Oxidative DNA Damage In Lymphocytes Research Articles

Oxidative Stress, Folate Receptor Autoimmunity, and CSF Findings in Severe Infantile Autism.

Background Biomarkers such as oxidative stress, folate receptor alpha (FRα) autoimmunity, and abnormal brain serotonin turnover are common in autism. Methods Oxidative stress biomarkers with pro- and antioxidants were measured in the severe form of infantile autism (n = 38) and controls (n = 24). Children and parents had repeated testing for serum FR autoantibodies, spinal fluid dopamine and serotonin metabolites, pterins, and N5-methyltetrahydrofolate (MTHF). Statistical analysis assessed correlations between variables. Genetic analysis included the SLC6A4 and SLC29A4 genes encoding synaptic serotonin reuptake proteins. Results Compared to controls, the autism group showed a significant increase in oxidative DNA damage in lymphocytes, plasma ceruloplasmin and copper levels with a high copper/zinc ratio, thiol proteins, and superoxide dismutase (SOD) activity. Vitamin C levels were significantly diminished. In most autistic patients, the vitamin A (64%) and D (70%) levels were low. Serum FR autoantibodies fluctuating over 5–7 week periods presented in 68% of all autistic children, 41% of parents vs. 3.3% of control children and their parents. CSF showed lowered serotonin 5-hydroxyindole acetic acid (5HIAA) metabolites in 13 (34%), a low 5HIAA to HVA (dopamine metabolite) ratio in 5 (13%), low 5HIAA and MTHF in 2 (5%), and low MTHF in 8 patients (21%). A known SLC6A4 mutation was identified only in 1 autistic child with low CSF 5HIAA and a novel SLC29A4 mutation was identified in identical twins. Low CSF MTHF levels among only 26% of subjects can be explained by the fluctuating FR antibody titers. Two or more aberrant pro-oxidant and/or antioxidant factors predisposed to low CSF serotonin metabolites. Three autistic children having low CSF 5HIAA and elevated oxidative stress received antioxidative supplements followed by CSF 5HIAA normalisation. Conclusion In autism, we found diverse combinations for FR autoimmunity and/or oxidative stress, both amenable to treatment. Parental and postnatal FR autoantibodies tend to block folate passage to the brain affecting folate-dependent pathways restored by folinic acid treatment, while an abnormal redox status tends to induce reduced serotonin turnover, corrected by antioxidant therapy. Trial Registration. The case-controlled study was approved in 2008 by the IRB at Liège University (Belgian Number: B70720083916). Lay Summary. Children with severe infantile autism frequently have serum folate receptor autoantibodies that block the transport of the essential vitamin folate across the blood-brain barrier to the brain. Parents are often asymptomatic carriers of these serum folate receptor autoantibodies, which in mothers can block folate passage across the placenta to their unborn child. This folate deficiency during the child's intrauterine development may predispose to neural tube defects and autism. Oxidative stress represents a condition with the presence of elevated toxic oxygen derivatives attributed to an imbalance between the formation and protection against these toxic reactive oxygen derivatives. Oxidative stress was found to be present in autistic children where these reactive oxygen derivatives can cause damage to DNA, which changes DNA function and regulation of gene expression. In addition, excessive amounts of these toxic oxygen derivatives are likely to damage the enzyme producing the neuromessenger serotonin in the brain, diminished in about 1/3 of the autistic children. Testing children with autism for oxidative stress and its origin, as well as testing for serum folate receptor autoantibodies, could open new approaches towards more effective treatments.

Etomidate is devoid of genotoxicty and mutagenicity in human lymphocytes and in the Salmonella typhimurium/microsomal activation test

No carcinogenesis or mutagenesis studies have been carried out with etomidate. The current study showed that etomidate has weak cytotoxic potential after 48 h exposure in human lymphocytes and has no hemolytic activity. The weak cytotoxicity seems to be related with redox imbalance of etomidate (40.9 and 81.9 μM) treated lymphocytes. At both etomidate concentrations, a slight decrease of the levels of GSH intracellular content and a significant increase in the amount of carbonylated proteins were observed after 48 h. The contribution of oxidative stress to genetic toxicity was only perceived when the enzyme Fpg was applied in the comet assay. Etomidate (40.9 and 81.9 μM) is a weak generator of oxidative DNA damage in lymphocytes. These damages to DNA probably were repaired, since no DNA strand breaks were detected in the standard alkaline comet assay (in the presence or absence of hepatic S9 microsomal fraction) without Fpg. Also, no micronucleated lymphocytes or carrying chromosomal aberrations were observed. Finally, etomidate (2046.8 and 4093.5 μM) was not mutagenic in the Salmonella/microsome mutagenicity assay, which used four Salmonella typhimurium strains (TA97a, TA98, TA100, and TA102) to detect frameshift and base-substitution mutations. In summary, etomidate is a weak oxidative DNA damaging anesthetic and is devoid of mutagenic properties in eukaryotic and prokaryotic models.

DNA repair gene OGG1 polymorphism and its relation with oxidative DNA damage in patients with Alzheimer’s disease

There is considerable evidence that oxidative DNA damage is increased, DNA repair capacity is decreased in patients with Alzheimer’s disease. Base excision repair is the major pathway in removal of oxidative DNA damage. 8-oxo-deoxyguanosine DNA glycosylase 1 (OGG1) is the enzyme which is involved in the first step of this repair process. Alterations in DNA repair capacity may be related with polymorphisms in DNA repair genes. In order to investigate the effect of OGG1 Ser326Cys polymorphism on oxidative DNA damage level, OGG1 genotyping was performed, basal and oxidative DNA damage in lymphocytes and 8-OHdG level in plasma were examined in patients with Alzheimer’s disease. Basal and oxidative DNA damage and 8-OHdG level were measured by OGG1-modified comet assay and enzyme-linked immunoassay, respectively. OGG1 genotyping was performed by polymerase chain reaction- restriction fragment length polymorphism assay. Basal and oxidative DNA damage and plasma 8-OHdG levels were found to be higher in the Alzheimer’s disease group than those in the control group (P < 0.001). In the Alzheimer’s disease group, the levels of oxidative DNA damage was higher in the patients having OGG1 (Ser326Cys + Cys326Cys) genotype than those in the patients having OGG1 Ser326Ser genotype. It was concluded that oxidative DNA damage is increased in patients with Alzheimer’s disease and OGG1 Ser326Cys polymorphism may be responsible for this increase.

Vitamin D Supplementation Reduces Both Oxidative DNA Damage and Insulin Resistance in the Elderly with Metabolic Disorders

Background: Research evidence indicates that vitamin D deficiency is involved in the pathogenesis of insulin resistance (IR) and associated metabolic disorders including hyperglycemia and dyslipidemia. It also suggested that vitamin D deficiency is associated with elevated levels of oxidative stress and its complications. Therefore, the aim of our study was to determine the effect of vitamin D supplementation on DNA damage and metabolic parameters in vitamin D deficient individuals aged >45 with metabolic disorders. Material and Methods: Of 98 initially screened participants, 92 subjects deficient in vitamin D were included in the study. They were randomly assigned to the following group: with vitamin D supplementation (intervention group, n = 48) and without supplementation (comparative group, n = 44). The patients from both groups were divided into two subgroups according to the presence or absence of type 2 diabetes (T2DM). The intervention group was treated with 2000 International Unit (IU) cholecalciferol/day between October and March for three months. At baseline and after three-month supplementation vitamin D concentration (25-OH)D3 and endogenous and oxidative DNA damage were determined. In addition, fast plasma glucose (FPG), fasting insulin, HbA1c and lipid fraction (total cholesterol (TC), low-density lipoprotein cholesterol (LDL), high-density lipoprotein cholesterol (HDL), triglyceride (TG)), as well as anthropometric measurements (weight, height) were gathered. The following IR-related parameters were calculated Homeostatic Model Assesment – Insulin Resistance (HOMA-IR) and TG/HDL ratio. Results: Three-month vitamin D supplementation increased the mean vitamin D concentration to generally accepted physiological level independently of T2DM presence. Importantly, vitamin D exposure decreased the level of oxidative DNA damage in lymphocytes of patients of intervention group. Among studied metabolic parameters, vitamin D markedly increased HDL level, decreased HOMA-IR, TG/HDL ratio. Furthermore, we found that HbA1c percentage diminished about 0.5% in T2DM patients supplemented with vitamin D. Conclusion: The current study demonstrated that daily 2000I U intake of vitamin D for three months decreased the level of oxidative DNA damage, a marker of oxidative stress, independently on T2DM presence. Furthermore, vitamin D reduced metabolic parameters connected with IR and improved glucose and lipid metabolism. Therefore, our results support the assertion that vitamin D, by reducing oxidative stress and improving of metabolic profile, may decrease IR and related diseases.

Genetic damage in environmentally exposed populations to open-pit coal mining residues: Analysis of buccal micronucleus cytome (BMN-cyt) assay and alkaline, Endo III and FPG high-throughput comet assay

DNA and chromosomal damage in individuals occupationally exposed to coal mining residues have repeatedly been reported in lymphocytes and epithelial cells, suggesting a systemic exposure-response in which generation of oxidative damage may play a major role. Nevertheless, the understanding of this mechanism is still incomplete, particularly in regard to environmental exposures. This study aimed to evaluate DNA damage using the cytome assay (BMN-cyt) in buccal cells and its relation to primary and oxidative DNA damage in lymphocytes, assessed by the high-throughput alkaline and modified (FPG-ENDO III) Comet assay in individuals with environmental exposure to coal mining residues in northern Colombia. Considering metals from coal mining activities as the main source of reactive oxygen species (ROS) generation, the concentrations of inorganic elements in blood samples was also assessed. The analysis revealed that frequencies of BMN-cyt parameters related to DNA damage (micronuclei), cytokinesis (binucleated cells) and cell death (condensed chromatin, karyorrhexis, pyknosis and karyolysis) were significantly higher in individuals that were environmentally exposed to coal compared to the unexposed group. The level of % Tail DNA in the alkaline and the modified Comet assay was 4.0 and 4.3 times higher among exposed individuals than in unexposed controls respectively. Increased MN frequencies in buccal cells were correlated with increased %Tail DNA in alkaline and FPG Comet assay. Additionally, exposed individuals had higher concentrations of Cr, Ni, Mn, and Br in the blood compared to unexposed controls. %Tail DNA in alkaline Comet assay was highly correlated with Al, Mn, and Br concentrations, while %Tail DNA in the FPG Comet assay correlated with Mn levels. These results suggest that oxidative damage, particularly purine oxidation, may play an essential role in DNA damage in individuals exposed to coal residues and that some inorganic elements are related to this effect.

Assessment of Oxidative DNA Damage by Alkaline Comet Assay in Human Essential Hypertension.

The objective of the present study was to investigate the antioxidant status and the extent of oxidative DNA damage in lymphocytes and their relation with essential hypertension (EHT). A total of 100 South Indian subjects aged 30-65 were included for the study. Of these 50 were normotensive controls (group-1) with blood pressure ≥120/80mm Hg, 50 were newly diagnosed (group-2) and were not on any antihypertensive drugs, but had systolic blood pressure ranging between 140 and 160mmHg and diastolic blood pressure 95-100mmHg and 50 newly diagnosed essential hypertensive patients underwent drug therapy for 1year was considered as group-3. Enzymatic and non-enzymatic antioxidants significantly decreased and lymphocyte DNA damage was significantly increased in newly diagnosed hypertensive patients compared with control group. The major decrease in DNA damage and significant improvement in enzymatic and non-enzymatic antioxidants were observed after 1year of antihypertensive therapy in treated group compared with newly diagnosed hypertensive patients. Total antioxidant status and lymphocyte DNA damage showed a strong negative correlation in all the three groups. Essential hypertension associated with oxidative stress which in turn causes genotoxic susceptibility to variety of disease including cancer. In the absence of DNA repair process and DNA checkpoint mechanisms, the genomic integrity is susceptible to extensive damage. In our study, increased oxidative DNA damage and decreased antioxidant levels were frequently observed in the newly diagnosed essential hypertensive patients, suggesting that oxidative stress is important in the pathogenesis of EHT. Therefore, the present study has additional clinical implication. Further investigations with large number of patients along with antioxidant supplement are highly warranted.

Oxidative DNA damage in diabetic and mild gestational hyperglycemic pregnant women.

BackgroundPregnant women with mild gestational hyperglycemia present high risk for hypertension, obesity and hyperglycemia, and appeared to reproduce the model of metabolic syndrome in pregnancy, with hyperinsulinemia and insulin resistance. Our clinical studies showed that mild gestational hyperglycemia or gestational diabetes are related to similar adverse maternal and perinatal outcomes. Hyperglycemia and other factors associated with diabetes generate reactive oxygen species that increase DNA damage levels. The aim of this study was to evaluate oxidative DNA damage in lymphocytes of pregnant women with diabetes or mild gestational hyperglycemia.MethodsThe study included 111 pregnant women distributed into three groups based on oral glucose tolerance test (OGTT) and glycemic profiles (GP), as follows: Normal OGTT and GP (control group); Normal OGTT and abnormal GP (mild gestational hyperglycemia group); Abnormal OGTT and GP (diabetic group). Maternal blood samples (5–10 mL) were collected and processed for determination of oxidative DNA damage by the comet assay, using Fpg and Endo III enzymes. Urine samples were also collected for determination of 8-OHdG concentrations by ELISA.ResultsSubjects in the diabetes group presented increased amount of oxidized purines, while mild gestational hyperglycemia women presented with increased oxidized pyrimidines, compared to the control group.ConclusionGestational, overt diabetes and mild gestational hyperglycemia, were all related to increased oxidative DNA damage. Diabetic pregnant women showed increased level of oxidative DNA damage, perhaps mainly due to hyperglycemia. On the other hand, oxidative DNA damage detected in women with mild gestational hyperglycemia might be associated with repercussions from obesity, hypertension and/or insulin resistance. Interestingly, the type of DNA base affected seemed to be dependent on the glycemic profile or oxidative stress.Electronic supplementary materialThe online version of this article (doi:10.1186/1758-5996-7-1) contains supplementary material, which is available to authorized users.

Investigation of the protective effect of Cinnamomum cassia bark extract against H2O2-induced oxidative DNA damage in human peripheral blood lymphocytes and antioxidant activity

ABSTRACT: Cinnamon, one of the most widely used spices in the world, has been shown to have various biological functions including antidiabetic and antitumor activities. Its antidiabetic and antitumor effects were linked with its strong antioxidant activity. In the present study we aimed to investigate the antioxidant activity and possible protective effect of Cinnamomum cassia bark water extract against H2O2-induced oxidative DNA damage. Viability of lymphocytes was determined by Trypan Blue test. For the evaluation of the antioxidant activity, total phenol and flavonoid contents and 2,2-diphenyl-1-picrylhydrazyl (DPPH) inhibitory activity of the extract were determined. DNA damage was determined by alkaline comet assay in human peripheral blood lymphocytes. Lymphocytes exhibited >86 % survival up till the concentration of 800 μg/ml of the extract. Total phenol and flavonoid contents were calculated as 10.6 g ± 0.001 gallic acid equivalents/ 100 g dry weight and 2.25±0.004 g quercetin equivalents/100 g dry weight of the extract, respectively. The extract concentration providing 50 % inhibition of DPPH was found as 76.68 μg/ml. Cinnamomum cassia bark water extract at ≥100 μg/ml concentrations caused significant protection against H2O2-induced oxidative DNA damage in lymphocytes. Our results support the suggestions that Cinnamomum cassia bark water extract could be beneficial as a prophylactic agent in prevention of oxidative stress-related diseases. KEYWORDS: Cinnamomum cassia; DPPH; total phenol; total flavonoid; alkaline comet assay.

홍국(Monascus purpureus)쌀을 첨가한 고콜레스테롤 식이가 흰쥐의 항산화 활성에 미치는 영향

Red yeast rice (RYR) has been known to exhibit various biological effects, including anti-hyperlipidemia, antioxidant, anti-tumor, and anti-inflammtory activities. Oxidative stress is a main risk factor in the development of cardiovascular disease, such as atherosclerosis. Therefore, the aim of this study was to investigate the possible hypolipidemic and antioxidant effect of RYR on rats fed a high-cholesterol diet supplemented with either 0.2%, 1%, or 5% RYR for 4 weeks. We measured lipid profiles in the plasma and liver, antioxidant enzyme activities in plasma and erythrocyte, gene expression of antioxidant enzymes in the liver, and oxidative DNA damage in lymphocytes. The group supplemented with 0.2% RYR had total cholesterol level in plasma decreased by 24%, while the group supplemented with 5% RYR had high-density cholesterol increased by 20% compared to the control. The antioxidant enzyme activities were also affected by RYR supplementation. Total superoxide dismutase activities in plasma significantly decreased by 11% in the 1% RYR group, while these activities in the liver significantly decreased by 16% and 21% in the 1% and 5% supplemented group compared to the control, respectively. Glutathione peroxidase activities in plasma and erythrocytes increased 13% and 48% in the 1% RYR group, respectively. Catalase (CAT) activity in erythrocytes significantly increased by 49% and 68% in the 1% and 5% RYR group compared to the control, respectively. The gene expression of CAT was up-regulated 7.9 fold compared to the control in the 5% RYR supplemented group. These results suggest that RYR can control hyperlipidemia by improving the lipid profile and modulating oxidative stress.

Amelioration of hydrofluoric acid-induced DNA damage by phytochemicals

Hydrofluoric acid (HF), a solution of hydrogen fluoride in water, is widely used in producing refrigerants, drugs, plastics and aluminums. HF is not a strong acid, but it is a highly corrosive acid, capable of dissolving many materials. In this investigation, HF-induced oxidative DNA damage in lymphocytes was evaluated by comet assay. The ameliorative effect of phytochemicals on the HF-induced oxidative DNA damage was also investigated.

The Inhibitory Effect of Phytochemicals on the Oxidative DNA Damage in Lymphocytes by Chrysotile

We investigated the cytotoxicity and oxidative DNA damage by chrysotile, one of the asbestos, in this investigation. Chrysotile enhanced malondialdehyde (MDA) levels and intracellular reactive oxygen speices generation in human airway epithelial cells. Furthermore, asbestos-induced oxidative DNA damage in lymphocytes was evaluated by single cell gel electrophoresis and quantified as DNA tail moment. Notably, phytochemicals such as curcumin, berberine, and sulforaphane presented inhibitory effect on the asbestos-induced oxidative DNA damage in lymphocytes.

Antioxidative vitamins and oxidative lipid and DNA damage in relation to nutrition

Nutrition is a key environmental factor influencing the incidence of chronic age related free radical induced diseases. The better antioxidant status by a sufficient consumption of food commodities with antioxidant compounds helps to minimize oxidative damage, and thus can delay or prevent pathological changes. In three groups of adult apparently healthy non-smoking population of different nutritional habits (93 persons of general population, non-vegetarians, NV; 77 semi-vegetarians, V-S; 64 lacto-ovo-vegetarians, V-LO) plasma concentrations of vitamins B9, C, E and β-carotene, potential prooxidant homocysteine and products of oxidative damage of lipids (total peroxides, malondialdehyde, oxidized LDL) as well as and oxidative DNA damage in lymphocytes (strand breaks, oxidized purines, oxidized pyrimidines) were measured. Vegetarian (V-S, V-LO) vs NV plasma concentrations of vitamin C, vitamin E, lipid standardized vitamin E, β-carotene and vitamin B9 were significantly higher. Plasma total homocysteine values are significantly higher in both vegetarian groups as a consequence of deficient vitamin B12 concentrations. Hyperhomocysteinemia, a marker of systemic or endothelial oxidative stress may promote the production of hydroxyl radicals which are the initiators of lipid oxidation. In spite of 28.5% and 40.6% incidence of hyperhomocysteinemia in V-S and V-LO, the concentrations of total peroxides, malondialdehyde as well as values of oxidized LDL were significantly reduced as a consequence of a better antioxidant status. Parameters of DNA damage as biomarkers of cancer risk in epidemiological studies are significantly reduced in two groups of vegetarians. The findings suggest that dominant consumption of plant food in vegetarians, higher intake of dietary antioxidants and their higher plasma concentrations in comparison to general population may have protective effects against oxidative damage of molecules and a preventive effect on incidence of chronic age related diseases.

Antioxidative Status, DNA Damage and Lipid Profiles in Korean Young Adults by Glutathione S-Transferase Polymorphisms

Oxidative stress leads to the induction of cellular oxidative damage, which may cause adverse modifications of DNA, proteins, and lipids. The production of reactive species during oxidative stress contributes to the pathogenesis of many diseases. Antioxidant defenses can neutralize reactive oxygen species and protect against oxidative damage. The aim of this study was to assess the antioxidant status and the degree of DNA damage in Korean young adults using glutathione s-transferase (GST) polymorphisms. The GSTM1 and GSTT1 genotypes were characterized in 245 healthy young adults by smoking status, and their oxidative DNA damage in lymphocytes and antioxidant status were assessed by GST genotype. General characteristics were investigated by simple questionnaire. From the blood of the subjects, GST genotypes; degree of DNA damage in lymphocytes; the erythrocyte activities of superoxide dismutase, catalase, and glu- tathione peroxidase; plasma concentrations of total peroxyl radical-trapping potential (TRAP), vitamin C, α- and γ-tocopherol, α- and β-carotene and cryptoxanthin, as well as plasma lipid profiles, conjugated diene (CD), GOT, and GPT were analyzed. Of the 245 subjects studied, 23.2% were GSTM1 wild genotypes and 33.4% were GSTT1 wild gen- otype. No difference in erythrocyte activities of superoxide dismutase, catalase, or glutathione peroxidase, and the plas- ma TRAP level, CD, GOT, and GPT levels were observed between smokers and non-smokers categorized by GSTM1 or GSTT1 genotype. Plasma levels of α- and γ-tocopherol increased significantly in smokers with the GSTT1 wild geno- type (p < 0.05); however, plasma level of α-carotene decreased significantly in non-smokers with the GSTM1 wild geno- type (p < 0.05). DNA damage assessed by the Comet assay was significantly higher in non-smokers with the GSTM1 null genotype; whereas DNA damage was significantly lower in non-smokers with the GSTT1 null genotype. Total cho- lesterol and LDL cholesterol levels were significantly higher in non-smokers with the GSTT1 null genotype than those with the GSTT1 wild genotype (p < 0.05). In conclusion, the GSTM1 null genotype or the GSTT1 wild genotype in non-smokers aggravated their antioxidant status through DNA damage of lymphocytes; however, the GSTT1 wild type in non-smok- ers had normal plasma total cholesterol and LDL-cholesterol levels. This finding confirms that GST polymorphisms could be an important determinant of antioxidant status and plasma lipid profiles in non-smoking young adults. Further study is necessary to clarify the antioxidant status and/or lipid profiles of smokers with the GST polymorphism and to conduct a study with significantly more subjects. (Korean J Nutr 2011; 44(1): 16 ~ 28)

Preventive effects of Korean medicinal herbs on the phenanthrene-induced oxidative DNA damages

Phenanthrene-induced oxidative DNA damage in lymphocyte was evaluated using comet assay and the suppressive effects of Korean medicinal herbs on the DNA damage were examined to screen the medicinal herbs that help to detoxify environmental pollutants. Extracts of Acanthopanax, Benincasae and green tea polyphenols effectively inhibited the phenanthrene-induced oxidative DNA damage in lymphocytes among tested herbs in this study. The present study suggests that Korean medicinal herbs by virtue of their antioxidant potential may be used as effective agents to reduce the oxidative DNA damage by phenanthrene.

Antioxidative and Hypolipidemic Effects of Diosgenin, a Steroidal Saponin of Yam (Dioscorea spp.), on High-Cholesterol Fed Rats

Diosgenin (a steroidal saponin of yam) has long been used as a raw material for the industrial production of steroid drugs, and reported to have a hypocholesterolemic effect by suppressing cholesterol absorption and increasing cholesterol secretion. Oxidative stress has been suggested as a main risk factor in the development of atherosclerosis. The aim of this study is to investigate the possible hypolipidemic and antioxidative effect of diosgenin on rats fed with a high-cholesterol diet supplemented with either 0.1% or 0.5% diosgenin for 6 weeks. We measured the lipid profile in the plasma and liver, lipid peroxidation and antioxidative enzyme activities in the plasma, erythrocyte and gene expression of antioxidative enzymes in the liver, and the oxidative DNA damage in lymphocytes. Diosgenin showed a decrease in the plasma and hepatic total cholesterol levels, but increased the plasma high-density lipoprotein (HDL) cholesterol level. Erythrocyte TBARS and lymphocyte DNA damage measured by the comet assay were decreased in the diosgenin supplemented group. Furthermore, diosgenin feeding enhanced the resistance to lymphocyte DNA damage caused by an oxidant challenge with H(2)O(2). The antioxidative enzyme activities were also affected by diosgenin supplementation. Total superoxide dismutase (SOD) in the plasma and liver, glutathione peroxidase (GSH-Px) in erythrocytes, and catalase (CAT) in erythrocytes and liver were significantly increased in the 0.5% diosgenin group. The expression of antioxidative enzymes was up-regulated by diosgenin, the expression of GSH-Px being the highest in the 0.5% diosgenin group. These results suggest that diosgenin could be a very useful compound to control hypercholesterolemia by both improving the lipid profile and modulating oxidative stress.

Chaga mushroom extract inhibits oxidative DNA damage in lymphocytes of patients with inflammatory bowel disease

Inflammatory Bowel Disease (IBD) is partly caused by oxidative stress from free radicals and reduced antioxidant levels. Using hydrogen peroxide to induce oxidative stress in vitro in peripheral lymphocytes we investigated the induction of DNA damage supplemented with ethanolic extract of Chaga mushroom as a protective antioxidant. Lymphocytes were obtained from 20 IBD patients and 20 healthy volunteers. For treatment, a constant H_{2}O_{2 } dose (50 microg/ml) was used with variable doses of Chaga extract (10-500 microg/ml). DNA damage was evaluated in 50 cells per individual and dose using the Comet assay (making 1000 observations per experimental point ensuring appropriate statistical power). Chaga supplementation resulted in a 54.9% (p < 0.001) reduction of H_{2}O_{2 } induced DNA damage within the patient group and 34.9% (p < 0.001) within the control group. Lymphocytes from Crohn's disease (CD) patients had a greater basic DNA damage than Ulcerative Colitis (UC) patients (p < 0.001). Conclusively, Chaga extract reduces oxidative stress in lymphocytes from IBD patients and also healthy individuals when challenged in vitro. Thus, Chaga extract could be a possible and valuable supplement to inhibit oxidative stress in general.

A half-marathon and a marathon run induce oxidative DNA damage, reduce antioxidant capacity to protect DNA against damage and modify immune function in hobby runners

This study investigated whether a 21.1 km (half-marathon) or a 42.195 km (marathon) run modulates DNA damage, antioxidant capacity in lymphocytes and plasma, and the immune system in healthy hobby runners. Ten and 12 volunteers who completed the Baden-Marathon race in Karlsruhe with a running distance of 21.1 km and 41.195 km, respectively, were assessed 10 days before and immediately after the finish. There was no increase in the levels of endogenous DNA strand breaks immediately after half-marathon or marathon races. A statistically significant increase in the levels of oxidative DNA damage in lymphocytes was found using endonuclease III but not formamidopyrimidine glycolase (Fpg). The resistance of DNA to oxidative damage induced by hydrogen peroxide in isolated lymphocytes was significantly decreased after both races. The levels of plasma antioxidants such as α-tocopherol, β-carotene and lycopene were close to, or higher than, those considered optimal for reducing the risk of cardiovascular diseases and there were no significant changes after the races in antioxidant capacity of LDL (lag-time test) or plasma in ORAC, TEAC or paraoxonase assays. The number and percentage of granulocytes and monocytes able to generate oxidative burst were significantly increased after both races, but the lytic activity of NK cells was significantly increased at the end of the half-marathon; no effect was observed in the marathon runners. Thus, oxidative DNA damage in lymphocytes, decreased the antioxidant capacity to protect lymphocytes against DNA strand breaks and increased the formation of reactive species by phagocytes in well-nourished hobby runners indicating moderate oxidative damage during such high-intensity exercise.

Air pollution and cancer: biomarker studies in human populations †

Large cohort studies in the U.S. and in Europe suggest that air pollution may increase lung cancer risk. Biomarkers can be useful to understand the mechanisms and to characterize high-risk groups. Here we describe biomarkers of exposure, in particular DNA adducts as well as markers of early damage, including mutagenicity, other endpoints of genotoxicity and molecular biomarkers of cancer. Several studies found an association between external measures of exposure to air pollution and increased levels of DNA adducts, with an apparent levelling-off of the dose-response relationship. Also, numerous experimental studies in vitro and in vivo have provided unambiguous evidence for genotoxicity of air pollution. In addition, due to the organic extracts of particulate matter [especially various polycyclic aromatic hydrocarbon (PAH) compounds], particulate air pollution induces oxidative damage to DNA. The experimental work, combined with the data on frequent oxidative DNA damage in lymphocytes in people exposed to urban air pollution, suggests 8-oxo-dG as one of the important promutagenic lesions. Lung cancer develops through a series of progressive pathological changes occurring in the respiratory epithelium. Molecular alterations such as loss of heterozygosity, gene mutations and aberrant gene promoter methylation have emerged as potentially promising molecular biomarkers of lung carcinogenesis. Data from such studies relevant for emissions rich in PAHs are also summarized, although the exposure circumstances are not directly relevant to outdoor air pollution, in order to shed light on potential mechanisms of air pollution-related carcinogenesis.

Does a vegetarian diet influence genomic stability?

The vegetarian lifestyle is supposedly healthy, and differences between vegetarians and non-vegetarians in biomarkers related to diseases such as cancer might be expected. To investigate the possible role of different diets in maintaining genomic stability. The vegetarian group, consisting of 24 volunteers (13 women and 11 men), were matched for age and sex with 24 volunteers (12 women and 12 men) with a traditional dietary habit. Among vegetarians there were 13 lacto-ovo-vegetarians (8 women, 5 men) with average length of vegetarian diet 10.8 years (ranging from 5 to 26) and 11 lacto-vegetarians (5 women, 6 men) with average length of vegetarian diet 8.2 years (ranging from 3 to15). All volunteers were nonsmokers, non-consumers of alcohol and had similar education and patterns of physical activity. Chromosome aberrations, micronuclei and DNA damage (strand breaks, oxidised bases and H(2)O(2)-sensitivity) were examined in peripheral blood lymphocytes of vegetarians and non-vegetarians. Plasma antioxidant status was assessed with the FRAP assay. We did not find any differences in percentage of cells with chromosome aberrations or in the frequency of micronuclei between vegetarians and non-vegetarians or between lacto-ovo and lacto-vegetarians. There was no statistically significant difference in total antioxidant capacity between the groups. The group with traditional dietary habits had significantly higher levels of oxidative DNA damage (strand breaks and oxidised purines, P = 0.005) compared with vegetarians. A significant positive correlation between age and oxidative DNA damage (net FPG-sensitive sites) was found in non-vegetarians, while there was an opposite trend towards a negative association in vegetarians. On the other hand chromosome aberrations correlated with age in vegetarians (r = 0.48, P = 0.017) but not in non-vegetarians. Our results indicate that a vegetarian diet can lead to a slight decrease in oxidative DNA damage in lymphocytes, but other markers of genetic stability are not affected. The lowest level of DNA damage was found in lymphocytes of lactovegetarians, (especially oxidised pyrimidines, P = 0.0017), suggesting that this diet provides some protection against oxidative stress.

Detection of oxidative DNA damage in lymphocytes of patients with Alzheimer's disease

Oxidative damage to DNA may play an important role in both normal ageing and in neurodegenerative diseases. The deleterious consequences of excessive oxidations and the pathophysiological role of reactive oxygen species have been intensively studied in Alzheimer's disease. Although the role of oxidative stress in the aetiology of Alzheimer's disease is still not clear, the detection of an increased damage status in the cells of patients could have important therapeutic implications. The levels of oxidative damage in peripheral lymphocytes of 24 Alzheimer's disease patients and of 21 age-matched controls were determined by comet assay applied to freshly isolated blood samples with oxidative lesion-specific DNA repair endonucleases (endonuclease III for oxidized pyrimidines, formamidopyrimidine glycosylase for oxidized purines). It was demonstrated that Alzheimer's disease is associated with elevated levels of oxidized pyrimidines and purines (p<0.0001) as compared with age-matched control subjects. It was also demonstrated that the comet assay is useful as a biomarker of oxidative DNA damage when used with oxidative lesion-specific enzymes.