Novel type of ion channel activated by Pb2+, Cd2+, and Al3+ in cultured mouse neuroblastoma cells

Summary Superfusion with Pb²⁺ induces a slow, noninactivating and reversible inward current in voltage-clamped N1E-115 neuroblastoma cells. The amplitude of this inward current increases in the range of 1–200 m Pb²⁺. Single-channel patch-clamp experiments have revealed that this inward current is mediated by discrete ion channels. Reversal potentials from linearI–V relationships are close to 0 mV for whole-cell and single-channel currents and the single-channel conductance amounts to 24 pS. The Pb²⁺-induced membrane current is not mediated by various known types of ion channels, since it is not blocked by external tetrodotoxin, tetraethylammonium,d-tubocurarine, atropine, ICS 205-930 and by internal EGTA. In Na⁺-free solutions superfusion with Pb²⁺ neither evokes a whole-cell inward current, nor single-channel openings. At –80 mV the open-time distribution of the single channels activated by 1m Pb²⁺ is dual exponential with time constants of 17 and 194 msec. When the Pb²⁺ concentration is increased from 1 to 20 m these time constants decrease to 2 and 13 msec, but the amplitude of single-channel currents remains –1.9 nA. Cd²⁺ and Al³⁺ induce inward currents and single-channel openings similar to Pb²⁺. Time constants fitted to the open-time distribution of single channels are 14 and 135 msec in the presence of 1 m Cd²⁺ and 15 and 99 msec in the presence of 50 m Al³⁺. Conversely, Cu²⁺ induces an irreversible inward current in neuroblastoma cells. Single-channel openings are undetected in the presence of Cu²⁺ and in Na⁺-free solutions Cu²⁺ is still able to induce an inward current. It is concluded that Pb²⁺, Cd²⁺ and possibly Al³⁺ activate a novel type of metal ionactivated (MIA) channel in N1E-115 cells.     

重金属Cd2+、Pb2+ 和Zn2+ 对泥鳅 DNA损伤的研究

采用单细胞凝胶电泳技术(SCGE),研究重金属Cd2 、Pb2 、Zn2 在不同暴露时间(1—35d)、单一重金属离子不同暴露浓度(0.05mg/L、0.5mg/L、5.0mg/L)或混合重金属离子(Cd2 Pb2 、Cd2 Zn2 、Pb2 Zn2 、Cd2 Pb2 Zn2 )相同浓度(0.5mg/L)条件下对泥鳅肝胰脏细胞核DNA的损伤作用。以带彗尾核DNA百分率和彗尾长度(TL)与核直径(D)比值为指标,探讨DNA损伤级别与处理浓度间的相关性。结果显示,随着处理时间的延长,带彗尾核DNA百分率和TL/D值均呈上升趋势,5.0mg/L Zn2 组28d时带彗尾核DNA百分率最高(84.85%),35d的TL/D值亦为所有组中最高(2.50);对DNA损伤作用,初期以1级损伤为主,7d后以3级损伤为主,且损伤率超过80%;Cd2 、Pb2 和Zn2 之间的联合毒性表现复杂,但总体表现为Cd2 存在时能增强Pb2 或Zn2 对DNA的损伤作用。总之,重金属Cd2 、Pb2 和Zn2 对泥鳅肝胰脏细胞核DNA损伤具有明显的浓度和时间效应,利用SCGE技术可对水环境污染导致的生物基因毒性作用进行监测。     

The effects of Ni2+, Co2+, and Mn2+ on human serum butyrylcholinesterase

The effects of Ni2+, Co2+, and Mn2+ on human serum butyrylcholinesterase (BChE, acylcholine acylhydrolase E.C. 3.1.1.8) were investigated in this study. Inhibition kinetics of BChE were studied using butyrylthiocholine (BTCh) as substrate. The "1/v" versus "1/[BTCh]" plots in the absence (control plot) and in the presence of the metal ions intersected above 1/[BTCh]-axis for all trace elements. In addition, when the concentrations of the cations were increased at 4 mM BTCh, velocities decreased and drove to zero at high concentrations of the trace elements. These results demonstrate that Ni2+, Co2+, and Mn2+ are linear mixed-type inhibitors of BChE. alphaK(i) values have been determined as 53.20 mM,152.25 mM, and 190.24 mM for Ni2+, Mn2+, and Co2+, respectively, by using nonlinear regression analysis. From the comparison of alphaK(i) values of the trace elements, it can be said that BChE has more affinty to binding Ni2+ than Co2+ and Mn2+.     

Effects of heavy metals Cd2+, Pb2+ and Zn2+ on DNA damage of loach Misgurnus anguillicaudatus

The effects of heavy metals Cd2+, Pb2+ and Zn2+ at 0.05, 0.5 and 5.0 mg/L level and their interactions at 0.5 mg/L level on DNA damage in hepatopancreas of loach Misgurnus anguillicaudatus for 1-35 days exposure were examined by single cell gel electrophoresis (SCGE). For each test group, 20 loaches with similar body size (5.17-7.99g; 11.79-13.21 cm) were selected and kept in aquaria with dechlori-nated water at (22±1)℃ and fed a commercial diet every 48 h. According to the percentage of damaged DNA with tail and its TL/D (tail length to diameter of nucleus) value, the relationship between DNA damage degree and heavy metal dose and exposure time was determined. Results showed that the percentage of damaged DNA and the TL/D value were increased with the prolonged exposure time. The highest percentage (84.85%) of damaged DNA was shown in 5.0 mg/L Zn2+ group after 28 days exposure and the biggest TL/D value (2.50) in all treated groups after 35 days exposure. During the first treated week, the damnification of DNA was mainly recognized as the first level, after that time, the third damaged level was mostly observed and the percentage of damaged DNA was beyond 80%. The joint toxic effects among Cd2+, Pb2+ or Zn2+ revealed much complexity, but it generally displayed that the presence of Cd2+ could enhance the genotoxicity of Pb2+ or Zn2+. In conclusion, the results suggested that there was a significant time-and dose-depended relationship between the heavy metal and DNA damage in hepatopancreas of loach, and SCGE could represent a useful means to evaluate the genotoxicity of environmental contamination on aquatic organisms.     

Investigation of Vetiver Grass Capability in Phytoremediation of Contaminated Soils with Heavy Metals (Pb,Cd, Mn,and Ni)

ABSTRACT

Soil contamination with heavy metals has become a worldwide concern. A sustainable technology to mitigate heavy metal contamination is extremely important. Phytoremediation is a cost-effective method, environmentally friendly, and esthetically pleasing. The aim of this study was to investigate the potential of Vetiver phytoremediation of soils contaminated with heavy metals. This research was conducted as a factorial design with four different heavy metals (lead, cadmium, manganese, and nickel) with three varying levels and also three replications for each treatment. Statistical analysis of data was performed using SPSS19 software and analysis of variance, Duncan and Pearson correlation tests. The results showed that, the highest uptake rate was related to lead metal with 282.45 mg/kg of dry soil and 83.4% uptake percentage. Then, the mean and percentage of adsorption for cadmium, nickel and manganese were 248.3 mg/kg (53.2%), 69.4 mg/kg (65.5%), and 63.29 mg/kg (61%), respectively. Lead was found to be the main component of uptake by Vetiver plant. It was found that the roots of the plant have absorbed more heavy metals than the shoots. And at the roots in total 1089.05 and on average 363.01 mg/kg and at the shoots 901.19 and on average 300.39 mg/kg, the metals used were adsorbed on three levels and four treatments. The results of analysis of variance, Duncan test and Pearson correlation showed that the effect of applied treatments on lead uptake in roots and shoots increased significantly (P ≤ 0.05) with increasing levels of treatments. The biological concentration factor was more than one, and the transfer factor was close to one. Therefore, it can be used as a phytostablization plant. The results showed that Vetiver can be considered as a refining plant due to its vegetative characteristics, cost-effectiveness and high adaptation to environmental conditions.     

Removal of Cd2+, Pb2+, and Zn2+ from contaminated water using dolomite powder

Dolomite collected from Surat Thani Province in Thailand was investigated for use as a sorbent for the removal of divalent heavy metal cations from an aqueous solution. The sorbent had a surface area of 2.46 m²/g and a pH of zero point charge (pHzpc) of 9.2. Batch sorption was used to examine the effect of the pH (pH 3–7) on the sorption capacity of Cd²⁺, Pb²⁺ and Zn²⁺, alone or together as an equimolar mixture at various concentrations. Alone, each heavy metal cation was adsorbed faster at a higher pH, where the sorption of Cd²⁺ and Pb²⁺ fitted a Langmuir isotherm, but Zn²⁺ sorption best fitted a Freundlich isotherm. Under equimolar competitive sorption, the sorption capacity of each cation was decreased by 75.8% (0.29–0.07 mM/g), 82.8% (0.53–0.09 mM/g), and 95.7% (0.84–0.04 mM/g) for Cd²⁺, Pb²⁺ and Zn²⁺, respectively, compared to that with the respective single cation. Desorption of these heavy metal cations from dolomite was low, with an average desorption level of 0.06–17.4%. Furthermore, since dolomite is readily available and rather cheap, it is potentially suitable for use as an efficient sorbent to sorb Cd²⁺ and Pb²⁺, and perhaps Zn²⁺, from contaminated water.     

Effects of Ce3+, Cd2+, and Hg2+ on activities and secondary structure of trypsin

The different effects of Ce³⁺, Cd²⁺, and Hg²⁺ on the activities and secondary structure of trypsin were studied. The results showed that trypsin activity was increased substantially by Ce³⁺ in 0.5–5 μmol/L concentration, but the activity was decreased significantly by Cd²⁺ or Hg²⁺ in 0.5–5 μmol/L concentration. The ultraviolet-visible spectrum of trypsin with 4 μmol/L Ce³⁺ treatment was the same as that of the control, but the 232-nm characteristic peak of trypsin with 4 μmol/L Cd²⁺ or Hg²⁺ treatment was blue-shifted and the peak intensity weakened. The circular dichroism (CD) spectrum of trypsin with 4 μmol/L Ce³⁺ treatment was similar to that of the control. The secondary structure of trypsin did not change with Ce³⁺ treatment. However, the CD spectrum of trypsin with 4 μmol/L Cd²⁺ or Hg²⁺ treatment was different from that of the control and Ce³⁺ treatment. The secondary structure of trypsin with Cd²⁺ or Hg²⁺ treatment changed greatly; for example, the α-helix and β-sheet contents were reduced significantly, the β-turn was enhanced greatly, and the random coil contents increased or decreased.     

Effects of Hg2+ and cell conditions on Pb2+ accumulation by Saccharomyces cerevisiae

Pb²⁺ accumulation in Saccharomyces cerevisiae changed by Hg²⁺ and cell conditions. The accumulated Pb²⁺ amounts decreased from 0.22 to 0.02?mmol Pb²⁺/g cell dry weight by the existence of Hg²⁺. But the total metals accumulation (0.42?mmol metal ions/g cell dry weight) was not changed. The order of accumulated Pb²⁺ amounts (mg Pb²⁺/g cell dry weight) according to the cell conditions at an equilibrium state was shown as the original cell (260)?>?5?times autoclaved cell for 15?min (150)?>?grinded cell after drying (100)?>?autoclaved cell for 5?min (30).     

Combined actions of Pb2+, Zn2+, and Al3+ on voltage-activated calcium channel currents

Summary 1. Whole-cell patch clamp experiments were performed on rat dorsal root ganglion (DRG) neurons to investigate the actions of various combinations of Pb²⁺, Zn²⁺, and Al³⁺ on voltage-activated calcium channel currents (VACCCs).2. Each of these metals has been shown to reduce VACCCs.3. We investigated the effects of simultaneous application of two cations in the range of their IC₅₀ values. For all possible combinations (Pb²⁺/Zu²⁺, Zn²⁺/Al³⁺, Al³⁺/Pb²⁺), independent of the order of application, we found additive actions on VACCCs.4. We observed a 75% (±9%) block of the control current when two cations were applied simultaneously. This observation is consistent with both, an action of two metals at the same site as well as independent actions at different locations of the ion channel.5. The additivity of the effects should be taken into account for questions of public health and the assessment of threshold limits in cases of environmental contamination.     

Effects of mine effluent on uptake of Co,Ni, Cu,As, Zn,Cd, Cr and Pb by aquatic macrophytes

Concentrations of Ni, Co, Cu, Pb, Zn, Cd, Cr and As were determined in aquatic sediments, water and macrophytes collected from a fluvial system, contaminated by mine effluents. Myriophyllum verticillatum collected in May below the trace element point source accumulated 169 µg/g of Ni, 860 µg/g of Co, 37 µg/g of Cu, 31 µg/g of Pb, 92 µg/g of Zn, 6.9 µg/g of Cr and 1,200 µg/g of As (concentrations in dry weight). The aquatic macrophytes Nymphaea odoratae and Pontederia cordata accumulated the investigated trace elements to a much lesser extent. The concentrations of trace elements in Myriophyllum verticillatum decreased from May to August. Correlations were found between the concentrations of total Ni, Co and Cu in the bottom sediment and in the submerged macrophytes. However, there was no correlation between the amounts of these trace elements extractable by 0.5 N HCl from the sediments and the concentrations in the macrophytes.     

铜、铅、镉、锌、汞和银离子复合污染对水螅的急性毒性效应

以水螅(Hydrasp)为例,通过单因子静态急性毒性试验方法和等毒性溶液法,分别研究Hg2 、Cu2 、Cd2 、Ag 、Zn2 和Pb2 对其单一和复合毒性效应。单一实验结果表明,它们对水螅毒性大小顺序为Hg2 >Cu2 >Cd2 >Ag >Zn2 >Pb2 。复合毒性实验表明,Zn2 与Cu2 、Hg2 、Pb2 、Ag ;Pb2 与Cu2 ;Hg2 与Ag ;Pb2 与Ag 这些组合对水螅联合急性毒性总体上表现出拮抗作用,Cd2 与Cu2 、Hg2 、Pb2 、Ag 组合总体上则是协同作用,Zn2 与Cd2 、Pb2与Hg2 、Cu2 与Hg2 ,Ag 在不同的浓度水平组合下明显表现出不同的毒性效应。     

Hg2+、Pd2+、Cd2+和Cu2+对沼泽红假单胞菌生长及净化氮、磷能力的影响

研究了在培养基中分别添加不同浓度的重金属离子Hg2+、Pd2+、Cd2+和Cu2+作为人工配制的污水对光合细菌沼泽红假单胞菌生长与净化PO4-P和NH4-N能力的影响.实验结果表明当培养基中Hg2+的浓度达到2×10-6mol@L-1,该菌生长趋势开始减慢;当增至4×10-6mol@L-1时,生长完全被抑制.Cu2+的浓度达到1×10-6mol@L-1,该菌生长趋势也开始减慢;当增至8×10-6mol@L-1时生长完全被抑制.Cd2+的浓度达4×10-5mol@L-1时,其延缓期大大延长;增至16×10-5mol@L-4时生长完全被抑制.培养基中Pd2+的浓度达到8×10-4mol@L-1时,对该菌生长并未产生影响.当培养基中重金属离子浓度未达到完全抑制其生长时,对其净化PO4-P和NH4-N能力的影响不显著.     

Multifunctional peptide‐based fluorescent chemosensor for detection of Hg2+, Cu2+ and S2− ions

A novel multifunctional fluorescent peptide sensor based on pentapeptide dansyl‐Gly‐His‐Gly‐Gly‐Trp‐COOH (D‐P5) was designed and synthesized efficiently using Fmoc solid‐phase peptide synthesis (SPPS). This fluorescent peptide sensor shows selective and sensitive responses to Hg²⁺ and Cu²⁺ among 17 metal ions and six anions studied in N‐2‐hydroxyethylpiperazine‐N‐2‐ethane sulfonic acid (HEPES) buffer solution. The peptide probe differentiates Hg²⁺ and Cu²⁺ ions by a ‘turn‐on’ response to Hg²⁺ and a ‘turn‐off’ response to Cu²⁺. Upon addition of Hg²⁺ or Cu²⁺ ions, the sensor displayed an apparent color change that was visible under an ultraviolet lamp to the naked eye. The limits of detection (LOD) of DP‐5 were 25.0 nM for Hg²⁺ and 85.0 nM for Cu²⁺; the detection limits for Cu²⁺ were much lower than the drinking water maximum contaminant levels set out by the United States Environmental Protection Agency (USEPA). It is noteworthy that both D‐P5‐Hg and D‐P5‐Cu systems were also used to detect S^2? successfully based on the formation of ternary complexes. The LODs of D‐P5‐Hg and D‐P5‐Cu systems for S^2? were 217.0 nM and 380.0 nM, respectively. Furthermore, the binding stoichiometry, binding affinity and pH sensitivity of the probe for Hg²⁺ and Cu²⁺ were investigated. This study gives new possibilities for using a short fluorescent peptide sensor for multifunctional detection, especially for anions.     

有机螯合剂在芦苇富集转运铅中的作用

近年来,随着人口的激增和工农业生产的迅猛发展,土壤和水体的重金属污染日益加重。植物提取技术作为一种新兴的修复重金属污染的治理方法,以其潜在的高效、经济及其生态协调性等优势被广泛关注。其研究主要是利用对某些重金属具有超富集能力的超积累植物或耐性植物,配合添加一定量的鳌合剂来进行修复。由于超富集植物一般生长缓慢且生物量小,因此利用对重金属有一定耐性的生长快速、生物量大的植物来进行修复更具有广阔的应用前景。芦苇作为一种多年生禾本科植物,具有生物量大、耐受性强和分布广泛的特点,已经被研究和应用到环境治理的多个领域。本文通过户外盆栽沙培试验,研究了铅胁迫下,分别在 2-10 mmol·L-1共 5 个浓度梯度的 EDTA 和柠檬酸两种鳌合剂处理下,芦苇生长对铅的响应以及对铅在植物体内的富集转运的影响。初试结果表明∶在 4mmol·L-1 Pb(NO3)2胁迫下,施加 EDTA 对芦苇的生长具有一定的抑制作用。当 EDTA 浓度为 10 mmol·L-1 时,透灌 7 d 后,其芦苇根和地上部(茎和叶)干重分别是空白试验(未加螯合剂)的 68.0% 和 72.3%,而柠檬酸对植物生物量的影响较小。同时,EDTA 和柠檬酸对铅在芦苇体内的富集和转运,都起到了促进作用。分别施加 10 mmol·L-1的 EDTA 和柠檬酸,其植物根部和地上部的铅浓度分别为空白试验的 7.4 倍、10.7 倍、2.5 倍、3.5 倍。EDTA 在促进植物体内铅的吸收和从根部到地上部的转运效率方面要比柠檬酸更为有效。尽管像 EDTA 和柠檬酸这样的鳌合剂能够在一定程度上提高植物的富集效率,但考虑到螯合剂自身对植物的毒害性、对环境的潜在威胁以及运行成本等方面因素,目前,寻找高效、廉价、可生物降解的鳌合剂以及大生物量的耐性植物成为植物提取技术的研究重点。     

Hg2+、Cd2+和Cu2+对菹草光合系统及保护酶系统的毒害作用

研究了Hg2 、Cd2 、Cu2 对高等水生沉水植物菹草光合系统及保护酶系统的毒害作用。结果表明 :3种离子均使菹草叶片叶绿体自发荧光强度、叶绿素含量、光合速率降低。Hg2 、Cd2 、Cu2 对菹草保护酶系统存在不同的影响 ,短时间低浓度条件下 ,诱导超氧化物歧化酶 (SOD )、过氧化氢酶 (CAT)、过氧化物酶(POD)活性上升 ,随着污染时间的延长、污染浓度的增加 ,酶活性下降 ,其中SOD活性上升持续时间最长 ,下降最慢。 3种离子均使菹草可溶性蛋白含量减少。Hg2 、Cu2 毒性较强 ,Cd2 毒性较弱。Hg2 、Cu2 对菹草的致死浓度为 0 .5～ 1mg/L ,Cd2 为 1～ 2 .5mg/L     

Impacts of Vehicle Emissions and Ambient Atmospheric Deposition in Nigeria on the Pb,Cd, and Ni Content of Fermented Cassava Flour Processed by Sun-Drying

A study of air pollution in Nigeria due to Pb, Cd, and Ni contamination of fermented cassava meant for flour production was carried out by AAS. Fermented cassava samples were purchased from farmers in the study area and one-third of each was oven-dried (OD) while the rest were sun-dried either along roadside (RS-SD) or under ambient atmosphere conditions (AA-SD). Mean concentrations (μg/g) for OD samples in year 2008 were: Pb (0.17 ± 0.04); Cd (0.04 ± 0.01); and Ni (0.27 ± 0.05) while RS-SD samples gave Pb (0.24 ± 0.08), Cd (0.04 ± 0.01), and Ni (0.48 ± 0.17). Mean values (μg/g) in 2009 for OD, AA-SD, and RS-SD, respectively, were Pb (0.03 ± 0.01, 0.05 ± 0.02, 0.15 ± 0.08), Cd (0.18 ± 0.01, 0.04 ± 0.02, 0.05 ± 0.02), and Ni (0.09 ± 0.06, 0.21 ± 0.10, 0.57 ± 0.12). Mean concentrations in sun-dried samples were greater (p < .01), while RS-SD samples were 185% in Pb, 53% in Cd, and 176% in Ni greater (p < .01) than AA-SD samples. With an estimated country-wide annual Pb emissions from petrol and diesel ranging from 616,241 to 968,086 kg, and dust, considered the major source of Cd and Ni, and other extraneous factors including metals adsorbed on pavement surfaces, sun-drying of wet foodstuff on the bare surface of roadside pavements could lead to high levels of Pb, Cd, and Ni in such food compared to drying under factory conditions or oven-drying.     

Accumulation of Ag, Cd, Co, Cu, Hg, Ni and Pb in Pavlova viridis Tseng (Haptophyceae)

The flagellate alga Pavlova viridis Tseng was investigated in the laboratory for accumulation of the heavy metals, silver, cadmium, cobalt, copper, mercury, nickel and lead. The cultures were grown in an artificial seawater medium mixed with the individual metals at different concentrations. Based on data from the controls, the baseline metal concentrations in P. viridis were shown to be in an order of Cu > Pb > Co > Cd > Ni > Ag > Hg. In the experimental groups, the seven metals displayed different isotherm equilibrium patterns and the metal uptake capacity of the alga was Ni > Pb > Co > Hg > Cu > Cd > Ag at equilibrium. When assessed using the bioconcentration factors, metal accumulation by P. viridis was demonstrated to be the most efficient at a concentration of 0.001 mg L-1 for Ag, Cd and Co, and at 0.01 mg L-1 for Cu, Hg, Ni and Pb. This study suggests that P. viridis can be a source of mineral supplements in mariculture. The alga is not, however, recognized as an effective agent for removing heavy metals from wastewater. This revised version was published online in June 2006 with corrections to the Cover Date.     

Interactions between toxic (As, Cd, Hg and Pb) and nutritional essential (Ca, Co, Cr, Cu, Fe, Mn, Mo, Ni, Se, Zn) elements in the tissues of cattle from NW Spain

Since the toxicity of one metal or metalloid can be dramatically modulated by the interaction with other toxic or essential metals, studies addressing the chemical interactions between trace elements are increasingly important. In this study correlations between the main toxic (As, Cd, Hg and Pb) and nutritional essential (Ca, Co, Cr, Cu, Fe, Mn, Mo, Ni, Se, Zn) elements were evaluated in the tissues (liver, kidney and muscle) of 120 cattle from NW Spain, using Spearman rank correlation analysis based on analytical data obtained by ICP-AES. Although accumulation of toxic elements in cattle in this study is very low and trace essential metals are generally within the adequate ranges, there were significant associations between toxic and essential metals. Cd was positively correlated with most of the essential metals in the kidney, and with Ca, Co and Zn in the liver. Pb was significantly correlated with Co and Cu in the liver. A large number of significant associations between essential metals were found in the different tissues, these correlations being very strong between Ca, Cu, Fe, Mn, Mo and Zn in the kidney. Co was moderately correlated with most of the essential metals in the liver. In general, interactions between trace elements in this study were similar to those found in polluted areas or in experimental studies in animals receiving diets containing high levels of toxic metals or inadequate levels of nutritional essential elements. These interactions probably indicate that mineral balance in the body is regulated by important homeostatic mechanisms in which toxic elements compete with the essential metals, even at low levels of metal exposure. The knowledge of these correlations may be essential to understand the kinetic interactions of metals and their implications in the trace metal metabolism.