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To explore the impact of sulfur (S) fertilization on copper (Cu) metabolism in grazing Tibetan sheep, fertilizing and grazing experiments were conducted in the Southwest Grand Grassland, China. The tested pasture was randomly divided into four parts by the fencing device (2 hm²/part). The pasture received ammonium sulfate [(NH4)2SO4] treatment in summer in 2020 (0, 300, 400, and 500 kg/hm2 for part I, part II, part III, and part IV, respectively). 24 Tibetan sheep (one year old) were randomly divided into four groups, and assigned to the selected pastures (part I, part II, part III, and part IV for group I, group II, group III, and group IV, respectively). The grazing experiments lasted for 90 days. The results showed that the contents of nitrogen (N) and S in fertilized forage were markedly higher than those in unfertilized grass (P<0.01), but there was no significant difference among fertilized forages. The contents of Cu in blood and liver in the Tibetan sheep from fertilized pasture were extremely lower than those in animals from unfertilized pasture (P<0.01). The contents of Hb and PCV in the tested sheep from fertilized pasture were extremely higher than those in sheep from unfertilized pasture. The activities of serum SOD, GSH-Px, T-AOC, Cp, and CAT in sheep from fertilized pasture were extremely lower than those in sheep from no fertilized pasture (P<0.01), but there were no obvious differences among the Tibetan sheep from fertilized pasture (P>0.05). The contents of MDA in the Tibetan sheep from fertilized pasture were significantly higher than those in animals from no fertilized pasture (P<0.01). There was no obvious difference among the Tibetan sheep from fertilized pasture (P>0.05). Therefore, (NH4)2SO4 fertilization not only greatly influenced the contents of S and N in forage, but also extremely decreased Cu contents of animal tissues in the Tibetan sheep from fertilized ranges. [ABSTRACT FROM AUTHOR]

Strain S30A2 T , isolated from the acid mine drainage sediment of Mengzi Copper Mine, Yunnan, is proposed to represent a novel species of the sulphur-oxidizing genus Acidithiobacillus . Cells were Gram-stain-negative, non-endospore forming, highly motile with one or two monopolar flagella and rod-shaped. The strain was mesophilic, growing at 30-50 °C (optimum, 38 °C), acidophilic, growing at pH 2.0-4.5 (optimum, pH 2.5), and tolerant of 0-4 % (w/v; 684 mol l -1 ) NaCl. The 16S rRNA gene-based sequence analysis showed that strain S30A2 T belongs to the genus Acidithiobacillus and shows the largest similarity of 96.6 % to the type strain Acidithiobacillus caldus KU T . The genomic DNA G+C content of strain S30A2 T was 59.25 mol%. The average nucleotide identity ANIb and ANIm values between strain S30A2 T and A. caldus KU T were 70.95 and 89.78 %, respectively and the digital DNA-DNA hybridization value was 24.9 %. Strain S30A2 T was strictly aerobic and could utilize elementary sulphur and tetrathionate to support chemolithotrophic growth. The major cellular fatty acid of S30A2 T was C 19 : 1 ω7 c . The respiratory quinones were ubiquinone-8 and ubiquinone-7. Based upon its phylogenetic, genetic, phenotypic, physiologic and chemotaxonomic characteristics, strain S30A2 T is considered to represent a novel species of the genus Acidithiobacillus , for which the name Acidithiobacillus acidisediminis sp. nov. is proposed. The type strain is S30A2 T (=CGMCC 1.17059 T =KCTC 72580 T ).

Root deformation is a common challenge in container seedlings especially for vigorous-rooted tree species. The issue is significant for Chinese cork oak ( Quercus variabilis Blume.) which is grown in cost-effective polybags. In this study we examined how copper root pruning and radicle pruning (1/3 or 2/3 of radicle length) individually and together (1/3 radicle pruning in combination with copper root pruning) affected root architecture, growth and nutrient storage of nursery seedlings, and survival, growth performance and nutrient status during the first outplanting season. Copper root pruning failed to reduce root spiraling, and decreased total root length and surface area by decreasing root length and surface area of roots with diameter <5 mm. In contrast, radicle pruning decreased root deformation and induced taproot formation. However, intensity of radicle pruning impacted seedling size at planting and 2/3 radicle pruning decreased height and stem dry mass relative to the control. Root effects of combination (radicle plus copper root pruning) treatment on root development were not superior to 1/3 radicle pruning. Combination treatment decreased roots with diameter <2 mm. Overall 1/3 radicle pruning has the best potential to modify root development for vigorous-rooted Chinese cork oak seedlings grown in polybags. [ABSTRACT FROM AUTHOR]

This study attempts to speculate on the future of copper metabolism in China based on dynamic substance flow analysis. Based on tremendous growth of copper consumption over the past 63 years, China will depict a substantially increasing trend of copper in-use stocks for the next 30 years. The highest peak will be possibly achieved in 2050, with the maximum ranging between 163 Mt and 171 Mt. After that, total stocks are expected to slowly decline 147–154 Mt by the year 2080. Owing to the increasing demand of in-use stocks, China will continue to have a profound impact on global copper consumption with its high import dependence until around 2020, and the peak demand for imported copper are expected to approach 5.5 Mt/year. Thereafter, old scrap generated by domestic society will occupy an increasingly important role in copper supply. In around 2060, approximately 80% of copper resources could come from domestic recycling of old scrap, implying a major shift from primary production to secondary production. With regard to the effect of lifetime distribution uncertainties in different end-use sectors of copper stocks on the predict results, uncertainty evaluation was performed and found the model was relatively robust to these changes. [ABSTRACT FROM AUTHOR]

Objectives: Wilson disease (WD) is a rare autosomal recessive disease caused by an ATP7B gene mutation. Liver cirrhosis is an important issue that affects the clinical management and prognosis of WD patients. Blood routine examination is a potential biomarker for predicting the occurrence of liver cirrhosis in WD. We aim to construct a predictive model for the occurrence of liver cirrhosis using general clinical information, blood routine examination, urine copper, and serum ceruloplasmin through a machine learning approach., Methods: Case-control study of WD patients admitted to West China Fourth Hospital between 2005 and 2020. Patients with a score of at least four in scoring system of WD were enrolled. A machine learning model was constructed by EmpowerStats software according to the general clinical data, blood routine examination, 24 h urinary copper, and serum ceruloplasmin., Results: This study analyzed 346 WD patients, of which 246 were without liver cirrhosis. And we found platelet large cell count (P-LCC), red cell distribution width CV (RDW-CV), serum ceruloplasmin, age at diagnosis, and mean corpuscular volume (MCV) were the top five important predictors. Moreover, the model was of high accuracy, with an area under the receiver operating characteristic curve of 0.9998 in the training set and 0.7873 in the testing set., Conclusions: In conclusion, the predictive model for predicting liver cirrhosis in WD, constructed by machine learning, had a higher accuracy. And the most important indices in the predictive model were P-LCC, RDW-CV, serum ceruloplasmin, age at diagnosis, and MCV., (Copyright © 2022 The Author(s). Published by Wolters Kluwer Health, Inc.)

Wilson disease (WD) is a rare and treatable genetic disease that is found worldwide. The latest progress and author's experience in this field are described, and some notable clinical issues concerning this disease in China are discussed. Many studies have shown that in recent years, the number and age range of WD cases have increased, and more and more liver - type and atypical cases have been identi-fied ; WD is difficult to diagnose, and misdiagnosed cases are commonly seen. Heightened alertness is the key to the prevention of misdiagno-sis, and laboratory testing is an indispensible basis for diagnosis of this disease. Kayser - Fleischer ring, ceruloplasmin, urinary copper, he-patic copper, and genetic testing are of great significance for the diagnosis of WD. Hepatic copper has the highest specificity and sensitivity for WD, reaching 95%, and genetic testing has a specificity of 100% and a sensitivity of 60% - 85% for WD. Patients suspected of WD should undergo systemic copper metabolic examination. Diagnostic criteria should be strictly followed, and WD can be diagnosed if the total score is not lower than 4 ; diagnosis cannot be based on a single test. Once the diagnosis is confirmed, therapy is started regardless of the se-verity of disease. Severe cases are not encouraged to discontinue therapy due to the side effects of drugs. Fulminating cases should receive rapid decopper treatment as soon as possible. Liver transplantation may be considered for patients with acute or chronic liver failure unre-sponsive to medical therapy. Overall, patients who receive adequate care for WD have a good long - term prognosis. We need to recognize the new characteristics of this disease and improve the diagnosis and treatment of WD in China. [ABSTRACT FROM AUTHOR]

The mutations in modifier genes may contribute to some inherited diseases including Wilson disease (WD). This study was designed to identify potential modifier genes that contribute to WD. A total of 10 WD patients with single or no heterozygous ATP7B mutations were recruited for whole-exome sequencing (WES). Five hundred and thirteen candidate genes, of which the genetic variants present in at least two patients, were identified. In order to clarify which proteins might be involved in copper transfer or metabolism processes, the isobaric tags for relative and absolute quantitation (iTRAQ) was performed to identify the differentially expressed proteins between normal and CuSO4-treated cell lines. Thirteen genes/proteins were identified by both WES and iTRAQ, indicating that disease-causing variants of these genes may actually contribute to the aberrant copper ion accumulation. Additionally, the c.86C > T (p.S29L) mutation in the SLC31A2 gene (coding CTR2) has a relative higher frequency in our cohort of WD patients (6/191) than reported (0.0024 in gnomAD database) in our healthy donors (0/109), and CTR2S29L leads to increased intracellular Cu concentration and Cu-induced apoptosis in cultured cell lines. In conclusion, the WES and iTRAQ approaches successfully identified several disease-causing variants in potential modifier genes that may be involved in the WD phenotype., (© The Author(s) 2022. Published by Oxford University Press.)

High fat consumption leads to reactive oxygen species (ROS) which is associated with age-progressive neurological disorders. Cu/Zn superoxide dismutase (SOD1) is a critical enzyme against ROS. However, the relationship between SOD1 and the high-fat-induced ROS and neurodegeneration is poorly known. Here we showed that, upon treatment with a saturated fatty acid palmitic acid (PA), the SOD1 activity was decreased in mouse neuronal HT-22 cell line accompanied by elevation of ROS, but not in mouse microglial BV-2 cell line. We further showed that PA decreased the levels of copper chaperone for SOD1 (CCS) in HT-22 cells, which promoted the nuclear import of SOD1 and decreased its activity. We demonstrated that the reduction of CCS is involved in the PA-induced decrease of SOD1 activity and elevation of ROS. In addition, compared with the adult mice fed with a standard diet, the high-fat-diet adult mice presented an increase of plasma free fatty acids, reduction of hippocampal SOD1 activity and CCS, mitochondrial degeneration and long-term memory decline. Taken together, our findings suggest that the high-fat-induced lower CCS level is essential for SOD1 suppression which may be associated with neurodegeneration and cognitive decline., (Copyright © 2021 Elsevier Inc. All rights reserved.)

Background: Acute coronary syndrome (ACS) is a cardiac emergency with high mortality. Exposure to high copper (Cu) concentration has been linked to ACS. However, whether DNA methylation contributes to the association between Cu and ACS is unclear., Methods: We measured methylation level at > 485,000 cytosine-phosphoguanine sites (CpGs) of blood leukocytes using Human Methylation 450 Bead Chip and conducted a genome-wide meta-analysis of plasma Cu in a total of 1243 Chinese individuals. For plasma Cu-related CpGs, we evaluated their associations with the expression of nearby genes as well as major cardiovascular risk factors. Furthermore, we examined their longitudinal associations with incident ACS in the nested case-control study., Results: We identified four novel Cu-associated CpGs (cg20995564, cg18608055, cg26470501 and cg05825244) within a 5% false discovery rate (FDR). DNA methylation level of cg18608055, cg26470501, and cg05825244 also showed significant correlations with expressions of SBNO2, BCL3, and EBF4 gene, respectively. Higher DNA methylation level at cg05825244 locus was associated with lower high-density lipoprotein cholesterol level and higher C-reactive protein level. Furthermore, we demonstrated that higher cg05825244 methylation level was associated with increased risk of ACS (odds ratio [OR], 1.23; 95% CI 1.02-1.48; P = 0.03)., Conclusions: We identified novel DNA methylation alterations associated with plasma Cu in Chinese populations and linked these loci to risk of ACS, providing new insights into the regulation of gene expression by Cu-related DNA methylation and suggesting a role for DNA methylation in the association between copper and ACS.

Iron (Fe) oxides are intimately coupled with phosphorus and closely associated with the bioavailability of potential toxic elements (PTEs) in soil. Thus, Fe oxides may influence the stabilization of PTEs in contaminated soils amended by phosphorus. To evaluate the effects of hematite (HMT) on the stabilization of PTEs, 1-5% (by weight) of HMT was added into a contaminated red soil amended with hydroxyapatite (HAP) to simulate naturally occurring Fe oxides. The stabilization efficiencies of soil copper (Cu) and cadmium (Cd) amended with HAP in soils with low, moderate, and high content of HMT were assessed after a 60-day incubation. HAP treated the soil with high rate HMT decreased the CaCl 2 -extractable and acid-soluble fractions of Cu and Cd than that of HAP alone. In particular, CaCl 2 -extactable Cu and Cd in the soil with 5% HMT amended by HAP were 91-95% and 41-68% lower than those amended with only HAP. High content of HMT in soil could decrease the concentration of labile phosphorus in the presence of HAP, but it did not increase the concentration of NaOH-extractable inorganic phosphorus (the fraction bound to Fe oxides). The concentrations of free and crystalline Fe oxides were significantly increased by adding high dosages of HMT with or without HAP. High content of HMT in soil amended by HAP reduced metal phytotoxicity and uptake by wheat shoots than the soil containing HAP without HMT. The results indicate that HMT can promote Cu and Cd stabilization while decrease labile phosphorus in red soil amended with HAP, suggesting that phosphorus-based amendments combined with Fe oxides can be used to stabilize PTEs in contaminated red soils., Competing Interests: Declaration of competing interest The authors declare no conflict of interest., (Copyright © 2020 Elsevier Inc. All rights reserved.)

The present study investigated the biochemical toxicity and potential detoxification mechanisms in earthworms Eisenia fetida exposed to sulfamethazine (SMZ) (7.5, 15 and 30 mg kg -1 ) either alone or in combination with Copper (Cu) (100 mg kg -1 ) in soil. The results showed that increasing concentrations of SMZ in soil activated superoxide dismutase, catalase and glutathione peroxidase isozymes, suggesting reactive oxygen species (ROS) burst in earthworms. Treatment with SMZ and Cu separately or in combination caused protein oxidation and damage, elevating the synthesis of ubiquitin, the 20S proteasome, cytochrome P450 (CYP450), and heat shock protein 70 (HSP70). Such treatments also induced the activities of proteases, endoproteinase (EP) and glutathione S-transferases (GSTs). The results suggested that the ubiquitin-20S proteasome, proteases, EP and HSP70 were involved in degradation or remediation of oxidatively damaged proteins. Elevated levels of CYP450 and GSTs also participated in the detoxification of the earthworms.

Objective: To determine the relationship between ATP7B mutations and diabetes in Wilson disease (WD)., Research Design and Methods: A total of 21 exons and exon-intron boundaries of ATP7B were identified by Sanger sequencing., Results: Two novel compound heterozygous mutations (c.525 dupA/ Val176Serfs*28 and c.2930 C>T/ p.Thr977Met) were detected in ATP7B . After d-penicillamine (D-PCA) therapy, serum aminotransferase and ceruloplasmin levels in this patient were normalized and levels of HbA 1c decreased. However, when the patient ceased to use D-PCA due to an itchy skin, serum levels of fasting blood glucose increased. Dimercaptosuccinic acid capsules were prescribed and memory recovered to some extent, which was accompanied by decreased insulin dosage for glucose control by 5 units., Conclusions: This is the first report of diabetes caused by WD., (© 2020 by the American Diabetes Association.)

Water samples were collected from five phases of treatment in a municipal sewage reclaimed water plant and the DOM was characterized. Results indicated that the components and properties of DOM varied notably with sequential treatments, such as the fluorescence intensity, the molecular weight and the total acidity. Meanwhile the accumulation of Cu in Daphnia magna was analyzed following exposure to samples spiked with 50 µg/L copper, which were decreased by the presence of DOM in those water samples. Furthermore, this study found significant associations between fluorescence intensity and Cu accumulation (r 2  = 0.778, p < 0.05), while increased total acidity was found to enhance the unit total organic carbon-Cu accumulation (r 2  = 0.979, p < 0.01). This study provides useful information on the safety and effective management of reclaimed water as a potential water resource.

We assessed the efficacy of Ca-bentonite (CB) alone and combined with Ca-hydroxide (CH), tobacco biochar (TB), and zeolite (ZL) aiming to immobilize Cu and Pb and decrease their bioavailability and uptake by pak choi followed by maize in a mining contaminated soil. The CB alone was able to decrease the availability and uptake of Cu and Pb by pak choi and maize. The mono- and multi-combination of CH, TB, and ZL with CB showed contradictory impact on the availability and uptake of Cu and Pb as compared to the mono-application of CB. The combination of CB with ZL and CH + ZL reduced the uptake of Pb by pak choi and maize, while the combination of CB with TB and ZL reduced the uptake of Cu by pak choi and maize as compared to the mono-application of CB. The co-application of CB with CH increased the phytoextraction of Cu by maize and Pb by pak choi shoots as compared to the mono-application of CB. We conclude that modified clays such as CB alone or combined with ZL, TB, and/or CH might be suitable candidates for phytomanagement of Cu and Pb contaminated soils., (Published by Elsevier B.V.)

With the population growth, urbanization and industrialization, China has become a hotspot of atmospheric deposition nitrogen (ADN), which is a threat to ecosystem and food safety. However, the impacts of increased ADN on rice growth and grain metal content are little studied. Based on previous long-term ADN studies, greenhouse experiment was conducted with four simulated ADN rates of 0, 30, 60 and 90 kg N ha -1  yr -1 (CK, N 1 , N 2 and N 3 as δ 15 N, respectively) to assess rice growth and metal uptake in a red soil ecosystem of southeast China during 2016-2017. Results showed that simulated ADN could promote rice growth and increase yields by 15.68-24.41% (except N 2 ) and accumulations of cadmium (Cd) or copper (Cu) in organs. However, there was no linear relationship between ADN rate and rice growth or Cd or Cu uptake. The 15 N-ADN was mainly accumulated in roots (21.31-67.86%) and grains (25.26-49.35%), while Cd and Cu were primarily accumulated in roots (78.86-93.44% and 90.00-96.24%, respectively). 15 N-ADN and Cd accumulations in roots were significantly different between the two growing seasons (p < 0.05), implying the accumulative effects of ADN. Data also indicated the synergetic effect between accumulations of 15 N-ADN and soil Cd and Cu. This study demonstrated that increasing ADN could potentially enhance Cd or Cu uptake in rice grain and threaten rice grain safety. However, related nonlinear mechanism is still needed to be discussed between increasing ADN and rice response in the future., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Long term mining activities can cause significant metal pollution in the environment, thereby showing potential risk to the paddy field. Elucidating the interfacial processes of trace metals from contaminated paddy soil to rice within the rhizosphere can provide important information on metal biogeochemistry and food safety. The current study aims to explore the spatial distribution and molecular speciation of Cu from rhizosphere to rice plant in a mining-impacted paddy soil, and reveal the possible uptake mechanisms. X-ray absorption near edge structure (XANES) analysis indicated that Cu was primarily associated with iron oxide and sulfide in soil with a minor proportion of organic complexed species. In the rice samples, Cu showed much higher concentrations in the roots than the shoots, as most Cu was sequestered in the root surface and epidermis (primarily in the form of C/N ligands bound Cu species), rather than root xylem, as identified by micro X-ray fluorescence (μ-XRF) imaging coupling with μ-XANES. By contrast, in the root xylem, thiol-S bound Cu(I) complex was observed, representing the reduced product of Cu(II) by thiol-S ligands in rice root. The absorbed Cu was probably transported from the root to the aerial part as C/N ligand bound Cu complex such as Cu-histidine like species, which was observed in the root xylem. The large retention capacity and reduction of Cu(II) in rice root alleviated Cu toxicity to rice, which was beneficial for food safety (e.g., lower concentration of Cu in rice grains). These findings showed for the first time that the uptake mechanisms by rice from field contaminated sites, which shed light on Cu detoxification process and potential remediation strategies., (Copyright © 2019 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Microbially induced phosphate precipitation (MIPP) is an advanced bioremediation technology to immobilize heavy metals in soil. In this study, an indigenous bacterial strain LRP3, identified as Rahnella sp., was isolated from Cu-contaminated dark brown soil in the mining area. Strain LRP3 could produce phytase and alkaline phosphatase to degrade phytic acid, which released soluble phosphate to the bacterial culture. Due to the metabolism of bacterial growth, the pH value of bacterial culture was increased. The minimum inhibitory concentration of Cu (II) to bacterial growth in solution was up to 130 mg/L. The bacterial culture could rapidly precipitate Cu (II) in solution through MIPP. The analysis results of Scanning Electron Microscope-Energy Dispersive Spectroscopy (SEM-EDS), Fourier Transform-Infrared Spectrometer (FTIR), and X-ray Diffraction (XRD) revealed that the precipitate form by bacterial culture was rod-shaped Cu 3 (OH) 3 PO 4 crystal with a diameter of 10 μm. The bacterial culture decreased the content of DTPA-Cu of 83 mg/kg soil in the soil by 58.2%, 61.5% and 75.8% after 5, 10 and 30 days of incubation, respectively, at the temperature of 25 °C. The results indicate that MIPP-based bioremediation by Rahnella sp. LRP3 is a practical, environmental friendly technology for the cleaning-up of copper-contaminated soil., (Copyright © 2019 Elsevier B.V. All rights reserved.)

This study aimed to reveal the specific mechanism of extracellular polymeric substances (EPS) in the bioleaching of copper-bearing sulfide ore by moderately thermophilic bacterium Acidithiobacillus caldus. The bioleaching performance of blank control (BC), planktonic cell deficient (PD), attached cell deficient (AD), and EPS deficient (ED) systems were compared, to investigate the specific functions of "non-contact" and "contact" (including direct contact and, EPS-mediated contact) mechanisms. The detailed mechanics of bioleaching were studied using μx of cell growth, scanning electron microscopy (SEM), X-ray diffraction (XRD), and Fourier transform infrared spectroscopy (FTIR). The μx of cell growth demonstrated that EPS favors planktonic and attached cell growth. SEM observation revealed that intensive micro-pores on slag benefitted from the "EPS-mediated contact" mechanism. XRD identification indicated that additional chemical derivatives were produced via "EPS-mediated contact" mechanism, because of the active iron/sulfur metabolism. FTIR analysis revealed that the absorption peaks of C-O-S, sulfate, and S = O, which are closely associated with sulfur metabolism, have significant influences of EPS secretion. Taken together, the "EPS-mediated contact" mechanism contributed to almost half of the "contact" mechanism efficiency and a quarter of the total bioleaching efficiency. The proposed specific mechanism of EPS can deepen our understanding of similar bioleaching processes., Competing Interests: The authors have declared that no competing interests exist.

Ramie (Boehmeria nivea L.), the oldest fiber crop in China, can also be grown as fodder crop because of its huge biomass production. Moreover, it has the potential to colonize heavy metal-contaminated soils which showed the possibilities of phytoremediation using B. nivea. Therefore, the present study was conducted to investigate the potential of B. nivea for phytoextraction of copper (Cu)-contaminated soil. Moreover, the impact of different concentrations of Cu on growth and antioxidant enzymatic activity by B. nivea were also studied. For this purpose, a pot experiment was conducted to examine the growth, antioxidative response, and localization (distribution) of Cu in B. nivea plant under different Cu concentrations (0, 50, 100, 200, 300, and 400 mg kg -1 soil). Results revealed that B. nivea tolerated up to 100 mg kg -1 Cu concentration without a significant decrease in biomass, but further increase in Cu concentration from 200 to 400 mg kg -1 exhibited a significant reduction in chlorophyll content, fresh and dry biomass, plant height, and number of leaves. It was further observed that B. nivea accumulated more Cu in roots (26 to 53 mg kg -1 ), followed by the leaves (23 to 28 mg kg -1 ) and stems (14 to 21 mg kg -1 ), while the values for both bioaccumulation factor (BAF) and translocation factor (TF) at all treatments were less than 1. Moreover, activities of antioxidative enzymes (superoxide dismutase and peroxidase) were initially increased with the exposure of 50, 100, and 200 mg kg -1 Cu, but decreased by further increasing the Cu concentration to 300 and 400 mg kg -1 indicating the oxidative stress which is manifested by high malondialdehyde (MDA) and proline contents also. Thus, based on results, it can be concluded that B. nivea accumulated relatively low Cu contents in aboveground parts and could be grown as fodder crop for phytoremediation of Cu-contaminated sites.

Selecting plant species, especially woody species, that can tolerate and accumulate high levels of heavy metals is crucial for the purpose of phytoremediation. In the present study, seven willow species/clones were evaluated for their variations in copper (Cu) tolerance, Cu accumulation, and their relative physiological responses, when exposed to different doses of Cu (control, 15, and 120 μM) in a hydroponic system for 40 days. Upon Cu exposure, all tested willow species/clones (Salix, S.) remained relative normal growth, albeit with some visual evidence of Cu toxicity observed. Seven willow species remained relative high total biomass with tolerance index > 0.6 when being exposed to 120 μM Cu, suggesting their high Cu tolerance. Exposure to 120 μM Cu resulted in notable declines (16.3-76.1%) in photosynthesis in all willow species. Increases in the soluble sugar content and decreases in the soluble protein content in the leaves of five willow species (S. integra "Yizhibi", S. jiangsuensis "J172", S. matsudana 14, S. matsudana 25, S. matsudana 89) were found in the 120 μM Cu treatment. The majority of Cu mainly accumulated in the roots, ranging from 1916 to 26,244 mg kg -1 DW. Principal component analysis and membership function analysis suggested that S. matsudana 89 and S. matsudana 25 showed much higher biomass and accumulation ability than the other species. This suggests that these two willow clones could be used as potential candidates for the phytostabilization of Cu in contaminated soils.

Five sediment cores (S1-S5) were collected from the Shima River to determine the bioavailability of trace metals (Cd, Cr, Cu, Ni, and Zn) using the modified European Community Bureau of Reference (BCR) procedure. The toxic effects of polluted sediment were assessed using the LUMISTox ® bioassay with Vibrio fischeri and chemical models such as the toxicity unit (TU) of each metal, sum of all TUs (∑TU), and toxic risk index (TRI). The results showed that Cd, Ni, and Zn were present mainly in the acid-soluble and residual fractions, and the residual fraction of Cr accounted for the majority of the metal content (44%), while Cu was present mainly in the reducible and residual fractions. Cd had a mean enrichment factor (EF) of 15.1 and was considered to be severely enriched, while there was a minor enrichment of Cr and moderately severe enrichment of Zn, Cu, and Ni. From the LUMISTox ® bioassay, an acute TU (TUa) value exceeding 0.4 was found at the upper and middle reach sites and was considered to represent slightly acute toxicity, whereas little acute toxicity was found at the lower reach site. The acid-soluble fraction of trace metals was the geochemical fraction mainly responsible for the acute toxicity of the sediment, and acid-soluble Zn and Ni were identified as important contributors to sediment toxicity., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

At high levels, copper in grape mash can inhibit yeast activity and cause stuck fermentations. Wine yeast has limited tolerance of copper and can reduce copper levels in wine during fermentation. This study aimed to understand copper tolerance of wine yeast and establish the mechanism by which yeast decreases copper in the must during fermentation. Three strains of Saccharomyces cerevisiae (lab selected strain BH8 and industrial strains AWRI R2 and Freddo) and a simple model fermentation system containing 0 to 1.50 mM Cu2+ were used. ICP-AES determined Cu ion concentration in the must decreasing differently by strains and initial copper levels during fermentation. Fermentation performance was heavily inhibited under copper stress, paralleled a decrease in viable cell numbers. Strain BH8 showed higher copper-tolerance than strain AWRI R2 and higher adsorption than Freddo. Yeast cell surface depression and intracellular structure deformation after copper treatment were observed by scanning electron microscopy and transmission electron microscopy; electronic differential system detected higher surface Cu and no intracellular Cu on 1.50 mM copper treated yeast cells. It is most probably that surface adsorption dominated the biosorption process of Cu2+ for strain BH8, with saturation being accomplished in 24 h. This study demonstrated that Saccharomyces cerevisiae strain BH8 has good tolerance and adsorption of Cu, and reduces Cu2+ concentrations during fermentation in simple model system mainly through surface adsorption. The results indicate that the strain selected from China's stress-tolerant wine grape is copper tolerant and can reduce copper in must when fermenting in a copper rich simple model system, and provided information for studies on mechanisms of heavy metal stress.

Contamination by heavy metals is a major environmental problem worldwide and microbial bioremediation is an efficient method for removing this type of pollution. The plant enzymephytochelatin synthase (PCS, also known as glutathione g-glutamylcysteinyltransferase, EC2.3.2.15) involved in the synthesis of phytochelatins (PCs), which are metal-binding cysteine-rich peptides, has a major role in the detoxification of heavy metals in plants. Expression of the PcPCS1 gene from the bean pear (Pyrus calleryana Dcne.) was induced after cadmium and copper treatments. However, functional analysis of this gene in vivo has not been reported. And it is or not suitable for bioremediation also needs to be assessed. In this study, we found Escherichia coli with over-expressed PcPCS1 had enhanced tolerance to cadmium, copper, sodium, and mercury. E. colicells transformed with pPcPCS1 was found to survive in solid M9 medium containing 2.0 mM Cd(2+), 4.0 mM Cu(2+). 4.5% (w/v) Na+, or 200 μ MHg(2+). Moreover, the growth curve showed 1.5 mM Cd(2+), 2.5 mM Cu(2+), 3.5% (w/v) Naþ, and 100 μ MHg(2+) had no effect on the growth of the E. coli cells transformed with pPcPCS1. Also, we found the contents of PCs and the accumulation of cadmium,copper, sodium, and mercury ions were enhanced in the recombinant E. coli strain Rosetta(TM) (DE3).These results suggested the PcPCS1 gene might be a candidate for heavy metal bioremediation via recombinant bacteria.

Background and Aims: Wilson's disease (WD), characterized by a disorder of copper metabolism, is an inherited autosomal recessive disease caused by mutations in the ATP7B gene., Methods: To explore genotype-phenotype correlations in Chinese WD patients and to evaluate the frequency of the ATP7B mutations, we described 77 clinically and biochemically confirmed WD patients and detected mutations in ten WD families from southwestern China. Clinical features were presented and all the exons of the ATP7B gene were screened., Results: The appearance of Kayser-Fleischer (K-F) rings was closely related to onset age, particularly before 10 years old. For those patients with predominantly neurological symptoms, MRI was the most sensitive and preferred examination. Eight mutations of the ATP7B gene were detected including seven reported mutations (c.2302dup, c.2304delC, c.2333 G>T, c.2621 C>T, c.2755 C>G, c.2975 C>T and c.1366 G>C) and four novel mutations (c.3446 G>A, c.3767insCA, c.3406 G>A and c.3700delG). c.2333 G>T was detected in 6/20 alleles (30%), accounting for the largest proportion, which could be regarded as a mutation hotspot in this region., Conclusions: Our study extends the mutation spectrum of ATP7B and analyzes the relationship between mutations in the ATP7B gene and clinical findings of WD., (Copyright © 2015 IMSS. Published by Elsevier Inc. All rights reserved.)

The article presents an analysis of DNA sequences of ATP7B gene exon 8-exon 9 and exon 10-exon 12 sections in Chinese Han patients with hepatolenticular degeneration or Wilson's disease (WD). The authors report that they performed polymerase chain reaction amplification of said DNA sequences from genomic DNA extracted from 50 WD patients and 20 normal controls, all through direct bidirectional sequencing. One of their findings is that ATP7B gene intron eight mutation is a possible pathogenic mutation associated with WD pathogenesis.

Trace elements play an important role in maintaining the normal metabolic and immune function. The onset of recurrent respiratory tract infection (RRI) is associated with the immune function, genetic factors and nutritional status. However, the association between the levels of trace elements and RRI remains inconclusive. We aimed to investigate the alterations of hair levels of zinc (Zn), copper (Cu) and iron (Fe) in Chinese children with RRI by performing a meta-analysis. A predefined electronic databases search was performed to identify eligible studies for the analysis of hair Zn, Cu or Fe levels in Chinese children with RRI. Thirteen studies were included. RRI patients displayed significantly lower levels of hair Zn (13 studies, random effects SMD: - 1.215, 95% CI: - 1.704 to - 0.725, p < 0.0001), Cu (11 studies, random effects SMD: - 0.384, 95% CI: - 0.717 to - 0.052, p = 0.023) and Fe (12 studies, random effects SMD: - 0.569, 95% CI: - 0.827 to - 0.312, p < 0.0001) compared with controls. No evidence of publication bias was observed. Sensitivity analysis did not change the results significantly. In conclusion, the deficiency of Zn, Cu and Fe may be contributing factors for the susceptibility of RRI in Chinese children. However, more studies in different ethnicities should be performed in the future.

Metal pollution has been widely reported in mangrove wetlands; however, the mechanisms involved in metal detoxification by mangroves are still poorly understood. This study aimed to investigate the possible function of root anatomy and lignification/suberization on metal uptake and tolerance in seedlings of six species of mangroves. The results revealed that the three rhizophoraceous species (Bruguiera gymnorrhiza (L.) Poir, Kandelia obovata Sheue, Liu & Yong and Rhizophora stylosa Griff) consistently exhibited higher metal tolerances than the three pioneer species (Aegiceras corniculatum (Linn.) Blanco, Acanthus ilicifolius L. and Avicennia marina (Forsk.) Viern.). Moreover, metal-tolerant species often exhibited a thick exodermis with high lignification and suberization. The tolerance indices of the mangroves were found to be positively correlated with the amounts of lignin and suberin deposition within the exodermal cell walls. The observed metal uptake by the excised roots further illustrated that a lignified/suberized exodermis directly delayed the entry of metals into the roots, and thereby contributed to a higher tolerance to heavy metals. In summary, the present study proposes a barrier property of the lignified/suberized exodermis in dealing with the stresses of heavy metals, such that the mangroves which possessed more extensive lignification/suberization within the exodermis appeared to exhibit higher metal tolerance., (© The Author 2014. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oup.com.)

Sinorhizobium meliloti CCNWSX0020, isolated from root nodules of Medicago lupulina growing in gold mine tailings in the northwest of China, displayed both copper resistance and growth promotion of leguminous plants in copper-contaminated soil. Nevertheless, the genetic and biochemical mechanisms responsible for copper resistance in S. meliloti CCNWSX0020 remained uncharacterized. To investigate genes involved in copper resistance, an S. meliloti CCNWSX0020 Tn5 insertion library of 14,000 mutants was created. Five copper-sensitive mutants, named SXa-1, SXa-2, SXc-1, SXc-2, and SXn, were isolated, and the disrupted regions involved were identified by inverse PCR and subsequent sequencing. Both SXa-1 and SXa-2 carried a transposon insertion in lpxXL (SM0020&#95;18047), encoding the LpxXL C-28 acyltransferase; SXc-1 and SXc-2 carried a transposon insertion in merR (SM0020&#95;29390), encoding the regulatory activator; SXn contained a transposon insertion in omp (SM0020&#95;18792), encoding a hypothetical outer membrane protein. The results of reverse transcriptase PCR (RT-PCR) combined with transposon gene disruptions revealed that SM0020&#95;05862, encoding an unusual P-type ATPase, was regulated by the MerR protein. Analysis of the genome sequence showed that this P-type ATPase did not contain an N-terminal metal-binding domain or a CPC motif but rather TPCP compared with CopA from Escherichia coli. Pot experiments were carried out to determine whether growth and copper accumulation of the host plant M. lupulina were affected in the presence of the wild type or the different mutants. Soil samples were subjected to three levels of copper contamination, namely, the uncontaminated control and 47.36 and 142.08 mg/kg, and three replicates were conducted for each treatment. The results showed that the wild-type S. meliloti CCNWSX0020 enabled the host plant to grow better and accumulate copper ions. The plant dry weight and copper content of M. lupulina inoculated with the 5 copper-sensitive mutants significantly decreased in the presence of CuSO4.

Three kinds of samples (acid mine drainage, coal mine wastewater, and thermal spring) derived from different sites were collected in China. Thereafter, these samples were combined and then inoculated into a basal salts solution in which different substrates (ferrous sulfate, elemental sulfur, and chalcopyrite) served as energy sources. After that, the mixed cultures growing on different substrates were pooled equally, resulting in a final mixed culture. After being adapted to gradually increasing pulp densities of chalcopyrite concentrate by serial subculturing for more than 2 years, the final culture was able to efficiently leach the chalcopyrite at a pulp density of 20% (wt/vol). At that pulp density, the culture extracted 60.4% of copper from the chalcopyrite in 25 days. The bacterial and archaeal diversities during adaptation were analyzed by denaturing gradient gel electrophoresis and constructing clone libraries of the 16S rRNA gene. The results show that the culture consisted mainly of four species, including Leptospirillum ferriphilum, Acidithiobacillus caldus, Sulfobacillus acidophilus, and Ferroplasma thermophilum, before adapting to a pulp density of 4%. However, L. ferriphilum could not be detected when the pulp density was greater than 4%. Real-time quantitative PCR was employed to monitor the microbial dynamics during bioleaching at a pulp density of 20%. The results show that A. caldus was the predominant species in the initial stage, while S. acidophilus rather than A. caldus became the predominant species in the middle stage. F. thermophilum accounted for the greatest proportion in the final stage.

Using bio-disturbed sulphide to trace the mobility and transformation of Cu, Pb, Ni and Zn in the sediments of the Spartina alterniflora-dominated salt marsh in the Yangtze River Estuary, measurements were made of the seasonal variations of acid-volatile sulphide (AVS) and of the simultaneously extracted metals (SEM) in the rhizosphere sediments. Microcosm incubation experiments recreating flooding conditions were conducted to evaluate the effect of AVS and other metal binding phases upon the dynamics of Cu, Pb, Ni and Zn in the salt marsh sediments. The results demonstrate that the ratio values of SEM/AVS have a significant seasonal variation in the rhizosphere sediments and that the anoxic conditions in the sediments were likely enhanced by S. alterniflora during the summer and autumn compared with the anoxic conditions resulting from the native species Phragmites australis and Scirpus mariqueter. The incubation experiments suggest that Fe(III) and Mn(IV/III) (hydr)oxides provide important binding sites for heavy metals under oxic conditions, and sulphide provides important binding sites for the Cu and Pb under anoxic conditions. Our observations indicate that the mobility of heavy metals in the salt marsh sediments is strongly influenced by biogeochemical redox processes and that the invasive S. alterniflora may increase the seasonal fluctuation in heavy metal bioavailability in the salt marsh ecosystem.

The bioleachings of chalcopyrite ore were compared after inoculating different cultures enriched from the original acid mine drainage sample. The results showed that the higher bioleaching performance was achieved for inoculation with the enrichment D (0.5 % S, 2 % iron and 1 % chalcopyrite) compared to other enrichment systems. The generated ferric precipitation during bioleaching had a key influence on the final copper extraction. After enrichment, higher ratio of iron-oxidizer and higher ratio of sulfur-oxidizer existed in enrichment B and C, respectively. These caused the different bioleaching behaviours from other systems. Maintaining a suitable equilibrium between iron- and sulfur-oxidizers is significant to decrease ferric precipitation or postpone its formation, finally prolong efficient bioleaching period and improve copper extraction.

Pot experiments and laboratory analysis methods were used to investigate the form transformation of additional copper and selenium and their bioavailability for pakchoi (Brassica chinensis) in co-contaminated soil. The results showed that Cu mainly existed in residual bound form, while selenium was present mainly in organic bound and residual form in the uncontaminated soil. In the contaminated soil, copper was mainly bounded to hydrated oxides of iron and manganese, while Se was in exchangeable and carbonate forms. After one month of growing season, Cu tended to transfer into organic bound fractions, while Se tended to bind to hydrated oxides of iron and manganese. The I(R) value of Cu decreased with increasing copper and selenium concentrations, while the I(R) value of Se decreased with increasing Se concentration and had nothing to do with the concentration of exogenous Cu for both before planting and after harvesting of pakchoi. The parameters estimated by S curve fitting indicated that suitable amount of Se (< or = 10 mg x kg(-1)) could promote the Cu uptake by pakchoi, and certain amount of Cu (< or = 400 mg x kg(-1)) could promote Se absorption by pakchoi. Both the I(R), values for Cu and Se had similar trends as the Cu, Se concentrations in pakchoi, which meant that the I(R) value could be used to evaluate the bioavailability of heavy metals in soil. Partial correlation analysis showed that Cu and Se in exchangeable and organic bound forms in soil had better bioavailability for pakchoi. Therefore, the I(R) value of elements in soils and the change of elements before planting and after harvesting of pakchoi can be used as indicators for evaluating the bioavailability of heavy metals.

In this study, dissolved organic matter (DOM) was extracted from earthworm casts and from the cattle manure with which the earthworms were fed, and a water culture experiment was conducted to study the effects of the DOM on the copper (Cu2+) absorption by ryegrass in the presence of different concentration Cu2+ (0, 5 and 10 mg x L(-1)). With the increasing concentration of Cu2+ in the medium, there was a gradual decrease in the dry mass of ryegrass shoots and roots and in the root length, surface area, volume, and tip number. In the presence of medium Cu2+, DOM increased the biomass of shoots and roots and the root length, surface area, volume, and tip number significantly. DOM reduced the Cu2+ concentration in roots, promoted the Cu2+ translocation from roots to shoots, and significantly increased the Cu2+ accumulation in shoots. The DOM from earthworm casts had better effects than that from cattle manure, and high concentration DOM had better effects than low concentration DOM.

Mining effluents are the main source of metals in the surrounding aquatic environment. The mining district of Purple Mountain has a history of copper mining for more than 30 years, but there is limited investigation of metal bioaccumulation in the aquatic creatures from the Tingjiang river catchment affected by the mining activities. In this study, we collected grass carps (Ctenopharyngodon idellus) from four sites, and analyzed the accumulation of chromium (Cr), nickel (Ni), manganese (Mn), copper (Cu), zinc (Zn), arsenic (As), cadmium (Cd), mercury (Hg), and lead (Pb) in ten tissues (scale, skin, muscle, gill, liver, kidney, fish maw, heart, stomach, and intestine) of the fish samples. Among all tissue samples, the highest concentrations (micrograms per gram wet weight) of Ni (0.263), Cu (69.2), Zn (84.0), As (0.259), Cd (0.640), Hg (0.051), and Pb (0.534) were noted in the liver, gill, and kidney tissues, whereas the highest concentrations of Cr (0.356) and Mn (62.7) were detected in the skin and intestine, respectively. These results gave a better understanding of the variability of metals distribution in different fish tissues. In comparison with the sample sites, metals (especially Mn, Cu, Zn, Ni, and Pb) in liver, gill, kidney, stomach, and intestine showed more inter-site differences than other tissues. The inter-site differences also revealed that site 1 and 2 increased fish uptake of Cu, Zn, Ni, and Pb, which may indicate that the copper mine and urban effluents contributed to high levels of these metals in aquatic environments in site 1 and 2. A potential food safety issue may emerge depending on the mining activities in this region because some metals in a few tissue samples exceeded the guideline values for human consumption of fish.

Total concentration is not a reliable indicator of Cu mobility or its bioavailability in soils. The chemical fraction determines the behavior of Cu in soils and hence its toxicity for terrestrial biota. We used the sequential extraction procedure and barley toxicity tests to examine the transfer of Cu in soils around the Dexing Cu mine and to make an ecological risk assessment of Cu in this area. The bioavailable Cu (exchangeable Cu and carbonate-bound Cu) in each soil profile did not change significantly with soil depth, indicating that the Cu itself was vertically mobile and thus potentially a higher risk to the environment. Cu toxicity and bioaccumulation in plants varied with the soil physicochemical characteristics [e.g., pH, clay content, and cation exchange capacity (CEC)] and the level of bioavailable Cu. Multiple regression analysis revealed that bioavailable Cu and CEC could be used to predict Cu toxicity to barley and that other characteristics (such as soil pH, clay content, or total organic carbon) did not predict the risk of toxicity as well as CEC. For the soil to be suitable for agriculture use, treatment of the local soil contamination with guest soil reclamation and phytoremediation with heavy metal-resistant plants would be necessary.

Air and soil pollution from traffic has been considered as a critical issue to crop production and food safety, however, few efforts have been paid on distinguish the source origin of traffic-related contaminants in rice plant along highway. Therefore, we investigated metals (Pb, Cd, Cr, Zn and Cu) concentrations and stable Pb isotope ratios in rice plants exposed and unexposed to highway traffic pollution in Eastern China in 2008. Significant differences in metals concentrations between the exposed and unexposed plants existed in leaf for Pb, Cd and Zn, in stem only for Zn, and in grain for Pb and Cd. About 46% of Pb and 41% of Cd in the grain were attributed to the foliar uptake from atmosphere, and there were no obvious contribution of atmosphere to the accumulations of Cr, Zn and Cu in grain. Except for Zn, all of the heavy metals in stem were attributed to the root uptake from soil, although significant accumulations of Pb and Cd from atmosphere existed in leaf. This indicated that different processes existed in the subsequent translocation of foliar-absorbed heavy metals between rice organs. The distinct separation of stable Pb isotope ratios among rice grain, leaf, stem, soil and vehicle exhaust further provided evidences on the different pathways of heavy metal accumulation in rice plant. These results suggested that further more attentions should be paid to the atmospheric deposition of heavy metals from traffic emission when plan crop layout for food safety along highway.

Biomass of cyanobacterial bloom from Lake Dianchi was used as a biosorbent for copper removal from aqueous solution. The maximum capacity was found at conditions of pH 4, initial concentration of copper was 10 mg/l and initial dose of biomass was 1.0 g/l. HNO(3) demonstrated the highest desorption efficiency compared with HCl, EDTA, and citric acid. Physical adsorption was assumed not to be the dominant mechanism of biosorption as revealed by scanning electron microscopy and surface area measurement of the biomass. Infrared ray spectra analysis of the biomass suggested that ion-exchange is the principal mechanism for biosorption. Considering the advantages-low cost, easy to collect, and huge in quantity-the Microcystis bloom biomass could be used as a sorbent for copper and other heavy metals removal.

Locus afe&#95;0454 from Acidithiobacillus ferrooxidans (At.ferrooxidans) is annotated as related to copper resistance in The Institute for Genomic Research database. In our study, two At.ferrooxidans strains, 26(#) and DC, with different levels of copper ion resistance were isolated from acid mine drainages at two major copper mines in China, and their copper-resistance capacity was determined. The 26(#) strain had a copper-tolerance level of 0.22 mol/L, whereas the DC strain had a lower copper-tolerance level of 0.04 mol/L. The mutant 26(#) was generated from strain 26(#), and its copper-tolerance level was 0.25 mol/L. Using real-time quantitative reverse transcription polymerase chain reaction, differential expression of the afe&#95;0454 gene during copper ion stress of these three strains was investigated. The results showed that the expression of afe&#95;0454 was increased under copper ion stress, indicating that the afe&#95;0454 gene is sensitive to copper levels. Furthermore, the afe&#95;0454 gene expression ratio varied in the different copper-resistant strains. Gene expression was highest in the highest copper-resistant strain. The deduced amino acid sequence of the afe&#95;0454 gene was 56.87% non-polar, indicating the AFE&#95;0454 protein was hydrophobic. Searching with the AFE&#95;0454 protein in The Institute for Genomic Research database showed that the structure of the copper resistance protein D (CopD), which transports copper ions outside of the cell, had the highest sequence identity (46%). Bioinformatics analysis showed that the AFE&#95;0454 protein has eight transmembrane helixes and was predicted to be localized to the plasma membrane. These results strongly suggested that the AFE&#95;0454 protein is likely a transmembrane protein and might be directly involved in copper ion resistance.

Much attention has been given to heavy metal contamination in coastal waters and on seafood safety induced by bioaccumulation of aquatic organisms in China. To obtain the kinetic characteristics of heavy metal bioaccumulation in Chinese native organisms, the accumulation and elimination of three heavy metals (Cu, Pb, and Cd) in domestic oyster Ostrea plicatula were investigated in both laboratory and field conditions. The kinetic parameters of bioaccumulation, including the uptake rate constant (K(1)), elimination rate constant (K(2)), bioconcentration factors (BCFs), and biological half-life (t(1/2)), were estimated through nonlinear curve fitting using a two-compartment model. The results showed that the Pb and Cd metal concentration data perfectly fitted the two-compartment model, and good agreement was obtained between the predicted data and those obtained by the goodness-of-fit test. However, the Cu data did not fit the model. The modeling results indicated that the Pb and Cd concentrations in O. plicatula could reflect the contamination status of seawater and that O. plicatula is a potentially dependable indicator for monitoring heavy metal (Pb and Cd) contamination and seafood safety in the coastal areas of China.

The microbial communities of leachate from a bioleaching heap located in China were analyzed using the 16S rRNA gene clone library and real-time quantitative PCR. Both methods showed that Leptospirillum spp. were the dominant bacteria, and Ferroplasma acidiphilum were the only archaea detected in the leachate. Clone library results indicated that nine operational taxonomic units (OTUs) were obtained, which fell into four divisions, the Nitrospirae (74%), the gamma-Proteobacteria (14%), the Actinobacteria (6%) and the Euryarchaeota (6%). The results obtained by real-time PCR in some ways were the same as clone library analysis. Furthermore, Sulfobacillus spp., detected only by real-time PCR, suggests that real-time PCR was a reliable technology to study the microbial communities in bioleaching environments. It is a useful tool to assist clone library analysis, to further understand microbial consortia and to have comprehensive and exact microbiological information about bioleaching environments. Finally, the interactions among the microorganisms detected in the leachate were summarized according to the characteristics of these species.

This paper studied the transformation of copper fractions in rhizosphere and non-rhizosphere soils of Festuca arundinacea and Trifolium repens in deserted land of copper tailings. The results showed that the proportion of organic-bound or exchangeable contents to the total contents of copper in rhizosphere increased by 9.81 and 10.42%, while the contents of carbonate-bound or Fe-Mn oxides-bound decreased by 2.96 and 1.82%, respectively. The growth and absorption of F. arundinacea and T. repens accelerated the transformation of copper fractions in rhizosphere soils. The effect of root system of T. repens was prominently greater than that of F. arundinacea.

To assess the feasibility of using animal excrement to biomonitor the extent of heavy metal contamination in the marine environment, concentrations of mercury (Hg), lead (Pb), copper (Cu) and zinc (Zn) in the fresh excrement of seabirds and marine mammals, along with other biomaterials, from the Arctic, Antarctica (West and East), and Xisha Archipelago of the South China Sea were determined. Results show that the excrement of marine animals at higher trophic levels generally contained high levels of Hg, demonstrating the biomagnification of Hg through food chains in different remote regions. Significant variations in metal accumulation in the excrements were observed among the distinctive geographical areas, with the highest Hg concentration in Xisha Archipelago and the highest Pb concentration in the Arctic, which reflects different levels of air metal pollution at various sampling locations. Concentrations of Cu in the excrements primarily correlate to the geochemical background levels in the regions. High Cu concentrations were found near the Great Wall Station in West Antarctica where a copper mineralized belt exists. No clear spatial variation pattern was found for Zn accumulation in the excrement. This study shows that animal excrement can be used as bioindicators for the level of metal contamination in the marine environment, with the advantages of easy sampling, accurate detection (i.e., with high levels of metal accumulation), and reconstructing historical metal contamination trends by long-term monitoring of sedimentary excrements.

Two populations of Elsholtzia haichowensis, which are from an uncontaminated site (Hong'an) and a Cu mine site (Tonglushan) in Hubei province, China, were studied for ATPase activities and Cu accumulation of roots under hydroponic conditions. The root tolerance indices were markedly higher in Tonglushan population than in Hong'an population under Cu stress conditions. The root Cu concentration in Tonglushan population was significantly lower than that in Hong'an population in all Cu treatments. The Cu-stimulated ATPase activity increased significantly under 1 microM Cu, reached a peak at 5 microM Cu and then, began to decrease. However, no significant change of ATPase activity was observed in Hong'an population except for an obvious decrease in 80 microM Cu treatment. These results showed that Cu-ATPase might exist in the root plasma membrane of Tonglushan population and play a possible role in resistance to copper., ((Copyright) 2008 Wiley Periodicals, Inc.)

Human saliva offers a unique noninvasive approach for populational study. Purposes of this study were to investigate the feasibility of using saliva manganese (Mn) concentration as a biomarker of Mn exposure among career welders and to study the variations of Mn, copper (Cu), zinc (Zn), cadmium (Cd), and lead (Pb) in saliva as affected by the welding profession. Forty-nine male welders, of whom 28 were in the low exposed group and 21 in the high exposed group, were recruited. Control subjects were 33 military soldiers without metal exposure. Ambient Mn levels in breathing zones were 0.01, 0.24 and 2.21mg/m(3) for control, low, and high exposed groups, respectively. Saliva samples were collected to quantify metals by inductive coupled plasma mass spectrometer (ICP-MS). Saliva concentrations of Mn and Cu were significantly higher in welders than in controls (p<0.01); the variation in saliva levels appeared likely to be associated with airborne Mn levels among study populations. Saliva levels of Zn were significantly lower in welders than in controls (p<0.05), while Cd and Pb levels in saliva were unchanged. Significant associations were observed between saliva and serum for Mn (r=0.575, p<0.05) and Cu (r=0.50, p<0.05). Moreover, saliva Mn concentrations were higher among welders with 5-10years of employment than those with less than 5years of employment. Linear regression analysis revealed a significant correlation between saliva Mn and Cu and between saliva Mn and Zn. Taken together, these data suggest that Mn concentrations in saliva appear reflective of welders' exposure to airborne Mn and their years of welding experience, respectively. Elevated Mn levels among welders may alter the homeostasis of Cu and Zn.

The study on the distribution, accumulation, and seasonal dynamics of Cu and Zn in shrub-marsh plants Salix rosmarinifolia, Salix pentandra, Carex caespitosa and Carex schmidtii in mountainous areas of Northeast China showed that the Cu concentration in test plants varied from 6 to 12 mg x kg(-1), and its distribution was in the sequence of root > stem > leaf in S. rosmarinifolia and S. pentandra, and of stem > leaf > root in C. caespitosa and C. schmidtii, suggesting that Cu was mainly accumulated in the root of shrubs and the stem or leaf of Carex. Shrubs and Carex had less difference in their Cu concentration. The Zn concentration in test plants was 30-250 mg x kg(-1), and its distribution was in the sequence of leaf > stem > root in S. rosmarinifolia and S. pentandra, and of root > stem > leaf in C. caespitosa and C. schmidtii, indicating that Zn was mainly accumulated in the leaf of shrubs and the root of Carex. Shrubs had a higher Zn concentration than Carex. The accumulation coefficient of Zn in the organs of S. rosmarinifolia and S. pentandra was higher than 1.45, suggesting a good Zn-accumulation ability of these plants. The Cu and Zn concentrations in the aboveground parts of the four plants were higher during the initial growth period and then fluctuated to decrease with season, while those in roots were all higher both in the initial and in the late growth periods.

The accumulation of cadmium (Cd) and copper (Cu) in the polychaete Hediste japonica exposed to the mixture of Cd (or Cu) and petroleum hydrocarbons (PHCs) was investigated and compared with that exposed to single Cd (or Cu). The increased bioavailability of Cd or Cu with exposure concentrations resulted in an increase in the accumulation and net accumulation rate of Cd or Cu during single metal exposure. The net accumulation rate of Cd increased, but the net accumulation rate of Cu decreased with exposure time during single metal exposure, suggesting that H. japonica could actively regulate Cu burden in their body by inhibition of absolute uptake or promotion of excretion. The interactions between Cd (or Cu) and PHCs had complicated influences on the net accumulation rate of Cd and Cu in H. japonica under the condition of the binary mixture, which are dependent on their concentration combinations and exposure time.

Daily exposures to chemicals in drinking water are of importance in risk assessment to human health. However, exposure assessment of chemicals in drinking water has seldom been performed in China. Until now, no such a detailed exposure assessment was reported in China. The aim of the present paper is to describe the scenes of daily exposure to Cu and Zn in drinking water by the population of Shanghai, China in the summer of 2003. A total of 163 volunteers and/or homes were selected to be statistically representative of the population of Shanghai, China. The Modified Consumption Habit Exposure Model (CHEM) method combined with the measurement of metals concentrations in stagnant water and actual ingested water was used to describe the daily exposure. The Consumption Habit Exposure Model method results showed that water consumption ranged from 1.7 to 2.2 L/day, varied with different age, gender and work location groups. Chemical results showed that the average Cu and Zn concentrations in stagnant water were much higher than those in actual ingested water (13.5 microg/L and 1.55 mg/L vs. 10.8 microg/L and 0.29 mg/L). Exposure assessment results also showed that volunteers had the highest exposure from the time of after breakfast to before dinner, and had the lowest exposure during the 15 minutes before sleeping. It was also showed that there were few volunteers directly exposed to stagnant water, although six hours stagnant time can be reached in most of the homes in Shanghai in the morning. The mean daily exposures to Cu and Zn were 0.36 microg/kg/day and 0.01 mg/kg/day. Daily intakes of Cu and Zn from drinking water by Shanghai people were 1.8% and 4.4% of the World Health Organization recommendations for minimum total intakes of Cu and Zn for adults, respectively.

Pot and field experiments were conducted to evaluate bioavailability of Cu in contaminated paddy soil (PS) and phytoremediation potential by Elsholtzia splendens as affected by soil amendments. The results from pot experiment showed that organic manure (M) applied to the PS not only remarkably raised the H2O exchangeable Cu, which were mainly due to the increased exchangeable and organic fractions of Cu in the PS by M, but also stimulated plant growth and Cu accumulation in E. splendens. At M application rate of 5.0%, shoot Cu concentration in the plant increased by four times grown on the PS, so as to the elevated shoot Cu accumulation by three times as compared to the control. In the field trial, soil amendments by M and furnace slag (F), and soil preparations like soil capping (S) and soil discing (D) were performed in the PS. Soil capping and discing considerably declined total Cu in the PS. Application of M solely or together with F enhanced plant growth and increased H2O exchangeable Cu levels in the soil. The increased extractability of Cu in the rhizosphere of E. splendens was noted, which may have mainly attributed to the rhizospheric acidification and chelation by dissolved organic matter (DOM), thus resulting in elevating Cu uptake and accumulation by E. splendens. Amendments with organic manure plus furnace slag (MF) to the PS caused the highest exactable Cu with saturated H2O in the rhizospheric soil of E. splendens after they were grown for 170 days in the PS, thus achieving 1.74 kg Cu ha(-1) removal from the contaminated soil by the whole plant of E. splendens at one season, which is higher than those of the other soil treatments. The results indicated that application of organic manure at a proper rate could enhance Cu bioavailability and increase effectiveness of Cu phytoextraction from the contaminated soil by the metal-tolerant and accumulating plant species (E. splendens).

Certain features of Wilson's disease (WD) in Asia have been found to be different from those in other continents. The higher prevalence rate in Japan is presumably due to a higher consanguinity rate. In Chinese there is a tight linkage between WD and two gene loci for esterase D and retinoblastoma in the long arm of chromosome 13. The high proportion of patients with hepatic presentation accounts for early onset of WD in the Japanese and Chinese series. Skeletal involvement, leg hyperpigmentation, dark complexion, amenorrhea, epileptic seizures, and cerebral white matter degeneration are relatively more common among WD patients in Asia. Excessive copper in the liver appears to have a protective effect against hepatocellular carcinoma and type B hepatitis. Electrophysiological studies suggest widespread functional disturbances of the CNS in WD. Side-effects from penicillamine are rather frequent and often lead to interruption of the therapy. Trien is found to be effective without adverse reactions. Oral zinc therapy may be a suitable alternative for long-term management of WD patients in developing Asian countries.