Affinity and Effect of Anticancer Drugs on the Redox Reactivity of Hemoglobin.

Hemoglobin's redox reactivity is affected by anticancer drugs of the antitubulin class. Direct binding of these drugs to hemoglobin, with biomedically relevant affinities, is demonstrated. While this interaction is mostly allosteric, in the case of docetaxel, a direct redox reaction is also observed-correlating well with structural differences between the four compounds. A role for Tyr145 in this reactivity is proposed, in line with previous observations of the importance of this amino acid in the reactivity of Hb toward agents of oxidative stress. A susceptibility of vinorelbin (and to a lower extent of paclitaxel) toward peroxide and peroxidase is shown.

Silicon alleviates salt and drought stress of Glycyrrhiza uralensis seedling by altering antioxidant metabolism and osmotic adjustment.

This study was conducted to determine effect and mechanism of exogenous silicon (Si) on salt and drought tolerance of Glycyrrhiza uralensis seedling by focusing on the pathways of antioxidant defense and osmotic adjustment. Seedling growth, lipid peroxidation, antioxidant metabolism, osmolytes concentration and Si content of G. uralensis seedlings were analyzed under control, salt and drought stress [100 mM NaCl with 0, 10 and 20% of PEG-6000 (Polyethylene glycol-6000)] with or without 1 mM Si. Si addition markedly affected the G. uralensis growth in a combined dose of NaCl and PEG dependent manner. In brief, Si addition improved germination rate, germination index, seedling vitality index and biomass under control and NaCl; Si also increased radicle length under control, NaCl and NaCl-10% PEG, decreased radicle length, seedling vitality index and germination parameters under NaCl-20% PEG. The salt and drought stress-induced-oxidative stress was modulated by Si application. Generally, Si application increased catalase (CAT) activity under control and NaCl-10% PEG, ascorbate peroxidase (APX) activity under all treatments and glutathione (GSH) content under salt combined drought stress as compared with non-Si treatments, which resisted to the increase of superoxide radicals and hydrogen peroxide caused by salt and drought stress and further decreased membrane permeability and malondialdehyde (MDA) concentration. Si application also increased proline concentration under NaCl and NaCl-20% PEG, but decreased it under NaCl-10% PEG, indicating proline play an important role in G. uralensis seedling response to osmotic stress. In conclusion, Si could ameliorate adverse effects of salt and drought stress on G. uralensis likely by reducing oxidative stress and osmotic stress, and the oxidative stress was regulated through enhancing of antioxidants (mainly CAT, APX and GSH) and osmotic stress was regulated by proline.

The effects of cadmium on the biochemical and physiological parameters of Eruca sativa.

In this study, Eruca sativa (Rocket) seedlings were treated with different cadmium (Cd) concentrations (0, 150, 300 and 450 µg ∙ g-1). The effects of Cd on lipid peroxidation, enzymatic (APx, CAT, GPX, SOD) and non-enzymatic antioxidants (total ascorbate, dehydroascorbate, ascorbate, non-protein thiol), fresh and dry masses, water content were determined. Also, Cd content of the leaves and the roots were analysed. The highest cadmium accumulation of leaves was at 450 µg ∙ g-1 Cd treatment and the accumulation was 2.62 times greater than those in the roots. The translocation factor was 3.89 at 300 µg ∙ g-1 Cd treatment. Cd treatments caused decreases of fresh, dry mass and water content of leaves and roots. Malondialdehyde content, which is an index of lipid peroxidation, was increased in proportion with the increase in Cd. While there was not change in the activity of GPX according to control, a decrease in activities of SOD, CAT and APX were observed with the increase of cadmium concentration. Although a significant increase in the amounts of non-protein thiol groups and proline were observed in 450 µg ∙ g-1 Cd treated plants, Cd did not lead to a significant change in AsA, DHA and total AsA contents. According to the results of the research, E. sativa may be a Cd hyperaccumulator plant and we suggest that the plant may be a candidate plant for remediation of Cd-contaminated soil.

Early habituation of maize (Zea mays) suspension-cultured cells to 2,6-dichlorobenzonitrile is associated with the enhancement of antioxidant status.

The cellulose biosynthesis inhibitor 2,6-dichlorobenzonitrile (DCB) has been widely used to gain insights into cell wall composition and architecture. Studies of changes during early habituation to DCB can provide information on mechanisms that allow tolerance/habituation to DCB. In this context, maize-cultured cells with a reduced amount of cellulose (∼20%) were obtained by stepwise habituation to low DCB concentrations. The results reported here attempt to elucidate the putative role of an antioxidant strategy during incipient habituation. The short-term exposure to DCB of non-habituated maize-cultured cells induced a substantial increase in oxidative damage. Concomitantly, short-term treated cells presented an increase in class III peroxidase and glutathione S-transferase activities and total glutathione content. Maize cells habituated to 0.3-1 µM DCB (incipient habituation) were characterized by a reduction in the relative cell growth rate, an enhancement of ascorbate peroxidase and class III peroxidase activities, and a net increment in total glutathione content. Moreover, these cell lines showed increased levels of glutathione S-transferase activity. Changes in antioxidant/conjugation status enabled 0.3 and 0.5 µM DCB-habituated cells to control lipid peroxidation levels, but this was not the case of maize cells habituated to 1 µM DCB, which despite showing an increased antioxidant capacity were not capable of reducing the oxidative damage to control levels. The results reported here confirm that exposure and incipient habituation of maize cells to DCB are associated with an enhancement in antioxidant/conjugation activities which could play a role in incipient DCB habituation of maize-cultured cells.

Phosphorus improves arsenic phytoremediation by Anadenanthera peregrina by alleviating induced oxidative stress.

Due to similarities in their chemical behaviors, studies examining interactions between arsenic (As)--in special arsenate--and phosphorus (P) are important for better understanding arsenate uptake, toxicity, and accumulation in plants. We evaluated the effects of phosphate addition on plant biomass and on arsenate and phosphate uptake by Anadenanthera peregrina, an important Brazilian savanna legume. Plants were grown for 35 days in substrates that received combinations of 0, 10, 50, and 100 mg kg(-1) arsenate and 0, 200, and 400 mg kg(-1) phosphate. The addition of P increased the arsenic-phytoremediation capacity of A. peregrina by increasing As accumulation, while also alleviating As-induced oxidative stress. Arsenate phytotoxicity in A. peregrina is due to lipid peroxidation, but not hydrogen peroxide accumulation. Added P also increased the activity of important reactive oxygen species-scavenging enzymes (catalase and ascorbate peroxidase) that help prevent lipid peroxidation in leaves. Our findings suggest that applying P represents a feasible strategy for more efficient As phytoremediation using A. peregrina.

Effect of exogenous abscisic acid on the level of antioxidants in Atractylodes macrocephala Koidz under lead stress.

This study hypothesized that the positive or negative effects of exogenous abscisic acid (ABA) on oxidative stress caused by lead were dose dependent. The effects of different levels of ABA (2.5, 5, and 10 mg L(-1)) on lead toxicity in the leaves of Atractylodes macrocephala were studied by investigating plant growth, soluble sugars, proteins, lipid peroxidation, and antioxidative enzymes. Excess Pb inhibited root dry weight, root length, and the number of lateral roots, but increased shoot growth. In addition, lead stress significantly decreased the levels of chlorophyll pigments, protein, and activities of superoxide dismutase (SOD), catalase (CAT), ascorbate peroxidase (APX), and peroxidase (POD). Different levels of ABA significantly increased SOD, CAT, POD, and APX activities, but decreased the level of hydrogen peroxide and malondialdehyde in nonstressed plants. Exogenous application of 2.5 mg L(-1) ABA detoxified the stress-generated damages caused by Pb and also enhanced plant growth, soluble sugars, proteins, and all four antioxidant enzyme activities but reduced Pb uptake of lead-stressed plant compared to lead treatment alone. However, the toxic effects of Pb were further increased by the applications of 5 and 10 mg L(-1) ABA. The levels of antioxidants caused by a low concentration of exogenous ABA might be responsible for minimizing the Pb-induced toxicity in A. macrocephala.

Reactive oxygen species from chloroplasts contribute to 3-acetyl-5-isopropyltetramic acid-induced leaf necrosis of Arabidopsis thaliana.

3-Acetyl-5-isopropyltetramic acid (3-AIPTA), a derivate of tetramic acid, is responsible for brown leaf-spot disease in many plants and often kills seedlings of both mono- and dicotyledonous plants. To further elucidate the mode of action of 3-AIPTA, during 3-AIPTA-induced cell necrosis, a series of experiments were performed to assess the role of reactive oxygen species (ROS) in this process. When Arabidopsis thaliana leaves were incubated with 3-AIPTA, photosystem II (PSII) electron transport beyond Q(A) (the primary plastoquinone acceptor of PSII) and the reduction of the end acceptors at the PSI acceptor side were inhibited; this was followed by increase in charge recombination and electron leakage to O(2), resulting in chloroplast-derived oxidative burst. Furthermore, the main antioxidant enzymes such as superoxide dismutase (SOD), catalase (CAT) and ascorbate peroxidase (APX) lost their activity. Excess ROS molecules directly attacked a variety of cellular components and subsequently caused electrolyte leakage, lipid peroxidation and cell membrane disruption. Finally, this led to cell destruction and leaf tissue necrosis. Thus, 3-AIPTA-triggered leaf necrosis of Arabidopsis was found to be a result of direct oxidative injury from the chloroplast-originated ROS burst initiated by the inhibition of normal photosynthetic electron transport.

Effects of lead on the growth, lead accumulation and physiological responses of Pluchea sagittalis.

This work aimed to study the process of stress adaptation in root and leaves of different developmental stages (apex, middle and basal regions) of Pluchea sagittalis (Lam.) Cabrera plants grown under exposure to five Pb levels (0, 200, 400, 600 and 1000 µM) for 30 days. Pb concentration and content in roots, stems, and leaves of different developmental stages increased with external Pb level. Consumption of nutrient solution, transpiration ratio, leaf fresh weight, leaf area, and shoot length decreased upon addition of Pb treatments. However, dry weight of shoot parts and roots did not decrease upon addition of Pb treatments. Based on index of tolerance, the roots were much more tolerant to Pb than shoots. δ-aminolevulinic acid dehydratase activity was decreased by Pb treatments, whereas carotenoid and chlorophyll concentrations were not affected. Lipid peroxidation and hydrogen peroxide concentration both in roots and leaves increased with increasing Pb levels. Pb treatments increased ascorbate peroxidase activity in all plant parts, while superoxide dismutase activity increased in leaves and did not change in roots. Catalase activity in leaves from the apex shoot was not affected by Pb, but in other plant parts it was increased. Pb toxicity caused increase in non-protein thiol groups concentration in shoot parts, whereas no significant difference was observed in roots. Both root and shoot ascorbic acid concentration increased with increasing Pb level. Therefore, it seems that Pb stress triggered an efficient defense mechanism against oxidative stress in P. sagittalis but its magnitude was depending on the plant organ and of their physiological status. In addition, these results suggest that P. sagittalis is Pb-tolerant. In conclusion, P. sagittalis is able to accumulate on average 6730 and 550 µg Pb g(-1) dry weight, respectively, in the roots and shoot, a physiological trait which may be exploited for the phytoremediation of contaminated soils and waters.

Aluminum-induced oxidative stress and changes in antioxidant defenses in the roots of rice varieties differing in Al tolerance.

The effects of aluminum (Al) on root elongation, lipid peroxidation, hydrogen peroxide (H(2)O(2)) accumulation, antioxidant levels, antioxidant enzymatic activity, and lignin content in the roots of the Al-tolerant rice variety azucena and the Al-sensitive variety IR64 were investigated. Treatment with Al induced a greater decrease in root elongation and a greater increase in H(2)O(2) and lipid peroxidation as determined by the total thiobarbituric acid-reactive substance (TBARS) level in IR64 than in azucena. Azucena had significantly higher levels of superoxide dismutase, ascorbate peroxidase, glutathione reductase, and glutathione peroxidase GSH POD activity compared with IR64. The concentrations of reduced glutathione (GSH) and ascorbic acid, and the GSH/GSSG ratio (reduced vs. oxidized glutathione) were also higher in azucena than in IR64 in the presence of Al. The addition of 1 mg/L GSH improved root elongation in both varieties and decreased H(2)O(2) production under Al stress. By contrast, treatment with buthionine sulfoximine, a specific inhibitor of GSH synthesis, decreased root elongation in azucena and stimulated H(2)O(2) production in both varieties. Moreover, Al treatment significantly increased the cytoplasmic activity of peroxidase (POD) as well as the levels of POD bound ionically and covalently to cell walls in the Al-sensitive variety. The lignin content was also increased. Treatment with exogenous H(2)O(2) also increased the lignin content and decreased root elongation in IR64. These results suggest that Al induces lignification in the roots of Al-sensitive rice varieties, probably through an increase in H(2)O(2) accumulation.

Abscisic acid-regulated responses of aba2-1 under osmotic stress: the abscisic acid-inducible antioxidant defence system and reactive oxygen species production.

We investigated the interaction among abscisic acid (ABA), reactive oxygen species (ROS) and antioxidant defence system in the transduction of osmotic stress signalling using Arabidopsis thaliana WT (Columbia ecotype, WT) and an ABA-deficient mutant (aba2-1). For this, 50 µm ABA and osmotic stress, induced with 40% (w/v) polyethylene glycol (PEG8000; -0.7 MPa), were applied to WT and aba2-1 for 6, 12 or 24 h. Time course analysis was undertaken for determination of total/isoenzyme activity of the antioxidant enzymes, superoxide dismutase (SOD; EC 1.15.1.1), catalase (CAT; EC 1.11.1.6), ascorbate peroxidase (APX; EC 1.11.1.11), NADPH oxidase (NOX; EC 1.6.3.1) activity; scavenging activity of the hydroxyl radical (OH˙), hydrogen peroxide (H(2) O(2) ); endogenous ABA and malondialdehyde (MDA). The highest H(2) O(2) and MDA content was found in PEG-treated groups of both genotypes, but with more in aba2-1. ABA treatment under stress reduced the accumulation of H(2) O(2) and MDA, while it promoted activity of SOD, CAT and APX. APX activity was higher than CAT activity in ABA-treated WT and aba2-1, indicating a protective role of APX rather than CAT during osmotic stress-induced oxidative damage. Treatment with ABA also significantly induced increased NOX activity. Oxidative damage was lower in ABA-treated seedlings of both genotypes, which was associated with greater activity of SOD (Mn-SOD1 and 2 and Fe-SOD isoenzymes), CAT and APX in these seedlings after 24 h of stress. These results suggest that osmotic stress effects were overcome by ABA treatment because of increased SOD, CAT, APX and NOX.

Hexaconazole induces antioxidant protection and apigenin-7-glucoside accumulation in Matricaria chamomilla plants subjected to drought stress.

In this experiment, the possibility of enhancing the water deficit stress tolerance of chamomile (Matricaria chamomilla L.) during two growth stages by the exogenous application of hexaconazole (HEX) was investigated. To improve water deficit tolerance, HEX was applied in three concentrations during two different stages (50 and 80 days after sowing). After HEX applications, the plants were subjected to water deficit stress. Although all HEX concentrations improved the water deficit stress tolerance in chamomile plants, the application of 15 mg L(-1) provided better protection when compared to the other concentration. The exogenous application of HEX provided significant protection against water deficit stress compared to non-HEX-treated plants, significantly affecting the morphological characteristics and aspects of productivity, the relative water, protein and proline contents; non-enzymatic and enzymatic antioxidants; and the flower's apigenin-7-glucoside content. These results suggest that the HEX-induced tolerance to water deficit stress in chamomile was related to the changes in growth variables, antioxidants and the apigenin-7-glucoside content.

Salicylic acid alleviates decreases in photosynthesis under salt stress by enhancing nitrogen and sulfur assimilation and antioxidant metabolism differentially in two mungbean cultivars.

Salicylic acid (SA) is known to affect photosynthesis under normal conditions and induces tolerance in plants to biotic and abiotic stresses through influencing physiological processes. In this study, physiological processes were compared in salt-tolerant (Pusa Vishal) and salt-sensitive (T44) cultivars of mungbean and examined how much these processes were induced by SA treatment to alleviate decrease in photosynthesis under salt stress. Cultivar T44 accumulated higher leaf Na(+) and Cl(-) content and exhibited greater oxidative stress than Pusa Vishal. Activity of antioxidant enzymes, ascorbate peroxidase (APX) and glutathione reductase (GR) was greater in Pusa Vishal than T44. Contrarily, activity of superoxide dismutase (SOD) was greater in T44. The greater accumulation of leaf nitrogen and sulfur through higher activity of their assimilating enzymes, nitrate reductase (NR) and ATP-sulfurylase (ATPS) increased reduced glutathione (GSH) content more conspicuously in Pusa Vishal than T44. Application of 0.5 mM SA increased nitrogen and sulfur assimilation, GSH content and activity of APX and GR. This resulted in the increase in photosynthesis under non-saline condition and alleviated the decrease in photosynthesis under salt stress. It also helped in restricting Na(+) and Cl(-) content in leaf, and maintaining higher efficiency of PSII, photosynthetic N-use efficiency (NUE) and water relations in Pusa Vishal. However, application of 1.0 mM SA resulted in inhibitory effects. The effect of SA was more pronounced in Pusa Vishal than T44. These results indicate that SA application alleviates the salt-induced decrease in photosynthesis mainly through inducing the activity of NR and ATPS, and increasing antioxidant metabolism to a greater extent in Pusa Vishal than T44.

Heme oxygenase-1 is involved in the cytokinin-induced alleviation of senescence in detached wheat leaves during dark incubation.

This study tested whether an inducible isoform of heme oxygenase (HO, EC 1.14.99.3), HO-1, is involved in the cytokinin (CTK)-induced alleviation of senescence in detached wheat (Triticum aestivum L.) leaves during dark incubation. We discovered that exogenous supplement of 6-benzylaminopurine (6-BA) at 10 µM for 48 h not only delayed the dark-induced loss of chlorophyll and protein contents in detached wheat leaves, but also significantly increased HO activity in a time-dependent manner. This induction reached a maximum within 3h of 6-BA supply, which was further confirmed by using semi-quantitative RT-PCR and protein gel blot analysis. Furthermore, the decreases in intracellular thiobarbituric acid reactive substances (TBARS) content, and the increases in the transcript level, total and isozymatic activities of some important antioxidant enzymes, such as catalase (CAT, EC 1.11.1.6), peroxidase (POD, EC 1.11.1.7), superoxide dismutase (SOD, EC 1.15.1.1), and ascorbate peroxidase (APX, EC 1.11.1.11), were observed. Reversed responses of chlorophyll, protein and TBARS contents, HO activity, and the expression of above antioxidant enzymes were observed when zinc protoporphyrin-IX (ZnPPIX), a potent HO-1 inhibitor, was added together with 6-BA. In contrast, HO-1 inducer hemin could partially mimic the effects of 6-BA. Together, the results suggest that HO-1 might be involved in the CTK-induced alleviation of senescence and lipid peroxidation in detached wheat leaves.