Peptidic models for the binding of Pb(II), Bi(III) and Cd(II) to mononuclear thiolate binding sites

Herein, we evaluate the binding of Pb(II) and Bi(III) to cysteine-substituted versions of the TRI peptides [AcG-(LKALEEK)₄G-NH₂] which have previously been shown to bind Hg(II) and Cd(II) in unusual geometries as compared with small-molecule thiol ligands in aqueous solutions. Studies of Pb(II) and Bi(III) with the peptides give rise to complexes consistent with the metal ions bound to three sulfur atoms with M–S distances of 2.63 and 2.54 Å, respectively. Competition experiments between the metal ions Pb(II), Cd(II), Hg(II) and Bi(III) for the peptides show that Hg(II) has the highest affinity, owing to the initial formation of the extremely strong HgS₂ bond. Cd(II) and Pb(II) have comparable binding affinities at pH > 8, while Bi(III) displays the weakest affinity, following the model, M(II) + (TRI LXC)₃ ^3? → M(II)(TRI LXC)₃ ^?. While the relevant equilibria for Hg(II) binding to the TRI peptides corresponds to a strong first step forming Hg(TRI LXC)₂(HTRI LXC), followed by a single deprotonation to give Hg(TRI LXC)₃ ^?, the binding of Cd(II) and Pb(II) is consistent with initial formation of M(II)(TRI LXC)(HTRI LXC)₂ ⁺ at pH < 5 followed by a two-proton dissociation step (pK _a2) yielding M(II)(TRI LXC)₃ ^?. Pb(II)(TRI LXC)(HTRI LXC)₂ ⁺ converts to Pb(II)(TRI LXC)₃ ^? at slightly lower pH values than the corresponding Cd(II)–peptide complexes. In addition, Pb(II) displays a lower pK _a of binding to the “d”-substituted peptide, (TRI L12C, pK _a2 = 12.0) compared with the “a”-substituted peptide, (TRI L16C, pK _a2 = 12.6), the reverse of the order seen for Hg(II) and Cd(II). Pb(II) also showed a stronger binding affinity for TRI L12C (K _bind = 3.2 × 10⁷ M^?1) compared with that with TRI L16C (K _bind = 1.2 × 10⁷ M^?1) at pH > 8.     

The clastogenic effects of chronic exposure to particulate and soluble Cr(VI) in human lung cells

Hexavalent chromium (Cr(VI)) is a well-designated human lung carcinogen, with solubility playing an important role in its carcinogenic potential. Although it is known that particulate or water-insoluble Cr(VI) compounds are more potent than the soluble species of this metal, the mechanisms of action are not fully elucidated. In this study, we investigated the hypothesis that the difference in potency between particulate and soluble Cr(VI) is due to more chronic exposures with particulate chromate because it can deposit and persist in the lungs while soluble chromate is rapidly cleared. Chronic exposure to both insoluble lead chromate and soluble sodium chromate induced a concentration and time-dependent increase in intracellular Cr ion concentrations in cultured human lung fibroblasts. Intracellular Pb levels after chronic exposure to lead chromate increased in a concentration-dependent manner but did not increase with longer exposure times up to 72 h. We also investigated the effects of chronic exposure to Cr(VI) on clastogenicity and found that chronic exposure to lead chromate induces persistent or increasing chromosome damage. Specifically, exposure to 0.5 μg/cm² lead chromate for 24, 48 and 72 h induced 23, 23 and 27% damaged metaphases, respectively. Contrary to lead chromate, the amount of chromosome damage after chronic exposure to sodium chromate decreased with time. For example, cells exposed to 1 μM sodium chromate for 24, 48 and 72 h induced 23, 13 and 17% damaged metaphases, respectively. Our data suggest a possible mechanism for the observed potency difference between soluble and insoluble Cr(VI) compounds is that chronic exposure to particulate Cr(VI) induces persistent chromosome damage and chromosome instability while chromosome damage is repaired with chronic exposure to soluble Cr(VI).     

An assessment of time-dependent effects of lead exposure in algerian mice (Mus spretus) using different methodological approaches

Time-dependent effects of lead (Pb) toxicity were studied in Algerian mice (Mus spretus) treated with Pb acetate via drinking water (1 g Pb acetate/L) for different periods of exposure (15, 45, and 90 d). End points included the determination of hepatic Pb concentration and the assessment of some morphophysiological, biochemical and cytogenetical parameters. A control group receiving distilled water was also monitored for comparative purposes. Hepatic Pb accumulation increased with the time of exposure and was significantly higher in treated mice when compared to controls. In association with significant body mass loss in Pb-exposed mice, for 15 and 45 d, a significant increase in the relative spleen mass was observed after 45 d of intoxication. Pb-exposed mice also showed significant decreases in red blood cells, hematocrit, and mean corpuscular hemoglobin. On the contrary, changes in plasma transferases (aspartate aminotransferase and alanine aminotransferase) and hepatic superoxide dismutase activities did not reach statistical significance. A significant increase in the frequency of micronucleated polychromatic bone marrow erythrocytes was also found in the 90-d-exposed mice, compared to nontreated mice and the other exposed groups. Exposure to Pb acetate resulted also in a slight time-dependent decrease of the polychromatic-normochromatic ratio. These results support the concept that a long-term chronic exposure to Pb induced alterations upon some morphophysiological and genetic paramaters in Algerian mice.     

锌对铅染毒新生大鼠成骨细胞增殖分化的影响

目的:探讨铅锌联合染毒对乳鼠颅骨成骨细胞增殖分化的影响。方法:分离并培养原代成骨细胞,加入不同浓度铅、锌培养48h,检测其对成骨细胞增殖的作用;用碱性磷酸酶试剂盒检测ALP活力。结果:在染铅48h后,当铅浓度≥10μmol/L时,细胞增殖功能下降(P<0.05);加锌干预48h后,铅+锌组细胞增殖功能均高于各自单独染铅组,其中铅(1μmol/L、10μmol/L)+锌(50μmol/L)组、铅(10)+锌(100)组与对照组间的差异具有统计学意义(P<0.05)。铅干预48h后,100μmol/L铅组的ALP活力显著下(P<0.05),给予锌干预的铅锌联合染毒组,各组ALP活力均有增加,其中铅(1μmol/L、10μmol/L)+锌(50μmol/L)组ALP活力均高于对照组,而铅(100μmol/L)+锌(50μmol/L)组ALP活力低于对照组,差异均有统计学意义(P<0.05)。结论:铅对成骨细胞有毒性作用,影响其增殖和分化功能;50μmol/L锌在一定程度上可以拮抗铅对成骨细胞增殖和分化功能的损伤,且对ALP活力的作用更显著,为铅中毒骨病的防治提供一定的科学依据。     

Detection of Pseudo-nitzschia (Bacillariophyceae) species and amnesic shellfish toxins in Scottish coastal waters using oligonucleotide probes and the Jellet Rapid Test™

Species specific LSU rRNA targeted fluorescent oligonucleotide probes, designed by researchers at the Monterey Bay Aquarium Research Institute (USA) for a limited range of Pseudo-nitzschia species, were applied to unialgal cultures and Scottish field samples, to investigate possible applications in Scottish phytoplankton monitoring programmes to detect potential amnesic shellfish poisoning (ASP) toxin producing species. The existing available probe for Pseudo-nitzschia australis gave good results, positively labelling cells from cultures and field samples. However, application of the P. pungens, P. delicatissima and P. fraudulenta probes gave poor results, with little or no fluorescence label observed in field samples, while transmission electron microscopy (TEM) showed these species to be present. Comparison of the same region of the LSU sequence from cultures of P. delicatissima, isolated from Scottish waters, with the probe designed for detection of P. delicatissima isolated from Monterey Bay revealed the presence of a single base difference between the two sequences, which may have prevented the probe from hybridising to Scottish isolates and cells from field samples. In an attempt to assess the potential ASP toxin production by field populations of Pseudo-nitzschia a rapid immunodiagnostic test (the Jellet Rapid Test, JRT) for ASP toxins was examined. Results indicate that additional development of molecular probes for the detection of a range of Pseudo-nitzschia species detected in Scottish coastal waters and the use of JRT for toxin detection could conceivably provide an effective tool for broad-scale mapping of toxin events and management of coastal zone activities.     

Biosorption of lead, cadmium and mercury by immobilized Microcystis aeruginosa in a column

Immobilized Microcystis aeruginosa in a flow-through sorption column was evaluated for the potential to remove Pb²⁺, Cd²⁺ and Hg²⁺ from aqueous solutions. M. aeruginosa showed high affinity for the three heavy metals with removal efficiency of 90% for Cd²⁺ and Hg²⁺, and 80% for Pb²⁺ at saturation conditions. Competitive sorption experiments conducted in paired and ternary systems indicated that Pb²⁺ was sequestered preferentially over Cd²⁺ and Hg²⁺. The presence of Cd²⁺ interfered only slightly with the uptake of Hg²⁺, as Pb²⁺ and Hg²⁺ did with Cd²⁺. In contrast, Hg²⁺ sorption was affected by Pb²⁺ to a great extent. Desorption with 1 M HCl was completed within 25 min with high efficiency and effectiveness for the three metals. The results of this study indicate that M. aeruginosa is to be a potential biosorbent material except when Hg²⁺ and Pb²⁺ are in the same contaminated solution.     

长江江豚锌、铜、铅、镉和砷的摄入与累积

根据室内饲养的3头长江江豚(Neophocaena phocaenoides asiaeorientalis)食物鲫(Carassius auratus)中锌(Zn)、铜 (Cu)、铅(Pb)、镉(Cd)、砷(As)的浓度值和饲养记录,推算出了饲养条件下江豚这些微量元素每日及每周估计摄入 量的范围。必需元素的需求方面与世界卫生组织/联合国粮农组织(WHO/FAO)提出的人暂定每周耐受摄人量 (PTWI)相类似,可是毒性较强的元素Cd、As、Pb的摄入量大大高于人体的PTWI。对湖北天鹅洲故道收集到的一头 江豚的10种组织、器官的研究发现,摄入后的Zn、Cu、Pb、Cd、As显示出有组织、器官选择性累积的倾向。这可能主 要与鲸类组织、器官对相应的元素有特殊的要求相关。本研究的结果还显示出有必要建立饲养鲸类动物食物安全 管理规范以控制食源污染。更深入地研究各组织、器官中微量元素与生命机能的关系,应该是鲸类动物微量元素 的营养化学和生态毒理学发展的重要方向之一。     

Paralytic shellfish toxin profiles of different geographic populations of Gymnodinium catenatum (Dinophyceae) in Korean coastal waters

Paralytic shellfish poisoning toxin profiles of dinoflagellate cultures of Gymnodinium catenatum Graham from the Yellow and South Seas in Korea were investigated by high performance liquid chromatography fluorometric detection. Strains from the Yellow Sea had predominantly carbamate toxins, while strains from Sujeongri and Chindong in the South Sea contained the N‐sulfocarbamoyl toxins, Cl,2, as major components including the presence of GTX5 and dcSTX in some strains. Toxin profiles from St. Deukryang Bay strains (South Sea) showed both characteristics of those in the South Sea and those in the Yellow Sea. Thirty strains could be divided into three groups based on cluster analysis of toxin compositions. Group I (Yellow Sea strains) was distinguished from Group II (Sujeongri and Chindong strains) by the absence of GTX5 and dcSTX. Group III comprised Deukryang Bay strains. In conclusion, the Yellow Sea and the South Sea were found to have different dinoflagellate populations with different toxin compositions.