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Gestational Vitamin B12 Supplementation do not Cause Genomic Instability in Rat Dams and Their Offspring
Nutrition & Health 182 Effect of Piquiá (Caryocar Villosum) Pulp Fruit on Oxidative Stress, Ephx2 and Tp53 Gene Expressions in Liver of Rats 1
Mara Ribeiro Almeida , Alexandre Ferro Aissa1, Joana Darc Castania Darim1, Tarsila D.U.H Gomes1, Renan Campos Chisté1, Adriana Zerlotti Mercadante1, Lusânia Maria Greggi Antunes1, and Maria Lourdes Pires Bianchi1 1 Ribeirão Preto, Brazil A diet rich in fruits has an important role in health promotion and has been related to disease prevention by different mechanisms. The antioxidants compounds present in fruits are indicated as the major responsible for these beneficial effects. Piquiá (Caryocar villosum) is native from Amazon forests and their fruit pulp is rich in carotenoids, phenols and flavonoids compounds. Previous study demonstrated that piquiá pulp fruit has protective effect against DNA damage induced by doxorubicin (DXR). The aim of this study was to investigate whether the piquiá treatment has influence in oxidative stress and also whether this fruit modulates tumor protein p53 (Tp53) and epoxide hydrolase 2 (Ephx2) gene expressions. Ephx2 participates in the arachidonic acid metabolism and consequently is indirectly involved in oxidative stress and Tp53 has an important role in genomic integrity and is also involved in generation of reactive oxygen species. Wistar rats received piquiá pulp fruit (75 or 300 mg/kg) by gavage for 14 days, followed by injection of saline or doxorubicin (DXR) (15 mg/kg b.w.,i.p.), just after the last gavage and 24 hours before euthanasia. The animals were divided into six treatment groups: negative control (saline solution); positive control (DXR); piquiá pulp treatments and piquiá + DXR groups. Oxidative stress was evaluated by thiobarbituric acid-reactive substances (TBARS) and reduced glutathione (GSH) and the gene expression by real time PCR. All assays were performed in liver. Piquiá increased the GSH levels but did not change the TBARS content. The piquiá + DXR groups exhibited no difference in these biochemical parameters when compared to DXR group. Piquiá did not change the Ephx2 and Tp53 gene expressions and also abolished the DXR-induced increased in Ephx2 mRNA levels. The highest dose of piquiá in the piquiá + DXR group decreased the Tp53 gene expression compared to DXR group. These results demonstrate that piquiá pulp has influence in GHS levels and also has effect at genomic levels modulating the Ephx2 and Tp53 gene expressions. Financial support: FAPESP and CAPES.
doi:10.1016/j.freeradbiomed.2012.10.330
183
doi:10.1016/j.freeradbiomed.2012.10.331
184 Gestational Vitamin B12 Supplementation do not Cause Genomic Instability in Rat Dams and Their Offspring
Paula Lumy Takeuchi1, Mara Ribeiro Almeida1, Regislaine Valéria Burim1, Graciela Cristina Santos1, Joana Darc Castania Darin1, 1 Lusânia Maria Greggi Antunes1, and Maria de Lourdes Pires Bianchi 1 Ribeirão Preto-USP, Brazil Maternal nutrition may affect physiological and metabolic parameters in the fetus and the oxidative stress has been implicated as a key link involved in these events. Oxidative stress also is often related to cause DNA damage. Vitamin B12, which has antioxidant properties, plays important role in DNA metabolism and has been identified as a crucial nutrient for fetal development. Variations in vitamin B12 intake are associated to genome integrity and particularly its deficiency can lead to an elevated DNA damage. This study investigated the effect of maternal vitamin B12 supplementation on genomic instability by micronucleus test in peripheral blood and comet assay in liver and peripheral blood cells, both in dams and their offsprings. All groups received standard commercial rat chow during the experimental period. Dams were divided in two groups, a control group and a supplemented group, which received extra vitamin B12 (2.4 μg/day) by gavage before mating and during gestation and lactation (10 weeks). Offspring were euthanized ten days after birth. Both micronucleus test and comet assay showed no changes among the groups in all tissues evaluated. These results suggest that maternal vitamin B12 supplementation do not increase genomic instability in peripheral blood and liver cells of dams and offspring. Financial support: FAPESP and CNPq.
doi:10.1016/j.freeradbiomed.2012.10.332
A Natural H2S Donor Found in Garlic Attenuates Ischemia-Reperfusion Damage in Neurons by Protecting Mitochondrial Respiration. 1
185
Gloria A. Benavides , Qiuli Liang1, Jianhua Zhang1, and Victor M. Darley-Usmar1 1 University of Alabama at Birmingham New therapies to reduce the cerebral ischemia-reperfusion damage are of interest due to the severity of the long-term consequences of stroke. Hydrogen Sulfide (H2S) has been shown to decrease the levels of reactive oxygen species (ROS) during ischemia-reperfusion conditions but the mechanism of protection remains unknown. In the present study we tested the efficacy of the H2S donor Diallyl Disulfide (DADS), one of the
S82
major organosulfur compounds found in garlic, to decrease the mitochondrial damage caused by hypoxia-reperfusion. Using the Extracellular Flux Analyzer we measured mitochondrial respiratory function in isolated primary cortical neurons in a model of ischemia-reperfusion. DADS at 100 μM did not modify basal mitochondrial respiration under the ischemic phase but decreased damage in the reoxygenation phase compared to control. DetaNONOate (an NO donor; 125 μM) was used to evaluate the role of ROS/RNS in mitochondrial damage under hypoxiareperfusion. NO decreases basal respiration during hypoxia. During the reoxygenation phase, NO impairs mitochondria respiration and this was partially prevented by DADS. On reoxygenation NO exacerbates the inhibition of mitochondrial function and this was also prevented by DADS. We conclude that diallyl disulfide, a natural H2S donor found in garlic, protects mitochondrial function following neuronal ischemia-reperfusion.
Consumption of Cloudy Apple Juice Increases Capacity to Protect DNA Against Damage Induced by Iron Ions in Healthy Humans 1
Karlis Briviba 1 Max Rubner-Institut, Federal Research Institute of Nutrition and Food, Germany Epidemiological and experimental studies show that a high intake of fruits can reduce the risk of cancers. DNA damage plays an important role in the initiation of carcinogenesis. The effect of
SFRBM 2012
Mara Ribeiro Almeida , Alexandre Ferro Aissa1, Joana Darc Castania Darim1, Tarsila D.U.H Gomes1, Renan Campos Chisté1, Adriana Zerlotti Mercadante1, Lusânia Maria Greggi Antunes1, and Maria Lourdes Pires Bianchi1 1 Ribeirão Preto, Brazil A diet rich in fruits has an important role in health promotion and has been related to disease prevention by different mechanisms. The antioxidants compounds present in fruits are indicated as the major responsible for these beneficial effects. Piquiá (Caryocar villosum) is native from Amazon forests and their fruit pulp is rich in carotenoids, phenols and flavonoids compounds. Previous study demonstrated that piquiá pulp fruit has protective effect against DNA damage induced by doxorubicin (DXR). The aim of this study was to investigate whether the piquiá treatment has influence in oxidative stress and also whether this fruit modulates tumor protein p53 (Tp53) and epoxide hydrolase 2 (Ephx2) gene expressions. Ephx2 participates in the arachidonic acid metabolism and consequently is indirectly involved in oxidative stress and Tp53 has an important role in genomic integrity and is also involved in generation of reactive oxygen species. Wistar rats received piquiá pulp fruit (75 or 300 mg/kg) by gavage for 14 days, followed by injection of saline or doxorubicin (DXR) (15 mg/kg b.w.,i.p.), just after the last gavage and 24 hours before euthanasia. The animals were divided into six treatment groups: negative control (saline solution); positive control (DXR); piquiá pulp treatments and piquiá + DXR groups. Oxidative stress was evaluated by thiobarbituric acid-reactive substances (TBARS) and reduced glutathione (GSH) and the gene expression by real time PCR. All assays were performed in liver. Piquiá increased the GSH levels but did not change the TBARS content. The piquiá + DXR groups exhibited no difference in these biochemical parameters when compared to DXR group. Piquiá did not change the Ephx2 and Tp53 gene expressions and also abolished the DXR-induced increased in Ephx2 mRNA levels. The highest dose of piquiá in the piquiá + DXR group decreased the Tp53 gene expression compared to DXR group. These results demonstrate that piquiá pulp has influence in GHS levels and also has effect at genomic levels modulating the Ephx2 and Tp53 gene expressions. Financial support: FAPESP and CAPES.
doi:10.1016/j.freeradbiomed.2012.10.330
183
doi:10.1016/j.freeradbiomed.2012.10.331
184 Gestational Vitamin B12 Supplementation do not Cause Genomic Instability in Rat Dams and Their Offspring
Paula Lumy Takeuchi1, Mara Ribeiro Almeida1, Regislaine Valéria Burim1, Graciela Cristina Santos1, Joana Darc Castania Darin1, 1 Lusânia Maria Greggi Antunes1, and Maria de Lourdes Pires Bianchi 1 Ribeirão Preto-USP, Brazil Maternal nutrition may affect physiological and metabolic parameters in the fetus and the oxidative stress has been implicated as a key link involved in these events. Oxidative stress also is often related to cause DNA damage. Vitamin B12, which has antioxidant properties, plays important role in DNA metabolism and has been identified as a crucial nutrient for fetal development. Variations in vitamin B12 intake are associated to genome integrity and particularly its deficiency can lead to an elevated DNA damage. This study investigated the effect of maternal vitamin B12 supplementation on genomic instability by micronucleus test in peripheral blood and comet assay in liver and peripheral blood cells, both in dams and their offsprings. All groups received standard commercial rat chow during the experimental period. Dams were divided in two groups, a control group and a supplemented group, which received extra vitamin B12 (2.4 μg/day) by gavage before mating and during gestation and lactation (10 weeks). Offspring were euthanized ten days after birth. Both micronucleus test and comet assay showed no changes among the groups in all tissues evaluated. These results suggest that maternal vitamin B12 supplementation do not increase genomic instability in peripheral blood and liver cells of dams and offspring. Financial support: FAPESP and CNPq.
doi:10.1016/j.freeradbiomed.2012.10.332
A Natural H2S Donor Found in Garlic Attenuates Ischemia-Reperfusion Damage in Neurons by Protecting Mitochondrial Respiration. 1
185
Gloria A. Benavides , Qiuli Liang1, Jianhua Zhang1, and Victor M. Darley-Usmar1 1 University of Alabama at Birmingham New therapies to reduce the cerebral ischemia-reperfusion damage are of interest due to the severity of the long-term consequences of stroke. Hydrogen Sulfide (H2S) has been shown to decrease the levels of reactive oxygen species (ROS) during ischemia-reperfusion conditions but the mechanism of protection remains unknown. In the present study we tested the efficacy of the H2S donor Diallyl Disulfide (DADS), one of the
S82
major organosulfur compounds found in garlic, to decrease the mitochondrial damage caused by hypoxia-reperfusion. Using the Extracellular Flux Analyzer we measured mitochondrial respiratory function in isolated primary cortical neurons in a model of ischemia-reperfusion. DADS at 100 μM did not modify basal mitochondrial respiration under the ischemic phase but decreased damage in the reoxygenation phase compared to control. DetaNONOate (an NO donor; 125 μM) was used to evaluate the role of ROS/RNS in mitochondrial damage under hypoxiareperfusion. NO decreases basal respiration during hypoxia. During the reoxygenation phase, NO impairs mitochondria respiration and this was partially prevented by DADS. On reoxygenation NO exacerbates the inhibition of mitochondrial function and this was also prevented by DADS. We conclude that diallyl disulfide, a natural H2S donor found in garlic, protects mitochondrial function following neuronal ischemia-reperfusion.
Consumption of Cloudy Apple Juice Increases Capacity to Protect DNA Against Damage Induced by Iron Ions in Healthy Humans 1
Karlis Briviba 1 Max Rubner-Institut, Federal Research Institute of Nutrition and Food, Germany Epidemiological and experimental studies show that a high intake of fruits can reduce the risk of cancers. DNA damage plays an important role in the initiation of carcinogenesis. The effect of
SFRBM 2012