• Asian Pacific Journal of Cancer Prevention
• /
• v.16 no.16
• /
• pp.6967-6972
• /
• 2015

Background: Cigarette smoking and tobacco chewing are common modes of consuming tobacco all over the world. Parents need to be aware that germ cell integrity is vital for birth of healthy offspring as biological parenting begins much before birth of a child and even before conception. The present study was conducted to determine the etiology of non-familial sporadic heritable retinoblastoma (NFSHRb), by evaluating oxidative sperm DNA damage in fathers due to use of tobacco (smoking and chewing). Materials and Methods: We recruited 145 fathers of NFSHRb children and 53 fathers of healthy children (controls) in the study. Tobacco history was obtained by personal interview. Seminal reactive oxygen species (ROS) in semen, sperm DNA fragmentation index (DFI) and 8 hydroxy 2' deoxyguanosine (8-OHdG) levels in sperm were evaluated. The RB1 gene was screened in genomic blood DNA of parents of children with NFSHRb and controls. Odds ratios (ORs) derived from conditional logistic regression models. Results: There was significant difference in the levels of ROS (p<0.05), DFI (p<0.05) and 8-OHdG (p<0.05) between tobacco users and non-users. The OR of NFSHRb for smokers was 7.29 (95%CI 2.9-34.5, p<0.01), for tobacco chewers 4.75 (2.07-10.9, p<0.05) and for both 9.11 (3.79-39.2; p<0.01). Conclusions: This study emphasizes the adverse effect of tobacco on the paternal genome and how accumulation of oxidative damage in sperm DNA may contribute to the etiology of NFSHRb. In an ongoing parallel study in our laboratory, 11 of fathers who smoked underwent. Meditation and yoga interventions, showed significant decline in levels of highly mutagenic oxidised DNA adducts after 6 months. Thus our lifestyle and social habits impact sperm DNA integrity and simple interventions like yoga and meditation are therapeutic for oxidative damage to sperm DNA.

• Animal Bioscience
• /
• v.36 no.7
• /
• pp.1022-1033
• /
• 2023

Objective: p66Shc, a 66 kDa protein isoform encoded by the proto-oncogene SHC, is an essential intracellular redox homeostasis regulatory enzyme that is involved in the regulation of cellular oxidative stress, apoptosis induction and the occurrence of multiple age-related diseases. This study investigated the expression profile and functional characteristics of p66Shc during preimplantation embryo development in sheep. Methods: The expression pattern of p66Shc during preimplantation embryo development in sheep at the mRNA and protein levels were studied by quantitative real-time polymerase chain reaction (RT-qPCR) and immunofluorescence staining. The effect of p66Shc knockdown on the developmental potential were evaluated by cleavage rate, morula rate and blastocyst rate. The effect of p66Shc deficiency on reactive oxygen species (ROS) production, DNA oxidative damage and the expression of antioxidant enzymes (e.g., catalase and manganese superoxide dismutase [MnSOD]) were also investigated by immunofluorescence staining. Results: Our results showed that p66Shc mRNA and protein were expressed in all stages of sheep early embryos and that p66Shc mRNA was significantly downregulated in the 4-to 8-cell stage (p<0.05) and significantly upregulated in the morula and blastocyst stages after embryonic genome activation (EGA) (p<0.05). Immunofluorescence staining showed that the p66Shc protein was mainly located in the peripheral region of the blastomere cytoplasm at different stages of preimplantation embryonic development. Notably, serine (Ser36)-phosphorylated p66Shc localized only in the cytoplasm during the 2- to 8-cell stage prior to EGA, while phosphorylated (Ser36) p66Shc localized not only in the cytoplasm but also predominantly in the nucleus after EGA. RNAi-mediated silencing of p66Shc via microinjection of p66Shc siRNA into sheep zygotes resulted in significant decreases in p66Shc mRNA and protein levels (p<0.05). Knockdown of p66Shc resulted in significant declines in the levels of intracellular ROS (p<0.05) and the DNA damage marker 8-hydroxy2'-deoxyguanosine (p<0.05), markedly increased MnSOD levels (p<0.05) and resulted in a tendency to develop to the morula stage. Conclusion: These results indicate that p66Shc is involved in the metabolic regulation of ROS production and DNA oxidative damage during sheep early embryonic development.

• Journal of Nutrition and Health
• /
• v.35 no.3
• /
• pp.279-290
• /
• 2002

This study was performed to assess age-related changes in DNA damage and antioxidative capacity in 4, 8, 12, 16, 20, and 24 months old Sprague-Dawley male rats. The following were measured the degree of oxidative DNA damage as indicated by levels of 8-hydroxy-2'-deoxyguanosine (80HdG) in the kidney ; the peroxidized lipid concentrations in the plasma and the liver, as indicated by the levels of thiobarbituric acid reactive substances (TBARS); and the levels of antioxidant enzyme activities in the erythrocytes and the liver. Both body weight (BW) and epididymal fat pad (EFP) weight per BW increased with age until 16 months, then decreased slightly from 20 to 24 months. However, the weights of the liver, kidney and spleen per BW decreased with age. Concentrations of 8-OHdG in the kidney increased with age, only slightly front 4 to 16 months, and then markedly from 16 to 24 months. TBARS concentrations in the plasma and liver were shown to increase with age, being lowest in the 4 month-old group and highest in the 24 month-old group. Superoxide dismutase (SOD) activity in the erythrocytes increased with age Catalase activity in the erythrocytes increased from 4 to 16 months, then decreased from 20 to 24 months. Glutathione peroxidase (GSH-Px) activity in the erythrocytes showed no age-related change. Liver SOD activity decreased with age, particularly from 16 to 20 months, but catalase and GSH-Px activities in the liver showed no significant changes. These results showed that during the normal aging of SD rats, DNA damage in the kidney and TBARS concentrations in the plasma and liver increased with age, particularly after 16 months, and the imbalance of antioxidative enzyme activities in the erythrocytes accelerated with age.

• The Korea Journal of Herbology
• /
• v.31 no.1
• /
• pp.7-15
• /
• 2016

Objectives : Polygalae Radix (POL) has an ameliorating effect on learning and memory impairment caused by cerebral hypoperfusion. In regard to POL's action mechanism, this study was carried out to investigate the effects of POL on oxidative damage and neuronal apoptosis induced by cerebral hypoperfusion in rats.Methods : The cerebral hypoperfusion was induced by permanent bilateral common carotid artery occlusion (pBCAO) in Sprague-Dawley rats. POL was administered orally once a day (130 mg/kg of water-extract) for 28 days starting at 4 weeks after the pBCAO. Superoxide dismutase (SOD) activities and malondialdehyde (MDA) levels in the brain tissue were measured using ELISA method. Expressions of 4-hydroxynonenal (4HNE) and 8-hydroxy-2'- deoxyguanosine (8-OHdG) were observed using immunohistochemistry. In addition, neuronal apoptosis was evaluated with Cresyl violet staining, TUNEL labeling, and immunohistochemistry against Bax and caspase-3.Results : POL treatment significantly increased SOD activities and significantly reduced MDA levels in the cerebral cortex. The up-regulations of 4HNE and 8-OHdG expression caused by pBCAO in the CA1 of hippocampus were significantly attenuated by POL treatment. POL treatment also restored the reduction of CA1 thickness and CA1 neurons caused by pBCAO and significantly attenuated the apoptotic markers including TUNEL-positive cells, Bax, and caspase-3 expression in the CA1 of hippocampus.Conclusions : The results show that POL attenuated the oxidative damage in brain tissue and neuronal apoptosis in the hippocampus caused by the cerebral hypoperfusion. It suggests that POL can be a beneficial medicinal herb to treat the brain diseases related to cerebral hypoperfusion.

• Journal of Nutrition and Health
• /
• v.37 no.8
• /
• pp.633-644
• /
• 2004

This study was performed to investigate effect of dried powders and ethanol extracts of garlic flesh and peel on antioxidative capacity in 16-month- old rats. Forty Sprague-Dawley male rats weighing 618.1$\pm$6.5g were blocked into five groups according to body weight and raised for 3 months with experimental diets containing 5% (w/w) of dried powders of garlic flesh or peel, or ethanol extracts from equal amount of each dried powder. Total polyphenols, flavonoids, /3 -carotene, vitamin C, vitamin E, and total antioxidant status (TAS) levels were determined in garlic preparations. Thiobarbituric acid reactive substances (TBARS) levels in plasma, liver and VLDL + LDL fraction, oxidative DNA damage (8-hydroxy-2' -deoxyguanosine, 80HdG) in kidney, xanthine oxidase (XO) activities in plasma and liver, superoxide dismutase (SOD) activities in erythrocyte and liver, and carotenoid concentration, and total antioxidant status (TAS) in plasma were measured. Total polyphenols and flavonoids contents in garlic preparations were highest in peel ethanol extract. Vitamin C content was not different significantly among preparations, but peel powder contains slightly more vitamin C. The content of $\beta$-carotene was highest in peel ethanol extract and vitmain E content was highest in flesh ethanol extract. The highest level of TAS was observed in peel ethanol extract. Plasma TBARS levels in all the experimental groups were found to be significantly lower than control group, and TBARS concentration in VLDL + LDL fraction was decreased in all the experimental groups in comparison to control group. Also levels of 80HdG in kidney in experimental groups were lower than that of control group. Plasma and liver XO activities were. decreased in all experimental groups, and erythrocyte and liver SOD activities were higher in experimental groups compared to control group. All experimental groups also showed higher plasma TAS levels than control group. Especially, garlic flesh powder group was significantly lower in plasma and liver XO activities, and significantly higher in erythrocyte and liver SOD activities than control group. Moreover, plasma TBARS level and kidney 8OHdG level were decreased in flesh powder group. In conclusion, garlic diets showed effect of improving antioxidative capacity in 16-month old rats, especially, garlic flesh powder was prominent in inhibiting XO activity, promoting SOD activity and decreasing kidney 8OHdG level among experimental groups.

• Nutrition Research and Practice
• /
• v.9 no.2
• /
• pp.165-173
• /
• 2015

BACKGROUND/OBJECTIVES: This study addressed the question whether the composition of supposedly 'healthy' or 'unhealthy' dietary regimes has a calorie-independent short-term effect on biomarkers of metabolic stress and vascular risk in healthy individuals. SUBJECTS/METHODS: Healthy male volunteers (age $29.5{\pm}5.9years$, n = 39) were given a standardized baseline diet for two weeks before randomization into three groups of different dietary regimes: fast food, Mediterranean and German cooking style. Importantly, the amount of calories consumed per day was identical in all three groups. Blood samples were analyzed for biomarkers of cardiovascular risk and metabolic stress after two weeks of the baseline diet and after two weeks of the assigned dietary regime. RESULTS: No dietary intervention affected the metabolic or cardiovascular risk profile when compared in-between groups or compared to baseline. Subjects applied to the Mediterranean diet showed a statistically significant increase of uric acid compared to baseline and compared to the German diet group. Plasma concentrations of urea were significantly higher in both the fast food group and the Mediterranean group, when compared to baseline and compared to the German diet group. No significant differences were detected for the levels of vitamins, trace elements or metabolic stress markers (8-hydroxy-2-deoxyguanosine, malondialdehyde and methylglyoxal, a potent glycating agent). Established parameters of vascular risk (e.g. LDL-cholesterol, lipoprotein(a), homocysteine) were not significantly changed in-between groups or compared to baseline during the intervention period. CONCLUSIONS: The calorie-controlled dietary intervention caused neither protective nor harmful short-term effects regarding established biomarkers of vascular or metabolic risk. When avoiding the noxious effects of overfeeding, healthy individuals can possess the metabolic capacity to compensate for a potentially disadvantageous composition of a certain diet.