High plasma thiocyanate levels are associated with enhanced myeloperoxidase-induced thiol oxidation and long-term survival in subjects following a first myocardial infarction

Abstract

Elevated levels of myeloperoxidase (MPO) are associated with poor cardiovascular outcomes. MPO uses H₂O₂ to generate oxidants including HOCl and HOSCN, from chloride and thiocyanate (SCN^?) ions, respectively. SCN^? is the preferred substrate. Elevation of this anion decreases HOCl generation and increases HOSCN formation, a thiol-specific oxidant. Such changes are of potential relevance to people with elevated SCN^? levels, such as smokers. In this retrospective study, we examined whether elevated plasma MPO and SCN^? levels increased thiol oxidation as a result of increased HOSCN formation, and impacted on long-term survival in 176 subjects (74 non-smokers, 46 smokers, and 56 previous smokers) hospitalized after a first myocardial infarction. Plasma thiols were not significantly altered in smokers compared to non-smokers or past smokers. However, significant positive correlations were detected between SCN^? levels and MPO-induced thiol loss in the total population (r = 0.19, P = 0.020) and smokers alone (r = 0.58, P < 0.0001). Twelve-year all-cause mortality data indicate that above median MPO is significantly associated with higher mortality, but below-median MPO and above-median SCN^? results in increased survival, compared to below-median SCN^?. Cox proportional hazard analysis showed a significant decrease in mortality for each 1 μM increase in SCN^? (0.991; P = 0.040). Subject age was, as expected, a strong predictor of subject survival. Overall these data suggest that subjects with below-median MPO and above-median SCN^? have better long-term survival, and that elevated plasma levels of SCN^? might be protective in at least some populations.     

黄芩黄酮对硒性白内障晶状体抗氧化酶表达的影响

为探讨黄芩黄酮防治白内障的作用机理 ,采用半定量RT PCR方法比较正常组、白内障组和中药防治组大鼠晶状体中GSH Px、GR和Cu ZnSOD的mRNA水平 .白内障组GSH Px、GR和Cu ZnSOD的mRNA水平在 15d龄时显著高于正常 ,然后下降 ;在 2 7d和 31d龄 ,GR和Cu ZnSOD的mRNA水平下降至与正常无显著差异 ,GSH PxmRNA水平仍略高于正常 .中药防治组晶状体中 ,3种抗氧化酶的mRNA水平在各实验取样点无明显变化 ;其中 ,GR和Cu ZnSOD的mRNA水平一直与正常无显著差异 ,GSH PxmRNA水平略高于正常 .黄芩黄酮可能通过有效清除亚硒酸钠间接产生的活性氧来防止白内障的发生 ,并使亚硒酸钠对晶状体抗氧化酶表达的影响得以消除     

Erdosteine modulates radiocontrast‐induced hepatotoxicity in rat

It has been suggested that reactive oxygen species (ROS) plays an important role in radio contrast media (RCM)‐induced ischemia reperfusion tissue injury although antioxidants may have protective effects on the injury. We investigated the effects of erdosteine as an antioxidant agent on RCM‐induced liver toxicity in rats by evaluation of lipid peroxidation (as TBARS), catalase (CAT), superoxide dismutase (SOD), reduced glutathione (GSH) and glutathione peroxidase (GSH‐Px) values and histological evaluation. Twenty‐one rats were equally divided into three groups as follows: control, RCM, and RCM plus erdosteine. RCM was intraperitoneally administered for 1 day. Erdosteine was administered orally for 2 days after RCM administration. Liver samples were taken from the rats and they homogenized in a motor‐driven tissue homogenizer. TBARS levels were significantly (p < 0.005) higher in RCM group than in control although SOD activities significantly (p < 0.05) decreased in RCM group. TBARS levels were lower in RCM plus erdosteine group than in control although SOD activity and GSH level increased (p < 0.05) in liver as compared to RCM alone. Erdosteine showed also histopathological protection (p < 0.0001) against RCM induced hepatotoxicity. GSH‐Px and CAT activities were not statistically changed by the erdosteine. According to our results, it can be concluded that radiocontrast media can induce oxidative stress in liver as suggested by previous studies. Erdosteine seems to be protective agent on the radiocontrast media‐induced liver toxicity by inhibiting the production of ROS via the enzymatic antioxidant system. Copyright © 2009 John Wiley & Sons, Ltd.     

Zinc nutrition increases the antioxidant defenses of honey bees

Is the typical zinc (Zn) content of honey and pollen sufficient to meet the nutritional requirements of honey bees? To answer this question, and find the optimal dietary Zn levels for honey bees, we investigated the effects of varying dietary Zn levels on both captive worker bees and free‐flying honey bees, Apis mellifera ligustica Spinola (Hymenoptera: Apidae). We fed captive workers and free‐flying honey bees with 50% (wt/wt) sucrose solutions with Zn levels of either 0, 15, 30, 45, 60, or 75 mg kg^?1 diet and measured their Cu/Zn‐SOD activity, the mean survival time of captive bees, the Cu/Zn‐SOD activity of larvae, and the Zn concentration of royal jelly. Captive workers provided with 30 mg kg^?1 dietary Zn had higher Cu/Zn‐SOD activity and mean survival time than the control. Dietary Zn levels from 60 to 75 mg kg^?1 significantly increased the Zn content of royal jelly provided by colonies and the Cu/Zn‐SOD activity of larvae. Honey or pollen with a Zn content of <30 mg kg^?1 was insufficient to satisfy the maintenance nutritional requirements of bees that were not raising larvae. It therefore seems advisable to supply supplementary Zn to non‐brooding colonies when the Zn content of honey or pollen is <30 mg kg^?1. Honey or pollen with a Zn content of 60 mg kg^?1 was sufficient to satisfy the nutritional requirements for royal jelly production and to improve the health of larvae. It may therefore also be advisable to provide supplementary Zn to colonies with larvae when the Zn content of honey or pollen is <60 mg kg^?1.     

Brassica rapa Polysaccharides Ameliorate CCl4‐Induced Acute Liver Injury in Mice through Inhibiting Inflammatory Apoptotic Response and Oxidative Stress

Brassica rapa L., also called NIUMA, is used empirically in Tibetan medicine for its antioxidant, anti‐inflammatory and antiradiation activities. This study explored the hepatoprotective effects of B. rapa polysaccharides (BRPs) on acute liver injury induced by carbon tetrachloride (CCl₄) in mice and the underlying mechanisms. Mice were treated with CCl₄ after the oral administration of BRPs (55, 110 and 220 mg/kg) or bifendate (100 mg/kg) for 7 days. Blood and liver samples of mice were collected for analysis after 24 h. The ALP, ALT and AST levels and the biological activities of SOD, MDA and GSH?Px were measured. Histopathological changes in the liver were determined through hematoxylin and eosin staining. Moreover, TNF‐α, IL‐1β and IL‐6 expression levels were detected by commercial reagent kits. Finally, Western blot analysis was used to check the relative expression levels of caspase‐3, p‐JAK2 and p‐STAT3. The BRP pre‐treatment significantly decreased the enzymatic activities of ALT, ALP and AST in the serum, markedly increased the activities of SOD and GSH?Px in the liver and reduced the MDA concentration in the liver. BRPs alleviated hepatocyte injury and markedly inhibited the expression of TNF‐α, IL‐1β and IL‐6, also downregulating the CCl₄‐induced hepatic tissue expression of caspase‐3. Furthermore, BRPs inhibited the JAK2/STAT3 signaling pathway in a dose‐dependent manner in the liver. This study demonstrated that BRPs exert hepatoprotective effect against the CCl₄‐induced liver injury via modulating the apoptotic and inflammatory responses and downregulating the JAK2/STAT3 signaling pathway. Therefore, B. rapa could be considered a hepatoprotective medicine.     

Complexation of a Ligand with a Surfactant Micelle for Soil Heavy Metal Desorption

The ligands iodide (I^?) and thiocyanate (SCN^?), alone or in admixture, in combination with a non-ionic surfactant, Triton X-100, were evaluated as washing agents for heavy metal desorption from a contaminated soil. After seven successive washings, selective sequential extraction (SSE) was performed to determine the heavy metal content that remained associated with each geo-chemical fraction of the soil. The surfactant with 0.336 mol L^?1 of ligand I^? removed 75% Cd and 23% Cu, whereas the mobilization of Zn and Pb were not significant after 7 washings. At a concentration of 0.286 mol L^?1, the ligand SCN^? in the presence of surfactant removed 36% Cd, 44% Cu and 77% Zn. Among the washing agents, the combination of I^? and SCN^? produced the highest desorption efficiencies 95% Cd, 48% Cu, and 3.1% Pb, but not for Zn. The SCN^? ligand extracted the most Zn (77%). The SSE procedure indicated that the I^? removed metals from the exchangeable, carbonate and oxide fractions whereas SCN^? removed metals only from the exchangeable fraction. Both ligands, in the presence of surfactant, removed Cu from all fractions except the exchangeable sites, whereas only SCN^? plus surfactant removed Zn from all fractions. The ligand mixture plus surfactant mobilized only limited quantities of Pb from the oxide and residual fractions.     

Neuroprotective Effects of Butanol Fraction of Cordyceps cicadae on Glutamate‐Induced Damage in PC12 Cells Involving Oxidative Toxicity

The current study was aimed at investigating the neuroprotective effects of the butanol fraction from Cordyceps cicadae (C_BU), which was responsible for the anti‐aging effect of this medicine. Glutamate‐induced PC12 cells were used as a model to determine the neuroprotective effect against oxidative cell death. Cell viability, cytotoxicity, flow cytometry, mitochondrial transmembrane potential (MMP), reactive oxygen species (ROS), glutathione peroxidase (GSH‐Px), and superoxide dismutase (SOD) levels were analyzed to assess neuronal cell survival or death. The results obtained from the above evaluations showed that C_BU was the most effective fraction and even better than pure compounds present in C. cicadae in terms of suppressing glutamate‐induced damage in PC12 cells, increasing cell viability, decreasing lactase dehydrogenase (LDH) release, and reduction of apoptosis induced by exposure to glutamate. Furthermore, C_BU protected cells against mitochondrial dysfunction and oxidative stress as indicated by the suppression of ROS accumulation and up regulation of the levels of GSH‐Px and SOD. In summary, the above results showed that C_BU exerted neuroprotective effect against oxidative damage, and this activity could be partly due to the action of nucleosides present in the C_BU.     

Influence of smoking on serum and milk malondialdehyde, superoxide dismutase, glutathione peroxidase, and antioxidant potential levels in mothers at the postpartum seventh day

The aim of the study was to investigate simultaneously serum and milk malondialdehyde (MDA) levels, superoxide dismutase (SOD), glutathione peroxidase (GPx) activities, and antioxidant potential (AOP) in active-smoking, passive-smoking, and nonsmoking mothers and to search if there is any difference between serum and milk oxidant/ antioxidant status caused by smoking. According to their smoking status, 60 mothers (age range: 20–35 yr) were classified into one of three groups: the active-smoking mothers (n=15), the passive-smoking mothers (n=22), and the nonsmoking mothers (n=23). Serum and milk MDA, SOD, GPx, and AOP values were determined in mothers on the postpartum seventh day by the spectrophotometric method. Serum Zn and Cu concentrations were determined by atomic absorption spectrophotometry (AAS). There was no significant difference in serum samples with respect to MDA (p=0.17), SOD (p=0.51) and AOP (p=0.36) levels, but there was a significant difference in serum GPx (p=0.002) levels among the study groups. The significant differences were also found in milk samples in terms of MDA (p=0.002) and SOD (p=0.011), but not in GPx (p=0.11) and AOP (p=0.29) levels among the study groups. No significant difference was seen in serum zinc concentration (p=0.49), but copper concentration differed significantly among the groups (p=0.005). These observations suggest that human milk is more vulnerable to oxidative stress and lipid peroxidation than serum samples in smoking mothers, even if they are passive smokers.     

New volatile anesthetic,desflurane, reduces vitamin E level in blood of operative patients via oxidative stress

It has been well known that some volatile anesthetic agents produce oxidative stress. Desflurane as a new volatile agent might have limited oxidative toxic effect because it is relatively a new short‐acting anesthetic characterized by a short duration of action and a quick postanesthetic recovery. We investigated effect of desflurane on serum glutathione peroxidase (GSH‐Px), lipid peroxidation (LP), vitamin E, and erythrocyte superoxide dismutase (SOD) values in patients. Fifteen adult patients are scheduled for elective surgery, ASA I or II physical status. Tidal volume and ventilation frequency were kept unchanged during the operation. Baseline values in venous blood samples were preoperatively taken and blood was also taken postoperatively at the 1st and the 12th hours of desflurane exposure. LP levels were significantly (p < 0.05) higher postoperatively at 1st hour than in preoperative values while α‐tocopherol concentration was significantly (p < 0.001) lower in postoperative period at 1st hour than in preoperative period. Erythrocyte SOD and serum GSH‐Px activities did not differ between pre‐ and postoperative periods. In conclusion, we observed that desflurane produced oxidative stress by decreasing α‐tocopherol levels. Use of vitamin E may be possible to reduce the oxidative effect of desflurane. Copyright © 2010 John Wiley & Sons, Ltd.     

Protective effects of ascorbic acid on hepatotoxicity and oxidative stress caused by carbon tetrachloride in the liver of Wistar rats

This study was planned to investigate the protective effect of l (+)‐ascorbic acid (Vit C) on CCl₄‐induced hepatotoxicity and oxidative stress in the liver of Wistar rats (Rattus Norvegicus, strain Wistar). Twenty‐four adult male Wistar rats were fed with standard rat chow diet for 10 days and randomly were divided into four groups of six each as follows: (1) control, (2) CCl₄, (3) “CCl₄ + Vit C”, (4) Vit C groups. CCl₄ was applied to rats belonging to CCl₄ and “CCl₄ + Vit C” groups subcutaneously at 1 mg kg^?1 dose CCl₄ for 3 days. Vit C applied to “CCl₄ + Vit C” and “Vit C” group rats intraperitoneally at 300 mg kg^?1 dose for 3 days. All rats were sacrificed and livers were quickly removed on the fourth day of the experiment. MDA, total glutathione (T.GSH) levels and superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GSH‐PX) activities were measured in the liver of all groups of rats and also serum alanine amino transferase (ALT) and aspartate amino transferase (AST) activities were detected to determine liver functions in all groups of rats. Histopathological changes were evaluated by light and transmission electron microscopes. In “CCl₄ + Vit C” group, MDA level was significantly decreased (p < 0.05) and SOD, CAT, GSH‐PX activities were significantly increased (p < 0.005, 0.01, 0.05) respectively, T.GSH level was significantly increased (p < 0.005) and serum ALT and AST activities were significantly decreased (p < 0.01, 0.05), respectively, when compared with CCl₄ group. These results show that Vit C has a highly protective effect on hepatotoxicity and oxidative stress caused by CCl₄. Copyright © 2009 John Wiley & Sons, Ltd.     

Increased nitration and diminished activity of copper/zinc superoxide dismutase in placentas from diabetic rats

Abstract

Nitration-induced protein damage in the placenta leads to impaired blood flow and deficient feto–placental exchange in diabetic pregnancies. This work studied the effect of nitric oxide and peroxynitrite on Cu/Zn SOD activity in rat placentas and evaluated whether Cu/Zn SOD is nitrated in the placenta from diabetic rats at mid-gestation. Protein nitration was evaluated by EIA, Cu/Zn SOD activity by inhibition of the epinephrine auto-oxidation, Cu/Zn SOD expression by western blot and specific nitration by immunoprecipitation. This study found higher levels of protein nitration (p < 0.001), diminished Cu/Zn SOD activity and enhanced protein expression (p < 0.01) in placentas from diabetic rats. Placental Cu/Zn SOD activity was inhibited by peroxynitrite (p < 0.01). Besides, nitration of Cu/Zn SOD was elevated in placentas from diabetic rats (p < 0.01). These results show that rat Cu/Zn SOD can be nitrated, a modification that could lead to the depressed activity of this enzyme found in placentas from diabetic rats.     

紫外线对大鼠晶状体抗氧化酶mRNA表达水平的影响

为探讨紫外线对晶状体的损伤机制,用ＲＴ－ＰＣＲ方法（ｒｅｖｅｒｓｅｔｒａｎｓｃｒｉｐｔｉｏｎ－ｐｏｌｙｍｅｒａｓｅｃｈａｉｎｒｅａｃｔｉｏｎ,反转录聚合酶链反应）,研究经紫外线照射后大鼠晶状体抗氧化相关酶,包括铜锌－超氧化物歧化酶（ｃｏｐｐｅｒ－ｚｉｎｃ－ｓｕｐｅｒｏｘｉｄｅｄｉｓｍｕｔａｓｅ,Ｃｕ－Ｚｎ－ＳＯＤ）,谷胱甘肽过氧化物酶（ｇｌｕ－ｔａｔｈｉｏｎｅｐｅｒｏｘｉｄａｓｅ,ＧＳＨ－Ｐｘ）和过氧化氢酶（ｃａｔａｌａｓｅ,ＣＡＴ）等ｍＲＮＡ的表达．结果显示,短时间的照射（２～５ｍｉｎ）,抗氧化相关酶的ｍＲＮＡ表达水平有增高表现,随后其ｍＲＮＡ表达水平开始下降,１５ｍｉｎ时抗氧化相关酶ｍＲＮＡ的表达下降更为明显,与对照组相比有非常显著性差异（Ｐ＜０．００１）．照射后２４ｈ,抗氧化相关酶的ｍＲＮＡ表达有不同程度的恢复;照射后４８ｈ,其ｍＲＮＡ表达水平基本恢复,与对照组相比没有显著性差异．从而从基因水平上初步探讨了紫外线的氧化损伤机制     

Protective mechanisms of Pycnogenol® in ethanol‐insulted cerebellar granule cells

Pycnogenol® (PYC), a patented combination of bioflavonoids extracted from the bark of French maritime pine (Pinus maritima), inhibits apoptosis and necrosis of developing neurons exposed acutely to ethanol (EtOH). The present study shows that the protective mechanisms of PYC in EtOH‐exposed postnatal day 9 cerebellar granule cells (P9 CGCs) include (1) reduction of reactive oxygen species (ROS) production; (2) counteraction of suppressed copper/zinc superoxide dismutase (Cu/Zn SOD) and glutathione peroxidase/reductase (GSH‐Px/GSSG‐R) system activities; (3) upregulation of Cu/Zn SOD protein expression; (4) mitigation of the EtOH‐mediated exacerbation of catalase (CAT) activity; and, (5) specific binding and inhibition of active caspase‐3. These results indicate that the mechanisms by which PYC antagonizes EtOH‐induced oxidative stress include oxidant scavenging and modulation of endogenous, cellular proteins. Using findings from the present and previous studies, a model delineating the mechanisms of EtOH effects on the system of antioxidant enzymes in developing CGCs is presented. © 2004 Wiley Periodicals, Inc. J Neurobiol, 2004     

Effects of ammonia on growth,digestion and antioxidant capacity in juvenile yellow catfish Pelteobagrus fulvidraco (Richardson, 1846)

Effects of ammonia stress on food ingestion, growth, digestion and antioxidant capacity were investigated in juvenile yellow catfish Pelteobagrus fulvidraco (Richardson) with initial body weights of 20.24 ± 0.18 g. The fish were reared in triplicate in 15 experimental tanks at a rate of 30 fish per tank for 56 days. Water was maintained at a dissolved oxygen (DO) level of over 6.2 mg L^?1, pH 7.2–7.6, and temperature of 29.0 ± 1.5°C under a natural 12L: 12D photoperiod. Survival, food ingestion (FI), specific growth rate (SGR), food conversion efficiency (FCE), apparent digestibility coefficient (ADC), total antioxidant capacity (T‐AOC), levels of glutathione (GSH) and malonaldehyde (MDA), and activities of superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GSH‐PX) of the juveniles were determined in total ammonia nitrogen (NH₃‐N+NH₄‐N) levels of 0 (control group), 3.36, 6.72, 13.44 and 26.88 mg L^?1. The results show that the survival, FI, SGR, FCE, and ADC decreased significantly with an increase in total ammonia nitrogen (p < .05), and a significantly negative correlation between SGR and total ammonia nitrogen levels (p < .05). T‐AOC, SOD, CAT, GSH‐PX and GSH in the blood, liver and gills were found to decline significantly with an increase in the total ammonia nitrogen level (p < .05), while the MDA in the blood, liver and gills was elevated significantly with the increase in total ammonia nitrogen (p < .05). The results indicate a threshold in the induction of the T‐AOC and activities of antioxidant enzymes in yellow catfish tissues with a total ammonia nitrogen increase. In the present study the total ammonia nitrogen threshold thus changed from 6.72 mg L^?1 in the juvenile yellow catfish.     

Doxorubicin Resistance Conferred by Selective Enhancement of Intracellular Glutathione Peroxidase or Superoxide Dismutase Content in Human MCF-7 Breast Cancer Cells

To examine the role of doxorubicin-stimulated oxyradical formation in tumor cell killing, we introduced glutathione peroxidase (GSH Px) or superoxide dismutase (SOD) into MCF-7 cells by “scrape loading.” Control cytoplasmic GSH Px and SOD levels increased from (mean ± S.E.) 0.37nmol/min/mg and 0.58 μg SOD/mg, respectively, to 3.99 or 7.63 nmol/min/mg and 1.40 or 1.83 μg SOD/mg after treatment with either 150 or 300 units/ml of GSH Px or 20 or 40mg/ml SOD. Resistance to doxorubicin was cbrrelated with the level of GSH Px introduced into the MCF-7 cells: a one-hour exposure to 1.75 μM doxorubicin decreased the cloning efficiency of control cells loaded with medium alone to 34%, whereas doxorubicin-treated cells augmented with 150 or 300 units/ml of GSH Px had plating efficiencies of 56 or 86%, P < 0.05. Introduction of SOD increased MCF-7 resistance to doxorubicin similarly. The heat-inactivated enzymes were not protective. Cells loaded with GSH Px were also resistant to the redox cycling anticancer quinone mitomycin C but not to the redox inactive analogs 5-iminodaunorubicin and mitoxan-trone. suggesting that amplification of GSH Px or SOD levels can produce doxorubicin resistance in MCF-7 cells.     

The effect of elevated hydrostatic pressure upon selected biomarkers in juvenile blackspot seabream Pagellus bogaraveo in a 14 day‐long experiment

Hydrostatic pressure elevated to 500 kPa for 14 days was found to affect hepatic 7‐ethoxyresorufin‐O‐deethylase (EROD), oxidized protein (POx), protein yield and branchial Na⁺–K⁺‐ATPase. No effect on glutathione‐S‐transferase (GST), superoxidase dismutase (SOD), catalase (CAT), lipid peroxidation (LP), acetylcholinesterase (AChE), butyrylcholinesterase (BChE), condition factor (K) and hepato‐somatic index (I_H) was encountered.     

Copper Intoxication; Antioxidant Defenses and Oxidative Damage in Rat Brain

Copper (Cu) is an integral part of many important enzymes involved in a number of vital biological processes. Even though Cu is essential to life, it can become toxic to cells, at elevated tissue concentrations. Oxidative damage due to Cu has been reported in recent studies in various tissues. In this study, we aimed to determine the effect of excess Cu on oxidative and anti-oxidative substances in brain tissue in a rat model. Sixteen male Wistar albino rats were divided into two groups: the control group, which was given normal tap water, and the experimental group, which received water containing Cu in a dose of 1 g/l. All rats were sacrificed at the end of 4 wk, under ether anesthesia. Cu concentration in the liver and in plasma alanine aminotransferase (ALT) and aspartate transaminase (AST) activities were determined. There were multiparameter changes with significant ALT and AST activity elevation and increased liver Cu concentration. In brain tissue, Cu concentration, superoxide dismutase (SOD) activities, malondialdehyde (MDA) levels and glutathione (GSH) concentrations were determined. Brain Cu concentration was significantly higher in rats receiving excess Cu, compared with control rats (p < 0.05). Our results showed that SOD activities and GSH levels in brain tissue of the Cu-intoxicated animals were significantly lower than in the control group (p < 0.01 and p < 0,001, respectively). The brain MDA levels were found to be significantly higher in the experimental group than in the control group (p < 0.001). The present results indicate that excessive Cu accumulation in the brain depressed SOD activities and GSH levels and resulted in high MDA levels in brain homogenate due to the lipid peroxidation induced by the Cu overload.     

Effect of N‐acetylarginine,a metabolite accumulated in hyperargininemia,on parameters of oxidative stress in rats: protective role of vitamins and L‐NAME

In the present investigation, we initially evaluated the in vitro effect of N‐acetylarginine on thiobarbituric acid‐reactive substances (TBA‐RS), total sulfhydryl content and on the activities of antioxidant enzymes such as catalase (CAT), superoxide dismutase (SOD) and glutathione peroxidase (GSH‐Px) in the blood, kidney and liver of rats. Results showed that N‐acetylarginine, at a concentration of 5.0 μM, decreased the activity of CAT in erythrocytes, enhanced TBA‐RS in the renal cortex, decreased CAT and SOD activities in the renal medulla and decreased CAT and increased SOD and GSH‐Px activities in the liver of 60‐day‐old rats. Furthermore, we tested the influence of the antioxidants, trolox and ascorbic acid, as well as of the N^ω‐nitro‐L ‐arginine methyl ester (L‐NAME) on the effects elicited by N‐acetylarginine on the parameters tested. Antioxidants and L‐NAME prevented most of the alterations caused by N‐acetylarginine on the oxidative stress parameters evaluated. Data indicate that oxidative stress induction is probably mediated by the generation of NO and/or ONOO^? and other free radicals because L‐NAME and antioxidants prevented the effects caused by N‐acetylarginine in the blood, renal tissues and liver of rats. Our findings lend support to a potential therapeutic strategy for this condition, which may include the use of appropriate antioxidants for ameliorating the damage caused by N‐acetylarginine. Copyright © 2014 John Wiley & Sons, Ltd.     

Towards soil management with Zn and Mn: estimates of fertilisation efficacy of Citrus trees

Nutrient management recommendations for fruit crops lack the understanding of the efficiency of soil fertilisation with manganese (Mn) and zinc (Zn), which could substitute, in part, the traditional foliar applications. Fruit yield of trees in response to Zn and Mn supply via soil may be limited either by sorption reactions with soil colloids or low solubility of fertilisers. We investigated the effects of fertiliser sources and rates of Mn and Zn applied to soils with different sorption capacities on nutrient uptake, biochemical responses and biomass of Citrus. Two experiments were carried out with 2‐year‐old sweet orange trees that received applications of Mn or Zn. The first experiment evaluated the application of Mn fertilisers (MnCO₃ and MnSO₄) at three levels of the nutrient (0, 0.7 and 3.5 g plant^?1 of Mn) in two types of soil (18.1% and 64.4% of clay, referred to as sandy loam and clay soils, respectively). The second experiment, likewise, evaluated Zn fertilisers (ZnO and ZnSO₄) and nutrient levels (0, 1.0 and 5.0 g plant^?1 of Zn). Application of Mn and Zn increased nutrient availability in the soils as well as leaf nutrient concentrations in the trees. The lowest rates, 0.7 g plant^?1 of Mn and 1.0 g plant^?1 of Zn, both as sulphate, were sufficient to supply these micronutrients to sufficient levels in leaves, flowers and fruits. Metal toxicity to plants occurred with higher doses of both nutrients and to a large extent in the sandy soil. In this case, protein bands lower than 25 kDa were observed as well a decrease on leaf chlorophyll content. In the clay soil, despite increased micronutrient concentrations in the plant, responses were less pronounced because of higher adsorption of metals in the soil. Superoxide dismutase (SOD, EC 1.15.1.1) isoenzyme activity was determined by non‐denaturing polyacrylamide gel electrophoresis (PAGE). The Cu/Zn‐SOD isoenzymes increased with increased Zn rates, but in contrast, when Mn was applied at the highest rate, the activity of Cu/Zn‐SODs decreased. The SOD activity pattern observed indicated increased production of superoxide and consequently an oxidative stress condition at the highest rates of Zn and Mn applied. The results demonstrated that the soil application of Mn and Zn can supply nutrient demands of orange trees, however the low solubility of fertilisers and the high sorption capacity of soils limit fertilisation efficiency. On the contrary, application of sulphate source in sandy soils may cause excess uptake of Mn and Zn and oxidative stress, which impairs the photosynthetic apparatus and consequently tree growth.     

Involvement of glutathione metabolism in Eichhornia crassipes tolerance to arsenic

• Aquatic macrophytes are potentially useful for phytoremediation programmes in environments contaminated by arsenic (As). Biochemical and physiological modification analyses in different plant parts are important to understand As tolerance mechanisms.
• The objective was to evaluate glutathione metabolism in leaves and roots of Eichhornia crassipes (Mart.) Solms treated to As. Specimens of E. crassipes were cultured for 3 days in Clark's nutrient solution containing 7 μm As. The enzymes ATP sulphurylase (ATPS), glutathione reductase (GR), glutathione peroxidase (GSH‐Px), glutathione sulphotransferase (GST) and γ‐glutamylcysteine synthetase (γ‐ECS) activity, glutathione content, total protein and non‐protein thiols were evaluated.
• The ATPS activity increased in roots. GR activity in leaves and GSH‐Px in roots were lower. GST activity was higher in roots and lower in leaves, and γ‐ECS activity was higher in leaves. Glutathione levels were lower, total thiol levels were higher and non‐protein levels did not change in E. crassipes leaves and roots. Exposure to As increased enzyme activity involved with sulphur metabolism, such as ATPS. Higher GR activity and lower GSH‐Px indicate increased glutathione conjugation to As due to increased GSH availability. The higher GST activity indicates its participation in As detoxification and accumulation through As GSH conjugation. Changes in glutathione and thiol levels suggest high phytochelatin synthesis.
• In conclusion, the increments in ATPS, GR, GST and γ‐ECS activity indicate that these enzymes are involved in GSH metabolism and are part of the E. crassipes As detoxification mechanism.