Collagen , Chemistry , Cyprinidae , Dextrans , Hydrolysis , Oxidation-Reduction , Peptide Hydrolases , Peptides , Chemistry , Skin , ChemistryABSTRACT
Aim: To investigate the role of iron status in cervical carcinogenesis through its involvement in the Haber-Weis and Fenton reactions serving as a pathway to carcinogenesis and using 8-oxo-7, 8- dihydro-2’-deoxyguanosine (8-oxodG) as a marker of DNA oxidation in a population where iron deficiency is prevalent. Study Design: It is a cross sectional study. Place of Study: The patients were recruited from the colposcopy clinic of the University College Hospital (UCH), Ibadan and Obafemi Awolowo University Teaching Hospitals Complex, Ile-Ife, Nigeria. The laboratory investigations were carried out at the Haematology and Chemical Pathology laboratories of UCH, Ibadan and Oxidative Stress Group, Department of Cancer Studies and Molecular Medicine, University of Leicester, Leicester, UK. Methodology: Forty-five subjects with CIN and 41 with normal Pap smear result (non-CIN) were recruited. A structured questionnaire was administered to collect information on demographic characteristics, dietary, social and medical history. Fasting blood sample were collected to assess for serum iron, total iron binding capacity and transferrin saturation. Urine was also collected to analyze for creatinine and 8-oxodG. Results: The CIN subjects had more babies; > 5 than non-CIN subjects (P=.003). The individuals with > 5 children were 4 times more likely to have CIN [OR 3.79 (95% CI 1.3-10.33), P=.01]. CIN subjects had higher serum iron and transferrin saturation than non-CIN subjects. Though the mean urinary 8-oxodG level similar between the two groups, there was a trend towards higher levels in individuals with high grade CIN. Conclusion: High serum iron level was linked to frequent ingestion of iron supplement and may contribute to progression of CIN with a potential role for urinary 8-oxodG as a useful bio indicator of altered iron homeostasis and associated DNA damage.
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OBJECTIVE: The study aimed to evaluate the feasibility and reproducibility of measuring phospholipase C zeta (PLCzeta) using immunostaining in human sperm and to investigate the relationship between PLCzeta immunoreactivity and DNA fragmentation and oxidation in human sperm. METHODS: Semen samples were obtained from participants (n=44) and processed by the conventional swim-up method. Sperm concentration, motility, normal form by strict morphology, DNA fragmentation index assessed by terminal deoxynucleotidyl transferase dUTP nick end labeling method and immunofluorescent expression for 8-hydroxy-2'-deoxyguanosine (8-OHdG) and PLCzeta were assessed. RESULTS: When duplicate PLCzeta tests were performed on two sperm samples from each of the 44 participants, similar results were obtained (74.1+/-9.4% vs. 75.4+/-9.7%). Two measurements of PLCzeta were found to be highly correlated with each other (r=0.759, P<0.001). Immunoreactivity of PLCzeta was not associated with donor's age, sperm concentration, motility, and the percentage of normal form as well as DNA fragmentation index. However, immunoreactivity of PLCzeta showed a significant negative relationship with 8-OHdG immunoreactivity (r=-0.404, P=0.009). CONCLUSION: Measurement of PLCzeta by immunostaining is feasible and reproducible. Lower expression of PLCzeta in human sperm may be associated with higher sperm DNA oxidation status.
Subject(s)
Humans , DNA , DNA Fragmentation , DNA Nucleotidylexotransferase , Semen , Spermatozoa , Type C PhospholipasesABSTRACT
Objective: To investigate the role of water-soluble extract of Salvia fruticosa (Greek sage) (S. fruticosa) leaves in reducing both intrinsic cellular and H
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<p><b>OBJECTIVE</b>To investigate the role of water-soluble extract of Salvia fruticosa (Greek sage) (S. fruticosa) leaves in reducing both intrinsic cellular and H2O2-induced DNA oxidation in cultured human embryonic kidney 293 cells. S. fruticosa, native to the Eastern-Mediterranean basin, is widely used as a medicinal herb for treatment of various diseases.</p><p><b>METHODS</b>Dried leaves of S. fruticosa were extracted in phosphate buffer saline and purified using both vacuum and high pressure filtrations. Each mL of the preparation contained (7.1±1.0) mg of extract. HEK-293 cells were incubated in one set with S. fruticosa extract in the presence of 0.1 mmol/L H2O2, and in the other set with the addition of the extract alone. The DNA oxidation was measured using fluorescence upon fluorescein isothiocyanate derivatization of 8-oxoguanine moieties. The fluorescence was measured using flow cytometry technique.</p><p><b>RESULTS</b>Cells incubated 3 h with 150 µL extract and exposed to 0.1 mmol/L H2O2 showed lower intensity of fluorescence, and thus lower DNA oxidation. Moreover, cells incubated 3 h with 100 µL of the extract showed lower intensity of fluorescence, and thus lower intrinsic cellular DNA oxidation compared to control (without S. fruticosa).</p><p><b>CONCLUSIONS</b>The results from this study suggest that the water-soluble extract of S. fruticosa leaves protects against both H2O2-induced and intrinsic cellular DNA oxidation in human embryonic kidney 293 cells.</p>
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Objective:To investigate the role of water-soluble extract of Salvia fruticosa (Greek sage) (S. fruticosa) leaves in reducing both intrinsic cellular and H2O2-induced DNA oxidation in cultured human embryonic kidney 293 cells. S. fruticosa, native to the Eastern-Mediterranean basin, is widely used as a medicinal herb for treatment of various diseases. Methods: Dried leaves of S. fruticosa were extracted in phosphate buffer saline and purified using both vacuum and high pressure filtrations. Each mL of the preparation contained (7.1±1.0) mg of extract. HEK-293 cells were incubated in one set with S. fruticosa extract in the presence of 0.1 mmol/L H2O2, and in the other set with the addition of the extract alone. The DNA oxidation was measured using fluorescence upon fluorescein isothiocyanate derivatization of 8-oxoguanine moieties. The fluorescence was measured using flow cytometry technique. Results:Cells incubated 3 h with 150 μL extract and exposed to 0.1 mmol/L H2O2 showed lower intensity of fluorescence, and thus lower DNA oxidation. Moreover, cells incubated 3 h with 100 μL of the extract showed lower intensity of fluorescence, and thus lower intrinsic cellular DNA oxidation compared to control (without S. fruticosa). Conclusions: The results from this study suggest that the water-soluble extract of S. fruticosa leaves protects against both H2O2-induced and intrinsic cellular DNA oxidation in human embryonic kidney 293 cells.