DNA sequencing with chain terminating inhibitors

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Methods Of Enzymatic Analysis

A complex antioxidant system has been developed in mammals to relieve oxidative stress. However, excessive reactive species derived from oxygen and nitrogen may still lead to oxidative damage to tissue and organs. Oxidative stress has been considered as a conjoint pathological mechanism, and it contributes to initiation and progression of liver injury. A lot of risk factors, including alcohol, drugs, environmental pollutants and irradiation, may induce oxidative stress in liver, which in turn results in severe liver diseases, such as alcoholic liver disease and non-alcoholic steatohepatitis. Application of antioxidants signifies a rational curative strategy to prevent and cure liver diseases involving oxidative stress. Although conclusions drawn from clinical studies remain uncertain, animal studies have revealed the promising in vivo therapeutic effect of antioxidants on liver diseases. Natural antioxidants contained in edible or medicinal plants often possess strong antioxidant and free radical scavenging abilities as well as anti-inflammatory action, which are also supposed to be the basis of other bioactivities and health benefits. In this review, PubMed was extensively searched for literature research. The keywords for searching oxidative stress were free radicals, reactive oxygen, nitrogen species, anti-oxidative therapy, Chinese medicines, natural products, antioxidants and liver diseases. The literature, including ours, with studies on oxidative stress and anti-oxidative therapy in liver diseases were the focus. Various factors that cause oxidative stress in liver and effects of antioxidants in the prevention and treatment of liver diseases were summarized, questioned, and discussed.

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The Role of Oxidative Stress and Antioxidants in Liver Diseases

In addition to the often-cited advantages of using Arabidopsis thaliana as a model system in plant biological research (1), Arabidopsis has many additional characteristics that make it an attractive experimental organism for studying lea d (Pb) accumulation and tolerance in plants. These include its fortuitous familial relationship to many known metal hyperaccumulators (Brassicaceae), as well as similar Pbaccumulation patterns to most other plants. Using nutrient-agar plates, hydroponic culture, and Pb-contaminated soils as growth media, we found significant variation in Arabidopsis thaliana ecotypes in accumulation and tolerance of Pb. In addition, we have found that Pb accumulation is not obligatorily linked with Pb tolerance, suggesti ng that different genetic factors control these two processes. We also screened ethyl methanesulfonate-mutagenized M2 populations and identified several Pb-accumulating mutants. Current characterization of these mutants indicates that their phenotypes are likely due to alteration of general metal ion uptake or translocation processes since these mutants also accumulate many other metals in shoots. We expect that further characterization of the ecotypes and mutants will shed light on the basic genetic and physiological underpinnings of plant-based Pb remediation. 7. Aromatic nitroreduction of acifluorfen in soils, rhizospheres, and pure cultures of rhizobacteria. Zablotowicz, R. M., Locke, M. A., and Hoagland, R. E. Phytoremediation of soil and water contaminants. Washington, DC : American Chemical Society, 1997. p. 38-53. NAL Call #: QD1.A45-no.664 Abstract: Reduction of nitroaromatic compounds to their corresponding amino derivatives is one of several pathways in the degradation of nitroxenobiotics. Our studies with the nitrodiphenyl ether herbicide acifluorfen showed rapid metabolism to am inoacifluorfen followed by incorporation into unextractable soil components in both soil and rhizosphere suspensions. Aminoacifluorfen was formed more rapidly in rhizospheres compared to soil, which can be attributed to higher microbial populations, espec ially of Gram-negative bacteria. We identified several strains of Pseudomonas fluorescens that possess nitroreductase activity capable of converting acifluorfen to aminoacifluorfen. Factors affecting acifluorfen nitroreductase activity in pure cultures an d cell-free extracts, and other catabolic transformations of acifluorfen, ether bond cleavage, are discussed. Plant rhizospheres should be conducive for aromatic nitroreduction. Nitroreduction by rhizobacteria is an important catabolic pathway for the ini tial degradation of various nitroherbicides and other nitroaromatic compounds in soils under Reduction of nitroaromatic compounds to their corresponding amino derivatives is one of several pathways in the degradation of nitroxenobiotics. Our studies with the nitrodiphenyl ether herbicide acifluorfen showed rapid metabolism to am inoacifluorfen followed by incorporation into unextractable soil components in both soil and rhizosphere suspensions. Aminoacifluorfen was formed more rapidly in rhizospheres compared to soil, which can be attributed to higher microbial populations, espec ially of Gram-negative bacteria. We identified several strains of Pseudomonas fluorescens that possess nitroreductase activity capable of converting acifluorfen to aminoacifluorfen. Factors affecting acifluorfen nitroreductase activity in pure cultures an d cell-free extracts, and other catabolic transformations of acifluorfen, ether bond cleavage, are discussed. Plant rhizospheres should be conducive for aromatic nitroreduction. Nitroreduction by rhizobacteria is an important catabolic pathway for the ini tial degradation of various nitroherbicides and other nitroaromatic compounds in soils under phytoremediation management. 8. Ascorbate: a biomarker of herbicide stress in wetland plants. Lytle, T. F. and Lytle, J. S. Phytoremediation of soil and water contaminants. Washington, DC : American Chemical Society, 1997. p. 106-113. NAL Call #: QD1.A45-no.664 Abstract: In laboratory exposures of wetland plants to low herbicide levels (<0.1 micrograms/mL), some plants showed increased total ascorbic acid suggesting a stimulatory effect on ascorbic acid synthesis occurred; at higher herbicide conce ntrations (greater than or equal to 0.1 micrograms/mL) a notable decline in total ascorbic acid and increase in the oxidized form, dehydroascorbic acid occurred. Vigna luteola and Sesbania vesicaria were exposed for 7 and 21 days respectively to atrazine (0.05 to 1 microgram/mL); Spartina alterniflora 28 days at 0.1 micrograms/mL trifluralin; Hibiscus moscheutos 14 days at 0.1 and 1 microgram/mL metolachlor in fresh and brackish water. The greatest increase following low dosage occurred with S. alterniflo ra, increasing from <600 micrograms/g wet wt. total ascorbic acid to >1000 micrograms/g. Ascorbic acid may be a promising biomarker of estuarine plants exposed to herbicide runoff; stimulation of ascorbic acid synthesis may enable some wetland plant s used in phytoremediation to cope with low levels of these compounds. In laboratory exposures of wetland plants to low herbicide levels (<0.1 micrograms/mL), some plants showed increased total ascorbic acid suggesting a stimulatory effect on ascorbic acid synthesis occurred; at higher herbicide conce ntrations (greater than or equal to 0.1 micrograms/mL) a notable decline in total ascorbic acid and increase in the oxidized form, dehydroascorbic acid occurred. Vigna luteola and Sesbania vesicaria were exposed for 7 and 21 days respectively to atrazine (0.05 to 1 microgram/mL); Spartina alterniflora 28 days at 0.1 micrograms/mL trifluralin; Hibiscus moscheutos 14 days at 0.1 and 1 microgram/mL metolachlor in fresh and brackish water. The greatest increase following low dosage occurred with S. alterniflo ra, increasing from <600 micrograms/g wet wt. total ascorbic acid to >1000 micrograms/g. Ascorbic acid may be a promising biomarker of estuarine plants exposed to herbicide runoff; stimulation of ascorbic acid synthesis may enable some wetland plant s used in phytoremediation to cope with low levels of these compounds. 9. Atmospheric nitrogenous compounds and ozone--is NO(x) fixation by plants a possible solution. Wellburn, A. R. New phytol. 139: 1 pp. 5-9. (May 1998). NAL Call #: 450-N42 Descriptors: ozoneair-pollution nitrogen-dioxide nitric-oxide air-quality tolerancebioremediationacclimatizationnutrient-sources nutrient-uptake plantscultivarsgenetic-variation literature-reviews 10. Atrazine degradation in pesticide-contaminated soils: phytoremediation potential. Kruger, E. L., Anhalt, J. C., Sorenson, D., Nelson, B., Chouhy, A. L., Anderson, T. A., and Coats, J. R. Phytoremediation of soil and water contaminants. Washington, DC : American Chemical Society, 1997. p. 54-64. NAL Call #: QD1.A45-no. 664 Abstract: Studies were conducted in the laboratory to determine the fate of atrazine in pesticide-contaminated soils from agrochemical dealer sites. No significant differences in atrazine concentrations occurred in soils treated with atrazine i ndividually or combinations with metolachlor and trifluralin. In a screening study carried out in soils from four agrochemical dealer sites, rapid mineralization of atrazine occurred in three out of eight soils tested, with the greatest amount occurring i n Bravo rhizosphere soil (35% of the applied atrazine after 9 weeks). Suppression of atrazine mineralization in the Bravo rhizosphere soil did not occur with the addition of high concentrations of herbicide mixtures, but instead was increased. Plants had a positive impact on dissipation of aged Studies were conducted in the laboratory to determine the fate of atrazine in pesticide-contaminated soils from agrochemical dealer sites. No significant differences in atrazine concentrations occurred in soils treated with atrazine i ndividually or combinations with metolachlor and trifluralin. In a screening study carried out in soils from four agrochemical dealer sites, rapid mineralization of atrazine occurred in three out of eight soils tested, with the greatest amount occurring i n Bravo rhizosphere soil (35% of the applied atrazine after 9 weeks). Suppression of atrazine mineralization in the Bravo rhizosphere soil did not occur with the addition of high concentrations of herbicide mixtures, but instead was increased. Plants had a positive impact on dissipation of aged atrazine in soil, with significantly less atrazine extractable from Kochia-vegetated soils than from nonvegetated soils. 11. Bacterial inoculants of forage grasses that enhance degradation of 2-chlorobenzoic acid in soil. Siciliano, S. D. and Germida, J. J. Environ toxicol chem. 16: 6 pp. 1098-1104. (June 1997). NAL Call #: QH545.A1E58 Descriptors: polluted-soils bioremediationAbstract: Biological remediation of contaminated soil is an effective method of reducing risk to human and ecosystem health. Bacteria and plants might be used to enhance remediation of soil pollutants in situ. This study assessed the potential of bacteria (12 isolates), plants (16 forage grasses), and plant-bacteria associations (selected pairings) to remediate 2-chlorobenzoic acid (2CBA)-contaminated soil. Initially, grass viability was assessed in 2CBA-contaminated soil. Soil was contaminated wi th 2CBA, forage grasses were grown under growth chamber conditions for 42 or 60 d, and the 2CBA concentration in soil was determined by gas chromatography. Only five of 16 forage grasses grew in 2CBA-treated (816 mg/kg) soil. Growth of Bromus inermis had no effect on 2CBA concentration, whereas Agropyron intermedium, B. biebersteinii, A. riparum, and Elymus dauricus decreased 2CBA relative to nonplanted control soil by 32 to 42%. The 12 bacteria isolates were screened for their ability to promote the germ ination of the five grasses in 2CBA-contaminated soil. Inoculation of A. riparum with Pseudomonas aeruginos

Phytoremediation : 植物による環境/土壌浄化

Mitochondria are the primary intracellular site of oxygen consumption and the major source of reactive oxygen species (ROS), most of them originating from the mitochondrial respiratory chain Among the arsenal of antioxidants and detoxifying enzymes existing in mitochondria, mitochondrial glutathione (mGSH) emerges as the main line of defense for the maintenance of the appropriate mitochondrial redox environment to avoid or repair oxidative modifications leading to mitochondrial dysfunction and cell death mGSH importance is based not only on its abundance, but also on its versatility to counteract hydrogen peroxide, lipid hydroperoxides, or xenobiotics, mainly as a cofactor of enzymes such as glutathione peroxidase or glutathione-S-transferase (GST) Many death-inducing stimuli interact with mitochondria, causing oxidative stress; in addition, numerous pathologies are characterized by a consistent decrease in mGSH levels, which may sensitize to additional insults From the evaluation of mGSH inf

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Mitochondrial glutathione, a key survival antioxidant.

Ameliorating effect of chicory (Chichorium intybus L.) fruit extract against 4-tert-octylphenol induced liver injury and oxidative stress in male rats.

Phytochemical, antioxidant and mineral composition of hydroalcoholic extract of chicory (Cichorium intybus L.) leaves.

The modulatory effect of deltamethrin on antioxidants in mice.

Attenuation of tamoxifen-induced hepatotoxicity by taurine in mice

The modulatory effect of cadmium pretreatment (0.2 mg/kg b.w. i.p.) on deltamethrin-induced oxidative stress and alterations of antioxidants was studied in freshwater fish Channa punctata Bloch. Lipid peroxidation (LPO) was measured as one of the indicators of oxidative stress. Activities of glutathione peroxidase, glutathione reductase, and glutathione-S-transferase were also studied in liver, kidney, and gills. Levels of reduced glutathione (GSH) were measured in various tissues. Fish pretreated with cadmium and subsequently exposed to deltamethrin showed significantly reduced LPO values when compared with deltamethrin-exposed fish. Conversely, in the kidney, a potentiation response was observed. Deltamethrin exposure in fish resulted in significant alteration in activities of glutathione-dependent antioxidant enzymes. In the cadmium preexposed fish, which were subsequently exposed to deltamethrin, antioxidant enzymes showed a tendency toward normalization over deltamethrin-only exposed fish in liver and gills. GSH also showed a similar pattern in liver and gills but in kidney it remained elevated. Cadmium alone had no significant effect on various parameters at the concentration used in this study. When metallothionein (MT) induction was studied, only liver showed an MT-like protein band in sodium dodecylsulfate-polyacrylamide gel electrophoresis analysis. These results demonstrate a modulatory role of cadmium on the oxidative stress and other related parameters in liver and gills. These findings are important in the context of exposure to a mixture of pollutants in aquatic environment.

Hasibur Rehman

Papers

Ameliorating effect of chicory (Chichorium intybus L.) fruit extract against 4-tert-octylphenol induced liver injury and oxidative stress in male rats.

Phytochemical, antioxidant and mineral composition of hydroalcoholic extract of chicory (Cichorium intybus L.) leaves.

The modulatory effect of deltamethrin on antioxidants in mice.

Attenuation of tamoxifen-induced hepatotoxicity by taurine in mice

Modulatory Effect of Cadmium Exposure on Deltamethrin-Induced Oxidative Stress in Channa punctata Bloch