Luciane Belmonte Pereira

Cadmium and mineral nutrient accumulation in potato plantlets grown under cadmium stress in two different experimental culture conditions

In order to evaluate the effect of cadmium (Cd(2+)) toxicity on mineral nutrient accumulation in potato (Solanum tuberosum L.), two cultivars named Asterix and Macaca were cultivated both in vitro and in hydroponic experiments under increasing levels of Cd(2+) (0, 100, 200, 300, 400 and 500 microM in vitro and 0, 50, 100, 150 and 200 microM in hydroponic culture). At 22 and 7 days of exposure to Cd(2+), for the in vitro and hydroponic experiment, respectively, the plantlets were separated into roots and shoot, which were analyzed for biomass as well as Cd(2+), and macro (Ca(2+), K(+) and Mg(2+)) and micronutrient (Cu(2+), Fe(2+), Mn(2+) and Zn(2+)) contents. In the hydroponic experiment, there was no reduction in shoot and root dry weight for any Cd(2+) level, regardless of the potato cultivar. In contrast, in the in vitro experiment, there was an increase in biomass at low Cd(2+) levels, while higher Cd(2+) levels caused a decrease. In general, Cd(2+) decreased the macronutrient and micronutrient contents in the in vitro cultured plantlets in both roots and shoot of cultivars. In contrast, the macronutrient and micronutrient contents in the hydroponically grown plantlets were generally not affected by Cd(2+). Our data suggest that the influence of Cd(2+) on nutrient content in potato was related to the level of Cd(2+) in the substrate, potato cultivar, plant organ, essential element, growth medium and exposure time.

Cadmium toxicity causes oxidative stress and induces response of the antioxidant system in cucumber seedlings

In this study, the effects of cadmium (Cd) on lipid peroxidation, electrolyte leakage, protein oxidation, ascorbate peroxidase (APX; E.C. 1.11.1.11), catalase (CAT; E.C. 1.11.1.6) and superoxide dismutase (SOD; E.C. 1.15.1.1) activities, and ascorbic acid, non-protein thiol groups and total soluble protein contents in cucumber seedlings (Cucumis sativus L.) were investigated. Seedlings were grown in vitro in an agar-solidified substrate containing four Cd levels as CdCl2 (0, 100, 400, and 1000 µmol L-1) for 10 d. The lowest Cd level decreased the malondialdehyde concentration. Electrolyte leakage increased only at 1000 µmol Cd L-1, whereas protein oxidation and total soluble protein content were enhanced at 400 and 1000 µmol Cd L-1. Activity of APX was inhibited while the activities of CAT and SOD were increased at all Cd concentrations. Ascorbic acid was enhanced at 400 and 1000 µmol Cd L-1 whereas non-protein thiol groups were increased at all Cd supplies. The results evidence the importance of the enzymatic and non-enzymatic antioxidant system in response to cadmium toxicity in cucumber seedlings.

Oxidative stress is an early symptom triggered by aluminum in Al-sensitive potato plantlets.

The objective of this study was to evaluate whether the oxidative stress caused by aluminum (Al) toxicity is an early symptom that can trigger root growth inhibition in Macaca (Al-sensitive) and SMIC148-A (Al-tolerant) potato clones. Plantlets were grown in a nutrient solution (pH 4.00) with 0, 100 and 200mg Al L(-1). At 24, 72, 120 and 168h after Al addition, root length and biochemical parameters were determined. Regardless of exposure time, root length of the Macaca clone was significantly lower at 200mg Al L(-1). For the SMIC148-A clone, root length did not decrease with any Al treatments. Al supply caused lipid peroxidation only in Macaca, in both roots (at 24, 72, 120 and 168h) and shoot (at 120 and 168h). In roots of the Macaca, catalase (CAT) and ascorbate peroxidase (APX) activity decreased at 72 and 120h, and at 24, 72 and 120h, respectively. At 168h, both activities increased upon addition of Al. In roots of the SMIC148-A, CAT activity increased at 72 and 168h, whereas APX activity decreased at 72h and increased at 24, 12 and 168h. The Macaca showed lower root non-protein thiol group (NPSH) concentration at 200mg Al L(-1) in all evaluations, but the SMIC148-A either did not demonstrate any alterations at 24 and 72h or presented higher levels at 120h. This pattern was also observed in root ascorbic acid (AsA) concentration at 24 and 120h. The cellular redox status of these potato clones seems to be affected by Al. Therefore, oxidative stress may be an important mechanism for Al toxicity, mainly in the Al-sensitive Macaca clone.

Quercetin protects the impairment of memory and anxiogenic-like behavior in rats exposed to cadmium: Possible involvement of the acetylcholinesterase and Na+,K+-ATPase activities

The present study investigated the effects of quercetin in the impairment of memory and anxiogenic-like behavior induced by cadmium (Cd) exposure. We also investigated possible alterations in acetylcholinesterase (AChE), Na(+),K(+)-ATPase and δ-aminolevulinate dehydratase (δ-ALA-D) activities as well as in oxidative stress parameters in the CNS. Rats were exposed to Cd (2.5mg/kg) and quercetin (5, 25 or 50mg/kg) by gavage for 45days. Animals were divided into eight groups (n=10-14): saline/control, saline/Querc 5mg/kg, saline/Querc 25mg/kg, saline/Querc 50mg/kg, Cd/ethanol, Cd/Querc 5mg/kg, Cd/Querc 25mg/kg and Cd/Querc 50mg/kg. Results demonstrated that Cd impaired memory has an anxiogenic effect. Quercetin prevented these harmful effects induced by Cd. AChE activity decreased in the cerebral cortex and hippocampus and increased in the hypothalamus of Cd-exposed rats. The Na(+),K(+)-ATPase activity decreased in the cerebral cortex, hippocampus and hypothalamus of Cd-exposed rats. Quercetin prevented these effects in AChE and Na(+),K(+)-ATPase activities. Reactive oxygen species production, thiobarbituric acid reactive substance levels, protein carbonyl content and double-stranded DNA fractions increased in the cerebral cortex, hippocampus and hypothalamus of Cd-exposed rats. Quercetin totally or partially prevents these effects caused by Cd. Total thiols (T-SHs), reduced glutathione (GSH), and reductase glutathione (GR) activities decreased and glutathione S-transferase (GST) activity increased in Cd exposed rats. Co-treatment with quercetin prevented reduction in T-SH, GSH, and GR activities and the rise of GST activity. The present findings show that quercetin prevents alterations in oxidative stress parameters as well as AChE and Na(+),K(+)-ATPase activities, consequently preventing memory impairment and anxiogenic-like behavior displayed by Cd exposure. These results may contribute to a better understanding of the neuroprotective role of quercetin, emphasizing the influence of this flavonoid in the diet for human health, possibly preventing brain injury associated with Cd intoxication.

Pre-treatment with ebselen and vitamin E modulate acetylcholinesterase activity: interaction with demyelinating agents

The ethidium bromide (EB) demyelinating model was associated with vitamin E (Vit E) and ebselen (Ebs) treatment to evaluate acetylcholinesterase (AChE) activity in the striatum (ST), hippocampus (HP), cerebral cortex (CC) and erythrocytes. Rats were divided into seven groups: I—Control (saline), II—(canola); III—(Ebs), IV—(Vit E); V—(EB); VI—(EB + Ebs) and VII—(EB + Vit E). At 3 days after the EB injection, AChE activity in the CC and HC was significantly reduced in groups III, IV, V, VI and VII (p < 0.05) and in the ST it was reduced in groups III and V (p < 0.05) when compared to the control group. At 21 days after the EB injection, AChE activity in the CC was significantly reduced in groups III, IV and V, while in groups VI and VII a significant increase was observed when compared to the control group. In the HC and ST, AChE activity was significantly reduced in groups V, VI and VII when compared to the control group (p < 0.05). In the erythrocytes, at 3 days after the EB injection, AChE activity was significantly reduced in groups III, IV, V, VI and VII and at 21 days there was a significant reduction only in groups VI and VII (p < 0.05) when compared to the control group. In conclusion, this study demonstrated that Ebs and Vit E interfere with the cholinergic neurotransmission by altering AChE activity in the different brain regions and in the erythrocytes. Furthermore, treatment with Vit E and Ebs protected against the demyelination lesion caused by EB. In this context, we can suggest that ebselen and Vit E should be considered potential therapeutics and scientific tools to be investigated in brain disorders associated with demyelinating events.

Aluminum-induced oxidative stress in cucumber.

Aluminum (Al) is one of the most abundant elements of the planet and exposure to this metal can cause oxidative stress and lead to various signs of toxicity in plants. Plants are essential organisms for the environment as well as food for humans and animals. The toxic effect of aluminum is the major cause of decreased crop productivity. Thus, the objective of the present study was to analyze the effects of aluminum on the activity of antioxidant enzymes such as catalase (CAT - E.C. 1.11.1.6), superoxide dismutase (SOD - E.C.1.15.1.1) and ascorbate peroxidase (APX - E.C. 1.11.1.11), and on lipid peroxidation, electrolyte leakage percentage (ELP) and chlorophyll and protein oxidation levels in Cucumis sativus L. (cv. Aodai). Seedlings were grown at different concentrations of aluminum ranging from 1 to 2000 microM for 10 days. The increase in ELP and H(2)O(2) production observed in the seedlings may be related to the decreased efficiency of the antioxidant system at higher aluminum concentrations. The antioxidant system was unable to overcome toxicity resulting in negative effects such as lipid peroxidation, protein oxidation and a decrease in the growth of Cucumis seedlings. Aluminum toxicity triggered alterations in the antioxidant and physiological status of growing cucumber seedlings.

Rosmarinic acid prevents lipid peroxidation and increase in acetylcholinesterase activity in brain of streptozotocin-induced diabetic rats.

We investigated the efficacy of rosmarinic acid (RA) in preventing lipid peroxidation and increased activity of acetylcholinesterase (AChE) in the brain of streptozotocin‐induced diabetic rats. The animals were divided into six groups (n = 8): control, ethanol, RA 10 mg/kg, diabetic, diabetic/ethanol and diabetic/RA 10 mg/kg. After 21 days of treatment with RA, the cerebral structures (striatum, cortex and hippocampus) were removed for experimental assays. The results demonstrated that the treatment with RA (10 mg/kg) significantly reduced the level of lipid peroxidation in hippocampus (28%), cortex (38%) and striatum (47%) of diabetic rats when compared with the control. In addition, it was found that hyperglycaemia caused significant increased in the activity of AChE in hippocampus (58%), cortex (46%) and striatum (30%) in comparison with the control. On the other hand, the treatment with RA reversed this effect to the level of control after 3 weeks. In conclusion, the present findings showed that treatment with RA prevents the lipid peroxidation and consequently the increase in AChE activity in diabetic rats, demonstrating that this compound can modulate cholinergic neurotransmission and prevent damage oxidative in brain in the diabetic state. Thus, we can suggest that RA could be a promising compound in the complementary therapy in diabetes. Copyright

Effect of cadmium on growth, micronutrient concentration, and δ-aminolevulinic acid dehydratase and acid phosphatase activities in plants of Pfaffia glomerata

Plântulas de Pfaffia glomerata (Spreng.) Pedersen foram cultivadas em cinco niveis (0, 20, 40, 60 e 80 μM) de cadmio (Cd) em um sistema hidroponico durante 7 d, visando-se analisar o crescimento, as concentracoes de micronutrientes, clorofilas e carotenoides, bem como as atividades da desidratase do acido δ-aminolevulinico (ALA-D; E.C.4.2.1.24) e fosfatase acida (AP; E.C.3.1.3.2) nas plantas. A concentracao de Cd, na parte aerea e raizes, aumentou com o incremento dos niveis de Cd. A concentracao de Cd nas raizes foi, em media, 12 vezes maior do que na parte aerea. O comprimento das raizes nao foi afetado pelos tratamentos de Cd. Em contraste, a biomassa seca da parte aerea e raizes aumentou significativamente pela adicao de 20 e 40 μM Cd. Alem disso, a biomassa seca total das plantas somente diminuiu no nivel de 80 μM Cd em relacao a das plantas-controle. Por outro lado, a biomassa radicular diminuiu significativamente pela adicao de Cd a niveis superiores a 40 μM. Constatou-se resposta dependente do orgao e do micronutriente a toxicidade de Cd. As concentracoes de Zn e Cu na parte aerea e raizes nao foram alteradas pela presenca de Cd. A absorcao de Mn foi diminuida pelo estresse de Cd, porem sua translocacao nao foi alterada. Um efeito sinergistico do Cd na concentracao de Fe nas raizes foi observado nos niveis de 20 e 80 μM Cd. As atividades da ALA-D e AP foram diminuidas com o incremento dos niveis de Cd, porem a ALA-D foi mais afetada. Naquelas concentracoes de Cd, a concentracao de clorofila tambem foi diminuida. Houve uma correlacao positiva entre as concentracoes de carotenoides e clorofila. Os resultados indicam que a P. glomerata parece ter algum grau de tolerância ao Cd.

Alterations in the cholinesterase and adenosine deaminase activities and inflammation biomarker levels in patients with multiple sclerosis.

Multiple sclerosis (MS) is one of the main chronic inflammatory diseases of the CNS that cause functional disability in young adults. It has unknown etiology characterized by the infiltration of lymphocytes and macrophages into the brain. The aim of this study was to evaluate the acetylcholinesterase (AChE) activity in lymphocytes and whole blood, as well as butyrylcholinesterase (BChE) and adenosine deaminase (ADA) activities in serum. We also checked the levels of nucleotides, nucleosides, biomarkers of inflammation such as cytokines (interleukin (IL)-1, IL-6, interferon (IFN)-γ, tumor necrosis factor-alpha (TNF-α) and IL-10) and C-reactive protein (CRP) in serum from 29 patients with the relapsing-remitting form of MS (RRMS) and 29 healthy subjects as the control group. Results showed that AChE in lymphocytes and whole blood as well as BChE, and ADA activities in serum were significantly increased in RRMS patients when compared to the control group (P<0.05). In addition, we observed a decrease in ATP levels and a significant increase in the levels of ADP, AMP, adenosine and inosine in serum from RRMS patients in relation to the healthy subjects (P<0.05). Results also demonstrated an increase in the IFN-γ, TNF-α, IL-1, IL-6 and CRP (P<0.05) and a significant decrease in the IL-10 (P<0.0001) in RRMS patients when compared to control. Our results suggest that alterations in the biomarkers of inflammation and hydrolysis of nucleotides and nucleosides may contribute to the understanding of the neurological dysfunction of RRMS patients.

Photosynthetic pigments content, δ-aminolevulinic acid dehydratase and acid phosphatase activities and mineral nutrients concentration in cadmium-exposed Cucumis sativus L.

In this study, the effects of cadmium chloride (CdCl2) on plant growth, histology of roots, photosynthetic pigments content, δ-aminolevulinic acid dehydratase (ALA-D; E.C. 4.2.1.24) and acid phosphatase activities (AP; E.C. 3.1.3.2), soluble phosphorus (Pi) measurement and mineral nutrients content in cucumber seedlings (Cucumis sativus L.) were investigated. Cucumber seedlings were grown in vitro in an agar-solidified substrate containing four CdCl2 treatments (0, 100, 400, and 1000 μM) for ten days. Cd was readily absorbed by seedlings and its content was greater in the roots than in the shoot. Cd reduced shoot and root length, and fresh and dry biomass of seedlings. Inhibition of root cell elongation in Cd-treated seedlings was observed by the increase of the mean radial size of cells belonging to three zones of the root tip. The highest level of Cd reduced in a similar manner chlorophyll a, chlorophyll b and total chlorophyll contents. Increasing concentrations of Cd resulted in a linear decrease in carotenoids levels of cotyledons. Interestingly, the ALA-D activity in cotyledons was inhibited only at the highest level of Cd. Root and shoot AP activities were, respectively, activated and inhibited at all CdCl2 concentrations. Root Pi concentration was increased in all Cd treatments and it was not altered in the shoot tissues. Moreover, in general, the nutrient contents were increased in the root and decreased in the shoot. Therefore, we suggest that Cd affects negatively growth, photosynthetic pigments, ALA-D and AP activities and partition of mineral nutrients in cucumber seedlings.