Accumulation and effects of copper on aquatic macrophytes Potamogeton pectinatus L.: Potential application to environmental monitoring and phytoremediation.

Science.gov (United States)

Costa, Marcela Brandão; Tavares, Francesca Valêncio; Martinez, Claudia Bueno; Colares, Ioni Gonçalves; Martins, Camila de Martinez Gaspar

2018-07-15

This study investigated the ability of Potamogeton pectinatus L. to accumulate copper and its effects on plants. In accumulation tests, macrophytes were exposed (96 h) to different copper concentrations (0-1000 µM) and the metal was measured in media and plant tissues (roots, stems and leaves) to determine the bioconcentration factor (BCF). Plants accumulated high concentrations of copper in a dose-dependent manner and roots was the main organ for copper accumulation. However, the more copper increased in water, the more BCF values decreased. It may be due to either saturation of copper uptake or down-regulation of metal uptake by plants. In the physiological and morphological analyses, plants were kept (96 h) in Hoagland nutrient solution without copper, in full Hoagland solution (0.5 µM Cu) and in Hoagland medium with copper from 1 to 100 µM. The absence and the presence of copper above to 1 µM inhibited photosynthesis. Chlorophylls and carotenoid levels also decreased with the excess of copper, a fact that may have affected the photosystem II-dependent of chlorophyll and caused photosynthesis suppression. Only macrophytes at 10 µM Cu showed decrease in length and number of leaves on the 10th day of the test, when they died. Chlorosis and necrosis were observed in control groups and groups with extra copper, but not in Hoalgand group. Overall, the macrophyte P. pectinatus can be considered a suitable plant for monitoring environments contaminated by copper, based on results of copper accumulation in the plant, decrease in pigment concentration and presence of chlorosis and necrosis. However, values of BCF based on fresh water tissues was not proper to indicate the use of P. pectinatus for cleaning environments contaminated by copper. Copyright © 2018 Elsevier Inc. All rights reserved.

Spatial Pattern of Copper Phosphate Precipitation Involves in Copper Accumulation and Resistance of Unsaturated Pseudomonas putida CZ1 Biofilm.

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Chen, Guangcun; Lin, Huirong; Chen, Xincai

2016-12-28

Bacterial biofilms are spatially structured communities that contain bacterial cells with a wide range of physiological states. The spatial distribution and speciation of copper in unsaturated Pseudomonas putida CZ1 biofilms that accumulated 147.0 mg copper per g dry weight were determined by transmission electron microscopy coupled with energy dispersive X-ray analysis, and micro-X-ray fluorescence microscopy coupled with micro-X-ray absorption near edge structure (micro-XANES) analysis. It was found that copper was mainly precipitated in a 75 μm thick layer as copper phosphate in the middle of the biofilm, while there were two living cell layers in the air-biofilm and biofilm-medium interfaces, respectively, distinguished from the copper precipitation layer by two interfaces. The X-ray absorption fine structure analysis of biofilm revealed that species resembling Cu₃(PO₄)₂ predominated in biofilm, followed by Cu-Citrate- and Cu-Glutathione-like species. Further analysis by micro-XANES revealed that 94.4% of copper were Cu₃(PO₄)₂-like species in the layer next to the air interface, whereas the copper species of the layer next to the medium interface were composed by 75.4% Cu₃(PO₄)₂, 10.9% Cu-Citrate-like species, and 11.2% Cu-Glutathione-like species. Thereby, it was suggested that copper was initially acquired by cells in the biofilm-air interface as a citrate complex, and then transported out and bound by out membranes of cells, released from the copper-bound membranes, and finally precipitated with phosphate in the extracellular matrix of the biofilm. These results revealed a clear spatial pattern of copper precipitation in unsaturated biofilm, which was responsible for the high copper tolerance and accumulation of the biofilm.

Accumulation of copper and zinc by balanus amphitrite in a tropical estuary

Digital Repository Service at National Institute of Oceanography (India)

Anil, A.C.; Wagh, A.B.

@iBalanus amphitrite@@ (Cirripedia: Thoracica) a dominant fouling organism was analysed to assess the accumulation potentialities of copper and zinc. It was observed that it could accumulate Cu to 864.77 mu g g-1 dry wt. and Zn to 1937.50 mu g g-1...

Multielemental accumulation and its intracellular distribution in tissues of some aquatic birds

International Nuclear Information System (INIS)

Nam, Dong-Ha; Anan, Yasumi; Ikemoto, Tokutaka; Tanabe, Shinsuke

2005-01-01

This study was aimed at determining multielemental concentration and its intracellular distribution in selected tissues of cormorant and waterfowl species. Non-essential elements such as Hg, Tl, Cd, Pb and V in tissues were generally consistent with those in ingested items, indicating the significance of food sources of non-essential metal accumulation in great cormorants and mallards. Great cormorants and four waterfowl species examined reflected natural background levels of toxic metals such as Cd, Hg and Pb as well as some essential elements, indicating no specific metal exposure from local sources. Most of Cu, Zn, Se, Rb, Ag, Cd, Cs, and Hg contents were present in the hepatocytosolic fraction, whereas a large percentage of V and Mo were present in insoluble fraction in great cormorant, mallard, and spot-billed duck. The major role of these subcelluar fractions in elemental regulation accounts for the high percentage contribution of each cellular fraction to the total metal contents. Cadmium and Cu are chiefly sequestered through binding to metallothioneins (MTs) of hepatocytosolic fraction in these three avian species. Both MTs and high-molecular-weight substance (HMWS) for Zn and low-molecular-weight substance (LMWS) for Rb were also involved in their sequestration in cytosolic fractions. Relatively different species-specific cytosolic substances were responsible for varying degrees of Ag, Mn, and Co accumulation. It is worth noting that these intracellular metal levels in birds are closely regulated by metal-associated cellular constituents. Therefore, risk assessment studies of metal accumulation in such wild birds should take intracellular metal distribution and specific cellular constituents into account

Anaerobic Copper Toxicity and Iron-Sulfur Cluster Biogenesis in Escherichia coli.

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Tan, Guoqiang; Yang, Jing; Li, Tang; Zhao, Jin; Sun, Shujuan; Li, Xiaokang; Lin, Chuxian; Li, Jianghui; Zhou, Huaibin; Lyu, Jianxin; Ding, Huangen

2017-08-15

While copper is an essential trace element in biology, pollution of groundwater from copper has become a threat to all living organisms. Cellular mechanisms underlying copper toxicity, however, are still not fully understood. Previous studies have shown that iron-sulfur proteins are among the primary targets of copper toxicity in Escherichia coli under aerobic conditions. Here, we report that, under anaerobic conditions, iron-sulfur proteins in E. coli cells are even more susceptible to copper in medium. Whereas addition of 0.2 mM copper(II) chloride to LB (Luria-Bertani) medium has very little or no effect on iron-sulfur proteins in wild-type E. coli cells under aerobic conditions, the same copper treatment largely inactivates iron-sulfur proteins by blocking iron-sulfur cluster biogenesis in the cells under anaerobic conditions. Importantly, proteins that do not have iron-sulfur clusters (e.g., fumarase C and cysteine desulfurase) in E. coli cells are not significantly affected by copper treatment under aerobic or anaerobic conditions, indicating that copper may specifically target iron-sulfur proteins in cells. Additional studies revealed that E. coli cells accumulate more intracellular copper under anaerobic conditions than under aerobic conditions and that the elevated copper content binds to the iron-sulfur cluster assembly proteins IscU and IscA, which effectively inhibits iron-sulfur cluster biogenesis. The results suggest that the copper-mediated inhibition of iron-sulfur proteins does not require oxygen and that iron-sulfur cluster biogenesis is the primary target of anaerobic copper toxicity in cells. IMPORTANCE Copper contamination in groundwater has become a threat to all living organisms. However, cellular mechanisms underlying copper toxicity have not been fully understood up to now. The work described here reveals that iron-sulfur proteins in Escherichia coli cells are much more susceptible to copper in medium under anaerobic conditions than they

Effects of Copper Oxide Nanoparticles on Antioxidant Enzyme Activities and on Tissue Accumulation of Oreochromis niloticus.

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Tunçsoy, Mustafa; Duran, Servet; Ay, Özcan; Cicik, Bedii; Erdem, Cahit

2017-09-01

Accumulation of copper oxide nanoparticles (CuO NPs) in gill, liver and muscle tissues of Oreochromis niloticus and its effects on superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPx) activities in gill and liver tissues were studied after exposing the fish to 20 µg/L Cu over 15 days. Copper levels and enzyme activities in tissues were determined using spectrophotometric (ICP-AES and UV) techniques respectively. No mortality was observed during the experiments. Copper levels increased in gill and liver tissues of O. niloticus compared to control when exposed to CuO NPs whereas exposure to metal had no effect on muscle level at the end of the exposure period. Highest accumulation of copper was observed in liver while no accumulation was detected in muscle tissue. SOD, CAT activities decreased and GPx activity increased in gill and liver tissues when exposed to CuO NPs.

Pathogenic adaptations to host-derived antibacterial copper

Science.gov (United States)

Chaturvedi, Kaveri S.; Henderson, Jeffrey P.

2014-01-01

Recent findings suggest that both host and pathogen manipulate copper content in infected host niches during infections. In this review, we summarize recent developments that implicate copper resistance as an important determinant of bacterial fitness at the host-pathogen interface. An essential mammalian nutrient, copper cycles between copper (I) (Cu+) in its reduced form and copper (II) (Cu2+) in its oxidized form under physiologic conditions. Cu+ is significantly more bactericidal than Cu2+ due to its ability to freely penetrate bacterial membranes and inactivate intracellular iron-sulfur clusters. Copper ions can also catalyze reactive oxygen species (ROS) generation, which may further contribute to their toxicity. Transporters, chaperones, redox proteins, receptors and transcription factors and even siderophores affect copper accumulation and distribution in both pathogenic microbes and their human hosts. This review will briefly cover evidence for copper as a mammalian antibacterial effector, the possible reasons for this toxicity, and pathogenic resistance mechanisms directed against it. PMID:24551598

Acute toxicity and bio-accumulation of mercury and copper in ...

African Journals Online (AJOL)

The acute toxicity of Mercury and Copper on C. africanus and T. fuscatus and the bio-accumulation potentials of the metals were investigated in laboratory experiments employing standard bio–assay techniques. On the basis of LC50 values, both metals had similar magnitudes of toxicity against C. africanus. However ...

ANTIMICROBIAL ACTIVITY OF COPPER AND ZINC ACCUMULATED BY EASTERN OYSTER AMEBOCYTES

Science.gov (United States)

Fisher, William S. Submitted. Antimicrobial Activity of Copper and Zinc Accumulated by Eastern Oyster Amebocytes. J. Shellfish Res. 54 p. (ERL,GB 1196). The distribution of eastern oysters Crassostrea virginica near terrestrial watersheds has led to a general impression t...

Prediction of the long term accumulation and leaching of copper in Dutch agricultural soils: a risk assessment study

NARCIS (Netherlands)

Groenenberg, J.E.; Römkens, P.F.A.M.; Vries, de W.

2006-01-01

This report describes a model study to assess whether current copper inputs on agricultural land lead to accumulation of copper and in time to an exceedance of a Predicted No Effect Concentration. A copper mass balance model was applied to the whole Netherlands. Future copper concentrations after

COPPER ACCUMULATION IN SOILS AND VEGETATION OF POLLUTED AREA COPŞA MICĂ

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Nicoleta Vrînceanu

2010-01-01

Full Text Available The study carried out in order to estimate the distribution and accumulation of copper in soils and vegetation fromCopşa Mică area used a radial network centered in the source of pollution – S.C. SOMETRA S.A. Copşa Mică. Soil andplant samples taken from the radial nodes of the network were analyzed to determine the content of copper. Values ofcopper content in plant ranged between 4.2 mg/kg and 97 mg/kg. Based on these results has been obtained a regressionequation that estimates the copper content in plants as function of the total copper content in soil. The spontaneousvegetation developed in the investigated area includes plants belonging to the following species: Amaranthusretroflexus, Artemisia vulgaris, Asclepias syriaca, Calamagrostis epigeios, Calamagrostis pseudophragmites, Cynodondactylon, Daucus carota, Equisetum arvense, Phragmites australis, Picris hieracioides, Setaria glauca, Sinapisarvensis, Verbascum phlomoides and Xanthium strumarium. The copper pollution doesn’t represent a major problem inCopşa Mică area.

Proteomic and physiological responses of Kineococcus radiotolerans to copper.

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Christopher E Bagwell

Full Text Available Copper is a highly reactive, toxic metal; consequently, transport of this metal within the cell is tightly regulated. Intriguingly, the actinobacterium Kineococcus radiotolerans has been shown to not only accumulate soluble copper to high levels within the cytoplasm, but the phenotype also correlated with enhanced cell growth during chronic exposure to ionizing radiation. This study offers a first glimpse into the physiological and proteomic responses of K. radiotolerans to copper at increasing concentration and distinct growth phases. Aerobic growth rates and biomass yields were similar over a range of Cu(II concentrations (0-1.5 mM in complex medium. Copper uptake coincided with active cell growth and intracellular accumulation was positively correlated with Cu(II concentration in the growth medium (R(2=0.7. Approximately 40% of protein coding ORFs on the K. radiotolerans genome were differentially expressed in response to the copper treatments imposed. Copper accumulation coincided with increased abundance of proteins involved in oxidative stress and defense, DNA stabilization and repair, and protein turnover. Interestingly, the specific activity of superoxide dismutase was repressed by low to moderate concentrations of copper during exponential growth, and activity was unresponsive to perturbation with paraquot. The biochemical response pathways invoked by sub-lethal copper concentrations are exceptionally complex; though integral cellular functions are preserved, in part, through the coordination of defense enzymes, chaperones, antioxidants and protective osmolytes that likely help maintain cellular redox. This study extends our understanding of the ecology and physiology of this unique actinobacterium that could potentially inspire new biotechnologies in metal recovery and sequestration, and environmental restoration.

Proteomic and physiological responses of Kineococcus radiotolerans to copper.

Science.gov (United States)

Bagwell, Christopher E; Hixson, Kim K; Milliken, Charles E; Lopez-Ferrer, Daniel; Weitz, Karl K

2010-08-26

Copper is a highly reactive, toxic metal; consequently, transport of this metal within the cell is tightly regulated. Intriguingly, the actinobacterium Kineococcus radiotolerans has been shown to not only accumulate soluble copper to high levels within the cytoplasm, but the phenotype also correlated with enhanced cell growth during chronic exposure to ionizing radiation. This study offers a first glimpse into the physiological and proteomic responses of K. radiotolerans to copper at increasing concentration and distinct growth phases. Aerobic growth rates and biomass yields were similar over a range of Cu(II) concentrations (0-1.5 mM) in complex medium. Copper uptake coincided with active cell growth and intracellular accumulation was positively correlated with Cu(II) concentration in the growth medium (R(2)=0.7). Approximately 40% of protein coding ORFs on the K. radiotolerans genome were differentially expressed in response to the copper treatments imposed. Copper accumulation coincided with increased abundance of proteins involved in oxidative stress and defense, DNA stabilization and repair, and protein turnover. Interestingly, the specific activity of superoxide dismutase was repressed by low to moderate concentrations of copper during exponential growth, and activity was unresponsive to perturbation with paraquot. The biochemical response pathways invoked by sub-lethal copper concentrations are exceptionally complex; though integral cellular functions are preserved, in part, through the coordination of defense enzymes, chaperones, antioxidants and protective osmolytes that likely help maintain cellular redox. This study extends our understanding of the ecology and physiology of this unique actinobacterium that could potentially inspire new biotechnologies in metal recovery and sequestration, and environmental restoration.

Molecular mechanism of intracellular lipid accumulation: Suppressive effect of PycnogenolR in liver cells

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Shoichiro Ikuyama

2013-09-01

Full Text Available ABSTRACTCells are physiologically ready to accumulate lipids such as triacylglycerides in the cytoplasm.Five classes of perilipin (PLIN family proteins are known to be involved in the process of intracellular lipid accumulation. PLIN2 is expressed ubiquitously including adipocytes, hepatocytes and macrophages. Over-expression of PLIN2 is demonstrated in the lesions of fatty liver diseases and atherosclerosis. Suppression of PLIN2 expression prevents from developing these pathological conditions in animal models, suggesting that PLIN2 could be a therapeutic target molecule for excessive intracellular lipid accumulation which leads to various metabolic derangements. The PLIN2 gene promoter has two important cis-acting elements in close proximity:AP-1 element which mediates inflammatory signals and PPRE which mediates free fatty acid effect. In NMuLi mouse liver cells, FFA such as oleic acid requires both functional AP-1 and PPRE simultaneously to stimulate the promoter activity, indicating the presence of intimate interaction of inflammatory and metabolic signals on this gene. PycnogenolR, French maritime pine bark extracts, suppressed the oleic acid-induced PLIN2 expression and lipid accumulation in NMuLi cells. We found that PycnogenolR did not suppress the PLIN2 promoter activity or AP-1 binding to DNA. Instead, PycnogenolRfacilitates the PLIN2 mRNA degradation, leading to suppression of lipid accumulation. This effect seems to be independent of antioxidant effect of PycnogenolR.We raise the idea that PLIN2 is a putative target molecule for prevention of pathological condition induced by excessive lipid accumulation, and this class of natural compounds could be putative therapeutic modalities.Key words: PycnogenolR, lipid droplet, perilipin, fatty liver disease

Impact of intracellular chloride concentration on cisplatin accumulation in sensitive and resistant GLC4 cells

NARCIS (Netherlands)

Salerno, Milena; Yahia, Dalila; Dzamitika, Simplice; de Vries, Elisabeth G. E.; Pereira-Maia, Elene; Garnier-Suillerot, Arlette

Resistance to cisplatin [cis-diamminedichloroplatinum(II), CDDP] chemotherapy is a major problem in the clinic. Understanding the molecular basis of the intracellular accumulation of CDDP and other platinum-based anticancer drugs is of importance in delineating the mechanism of resistance to these

Intergenotypic replacement of lyssavirus matrix proteins demonstrates the role of lyssavirus M proteins in intracellular virus accumulation.

Science.gov (United States)

Finke, Stefan; Granzow, Harald; Hurst, Jose; Pollin, Reiko; Mettenleiter, Thomas C

2010-02-01

Lyssavirus assembly depends on the matrix protein (M). We compared lyssavirus M proteins from different genotypes for their ability to support assembly and egress of genotype 1 rabies virus (RABV). Transcomplementation of M-deficient RABV with M from European bat lyssavirus (EBLV) types 1 and 2 reduced the release of infectious virus. Stable introduction of the heterogenotypic M proteins into RABV led to chimeric viruses with reduced virus release and intracellular accumulation of virus genomes. Although the chimeras indicated genotype-specific evolution of M, rapid selection of a compensatory mutant suggested conserved mechanisms of lyssavirus assembly and the requirement for only few adaptive mutations to fit the heterogenotypic M to a RABV backbone. Whereas the compensatory mutant replicated to similar infectious titers as RABV M-expressing virus, ultrastructural analysis revealed that both nonadapted EBLV M chimeras and the compensatory mutant differed from RABV M expressing viruses in the lack of intracellular viruslike structures that are enveloped and accumulate in cisterna of the degranulated and dilated rough endoplasmic reticulum compartment. Moreover, all viruses were able to bud at the plasma membrane. Since the lack of the intracellular viruslike structures correlated with the type of M protein but not with the efficiency of virus release, we hypothesize that the M proteins of EBLV-1 and RABV differ in their target membranes for virus assembly. Although the biological function of intracellular assembly and accumulation of viruslike structures in the endoplasmic reticulum remain unclear, the observed differences could contribute to diverse host tropism or pathogenicity.

Toxicity and deficiency of copper in Elsholtzia splendens affect photosynthesis biophysics, pigments and metal accumulation.

Science.gov (United States)

Peng, Hongyun; Kroneck, Peter M H; Küpper, Hendrik

2013-06-18

Elsholtzia splendens is a copper-tolerant plant species growing on copper deposits in China. Spatially and spectrally resolved kinetics of in vivo absorbance and chlorophyll fluorescence in mesophyll of E. splendens were used to investigate the copper-induced stress from deficiency and toxicity as well as the acclimation to excess copper stress. The plants were cultivated in nutrient solutions containing either Fe(III)-EDTA or Fe(III)-EDDHA. Copper toxicity affected light-acclimated electron flow much stronger than nonphotochemical quenching (NPQ) or dark-acclimated photochemical efficiency of PSIIRC (Fv/Fm). It also changed spectrally resolved Chl fluorescence kinetics, in particular by strengthening the short-wavelength (<700 nm) part of NPQ altering light harvesting complex II (LHCII) aggregation. Copper toxicity reduced iron accumulation, decreased Chls and carotenoids in leaves. During acclimation to copper toxicity, leaf copper decreased but leaf iron increased, with photosynthetic activity and pigments recovering to normal levels. Copper tolerance in E. splendens was inducible; acclimation seems be related to homeostasis of copper and iron in E. splendens. Copper deficiency appeared at 10 mg copper per kg leaf DW, leading to reduced growth and decreased photosynthetic parameters (F0, Fv/Fm, ΦPSII). The importance of these results for evaluating responses of phytoremediation plants to stress in their environment is discussed.

Involvement of indole-3-acetic acid produced by Azospirillum brasilense in accumulating intracellular ammonium in Chlorella vulgaris.

Science.gov (United States)

Meza, Beatriz; de-Bashan, Luz E; Bashan, Yoav

2015-01-01

Accumulation of intracellular ammonium and activities of the enzymes glutamine synthetase (GS) and glutamate dehydrogenase (GDH) were measured when the microalgae Chlorella vulgaris was immobilized in alginate with either of two wild type strains of Azospirillum brasilense or their corresponding indole-3-acetic acid (IAA)-attenuated mutants. After 48 h of immobilization, both wild types induced higher levels of intracellular ammonium in the microalgae than their respective mutants; the more IAA produced, the higher the intracellular ammonium accumulated. Accumulation of intracellular ammonium in the cells of C. vulgaris followed application of four levels of exogenous IAA reported for A. brasilense and its IAA-attenuated mutants, which had a similar pattern for the first 24 h. This effect was transient and disappeared after 48 h of incubation. Immobilization of C. vulgaris with any bacteria strain induced higher GS activity. The bacterial strains also had GS activity, comparable to the activity detected in C. vulgaris, but weaker than when immobilized with the bacteria. When net activity was calculated, the wild type always induced higher GS activity than IAA-attenuated mutants. GDH activity in most microalgae/bacteria interactions resembled GS activity. When complementing IAA-attenuated mutants with exogenous IAA, GS activity in co-immobilized cultures matched those of the wild type A. brasilense immobilized with the microalga. Similarity occurred when the net GS activity was measured, and was higher with greater quantities of exogenous IAA. It is proposed that IAA produced by A. brasilense is involved in ammonium uptake and later assimilation by C. vulgaris. Copyright © 2014 Institut Pasteur. Published by Elsevier Masson SAS. All rights reserved.

Metabolism of manganese, iron, copper, and selenium in calves

International Nuclear Information System (INIS)

Ho, S.Y.

1981-01-01

Sixteen male Holstein calves were used to study manganese and iron metabolism. The calves were fed one of the following diets for 18 days: control, control + iron, control + manganese, and control + iron and manganese. All calves were dosed orally with manganese-54. Tissue concentrations of manganese, iron and manganese-54 were determined. Small intestinal iron was lower in calves fed the high manganese diet than in controls. Tissue manganese-54 was lower in calves fed a high manganese diet. Fecal manganese content increased in calves fed both high manganese and high manganese-high iron diets. Serum total iron was not affected by the dietary treatments. To study the effects of high dietary levels of copper and selenium on the intracellular distributions of these two elements in liver and kidney cytosol, calves were fed one of four diets for 15 days. These were 0 and 100 ppM supplemental copper and 0 and 1 ppM added selenium. The control diet containing 0.1 ppM of selenium and 15 ppM of copper. All calves were orally dosed 48 hrs prior to sacrifice with selenium-75. A high copper diet increased copper concentrations in all intracellular liver fractions and most kidney fractions. Only the effects in the liver were significant. Less copper was found in the mitochondria fractions in liver and kidney of calves fed a high selenium diet. Three major copper-binding protein peaks were separated from the soluble fractions of calf liver and kidney. Peak 1 appeared to be the major copper-binding protein in liver and kidney cytosol of copper-loaded animals. Added selenium alone or in combination with copper accentuated the copper accumulation in this peak. Most of selenium-75 was recovered in the same peak as the copper. The results of this experiment indicated that the large molecular proteins in liver and kidney cytosol of calves play an important role in copper and selenium-75 metabolism

Soluble Moringa oleifera leaf extract reduces intracellular cadmium accumulation and oxidative stress in Saccharomyces cerevisiae.

Science.gov (United States)

Kerdsomboon, Kittikhun; Tatip, Supinda; Kosasih, Sattawat; Auesukaree, Choowong

2016-05-01

Moringa oleifera leaves are a well-known source of antioxidants and traditionally used for medicinal applications. In the present study, the protective action of soluble M. oleifera leaf extract (MOLE) against cadmium toxicity was investigated in the model eukaryote Saccharomyces cerevisiae. The results showed that this extract exhibited a protective effect against oxidative stress induced by cadmium and H2O2 through the reduction of intracellular reactive oxygen species. Interestingly, not only the co-exposure of soluble MOLE with cadmium but also pretreatment of this extract prior to cadmium exposure significantly reduced the cadmium uptake through an inhibition of Fet4p, a low-affinity iron(II) transporter. In addition, the supplementation of soluble MOLE significantly reduced intracellular iron accumulation in a Fet4p-independent manner. Our findings suggest the potential use of soluble extract from M. oleifera leaves as a dietary supplement for protection against cadmium accumulation and oxidative stress. Copyright © 2015 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.

Cathepsin-Mediated Cleavage of Peptides from Peptide Amphiphiles Leads to Enhanced Intracellular Peptide Accumulation

Energy Technology Data Exchange (ETDEWEB)

Acar, Handan [Institute; Department; Samaeekia, Ravand [Institute; Department; Schnorenberg, Mathew R. [Institute; Department; Medical; Sasmal, Dibyendu K. [Institute; Huang, Jun [Institute; Tirrell, Matthew V. [Institute; Institute; LaBelle, James L. [Department

2017-08-24

Peptides synthesized in the likeness of their native interaction domain(s) are natural choices to target protein protein interactions (PPIs) due to their fidelity of orthostatic contact points between binding partners. Despite therapeutic promise, intracellular delivery of biofunctional peptides at concentrations necessary for efficacy remains a formidable challenge. Peptide amphiphiles (PAs) provide a facile method of intracellular delivery and stabilization of bioactive peptides. PAs consisting of biofunctional peptide headgroups linked to hydrophobic alkyl lipid-like tails prevent peptide hydrolysis and proteolysis in circulation, and PA monomers are internalized via endocytosis. However, endocytotic sequestration and steric hindrance from the lipid tail are two major mechanisms that limit PA efficacy to target intracellular PPIs. To address these problems, we have constructed a PA platform consisting of cathepsin-B cleavable PAs in which a selective p53-based inhibitory peptide is cleaved from its lipid tail within endosomes, allowing for intracellular peptide accumulation and extracellular recycling of the lipid moiety. We monitor for cleavage and follow individual PA components in real time using a resonance energy transfer (FRET)-based tracking system. Using this platform, components in real time using a Forster we provide a better understanding and quantification of cellular internalization, trafficking, and endosomal cleavage of PAs and of the ultimate fates of each component.

Copper as a target for prostate cancer therapeutics: copper-ionophore pharmacology and altering systemic copper distribution

Science.gov (United States)

Denoyer, Delphine; Pearson, Helen B.; Clatworthy, Sharnel A.S.; Smith, Zoe M.; Francis, Paul S.; Llanos, Roxana M.; Volitakis, Irene; Phillips, Wayne A.; Meggyesy, Peter M.; Masaldan, Shashank; Cater, Michael A.

2016-01-01

Copper-ionophores that elevate intracellular bioavailable copper display significant therapeutic utility against prostate cancer cells in vitro and in TRAMP (Transgenic Adenocarcinoma of Mouse Prostate) mice. However, the pharmacological basis for their anticancer activity remains unclear, despite impending clinical trails. Herein we show that intracellular copper levels in prostate cancer, evaluated in vitro and across disease progression in TRAMP mice, were not correlative with copper-ionophore activity and mirrored the normal levels observed in patient prostatectomy tissues (Gleason Score 7 & 9). TRAMP adenocarcinoma cells harbored markedly elevated oxidative stress and diminished glutathione (GSH)-mediated antioxidant capacity, which together conferred selective sensitivity to prooxidant ionophoric copper. Copper-ionophore treatments [CuII(gtsm), disulfiram & clioquinol] generated toxic levels of reactive oxygen species (ROS) in TRAMP adenocarcinoma cells, but not in normal mouse prostate epithelial cells (PrECs). Our results provide a basis for the pharmacological activity of copper-ionophores and suggest they are amendable for treatment of patients with prostate cancer. Additionally, recent in vitro and mouse xenograft studies have suggested an increased copper requirement by prostate cancer cells. We demonstrated that prostate adenocarcinoma development in TRAMP mice requires a functional supply of copper and is significantly impeded by altered systemic copper distribution. The presence of a mutant copper-transporting Atp7b protein (tx mutation: A4066G/Met1356Val) in TRAMP mice changed copper-integration into serum and caused a remarkable reduction in prostate cancer burden (64% reduction) and disease severity (grade), abrogating adenocarcinoma development. Implications for current clinical trials are discussed. PMID:27175597

Characterization of copper transport in gill cells of a mangrove crab Ucides cordatus

Energy Technology Data Exchange (ETDEWEB)

Sá, M.G. [Biosciences Institute, Department of Physiology, University of São Paulo, Rua do Matão, Travessa 14, 101, São Paulo 05508-900, SP (Brazil); Zanotto, F.P., E-mail: fzanotto@usp.br [Biosciences Institute, Department of Physiology, University of São Paulo, Rua do Matão, Travessa 14, 101, São Paulo 05508-900, SP (Brazil); Department of Biophysics, Escola Paulista de Medicina, Universidade Federal de Sao Paulo, Rua Três de Maio 100, Sao Paulo 04044-020 (Brazil)

2013-11-15

Highlights: •Copper transport in gill cells of a mangrove crab Ucides cordatus is dependent of calcium. •Copper transport mechanism is ATP-dependent. •Transport was monitored second by second during 300 s. -- Abstract: The branchial epithelium of crustaceans is exposed to the environment and is the first site affected by metal pollution. The aim of this work was to characterize copper (Cu) transport using a fluorescent dye, Phen Green, in gill cells of a hypo-hyper-regulator mangrove crab Ucides cordatus. The results showed that added extracellular CuCl{sub 2} (0, 0.025, 0.150, 0.275, 0.550 and 1.110 μM) showed typical Michaelis–Menten transport for Cu in anterior and posterior gill cells (V{sub max} for anterior and posterior gills: 0.41 ± 0.12 and 1.76 ± 0.27 intracellular Cu in μM × 22.10{sup 4} cells{sup −1} × 300 s{sup −1} respectively and K{sub m} values: 0.44 ± 0.04 and 0.32 ± 0.13 μM, respectively). Intracellular Cu was significantly higher for posterior gill cells compared to anterior gill cells, suggesting differential accumulation for each gill type. Extracellular Ca at 20 mM decreased cellular Cu transport for both anterior and posterior gill cells. Nifedipine and verapamil, calcium channel inhibitors from plasma membrane, decreased Cu transport and affected K{sub m} for both gills. These results could be due to a competition between Cu and Ca. Amiloride, a Na/Ca exchanger inhibitor, as well as bafilomycin, a proton pump inhibitor, caused a decrease of intracellular Cu compared to control. Ouabain and KB-R 7943, acting on Na homeostasis, similarly decreased intracellular Cu in both gill cells. Besides that, gill cells exposed to ATP and Cu simultaneously, showed an increase in intracellular copper, which was inhibited by vanadate, an inhibitor of P-type ATPase. These results suggest either the presence of a Cu-ATPase in crab gill cells, responsible for Cu influx, or the effect of a change in electrochemical membrane potential that

Characterization of copper transport in gill cells of a mangrove crab Ucides cordatus

International Nuclear Information System (INIS)

Sá, M.G.; Zanotto, F.P.

2013-01-01

Highlights: •Copper transport in gill cells of a mangrove crab Ucides cordatus is dependent of calcium. •Copper transport mechanism is ATP-dependent. •Transport was monitored second by second during 300 s. -- Abstract: The branchial epithelium of crustaceans is exposed to the environment and is the first site affected by metal pollution. The aim of this work was to characterize copper (Cu) transport using a fluorescent dye, Phen Green, in gill cells of a hypo-hyper-regulator mangrove crab Ucides cordatus. The results showed that added extracellular CuCl 2 (0, 0.025, 0.150, 0.275, 0.550 and 1.110 μM) showed typical Michaelis–Menten transport for Cu in anterior and posterior gill cells (V max for anterior and posterior gills: 0.41 ± 0.12 and 1.76 ± 0.27 intracellular Cu in μM × 22.10 4 cells −1 × 300 s −1 respectively and K m values: 0.44 ± 0.04 and 0.32 ± 0.13 μM, respectively). Intracellular Cu was significantly higher for posterior gill cells compared to anterior gill cells, suggesting differential accumulation for each gill type. Extracellular Ca at 20 mM decreased cellular Cu transport for both anterior and posterior gill cells. Nifedipine and verapamil, calcium channel inhibitors from plasma membrane, decreased Cu transport and affected K m for both gills. These results could be due to a competition between Cu and Ca. Amiloride, a Na/Ca exchanger inhibitor, as well as bafilomycin, a proton pump inhibitor, caused a decrease of intracellular Cu compared to control. Ouabain and KB-R 7943, acting on Na homeostasis, similarly decreased intracellular Cu in both gill cells. Besides that, gill cells exposed to ATP and Cu simultaneously, showed an increase in intracellular copper, which was inhibited by vanadate, an inhibitor of P-type ATPase. These results suggest either the presence of a Cu-ATPase in crab gill cells, responsible for Cu influx, or the effect of a change in electrochemical membrane potential that could also drive Cu to the gill cell

Hardness and microstructure homogeneity of pure copper processed by accumulative back extrusion

International Nuclear Information System (INIS)

Bazaz, B.; Zarei-Hanzaki, A.; Fatemi-Varzaneh, S.M.

2013-01-01

The present work deals with the microstructure evolution of a pure copper processed by a new severe plastic deformation method. A set of pure copper (99.99%) work-pieces with coarse-grained microstructures was processed by accumulative back extrusion (ABE) method at room temperature. The optical and scanning electron microscopy (SEM) and hardness measurements were utilized to study the microstructural evolution and hardness homogeneity. The results indicated that ABE is a capable process to provide a homogenous grain refined microstructure in pure copper. The observed grain refinement was discussed relying on the occurrence of dynamic restoration processes. The analysis of microstructure and hardness showed outstanding homogeneity improvement throughout the work-pieces as the consecutive ABE passes were applied. The homogeneity improvement was attributed to the propagation of the shear bands and also the heavily deformed regions. A reversing route was also applied in the ABE processing to investigate its effect on the development of microstructural homogeneity. Comparing to the conventional route, the application of the reversing route was found to yield better homogeneity after less passes of the process.

Ultrafine-Grained Precipitation Hardened Copper Alloys by Swaging or Accumulative Roll Bonding

Directory of Open Access Journals (Sweden)

Igor Altenberger

2015-05-01

Full Text Available There is an increasing demand in the industry for conductive high strength copper alloys. Traditionally, alloy systems capable of precipitation hardening have been the first choice for electromechanical connector materials. Recently, ultrafine-grained materials have gained enormous attention in the materials science community as well as in first industrial applications (see, for instance, proceedings of NANO SPD conferences. In this study the potential of precipitation hardened ultra-fine grained copper alloys is outlined and discussed. For this purpose, swaging or accumulative roll-bonding is applied to typical precipitation hardened high-strength copper alloys such as Corson alloys. A detailed description of the microstructure is given by means of EBSD, Electron Channeling Imaging (ECCI methods and consequences for mechanical properties (tensile strength as well as fatigue and electrical conductivity are discussed. Finally the role of precipitates for thermal stability is investigated and promising concepts (e.g. tailoring of stacking fault energy for grain size reduction and alloy systems for the future are proposed and discussed. The relation between electrical conductivity and strength is reported.

Copper-Based Aquatic Algaecide Adsorption and Accumulation Kinetics: Influence of Exposure Concentration and Duration for Controlling the Cyanobacterium Lyngbya wollei.

Science.gov (United States)

Bishop, West M; Lynch, Clayton L; Willis, Ben E; Cope, W Gregory

2017-09-01

Filamentous mat-forming cyanobacteria are increasingly impairing uses of freshwater resources. To effectively manage, a better understanding of control measures is needed. Copper (Cu)-based algaecide formulations are often applied to reactively control nuisance cyanobacterial blooms. This laboratory research assessed typical field exposure scenarios for the ability of Cu to partition to, and accumulate in Lyngbya wollei. Exposure factors (Cu concentration × duration) of 4, 8, 16, 24, 32 h were tested across three aqueous Cu concentrations (1, 2, 4 ppm). Results indicated that internally accumulated copper correlated with control of L. wollei, independent of adsorbed copper. L. wollei control was determined by filament viability and chlorophyll a concentrations. Similar exposure factors elicited similar internalized copper levels and consequent responses of L. wollei. Ultimately, a "concentration-exposure-time" (CET) model was created to assist water resource managers in selecting an appropriate treatment regime for a specific in-water infestation. By assessing the exposure concentration and duration required to achieve the internal threshold of copper (i.e., critical burden) that elicits control, water management objectives can be achieved while simultaneously decreasing the environmental loading of copper and potential for non-target species risks.

Carbon black nanoparticles promote endothelial activation and lipid accumulation in macrophages independently of intracellular ROS production

DEFF Research Database (Denmark)

Cao, Yi; Roursgaard, Martin; Danielsen, Pernille Høgh

2014-01-01

, the concentrations of CB to induce lipid accumulation were lower than the concentrations to promote intracellular ROS production in THP-1a cells. In conclusion, exposure to nano-sized CB induced endothelial dysfunction and foam cell formation, which was not dependent on intracellular ROS production....... and WST-1 assays, especially in THP-1 and THP-1a cells. The CB exposure decreased the glutathione (GSH) content in THP-1 and THP-1a cells, whereas GSH was increased in HUVECs. The reactive oxygen species (ROS) production was increased in all cell types after CB exposure. A reduction of the intracellular...... GSH concentration by buthionine sulfoximine (BSO) pre-treatment further increased the CB-induced ROS production in THP-1 cells and HUVECs. The expression of adhesion molecules ICAM-1 and VCAM-1, but not adhesion of THP-1 to HUVECs or culture dishes, was elevated by CB exposure, whereas these effects...

Intracellular trafficking of a pH-responsive drug metal complex.

Science.gov (United States)

Kheirolomoom, Azadeh; Ingham, Elizabeth S; Commisso, Joel; Abushaban, Neveen; Ferrara, Katherine W

2016-12-10

We previously developed a pH-responsive copper-doxorubicin (CuDox) cargo in lysolipid-based temperature-sensitive liposomes (LTSLs). The CuDox complex is released from the particle by elevated temperature; however, full release of doxorubicin from CuDox requires a reduced pH, such as that expected in lysosomes. The primary goal of this study is to evaluate the cellular uptake and intracellular trafficking of the drug-metal complex in comparison with intact liposomes and free drug. We found that the CuDox complex was efficiently internalized by mammary carcinoma cells after release from LTSLs. Intracellular doxorubicin and copper were 6-fold and 5-fold greater, respectively, after a 0.5h incubation with the released CuDox complex, as compared to incubation with intact liposomes containing the complex. Total cellular doxorubicin fluorescence was similar following CuDox and free doxorubicin incubation. Imaging and mass spectrometry assays indicated that the CuDox complex was initially internalized intact but breaks down over time within cells, with intracellular copper decreasing more rapidly than intracellular doxorubicin. Doxorubicin fluorescence was reduced when complexed with copper, and nuclear fluorescence was reduced when cells were incubated with the CuDox complex as compared with free doxorubicin. Therapeutic efficacy, which typically results from intercalation of doxorubicin with DNA, was equivalent for the CuDox complex and free doxorubicin and was superior to that of liposomal doxorubicin formulations. Taken together, the results suggest that quenched CuDox reaches the nucleus and remains efficacious. In order to design protocols for the use of these temperature-sensitive particles in cancer treatment, the timing of hyperthermia relative to drug administration must be examined. When cells were heated to 42°C prior to the addition of free doxorubicin, nuclear drug accumulation increased by 1.8-fold in cancer cells after 5h, and cytotoxicity increased 1

Exogenous salicylate application affects the lead and copper accumulation characteristics of Lemna gibba L.

Energy Technology Data Exchange (ETDEWEB)

Duman, Fatih; Aksoy, Ahmet; Ozturk, Fatma; Ceylan, Ahmet [Erciyes Univ., Kayseri (Turkey). Dept. of Biology

2010-11-15

Previous studies have shown that salicylates can change the ion permeability of root cells. Therefore the possible effects of exogenous salicylate application on lead (Pb) and copper (Cu) accumulation and its protective role against DNA damage due to metal exposure in Lemna gibba were studied. L. gibba was exposed to 5, 10, and 25 {mu}M Pb and Cu for six days in the presence and absence of sodium salicylate (SA) (0.1, 0.5, and 1 mM). At all concentrations tested, SA application decreased Pb accumulation. On the other hand, application of 0.5 mM SA increased Cu accumulation. SA did not reduce DNA damage resulting from Pb and Cu toxicity. In summary, SA may be useful for reducing Pb accumulation, and application of SA at 0.5 mM may be useful for the phytoextraction of Cu. (orig.)

Long-term soil accumulation of chromium, copper, and arsenic adjacent to preservative-treated wood.

Science.gov (United States)

S. Lebow; D. Foster; J. Evans

2004-01-01

Chromated copper arsenate (CCA) treated wood has been used extensively in outdoor applications. The Environmental Protection Agency (EPA) and CCA producers recently reached an agreement to limit future use of CCA for some types of applications. One area of concern is the long-term accumulation of leached CCA in soil adjacent to treated wood structures. Interpreting...

Intracellular vorinostat accumulation and its relationship to histone deacetylase activity in soft tissue sarcoma patients.

Science.gov (United States)

Burhenne, Jürgen; Liu, Lu; Heilig, Christoph E; Meid, Andreas D; Leisen, Margarete; Schmitt, Thomas; Kasper, Bernd; Haefeli, Walter E; Mikus, Gerd; Egerer, Gerlinde

2017-08-01

In the regulation of chromatin-structure and histone function, histone deacetylases (HDACs) are key enzymes and thus modulators of epigenetic regulation and gene expression. Accesses of the HDAC inhibitor vorinostat to intracellular compartments are essential to exert epigenetic effects. In ten sarcoma patients receiving oral Zolinza (400 mg qd) vorinostat concentrations in plasma and peripheral blood mononuclear cells (PBMCs) were quantified using validated LC/MS/MS assays to determine intracellular and extracellular pharmacokinetic data. Cellular HDAC activity was evaluated using a fluorogenic assay. Concentration-response relationships were established between intracellular and extracellular vorinostat concentrations and HDAC inhibition in PBMCs. Pharmacokinetics of vorinostat and its two main inactive metabolites were determined over 8 h in plasma and PBMCs. Steady state AUCs (±SD) and T 1/2 (±SD) were calculated to 4.61 ± 0.87 h µM and 1.73 ± 0.69 h (plasma) and 15.2 ± 9.03 h µM and 5.30 ± 4.27 h (PBMCs). Intracellular accumulation of vorinostat was determined together with prolonged vorinostat elimination in PBMCs. Cellular HDAC inhibition increased parallel with vorinostat concentrations in plasma and PBMCs. For effective inhibition of cellular HDACs (IC 50 ) vorinostat concentrations of 0.05 µM in plasma and 0.17 µM in PBMCs were necessary. HDAC inhibition closely followed intracellular vorinostat concentrations and was short-lasting, which may contribute to the limited efficacy seen with vorinostat in solid tumors so far.

Metabolic states following accumulation of intracellular aggregates: implications for neurodegenerative diseases.

Directory of Open Access Journals (Sweden)

Alexei Vazquez

Full Text Available The formation of intracellular aggregates is a common etiology of several neurodegenerative diseases. Mitochondrial defects and oxidative stress has been pointed as the major mechanistic links between the accumulation of intracellular aggregates and cell death. In this work we propose a "metabolic cell death by overcrowding" as an alternative hypothesis. Using a model of neuron metabolism, we predict that as the concentration of protein aggregates increases the neurons transit through three different metabolic phases. The first phase (0-6 mM corresponds with the normal neuron state, where the neuronal activity is sustained by the oxidative phosphorylation of lactate. The second phase (6-8.6 mM is characterized by a mixed utilization of lactate and glucose as energy substrates and a switch from ammonia uptake to ammonia release by neurons. In the third phase (8.6-9.3 mM neurons are predicted to support their energy demands from glycolysis and an alternative pathway for energy generation, involving reactions from serine synthesis, one carbon metabolism and the glycine cleavage system. The model also predicts a decrease in the maximum neuronal capacity for energy generation with increasing the concentration of protein aggregates. Ultimately this maximum capacity becomes zero when the protein aggregates reach a concentration of about 9.3 mM, predicting the cessation of neuronal activity.

Effects of exposure to high glucose on primary cultured hippocampal neurons: involvement of intracellular ROS accumulation.

Science.gov (United States)

Liu, Di; Zhang, Hong; Gu, Wenjuan; Zhang, Mengren

2014-06-01

Recent studies showed that hyperglycemia is the main trigger of diabetic cognitive impairment and can cause hippocampus abnormalities. The goal of this study is to explore the effects of different concentrations of high glucose for different exposure time on cell viability as well as intracellular reactive oxygen species (ROS) generation of primary cultured hippocampal neurons. Hippocampal neurons were exposed to different concentrations of high glucose (50, 75, 100, 125, and 150 mM) for 24, 48, 72 and 96 h. Cell viability and nuclear morphology were evaluated by MTT and Hoechst assays, respectively. Intracellular ROS were monitored using the fluorescent probe DCFH-DA. The results showed that, compared with control group, the cell viability of all high glucose-treated groups decreased significantly after 72 h and there also was a significant increase of apoptotic nuclei in high glucose-treated groups from 72 to 96 h. Furthermore, 50 mM glucose induced a peak rise in ROS generation at 24 h and the intracellular ROS levels of 50 mM glucose group were significantly higher than the corresponding control group from 6 to 72 h. These results suggest that hippocampal neurons could be injured by high glucose exposure and the neuronal injury induced by high glucose is potentially mediated through intracellular ROS accumulation.

Bio-accumulation of copper, zinc, iron and manganese in oyster Saccostrea cucullata, Snail Cerithium rubus and Clam Tellina angulata from the Bombay coast

Digital Repository Service at National Institute of Oceanography (India)

Krishnakumari, L.; Nair, V.R; Moraes, C.

accumulation was high in S. cucullata, manganese in C. rubus and iron in T. angulata. Similarly, copper and zinc in S. cucullata and copper in C. rubus were found occasionally higher than accepted health standards...

Erythrocytic ferroportin reduces intracellular iron accumulation, hemolysis, and malaria risk.

Science.gov (United States)

Zhang, De-Liang; Wu, Jian; Shah, Binal N; Greutélaers, Katja C; Ghosh, Manik C; Ollivierre, Hayden; Su, Xin-Zhuan; Thuma, Philip E; Bedu-Addo, George; Mockenhaupt, Frank P; Gordeuk, Victor R; Rouault, Tracey A

2018-03-30

Malaria parasites invade red blood cells (RBCs), consume copious amounts of hemoglobin, and severely disrupt iron regulation in humans. Anemia often accompanies malaria disease; however, iron supplementation therapy inexplicably exacerbates malarial infections. Here we found that the iron exporter ferroportin (FPN) was highly abundant in RBCs, and iron supplementation suppressed its activity. Conditional deletion of the Fpn gene in erythroid cells resulted in accumulation of excess intracellular iron, cellular damage, hemolysis, and increased fatality in malaria-infected mice. In humans, a prevalent FPN mutation, Q248H (glutamine to histidine at position 248), prevented hepcidin-induced degradation of FPN and protected against severe malaria disease. FPN Q248H appears to have been positively selected in African populations in response to the impact of malaria disease. Thus, FPN protects RBCs against oxidative stress and malaria infection. Copyright © 2018 The Authors, some rights reserved; exclusive licensee American Association for the Advancement of Science. No claim to original U.S. Government Works.

A horizontally gene transferred copper resistance locus confers hyper‐resistance to antibacterial copper toxicity and enables survival of community acquired methicillin resistant Staphylococcus aureus USA300 in macrophages

Science.gov (United States)

Purves, Joanne; Thomas, Jamie; Riboldi, Gustavo P.; Zapotoczna, Marta; Tarrant, Emma; Andrew, Peter W.; Londoño, Alejandra; Planet, Paul J.; Geoghegan, Joan A.; Waldron, Kevin J.

2018-01-01

Summary Excess copper is highly toxic and forms part of the host innate immune system's antibacterial arsenal, accumulating at sites of infection and acting within macrophages to kill engulfed pathogens. We show for the first time that a novel, horizontally gene transferred copper resistance locus (copXL), uniquely associated with the SCCmec elements of the highly virulent, epidemic, community acquired methicillin resistant Staphylococcus aureus (CA‐MRSA) USA300, confers copper hyper‐resistance. These genes are additional to existing core genome copper resistance mechanisms, and are not found in typical S. aureus lineages, but are increasingly identified in emerging pathogenic isolates. Our data show that CopX, a putative P1B‐3‐ATPase efflux transporter, and CopL, a novel lipoprotein, confer copper hyper‐resistance compared to typical S. aureus strains. The copXL genes form an operon that is tightly repressed in low copper environments by the copper regulator CsoR. Significantly, CopX and CopL are important for S. aureus USA300 intracellular survival within macrophages. Therefore, the emergence of new S. aureus clones with the copXL locus has significant implications for public health because these genes confer increased resistance to antibacterial copper toxicity, enhancing bacterial fitness by altering S. aureus interaction with innate immunity. PMID:29521441

Variation in copper and zinc tolerance and accumulation in 12 willow clones: implications for phytoextraction*

Science.gov (United States)

Yang, Wei-dong; Wang, Yu-yan; Zhao, Feng-liang; Ding, Zhe-li; Zhang, Xin-cheng; Zhu, Zhi-qiang; Yang, Xiao-e

2014-01-01

Willows (Salix spp.) have shown high potential for the phytoextraction of heavy metals. This study compares variations in copper (Cu) and zinc (Zn) tolerance and accumulation potential among 12 willow clones grown in a nutrient solution treated with 50 μmol/L of Cu or Zn, respectively. The results showed differences in the tolerance and accumulation of Cu and Zn with respect to different species/clones. The biomass variation among clones in response to Cu or Zn exposure ranged from the stimulation of growth to inhibition, and all of the clones tested showed higher tolerance to Cu than to Zn. The clones exhibited less variation in Cu accumulation but larger variation in Zn accumulation. Based on translocation factors, it was found that most of the Cu was retained in the roots and that Zn was more mobile than Cu for all clones. It is concluded that most willow clones are good accumulators of Zn and Cu. PMID:25183033

Increased extracellular and intracellular Ca{sup 2+} lead to adipocyte accumulation in bone marrow stromal cells by different mechanisms

Energy Technology Data Exchange (ETDEWEB)

Hashimoto, Ryota, E-mail: hryota@juntendo.ac.jp [Department of Physiology, Juntendo University Faculty of Medicine, Hongo 2-1-1, Bunkyo-ku, Tokyo 113-8421 (Japan); Katoh, Youichi, E-mail: katoyo@juntendo-urayasu.jp [Juntendo University Faculty of International Liberal Arts, Hongo 2-1-1, Bunkyo-ku, Tokyo 113-8421 (Japan); Department of Cardiology, Juntendo University Faculty of Medicine, Hongo 2-1-1, Bunkyo-ku, Tokyo 113-8421 (Japan); Miyamoto, Yuki [Juntendo University Faculty of Health Care and Nursing, Takasu 2-5-1, Urayasu-shi, Chiba 279-0023 (Japan); Itoh, Seigo; Daida, Hiroyuki [Department of Cardiology, Juntendo University Faculty of Medicine, Hongo 2-1-1, Bunkyo-ku, Tokyo 113-8421 (Japan); Nakazato, Yuji [Center for Environmental Research, Department of Cardiology, Juntendo University Faculty of Medicine Urayasu Hospital, Tomioka 2-1-1, Urayasu-shi, Chiba 279-0022 (Japan); Okada, Takao [Department of Physiology, Juntendo University Faculty of Medicine, Hongo 2-1-1, Bunkyo-ku, Tokyo 113-8421 (Japan)

2015-02-20

Mesenchymal stem cells found in bone marrow stromal cells (BMSCs) are the common progenitors for both adipocyte and osteoblast. An increase in marrow adipogenesis is associated with age-related osteopenia and anemia. Both extracellular and intracellular Ca{sup 2+} ([Ca{sup 2+}]{sub o} and [Ca{sup 2+}]{sub i}) are versatile signaling molecules that are involved in the regulation of cell functions, including proliferation and differentiation. We have recently reported that upon treatment of BMSCs with insulin and dexamethasone, both high [Ca{sup 2+}]{sub o} and high [Ca{sup 2+}]{sub i} enhanced adipocyte accumulation, which suggested that increases in [Ca{sup 2+}]{sub o} caused by bone resorption may accelerate adipocyte accumulation in aging and diabetic patients. In this study, we used primary mouse BMSCs to investigate the mechanisms by which high [Ca{sup 2+}]{sub o} and high [Ca{sup 2+}]{sub i} may enhance adipocyte accumulation. In the process of adipocyte accumulation, two important keys are adipocyte differentiation and the proliferation of BMSCs, which have the potential to differentiate into adipocytes. Use of MTT assay and real-time RT-PCR revealed that high [Ca{sup 2+}]{sub i} (ionomycin)-dependent adipocyte accumulation is caused by enhanced proliferation of BMSCs but not enhanced differentiation into adipocytes. Using fura-2 fluorescence-based approaches, we showed that high [Ca{sup 2+}]{sub o} (addition of CaCl{sub 2}) leads to increases in [Ca{sup 2+}]{sub i}. Flow cytometric methods revealed that high [Ca{sup 2+}]{sub o} suppressed the phosphorylation of ERK independently of intracellular Ca{sup 2+}. The inhibition of ERK by U0126 and PD0325901 enhanced the differentiation of BMSCs into adipocytes. These data suggest that increased extracellular Ca{sup 2+} provides the differentiation of BMSCs into adipocytes by the suppression of ERK activity independently of increased intracellular Ca{sup 2+}, which results in BMSC proliferation. - Highlights: �

Rate and Regulation of Copper Transport by Human Copper Transporter 1 (hCTR1)*

Science.gov (United States)

Maryon, Edward B.; Molloy, Shannon A.; Ivy, Kristin; Yu, Huijun; Kaplan, Jack H.

2013-01-01

Human copper transporter 1 (hCTR1) is a homotrimer of a 190-amino acid monomer having three transmembrane domains believed to form a pore for copper permeation through the plasma membrane. The hCTR1-mediated copper transport mechanism is not well understood, nor has any measurement been made of the rate at which copper ions are transported by hCTR1. In this study, we estimated the rate of copper transport by the hCTR1 trimer in cultured cells using 64Cu uptake assays and quantification of plasma membrane hCTR1. For endogenous hCTR1, we estimated a turnover number of about 10 ions/trimer/s. When overexpressed in HEK293 cells, a second transmembrane domain mutant of hCTR1 (H139R) had a 3-fold higher Km value and a 4-fold higher turnover number than WT. Truncations of the intracellular C-terminal tail and an AAA substitution of the putative metal-binding HCH C-terminal tripeptide (thought to be required for transport) also exhibited elevated transport rates and Km values when compared with WT hCTR1. Unlike WT hCTR1, H139R and the C-terminal mutants did not undergo regulatory endocytosis in elevated copper. hCTR1 mutants combining methionine substitutions that block transport (M150L,M154L) on the extracellular side of the pore and the high transport H139R or AAA intracellular side mutations exhibited the blocked transport of M150L,M154L, confirming that Cu+ first interacts with the methionines during permeation. Our results show that hCTR1 elements on the intracellular side of the hCTR1 pore, including the carboxyl tail, are not essential for permeation, but serve to regulate the rate of copper entry. PMID:23658018

Cross talk between increased intracellular zinc (Zn2+) and accumulation of reactive oxygen species in chemical ischemia.

Science.gov (United States)

Slepchenko, Kira G; Lu, Qiping; Li, Yang V

2017-10-01

Both zinc (Zn 2+ ) and reactive oxygen species (ROS) have been shown to accumulate during hypoxic-ischemic stress and play important roles in pathological processes. To understand the cross talk between the two of them, here we studied Zn 2+ and ROS accumulation by employing fluorescent probes in HeLa cells to further the understanding of the cause and effect relationship of these two important cellular signaling systems during chemical-ischemia, stimulated by oxygen and glucose deprivation (OGD). We observed two Zn 2+ rises that were divided into four phases in the course of 30 min of OGD. The first Zn 2+ rise was a transient, which was followed by a latent phase during which Zn 2+ levels recovered; however, levels remained above a basal level in most cells. The final phase was the second Zn 2+ rise, which reached a sustained plateau called Zn 2+ overload. Zn 2+ rises were not observed when Zn 2+ was removed by TPEN (a Zn 2+ chelator) or thapsigargin (depleting Zn 2+ from intracellular stores) treatment, indicating that Zn 2+ was from intracellular storage. Damaging mitochondria with FCCP significantly reduced the second Zn 2+ rise, indicating that the mitochondrial Zn 2+ accumulation contributes to Zn 2+ overload. We also detected two OGD-induced ROS rises. Two Zn 2+ rises preceded two ROS rises. Removal of Zn 2+ reduced or delayed OGD- and FCCP-induced ROS generation, indicating that Zn 2+ contributes to mitochondrial ROS generation. There was a Zn 2+ -induced increase in the functional component of NADPH oxidase, p47 phox , thus suggesting that NADPH oxidase may mediate Zn 2+ -induced ROS accumulation. We suggest a new mechanism of cross talk between Zn 2+ and mitochondrial ROS through positive feedback processes that eventually causes excessive free Zn 2+ and ROS accumulations during the course of ischemic stress. Copyright © 2017 the American Physiological Society.

Canine Copper-Associated Hepatitis

NARCIS (Netherlands)

Dirksen, Karen; Fieten, Hille

2017-01-01

Copper-associated hepatitis is recognized with increasing frequency in dogs. The disease is characterized by centrolobular hepatic copper accumulation, leading to hepatitis and eventually cirrhosis. The only way to establish the diagnosis is by histologic assessment of copper distribution and copper

Copper changes the yield and cadmium/zinc accumulation and cellular distribution in the cadmium/zinc hyperaccumulator Sedum plumbizincicola

Energy Technology Data Exchange (ETDEWEB)

Li, Zhu [Key Laboratory of Soil Environment and Pollution Remediation, Institute of Soil Science, Chinese Academy of Sciences, Nanjing 210008 (China); University of the Chinese Academy of Sciences, Beijing 100049 (China); Wu, Longhua, E-mail: lhwu@issas.ac.cn [Key Laboratory of Soil Environment and Pollution Remediation, Institute of Soil Science, Chinese Academy of Sciences, Nanjing 210008 (China); Hu, Pengjie [Key Laboratory of Soil Environment and Pollution Remediation, Institute of Soil Science, Chinese Academy of Sciences, Nanjing 210008 (China); Luo, Yongming [Key Laboratory of Soil Environment and Pollution Remediation, Institute of Soil Science, Chinese Academy of Sciences, Nanjing 210008 (China); Yantai Institute of Coastal Zone Research, Yantai 264003 (China); Christie, Peter [Agri-Environment Branch, Agri-Food and Biosciences Institute, Newforge Lane, Belfast BT9 5PX (United Kingdom)

2013-10-15

Highlights: • Low Cu has no significant effect on Sedum plumbizincicola plant growth and Cd and Zn uptake. • Plant held Cu in unactive areas and insoluble forms as de-toxification mechanisms. • Influence of Cu on Zn and Cd uptake and translocation were different. • Cu accumulation in leaf veins may restrain Cd/Zn unloading to the leaves -- Abstract: Non-accumulated metals in mixed metal contaminated soils may affect hyperaccumulator growth and metal accumulation and thus remediation efficiency. Two hydroponics experiments were conducted to investigate the effects of copper (Cu) on cadmium (Cd) and zinc (Zn) accumulation by the Cd/Zn hyperaccumulator Sedum plumbizincicola, Cu toxicity and plant detoxification using chemical sequential extraction of metals, sub-cellular separation, micro synchrotron radiation based X-ray fluorescence, and transmission electron microscopy. Compared with the control (0.31 μM Cu), 5–50 μM Cu had no significant effect on Cd/Zn accumulation, but Cu at 200 μM induced root cell plasmolysis and disordered chloroplast structure. The plants held Cu in the roots and cell walls and complexed Cu in insoluble forms as their main detoxification mechanisms. Exposure to 200 μM Cu for 4 days inhibited plant Cd uptake and translocation but did not affect Zn concentrations in roots and stems. Moreover, unloading of Cd and Zn from stem to leaf was restrained compared to control plants, perhaps due to Cu accumulation in leaf veins. Copper may thus interfere with root Cd uptake and restrain Cd/Zn unloading to the leaves. Further investigation of how Cu affects plant metal uptake may help elucidate the Cd/Zn hyper-accumulating mechanisms of S. plumbizincicola.

Copper changes the yield and cadmium/zinc accumulation and cellular distribution in the cadmium/zinc hyperaccumulator Sedum plumbizincicola

International Nuclear Information System (INIS)

Li, Zhu; Wu, Longhua; Hu, Pengjie; Luo, Yongming; Christie, Peter

2013-01-01

Highlights: • Low Cu has no significant effect on Sedum plumbizincicola plant growth and Cd and Zn uptake. • Plant held Cu in unactive areas and insoluble forms as de-toxification mechanisms. • Influence of Cu on Zn and Cd uptake and translocation were different. • Cu accumulation in leaf veins may restrain Cd/Zn unloading to the leaves -- Abstract: Non-accumulated metals in mixed metal contaminated soils may affect hyperaccumulator growth and metal accumulation and thus remediation efficiency. Two hydroponics experiments were conducted to investigate the effects of copper (Cu) on cadmium (Cd) and zinc (Zn) accumulation by the Cd/Zn hyperaccumulator Sedum plumbizincicola, Cu toxicity and plant detoxification using chemical sequential extraction of metals, sub-cellular separation, micro synchrotron radiation based X-ray fluorescence, and transmission electron microscopy. Compared with the control (0.31 μM Cu), 5–50 μM Cu had no significant effect on Cd/Zn accumulation, but Cu at 200 μM induced root cell plasmolysis and disordered chloroplast structure. The plants held Cu in the roots and cell walls and complexed Cu in insoluble forms as their main detoxification mechanisms. Exposure to 200 μM Cu for 4 days inhibited plant Cd uptake and translocation but did not affect Zn concentrations in roots and stems. Moreover, unloading of Cd and Zn from stem to leaf was restrained compared to control plants, perhaps due to Cu accumulation in leaf veins. Copper may thus interfere with root Cd uptake and restrain Cd/Zn unloading to the leaves. Further investigation of how Cu affects plant metal uptake may help elucidate the Cd/Zn hyper-accumulating mechanisms of S. plumbizincicola

Comparative effects of dissolved copper and copper oxide nanoparticle exposure to the sea anemone, Exaiptasia pallida

Energy Technology Data Exchange (ETDEWEB)

Siddiqui, Samreen; Goddard, Russell H.; Bielmyer-Fraser, Gretchen K., E-mail: gkbielmyer@valdosta.edu

2015-03-15

Highlights: • Differences between CuO NP and CuCl{sub 2} exposure were characterized. • Copper accumulation in E. pallida was concentration-dependent. • E. pallida exposed to CuCl{sub 2} accumulated higher copper tissue burdens. • The oxidative stress response was greater in E. pallida exposed to CuO NP. • Both forms of copper inhibited CA activity in E. pallida. - Abstract: Increasing use of metal oxide nanoparticles (NP) by various industries has resulted in substantial output of these NP into aquatic systems. At elevated concentrations, NP may interact with and potentially affect aquatic organisms. Environmental implications of increased NP use are largely unknown, particularly in marine systems. This research investigated and compared the effects of copper oxide (CuO) NP and dissolved copper, as copper chloride (CuCl{sub 2}), on the sea anemone, Exaiptasia pallida. Sea anemones were collected over 21 days and tissue copper accumulation and activities of the enzymes: catalase, glutathione peroxidase, glutathione reductase, and carbonic anhydrase were quantified. The size and shape of CuO NP were observed using a ecanning electron microscope (SEM) and the presence of copper was confirmed by using Oxford energy dispersive spectroscopy systems (EDS/EDX). E. pallida accumulated copper in their tissues in a concentration- and time-dependent manner, with the animals exposed to CuCl{sub 2} accumulating higher tissue copper burdens than those exposed to CuO NP. As a consequence of increased copper exposure, as CuO NP or CuCl{sub 2}, anemones increased activities of all of the antioxidant enzymes measured to some degree, and decreased the activity of carbonic anhydrase. Anemones exposed to CuO NP generally had higher anti-oxidant enzyme activities than those exposed to the same concentrations of CuCl{sub 2}. This study is useful in discerning differences between CuO NP and dissolved copper exposure and the findings have implications for exposure of aquatic

Cellular copper homeostasis: current concepts on its interplay with glutathione homeostasis and its implication in physiology and human diseases.

Science.gov (United States)

Bhattacharjee, Ashima; Chakraborty, Kaustav; Shukla, Aditya

2017-10-18

Copper is a trace element essential for almost all living organisms. But the level of intracellular copper needs to be tightly regulated. Dysregulation of cellular copper homeostasis leading to various diseases demonstrates the importance of this tight regulation. Copper homeostasis is regulated not only within the cell but also within individual intracellular compartments. Inactivation of export machinery results in excess copper being redistributed into various intracellular organelles. Recent evidence suggests the involvement of glutathione in playing an important role in regulating copper entry and intracellular copper homeostasis. Therefore interplay of both homeostases might play an important role within the cell. Similar to copper, glutathione balance is tightly regulated within individual cellular compartments. This review explores the existing literature on the role of glutathione in regulating cellular copper homeostasis. On the one hand, interplay of glutathione and copper homeostasis performs an important role in normal physiological processes, for example neuronal differentiation. On the other hand, perturbation of the interplay might play a key role in the pathogenesis of copper homeostasis disorders.

Intermittent Hypoxia Inhibits Na+-H+ Exchange-Mediated Acid Extrusion Via Intracellular Na+ Accumulation in Cardiomyocytes

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Huai-Ren Chang

2018-04-01

Full Text Available Background/Aims: Intermittent hypoxia (IH has been shown to exert preconditioning-like cardioprotective effects. It also has been reported that IH preserves intracellular pH (pHi during ischemia and protects cardiomyocytes against ischemic reperfusion injury. However, the exact mechanism is still unclear. Methods: In this study, we used proton indicator BCECF-AM to analyze the rate of pHi recovery from acidosis in the IH model of rat neonatal cardiomyocytes. Neonatal cardiomyocytes were first treated with repetitive hypoxia-normoxia cycles for 1-4 days. Cells were then acid loaded with NH4Cl, and the rate of pHi recovery from acidosis was measured. Results: We found that the pHi recovery rate from acidosis was much slower in the IH group than in the room air (RA group. When we treated cardiomyocytes with Na+-H+ exchange (NHE inhibitors (Amiloride and HOE642 or Na+-free Tyrode solution during the recovery, there was no difference between RA and IH groups. We also found intracellular Na+ concentration ([Na+]i significantly increased after IH exposure for 4 days. However, the phenomenon could be abolished by pretreatment with ROS inhibitors (SOD and phenanathroline, intracellular calcium chelator or Na+-Ca2+ exchange (NCX inhibitor. Furthermore, the pHi recovery rate from acidosis became faster in the IH group than in the RA group when inhibition of NCX activity. Conclusions: These results suggest that IH would induce the elevation of ROS production. ROS then activates Ca2+-efflux mode of NCX and results in intracellular Na+ accumulation. The rise of [Na+]i further inhibits the activity of NHE-mediated acid extrusion and retards the rate of pHi recovery from acidosis during IH.

Intracellular biosynthesis and removal of copper nanoparticles by dead biomass of yeast isolated from the wastewater of a mine in the Brazilian Amazonia.

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Marcia R Salvadori

Full Text Available In this study was developed a natural process using a biological system for the biosynthesis of nanoparticles (NPs and possible removal of copper from wastewater by dead biomass of the yeast Rhodotorula mucilaginosa. Dead and live biomass of Rhodotorula mucilaginosa was used to analyze the equilibrium and kinetics of copper biosorption by the yeast in function of the initial metal concentration, contact time, pH, temperature, agitation and inoculum volume. Dead biomass exhibited the highest biosorption capacity of copper, 26.2 mg g(-1, which was achieved within 60 min of contact, at pH 5.0, temperature of 30°C, and agitation speed of 150 rpm. The equilibrium data were best described by the Langmuir isotherm and Kinetic analysis indicated a pseudo-second-order model. The average size, morphology and location of NPs biosynthesized by the yeast were determined by scanning electron microscopy (SEM, energy dispersive X-ray spectroscopy (EDS and transmission electron microscopy (TEM. The shape of the intracellularly synthesized NPs was mainly spherical, with an average size of 10.5 nm. The X-ray photoelectron spectroscopy (XPS analysis of the copper NPs confirmed the formation of metallic copper. The dead biomass of Rhodotorula mucilaginosa may be considered an efficiently bioprocess, being fast and low-cost to production of copper nanoparticles and also a probably nano-adsorbent of this metal ion in wastewater in bioremediation process.

Jinlida granule inhibits palmitic acid induced-intracellular lipid accumulation and enhances autophagy in NIT-1 pancreatic β cells through AMPK activation.

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Wang, Dingkun; Tian, Min; Qi, Yuan; Chen, Guang; Xu, Lijun; Zou, Xin; Wang, Kaifu; Dong, Hui; Lu, Fuer

2015-02-23

Jinlida granule (JLDG), composed of seventeen Chinese medical herbs, is a widely used Chinese herbal prescription for treating diabetes mellitus. However, the mechanism underlying this effect remains unclear. To determine the main components in JLDG and to explore the effect of JLDG on autophagy and lipid accumulation in NIT-1 pancreatic β cells exposed to politic acid (PA) through AMP activated protein kinase (AMPK) signaling pathway. JLDG was prepared and the main components contained in the granules were identified by ultra performance liquid chromatography (UPLC) fingerprint. Intracellular lipid accumulation in NIT-1 cells was induced by culturing with medium containing PA. Intracellular lipid droplets were observed by Oil Red O staining and triglyceride (TG) content was measured by colorimetric assay. The formation of autophagosomes was observed under transmission electron microscope. The expression of AMPK and phospho-AMPK (pAMPK) proteins as well as its downstream fatty acid metabolism-related proteins (fatty acid synthase, FAS; acetyl-coA carboxylase, ACC; carnitine acyltransferase 1, CPT-1) and autophagy-related genes (mammal target of rapamycin, mTOR; tuberous sclerosis complex 1, TSC1; microtubule-associated protein 1 light chain 3, LC3-II) were determined by Western blot. The expression of sterol regulating element binding protein 1c (SREBP-1c) mRNA was examined by real time PCR (RT-PCR). Our data showed that JLDG could significantly reduce PA-induced intracellular lipid accumulation in NIT-1 pancreatic β cells. This effect was associated with increased protein expression of pAMPK and AMPK in NIT-1 cells. Treatment with JLDG also decreased the expression of AMPK downstream lipogenic genes (SREBP-1c mRNA, FAS and ACC proteins) whereas increased the expression of fatty acid oxidation gene (CPT-1 protein). Additionally, JLDG-treated cells displayed a markedly increase in the number of autophagosomes which was accompanied by the down-regulation of m

Trigeminal ganglion neurons of mice show intracellular chloride accumulation and chloride-dependent amplification of capsaicin-induced responses.

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Nicole Schöbel

Full Text Available Intracellular Cl(- concentrations ([Cl(-](i of sensory neurons regulate signal transmission and signal amplification. In dorsal root ganglion (DRG and olfactory sensory neurons (OSNs, Cl(- is accumulated by the Na(+-K(+-2Cl(- cotransporter 1 (NKCC1, resulting in a [Cl(-](i above electrochemical equilibrium and a depolarizing Cl(- efflux upon Cl(- channel opening. Here, we investigate the [Cl(-](i and function of Cl(- in primary sensory neurons of trigeminal ganglia (TG of wild type (WT and NKCC1(-/- mice using pharmacological and imaging approaches, patch-clamping, as well as behavioral testing. The [Cl(-](i of WT TG neurons indicated active NKCC1-dependent Cl(- accumulation. Gamma-aminobutyric acid (GABA(A receptor activation induced a reduction of [Cl(-](i as well as Ca(2+ transients in a corresponding fraction of TG neurons. Ca(2+ transients were sensitive to inhibition of NKCC1 and voltage-gated Ca(2+ channels (VGCCs. Ca(2+ responses induced by capsaicin, a prototypical stimulus of transient receptor potential vanilloid subfamily member-1 (TRPV1 were diminished in NKCC1(-/- TG neurons, but elevated under conditions of a lowered [Cl(-](o suggesting a Cl(--dependent amplification of capsaicin-induced responses. Using next generation sequencing (NGS, we found expression of different Ca(2+-activated Cl(- channels (CaCCs in TGs of mice. Pharmacological inhibition of CaCCs reduced the amplitude of capsaicin-induced responses of TG neurons in Ca(2+ imaging and electrophysiological recordings. In a behavioral paradigm, NKCC1(-/- mice showed less avoidance of the aversive stimulus capsaicin. In summary, our results strongly argue for a Ca(2+-activated Cl(--dependent signal amplification mechanism in TG neurons that requires intracellular Cl(- accumulation by NKCC1 and the activation of CaCCs.

An atmospheric-pressure cold plasma leads to apoptosis in Saccharomyces cerevisiae by accumulating intracellular reactive oxygen species and calcium

International Nuclear Information System (INIS)

Ma, R N; Zhang, Q; Tian, Y; Su, B; Zhang, J; Fang, J; Feng, H Q; Liang, Y D

2013-01-01

A non-thermal plasma is known to induce apoptosis of various cells but the mechanism is not yet clear. A eukaryotic model organism Saccharomyces cerevisiaewas used to investigate the cellular and biochemical regulations of cell apoptosis and cell cycle after an atmospheric-pressure cold plasma treatment. More importantly, intracellular calcium (Ca 2+ ) was first involved in monitoring the process of plasma-induced apoptosis in this study. We analysed the cell apoptosis and cell cycle by flow cytometry and observed the changes in intracellular reactive oxygen species (ROS) and Ca 2+ concentration, cell mitochondrial membrane potential (Δψ m ) as well as nuclear DNA morphology via fluorescence staining assay. All experimental results indicated that plasma-generated ROS leads to the accumulation of intracellular ROS and Ca 2+ that ultimately contribute to apoptosis associated with cell cycle arrest at G1 phase through depolarization of Δψ m and fragmenting nuclear DNA. This work provides a novel insight into the physical and biological mechanism of apoptosis induced by a plasma which could benefit for promoting the development of plasmas applied to cancer therapy. (paper)

The Menkes and Wilson disease genes counteract in copper toxicosis in Labrador retrievers: a new canine model for copper-metabolism disorders

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Hille Fieten

2016-01-01

Full Text Available The deleterious effects of a disrupted copper metabolism are illustrated by hereditary diseases caused by mutations in the genes coding for the copper transporters ATP7A and ATP7B. Menkes disease, involving ATP7A, is a fatal neurodegenerative disorder of copper deficiency. Mutations in ATP7B lead to Wilson disease, which is characterized by a predominantly hepatic copper accumulation. The low incidence and the phenotypic variability of human copper toxicosis hamper identification of causal genes or modifier genes involved in the disease pathogenesis. The Labrador retriever was recently characterized as a new canine model for copper toxicosis. Purebred dogs have reduced genetic variability, which facilitates identification of genes involved in complex heritable traits that might influence phenotype in both humans and dogs. We performed a genome-wide association study in 235 Labrador retrievers and identified two chromosome regions containing ATP7A and ATP7B that were associated with variation in hepatic copper levels. DNA sequence analysis identified missense mutations in each gene. The amino acid substitution ATP7B:p.Arg1453Gln was associated with copper accumulation, whereas the amino acid substitution ATP7A:p.Thr327Ile partly protected against copper accumulation. Confocal microscopy indicated that aberrant copper metabolism upon expression of the ATP7B variant occurred because of mis-localization of the protein in the endoplasmic reticulum. Dermal fibroblasts derived from ATP7A:p.Thr327Ile dogs showed copper accumulation and delayed excretion. We identified the Labrador retriever as the first natural, non-rodent model for ATP7B-associated copper toxicosis. Attenuation of copper accumulation by the ATP7A mutation sheds an interesting light on the interplay of copper transporters in body copper homeostasis and warrants a thorough investigation of ATP7A as a modifier gene in copper-metabolism disorders. The identification of two new functional

Resistance of Streptococcus bovis to acetic acid at low pH: Relationship between intracellular pH and anion accumulation

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Russell, J.B. (Cornell Univ., Ithaca, NY (USA))

1991-01-01

Streptococcus bovis JB1, an acid-tolerant ruminal bacterium, was able to grown at pHs from 6.7 to 4.5, and 100 mM acetate had little effect on growth rate or proton motive force across the cell membrane. When S. bovis was grown in glucose-limited chemostats at pH 5.2, the addition of sodium acetate (as much as 100 mM) had little effect on the production of bacterial protein. At higher concentrations of sodium acetate (100 to 360 mM), production of bacterial protein declined, but this decrease could largely be explained by a shift in fermentation products (acetate, formate, and ethanol production to lactate production) and a decline in ATP production (3 ATP per glucose versus 2 ATP per glucose). Y{sub ATP} (grams of cells per mole at ATP) was not decreased significantly even by high concentrations of acetate. Cultures supplemented with 100 mM sodium acetate took up ({sup 14}C)acetate and ({sup 14}C)benzoate in accordance with the Henderson-Hasselbalch equation and gave similar estimates of intracellular pH. As the extracellular pH declined, S. bovis allowed its intracellular pH to decrease and maintained a relatively constant pH gradient across the cell membrane (0.9 unit). The decrease in intracellular pH prevented S. bovis from accumulating large amounts of acetate anion. On the basis of these results it did not appear that acetate was acting as an uncoupler. The sensitivity of other bacteria to volatile fatty acids at low pH is explained most easily by a high transmembrane pH gradient and anion accumulation.

Resistance of Streptococcus bovis to acetic acid at low pH: Relationship between intracellular pH and anion accumulation

International Nuclear Information System (INIS)

Russell, J.B.

1991-01-01

Streptococcus bovis JB1, an acid-tolerant ruminal bacterium, was able to grown at pHs from 6.7 to 4.5, and 100 mM acetate had little effect on growth rate or proton motive force across the cell membrane. When S. bovis was grown in glucose-limited chemostats at pH 5.2, the addition of sodium acetate (as much as 100 mM) had little effect on the production of bacterial protein. At higher concentrations of sodium acetate (100 to 360 mM), production of bacterial protein declined, but this decrease could largely be explained by a shift in fermentation products (acetate, formate, and ethanol production to lactate production) and a decline in ATP production (3 ATP per glucose versus 2 ATP per glucose). Y ATP (grams of cells per mole at ATP) was not decreased significantly even by high concentrations of acetate. Cultures supplemented with 100 mM sodium acetate took up [ 14 C]acetate and [ 14 C]benzoate in accordance with the Henderson-Hasselbalch equation and gave similar estimates of intracellular pH. As the extracellular pH declined, S. bovis allowed its intracellular pH to decrease and maintained a relatively constant pH gradient across the cell membrane (0.9 unit). The decrease in intracellular pH prevented S. bovis from accumulating large amounts of acetate anion. On the basis of these results it did not appear that acetate was acting as an uncoupler. The sensitivity of other bacteria to volatile fatty acids at low pH is explained most easily by a high transmembrane pH gradient and anion accumulation

Influence of sludge retention time on tolerance of copper toxicity for polyphosphate accumulating organisms linked to polyhydroxyalkanoates metabolism and phosphate removal.

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Tsai, Yung-Pin; Chen, Hsiu-Ting

2011-12-01

This study explored the influence of sludge retention time (SRT) on tolerance of copper invasion for polyphosphate accumulating organisms (PAOs) in an enhanced biological phosphorus removal (EBPR). The experimental data showed the anaerobic polyhydroxyalkanoates (PHA) storage for the sludge at 10d SRT was less influenced by copper invasion than those at 5d and 15d SRTs. The reaction of PAOs aerobically taking up phosphate for the sludge at 5d or 15d SRT almost ceased at 2 mg Cu L(-1), whereas PAOs in the sludge at 10d SRT retained half of the ability to take up phosphate. Both the PHAs degradation and synthesis rates decreased with increasing copper concentration, regardless of the SRTs. However, the copper inhibition of the former was greater than that of the later. Copyright © 2011 Elsevier Ltd. All rights reserved.

The copper-transporting ATPase pump and its potential role in copper-tolerance

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Katie Ohno; C.A. Clausen; Frederick Green; G. Stanosz

2016-01-01

Copper-tolerant brown-rot decay fungi exploit intricate mechanisms to neutralize the efficacy of copper-containing preservative formulations. The production and accumulation oxalate is the most widely recognized theory regarding the mechanism of copper-tolerance in these fungi. The role of oxalate, however, may be only one part of a series of necessary components...

APPLICATION OF METAL RESISTANT BACTERIA BY MUTATIONAL ENHANCMENT TECHNIQUE FOR BIOREMEDIATION OF COPPER AND ZINC FROM INDUSTRIAL WASTES

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M. R. Shakibaie ، A. Khosravan ، A. Frahmand ، S. Zare

2008-10-01

Full Text Available In this research, using mutation in the metal resistant bacteria, the bioremediation of the copper and zinc from copper factory effluents was investigated. Wastewater effluents from flocculation and rolling mill sections of a factory in the city of Kerman were collected and used for further experiments. 20 strains of Pseudomonas spp. were isolated from soil and effluents surrounding factory and identified by microbiological methods. Minimum inhibitory concentrations for copper (Cu and zinc (Zn were determined by agar dilution method. Those strains that exhibited highest minimum inhibitory concentrations values to the metals (5mM were subjected to 400-3200 mg/L concentrations of the three mutagenic agents, acriflavine, acridine orange and ethidium bromide. After determination of subinhibitory concentrations, the minimum inhibitory concentrations values for copper and zinc metal ions were again determined, which showed more than 10 fold increase in minimum inhibitory concentrations value (10 mM for Cu and 20 mM for Zn with P≤0.05. The atomic absorption spectroscopy of dried biomass obtained from resistant strains after exposure to mutagenic agents revealed that strains 13 accumulate the highest amount of intracellular copper (0.35% Cu/mg dried biomass and strain 10 showed highest accumulation of zinc (0.3% Zn/mg dried biomass respectively with P≤0.05. From above results it was concluded that the treatment of industrial waste containing heavy metals by artificially mutated bacteria may be appropriate solution for effluent disposal problems.

Iodinated chlorin p6 copper complex induces anti-proliferative effect in oral cancer cells through elevation of intracellular reactive oxygen species.

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Sarbadhikary, Paromita; Dube, Alok

2017-11-01

We investigated the anticancer chemotoxicity of previously reported iodinated chlorin p 6 copper complex (ICp 6 -Cu), a novel chlorophyll derivative in which copper is attached to the side chain carboxylate groups via coordination. Human oral carcinoma cells NT8e, 4451 and the non-cancerous keratinocyte HaCaT cells were treated with ICp 6 -Cu for 48 h in dark and cell viability, proliferation and morphological alterations were examined. ICp 6 -Cu showed pronounced cytotoxicity in cancer cells with IC 50 ∼40 μM, whereas, the viability of HaCaT cells was not affected. Cell proliferation assay revealed that ICp 6 -Cu at IC 50 concentration led to complete inhibition of cell proliferation in both the cell lines. Cell morphology studied by confocal microscopy showed absence of cell death via necrosis or apoptosis. Instead, the treated cells displayed distinct features of non-apoptotic death such as highly vacuolated cytoplasm, lysosomal membrane permeabilization and damage to cytoskeleton F-actin filaments. In addition, ICp 6 -Cu treatment led to time dependent increase in the intracellular level of reactive oxygen species (ROS) and the cytotoxicity of ICp 6 -Cu was significantly inhibited by pre-treatment of cells with antioxidants (glutathione and trolox). These findings revealed that ICp 6 -Cu is a potent chemotoxic agent which can induce cytotoxic effect in cancer cells through elevation of intracellular ROS. It is suggested that ICp 6 -Cu may provide tumor selective chemotoxicity by exploiting difference of redox environment in normal and cancer cells. Copyright © 2017 Elsevier B.V. All rights reserved.

Oxalic acid overproduction by copper-tolerant brown-rot basidiomycetes on southern yellow pine treated with copper-based preservatives

Science.gov (United States)

Carol A. Clausen; Frederick Green

2003-01-01

Accumulation of oxalic acid (OA) by brown-rot fungi and precipitation of copper oxalate crystals in wood decayed by copper-tolerant decay fungi has implicated OA in the mechanism of copper tolerance. Understanding the role of OA in copper tolerance is important due to an increasing reliance on copper-based wood preservatives. In this study, four copper-tolerant brown-...

Copper use and accumulation in catfish culture in the Mekong Delta, Vietnam.

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Marcussen, Helle; Løjmand, Helle; Dalsgaard, Anders; Hai, Dao M; Holm, Peter E

2014-01-01

Aquaculture of Pangasius hypophthalmus (striped catfish) in Vietnam reached 1.1 million tonnes in 2011 and catfish fillets are exported worldwide. The intensive cultures of catfish mainly in earth ponds have made it necessary to apply CuSO4 and other chemicals to control external parasites and other pathogens. However, accumulation of Cu in aquaculture ponds may pose a hazard to growth of fish or to the aquatic environment. The aim of this study was to determine accumulation of Cu in sediment, water and fish in a catfish pond with a history of repeated treatment with CuSO4 in the Mekong Delta, Vietnam. Copper concentrations in pond sediment were in the interval 21.3-45.7 mg kg(-1) dw and did not exceed the Vietnamese values for soil to be used for agricultural production (70 mg kg(-1) dw.). During three samplings the total mean concentration of Cu in pond water (4 μg L(-1)) did not exceed the LC50-value (70 μg L(-1)) for catfish and the mean dissolved concentration of Cu (0.986 μg L(-1)) did not seem to constitute a risk for the stability of the aquatic ecosystem. No significant variation in Cu concentrations between sampling sites in the pond and depth of sediment profile were determined. The accumulation of Cu in catfish was highest in the liver compared to the skin, gills and muscle tissue. With the current practice of removing pond sludge three to four times during a production cycle little if any Cu seems to accumulate in catfish ponds despite repeated anti-parasite treatments with CuSO4. Further studies are needed to assess the eco-toxicity and impact on agricultural production when pond sediment is discharged into aquatic recipients and used as soil fertilizer.

Microstructural evolution, mechanical properties, and strain hardening behavior of ultrafine grained commercial pure copper during the accumulative roll bonding process

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Fattah-alhosseini, A. [Department of Materials Engineering, Bu-Ali Sina University, Hamedan 65178-38695 (Iran, Islamic Republic of); Imantalab, O., E-mail: o.imantalab@gmail.com [Department of Materials Engineering, Bu-Ali Sina University, Hamedan 65178-38695 (Iran, Islamic Republic of); Mazaheri, Y. [Department of Materials Engineering, Bu-Ali Sina University, Hamedan 65178-38695 (Iran, Islamic Republic of); Keshavarz, M.K. [Department of Engineering Physics, Polytechnique Montreal, Montreal (Canada)

2016-01-05

In this study, the microstructural evolution, mechanical properties, and strain hardening behavior of commercial pure copper processed by the accumulative roll bonding (ARB) were investigated. Transmission electron microscopy (TEM) micrographs and atomic force microscopy (AFM) images indicated that with increasing the number of ARB cycles, the grain size of samples decreased. An Ultrafine grained (UFG) structure with an average grain size of about 200 nm was achieved after four cycles of ARB. The yield and ultimate tensile strength of pure copper with the UFG microstructure was reached about 360 MPa and 396 MPa (about 400% and 100% higher than that of the annealed state), respectively. All ARB-processed copper samples showed lower strain hardening exponent in comparison with the annealed state. Moreover, the strain hardening rate increased with increasing ARB cycles up to 3 cycles and then decreased.

Microstructural evolution, mechanical properties, and strain hardening behavior of ultrafine grained commercial pure copper during the accumulative roll bonding process

International Nuclear Information System (INIS)

Fattah-alhosseini, A.; Imantalab, O.; Mazaheri, Y.; Keshavarz, M.K.

2016-01-01

In this study, the microstructural evolution, mechanical properties, and strain hardening behavior of commercial pure copper processed by the accumulative roll bonding (ARB) were investigated. Transmission electron microscopy (TEM) micrographs and atomic force microscopy (AFM) images indicated that with increasing the number of ARB cycles, the grain size of samples decreased. An Ultrafine grained (UFG) structure with an average grain size of about 200 nm was achieved after four cycles of ARB. The yield and ultimate tensile strength of pure copper with the UFG microstructure was reached about 360 MPa and 396 MPa (about 400% and 100% higher than that of the annealed state), respectively. All ARB-processed copper samples showed lower strain hardening exponent in comparison with the annealed state. Moreover, the strain hardening rate increased with increasing ARB cycles up to 3 cycles and then decreased.

Evaluation of a commercially available molybdate formulation and zinc oxide boluses in preventing hepatic copper accumulation and thus enzootic icterus in sheep

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C.J. Botha

2001-07-01

Full Text Available The efficacy of a molybdate formulation and a zinc oxide bolus as prophylactic agents for enzootic icterus was evaluated in sheep. Before copper loading, liver biopsies were performed on 12 male, 6-month-old, Mutton Merino sheep to determine hepatic copper (Cu and zinc (Zn concentrations. The animals were restrictively randomised according to liver copper concentrations to 3 treatment groups (n = 4 to achieve similar mean liver copper concentrations per group. All sheep received 4 m /kg of a 0.5 %aqueous solution of CuSO4·5H2O intraruminally 7 days per week for 10 weeks. On Day 0 the sheep in the Mo-group were injected subcutaneously with 42 mg molybdenum (Mo contained in a commercial molybdate formulation. The animals in the Zn-group each received a zinc oxide bolus, containing 43 g zinc oxide, via a rumen cannula. Treatment was repeated on Day 42. Four animals served as untreated controls. Urinary copper excretion, plasma copper concentration, haematocrit and glutamate dehydrogenase (GLDH activity were determined throughout the trial. The animals were sacrificed after 10 weeks and liver samples were submitted for histopathological examination. Liver and kidney copper and zinc concentrations were determined. Neither the molybdate treatment nor the zinc oxide boluses prevented hepatic copper accumulation. The urinary copper excretion, plasma copper concentration, haematocrit and GLDH activity were not significantly different (P > 0.05 from the controls.

Dextran-Catechin: An anticancer chemically-modified natural compound targeting copper that attenuates neuroblastoma growth

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Vittorio, Orazio; Brandl, Miriam; Cirillo, Giuseppe; Kimpton, Kathleen; Hinde, Elizabeth; Gaus, Katharina; Yee, Eugene; Kumar, Naresh; Duong, Hien; Fleming, Claudia; Haber, Michelle; Norris, Murray; Boyer, Cyrille; Kavallaris, Maria

2016-01-01

Neuroblastoma is frequently diagnosed at advanced stage disease and treatment includes high dose chemotherapy and surgery. Despite the use of aggressive therapy survival rates are poor and children that survive their disease experience long term side effects from their treatment, highlighting the need for effective and less toxic therapies. Catechin is a natural polyphenol with anti-cancer properties and limited side effects, however its mechanism of action is unknown. Here we report that Dextran-Catechin, a conjugated form of catechin that increases serum stability, is preferentially and markedly active against neuroblastoma cells having high levels of intracellular copper, without affecting non-malignant cells. Copper transporter 1 (CTR1) is the main transporter of copper in mammalian cells and it is upregulated in neuroblastoma. Functional studies showed that depletion of CTR1 expression reduced intracellular copper levels and led to a decrease in neuroblastoma cell sensitivity to Dextran-Catechin, implicating copper in the activity of this compound. Mechanistically, Dextran-Catechin was found to react with copper, inducing oxidative stress and decreasing glutathione levels, an intracellular antioxidant and regulator of copper homeostasis. In vivo, Dextran-Catechin significantly attenuated tumour growth in human xenograft and syngeneic models of neuroblastoma. Thus, Dextran-Catechin targets copper, inhibits tumour growth, and may be valuable in the treatment of aggressive neuroblastoma and other cancers dependent on copper for their growth. PMID:27374085

Copper and Copper Proteins in Parkinson's Disease

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Rivera-Mancia, Susana; Diaz-Ruiz, Araceli; Tristan-Lopez, Luis; Rios, Camilo

2014-01-01

Copper is a transition metal that has been linked to pathological and beneficial effects in neurodegenerative diseases. In Parkinson's disease, free copper is related to increased oxidative stress, alpha-synuclein oligomerization, and Lewy body formation. Decreased copper along with increased iron has been found in substantia nigra and caudate nucleus of Parkinson's disease patients. Copper influences iron content in the brain through ferroxidase ceruloplasmin activity; therefore decreased protein-bound copper in brain may enhance iron accumulation and the associated oxidative stress. The function of other copper-binding proteins such as Cu/Zn-SOD and metallothioneins is also beneficial to prevent neurodegeneration. Copper may regulate neurotransmission since it is released after neuronal stimulus and the metal is able to modulate the function of NMDA and GABA A receptors. Some of the proteins involved in copper transport are the transporters CTR1, ATP7A, and ATP7B and the chaperone ATOX1. There is limited information about the role of those biomolecules in the pathophysiology of Parkinson's disease; for instance, it is known that CTR1 is decreased in substantia nigra pars compacta in Parkinson's disease and that a mutation in ATP7B could be associated with Parkinson's disease. Regarding copper-related therapies, copper supplementation can represent a plausible alternative, while copper chelation may even aggravate the pathology. PMID:24672633

Uptake and internalisation of copper by three marine microalgae: comparison of copper-sensitive and copper-tolerant species.

Science.gov (United States)

Levy, Jacqueline L; Angel, Brad M; Stauber, Jennifer L; Poon, Wing L; Simpson, Stuart L; Cheng, Shuk Han; Jolley, Dianne F

2008-08-29

Although it has been well established that different species of marine algae have different sensitivities to metals, our understanding of the physiological and biochemical basis for these differences is limited. This study investigated copper adsorption and internalisation in three algal species with differing sensitivities to copper. The diatom Phaeodactylum tricornutum was particularly sensitive to copper, with a 72-h IC50 (concentration of copper to inhibit growth rate by 50%) of 8.0 microg Cu L(-1), compared to the green algae Tetraselmis sp. (72-h IC50 47 microg Cu L(-1)) and Dunaliella tertiolecta (72-h IC50 530 microg Cu L(-1)). At these IC50 concentrations, Tetraselmis sp. had much higher intracellular copper (1.97+/-0.01 x 10(-13)g Cu cell(-1)) than P. tricornutum (0.23+/-0.19 x 10(-13)g Cu cell(-1)) and D. tertiolecta (0.59+/-0.05 x 10(-13)g Cu cell(-1)), suggesting that Tetraselmis sp. effectively detoxifies copper within the cell. By contrast, at the same external copper concentration (50 microg L(-1)), D. tertiolecta appears to better exclude copper than Tetraselmis sp. by having a slower copper internalisation rate and lower internal copper concentrations at equivalent extracellular concentrations. The results suggest that the use of internal copper concentrations and net uptake rates alone cannot explain differences in species-sensitivity for different algal species. Model prediction of copper toxicity to marine biota and understanding fundamental differences in species-sensitivity will require, not just an understanding of water quality parameters and copper-cell binding, but also further knowledge of cellular detoxification mechanisms.

Bacterial Killing by Dry Metallic Copper Surfaces▿

OpenAIRE

Santo, Christophe Espírito; Lam, Ee Wen; Elowsky, Christian G.; Quaranta, Davide; Domaille, Dylan W.; Chang, Christopher J.; Grass, Gregor

2010-01-01

Metallic copper surfaces rapidly and efficiently kill bacteria. Cells exposed to copper surfaces accumulated large amounts of copper ions, and this copper uptake was faster from dry copper than from moist copper. Cells suffered extensive membrane damage within minutes of exposure to dry copper. Further, cells removed from copper showed loss of cell integrity. Acute contact with metallic copper surfaces did not result in increased mutation rates or DNA lesions. These findings are important fir...

Copper-induced activation of TRP channels promotes extracellular calcium entry and activation of CaMs and CDPKs leading to copper entry and membrane depolarization in Ulva compressa

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Melissa eGómez

2015-03-01

Full Text Available In order to identify channels involved in membrane depolarization, Ulva compressa was incubated with agonists of TRP channels C5, A1 and V1 and the level of intracellular calcium was detected. Agonists of TRPC5, A1 and V1 induced increases in intracellular calcium at 4, 9 and 12 min of exposure, respectively, and antagonists of TRPC5, A1 and V1 corresponding to SKF-96365 (SKF, HC-030031 (HC and capsazepin (CPZ, respectively, inhibited calcium increases indicating that functional TRPs exist in U. compressa. In addition, copper excess induced increases in intracellular calcium at 4, 9 and 12 min which were inhibited by SKF, HC and CPZ, respectively, indicating that copper activate TRPC5, A1 and V1 channels. Moreover, copper-induced calcium increases were inhibited by EGTA, a non-permeable calcium chelating agent, but not by thapsigargin, an inhibitor of endoplasmic reticulum (ER calcium ATPase, indicating that activation of TRPs leads to extracellular calcium entry. Furthermore, copper-induced calcium increases were not inhibited by W-7, an inhibitor of CaMs, and staurosporine, an inhibitor of CDPKs, indicating that extracellular calcium entry did not require CaMs and CDPKs activation. In addition, copper induced membrane depolarization events at 4, 8 and 11 min and these events were inhibited by SKF, HC, CPZ and bathocuproine, a specific copper chelating agent, indicating copper entry through TRP channels leading to membrane depolarization. Moreover, membrane depolarization events were inhibited by W-7 and staurosporine, indicating that CaMs and CDPKs are required in order to activate TRPs to allow copper entry. Thus, light-dependent copper-induced activation TRPC5, A1 and V1 promotes extracellular calcium entry leading to activation of CaMs and CDPKs which, in turn, promotes copper entry through these TRP channels leading to membrane depolarization.

Copper economy in Chlamydomonas: Prioritized allocation and reallocation of copper to respiration vs. photosynthesis

Science.gov (United States)

Kropat, Janette; Gallaher, Sean D.; Urzica, Eugen I.; Nakamoto, Stacie S.; Strenkert, Daniela; Tottey, Stephen; Mason, Andrew Z.; Merchant, Sabeeha S.

2015-01-01

Inorganic elements, although required only in trace amounts, permit life and primary productivity because of their functions in catalysis. Every organism has a minimal requirement of each metal based on the intracellular abundance of proteins that use inorganic cofactors, but elemental sparing mechanisms can reduce this quota. A well-studied copper-sparing mechanism that operates in microalgae faced with copper deficiency is the replacement of the abundant copper protein plastocyanin with a heme-containing substitute, cytochrome (Cyt) c6. This switch, which is dependent on a copper-sensing transcription factor, copper response regulator 1 (CRR1), dramatically reduces the copper quota. We show here that in a situation of marginal copper availability, copper is preferentially allocated from plastocyanin, whose function is dispensable, to other more critical copper-dependent enzymes like Cyt oxidase and a ferroxidase. In the absence of an extracellular source, copper allocation to Cyt oxidase includes CRR1-dependent proteolysis of plastocyanin and quantitative recycling of the copper cofactor from plastocyanin to Cyt oxidase. Transcriptome profiling identifies a gene encoding a Zn-metalloprotease, as a candidate effecting copper recycling. One reason for the retention of genes encoding both plastocyanin and Cyt c6 in algal and cyanobacterial genomes might be because plastocyanin provides a competitive advantage in copper-depleted environments as a ready source of copper. PMID:25646490

Different roles of glutathione in copper and zinc chelation in Brassica napus roots.

Science.gov (United States)

Zlobin, Ilya E; Kartashov, Alexander V; Shpakovski, George V

2017-09-01

We investigated the specific features of copper and zinc excess action on the roots of canola (Brassica napus L.) plants. Copper rapidly accumulated in canola root cells and reached saturation during several hours of treatment, whereas the root zinc content increased relatively slowly. Excessive copper and zinc entry inside the cell resulted in significant cell damage, as evidenced by alterations in plasmalemma permeability and decreases in cellular enzymatic activity. Zinc excess specifically damaged root hair cells, which correlated with a pronounced elevation of their labile zinc level. In vitro, we showed that reduced glutathione (GSH) readily reacted with copper ions to form complexes with blocked sulfhydryl groups. In contrast, zinc ions were ineffective as glutathione blockers, and glutathione molecules did not lose their specific chemical activity in the presence of Zn 2+ ions. The effect of copper and zinc excess on the glutathione pool in canola root cells was analysed by a combination of biochemical determination of total and oxidized glutathione contents and fluorescent staining of free reduced glutathione with monochlorobimane dye. Excess copper led to dose-dependent diminution of free reduced glutathione contents in the root cells, which could not be explained by the loss of total cellular glutathione or its oxidation. In contrast, we observed little effect of much higher intracellular zinc concentrations on the free reduced glutathione content. We concluded that GSH plays an important role in copper excess, but not zinc excess chelation, in canola root cells. Copyright © 2017 Elsevier Masson SAS. All rights reserved.

Unexpected role of the copper transporter ATP7A in PDGF-induced vascular smooth

Energy Technology Data Exchange (ETDEWEB)

Ashino, T.; Varadarajan, S.; Urao, N.; Oshikawa, J.; Chen, G. -F.; Wang, H.; Huo, Y.; Finney, L.; Vogt, S.; McKinney, R. D.; Maryon, E. B.; Kaplan, J. H.; Ushio-Fukai, M.; Fukai, T. (Biosciences Division); ( XSD); ( PSC-USR); (Univ. of Illinois at Chicago); (Univ. of Minnesota)

2010-09-09

Copper, an essential nutrient, has been implicated in vascular remodeling and atherosclerosis with unknown mechanism. Bioavailability of intracellular copper is regulated not only by the copper importer CTR1 (copper transporter 1) but also by the copper exporter ATP7A (Menkes ATPase), whose function is achieved through copper-dependent translocation from trans-Golgi network (TGN). Platelet-derived growth factor (PDGF) promotes vascular smooth muscle cell (VSMC) migration, a key component of neointimal formation. To determine the role of copper transporter ATP7A in PDGF-induced VSMC migration. Depletion of ATP7A inhibited VSMC migration in response to PDGF or wound scratch in a CTR1/copper-dependent manner. PDGF stimulation promoted ATP7A translocation from the TGN to lipid rafts, which localized at the leading edge, where it colocalized with PDGF receptor and Rac1, in migrating VSMCs. Mechanistically, ATP7A small interfering RNA or CTR small interfering RNA prevented PDGF-induced Rac1 translocation to the leading edge, thereby inhibiting lamellipodia formation. In addition, ATP7A depletion prevented a PDGF-induced decrease in copper level and secretory copper enzyme precursor prolysyl oxidase (Pro-LOX) in lipid raft fraction, as well as PDGF-induced increase in LOX activity. In vivo, ATP7A expression was markedly increased and copper accumulation was observed by synchrotron-based x-ray fluorescence microscopy at neointimal VSMCs in wire injury model. These findings suggest that ATP7A plays an important role in copper-dependent PDGF-stimulated VSMC migration via recruiting Rac1 to lipid rafts at the leading edge, as well as regulating LOX activity. This may contribute to neointimal formation after vascular injury. Our findings provide insight into ATP7A as a novel therapeutic target for vascular remodeling and atherosclerosis.

Accumulation of lead, zinc, copper and cadmium by 12 wetland plant species thriving in metal-contaminated sites in China

International Nuclear Information System (INIS)

Deng, H.; Ye, Z.H.; Wong, M.H.

2004-01-01

The concentrations of lead, zinc, copper and cadmium accumulated by 12 emergent-rooted wetland plant species including different populations of Leersia hexandra, Juncus effusus and Equisetum ramosisti were investigated in field conditions of China. The results showed that metal accumulation by wetland plants differed among species, populations and tissues. Populations grown in substrata with elevated metals contained significantly higher metals in plants. Metals accumulated by wetland plants were mostly distributed in root tissues, suggesting that an exclusion strategy for metal tolerance widely exists in them. That some species/populations could accumulate relatively high metal concentrations (far above the toxic concentration to plants) in their shoots indicates that internal detoxification metal tolerance mechanism(s) are also included. The factors affecting metal accumulation by wetland plants include metal concentrations, pH, and nutrient status in substrata. Mostly concentrations of Pb and Cu in both aboveground and underground tissues of the plants were significantly positively related to their total and/or DTPA-extractable fractions in substrata while negatively to soil N and P, respectively. The potential use of these wetland plants in phytoremediation is also discussed

Accumulation of lead, zinc, copper and cadmium by 12 wetland plant species thriving in metal-contaminated sites in China

Energy Technology Data Exchange (ETDEWEB)

Deng, H.; Ye, Z.H.; Wong, M.H

2004-11-01

The concentrations of lead, zinc, copper and cadmium accumulated by 12 emergent-rooted wetland plant species including different populations of Leersia hexandra, Juncus effusus and Equisetum ramosisti were investigated in field conditions of China. The results showed that metal accumulation by wetland plants differed among species, populations and tissues. Populations grown in substrata with elevated metals contained significantly higher metals in plants. Metals accumulated by wetland plants were mostly distributed in root tissues, suggesting that an exclusion strategy for metal tolerance widely exists in them. That some species/populations could accumulate relatively high metal concentrations (far above the toxic concentration to plants) in their shoots indicates that internal detoxification metal tolerance mechanism(s) are also included. The factors affecting metal accumulation by wetland plants include metal concentrations, pH, and nutrient status in substrata. Mostly concentrations of Pb and Cu in both aboveground and underground tissues of the plants were significantly positively related to their total and/or DTPA-extractable fractions in substrata while negatively to soil N and P, respectively. The potential use of these wetland plants in phytoremediation is also discussed.

Triiodothyronine enhances accumulation of intracellular lipids in adipocytes through thyroid hormone receptor α via direct and indirect mechanisms.

Science.gov (United States)

Gambo, Yurina; Matsumura, Miki; Fujimori, Ko

2016-08-15

Triiodothyronine (T3) enhanced the expression of adipogenic and lipogenic genes with elevation of the intracellular lipids through thyroid hormone receptor (TR) α in mouse 3T3-L1 cells. However, the transcription of the SREBP-1c and HSL genes was decreased by T3. Such T3-mediated alterations were negated by TRα siRNA. Chromatin immunoprecipitation assay showed that the binding of TRα to the TR-responsive element (TRE) of the FAS promoter was elevated by T3. In contrast, the ability of TRα to bind to the TRE of the SREBP-1c promoter was decreased by T3. In addition, the binding of SREBP-1c to the SRE of the HSL promoter was lowered by T3. These results indicate that T3 increased the accumulation of intracellular lipids by enhancing the expression of the FAS gene through direct binding of TRα to the FAS promoter and simultaneously lowered the amount of lipolysis via reduced binding of T3-decreased SREBP-1c to the HSL promoter. Copyright © 2016 Elsevier Ireland Ltd. All rights reserved.

Copper and Copper Proteins in Parkinson’s Disease

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Sergio Montes

2014-01-01

Full Text Available Copper is a transition metal that has been linked to pathological and beneficial effects in neurodegenerative diseases. In Parkinson’s disease, free copper is related to increased oxidative stress, alpha-synuclein oligomerization, and Lewy body formation. Decreased copper along with increased iron has been found in substantia nigra and caudate nucleus of Parkinson’s disease patients. Copper influences iron content in the brain through ferroxidase ceruloplasmin activity; therefore decreased protein-bound copper in brain may enhance iron accumulation and the associated oxidative stress. The function of other copper-binding proteins such as Cu/Zn-SOD and metallothioneins is also beneficial to prevent neurodegeneration. Copper may regulate neurotransmission since it is released after neuronal stimulus and the metal is able to modulate the function of NMDA and GABA A receptors. Some of the proteins involved in copper transport are the transporters CTR1, ATP7A, and ATP7B and the chaperone ATOX1. There is limited information about the role of those biomolecules in the pathophysiology of Parkinson’s disease; for instance, it is known that CTR1 is decreased in substantia nigra pars compacta in Parkinson’s disease and that a mutation in ATP7B could be associated with Parkinson’s disease. Regarding copper-related therapies, copper supplementation can represent a plausible alternative, while copper chelation may even aggravate the pathology.

Unraveling the Amycolatopsis tucumanensis copper-resistome.

Science.gov (United States)

Dávila Costa, José Sebastián; Kothe, Erika; Abate, Carlos Mauricio; Amoroso, María Julia

2012-10-01

Heavy metal pollution is widespread causing serious ecological problems in many parts of the world; especially in developing countries where a budget for remediation technology is not affordable. Therefore, screening for microbes with high accumulation capacities and studying their stable resistance characteristics is advisable to define cost-effective any remediation strategies. Herein, the copper-resistome of the novel copper-resistant strain Amycolatopsis tucumanensis was studied using several approaches. Two dimensional gel electrophoresis revealed that proteins of the central metabolism, energy production, transcriptional regulators, two-component system, antioxidants and protective metabolites increased their abundance upon copper-stress conditions. Transcriptome analysis revealed that in presence of copper, superoxide dismutase, alkyl hydroperoxide reductase and mycothiol reductase genes were markedly induced in expression. The oxidative damage of protein and lipid from A. tucumanensis was negligible compared with that observed in the copper-sensitive strain Amycolatopsis eurytherma. Thus, we provide evidence that A. tucumamensis shows a high adaptation towards copper, the sum of which is proposed as the copper-resistome. This adaptation allows the strain to accumulate copper and survive this stress; besides, it constitutes the first report in which the copper-resistome of a strain of the genus Amycolatopsis with bioremediation potential has been evaluated.

Remediation of copper in vineyards – A mini review

International Nuclear Information System (INIS)

Mackie, K.A.; Müller, T.; Kandeler, E.

2012-01-01

Viticulturists use copper fungicide to combat Downy Mildew. Copper, a non-degradable heavy metal, can accumulate in soil or leach into water sources. Its accumulation in topsoil has impacted micro and macro organisms, spurring scientists to research in situ copper removal methods. Recent publications suggest that microorganism assisted phytoextraction, using plants and bacteria to actively extract copper, is most promising. As vineyards represent moderately polluted sites this technique has great potential. Active plant extraction and chelate assisted remediation extract too little copper or risk leaching, respectively. However, despite interesting pot experiment results using microorganism assisted phytoextraction, it remains a challenge to find plants that primarily accumulate copper in their shoots, a necessity in vineyards where whole plant removal would be time consuming and financially cumbersome. Vineyard remediation requires a holistic approach including sustainable soil management, proper plant selection, increasing biodiversity and microorganisms. - Highlights: ► We describe copper distribution and availability in vineyards. ► We explain the environmental impact of copper on organisms, plants and processes. ► We detail possible remediation methods within vineyards. ► Microbially assisted phytoremediation is the most promising remediation method. ► A solution requires an interdisciplinary approach between plants, soil and vines. - This review is significant because it highlights prospective remediation methods usable in copper contaminated vineyards.

Copper metabolism: a multicompartmental model of copper kinetics in the rat

International Nuclear Information System (INIS)

Dunn, M.A.

1985-01-01

A qualitative multicompartmental model was developed that describes the whole-body kinetics of copper metabolism in the adult rat. The model was developed from radiocopper percent dose vs. time data measured over a three day period in plasma, liver, skin, skeletal muscle, bile and feces after the intravenous injection of 10 μg copper labeled with 64 Cu. Plasma radiocopper was separated into ceruloplasmin (Cp) and nonceruloplasmin (NCp) fractions. Liver cytosolic radiocopper was fractionated into void volume superoxide dismutase (SOD) containing and metallothionein fractions by gel filtration. Liver particulate fractions were isolated by differential centrifugation. The SAAM and CONSAM modeling programs were used to develop the model. The sizes of compartments, fractional rate constants and mass transfer rates between compartments were evaluated. The intracellular metabolism of copper was similar in hepatic and extrahepatic tissues being comprised of a faster turning over compartment (FTC) exchanging copper with NCp and a slower turning over compartment (STC) with input from Cp. Output from the STC was into the FTC. In the liver the STC was postulated to represent SOD copper which unlike the extrahepatic tissues received much of its input from the FTC. A small amount of biliary copper (9%) was postulated to return to plasma NCp by enterohepatic recycling. The model developed was contrasted and compared with two previous models of copper metabolism

Role of ATP-binding cassette and solute carrier transporters in erlotinib CNS penetration and intracellular accumulation.

Science.gov (United States)

Elmeliegy, Mohamed A; Carcaboso, Angel M; Tagen, Michael; Bai, Feng; Stewart, Clinton F

2011-01-01

To study the role of drug transporters in central nervous system (CNS) penetration and cellular accumulation of erlotinib and its metabolite, OSI-420. After oral erlotinib administration to wild-type and ATP-binding cassette (ABC) transporter-knockout mice (Mdr1a/b(-/-), Abcg2(-/-), Mdr1a/b(-/-)Abcg2(-/-), and Abcc4(-/-)), plasma was collected and brain extracellular fluid (ECF) was sampled using intracerebral microdialysis. A pharmacokinetic model was fit to erlotinib and OSI-420 concentration-time data, and brain penetration (P(Brain)) was estimated by the ratio of ECF-to-unbound plasma area under concentration-time curves. Intracellular accumulation of erlotinib was assessed in cells overexpressing human ABC transporters or SLC22A solute carriers. P(Brain) in wild-type mice was 0.27 ± 0.11 and 0.07 ± 0.02 (mean ± SD) for erlotinib and OSI-420, respectively. Erlotinib and OSI-420 P(Brain) in Abcg2(-/-) and Mdr1a/b(-/-)Abcg2(-/-) mice were significantly higher than in wild-type mice. Mdr1a/b(-/-) mice showed similar brain ECF penetration as wild-type mice (0.49 ± 0.37 and 0.04 ± 0.02 for erlotinib and OSI-420, respectively). In vitro, erlotinib and OSI-420 accumulation was significantly lower in cells overexpressing breast cancer resistance protein (BCRP) than in control cells. Only OSI-420, not erlotinib, showed lower accumulation in cells overexpressing P-glycoprotein (P-gp) than in control cells. The P-gp/BCRP inhibitor elacridar increased erlotinib and OSI-420 accumulation in BCRP-overexpressing cells. Erlotinib uptake was higher in OAT3- and OCT2-transfected cells than in empty vector control cells. Abcg2 is the main efflux transporter preventing erlotinib and OSI-420 penetration in mouse brain. Erlotinib and OSI-420 are substrates for SLC22A family members OAT3 and OCT2. Our findings provide a mechanistic basis for erlotinib CNS penetration, cellular uptake, and efflux mechanisms. ©2010 AACR.

Copper: From neurotransmission to neuroproteostasis

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Carlos M Opazo

2014-07-01

Full Text Available Copper is critical for the Central Nervous System (CNS development and function. In particular, different studies have shown the effect of copper at brain synapses, where it inhibits Long Term Potentation (LTP and receptor pharmacology. Paradoxically, according to recent studies copper is required for a normal LTP response. Copper is released at the synaptic cleft, where it blocks glutamate receptors, which explain its blocking effects on excitatory neurotransmission. Our results indicate that copper also enhances neurotransmission through the accumulation of PSD95 protein, which increase the levels of AMPA receptors located at the plasma membrane of the post-synaptic density. Thus, our findings represent a novel mechanism for the action of copper, which may have implications for the neurophysiology and neuropathology of the CNS. These data indicate that synaptic configuration is sensitive to transient changes in transition metal homeostasis. Our results suggest that copper increases GluA1 subunit levels of the AMPA receptor through the anchorage of AMPA receptors to the plasma membrane as a result of PSD-95 accumulation. Here, we will review the role of copper on neurotransmission of CNS neurons. In addition, we will discuss the potential mechanisms by which copper could modulate neuronal proteostasis (neuroproteostasis in the CNS with focus in the Ubiquitin Proteasome System, which is particularly relevant to neurological disorders such Alzheimer’s disease (AD where copper and protein dyshomeostasis may contribute to neurodegeneration. An understanding of these mechanisms may ultimately lead to the development of novel therapeutic approaches to control metal and synaptic alterations observed in AD patients.

Copper-induced apical trafficking of ATP7B in polarized hepatoma cells provides a mechanism for biliary copper excretion

NARCIS (Netherlands)

Roelofsen, H; Wolters, H; Van Luyn, MJA; Miura, N; Kuipers, F; Vonk, RJ

Background & Aims: Mutations in the ATP7B gene, encoding a copper-transporting P-type adenosine triphosphatase, lead to excessive hepatic copper accumulation because of impaired biliary copper excretion in Wilson's disease. In human liver, ATP7B is predominantly localized to the trans-Golgi network,

Copper isotope fractionation by desert shrubs

International Nuclear Information System (INIS)

Navarrete, Jesica U.; Viveros, Marian; Ellzey, Joanne T.; Borrok, David M.

2011-01-01

Copper has two naturally occurring stable isotopes of masses 63 and 65 which can undergo mass dependent fractionation during various biotic and abiotic chemical reactions. These interactions and their resulting Cu isotope fractionations can be used to determine the mechanisms involved in the cycling of Cu in natural systems. In this study, Cu isotope changes were investigated at the organismal level in the metal-accumulating desert plant, Prosopis pubescens. Initial results suggest that the lighter Cu isotope was preferentially incorporated into the leaves of the plant, which may suggest that Cu was actively transported via intracellular proteins. The roots and stems show a smaller degree of Cu isotope fractionation and the direction and magnitude of the fractionations was dependent upon the levels of Cu exposure. Based on this and previous work with bacteria and yeast, a trend is emerging that suggests the lighter Cu isotope is preferentially incorporated into biological components, while the heavier Cu isotope tends to become enriched in aqueous solutions. In bacteria, plants and animals, intracellular Cu concentrations are strictly regulated via dozens of enzymes that can bind, transport, and store Cu. Many of these enzymes reduce Cu(II) to Cu(I). These initial results seem to fit into a broader picture of Cu isotope cycling in natural systems where oxidation/reduction reactions are fundamental in controlling the distributions of Cu isotopes.

Aspergillus fumigatus Copper Export Machinery and Reactive Oxygen Intermediate Defense Counter Host Copper-Mediated Oxidative Antimicrobial Offense

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Philipp Wiemann

2017-05-01

Full Text Available The Fenton-chemistry-generating properties of copper ions are considered a potent phagolysosome defense against pathogenic microbes, yet our understanding of underlying host/microbe dynamics remains unclear. We address this issue in invasive aspergillosis and demonstrate that host and fungal responses inextricably connect copper and reactive oxygen intermediate (ROI mechanisms. Loss of the copper-binding transcription factor AceA yields an Aspergillus fumigatus strain displaying increased sensitivity to copper and ROI in vitro, increased intracellular copper concentrations, decreased survival in challenge with murine alveolar macrophages (AMΦs, and reduced virulence in a non-neutropenic murine model. ΔaceA survival is remediated by dampening of host ROI (chemically or genetically or enhancement of copper-exporting activity (CrpA in A. fumigatus. Our study exposes a complex host/microbe multifactorial interplay that highlights the importance of host immune status and reveals key targetable A. fumigatus counter-defenses.

Copper Tolerance and Biosorption of Saccharomyces cerevisiae during Alcoholic Fermentation

Science.gov (United States)

Liu, Ling-ling; Jia, Bo; Zhao, Fang; Huang, Wei-dong; Zhan, Ji-cheng

2015-01-01

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. PMID:26030864

Copper Tolerance and Biosorption of Saccharomyces cerevisiae during Alcoholic Fermentation.

Directory of Open Access Journals (Sweden)

Xiang-Yu Sun

Full Text Available 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.

Inhibition of P-glycoprotein by HIV protease inhibitors increases intracellular accumulation of berberine in murine and human macrophages.

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Weibin Zha

Full Text Available HIV protease inhibitor (PI-induced inflammatory response in macrophages is a major risk factor for cardiovascular diseases. We have previously reported that berberine (BBR, a traditional herbal medicine, prevents HIV PI-induced inflammatory response through inhibiting endoplasmic reticulum (ER stress in macrophages. We also found that HIV PIs significantly increased the intracellular concentrations of BBR in macrophages. However, the underlying mechanisms of HIV PI-induced BBR accumulation are unknown. This study examined the role of P-glycoprotein (P-gp in HIV PI-mediated accumulation of BBR in macrophages.Cultured mouse RAW264.7 macrophages, human THP-1-derived macrophages, Wild type MDCK (MDCK/WT and human P-gp transfected (MDCK/P-gp cells were used in this study. The intracellular concentration of BBR was determined by HPLC. The activity of P-gp was assessed by measuring digoxin and rhodamine 123 (Rh123 efflux. The interaction between P-gp and BBR or HIV PIs was predicated by Glide docking using Schrodinger program. The results indicate that P-gp contributed to the efflux of BBR in macrophages. HIV PIs significantly increased BBR concentrations in macrophages; however, BBR did not alter cellular HIV PI concentrations. Although HIV PIs did not affect P-gp expression, P-gp transport activities were significantly inhibited in HIV PI-treated macrophages. Furthermore, the molecular docking study suggests that both HIV PIs and BBR fit the binding pocket of P-gp, and HIV PIs may compete with BBR to bind P-gp.HIV PIs increase the concentration of BBR by modulating the transport activity of P-gp in macrophages. Understanding the cellular mechanisms of potential drug-drug interactions is critical prior to applying successful combinational therapy in the clinic.

Leach-SX-EW copper revalorization from overburden of abandoned copper mine Cerovo, Eastern Serbia

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Stevanović Z.

2009-01-01

Full Text Available Hydrometallurgical processes for copper revalorization from overburden of abandoned mine Cerovo in Eastern Serbia were studied. Paper contain results of percolation leaching tests, performed with acidic mine waters accumulated in the bottom of the former open pit, followed by solvent extraction (SX and electrowinning (EW processes on achieved copper pregnant leach solutions. Usage of accumulated waste waters was objected to minimizing the environmental hazard due to uncontrolled leaking of these waters in nearby creeks and rivers. Chemical composition of acidic mine waters used for leaching tests was: (g/dm3: Cu - 0.201; Fe - 0.095; Mn - 0.041; Zn - 0.026; Ni - 0.0004; pH value - 3.3. Copper content in overburden sample used for leaching tests was 0.21% from which 64% were oxide copper minerals. In scope of leaching tests were examined influence of leaching solution pH values and iron (III concentration on copper recovery. It was established that for 120 hours of leaching on pH=1.5 without oxidant agents, copper concentration in pregnant leach solutions enriched up to 1.08g/dm3 which was enough for copper extraction from solution with SX-EW treatment. As extraction reagent in SX circuit was used LIX-984N in a kerosene diluent. Cathode current density in electrowinning cell was 220Am-2 while electrolyte temperature was kept on 50±2oC. Produced cathode copper at the end of SX-EW process has purity of 99.95% Cu.

Exogenous addition of histidine reduces copper availability in the yeast Saccharomyces cerevisiae

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Daisuke Watanabe

2014-07-01

Full Text Available The basic amino acid histidine inhibited yeast cell growth more severely than lysine and arginine. Overexpression of CTR1, which encodes a high-affinity copper transporter on the plasma membrane, or addition of copper to the medium alleviated this cytotoxicity. However, the intracellular level of copper ions was not decreased in the presence of excess histidine. These results indicate that histidine cytotoxicity is associated with low copper availability inside cells, not with impaired copper uptake. Furthermore, histidine did not affect cell growth under limited respiration conditions, suggesting that histidine cytotoxicity is involved in deficiency of mitochondrial copper.

Exogenous addition of histidine reduces copper availability in the yeast Saccharomyces cerevisiae.

Science.gov (United States)

Watanabe, Daisuke; Kikushima, Rie; Aitoku, Miho; Nishimura, Akira; Ohtsu, Iwao; Nasuno, Ryo; Takagi, Hiroshi

2014-07-07

The basic amino acid histidine inhibited yeast cell growth more severely than lysine and arginine. Overexpression of CTR1 , which encodes a high-affinity copper transporter on the plasma membrane, or addition of copper to the medium alleviated this cytotoxicity. However, the intracellular level of copper ions was not decreased in the presence of excess histidine. These results indicate that histidine cytotoxicity is associated with low copper availability inside cells, not with impaired copper uptake. Furthermore, histidine did not affect cell growth under limited respiration conditions, suggesting that histidine cytotoxicity is involved in deficiency of mitochondrial copper.

A Thapsigargin-Resistant Intracellular Calcium Sequestering Compartment in Rat Brain

Science.gov (United States)

2000-03-31

have a major impact on neuronal intracellular signaling. Most of the ER in neurons and glia appears to accumulate calcium by energy driven ion pumps...secretion of exocrine, endocrine, and neurocrine products, regulation of glycogenolysis and gluconeogenesis , intracellular transport, secretion of fluids...the RyRs [140]. Furthermore, the intracellular expression of these receptor-channels in neuronal ER is also reciprocal with RyRs located primarily in

Metavanadate causes cellular accumulation of copper and decreased lysyl oxidase activity

International Nuclear Information System (INIS)

Cui, Changtai T.; Uriu-Adams, Janet Y.; Tchaparian, Eskouhie H.; Keen, Carl L.; Rucker, Robert B.

2004-01-01

Selected indices of copper metabolism in weanling rats and fibroblast cultures were progressively altered in response to increased levels of sodium metavanadate. In diets, vanadium was added in amounts ranging from 0 to 80 μg V/g of diet, that is, 0-1.6 μmol V/g of diet. In fibroblast cultures, vanadium ranged from 0 to 400 nmol V/ml. The inhibition of P-ATPase-7A activity by metavanadate, important to copper egress from cells, was a primary focus. In skin, and tendon, the copper concentration was increased in response to increased dietary levels of metavanadate, whereas lysyl oxidase activity, a secreted cuproprotein, was reduced. The reduction in lysyl oxidase activity was also accompanied by reduced redox cycling potential of isolated fractions of lysyl oxidase, presumably due to reduced lysyltyrosyl quinone (LTQ) formation at the active site of lysyl oxidase. In contrast, liver copper concentrations and plasma ceruloplasmin activity were not affected by metavanadate exposure. However, semicarbazide-sensitive benzylamine oxidase (SCBO) activity, which was taken as an indirect measure of vascular adhesive protein-1 (VAP-1), was increased. In cultured fibroblasts, cellular copper was also increased and lysyl oxidase decreased in response to metavanadate. Moreover, the steady-state levels of atp7a and lysyl oxidase mRNAs were not affected by addition of metavanadate to culture medium up to 200 nmol/ml. Taken together, these data suggest that pathways involving copper egress and lysyl oxidase activation are particularly sensitive to metavanadate exposure through processes that are predominately posttranslational

Copper-associated hepatitis in dogs; pathogenesis, diagnosis and treatment

NARCIS (Netherlands)

Dirksen, K.|info:eu-repo/dai/nl/412424428

2016-01-01

Copper is an essential trace element for living organisms, but can have deleterious consequences when present in excess. Because the liver has a central role in copper metabolism, this is the predominant organ affected. Copper-accumulating disorders are recognized as hereditary diseases in man and

Intracellular mechanisms of solar water disinfection

Science.gov (United States)

Castro-Alférez, María; Polo-López, María Inmaculada; Fernández-Ibáñez, Pilar

2016-12-01

Solar water disinfection (SODIS) is a zero-cost intervention measure to disinfect drinking water in areas of poor access to improved water sources, used by more than 6 million people in the world. The bactericidal action of solar radiation in water has been widely proven, nevertheless the causes for this remain still unclear. Scientific literature points out that generation of reactive oxygen species (ROS) inside microorganisms promoted by solar light absorption is the main reason. For the first time, this work reports on the experimental measurement of accumulated intracellular ROS in E. coli during solar irradiation. For this experimental achievement, a modified protocol based on the fluorescent probe dichlorodihydrofluorescein diacetate (DCFH-DA), widely used for oxidative stress in eukaryotic cells, has been tested and validated for E. coli. Our results demonstrate that ROS and their accumulated oxidative damages at intracellular level are key in solar water disinfection.

The effects of copper proximity on oxalate production in Fibroporia radiculosa

Science.gov (United States)

Katie M. Jenkins; Carol A. Clausen; Frederick Green III

2014-01-01

Copper remains a key component used in wood preservatives available today. However, the observed tolerance of several critical wood rotting organisms continues to be problematic. Tolerance to copper has been linked to the production and accumulation of oxalate, which precipitates copper into insoluble copper-oxalate crystals, thus inactivating copper ions. The purpose...

Comparative genomic analyses of copper transporters and cuproproteomes reveal evolutionary dynamics of copper utilization and its link to oxygen.

Directory of Open Access Journals (Sweden)

Perry G Ridge

2008-01-01

Full Text Available Copper is an essential trace element in many organisms and is utilized in all domains of life. It is often used as a cofactor of redox proteins, but is also a toxic metal ion. Intracellular copper must be carefully handled to prevent the formation of reactive oxygen species which pose a threat to DNA, lipids, and proteins. In this work, we examined patterns of copper utilization in prokaryotes by analyzing the occurrence of copper transporters and copper-containing proteins. Many organisms, including those that lack copper-dependent proteins, had copper exporters, likely to protect against copper ions that inadvertently enter the cell. We found that copper use is widespread among prokaryotes, but also identified several phyla that lack cuproproteins. This is in contrast to the use of other trace elements, such as selenium, which shows more scattered and reduced usage, yet larger selenoproteomes. Copper transporters had different patterns of occurrence than cuproproteins, suggesting that the pathways of copper utilization and copper detoxification are independent of each other. We present evidence that organisms living in oxygen-rich environments utilize copper, whereas the majority of anaerobic organisms do not. In addition, among copper users, cuproproteomes of aerobic organisms were larger than those of anaerobic organisms. Prokaryotic cuproproteomes were small and dominated by a single protein, cytochrome c oxidase. The data are consistent with the idea that proteins evolved to utilize copper following the oxygenation of the Earth.

Effects of copper on the sabellid polychaete, Eudistylia vancouveri. II. copper accumulation and tissue injury in the branchial crown

Energy Technology Data Exchange (ETDEWEB)

Young, J S [Pacific Northwest Lab., Sequim, WA; Adee, R R; Piscopo, I; Buschbom, R L

1981-01-01

Copper in seawater caused injury to the radioles (gills) of the sabellid polychaete, Eudistylia vancouveri. Light and electron microscopy showed the loss of cellular adhesion and the structural derangement that lead to cell necrosis and death. The progression of injury was related to the uptake of copper into the tissues. Copper was found by X-ray microanalysis to be localized subcellularly in membrane-bound vesicles that are similar to lysosomes. Cell breakdown may result from lysosmal labilization.

Regulation of the copper chaperone CCS by XIAP-mediated ubiquitination.

Science.gov (United States)

Brady, Graham F; Galbán, Stefanie; Liu, Xuwen; Basrur, Venkatesha; Gitlin, Jonathan D; Elenitoba-Johnson, Kojo S J; Wilson, Thomas E; Duckett, Colin S

2010-04-01

In order to balance the cellular requirements for copper with its toxic properties, an elegant set of mechanisms has evolved to regulate and buffer intracellular copper. The X-linked inhibitor of apoptosis (XIAP) protein was recently identified as a copper-binding protein and regulator of copper homeostasis, although the mechanism by which XIAP binds copper in the cytosol is unclear. Here we describe the identification of the copper chaperone for superoxide dismutase (CCS) as a mediator of copper delivery to XIAP in cells. We also find that CCS is a target of the E3 ubiquitin ligase activity of XIAP, although interestingly, ubiquitination of CCS by XIAP was found to lead to enhancement of its chaperone activity toward its physiologic target, superoxide dismutase 1, rather than proteasomal degradation. Collectively, our results reveal novel links among apoptosis, copper metabolism, and redox regulation through the XIAP-CCS complex.

Intracellular Ca2+ Regulation in Calcium Sensitive Phenotype of Saccharomyces cerevisiae

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HERMANSYAH

2010-03-01

Full Text Available Intracellular cytosolic Ca2+ concentration accumulation plays an essential information in Saccharomyces cerevisiae i.e. to explain cellular mechanism of Ca2+ sensitive phenotype. Disruption both S. cerevisiae PPase PTP2 and MSG5 genes showed an inhibited growth in the presence of Ca2+. On the other hand, by using Luminocounter with apoaequorin system, a method based upon luminescent photoprotein aequorin, intracellular Ca2+ concentration was accumulated as a consequence of calcium sensitive phenotype of S. cerevisiae. This fact indicated that PPase ptp2Δ and msg5Δ were involved in intracellular Ca2+ transport in addition their already known pathways i.e Mitogen Activated Protein Kinase cell wall integrity pathway, high osmolarity glycerol (HOG pathway, and pheromone response FUS3 pathway.

Differences in Copper Absorption and Accumulation between Copper-Exclusion and Copper-Enrichment Plants: A Comparison of Structure and Physiological Responses.

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Fu, Lei; Chen, Chen; Wang, Bin; Zhou, Xishi; Li, Shuhuan; Guo, Pan; Shen, Zhenguo; Wang, Guiping; Chen, Yahua

2015-01-01

Differences in copper (Cu) absorption and transport, physiological responses and structural characteristics between two types of Cu-resistant plants, Oenothera glazioviana (Cu-exclusion type) and Elsholtzia haichowensis (Cu-enrichment type), were investigated in the present study. The results indicated the following: (1) After 50 μM Cu treatment, the Cu ratio in the xylem vessels of E. haichowensis increased by 60%. A Cu adsorption experiment indicated that O. glazioviana exhibited greater resistance to Cu, and Cu absorption and the shoot/root ratio of Cu were significantly lower in O. glazioviana than in E. haichowensis. (2) An analysis of the endogenous abscisic acid (ABA) variance and exogenous ABA treatment demonstrated that the ABA levels of both plants did not differ; exogenous ABA treatment clearly reduced Cu accumulation in both plants. (3) The leaf stomatal density of O. glazioviana was significantly less than that of E. haichowensis. Guard cells in E. haichowensis plants were covered with a thick cuticle layer, the epidermal hair was more numerous and longer, and the number of xylem conduits in the root was small. (4) The transpiration rate and the stomatal conductance of O. glazioviana were both significantly lower than those of E. haichowensis, regardless of whether the plants were treated with Cu. Taken together, these results indicate that the differences in the structural characteristics between these two plant species, particularly in the characteristics related to plant transpiration, are important factors that govern whether plants acquire or exclude Cu.

Differences in Copper Absorption and Accumulation between Copper-Exclusion and Copper-Enrichment Plants: A Comparison of Structure and Physiological Responses

Science.gov (United States)

Fu, Lei; Chen, Chen; Wang, Bin; Zhou, Xishi; Li, Shuhuan; Guo, Pan; Shen, Zhenguo; Wang, Guiping; Chen, Yahua

2015-01-01

Differences in copper (Cu) absorption and transport, physiological responses and structural characteristics between two types of Cu-resistant plants, Oenothera glazioviana (Cu-exclusion type) and Elsholtzia haichowensis (Cu-enrichment type), were investigated in the present study. The results indicated the following: (1) After 50 μM Cu treatment, the Cu ratio in the xylem vessels of E. haichowensis increased by 60%. A Cu adsorption experiment indicated that O. glazioviana exhibited greater resistance to Cu, and Cu absorption and the shoot/root ratio of Cu were significantly lower in O. glazioviana than in E. haichowensis. (2) An analysis of the endogenous abscisic acid (ABA) variance and exogenous ABA treatment demonstrated that the ABA levels of both plants did not differ; exogenous ABA treatment clearly reduced Cu accumulation in both plants. (3) The leaf stomatal density of O. glazioviana was significantly less than that of E. haichowensis. Guard cells in E. haichowensis plants were covered with a thick cuticle layer, the epidermal hair was more numerous and longer, and the number of xylem conduits in the root was small. (4) The transpiration rate and the stomatal conductance of O. glazioviana were both significantly lower than those of E. haichowensis, regardless of whether the plants were treated with Cu. Taken together, these results indicate that the differences in the structural characteristics between these two plant species, particularly in the characteristics related to plant transpiration, are important factors that govern whether plants acquire or exclude Cu. PMID:26207743

Differences in Copper Absorption and Accumulation between Copper-Exclusion and Copper-Enrichment Plants: A Comparison of Structure and Physiological Responses.

Directory of Open Access Journals (Sweden)

Lei Fu

Full Text Available Differences in copper (Cu absorption and transport, physiological responses and structural characteristics between two types of Cu-resistant plants, Oenothera glazioviana (Cu-exclusion type and Elsholtzia haichowensis (Cu-enrichment type, were investigated in the present study. The results indicated the following: (1 After 50 μM Cu treatment, the Cu ratio in the xylem vessels of E. haichowensis increased by 60%. A Cu adsorption experiment indicated that O. glazioviana exhibited greater resistance to Cu, and Cu absorption and the shoot/root ratio of Cu were significantly lower in O. glazioviana than in E. haichowensis. (2 An analysis of the endogenous abscisic acid (ABA variance and exogenous ABA treatment demonstrated that the ABA levels of both plants did not differ; exogenous ABA treatment clearly reduced Cu accumulation in both plants. (3 The leaf stomatal density of O. glazioviana was significantly less than that of E. haichowensis. Guard cells in E. haichowensis plants were covered with a thick cuticle layer, the epidermal hair was more numerous and longer, and the number of xylem conduits in the root was small. (4 The transpiration rate and the stomatal conductance of O. glazioviana were both significantly lower than those of E. haichowensis, regardless of whether the plants were treated with Cu. Taken together, these results indicate that the differences in the structural characteristics between these two plant species, particularly in the characteristics related to plant transpiration, are important factors that govern whether plants acquire or exclude Cu.

Lansoprazole induces apoptosis of breast cancer cells through inhibition of intracellular proton extrusion

Energy Technology Data Exchange (ETDEWEB)

Zhang, Shangrong; Wang, Yifan; Li, Shu Jie, E-mail: shujieli@nankai.edu.cn

2014-06-13

Highlights: • Lansoprazole (LPZ) induces cell apoptosis in breast cancer cells. • LPZ markedly inhibits intracellular proton extrusion. • LPZ induces an increase in intracellular ATP level, lysosomal alkalinization and ROS accumulation. - Abstract: The increased glycolysis and proton secretion in tumors is proposed to contribute to the proliferation and invasion of cancer cells during the process of tumorigenesis and metastasis. Here, treatment of human breast cancer cells with proton pump inhibitor (PPI) lansoprazole (LPZ) induces cell apoptosis in a dose-dependent manner. In the implantation of the MDA-MB-231 xenografts in nude mice, administration of LPZ significantly inhibits tumorigenesis and induces large-scale apopotosis of tumor cells. LPZ markedly inhibits intracellular proton extrusion, induces an increase in intracellular ATP level, lysosomal alkalinization and accumulation of reactive oxygen species (ROS) in breast cancer cells. The ROS scavenger N-acetyl-L-cysteine (NAC) and diphenyleneiodonium (DPI), a specific pharmacological inhibitor of NADPH oxidases (NOX), significantly abolish LPZ-induced ROS accumulation in breast cancer cells. Our results suggested that LPZ may be used as a new therapeutic drug for breast tumor.

Lansoprazole induces apoptosis of breast cancer cells through inhibition of intracellular proton extrusion

International Nuclear Information System (INIS)

Zhang, Shangrong; Wang, Yifan; Li, Shu Jie

2014-01-01

Highlights: • Lansoprazole (LPZ) induces cell apoptosis in breast cancer cells. • LPZ markedly inhibits intracellular proton extrusion. • LPZ induces an increase in intracellular ATP level, lysosomal alkalinization and ROS accumulation. - Abstract: The increased glycolysis and proton secretion in tumors is proposed to contribute to the proliferation and invasion of cancer cells during the process of tumorigenesis and metastasis. Here, treatment of human breast cancer cells with proton pump inhibitor (PPI) lansoprazole (LPZ) induces cell apoptosis in a dose-dependent manner. In the implantation of the MDA-MB-231 xenografts in nude mice, administration of LPZ significantly inhibits tumorigenesis and induces large-scale apopotosis of tumor cells. LPZ markedly inhibits intracellular proton extrusion, induces an increase in intracellular ATP level, lysosomal alkalinization and accumulation of reactive oxygen species (ROS) in breast cancer cells. The ROS scavenger N-acetyl-L-cysteine (NAC) and diphenyleneiodonium (DPI), a specific pharmacological inhibitor of NADPH oxidases (NOX), significantly abolish LPZ-induced ROS accumulation in breast cancer cells. Our results suggested that LPZ may be used as a new therapeutic drug for breast tumor

Inhibition of the Mitochondrial Glutamate Carrier SLC25A22 in Astrocytes Leads to Intracellular Glutamate Accumulation

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Emmanuelle Goubert

2017-05-01

Full Text Available The solute carrier family 25 (SLC25 drives the import of a large diversity of metabolites into mitochondria, a key cellular structure involved in many metabolic functions. Mutations of the mitochondrial glutamate carrier SLC25A22 (also named GC1 have been identified in early epileptic encephalopathy (EEE and migrating partial seizures in infancy (MPSI but the pathophysiological mechanism of GC1 deficiency is still unknown, hampered by the absence of an in vivo model. This carrier is mainly expressed in astrocytes and is the principal gate for glutamate entry into mitochondria. A sufficient supply of energy is essential for the proper function of the brain and mitochondria have a pivotal role in maintaining energy homeostasis. In this work, we wanted to study the consequences of GC1 absence in an in vitro model in order to understand if glutamate catabolism and/or mitochondrial function could be affected. First, short hairpin RNA (shRNA designed to specifically silence GC1 were validated in rat C6 glioma cells. Silencing GC1 in C6 resulted in a reduction of the GC1 mRNA combined with a decrease of the mitochondrial glutamate carrier activity. Then, primary astrocyte cultures were prepared and transfected with shRNA-GC1 or mismatch-RNA (mmRNA constructs using the Neon® Transfection System in order to target a high number of primary astrocytes, more than 64%. Silencing GC1 in primary astrocytes resulted in a reduced nicotinamide adenine dinucleotide (Phosphate (NAD(PH formation upon glutamate stimulation. We also observed that the mitochondrial respiratory chain (MRC was functional after glucose stimulation but not activated by glutamate, resulting in a lower level of cellular adenosine triphosphate (ATP in silenced astrocytes compared to control cells. Moreover, GC1 inactivation resulted in an intracellular glutamate accumulation. Our results show that mitochondrial glutamate transport via GC1 is important in sustaining glutamate homeostasis in

Aging results in copper accumulations in glial fibrillary acidic protein-positive cells in the subventricular zone.

Science.gov (United States)

Pushkar, Yulia; Robison, Gregory; Sullivan, Brendan; Fu, Sherleen X; Kohne, Meghan; Jiang, Wendy; Rohr, Sven; Lai, Barry; Marcus, Matthew A; Zakharova, Taisiya; Zheng, Wei

2013-10-01

Analysis of rodent brains with X-ray fluorescence (XRF) microscopy combined with immunohistochemistry allowed us to demonstrate that local Cu concentrations are thousands of times higher in the glia of the subventricular zone (SVZ) than in other cells. Using XRF microscopy with subcellular resolution and intracellular X-ray absorption spectroscopy we determined the copper (I) oxidation state and the sulfur ligand environment. Cu K-edge X-ray absorption near edge spectroscopy is consistent with Cu being bound as a multimetallic Cu-S cluster similar to one present in Cu-metallothionein. Analysis of age-related changes show that Cu content in astrocytes of the SVZ increases fourfold from 3 weeks to 9 months, while Cu concentration in other brain areas remain essentially constant. This increase in Cu correlates with a decrease in adult neurogenesis assessed using the Ki67 marker (both, however, can be age-related effects). We demonstrate that the Cu distribution and age-related concentration changes in the brain are highly cell specific. © 2013 The Anatomical Society and John Wiley & Sons Ltd.

Investigating Internalization and Intracellular Trafficking of GPCRs

DEFF Research Database (Denmark)

Foster, Simon R; Bräuner-Osborne, Hans

2017-01-01

for signal transduction. One of the major mechanisms for GPCR regulation involves their endocytic trafficking, which serves to internalize the receptors from the plasma membrane and thereby attenuate G protein-dependent signaling. However, there is accumulating evidence to suggest that GPCRs can signal...... independently of G proteins, as well as from intracellular compartments including endosomes. It is in this context that receptor internalization and intracellular trafficking have attracted renewed interest within the GPCR field. In this chapter, we will review the current understanding and methodologies...

Correlation between oxalic acid production and copper tolerance in Wolfiporia cocos

Science.gov (United States)

C. A. Clausen; Frederick Green; B. M. Woodward; J. W. Evans; R. C. DeGroot

2000-01-01

The increased interest in copper-based wood preservatives has hastened the need for understanding why some fungi are able to attack copper-treated wood. Due in part to accumulation of oxalic acid by brown-rot fungi and visualization of copper oxalate crystals in wood decayed by known copper-tolerant decay fungi, oxalic acid has been implicated in copper tolerance by...

ROS dependent copper toxicity in Hydra-biochemical and molecular study.

Science.gov (United States)

Zeeshan, Mohammed; Murugadas, Anbazhagan; Ghaskadbi, Surendra; Rajendran, Ramasamy Babu; Akbarsha, Mohammad Abdulkader

2016-01-01

Copper, an essential microelement, is known to be toxic to aquatic life at concentrations higher than that could be tolerated. Copper-induced oxidative stress has been documented in vitro, yet the in vivo effects of metal-induced oxidative stress have not been extensively studied in the lower invertebrates. The objective of the present study has been to find the effect of ROS-mediated toxicity of environmentally relevant concentrations of copper at organismal and cellular levels in Hydra magnipapillata. Exposure to copper at sublethal concentrations (0.06 and 0.1mg/L) for 24 or 48h resulted in generation of significant levels of intracellular reactive oxygen species (ROS). We infer that the free radicals here originate predominantly at the lysosomes but partly at the mitochondria also as visualized by H2-DHCFDA staining. Quantitative real-time PCR of RNA extracted from copper-exposed polyps revealed dose-dependent up-regulation of all antioxidant response genes (CAT, SOD, GPx, GST, GR, G6PD). Concurrent increase of Hsp70 and FoxO genes suggests the ability of polyps to respond to stress, which at 48h was not the same as at 24h. Interestingly, the transcript levels of all genes were down-regulated at 48h as compared to 24h incubation period. Comet assay indicated copper as a powerful genotoxicant, and the DNA damage was dose- as well as duration-dependent. Western blotting of proteins (Bax, Bcl-2 and caspase-3) confirmed ROS-mediated mitochondrial cell death in copper-exposed animals. These changes correlated well with changes in morphology, regeneration and aspects of reproduction. Taken together, the results indicate increased production of intracellular ROS in Hydra on copper exposure. Copyright © 2016 Elsevier Inc. All rights reserved.

Nutritional managment of inherited copper-associated hepatitis in the Labrador retriever.

NARCIS (Netherlands)

Fieten, Hille; Biourge, Vincent; Watson, AL; Leegwater, Peter; van den Ingh, T.S.G.A.M.; Rothuizen, J

Canine hereditary copper-associated hepatitis is characterized by gradual hepatic copper accumulation eventually leading to liver cirrhosis. Therapy is aimed at creating a negative copper balance with metal chelators, of which d-penicillamine is the most commonly used. d-penicillamine often causes

Copper localization, elemental content, and thallus colour in the copper hyperaccumulator lichen Lecanora sierra from California

Science.gov (United States)

Purvis, O.W.; Bennett, J.P.; Spratt, J.

2011-01-01

An unusual dark blue-green lichen, Lecanora sierrae, was discovered over 30 years ago by Czehura near copper mines in the Lights Creek District, Plumas County, Northern California. Using atomic absorption spectroscopy, Czehura found that dark green lichen samples from Warren Canyon contained 4% Cu in ash and suggested that its colour was due to copper accumulation in the cortex. The present study addressed the hypothesis that the green colour in similar material we sampled from Warren Canyon in 2008, is caused by copper localization in the thallus. Optical microscopy and electron microprobe analysis of specimens of L. sierrae confirmed that copper localization took place in the cortex. Elemental analyses of L. sierrae and three other species from the same localities showed high enrichments of copper and selenium, suggesting that copper selenates or selenites might occur in these lichens and be responsible for the unusual colour.

Copper localization, elemental content, and thallus colour in the copper hyperaccumulator lichen Lecanora sierrae from California

Science.gov (United States)

Purvis, O.W.; Bennett, J.P.; Spratt, J.

2011-01-01

An unusual dark blue-green lichen, Lecanora sierrae, was discovered over 30 years ago by Czehura near copper mines in the Lights Creek District, Plumas County, Northern California. Using atomic absorption spectroscopy, Czehura found that dark green lichen samples from Warren Canyon contained 4% Cu in ash and suggested that its colour was due to copper accumulation in the cortex. The present study addressed the hypothesis that the green colour in similar material we sampled from Warren Canyon in 2008, is caused by copper localization in the thallus. Optical microscopy and electron microprobe analysis of specimens of L. sierrae confirmed that copper localization took place in the cortex. Elemental analyses of L. sierrae and three other species from the same localities showed high enrichments of copper and selenium, suggesting that copper selenates or selenites might occur in these lichens and be responsible for the unusual colour. Copyright ?? 2011 British Lichen Society.

Higher biomolecules yield in phytoplankton under copper exposure.

Science.gov (United States)

Silva, Jaqueline Carmo; Echeveste, Pedro; Lombardi, Ana Teresa

2018-05-30

Copper is an important metal for industry, and its toxic threshold in natural ecosystems has increased since the industrial revolution. As an essential nutrient, it is required in minute amounts, being toxic in slightly increased concentrations, causing great biochemical transformation in microalgae. This study aimed at investigating the physiology of Scenedesmus quadricauda, a cosmopolitan species, exposed to copper concentrations including those that trigger intracellular biochemical modifications. The Cu exposure concentrations tested ranged from 0.1 to 25 µM, thus including environmentally important levels. Microalgae cultures were kept under controlled environmental conditions and monitored daily for cell density, in vivo chlorophyll a, and photosynthetic quantum yield (Φ M ). After 24 h growth, free Cu 2+ ions were determined, and after 96 h, cellular Cu concentration, total carbohydrates, proteins, lipids, and cell volume were determined. The results showed that both free Cu 2+ ions and cellular Cu increased with Cu increase in culture medium. Microalgae cell abundance and in vivo chlorophyll a were mostly affected at 2.5 µM Cu exposure (3.8 pg Cu cell -1 ) and above. Approximately 31% decrease of photosynthetic quantum yield was obtained at the highest Cu exposure concentration (25 µM; 25 pg Cu cell -1 ) in comparison with the control. However, at environmentally relevant copper concentrations (0.5 µM Cu; 0.4 pg Cu cell -1 ) cell volume increased in comparison with the control. Considering biomolecules accumulation per unit cell volume, the highest carbohydrates and proteins yield was obtained at 1.0 µM Cu (1.1 pg Cu cell -1 ), while for lipids higher Cu was necessary (2.5 µM Cu; 3.8 pg Cu cell -1 ). This study is a contribution to the understanding of the effects of environmentally significant copper concentrations in the physiology of S. quadricauda, as well as to biotechnological approach to increase biomolecule yield in

Intracellular distribution of nontargeted quantum dots after natural uptake and microinjection

Science.gov (United States)

Damalakiene, Leona; Karabanovas, Vitalijus; Bagdonas, Saulius; Valius, Mindaugas; Rotomskis, Ricardas

2013-01-01

Background: The purpose of this study was to elucidate the mechanism of natural uptake of nonfunctionalized quantum dots in comparison with microinjected quantum dots by focusing on their time-dependent accumulation and intracellular localization in different cell lines. Methods: The accumulation dynamics of nontargeted CdSe/ZnS carboxyl-coated quantum dots (emission peak 625 nm) was analyzed in NIH3T3, MCF-7, and HepG2 cells by applying the methods of confocal and steady-state fluorescence spectroscopy. Intracellular colocalization of the quantum dots was investigated by staining with Lysotracker®. Results: The uptake of quantum dots into cells was dramatically reduced at a low temperature (4°C), indicating that the process is energy-dependent. The uptake kinetics and imaging of intracellular localization of quantum dots revealed three accumulation stages of carboxyl-coated quantum dots at 37°C, ie, a plateau stage, growth stage, and a saturation stage, which comprised four morphological phases: adherence to the cell membrane; formation of granulated clusters spread throughout the cytoplasm; localization of granulated clusters in the perinuclear region; and formation of multivesicular body-like structures and their redistribution in the cytoplasm. Diverse quantum dots containing intracellular vesicles in the range of approximately 0.5–8 μm in diameter were observed in the cytoplasm, but none were found in the nucleus. Vesicles containing quantum dots formed multivesicular body-like structures in NIH3T3 cells after 24 hours of incubation, which were Lysotracker-negative in serum-free medium and Lysotracker-positive in complete medium. The microinjected quantum dots remained uniformly distributed in the cytosol for at least 24 hours. Conclusion: Natural uptake of quantum dots in cells occurs through three accumulation stages via a mechanism requiring energy. The sharp contrast of the intracellular distribution after microinjection of quantum dots in comparison

Accumulation of some metal ions on Bacillus licheniformis

International Nuclear Information System (INIS)

Hafez, M.B.; El-Desouky, W.; Fouad, A.

2001-01-01

Pure species of Bacillus licheniformis was used to remove ions from aqueous and simulated waste solutions. Metal ion accumulation on B. licheniformis was fast. Maximum uptake occurred at pH 4± 0.5 and at 25 ± 3 deg C. One gram of dry B. licheniformis was found to accumulate 115 mg cerium, 34 mg copper and 11 mg cobalt from aqueous solutions. The presence of certain foreign ions such as calcium, sodium and potassium decreased the uptake of ions by B. licheniformis, while citrate and EDTA prevent the uptake. Electron microscopic investigations showed that cerium (III), copper (II) and cobalt (II) accumulated extracellulary around the surface wall of B. licheniformis cells. A bio-adsorption mechanism between the metal ions and B. licheniformis cell wall was proposed. (author)

Characterization of Leptin Intracellular Trafficking

Directory of Open Access Journals (Sweden)

E Walum

2009-12-01

Full Text Available Leptin is produced by adipose tissue, and its concentration in plasma is related to the amount of fat in the body. The leptin receptor (OBR is a member of the class I cytokine receptor family and several different isoforms, produced by alternative mRNA splicing are found in many tissues, including the hypothalamus. The two predominant isoforms includes a long form (OBRl with an intracellular domain of 303 amino acids and a shorter form (OBRs with an intracellular domain of 34 amino acids. Since OBRl is mainly expressed in the hypotalamus, it has been suggested to be the main signalling form. The peripheral production of leptin by adipocyte tissue and its effects as a signal of satiety in the central nervous system imply that leptin gains access to regions of the brain regulating in energy balance by crossing the blood-brain barrier. In an attempt to characterize the intracellular transport of leptin, we have followed binding internalization and degradation of leptin in HEK293 cells. We have also monitored the intracellular transport pathway of fluorescent conjugated leptin in HEK293 cells. Phenylarsine oxide, a general inhibitor of endocytosis, as well as incubation at mild hypertonic conditions, prevented the uptake of leptin, confirming a receptor-mediated internalization process. When internalized, 125I-leptin was rapidly accumulated inside the cells and reached a maximum after 10 min. After 70 minutes about 40-50% of total counts in each time point were found in the medium as TCA-soluble material. Leptin sorting, at the level of early endosomes, did not seem to involve recycling endosomes, since FITC-leptin was sorted from Cy3- transferrin containing compartments at 37°C. At 45 minutes of continuos internalization, FITC-leptin appeared mainly accumulated in late endocytic structures colocalizing with internalized rhodamine coupled epidermial growth factor (EGF and the lysosomal marker protein lamp-1. The transport of leptin was also shown

Colchicine induced intraneuronal free zinc accumulation and dentate granule cell degeneration.

Science.gov (United States)

Choi, Bo Young; Lee, Bo Eun; Kim, Jin Hee; Kim, Hyun Jung; Sohn, Min; Song, Hong Ki; Chung, Tae Nyoung; Suh, Sang Won

2014-08-01

Colchicine has been discovered to inhibit many inflammatory processes such as gout, familial Mediterranean fever, pericarditis and Behcet disease. Other than these beneficial anti-inflammatory effects, colchicine blocks microtubule-assisted axonal transport, which results in the selective loss of dentate granule cells of the hippocampus. The mechanism of the colchicine-induced dentate granule cell death and depletion of mossy fiber terminals still remains unclear. In the present study, we hypothesized that colchicine-induced dentate granule cell death may be caused by accumulation of labile intracellular zinc. 10 μg kg(-1) of colchicine was injected into the adult rat hippocampus and then brain sections were evaluated at 1 day or 1 week later. Neuronal cell death was evaluated by H&E staining or Fluoro-Jade B. Zinc accumulation and vesicular zinc were detected by N-(6-methoxy-8-quinolyl)-para-toluene sulfonamide (TSQ) staining. To test whether an extracellular zinc chelator can prevent this process, CaEDTA was injected into the hippocampus over a 5 min period with colchicine. To test whether other microtubule toxins also produce similar effects as colchicine, vincristine was injected into the hippocampus. The present study found that colchicine injection induced intracellular zinc accumulation in the dentate granule cells and depleted vesicular zinc from mossy fiber terminals. Injection of a zinc chelator, CaEDTA, did not block the zinc accumulation and neuronal death. Vincristine also produced intracellular zinc accumulation and neuronal death. These results suggest that colchicine-induced dentate granule cell death is caused by blocking axonal zinc flow and accumulation of intracellular labile zinc.

A copper-induced quinone degradation pathway provides protection against combined copper/quinone stress in Lactococcus lactis IL1403.

Science.gov (United States)

Mancini, Stefano; Abicht, Helge K; Gonskikh, Yulia; Solioz, Marc

2015-02-01

Quinones are ubiquitous in the environment. They occur naturally but are also in widespread use in human and industrial activities. Quinones alone are relatively benign to bacteria, but in combination with copper, they become toxic by a mechanism that leads to intracellular thiol depletion. Here, it was shown that the yahCD-yaiAB operon of Lactococcus lactis IL1403 provides resistance to combined copper/quinone stress. The operon is under the control of CopR, which also regulates expression of the copRZA copper resistance operon as well as other L. lactis genes. Expression of the yahCD-yaiAB operon is induced by copper but not by quinones. Two of the proteins encoded by the operon appear to play key roles in alleviating quinone/copper stress: YaiB is a flavoprotein that converts p-benzoquinones to less toxic hydroquinones, using reduced nicotinamide adenine dinucleotide phosphate (NADPH) as reductant; YaiA is a hydroquinone dioxygenase that converts hydroquinone putatively to 4-hydroxymuconic semialdehyde in an oxygen-consuming reaction. Hydroquinone and methylhydroquinone are both substrates of YaiA. Deletion of yaiB causes increased sensitivity of L. lactis to quinones and complete growth arrest under combined quinone and copper stress. Copper induction of the yahCD-yaiAB operon offers protection to copper/quinone toxicity and could provide a growth advantage to L. lactis in some environments. © 2014 John Wiley & Sons Ltd.

Complexation of intracellular cyanide by hydroxocobalamin using a human cellular model.

Science.gov (United States)

Astier, A; Baud, F J

1996-01-01

1. The rational for administering hydroxocobalamin (OHCbl) as an antidote to cyanide poisoning is based on the high affinity of CN ion for cobalt compounds. However, only few data are available on the influence of OHCbl on the intracellular cyanide pool. 2. In human fibroblasts incubated for 10 min with 500 microM of [14C] cyanide, the accumulation ratio was 25 at 37 degrees C (10.45 +/- 1.51 mM) and 11.9 at 4 degrees C. 3. Using the monoblastic U-937 cell line, a rapid uptake of radioactive cyanide was observed with a maximum accumulation ratio of 1.97 at 5 min. 4. A linear relationship between cyanide uptake by U-937 cells and cyanide concentration in incubation medium (10-500 microM; 5 min) was found suggesting a first order process (k = 0.25 min-1). 5. After incubation of fibroblasts with 500 microM of OHCbl, a 75% decrease of intracellular cyanide was observed, with concomittant formation of intracellular cyanocobalamin CNCbl (intracellular/extracellular ratio: 158). 6. These findings suggest that OHCbl is able to penetrate into heavily cyanide loaded cells and to complex cyanide to the non-toxic CNCbl form.

Effects of coal spoil amendment on heavy metal accumulation and physiological aspects of ryegrass (Lolium perenne L.) growing in copper mine tailings.

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Chu, Zhaoxia; Wang, Xingming; Wang, Yunmin; Liu, Guijian; Dong, Zhongbing; Lu, Xianwen; Chen, Guangzhou; Zha, Fugeng

2017-12-21

Copper mine tailings pose many threats to the surrounding environment and human health, and thus, their remediation is fundamental. Coal spoil is the waste by-product of coal mining and characterized by low levels of metals, high content of organic matter, and many essential microelements. This study was designed to evaluate the role of coal spoil on heavy uptake and physiological responses of Lolium perenne L. grown in copper mine tailings amended with coal spoil at rates of 0, 0.5, 1, 5, 10, and 20%. The results showed that applying coal spoil to copper mine tailings decreased the diethylenetriaminepentaacetic acid (DTPA)-extractable Cd, Cu, Pb, and Zn contents in tailings and reduced those metal contents in both roots and shoots of the plant. However, application of coal spoil increased the DTPA-extractable Cr concentration in tailings and also increased Cr uptake and accumulation by Lolium perenne L. The statistical analysis of physiological parameters indicated that chlorophyll and carotenoid increased at the lower amendments of coal spoil followed by a decrease compared to their respective controls. Protein content was enhanced at all the coal spoil amendments. When treated with coal spoil, the activities of superoxide dismutases (SOD), peroxidase (POD), and catalase (CAT) responded differently. CAT activity was inhibited, but POD activity was increased with increasing amendment ratio of coal spoil. SOD activity increased up to 1% coal spoil followed by a decrease. Overall, the addition of coal spoil decreased the oxidative stress in Lolium perenne L., reflected by the reduction in malondialdehyde (MDA) contents in the plant. It is concluded that coal spoil has the potential to stabilize most metals studied in copper mine tailings and ameliorate the harmful effects in Lolium perenne L. through changing the physiological attributes of the plant grown in copper mine tailings.

Intracellular sodium hydrogen exchange inhibition and clinical myocardial protection.

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Mentzer, Robert M; Lasley, Robert D; Jessel, Andreas; Karmazyn, Morris

2003-02-01

Although the mechanisms underlying ischemia/reperfusion injury remain elusive, evidence supports the etiologic role of intracellular calcium overload and oxidative stress induced by reactive oxygen species. Activation of the sodium hydrogen exchanger (NHE) is associated with intracellular calcium accumulation. Inhibition of the NHE-1 isoform may attenuate the consequences of this injury. Although there is strong preclinical and early clinical evidence that NHE inhibitors may be cardioprotective, definitive proof of this concept in humans awaits the results of ongoing clinical trials.

Chronic copper poisoning. III. Effects of copper acetate injected into the bloodstream of sheep

Energy Technology Data Exchange (ETDEWEB)

Todd, J R; Thompson, R H

1964-01-01

A study was made of the clinical and biochemical effects of injections of copper (as acetate) into the bloodstream of sheep of 100 to 130 lb. liveweight. Copper in a dose of 160 mg. caused death in 3 sheep in a few hours, and 80 mg. caused death in 3 out of 4 sheep, 2 after 2 days and 1 after 11 days. Symptoms, biochemical lesions and post-mortem appearances did not resemble those of chronic copper poisoning, but rather those of gastro-enteritis. Blood glutathione concentrations were not markedly reduced, but haemoconcentration was a prominent feature. Post-mortem examination showed gross congestion of blood vessels and marked inflammatory reactions in the abomasum and small intestine. Single injections of smaller amounts (25 to 40 mg. copper) were tolerated without effect, but repeated injections, twice daily for 2 to 3 days, caused haemolytic episodes in 3 sheep similar to the crisis of chronic copper poisoning in that a marked reduction in blood glutathione concentration and accumulation of methaemoglobin occurred. No other clinical effects were produced, however, and all three animals recovered uneventfully.

Distribution and chemical forms of copper in the root cells of castor seedlings and their tolerance to copper phytotoxicity in hydroponic culture.

Science.gov (United States)

Kang, Wei; Bao, Jianguo; Zheng, Jin; Hu, Hongqin; Du, Jiangkun

2015-05-01

The subcellular localization and chemical forms of copper in castor (Ricinus communis L.) seedlings grown in hydroponic nutrient solution were identified by chemical extraction, transmission electron microscopy, and Fourier transform infrared spectroscopy. The wild castor seeds were harvested from an abandoned copper mine in Tonglu Mountain, Daye City of Hubei Province, China. The results revealed that (1) the seedlings grew naturally in MS liquid medium with 40.00 mg kg(-1) CuSO4, in which the seedling growth rate and biomass index were 0.14 and 1.23, respectively, which were the highest values among all the treatments. The copper content in castor seedlings increased along with elevated CuSO4 concentration in the medium, reaching a maximum value of 16 570.12 mg kg(-1)(DW) when exposed to 60.00 mg L(-1) CuSO4, where 91.31% of the copper was accumulated in roots. (2) The copper existed in various chemical forms in the roots of the castor seedlings. Copper of 67.66% was extracted from the components of cell walls, such as exchangeable acidic polar compounds, cellulose and lignin, protein and pectin, and less concentrated in cell cytoplasm and nuclei. (3) Furthermore, the root cell walls were thickened when the castor seedlings exposed to CuSO4, with a large amount of high-density electron bodies, attached to the thickened cell walls. In the cell walls, most copper was bound to the carboxyl (-COOH) and hydroxyl (-OH) groups of acidic polar compounds, cellulose, hemicellulose, and polysaccharides. The conclusion showed that castor exhibited a strong tolerance to copper, the copper were accumulated mainly in the root cell, the root cell walls of castor were the major location of patience and detoxification in copper stress.

Ligand-Doped Copper Oxo-hydroxide Nanoparticles are Effective Antimicrobials

Science.gov (United States)

Bastos, Carlos A. P.; Faria, Nuno; Ivask, Angela; Bondarenko, Olesja M.; Kahru, Anne; Powell, Jonathan

2018-04-01

Bacterial resistance to antimicrobial therapies is an increasing clinical problem. This is as true for topical applications as it is for systemic therapy. Topically, copper ions may be effective and cheap antimicrobials that act through multiple pathways thereby limiting opportunities to bacteria for resistance. However, the chemistry of copper does not lend itself to facile formulations that will readily release copper ions at biologically compatible pHs. Here, we have developed nanoparticulate copper hydroxide adipate tartrate (CHAT) as a cheap, safe, and readily synthesised material that should enable antimicrobial copper ion release in an infected wound environment. First, we synthesised CHAT and showed that this had disperse aquated particle sizes of 2-5 nm and a mean zeta potential of - 40 mV. Next, when diluted into bacterial medium, CHAT demonstrated similar efficacy to copper chloride against Escherichia coli and Staphylococcus aureus, with dose-dependent activity occurring mostly around 12.5-50 mg/L of copper. Indeed, at these levels, CHAT very rapidly dissolved and, as confirmed by a bacterial copper biosensor, showed identical intracellular loading to copper ions derived from copper chloride. However, when formulated at 250 mg/L in a topically applied matrix, namely hydroxyethyl cellulose, the benefit of CHAT over copper chloride was apparent. The former yielded rapid sustained release of copper within the bactericidal range, but the copper chloride, which formed insoluble precipitates at such concentration and pH, achieved a maximum release of 10 ± 7 mg/L copper by 24 h. We provide a practical formulation for topical copper-based antimicrobial therapy. Further studies, especially in vivo, are merited.

Copper Homeostasis in Escherichia coli and Other Enterobacteriaceae.

Science.gov (United States)

Rensing, Christopher; Franke, Sylvia

2007-04-01

An interesting model for studying environmental influences shaping microbial evolution is provided by a multitude of copper resistance and copper homeostasis determinants in enteric bacteria. This review describes these determinants and tries to relate their presence to the habitat of the respective organism, as a current hypothesis predicts that the environment should determine an organism's genetic makeup. In Escherichia coli there are four regulons that are induced in the presence of copper. Two, the CueR and the CusR regulons, are described in detail. A central component regulating intracellular copper levels, present in all free-living enteric bacteria whose genomes have so far been sequenced, is a Cu(I)translocating P-type ATPase. The P-type ATPase superfamily is a ubiquitous group of proteins involved in the transport of charged substrates across biological membranes. Whereas some components involved in copper homeostasis can be found in both anaerobes and aerobes, multi-copper oxidases (MCOs) implicated in copper tolerance in E. coli, such as CueO and the plasmid-based PcoA, can be found only in aerobic organisms. Several features indicate that CueO, PcoA, and other related MCOs are specifically adapted to combat copper-mediated oxidative damage. In addition to these well-characterized resistance operons, there are numerous other genes that appear to be involved in copper binding and trafficking that have not been studied in great detail. SilE and its homologue PcoE, for example, are thought to effect the periplasmic binding and sequestration of silver and copper, respectively.

Tolerance to and Accumulation of Cadmium, Copper, and Zinc by Cupriavidus necator

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Rayssa Pereira Vicentin

2018-03-01

Full Text Available ABSTRACT Preliminary results of in vitro experiments with multicontaminated soils and solid media indicated that nodulating diazotrophic bacteria of the genus Cupriavidus are promising for the remediation of contaminated environments due to their symbiosis with legumes and metal tolerance. Thus, strains of Cupriavidus spp. (LMG 19424T, UFLA 01-659, UFLA 01-663, and UFLA 02-71 were tested for their ability to tolerate and bioaccumulate cadmium (Cd, copper (Cu, and zinc (Zn in Luria-Bertani broth. Changes in the growth pattern of Cupriavidus strains in the presence or absence of heavy metals were analyzed by scanning electron microscopy and metal allocation by transmission electron microscopy, to clarify the mechanisms of bioremediation. Highest tolerance was detected for strain UFLA 01-659 (minimum inhibitory concentration of 5, 4.95, and 14.66 mmol L−1 of Cd, Cu, and Zn, respectively. Among the removal rates of the metals tested (9.0, 4.6, and 3.2 mg L−1 of Cd, Cu, and Zn, respectively, the bacterial activity was clearly highest for Cd. The efficiency of strain UFLA 01-659 in removing the heavy metals is associated with its high biomass production and/or higher contents of heavy metals adsorbed and absorbed in the biomass. In response to the presence of heavy metals in the liquid culture medium, the bacteria produced exopolysaccharides and small and aggregated cells. However, these responses varied according to the strains and heavy metals. Regarding allocation, all heavy metals were adsorbed on the cell wall and membrane, whereas complexation was observed intracellularly and only for Cu and Zn. These results indicate the possibility of using C. necator UFLA 01-659 for remediation in areas with very high Cd, Cu, and Zn contents.

Two Outer Membrane Proteins Contribute to Caulobacter crescentus Cellular Fitness by Preventing Intracellular S-Layer Protein Accumulation

Energy Technology Data Exchange (ETDEWEB)

Overton, K. Wesley; Park, Dan M.; Yung, Mimi C.; Dohnalkova, Alice; Smit, John; Jiao, Yongqin

2016-09-23

Surface layers, or S-layers, are two-dimensional protein arrays that form the outermost layer of many bacteria and archaea. They serve several functions, including physical protection of the cell from environmental threats. The high abundance of S-layer proteins necessitates a highly efficient export mechanism to transport the S-layer protein from the cytoplasm to the cell exterior.Caulobacter crescentusis unique in that it has two homologous, seemingly redundant outer membrane proteins, RsaF_aand RsaF_b, which together with other components form a type I protein translocation pathway for S-layer export. These proteins have homology toEscherichia coliTolC, the outer membrane channel of multidrug efflux pumps. Here we provide evidence that, unlike TolC, RsaF_aand RsaF_bare not involved in either the maintenance of membrane stability or the active export of antimicrobial compounds. Rather, RsaF_aand RsaF_bare required to prevent intracellular accumulation and aggregation of the S-layer protein RsaA; deletion of RsaF_aand RsaF_bled to a general growth defect and lowered cellular fitness. Using Western blotting, transmission electron microscopy, and transcriptome sequencing (RNA-seq), we show that loss of both RsaF_aand RsaF_bled to accumulation of insoluble RsaA in the cytoplasm, which in turn caused upregulation of a number of genes involved in protein misfolding and degradation pathways. These findings provide new insight into the requirement for RsaF_aand RsaF_bin cellular fitness and tolerance to antimicrobial agents and further our understanding of the S-layer export mechanism on both the transcriptional and translational levels in

Bioaccumulation and Depuration of Copper in the Kidney and Liver of a Freshwater Fish, Capoeta fusca

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Borhan Mansouri

2016-07-01

Full Text Available Background: This study aims to investigate the patterns of bioaccumulation and depuration of copper in the selected kidney and liver of Capoeta fusca. Methods: The fish were collected between September and November 2010 from a qanat in Birjand. They were exposed to two types treatments with copper (0.25 and 0.75 mg/L for a period of 41 days. The fish under study were exposed to the above-mentioned sub-lethal concentrations separately for 14 and 21 days (accumulation period. At the end of this period, the remaining fish were kept in tap water (elimination period for 31 and 41 days. Results: The findings showed that the accumulation of copper in lower and higher sub-lethal concentrations was higher in kidney as the mean accumulation of copper on day 21 was 1.9±0.1 μg/g and 2.93±0.47 μg/g respectively, in 0.25 μg/g and 0.75 μg/g concentrations. On the other hand, the results also showed that the depuration level of copper in the given concentrations was higher in liver than kidney. The bioaccumulation and depuration of copper significantly increased in the kidney and liver of C. fusca (P<0.01. Conclusion: Based on the present work, it is concluded that C. fusca has a potential for the rapid accumulation and depuration of copper in freshwater. Also, the results indicate that the fish C. fusca, as representative fish species in the East of Iran, can be a useful bioindicator organism of water contamination with copper.

Determination of copper in whole blood by differential pulse adsorptive stripping voltammetry

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Tarik Attar

2014-02-01

Full Text Available A selective and sensitive method for determination of copper in blood by adsorptive differential pulse cathodic stripping voltammetry is presented. The method is based on adsorptive accumulation of the complexes of Cu (II ions with benzenesulfonyl hydrazide onto hanging mercury drop electrode (HMDE, followed by the reduction of the adsorbed species by differential pulse cathodic stripping voltammetry. The effect of various parameters such as supporting electrolyte, concentration of benzenesulfonyl hydrazide, accumulation potential, accumulation time and stirring rate on the selectivity and sensitivity were studied. The optimum conditions for determination of copper include perchloric acid 0.03 M, concentration of benzenesulfonyl hydrazide 7.5×10-5 M, the accumulation potential of -350 mV (vs. Ag/AgCl, the accumulation time of 50 s, and the scan rate of 50 mV s-1. Under optimized conditions, linear calibration curves were established for the concentration of Cu (II in the range of 0.62-275 ng mL-1, with detection limit of 0.186 ng mL-1 for Cu (II. The procedure was successfully applied to the determination of copper ion in whole blood samples.

Copper in the sea: a bibliography

Energy Technology Data Exchange (ETDEWEB)

Schmidt, R.L.

1977-04-01

Life in the sea is vulnerable to the influx of trace metals resulting from man's activities. Although many pollutants introduced to the sea eventually degrade to less harmful forms, trace metals accumulate in sediments and have a continued potential for effect on biota. Copper has a toxic potential exceeding all other metals due to the quantity discharged and its toxicological effect. Fortunately, copper in the oceans is rendered less bioavailable or less toxic by its ready interaction with the complex chemical components of seawater. This bibliography was prepared to illustrate the status of current knowledge of the biogeochemistry of copper and to aid the development of research programs to define the effects of copper discharged to the marine environment. The references are categorized to aid the reader to locate literature concerning specific aspects of the biogeochemistry of copper. A brief comment describing the important findings in each category is given. Although this bibliography is not exhaustive, the listed references are likely representative of current knowledge.

Arylthiazole antibiotics targeting intracellular methicillin-resistant Staphylococcus aureus (MRSA) that interfere with bacterial cell wall synthesis.

Science.gov (United States)

Eid, Islam; Elsebaei, Mohamed M; Mohammad, Haroon; Hagras, Mohamed; Peters, Christine E; Hegazy, Youssef A; Cooper, Bruce; Pogliano, Joe; Pogliano, Kit; Abulkhair, Hamada S; Seleem, Mohamed N; Mayhoub, Abdelrahman S

2017-10-20

The promising antibacterial potency of arylthiazole antibiotics is offset by their limited activity against intracellular bacteria (namely methicillin-resistant Staphylococcus aureus (MRSA)), similar to many clinically-approved antibiotics. The failure to target these hidden pathogens is due to the compounds' lack of proper characteristics to accumulate intracellularly. Fine tuning of the size and polar-surface-area of the linking heteroaromatic ring provided a new series of 5-thiazolylarylthiazoles with balanced properties that allow them to sufficiently cross and accumulate inside macrophages infected with MRSA. The most promising compound 4i exhibited rapid bactericidal activity, good metabolic stability and produced over 80% reduction of intracellular MRSA in infected macrophages. Copyright © 2017 Elsevier Masson SAS. All rights reserved.

Influence of intracellular Na + , K + and Cl - on the salt tolerance in ...

African Journals Online (AJOL)

In the process of selection for salt tolerance, it is important to understand the physiological basis of ion management executed by the cells through the exclusion, accumulation or maintenance of ratios of specific ions. Intracellular accumulation of Na+, K+ and Cl- ions in the cells in vitro was studied as a factor in salt ...

An Investigation of Low Biofouling Copper-charged Membranes

Science.gov (United States)

Asapu, Sunitha

Water is essential for the survival of life on Earth, but pollutants in water can cause dangerous diseases and fatalities. The need for purified water has been increasing with increasing world population; however, natural sources of water such as rivers, lakes and streams, are progressively falling shorter and shorter of meeting water needs. The provision of clean, drinkable water to people is a key factor for the development of novel and alternative water purification technologies, such as membrane separations. Nanofiltration (NF) is a membrane separations technology that purifies water from lower quality sources, such as brackish water, seawater and wastewater. During the filtration of such sources, materials that are rejected by the membrane may accumulate on the surface of the membrane to foul it. Such materials include organic and inorganic matter, colloids, salts and microorganisms. The former four can often be controlled via pretreatment; however, the accumulation of microorganisms is more problematic to membranes. Biofouling is the accumulation and growth of microorganisms on the surface of membranes and on feed spacers. After attachment, microorganisms excrete extracellular polymeric substances (EPS), which form a matrix around the organism's outer surface as biofilm. These biofilms are detrimental and result in irreversible membrane fouling. Copper and silver ions inactivate the bacterial cells and prevent the DNA replication in microbial cells. Previous studies using copper-charged feed spacers have shown the ability of copper to control biofouling without a significant amount of copper leaching from copper-charged polypropylene (PP) feed spacers during crossflow filtration. Also, filtration using unmodified speed facers experienced almost 70% flux decline, while filtration using copper-charged feed spacers displayed only 25% flux decline. These intriguing results led to the hypothesis that the polymer chemistry could be extrapolated to produce membranes

Intracellular cholesterol level regulates sensitivity of glioblastoma cells against temozolomide-induced cell death by modulation of caspase-8 activation via death receptor 5-accumulation and activation in the plasma membrane lipid raft.

Science.gov (United States)

Yamamoto, Yutaro; Tomiyama, Arata; Sasaki, Nobuyoshi; Yamaguchi, Hideki; Shirakihara, Takuya; Nakashima, Katsuhiko; Kumagai, Kosuke; Takeuchi, Satoru; Toyooka, Terushige; Otani, Naoki; Wada, Kojiro; Narita, Yoshitaka; Ichimura, Koichi; Sakai, Ryuichi; Namba, Hiroki; Mori, Kentaro

2018-01-01

Development of resistance against temozolomide (TMZ) in glioblastoma (GBM) after continuous treatment with TMZ is one of the critical problems in clinical GBM therapy. Intracellular cholesterol regulates cancer cell biology, but whether intracellular cholesterol is involved in TMZ resistance of GBM cells remains unclear. The involvement of intracellular cholesterol in acquired resistance against TMZ in GBM cells was investigated. Intracellular cholesterol levels were measured in human U251 MG cells with acquired TMZ resistance (U251-R cells) and TMZ-sensitive control U251 MG cells (U251-Con cells), and found that the intracellular cholesterol level was significantly lower in U251-R cells than in U251-Con cells. In addition, treatment by intracellular cholesterol remover, methyl-beta cyclodextrin (MβCD), or intracellular cholesterol inducer, soluble cholesterol (Chol), regulated TMZ-induced U251-Con cell death in line with changes in intracellular cholesterol level. Involvement of death receptor 5 (DR5), a death receptor localized in the plasma membrane, was evaluated. TMZ without or with MβCD and/or Chol caused accumulation of DR5 into the plasma membrane lipid raft and formed a complex with caspase-8, an extrinsic caspase cascade inducer, reflected in the induction of cell death. In addition, treatment with caspase-8 inhibitor or knockdown of DR5 dramatically suppressed U251-Con cell death induced by combination treatment with TMZ, MβCD, and Chol. Combined treatment of Chol with TMZ reversed the TMZ resistance of U251-R cells and another GBM cell model with acquired TMZ resistance, whereas clinical antihypercholesterolemia agents at physiological concentrations suppressed TMZ-induced cell death of U251-Con cells. These findings suggest that intracellular cholesterol level affects TMZ treatment of GBM mediated via a DR5-caspase-8 mechanism. Copyright © 2017 Elsevier Inc. All rights reserved.

Enhanced intracellular delivery and antibacterial efficacy of enrofloxacin-loaded docosanoic acid solid lipid nanoparticles against intracellular Salmonella.

Science.gov (United States)

Xie, Shuyu; Yang, Fei; Tao, Yanfei; Chen, Dongmei; Qu, Wei; Huang, Lingli; Liu, Zhenli; Pan, Yuanhu; Yuan, Zonghui

2017-01-23

Enrofloxacin-loaded docosanoic acid solid lipid nanoparticles (SLNs) with different physicochemical properties were developed to enhance activity against intracellular Salmonella. Their cellular uptake, intracellular elimination and antibacterial activity were studied in RAW 264.7 cells. During the experimental period, SLN-encapsulated enrofloxacin accumulated in the cells approximately 27.06-37.71 times more efficiently than free drugs at the same extracellular concentration. After incubation for 0.5 h, the intracellular enrofloxacin was enhanced from 0.336 to 1.147 μg/mg of protein as the sizes of nanoparticles were increased from 150 to 605 nm, and from 0.960 to 1.147 μg/mg of protein when the charge was improved from -8.1 to -24.9 mv. The cellular uptake was more significantly influenced by the size than it was by the charge, and was not affected by whether the charge was positive or negative. The elimination of optimal SLN-encapsulated enrofloxacin from the cells was significantly slower than that of free enrofloxacin after removing extracellular drug. The inhibition effect against intracellular Salmonella CVCC541 of 0.24 and 0.06 μg/mL encapsulated enrofloxacin was stronger than 0.6 μg/mL free drug after all of the incubation periods and at 48 h, respectively. Docosanoic acid SLNs are thus considered as a promising carrier for intracellular bacterial treatment.

Multiple Degradation Pathways of Chemoattractant Mediated Cyclic GMP Accumulation in Dictyostelium

NARCIS (Netherlands)

Haastert, Peter J.M. van; Lookeren Campagne, Michiel M. van; Kesbeke, Fanja

1983-01-01

Chemoattractants induce a transient accumulation of cGMP levels in Dictyostelium. Intracellular cGMP levels reach a peak at 10 s and prestimulated cGMP levels are recovered at about 30 s. Intracellular and extracellular cGMP levels were detected simultaneously after stimulation of D. lacteum cells

Curcumin Mitigates the Intracellular Lipid Deposit Induced by Antipsychotics In Vitro.

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Alberto Canfrán-Duque

Full Text Available First- and second-generation antipsychotics (FGAs and SGAs, respectively, both inhibit cholesterol biosynthesis and impair the intracellular cholesterol trafficking, leading to lipid accumulation in the late endosome/lysosome compartment. In this study we examined if curcumin, a plant polyphenol that stimulates exosome release, can alleviate antipsychotic-induced intracellular lipid accumulation.HepG2 hepatocarcinoma cells were treated with antipsychotics or placebo and DiI-labelled LDL for 18 h and then exposed to curcumin for the last 2 h. Cells and media were collected separately and used for biochemical analyses, electron microscopy and immunocytochemistry. Exosomes were isolated from the incubation medium by ultracentrifugation.Curcumin treatment reduced the number of heterolysosomes and shifted their subcellular localization to the periphery, as revealed by electron microscopy, and stimulated the release of lysosomal β-hexosaminidase and exosome markers flotillin-2 and CD63 into the media. The presence of DiI in exosomes released by cells preloaded with DiI-LDL demonstrated the endolysosomal origin of the microvesicles. Furthermore, curcumin increased the secretion of cholesterol as well as LDL-derived DiI and [3H]-cholesterol, in association with a decrease of intracellular lipids. Thus, the disruption of lipid trafficking induced by FGAs or SGAs can be relieved by curcumin treatment. This polyphenol, however, did not mitigate the reduction of cholesterol esterification induced by antipsychotics.Curcumin stimulates exosome release to remove cholesterol (and presumably other lipids accumulated within the endolysosomal compartment, thereby normalizing intracellular lipid homeostasis. This action may help minimize the adverse metabolic effects of antipsychotic treatment, which should now be evaluated in clinical trials.

Accumulation and Distribution of Lead and Copper in Avicennia marina and Rhizophora apiculata from Balok Mangrove Forest, Pahang, Malaysia

International Nuclear Information System (INIS)

Kamaruzzaman, B.Y.; Sharlinda, M.Z.R.; John, B.A.; Waznah, A.S.

2011-01-01

Bioaccumulation of lead and Copper in Avicennia marina and Rhizophora apiculata was studied. Samples of leaves, barks and roots were collected from Balok mangrove forest, Pahang. Pb and Cu accumulation was higher in Avicennia marina root tissue compared to bark and leaf but lower than surrounding sediment level. The average concentration of Pb in A. marina leaf, bark, root and sediment was observed to be 5.39 ppm, 3.63 ppm, 18.21 ppm and 23.13 ppm, and average Cu concentration was 4.13 ppm, 4.27 ppm, 4.81 ppm and 12.33 ppm, respectively. R. apiculata also showed higher concentration of Pb and Cu in root tissue compared to bark and leaf tissues but lower than surrounding sediment. The average concentration of Pb in R. apiculata leaf, bark, root and sediment was observed to be 4.30 ppm, 2.97 ppm, 22.45 ppm and 31.23 ppm, respectively. The average Cu concentration was 2.93 ppm, 4.71 ppm, 4.81 ppm and 15.52 ppm, respectively. Results of concentration factors (CF) showed that the accumulation of Pb and Cu was higher in A. marina than in R. apiculata. (author)

Effects of intracellular iron overload on cell death and identification of potent cell death inhibitors.

Science.gov (United States)

Fang, Shenglin; Yu, Xiaonan; Ding, Haoxuan; Han, Jianan; Feng, Jie

2018-06-11

Iron overload causes many diseases, while the underlying etiologies of these diseases are unclear. Cell death processes including apoptosis, necroptosis, cyclophilin D-(CypD)-dependent necrosis and a recently described additional form of regulated cell death called ferroptosis, are dependent on iron or iron-dependent reactive oxygen species (ROS). However, whether the accumulation of intracellular iron itself induces ferroptosis or other forms of cell death is largely elusive. In present study, we study the role of intracellular iron overload itself-induced cell death mechanisms by using ferric ammonium citrate (FAC) and a membrane-permeable Ferric 8-hydroxyquinoline complex (Fe-8HQ) respectively. We show that FAC-induced intracellular iron overload causes ferroptosis. We also identify 3-phosphoinositide-dependent kinase 1 (PDK1) inhibitor GSK2334470 as a potent ferroptosis inhibitor. Whereas, Fe-8HQ-induced intracellular iron overload causes unregulated necrosis, but partially activates PARP-1 dependent parthanatos. Interestingly, we identify many phenolic compounds as potent inhibitors of Fe-8HQ-induced cell death. In conclusion, intracellular iron overload-induced cell death form might be dependent on the intracellular iron accumulation rate, newly identified cell death inhibitors in our study that target ferroptosis and unregulated oxidative cell death represent potential therapeutic strategies against iron overload related diseases. Copyright © 2018 Elsevier Inc. All rights reserved.

Cloning and expression analysis of a blue copper- binding protein ...

African Journals Online (AJOL)

Jane

2011-07-20

Jul 20, 2011 ... decreased to constitutive level at 72 h after inoculation in resistant Gh21 line ... lignification of cell wall or scavenging of reactive oxygen species (ROS) during powdery mildew attack ... or other ions in plants (Lin and Wu, 1994), whose .... nutrient-uptake and copper accumulation in protein of copper-tolerant.

Subcellular site and nature of intracellular cadmium in plants

International Nuclear Information System (INIS)

Wagner, G.J.

1979-01-01

The mechanisms underlying heavy metal accumulation, toxicity, and tolerance in higher plants are poorly understood. Since subcellular processes are undoubtedly involved in all these phenomena, it is of interest to study the extent, subcellular site and nature of intracellularly accumulated cadmium in higher plants. Whole plants supplied 109 CdCl 2 or 112 CdSO 4 accumulated Cd into roots and aerial tissues. Preparation of protoplasts from aerial tissues followed by subcellular fractionation of the protoplasts to obtain intact vacuoles, chloroplasts and cytosol revealed the presence of Cd in the cytosol but not in vacuoles or chloroplasts. No evidence was obtained for the production of volatile Cd complexes in tobacco

COPPER RESISTANT STRAIN CANDIDA TROPICALIS RomCu5 INTERACTION WITH SOLUBLE AND INSOLUBLE COPPER COMPOUNDS

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Ie. P. Prekrasna

2015-10-01

Full Text Available The focus of the study was interaction of Candida tropicalis RomCu5 isolated from highland Ecuador ecosystem with soluble and insoluble copper compounds. Strain C. tropicalis RomCu5 was cultured in a liquid medium of Hiss in the presence of soluble (copper citrate and CuCl2 and insoluble (CuO and CuCO3 copper compounds. The biomass growth was determined by change in optical density of culture liquid, composition of the gas phase was measured on gas chromatograph, redox potential and pH of the culture fluid was defined potentiometrically. The concentration of soluble copper compounds was determined colorimetrically. Maximal permissible concentration of Cu2+ for C. tropicalis RomCu5 was 30 000 ppm of Cu2+ in form of copper citrate and 500 ppm of Cu2+ in form of CuCl2. C. tropicalis was metabolically active at super high concentrations of Cu2+, despite the inhibitory effect of Cu2+. C. tropicalis immobilized Cu2+ in the form of copper citrate and CuCl2 by it accumulation in the biomass. Due to medium acidification C. tropicalis dissolved CuO and CuCO3. High resistance of C. tropicalis to Cu2+ and ability to interact with soluble and insoluble copper compounds makes it biotechnologically perspective.

Influence of copper high-tension lines on plants and soils

Energy Technology Data Exchange (ETDEWEB)

Kraal, H.; Ernst, W.

1976-09-01

The copper contents of plants and soils were determined in relation to the distance from copper high-tension lines. In the vicinity of the cables clayey and fenny soils had demonstrably higher copper contents, due to corrosion of the cables, than regions 20 m and more outside the high-tension lines. On these soils, however, copper accumulation in the plants was low in comparison with those from a sandy soil, although this soil itself showed no copper increase in relation to the cables. The contaminated plants may present a risk of poisoning for sheep within a 20 m distance on both sides of the cables. No changes in plant species composition and in the copper tolerance of Agrostis tenuis were observed.

Biochemical diagnosis of mucopolysaccharidoses by estimation of intracellular 35S-sulfate accumulation

International Nuclear Information System (INIS)

Petruschka, L.; Machill, G.; Wehnert, M.

1985-01-01

The investigation of 35 S-sulfate accumulation and of 35 SO 4 -labelled glycosaminoglucane molecules (chase-experiments) in cultured cells was used in post- and prenatal diagnosis of mucopolysaccharidosis (MPS). Cell lines which accumulate 35 S-sulfate can be differentiated by means of cross correction or measurement of enzyme activity. 34 patients with the MPS type I, II, III A, III B and VI, respectively, were diagnosed in this way. Four pregnancies at risk for MPS were prenatally monitored by the sulfate accumulation in cultured amniotic fluid cells. One fetus suffering from MPS II was found. (author)

Microbial accumulation of uranium, radium, and cesium

International Nuclear Information System (INIS)

Strandberg, G.W.; Shumate, S.E. II; Parrott, J.R. Jr.; North, S.E.

1981-05-01

Diverse microbial species varied considerably in their ability to accumulate uranium, cesium, and radium. Mechanistic differences in uranium uptake by Saccharomyces cerevisiae and Pseudomonas aeruginosa were indicated. S. serevisiae exhibited a slow (hours) surface accumulation of uranium which was subject to environmental factors, while P. aeruginosa accumulated uranium rapidly (minutes) as dense intracellular deposits and did not appear to be affected by environmental parameters. Metabolism was not required for uranium uptake by either organism. Cesium and radium were concentrated to a considerably lesser extent than uranium by the several species tested

Accumulation of heavy metals (cadmium, zinc, and copper) from smelter in forest ecosystems and their uptakes by Shiitake mushroom (Lentinus edodes (Berk) Sing. ) and Nameko mushroom (Pholiota glutinosa Kawamura) through polluted bed logs

Energy Technology Data Exchange (ETDEWEB)

Nishimoto, T.; Fujita, K.; Furukawa, H.; Yoshimoto, M.

1977-12-01

Mushrooms cultivated on sawdust medium which had been innoculated with heavy metals accumulated the metals increasingly in stems, pileus, gill and spores, in that order. There were strain differences, in accumulation, and highest concentration was found in the first-born fruit body. At 2 ppm, cadmium did not affect yield of the fruiting body. At 20 ppm, however, yield was seriously reduced. Species differences in absorption capacity for heavy metals were noted. Seasonal variations in cadmium and copper accumulation were noted, along with zinc. Cadmium concentration in fruiting bodies increased with increase of cadmium concentration in the growth substrate. 23 figures, 16 tables.

Copper(II)-bis(thiosemicarbazonato) complexes as anti-chlamydial agents.

Science.gov (United States)

Marsh, James W; Djoko, Karrera Y; McEwan, Alastair G; Huston, Wilhelmina M

2017-09-29

Lipophilic copper (Cu)-containing complexes have shown promising antibacterial activity against a range of bacterial pathogens. To examine the susceptibility of the intracellular human pathogen Chlamydia trachomatis to copper complexes containing bis(thiosemicarbazone) ligands [Cu(btsc)], we tested the in vitro effect of CuII-diacetyl- and CuII-glyoxal-bis[N(4)-methylthiosemicarbazonato] (Cu(atsm) and Cu(gtsm), respectively) on C. trachomatis. Cu(atsm) and to a greater extent, Cu(gtsm), prevented the formation of infectious chlamydial progeny. Impacts on host cell viability and respiration were also observed in addition to the Chlamydia impacts. This work suggests that copper-based complexes may represent a new lead approach for future development of new therapeutics against chlamydial infections, although host cell impacts need to be fully explored. © FEMS 2017. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

Stabilized copper plating method by programmed electroplated current: Accumulation of densely packed copper grains in the interconnect

Energy Technology Data Exchange (ETDEWEB)

Kao, Li-Chi; Hsu, Li-Hsuan; Brahma, Sanjaya; Huang, Bo-Chia; Liu, Chun-Chu; Lo, Kuang-Yao, E-mail: kuanglo@mail.ncku.edu.tw

2016-12-01

Highlights: • Actual Cu interconnect experiences many times of annealing and then cause the stress. • Stack Cu grains with varying grain size successively to enhance packed density. • XRD and PBR analyze the residual stress of local and average area of plated Cu film. • High packed Cu grain with stable stress proved by texture of Cu(1 1 1) and Cu(2 0 0). - Abstract: In this work, we programmed the plating current to stack the different size of copper (Cu) grain and analyzed the relation between the sequence of different Cu grain size and the stability of the residual stress. The residual stress was measured with varying times of annealing process in order to reach the purpose of simulating the actual Cu interconnect process. We found that varied plating strategy will make different stabilization condition of residual stress through the proof of X-ray diffraction (XRD) and optical parallel beams reflection (PBR) method. The accumulation of Cu grains, formed by Cu grain with successive variation in grain size, would enhance the packing density better than only single grain size in the finite space. The high density of the grain boundary in the electroplated Cu film will be eliminated through annealing process and it will help to suppress the void formation in further interconnect process. The electroplated Cu film with the plating current of saw tooth wave can soon reach a stable tensile stress through annealing since the Cu grains with high packing density will be quickly eliminated to approach the minimum of the strain energy which reflects to variation in the texture of Cu (2 0 0). The result of this work illustrates the importance of how to stack different size of Cu grain, for achieving a densely packed Cu film which close to the Cu bulk.

Heavy metal accumulation by three species of mosses in Shillong, Northeastern India

Energy Technology Data Exchange (ETDEWEB)

Gupta, A. [St. Edmund`s College, Shillong (India). Dept. of Zoology

1995-06-01

Comparisons were made of the accumulation of cadmium, copper, manganese, lead, and zinc in Plagiothecium denticulatum, Bryum argenteum and Sphagnum sp. in Shillong, Meghalaya State, Northeastern India. Samples of P. denticulatum and B. argenteum were collected inside Shillong city (urban) and its immediately adjacent outskirts (suburban), while Sphagnum sp. was collected from a suburban site only. Lead and copper levels were higher in P. denticulatum, while Sphagnum sp. accumulated higher amounts of zinc, manganese, and cadmium. An urban-suburban gradient was evident for lead and zinc in P. denticulatum, and for cadmium in B. argenteum, while a reverse trend could be discerned for manganese in P. denticulatum. Besides vehicles and minor industries, quarry dust was likely to be an important source of manganese, copper and zinc. 13 refs., 1 fig., 1 tab.

Adsorptive stripping voltammetric behaviour of copper complexes of some heterocyclic azo compounds.

Science.gov (United States)

Farias, P A; Ferreira, S L; Ohara, A K; Bastos, M B; Goulart, M S

1992-10-01

Controlled adsorptive accumulation of copper complexed with TAN, TAC, TAR and TAM (heterocyclic azo-compounds) on a static mercury drop electrode provides the basis for the direct stripping measurement of this element in the nanomolar concentration level. The ligand TAN exhibited great sensitivity and better separation of the peak current of the ligand in relation to the complex. The reduction current of adsorbed complex ions of copper is measured by linear scan cathodic stripping voltammetry, preceded by a period of accumulation of a few minutes. The peak potential is at approximately -0.37 V vs. Ag/AgCl. Optimal experimental parameters were found to be a TAN concentration of 1 x 10(-5)M, an accumulation potential of -0.22 V, and a solution pH of 3.7 (acetate buffer). The detection limit is 0.8nM after a 5-min accumulation with a stirred solution, and the response is linear up to 50 mug/l. Many common cations and anions do not interfere in the determination of copper. The interference of titanium is eliminated by addition of fluoride ion. Results are reported for a fresh water sample.

Urea increased nickel and copper accumulation in the leaves of Egeria densa (Planch.) Casp. and Ceratophyllum demersum L. during short-term exposure.

Science.gov (United States)

Maleva, Maria; Borisova, Galina; Chukina, Nadezhda; Kumar, Adarsh

2018-02-01

In the present study, two fresh water plant species Egeria densa (Planch.) Casp. and Ceratophyllum demersum L. were subjected to separate and combined action of urea (2mМ) and metals (Ni and Cu, 10μM) to investigate the phytoremediation potential of these two submerged macrophytes during short-term experiments (48h). Both submerged macrophytes demonstrated high accumulative potential for Ni and Cu (average bioconcentration factors were 2505 for Ni and 3778 for Cu). The urea (2 mM) was not significantly toxic for studied plant species. Futhermore, urea worked as an additional source of nitrogen and stimulated some metabolic processes such as the synthesis of photosynthetic pigments, soluble proteins, non-enzymatic antioxidants, and activated some enzymes. Adding urea to the metals increased their accumulation in both macrophytes (on average by 35% for Ni and 15% for Cu). Combined action of urea and Ni did not have a significant effect on antioxidant response, but caused a sharp increase of urease activity (4 folds on an average) in both plants. The copper exerted a stronger toxic effect on both studied macrophytes compared to nickel. Adding urea to copper in some cases diminished the toxic action of this metal. Study concludes that the responses of E. densa and C. demersum to urea and metal action (separate and combined) were depended on the type of pollutant and the activity of antioxidant defence system. Therefore, the studied aquatic macrophytes found to be potential phytoremediators of water bodies, the addition of an organic nitrogen source in the form of urea in environmentally relevant concentration will increase the efficiency of phytoextraction of metals. Copyright © 2017 Elsevier Inc. All rights reserved.

Copper induced oxidative stresses, antioxidant responses and phytoremediation potential of Moso bamboo (Phyllostachys pubescens)

Science.gov (United States)

Chen, Junren; Shafi, Mohammad; Li, Song; Wang, Ying; Wu, Jiasen; Ye, Zhengqian; Peng, Danli; Yan, Wenbo; Liu, Dan

2015-09-01

Moso bamboo is recognized as phytoremediation plant due to production of huge biomass and high tolerance in stressed environment. Hydroponics and pot experiments were conducted to investigate mechanism of copper tolerance and to evaluate copper accumulation capacity of Moso bamboo. In hydroponics experiment there was non significant variation in MDA contents of leaves compared with control. SOD and POD initially indicated enhancing trend with application of 5 μM Cu and then decreased consistently with application of 25 and 100 μM Cu. Application of each additional increment of copper have constantly enhanced proline contents while maximum increase of proline was observed with application of 100 μM copper. In pot experiment chlorophyll and biomass initially showed increasing tendency and decreased gradually with application of each additional increment of Cu. Normal growth of Moso bamboo was observed with application of 100 mg kg-1 copper. However, additional application of 300 or 600 mg kg-1 copper had significantly inhibited growth of Moso bamboo. The concentration of Cu in Moso bamboo has attained the levels of 340, 60, 23 mg kg-1 in roots, stems and leaves respectively. The vacuoles were the main organs which accumulated copper and reduced toxicity of copper as studied by TEM-DEX technology.

Cupriferous peat: embryonic copper ore

Energy Technology Data Exchange (ETDEWEB)

Fraser, D C

1961-07-01

A Canadian peat was found to contain up to 10% (dry weight) Cu, and a mechanism for Cu accumulation in peat was discussed. Wet chemical techniques and x-ray diffraction were utilized to identify Cu compounds. Copper was organically bound in peat as a chelate complex and did not occur as an oxide, sulfide, or as elemental Cu. Because of the low S content of peat the Cu was assumed to be bound to nitrogen or oxygen-containing components. Copper, having a greater affinity for N, tended to form the more stable Cu-N chelate. The element was concentrated as circulating cupriferous ground waters filtered through the peat.

Albumin has no role in the uptake of copper by human fibroblasts

International Nuclear Information System (INIS)

McArdle, H.J.; Guthrie, J.R.; Ackland, M.L.; Danks, D.M.

1987-01-01

The mechanism of copper uptake by cells has been the subject of controversy for some time. This paper examines the possibility of a role for albumin in the uptake of copper by fibroblasts. Although the cells could accumulate copper from a copper-albumin complex, there was no evidence for either copper-albumin or albumin receptors on the cell surface. The possibility of a surface exchange mechanism for copper was examined. While copper uptake showed saturation with increasing concentrations of labelled copper-albumin, adding unlabelled copper to the incubation medium did not inhibit uptake. Adding albumin or histidine to the copper-albumin complex resulted in an inhibition of copper uptake. The results can only be explained by the cell taking up free copper from the incubation medium, with the albumin then releasing its copper to maintain the equilibrium between free and bound metal. Since, in vivo there is essentially no free copper in serum, it is concluded that albumin is most unlikely to play a role in the uptake of copper by fibroblasts

Effect of copper on growth of an aquatic macrophyte, Elodea canadensis

Energy Technology Data Exchange (ETDEWEB)

Mal, T.K.; Adorjan, Peter; Corbett, A.L

2002-12-01

Elodea canadensis may be a good biomonitor for copper, but not a good bioaccumulator. - Elodea canadensis has been proposed as a potential biomonitor due to its wide distribution and apparent ability to accumulate pollutants in aquatic ecosystems. We investigated the effects of copper sulfate on growth in E. canadensis to determine its effectiveness as a biomonitor of copper pollution in aquatic systems and whether growth is a suitable index of sub-lethal stress. Copper sulfate significantly slowed or stopped growth at all concentrations (low: 1 ppm, medium: 5 ppm, high: 10 ppm of copper sulfate) used. Final plant drymass was significantly lower in medium and high copper treatments compared with controls. E. canadensis appears to be very sensitive to copper levels, and may be useful as a biomonitor of copper levels in aquatic systems. However, its utility as a bioaccumulator may be limited, because we observed senescence of most leaves in all copper-treated plants following 4 weeks of treatment.

Effect of copper on growth of an aquatic macrophyte, Elodea canadensis

International Nuclear Information System (INIS)

Mal, T.K.; Adorjan, Peter; Corbett, A.L.

2002-01-01

Elodea canadensis may be a good biomonitor for copper, but not a good bioaccumulator. - Elodea canadensis has been proposed as a potential biomonitor due to its wide distribution and apparent ability to accumulate pollutants in aquatic ecosystems. We investigated the effects of copper sulfate on growth in E. canadensis to determine its effectiveness as a biomonitor of copper pollution in aquatic systems and whether growth is a suitable index of sub-lethal stress. Copper sulfate significantly slowed or stopped growth at all concentrations (low: 1 ppm, medium: 5 ppm, high: 10 ppm of copper sulfate) used. Final plant drymass was significantly lower in medium and high copper treatments compared with controls. E. canadensis appears to be very sensitive to copper levels, and may be useful as a biomonitor of copper levels in aquatic systems. However, its utility as a bioaccumulator may be limited, because we observed senescence of most leaves in all copper-treated plants following 4 weeks of treatment

Effect of copper on growth of an aquatic macrophyte, Elodea canadensis.

Science.gov (United States)

Mal, Tarun K; Adorjan, Peter; Corbett, Andrea L

2002-01-01

Elodea canadensis has been proposed as a potential biomonitor due to its wide distribution and apparent ability to accumulate pollutants in aquatic ecosystems. We investigated the effects of copper sulfate on growth in E. canadensis to determine its effectiveness as a biomonitor of copper pollution in aquatic systems and whether growth is a suitable index of sub-lethal stress. Copper sulfate significantly slowed or stopped growth at all concentrations (low: 1 ppm, medium: 5 ppm, high: 10 ppm of copper sulfate) used. Final plant drymass was significantly lower in medium and high copper treatments compared with controls. E. canadensis appears to be very sensitive to copper levels, and may be useful as a biomonitor of copper levels in aquatic systems. However, its utility as a bioaccumulator may be limited, because we observed senescence of most leaves in all copper-treated plants following 4 weeks of treatment.

CopM is a novel copper-binding protein involved in copper resistance in Synechocystis sp. PCC 6803

Science.gov (United States)

Giner-Lamia, Joaquín; López-Maury, Luis; Florencio, Francisco J

2015-01-01

Copper resistance system in the cyanobacterium Synechocystis sp. PCC 6803 comprises two operons, copMRS and copBAC, which are expressed in response to copper in the media. copBAC codes for a heavy-metal efflux–resistance nodulation and division (HME-RND) system, while copMRS codes for a protein of unknown function, CopM, and a two-component system CopRS, which controls the expression of these two operons. Here, we report that CopM is a periplasmic protein able to bind Cu(I) with high affinity (KD ∼3 × 10−16). Mutants lacking copM showed a sensitive copper phenotype similar to mutants affected in copB, but lower than mutants of the two-component system CopRS, suggesting that CopBAC and CopM constitute two independent resistance mechanisms. Moreover, constitutive expression of copM is able to partially suppress the copper sensitivity of the copR mutant strain, pointing out that CopM per se is able to confer copper resistance. Furthermore, constitutive expression of copM was able to reduce total cellular copper content of the copR mutant to the levels determined in the wild-type (WT) strain. Finally, CopM was localized not only in the periplasm but also in the extracellular space, suggesting that CopM can also prevent copper accumulation probably by direct copper binding outside the cell. PMID:25545960

Primary biochemical defect in copper metabolism in mice with a recessive X-linked mutation analogous to Menkes' disease in man

International Nuclear Information System (INIS)

Prins, H.W.; Hamer, C.J.A. van den.

1979-01-01

The defect in Menkes' disease in man is identical to that in Brindled mice. The defect manifests itself in a accumulation of copper in some tissues, such as renal, intestinal (mucosa and muscle), pancreatic, osseous, muscular, and dermal. Hence a fatal copper deficiency results in other tissues (e.g., hepatic). The copper transport through the intestine is impaired and copper, which circumvents the block in the copper resorption, is irreversibly trapped in the above-mentioned, copper accumulating tissues where it is bound to a cytoplasmatic protein with molecular weight 10,000 daltons, probably the primary cytoplasmatic copper transporting protein. This protein shows a Cu-S absorption band at 250 nm, and the copper:protein ratio is increased. Such copper rich protein was found neither in the kidneys of the unaffected mica nor in the liver of the mice that do have the defect. Three models of the primary defect in Menkes' disease are proposed

Intracellular nitrate of marine diatoms as a driver of anaerobic nitrogen cycling in sinking aggregates

DEFF Research Database (Denmark)

Kamp, Anja; Stief, Peter; Bristow, Laura A.

2016-01-01

% was recovered as nitrite. Hence, aggregate-associated diatoms accumulate nitrate from the surrounding water and sustain complex nitrogen transformations, including loss of fixed nitrogen, in anoxic, pelagic microniches. Additionally, it may be expected that intracellular nitrate not converted before...... store nitrate intracellularly, we explored the fate of intracellular nitrate and its availability for microbial metabolism within anoxic diatom-bacteria aggregates. The ubiquitous nitrate-storing diatom Skeletonema marinoi was studied as both axenic cultures and laboratory-produced diatom......-bacteria aggregates. Stable 15N isotope incubations under dark and anoxic conditions revealed that axenic S. marinoi is able to reduce intracellular nitrate to ammonium that is immediately excreted by the cells. When exposed to a light:dark cycle and oxic conditions, S. marinoi stored nitrate intracellularly...

Enhanced accumulation of copper and lead in amaranth (Amaranthus paniculatus), Indian mustard (Brassica juncea) and sunflower (Helianthus annuus).

Science.gov (United States)

Rahman, Motior M; Azirun, Sofian M; Boyce, Amru N

2013-01-01

Soil contamination by copper (Cu) and lead (Pb) is a widespread environmental problem. For phytoextraction to be successful and viable in environmental remediation, strategies that can improve plant uptake must be identified. In the present study we investigated the use of nitrogen (N) fertilizer as an efficient way to enhance accumulation of Cu and Pb from contaminated industrial soils into amaranth, Indian mustard and sunflower. Plants were grown in a greenhouse and fertilized with N fertilizer at rates of 0, 190 and 380 mg kg⁻¹ soil. Shoots, roots and total accumulation of Cu and Pb, transfer factor (TF), translocation index were assessed to evaluate the transport and translocation ability of tested plants. Addition of N fertilizer acidified the industrial soil and caused the pH to decrease to 5.5 from an initial pH of 6.9. Industrial soil amended with N fertilizer resulted in the highest accumulation of Pb and Cu (for Pb 10.1-15.5 mg kg⁻¹, for Cu 11.6-16.8 mg kg⁻¹) in the shoots, which was two to four folds higher relative to the concentration in roots in all the three plants used. Sunflower removed significantly higher Pb (50-54%) and Cu (34-38%) followed by amaranth and Indian mustard from industrial soils with the application of N fertilizer. The TF was Sunflower is the best plant species to carry out phytoextraction of Pb and Cu. In contrast, Pb and Cu removal by Indian mustard and amaranth shows great potential as quick and short duration vegetable crops. The results suggest that the application of N fertilizer in contaminated industrial soil is an effective amendment for the phytoextraction of Pb and Cu from contaminated industrial soils.

Combining -Omics to Unravel the Impact of Copper Nutrition on Alfalfa (Medicago sativa) Stem Metabolism.

Science.gov (United States)

Printz, Bruno; Guerriero, Gea; Sergeant, Kjell; Audinot, Jean-Nicolas; Guignard, Cédric; Renaut, Jenny; Lutts, Stanley; Hausman, Jean-Francois

2016-02-01

Copper can be found in the environment at concentrations ranging from a shortage up to the threshold of toxicity for plants, with optimal growth conditions situated in between. The plant stem plays a central role in transferring and distributing minerals, water and other solutes throughout the plant. In this study, alfalfa is exposed to different levels of copper availability, from deficiency to slight excess, and the impact on the metabolism of the stem is assessed by a non-targeted proteomics study and by the expression analysis of key genes controlling plant stem development. Under copper deficiency, the plant stem accumulates specific copper chaperones, the expression of genes involved in stem development is decreased and the concentrations of zinc and molybdenum are increased in comparison with the optimum copper level. At the optimal copper level, the expression of cell wall-related genes increases and proteins playing a role in cell wall deposition and in methionine metabolism accumulate, whereas copper excess imposes a reduction in the concentration of iron in the stem and a reduced abundance of ferritins. Secondary ion mass spectrometry (SIMS) analysis suggests a role for the apoplasm as a copper storage site in the case of copper toxicity. © The Author 2016. Published by Oxford University Press on behalf of Japanese Society of Plant Physiologists.

Copper induces hepatocyte injury due to the endoplasmic reticulum stress in cultured cells and patients with Wilson disease

International Nuclear Information System (INIS)

Oe, Shinji; Miyagawa, Koichiro; Honma, Yuichi; Harada, Masaru

2016-01-01

Copper is an essential trace element, however, excess copper is harmful to human health. Excess copper-derived oxidants contribute to the progression of Wilson disease, and oxidative stress induces accumulation of abnormal proteins. It is known that the endoplasmic reticulum (ER) plays an important role in proper protein folding, and that accumulation of misfolded proteins disturbs ER homeostasis resulting in ER stress. However, copper-induced ER homeostasis disturbance has not been fully clarified. We treated human hepatoma cell line (Huh7) and immortalized-human hepatocyte cell line (OUMS29) with copper and chemical chaperones, including 4-phenylbutyrate and ursodeoxycholic acid. We examined copper-induced oxidative stress, ER stress and apoptosis by immunofluorescence microscopy and immunoblot analyses. Furthermore, we examined the effects of copper on carcinogenesis. Excess copper induced not only oxidative stress but also ER stress. Furthermore, excess copper induced DNA damage and reduced cell proliferation. Chemical chaperones reduced this copper-induced hepatotoxicity. Excess copper induced hepatotoxicity via ER stress. We also confirmed the abnormality of ultra-structure of the ER of hepatocytes in patients with Wilson disease. These findings show that ER stress plays a pivotal role in Wilson disease, and suggests that chemical chaperones may have beneficial effects in the treatment of Wilson disease.

Copper induces hepatocyte injury due to the endoplasmic reticulum stress in cultured cells and patients with Wilson disease

Energy Technology Data Exchange (ETDEWEB)

Oe, Shinji, E-mail: ooes@med.uoeh-u.ac.jp; Miyagawa, Koichiro, E-mail: koichiro@med.uoeh-u.ac.jp; Honma, Yuichi, E-mail: y-homma@med.uoeh-u.ac.jp; Harada, Masaru, E-mail: msrharada@med.uoeh-u.ac.jp

2016-09-10

Copper is an essential trace element, however, excess copper is harmful to human health. Excess copper-derived oxidants contribute to the progression of Wilson disease, and oxidative stress induces accumulation of abnormal proteins. It is known that the endoplasmic reticulum (ER) plays an important role in proper protein folding, and that accumulation of misfolded proteins disturbs ER homeostasis resulting in ER stress. However, copper-induced ER homeostasis disturbance has not been fully clarified. We treated human hepatoma cell line (Huh7) and immortalized-human hepatocyte cell line (OUMS29) with copper and chemical chaperones, including 4-phenylbutyrate and ursodeoxycholic acid. We examined copper-induced oxidative stress, ER stress and apoptosis by immunofluorescence microscopy and immunoblot analyses. Furthermore, we examined the effects of copper on carcinogenesis. Excess copper induced not only oxidative stress but also ER stress. Furthermore, excess copper induced DNA damage and reduced cell proliferation. Chemical chaperones reduced this copper-induced hepatotoxicity. Excess copper induced hepatotoxicity via ER stress. We also confirmed the abnormality of ultra-structure of the ER of hepatocytes in patients with Wilson disease. These findings show that ER stress plays a pivotal role in Wilson disease, and suggests that chemical chaperones may have beneficial effects in the treatment of Wilson disease.

Copper/MYC/CTR1 interplay: a dangerous relationship in hepatocellular carcinoma.

Science.gov (United States)

Porcu, Cristiana; Antonucci, Laura; Barbaro, Barbara; Illi, Barbara; Nasi, Sergio; Martini, Maurizio; Licata, Anna; Miele, Luca; Grieco, Antonio; Balsano, Clara

2018-02-06

Free serum copper correlates with tumor incidence and progression of human cancers, including hepatocellular carcinoma (HCC). Copper extracellular uptake is provided by the transporter CTR1, whose expression is regulated to avoid excessive intracellular copper entry. Inadequate copper serum concentration is involved in the pathogenesis of Non Alcoholic Fatty Liver Disease (NAFLD), which is becoming a major cause of liver damage progression and HCC incidence. Finally, MYC is over-expressed in most of HCCs and is a critical regulator of cellular growth, tumor invasion and metastasis. The purpose of our study was to understand if higher serum copper concentrations might be involved in the progression of NAFLD-cirrhosis toward-HCC. We investigated whether high exogenous copper levels sensitize liver cells to transformation and if it exists an interplay between copper-related proteins and MYC oncogene. NAFLD-cirrhotic patients were characterized by a statistical significant enhancement of serum copper levels, even more evident in HCC patients. We demonstrated that high extracellular copper concentrations increase cell growth, migration, and invasion of liver cancer cells by modulating MYC/CTR1 axis. We highlighted that MYC binds a specific region of the CTR1 promoter, regulating its transcription. Accordingly, CTR1 and MYC proteins expression were progressively up-regulated in liver tissues from NAFLD-cirrhotic to HCC patients. This work provides novel insights on the molecular mechanisms by which copper may favor the progression from cirrhosis to cancer. The Cu/MYC/CTR1 interplay opens a window to refine HCC diagnosis and design new combined therapies.

Copper/MYC/CTR1 interplay: a dangerous relationship in hepatocellular carcinoma

Science.gov (United States)

Barbaro, Barbara; Illi, Barbara; Nasi, Sergio; Martini, Maurizio; Licata, Anna; Miele, Luca; Grieco, Antonio; Balsano, Clara

2018-01-01

Free serum copper correlates with tumor incidence and progression of human cancers, including hepatocellular carcinoma (HCC). Copper extracellular uptake is provided by the transporter CTR1, whose expression is regulated to avoid excessive intracellular copper entry. Inadequate copper serum concentration is involved in the pathogenesis of Non Alcoholic Fatty Liver Disease (NAFLD), which is becoming a major cause of liver damage progression and HCC incidence. Finally, MYC is over-expressed in most of HCCs and is a critical regulator of cellular growth, tumor invasion and metastasis. The purpose of our study was to understand if higher serum copper concentrations might be involved in the progression of NAFLD-cirrhosis toward-HCC. We investigated whether high exogenous copper levels sensitize liver cells to transformation and if it exists an interplay between copper-related proteins and MYC oncogene. NAFLD-cirrhotic patients were characterized by a statistical significant enhancement of serum copper levels, even more evident in HCC patients. We demonstrated that high extracellular copper concentrations increase cell growth, migration, and invasion of liver cancer cells by modulating MYC/CTR1 axis. We highlighted that MYC binds a specific region of the CTR1 promoter, regulating its transcription. Accordingly, CTR1 and MYC proteins expression were progressively up-regulated in liver tissues from NAFLD-cirrhotic to HCC patients. This work provides novel insights on the molecular mechanisms by which copper may favor the progression from cirrhosis to cancer. The Cu/MYC/CTR1 interplay opens a window to refine HCC diagnosis and design new combined therapies. PMID:29507693

The copYAZ Operon Functions in Copper Efflux, Biofilm Formation, Genetic Transformation, and Stress Tolerance in Streptococcus mutans

Science.gov (United States)

Singh, Kamna; Senadheera, Dilani B.; Lévesque, Céline M.

2015-01-01

ABSTRACT In bacteria, copper homeostasis is closely monitored to ensure proper cellular functions while avoiding cell damage. Most Gram-positive bacteria utilize the copYABZ operon for copper homeostasis, where copA and copB encode copper-transporting P-type ATPases, whereas copY and copZ regulate the expression of the cop operon. Streptococcus mutans is a biofilm-forming oral pathogen that harbors a putative copper-transporting copYAZ operon. Here, we characterized the role of copYAZ operon in the physiology of S. mutans and delineated the mechanisms of copper-induced toxicity in this bacterium. We observed that copper induced toxicity in S. mutans cells by generating oxidative stress and disrupting their membrane potential. Deletion of the copYAZ operon in S. mutans strain UA159 resulted in reduced cell viability under copper, acid, and oxidative stress relative to the viability of the wild type under these conditions. Furthermore, the ability of S. mutans to form biofilms and develop genetic competence was impaired under copper stress. Briefly, copper stress significantly reduced cell adherence and total biofilm biomass, concomitantly repressing the transcription of the gtfB, gtfC, gtfD, gbpB, and gbpC genes, whose products have roles in maintaining the structural and/or functional integrity of the S. mutans biofilm. Furthermore, supplementation with copper or loss of copYAZ resulted in significant reductions in transformability and in the transcription of competence-associated genes. Copper transport assays revealed that the ΔcopYAZ strain accrued significantly large amounts of intracellular copper compared with the amount of copper accumulation in the wild-type strain, thereby demonstrating a role for CopYAZ in the copper efflux of S. mutans. The complementation of the CopYAZ system restored copper expulsion, membrane potential, and stress tolerance in the copYAZ-null mutant. Taking these results collectively, we have established the function of the S. mutans

The copYAZ Operon Functions in Copper Efflux, Biofilm Formation, Genetic Transformation, and Stress Tolerance in Streptococcus mutans.

Science.gov (United States)

Singh, Kamna; Senadheera, Dilani B; Lévesque, Céline M; Cvitkovitch, Dennis G

2015-08-01

In bacteria, copper homeostasis is closely monitored to ensure proper cellular functions while avoiding cell damage. Most Gram-positive bacteria utilize the copYABZ operon for copper homeostasis, where copA and copB encode copper-transporting P-type ATPases, whereas copY and copZ regulate the expression of the cop operon. Streptococcus mutans is a biofilm-forming oral pathogen that harbors a putative copper-transporting copYAZ operon. Here, we characterized the role of copYAZ operon in the physiology of S. mutans and delineated the mechanisms of copper-induced toxicity in this bacterium. We observed that copper induced toxicity in S. mutans cells by generating oxidative stress and disrupting their membrane potential. Deletion of the copYAZ operon in S. mutans strain UA159 resulted in reduced cell viability under copper, acid, and oxidative stress relative to the viability of the wild type under these conditions. Furthermore, the ability of S. mutans to form biofilms and develop genetic competence was impaired under copper stress. Briefly, copper stress significantly reduced cell adherence and total biofilm biomass, concomitantly repressing the transcription of the gtfB, gtfC, gtfD, gbpB, and gbpC genes, whose products have roles in maintaining the structural and/or functional integrity of the S. mutans biofilm. Furthermore, supplementation with copper or loss of copYAZ resulted in significant reductions in transformability and in the transcription of competence-associated genes. Copper transport assays revealed that the ΔcopYAZ strain accrued significantly large amounts of intracellular copper compared with the amount of copper accumulation in the wild-type strain, thereby demonstrating a role for CopYAZ in the copper efflux of S. mutans. The complementation of the CopYAZ system restored copper expulsion, membrane potential, and stress tolerance in the copYAZ-null mutant. Taking these results collectively, we have established the function of the S. mutans Cop

Metal accumulation in the earthworm Lumbricus rubellus. Model predictions compared to field data

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Veltman, K.; Huijbregts, M.A.J.; Vijver, M.G.; Peijnenburg, W.J.G.M.; Hobbelen, P.H.F.; Koolhaas, J.E.; van Gestel, C.A.M.; van Vliet, P.C.J.; Jan, Hendriks A.

2007-01-01

The mechanistic bioaccumulation model OMEGA (Optimal Modeling for Ecotoxicological Applications) is used to estimate accumulation of zinc (Zn), copper (Cu), cadmium (Cd) and lead (Pb) in the earthworm Lumbricus rubellus. Our validation to field accumulation data shows that the model accurately predicts internal cadmium concentrations. In addition, our results show that internal metal concentrations in the earthworm are less than linearly (slope < 1) related to the total concentration in soil, while risk assessment procedures often assume the biota-soil accumulation factor (BSAF) to be constant. Although predicted internal concentrations of all metals are generally within a factor 5 compared to field data, incorporation of regulation in the model is necessary to improve predictability of the essential metals such as zinc and copper. ?? 2006 Elsevier Ltd. All rights reserved.

Intracellular vesicles as reproduction elements in cell wall-deficient L-form bacteria.

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Yves Briers

Full Text Available Cell wall-deficient bacteria, or L-forms, represent an extreme example of bacterial plasticity. Stable L-forms can multiply and propagate indefinitely in the absence of a cell wall. Data presented here are consistent with the model that intracellular vesicles in Listeria monocytogenes L-form cells represent the actual viable reproductive elements. First, small intracellular vesicles are formed along the mother cell cytoplasmic membrane, originating from local phospholipid accumulation. During growth, daughter vesicles incorporate a small volume of the cellular cytoplasm, and accumulate within volume-expanding mother cells. Confocal Raman microspectroscopy demonstrated the presence of nucleic acids and proteins in all intracellular vesicles, but only a fraction of which reveals metabolic activity. Following collapse of the mother cell and release of the daughter vesicles, they can establish their own membrane potential required for respiratory and metabolic processes. Premature depolarization of the surrounding membrane promotes activation of daughter cell metabolism prior to release. Based on genome resequencing of L-forms and comparison to the parental strain, we found no evidence for predisposing mutations that might be required for L-form transition. Further investigations revealed that propagation by intracellular budding not only occurs in Listeria species, but also in L-form cells generated from different Enterococcus species. From a more general viewpoint, this type of multiplication mechanism seems reminiscent of the physicochemical self-reproducing properties of abiotic lipid vesicles used to study the primordial reproduction pathways of putative prokaryotic precursor cells.

Intracellular vesicles as reproduction elements in cell wall-deficient L-form bacteria.

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Briers, Yves; Staubli, Titu; Schmid, Markus C; Wagner, Michael; Schuppler, Markus; Loessner, Martin J

2012-01-01

Cell wall-deficient bacteria, or L-forms, represent an extreme example of bacterial plasticity. Stable L-forms can multiply and propagate indefinitely in the absence of a cell wall. Data presented here are consistent with the model that intracellular vesicles in Listeria monocytogenes L-form cells represent the actual viable reproductive elements. First, small intracellular vesicles are formed along the mother cell cytoplasmic membrane, originating from local phospholipid accumulation. During growth, daughter vesicles incorporate a small volume of the cellular cytoplasm, and accumulate within volume-expanding mother cells. Confocal Raman microspectroscopy demonstrated the presence of nucleic acids and proteins in all intracellular vesicles, but only a fraction of which reveals metabolic activity. Following collapse of the mother cell and release of the daughter vesicles, they can establish their own membrane potential required for respiratory and metabolic processes. Premature depolarization of the surrounding membrane promotes activation of daughter cell metabolism prior to release. Based on genome resequencing of L-forms and comparison to the parental strain, we found no evidence for predisposing mutations that might be required for L-form transition. Further investigations revealed that propagation by intracellular budding not only occurs in Listeria species, but also in L-form cells generated from different Enterococcus species. From a more general viewpoint, this type of multiplication mechanism seems reminiscent of the physicochemical self-reproducing properties of abiotic lipid vesicles used to study the primordial reproduction pathways of putative prokaryotic precursor cells.

Micronized copper wood preservatives: An efficiency and potential health risk assessment for copper-based nanoparticles

International Nuclear Information System (INIS)

Civardi, Chiara; Schwarze, Francis W.M.R.; Wick, Peter

2015-01-01

Copper (Cu) is an essential biocide for wood protection, but fails to protect wood against Cu-tolerant wood-destroying fungi. Recently Cu particles (size range: 1 nm–25 μm) were introduced to the wood preservation market. The new generation of preservatives with Cu-based nanoparticles (Cu-based NPs) is reputedly more efficient against wood-destroying fungi than conventional formulations. Therefore, it has the potential to become one of the largest end uses for wood products worldwide. However, during decomposition of treated wood Cu-based NPs and/or their derivate may accumulate in the mycelium of Cu-tolerant fungi and end up in their spores that are dispersed into the environment. Inhaled Cu-loaded spores can cause harm and could become a potential risk for human health. We collected evidence and discuss the implications of the release of Cu-based NPs by wood-destroying fungi and highlight the exposure pathways and subsequent magnitude of health impact. - Highlights: • We compared copper particulate wood preservatives with conventional ones. • We assessed the fungicidal activity of particulate copper wood preservatives. • We reviewed the Cu-tolerance mechanisms of some wood-destroying fungi. • Fungi colonizing wood treated with particulate copper may release Cu-loaded spores. - We assess the fungicidal activity of particulate copper wood preservatives and their possible release in the air by Cu-tolerant wood-destroying fungi

Yeast CUP1 protects HeLa cells against copper-induced stress

Energy Technology Data Exchange (ETDEWEB)

Xie, X.X. [Department of Animal Sciences, School of Agriculture and Biology, Shanghai Jiao Tong University, Shanghai (China); Shanghai Key Laboratory of Veterinary Biotechnology, Shanghai (China); College of Biological and Environmental Engineering, Zhejiang University of Technology, Hangzhou (China); Ma, Y.F.; Wang, Q.S.; Chen, Z.L.; Liao, R.R.; Pan, Y.C. [Department of Animal Sciences, School of Agriculture and Biology, Shanghai Jiao Tong University, Shanghai (China); Shanghai Key Laboratory of Veterinary Biotechnology, Shanghai (China)

2015-06-12

As an essential trace element, copper can be toxic in mammalian cells when present in excess. Metallothioneins (MTs) are small, cysteine-rich proteins that avidly bind copper and thus play an important role in detoxification. YeastCUP1 is a member of the MT gene family. The aim of this study was to determine whether yeast CUP1 could bind copper effectively and protect cells against copper stress. In this study,CUP1 expression was determined by quantitative real-time PCR, and copper content was detected by inductively coupled plasma mass spectrometry. Production of intracellular reactive oxygen species (ROS) was evaluated using the 2',7'-dichlorofluorescein-diacetate (DCFH-DA) assay. Cellular viability was detected using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay, and the cell cycle distribution of CUP1 was analyzed by fluorescence-activated cell sorting. The data indicated that overexpression of yeast CUP1 in HeLa cells played a protective role against copper-induced stress, leading to increased cellular viability (P<0.05) and decreased ROS production (P<0.05). It was also observed that overexpression of yeast CUP1 reduced the percentage of G1 cells and increased the percentage of S cells, which suggested that it contributed to cell viability. We found that overexpression of yeast CUP1 protected HeLa cells against copper stress. These results offer useful data to elucidate the mechanism of the MT gene on copper metabolism in mammalian cells.

Model Linking Plasma and Intracellular Tenofovir/Emtricitabine with Deoxynucleoside Triphosphates.

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Xinhui Chen

Full Text Available The coformulation of the nucleos(tide analogs (NA tenofovir (TFV disoproxil fumarate (TDF and emtricitabine (FTC is approved for HIV-infection treatment and prevention. Plasma TFV and FTC undergo complicated hybrid processes to form, accumulate, and retain as their active intracellular anabolites: TFV-diphosphate (TFV-DP and FTC-triphosphate (FTC-TP. Such complexities manifest in nonlinear intracellular pharmacokinetics (PK. In target cells, TFV-DP/FTC-TP compete with endogenous deoxynucleoside triphosphates (dNTP at the active site of HIV reverse transcriptase, underscoring the importance of analog:dNTP ratios for antiviral efficacy. However, NA such as TFV and FTC have the potential to disturb the dNTP pool, which could augment or reduce their efficacies. We conducted a pharmacokinetics-pharmacodynamics (PKPD study among forty subjects receiving daily TDF/FTC (300 mg/200 mg from the first-dose to pharmacological intracellular steady-state (30 days. TFV/FTC in plasma, TFV-DP/FTC-TP and dNTPs in peripheral blood mononuclear cells (PBMC were quantified using validated LC/MS/MS methodologies. Concentration-time data were analyzed using nonlinear mixed effects modeling (NONMEM. Formations and the accumulation of intracellular TFV-DP/FTC-TP was driven by plasma TFV/FTC, which was described by a hybrid of first-order formation and saturation. An indirect response link model described the interplay between TFV-DP/FTC-TP and the dNTP pool change. The EC50 (interindividual variability, (%CV of TFV-DP and FTC-TP on the inhibition of deoxyadenosine triphosphate (dATP and deoxycytidine triphosphate (dCTP production were 1020 fmol/106 cells (130% and 44.4 pmol/106 cells (82.5%, resulting in (90% prediction interval 11% (0.45%, 53% and 14% (2.6%, 35% reductions. Model simulations of analog:dNTP molar ratios using IPERGAY dosing suggested that FTC significantly contributes to the protective effect of preexposure prophylaxis (PrEP. Simulation

Temporal aspects of copper homeostasis and its crosstalk with hormones

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Lola ePeñarrubia

2015-04-01

Full Text Available To cope with the dual nature of copper as being essential and toxic for cells, plants temporarily adapt the expression of copper homeostasis components to assure its delivery to cuproproteins while avoiding the interference of potential oxidative damage derived from both copper uptake and photosynthetic reactions during light hours. The circadian clock participates in the temporal organization of coordination of plant nutrition adapting metabolic responses to the daily oscillations. This timely control improves plant fitness and reproduction and holds biotechnological potential to drive increased crop yields. Hormonal pathways, including those of abscisic acid, gibberellins, ethylene, auxins, and jasmonates are also under direct clock and light control, both in mono and dicotyledons. In this review, we focus on copper transport in Arabidopsis thaliana and Oryza sativa and the presumable role of hormones in metal homeostasis matching nutrient availability to growth requirements and preventing metal toxicity. The presence of putative hormone-dependent regulatory elements in the promoters of copper transporters genes suggests hormonal regulation to match special copper requirements during plant development. Spatial and temporal processes that can be affected by hormones include the regulation of copper uptake into roots, intracellular trafficking and compartmentalisation, and long-distance transport to developing vegetative and reproductive tissues. In turn, hormone biosynthesis and signalling are also influenced by copper availability, which suggests reciprocal regulation subjected to temporal control by the central oscillator of the circadian clock. This transcriptional regulatory network, coordinates environmental and hormonal signalling with developmental pathways to allow enhanced micronutrient acquisition efficiency.

Prenatal treatment of mosaic mice (Atp7a mo-ms) mouse model for Menkes disease, with copper combined by dimethyldithiocarbamate (DMDTC)

DEFF Research Database (Denmark)

Lenartowicz, Małgorzata; Krzeptowski, Wojciech; Koteja, Paweł

2012-01-01

Menkes disease is a fatal neurodegenerative disorder in infants caused by mutations in the gene ATP7A which encodes a copper (Cu) transporter. Defects in ATP7A lead to accumulated copper in the small intestine and kidneys and to copper deficiencies in the brain and the liver. The copper level...... and 18 with a combination of CuCl(2) (50 mg/kg) and dimethyldithiocarbamate (DMDTC) (280 mg/kg) leads to an increase in survival to about 76±25.3 days, whereas treatment with CuCl(2) alone (50 mg/kg) only leads to survival for about 21 days ±5 days. These copper-DMDTC treated mutants showed an improved...... locomotor activity performance and a gain in body mass. In contrast to treatment with CuCl(2) alone, a significant increase in the amount of copper was observed in the brain after prenatal copper-DMDTC treatment as well as a decrease in the amount of accumulated copper in the kidney, both leading towards...

Soil Copper and Zinc Accumulation and Bioavailability under a Long Term Vineyard Cultivation in South Italy

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Paolo Lorenzoni

2007-03-01

Full Text Available Soil metal contamination, particularly by copper, is a phenomenon which often occurs in the surface layer of vineyard soils, due to the widespread application of Cu-based products in the plant disease management. Our study was focused on soil Cu and Zn accumulation and bioavailability as related to some soil properties under a long term vineyard cultivation, in a D.O.C. wine area of South Italy (Calabria region. Soils selected from different landscape units, ranging from acid to alkaline, under homogeneous climate conditions and vineyard management system, were investigated. Each soil was sampled in both a vineyard and a fallow area, at the depth levels of 0-10 cm, 10-25 cm and 25-50 cm. The experimental data were analysed by ANOVA, correlation and multiple stepwise regression procedures. As expected, the results indicated a contamination of the vineyard soils by Cu due to the repeated application of Cu-based products in the plant disease control, with increments of total Cu content up to 150% against the fallow soils. On the contrary, the results led to exclude any soil Zn pollution due to the vineyard management and to suppose a main pedogenic origin for this metal. According to the relationships between Cu content and soil properties, Cu accumulation was promoted by higher pH, clay and organic matter contents. These soil properties also showed a strong influence on metal bioavailability, which underwent a significant reduction in soils with higher pH and clay contents. A further result of great significance was the adverse impact of soil erosion, enhanced by the application of not suitable management systems in hilly areas, on soil capability to retain polluting metals. Soil pH, organic matter content and texture, as well as soil management system, are key factors in soil capability to limit polluting metal dispersion in the environment.

Soil Copper and Zinc Accumulation and Bioavailability under a Long Term Vineyard Cultivation in South Italy

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Anna Maria Corea

2011-02-01

Full Text Available Soil metal contamination, particularly by copper, is a phenomenon which often occurs in the surface layer of vineyard soils, due to the widespread application of Cu-based products in the plant disease management. Our study was focused on soil Cu and Zn accumulation and bioavailability as related to some soil properties under a long term vineyard cultivation, in a D.O.C. wine area of South Italy (Calabria region. Soils selected from different landscape units, ranging from acid to alkaline, under homogeneous climate conditions and vineyard management system, were investigated. Each soil was sampled in both a vineyard and a fallow area, at the depth levels of 0-10 cm, 10-25 cm and 25-50 cm. The experimental data were analysed by ANOVA, correlation and multiple stepwise regression procedures. As expected, the results indicated a contamination of the vineyard soils by Cu due to the repeated application of Cu-based products in the plant disease control, with increments of total Cu content up to 150% against the fallow soils. On the contrary, the results led to exclude any soil Zn pollution due to the vineyard management and to suppose a main pedogenic origin for this metal. According to the relationships between Cu content and soil properties, Cu accumulation was promoted by higher pH, clay and organic matter contents. These soil properties also showed a strong influence on metal bioavailability, which underwent a significant reduction in soils with higher pH and clay contents. A further result of great significance was the adverse impact of soil erosion, enhanced by the application of not suitable management systems in hilly areas, on soil capability to retain polluting metals. Soil pH, organic matter content and texture, as well as soil management system, are key factors in soil capability to limit polluting metal dispersion in the environment.

Is a high serum copper concentration a risk factor for implantation failure?

Science.gov (United States)

Matsubayashi, Hidehiko; Kitaya, Kotaro; Yamaguchi, Kohei; Nishiyama, Rie; Takaya, Yukiko; Ishikawa, Tomomoto

2017-08-10

Copper-containing contraceptive devices may deposit copper ions in the endometrium, resulting in implantation failure. The deposition of copper ions in many organs has been reported in patients with untreated Wilson's disease. Since these patients sometimes exhibit subfertility and/or early pregnancy loss, copper ions were also considered to accumulate in the uterine endometrium. Wilson's disease patients treated with zinc successfully delivered babies because zinc interfered with the absorption of copper from the gastrointestinal tract. These findings led to the hypothesis that infertile patients with high serum copper concentrations may have implantation failure due to the excess accumulation of copper ions. The relationship between implantation (pregnancy) rates and serum copper concentrations has not yet been examined. The Japanese government recently stated that actual copper intake was higher among Japanese than needed. Therefore, the aim of the present study was to investigate whether serum copper concentrations are related to the implantation (pregnancy) rates of human embryos in vivo. We included 269 patients (age copper, and zinc concentrations were measured 16 days after the first date of progesterone replacement. We compared 96 women who were pregnant without miscarriage at 10 weeks of gestation (group P) and 173 women who were not pregnant (group NP). No significant differences were observed in age or BMI between the groups. Copper concentrations were significantly higher in group NP (average 193.2 μg/dL) than in group P (average 178.1 μg/dL). According to the area under the curve (AUC) on the receiver operating characteristic curve for the prediction of clinical pregnancy rates, the Cu/Zn ratio (AUC 0.64, 95% CI 0.54-0.71) was a better predictor than copper or zinc. When we set the cut-off as 1.59/1.60 for the Cu/Zn ratio, sensitivity, specificity, the positive predictive value, and negative predictive value were 0.98, 0.29, 0.71, and 0

Monitoring losses of copper based wood preservatives in the Thames estuary

International Nuclear Information System (INIS)

Hingston, J.A.; Murphy, R.J.; Lester, J.N.

2006-01-01

Field trials were conducted at two sites in the Thames estuary to monitor losses of copper, chromium and arsenic from wood preservative treated timbers of varying sizes and treatment regimes. Results indicated that leaching tests conducted under standard laboratory conditions might overestimate losses compared to losses resulting from real environmental exposures. Amine copper treated wood was noted to leach higher levels of copper compared to chromated copper arsenate treated wood, and was therefore considered an inappropriate replacement biocide for fresh and marine construction purposes on this basis. Increases in copper concentrations in the outer sections of amine copper treated posts may have represented re-distribution of this component in this timber. No accumulation of metals was found in sediments surrounding field trial posts. - Wood preservative field trials in the UK indicate that standard laboratory tests overestimate losses compared to those resulting from real environmental exposures

Monitoring losses of copper based wood preservatives in the Thames estuary

Energy Technology Data Exchange (ETDEWEB)

Hingston, J.A. [Environmental Processes and Water Technology Research Group, Department of Environmental Science and Technology, Faculty of Life Sciences, Imperial College, London SW7 2AZ (United Kingdom)]. E-mail: james.hingston@psd.defra.gsi.gov.uk; Murphy, R.J. [Department of Biology, Imperial College, London SW7 2AZ (United Kingdom); Lester, J.N. [School of Water Sciences, Cranfield University, Cranfield, Bedfordshire, MK43 0AL (United Kingdom)

2006-09-15

Field trials were conducted at two sites in the Thames estuary to monitor losses of copper, chromium and arsenic from wood preservative treated timbers of varying sizes and treatment regimes. Results indicated that leaching tests conducted under standard laboratory conditions might overestimate losses compared to losses resulting from real environmental exposures. Amine copper treated wood was noted to leach higher levels of copper compared to chromated copper arsenate treated wood, and was therefore considered an inappropriate replacement biocide for fresh and marine construction purposes on this basis. Increases in copper concentrations in the outer sections of amine copper treated posts may have represented re-distribution of this component in this timber. No accumulation of metals was found in sediments surrounding field trial posts. - Wood preservative field trials in the UK indicate that standard laboratory tests overestimate losses compared to those resulting from real environmental exposures.

Analysis method of intracellular iron accumulated in phytoplankton

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Jorge Falcão Paredes

1977-01-01

Full Text Available A colorimetric method for the assessment of iron accumulated in the cells of phytoplankton utilizing potassium ferrocyanide was developed. Two experiences with Phaeodactylum tricornutum and Chlorella sp. were carried out: the first one, on uptake of the iron relatively to the age of the culture: the second one, on the iron uptake by the cells, relatively to different concentrations of the chelator EDTA.Foi desenvolvido um método para análise do ferro acumulado em células de fitoplâncton, usando ferrocianeto de potássio como agente complexante. Com esse objetivo montaram-se duas experiências: uma em que se mostra o acúmulo de ferro em relação à idade das células e uma segunda experiência onde a assimilação é plotada em função da concentração de EDTA no meio de cultura.

Visualizing Metal Content and Intracellular Distribution in Primary Hippocampal Neurons with Synchrotron X-Ray Fluorescence.

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Robert A Colvin

Full Text Available Increasing evidence suggests that metal dyshomeostasis plays an important role in human neurodegenerative diseases. Although distinctive metal distributions are described for mature hippocampus and cortex, much less is known about metal levels and intracellular distribution in individual hippocampal neuronal somata. To solve this problem, we conducted quantitative metal analyses utilizing synchrotron radiation X-Ray fluorescence on frozen hydrated primary cultured neurons derived from rat embryonic cortex (CTX and two regions of the hippocampus: dentate gyrus (DG and CA1. Comparing average metal contents showed that the most abundant metals were calcium, iron, and zinc, whereas metals such as copper and manganese were less than 10% of zinc. Average metal contents were generally similar when compared across neurons cultured from CTX, DG, and CA1, except for manganese that was larger in CA1. However, each metal showed a characteristic spatial distribution in individual neuronal somata. Zinc was uniformly distributed throughout the cytosol, with no evidence for the existence of previously identified zinc-enriched organelles, zincosomes. Calcium showed a peri-nuclear distribution consistent with accumulation in endoplasmic reticulum and/or mitochondria. Iron showed 2-3 distinct highly concentrated puncta only in peri-nuclear locations. Notwithstanding the small sample size, these analyses demonstrate that primary cultured neurons show characteristic metal signatures. The iron puncta probably represent iron-accumulating organelles, siderosomes. Thus, the metal distributions observed in mature brain structures are likely the result of both intrinsic neuronal factors that control cellular metal content and extrinsic factors related to the synaptic organization, function, and contacts formed and maintained in each region.

Copper uptake by Pteris melanocaulon Fée from a Copper-Gold mine in Surigao del Norte, Philippines.

Science.gov (United States)

De la Torre, Joseph Benjamin B; Claveria, Rene Juna R; Perez, Rubee Ellaine C; Perez, Teresita R; Doronila, Augustine I

2016-01-01

The ability of some plants to take up metal contaminants in the soil has been of increasing interest as an environmental approach to pollution clean-up. This study aimed to assess the ability of Pteris melanocaulon for copper(Cu) uptake by determining the Cu levels in the fern vis-à-vis surrounding soil and the location of Cu accumulation within its biomass. It also aimed to add information to existing literature as P. melanocaulon are found to be less documented compared to other fern metal accumulators, such as P. vittata. The P. melanocaulon found in the Suyoc Pit of a Copper-Gold mine in Placer, Surigao del Norte, Philippines exhibited a high Bioaccumulation Factor(BF) of 4.04 and a low Translocation Factor(TF) of 0.01, suggesting more Cu accumulation in the roots (4590.22 ± 385.66 µg g(-1) Cu). Noteworthy was the Cu concentration in the rhizome which was also high (3539.44 ± 1696.35 µg g(-1) Cu). SEM/EDX analyses of the Cu content in the roots indicated high elemental %Cu in the xylem (6.95%) than in the cortex (2.68%). The high Cu content in the roots and rhizomes and the localization of Cu in the xylem manifested a potential utilization of the fern as a metallophyte for rhizofiltration and phytostabilization.

Full-length cellular β-secretase has a trimeric subunit stoichiometry, and its sulfur-rich transmembrane interaction site modulates cytosolic copper compartmentalization.

Science.gov (United States)

Liebsch, Filip; Aurousseau, Mark R P; Bethge, Tobias; McGuire, Hugo; Scolari, Silvia; Herrmann, Andreas; Blunck, Rikard; Bowie, Derek; Multhaup, Gerd

2017-08-11

The β-secretase (BACE1) initiates processing of the amyloid precursor protein (APP) into Aβ peptides, which have been implicated as central players in the pathology of Alzheimer disease. BACE1 has been described as a copper-binding protein and its oligomeric state as being monomeric, dimeric, and/or multimeric, but the native cellular stoichiometry has remained elusive. Here, by using single-molecule fluorescence and in vitro cross-linking experiments with photo-activatable unnatural amino acids, we show that full-length BACE1, independently of its subcellular localization, exists as trimers in human cells. We found that trimerization requires the BACE1 transmembrane sequences (TMSs) and cytoplasmic domains, with residues Ala 463 and Cys 466 buried within the trimer interface of the sulfur-rich core of the TMSs. Our 3D model predicts that the sulfur-rich core of the trimeric BACE1 TMS is accessible to metal ions, but copper ions did not trigger trimerization. The results of functional assays of endogenous BACE1 suggest that it has a role in intracellular copper compartmentalization by transferring cytosolic copper to intracellular compartments, while leaving the overall cellular copper concentration unaltered. Adding to existing physiological models, our results provide novel insight into the atypical interactions between copper and BACE1 and into its non-enzymatic activities. In conclusion, therapeutic Alzheimer disease prevention strategies aimed at decreasing BACE1 protein levels should be regarded with caution, because adverse effects in copper homeostasis may occur. © 2017 by The American Society for Biochemistry and Molecular Biology, Inc.

Effect of Nano-Al₂O₃ on the Toxicity and Oxidative Stress of Copper towards Scenedesmus obliquus.

Science.gov (United States)

Li, Xiaomin; Zhou, Suyang; Fan, Wenhong

2016-06-09

Nano-Al₂O₃ has been widely used in various industries; unfortunately, it can be released into the aquatic environment. Although nano-Al₂O₃ is believed to be of low toxicity, it can interact with other pollutants in water, such as heavy metals. However, the interactions between nano-Al₂O₃ and heavy metals as well as the effect of nano-Al₂O₃ on the toxicity of the metals have been rarely investigated. The current study investigated copper toxicity in the presence of nano-Al₂O₃ towards Scenedesmus obliquus. Superoxide dismutase activity and concentration of glutathione and malondialdehyde in cells were determined in order to quantify oxidative stress in this study. Results showed that the presence of nano-Al₂O₃ reduced the toxicity of Cu towards S. obliquus. The existence of nano-Al₂O₃ decreased the growth inhibition of S. obliquus. The accumulation of copper and the level of oxidative stress in algae were reduced in the presence of nano-Al₂O₃. Furthermore, lower copper accumulation was the main factor that mitigated copper toxicity with the addition of nano-Al₂O₃. The decreased copper uptake could be attributed to the adsorption of copper onto nanoparticles and the subsequent decrease of available copper in water.

Response of the Mediterranean sponge Chondrosia reniformis Nardo to copper pollution

International Nuclear Information System (INIS)

Cebrian, E.; Agell, G.; Marti, R.; Uriz, M.J.

2006-01-01

We examined the effects of exposure to copper pollution on the Atlanto-Mediterranean sponge Chondrosia reniformis. We transplanted sponges from an unpolluted control area to a harbour with a moderately high concentration of copper and measured several biological sponge variables. No effect of this habitat was detected on sponge growth, shape, heat-shock protein expression or metal accumulation. However, a decrease in the clearance rate, an increase in the collagen/cell rate (due to a decrease in the cellular components) and a lower survival rate after 4 months of the sponges transplanted to the harbour was observed. We suggest that copper may alter the sponge physiology, by reducing pumping capacity, which may ultimately lead to sponge death. Consequently, copper pollution exerts strong negative effects on this organism. - Contrasting effects of copper pollution on the Atlanto-Mediterranean sponge Chondrosia reniformis

Microdistribution of copper-carbonate and iron oxide nanoparticles in treated wood

Energy Technology Data Exchange (ETDEWEB)

Matsunaga, Hiroshi, E-mail: mhiroshi@ffpri.affrc.go.jp; Kiguchi, Makoto [Forestry and Forest Products Research Institute (Japan); Evans, Philip D. [University of British Columbia, Centre for Advanced Wood Processing (Canada)

2009-07-15

Aqueous dispersions of copper-carbonate nanoparticles and microparticles have just begun to be exploited commercially for the preservative treatment of wood. The success of the new systems will depend, in part, on the uniform distribution of the preservative in wood and the ability of copper to penetrate cell walls. We examined the distribution of copper in wood treated with a nano-Cu preservative. Copper particles are not uniformly distributed in treated wood, but they accumulate in voids that act as the flow paths for liquids in wood. Particles are deposited on, but not within cell walls. Nevertheless, elemental copper is present within cell walls, but at a lower level than that in wood treated with a conventional wood preservative. These findings suggest that nano-Cu preservatives are able to deliver bioactive components into wood cell walls even though the majority of copper particles are too large to penetrate the cell wall's nanocapillary network.

Divalent Copper as a Major Triggering Agent in Alzheimer's Disease.

Science.gov (United States)

Brewer, George J

2015-01-01

Alzheimer's disease (AD) is at epidemic proportions in developed countries, with a steady increase in the early 1900 s, and then exploding over the last 50 years. This epidemiology points to something causative in the environment of developed countries. This paper will review the considerable evidence that that something could be inorganic copper ingestion. The epidemic parallels closely the spread of copper plumbing, with copper leached from the plumbing into drinking water being a main causal feature, aided by the increasingly common use of supplement pills containing copper. Inorganic copper is divalent copper, or copper-2, while we now know that organic copper, or copper in foods, is primarily monovalent copper, or copper-1. The intestinal transport system, Ctr1, absorbs copper-1 and the copper moves to the liver, where it is put into safe channels. Copper-2 is not absorbed by Ctr1, and some of it bypasses the liver and goes directly into the blood, where it appears to be exquisitely toxic to brain cognition. Thus, while aggregation of amyloid-β has been postulated to be the cause of AD under current dogma, the great increase in prevalence over the last century appears to be due to ingestion of copper-2, which may be causing the aggregation, and/or increasing the oxidant toxicity of the aggregates. An alternative hypothesis proposes that oxidant stress is the primary injuring agent, and under this hypothesis, copper-2 accumulation in the brain may be a causal factor of the oxidant injury. Thus, irrespective of which hypothesis is correct, AD can be classified, at least in part, as a copper-2 toxicity disease. It is relatively easy to avoid copper-2 ingestion, as discussed in this review. If most people begin avoiding copper-2 ingestion, perhaps the epidemic of this serious disease can be aborted.

Copper toxicity in leaves of Elodea canadensis Michx.

Science.gov (United States)

Malec, Przemysław; Maleva, Maria; Prasad, M N V; Strzałka, Kazimierz

2009-05-01

Elodea canadensis (Canadian waterweed) has an ability to accumulate and bioconcentrate heavy metals. In this work, selected cellular responses for Cu treatment were studied in leaves of E. canadensis. Short term experiments, i.e. 1 week exposure to 0.5, 1, 5, and 10 microM of Cu indicated that concentrations up to 10 microM Cu causes a pronounced accumulation of photosynthetic pigments, a drastic degradation of soluble proteins with molecular weight above 18 kDa and a rapid accumulation of polypeptides with molecular weight below 14 kDa. The connection of these observations with copper detoxification mechanisms in aquatic macrophytes are discussed.

Polycyclic aromatic hydrocarbons storage by Fusarium solani in intracellular lipid vesicles

International Nuclear Information System (INIS)

Verdin, Anthony; Lounes-Hadj Sahraoui, Anissa; Newsam, Ray; Robinson, Gary; Durand, Roger

2005-01-01

Accumulation and elimination of polycyclic aromatic hydrocarbons (PAHs) were studied in the fungus Fusarium solani. When the fungus was grown on a synthetic medium containing benzo[a]pyrene, hyphae of F. solani contained numerous lipid vesicles which could be stained by the lipid-specific dyes: Sudan III and Rhodamine B. The fluorescence produced by Rhodamine B and PAH benzo[a]pyrene were at the same locations in the fungal hyphae, indicating that F. solani stored PAH in pre-existing lipid vesicles. A passive temperature-independent process is involved in the benzo[a]pyrene uptake and storage. Sodium azide, a cytochrome c oxidation inhibitor, and the two cytoskeleton inhibitors colchicine and cytochalasin did not prevent the transport and accumulation of PAH in lipid vesicles of F. solani hyphae. F. solani degraded a large range of PAHs at different rates. PAH intracellular storage in lipid vesicles was not necessarily accompanied by degradation and was common to numerous other fungi. - Fungi can store PAHs intracellularly in lipid vesicles independently of their PAH degradation abilities

Polycyclic aromatic hydrocarbons storage by Fusarium solani in intracellular lipid vesicles

Energy Technology Data Exchange (ETDEWEB)

Verdin, Anthony [Laboratoire de Mycologie/Phytopathologie/Environnement, Universite du Littoral-Cote d' Opale, 17 avenue Bleriot, BP 699, 62228 Calais Cedex (France); Lounes-Hadj Sahraoui, Anissa [Laboratoire de Mycologie/Phytopathologie/Environnement, Universite du Littoral-Cote d' Opale, 17 avenue Bleriot, BP 699, 62228 Calais Cedex (France)]. E-mail: lounes@univ-littoral.fr; Newsam, Ray [Department of Biosciences, University of Kent, Canterbury CT2 7NJ (United Kingdom); Robinson, Gary [Department of Biosciences, University of Kent, Canterbury CT2 7NJ (United Kingdom); Durand, Roger [Laboratoire de Mycologie/Phytopathologie/Environnement, Universite du Littoral-Cote d' Opale, 17 avenue Bleriot, BP 699, 62228 Calais Cedex (France)

2005-01-01

Accumulation and elimination of polycyclic aromatic hydrocarbons (PAHs) were studied in the fungus Fusarium solani. When the fungus was grown on a synthetic medium containing benzo[a]pyrene, hyphae of F. solani contained numerous lipid vesicles which could be stained by the lipid-specific dyes: Sudan III and Rhodamine B. The fluorescence produced by Rhodamine B and PAH benzo[a]pyrene were at the same locations in the fungal hyphae, indicating that F. solani stored PAH in pre-existing lipid vesicles. A passive temperature-independent process is involved in the benzo[a]pyrene uptake and storage. Sodium azide, a cytochrome c oxidation inhibitor, and the two cytoskeleton inhibitors colchicine and cytochalasin did not prevent the transport and accumulation of PAH in lipid vesicles of F. solani hyphae. F. solani degraded a large range of PAHs at different rates. PAH intracellular storage in lipid vesicles was not necessarily accompanied by degradation and was common to numerous other fungi. - Fungi can store PAHs intracellularly in lipid vesicles independently of their PAH degradation abilities.

REMOVAL OF COPPER ELECTROLYTE CONTAMINANTS BY ADSORPTION

Directory of Open Access Journals (Sweden)

B Gabai

1997-09-01

Full Text Available Abstract - Selective adsorbents have become frequently used in industrial processes. Recent studies have shown the possibility of using adsorption to separate copper refinery electrolyte contaminants, with better results than those obtained with conventional techniques. During copper electrorefinning, many impurities may be found as dissolved metals present in the anode slime which forms on the electrode surface, accumulated in the electrolyte or incorporated into the refined copper on the cathode by deposition. In this study, synthetic zeolites, chelating resins and activated carbons were tested as adsorbents to select the best adsorbent performance, as well as the best operating temperature for the process. The experimental method applied was the finite bath, which consists in bringing the adsorbent into contact with a finite volume of electrolyte while controlling the temperature. The concentration of metals in the liquid phase was continuously monitored by atomic absorption spectrophotometry (AAS

Metabolism of 25-hydroxyvitamin D in copper-laden rat: A model of Wilson's disease

International Nuclear Information System (INIS)

Carpenter, T.O.; Pendrak, M.L.; Anast, C.S.

1988-01-01

Wilson's disease results in excess tissue accumulation of copper and is often complicated by skeletal and mineral abnormalities. The authors investigated vitamin D metabolism in rats fed a copper-laden diet rendering hepatic copper content comparable with that found in Wilson's disease. Injection of 25-hydroxyvitamin D 3 [25(OH)D 3 ] resulted in reduced 1,25--dihydroxyvitamin D [1,25(OH) 2 D] levels in copper-intoxicated rats. In vitro 25(OH)D-1α-hydroxylase activity was impaired in renal mitochondria from copper-intoxicated animals. Activity was also inhibited in mitochondrial from controls when copper was added to incubation media. Impaired conversion of 25(OH)D to 1,25(OH) 2 D occurs in copper intoxication and suggests that altered vitamin D metabolism is a potential factor in the development of bone and mineral abnormalities in Wilson's disease

Serpula lacrymans, The Dry Rot Fungus and Tolerance Towards Copper-Based Wood Preservatives

DEFF Research Database (Denmark)

Hastrup, Anne Christine Steenkjær; Jensen, Bo; Clausen, Carol

2005-01-01

-rot fungi is thought to be due in part to oxalic acid production and accumulation. Oxalic acid has been implicated in copper tolerance by the formation of copper oxalate crystals. Twelve isolates of the dry rot fungus, S. lacrymans and four other brown rot species were evaluated for weight loss on wood...

Effects of sediment-associated copper to the deposit-feeding snail, Potamopyrgus antipodarum

DEFF Research Database (Denmark)

Pang, Chengfang; Selck, Henriette; Misra, Superb K

2012-01-01

the case of metal oxide NPs using CuO to understand if the effects of NPs differ from micron-sized particles of CuO and aqueous Cu (CuCl2). To address this issue, we compared effects of copper added to the sediment as aqueous Cu, nano- (6 nm) and micro- (... greater effects on growth, feeding rate, and reproduction of P. antipodarum than copper added as micro-CuO or aqueous Cu. P. antipodarum accumulated more copper in the nano-CuO treatment than in aqueous Cu or micro-CuO treatments, indicating that consideration of metal form may be important when assessing...

Biokinetics of copper in black-banded rainbowfish (Melanotaenia nigrans) tolerant to elevated copper concentrations, using the radioisotope 64Cu

International Nuclear Information System (INIS)

Gale, S.; Jeffree, R.; Smith, S.; Lim, R.

2000-01-01

Full text: For over 40 years black-banded rainbowfish (Melanotaenia nigrans) living in the East Branch of the Finniss River, Northern Territory have been exposed to elevated copper concentrations due to mine waste from the Rum Jungle uranium/copper mine. In the 1970s prior to remediation of the mine, fish kills were observed along the length of the East Branch. While copper concentrations remain comparatively high (up to 2000 μg/L) in the East Branch since remediation of the mine site, M. nigrans have been observed in the area. It was, therefore, hypothesised that due to selective pressure of lethal exposure, the population of black-banded rainbowfish in the East Branch have developed a tolerance to elevated copper concentrations. This project aimed to demonstrate copper tolerance and evaluate possible mechanism(s). In May 2000, fish were collected from the East Branch (exposed fish) and from a catchment previously unexposed to elevated metal concentrations (reference fish). The 96-hour EC 50 , fish imbalance (i.e. the concentration of copper that affects 50% of fish over 96 hours) for the exposed fish was over 8 times higher than the reference fish. Using the radioisotope, 64 Cu, the biokinetics of newly accumulated copper was traced in exposed and reference fish at low and elevated copper concentrations. The uptake rate, and therefore body burden, were significantly (p=0.000) lower in exposed fish, at both low and elevated copper concentrations compared to reference fish. Possible mechanisms of reducing copper uptake will be discussed. Tolerance was not lost when fish were maintained in relatively low copper concentrations in the laboratory. Also, the two populations of fish were genetically dissimilar based on allozyme analysis, which suggests that the mechanism is genetically mediated. The outcome of this project will be important in assisting accurate risk assessment and the development of environmental management strategies for the conservation of biota. The

Fabrication of Copper Nanowire Films and their Incorporation into Polymer Matrices for Antibacterial and Marine Antifouling Applications

NARCIS (Netherlands)

Jiang, S.; Sreethawong, T.; Siew Chen Lee, S.; Bee Jin Low, M.; Yin Win, BrzozowskaK.; Brzozowska, A.M.; Lay Ming Teo, S.; Vancso, Gyula J.; Janczewski, D.; Han, M-Y

2015-01-01

With the ban of tributyltin, copper-based biocides are now widely used in antifouling coatings as the major active ingredients. Given the past experience of heavy-metal accumulation in harbors with limited water exchange, there is a significant interest in developing copper materials that greatly

Chaperones CCS, ATOX and COXIV responses to copper supplementation in healthy adults.

Science.gov (United States)

Araya, Magdalena; Andrews, Monica; Pizarro, Fernando; Arredondo, Miguel

2012-04-01

Assessment of proteins in blood and other tissues has failed to identify markers of early copper effects on health. Studies in animal models show that chaperone of SOD (CCS) respond to changes of copper status. Evidence about other copper chaperones (COXIV, ATOX) is not clear. The aim of this study was to assess by means of an in vitro challenge the mRNA relative abundance of ccs, sod1, coxIV, mtIIa and atox in peripheral mononuclear cells (PMNCs) obtained from healthy individuals, acutely and chronically supplemented with small-to-moderate amounts of copper. Healthy participants received 8 mg Cu/d (supplemented group, SG) or placebo, (placebo group, PG) for 2 months. Biochemical indicators were assessed at basal (T0) and after 2 (T2) and 60 days (T60). At these times PMNCs were obtained, challenged with 1, 5 or 20 μM Cu-histidine for 20 h and the mRNA relative abundance of the selected genes assessed by real time PCR. The results showed that at T0, intracellular copper was not different between experimental and control groups. This increased at T2 and T60 when the copper in the media increased (two-way ANOVA, P CCS mRNA transcripts showed no significant changes (two-way ANOVA) at T2 and T60. In SG, CCS changed by treatment, time and interaction (two-way ANOVA, all P CCS but not SOD, ATOX or COXIV responded consistently to controlled changes of copper availability in an in vitro copper challenge.

Fungal accumulation of metals from building materials during brown rot wood decay.

Science.gov (United States)

Hastrup, Anne Christine Steenkjær; Jensen, Bo; Jellison, Jody

2014-08-01

This study analyzes the accumulation and translocation of metal ions in wood during the degradation performed by one strain of each of the three brown rot fungi; Serpula lacrymans, Meruliporia incrassata and Coniophora puteana. These fungi species are inhabitants of the built environment where the prevention and understanding of fungal decay is of high priority. This study focuses on the influence of various building materials in relation to fungal growth and metal uptake. Changes in the concentration of iron, manganese, calcium and copper ions in the decayed wood were analyzed by induced coupled plasma spectroscopy and related to wood weight loss and oxalic acid accumulation. Metal transport into the fungal inoculated wood was found to be dependent on the individual strain/species. The S. lacrymans strain caused a significant increase in total iron whereas the concentration of copper ions in the wood appeared decreased after 10 weeks of decay. Wood inoculated with the M. incrassata isolate showed the contrary tendency with high copper accumulation and low iron increase despite similar weight losses for the two strains. However, significantly lower oxalic acid accumulation was recorded in M. incrassata degraded wood. The addition of a building material resulted in increased weight loss in wood degraded by C. puteana in the soil-block test; however, this could not be directly linked specifically to the accumulation of any of the four metals recorded. The accumulation of oxalic acid seemed to influence the iron uptake. The study assessing the influence of the presence of soil and glass in the soil-block test revealed that soil contributed the majority of the metals for uptake by the fungi and contributed to increased weight loss. The varying uptake observed among the three brown rot fungi strains toward the four metals analyzed may be related to the specific non-enzymatic and enzymatic properties including bio-chelators employed by each of the species during wood

Engineering a predatory bacterium as a proficient killer agent for intracellular bio-products recovery

DEFF Research Database (Denmark)

Martinez, Virginia; Herencias, Cristina; Jurkevitch, Edouard

2016-01-01

This work examines the potential of the predatory bacterium Bdellovibrio bacteriovorus HD100, an obligate predator of other Gram-negative bacteria, as an external cell-lytic agent for recovering valuable intracellular bio-products produced by prey cultures. The bio-product targets to be recovered......% of that accumulated by the prey bacteria, even at high biomass concentrations. This innovative downstream process highlights how B. bacteriovorus can be used as a novel, biological lytic agent for the inexpensive, industrial scale recovery of intracellular products from different Gram-negative prey cultures....

Digitonin abolishes free 2-deoxy-D-glucose accumulation in isolated rat adipocytes

International Nuclear Information System (INIS)

Thompson, K.; Kleinzeller, A.

1986-01-01

The hypothesis that accumulation against sizable chemical gradients of free (non-phosphorylated) 2-deoxy-D-glucose (2dGlc) in isolated rat adipocytes results from an intracellular compartmentation of free hexose was investigated. Cells exposed to 20 μg/ml digitonin for 10' demonstrated an increased plasma membrane permeability indexed by increased L-glucose entry rates and cellular (presumably cytosolic) protein and K + loss. Functional integrity of intracellular organelles was indicated by the ability of the cells to support ATP-driven 45 Ca 2+ -uptake. Equilibrium 3-O-methylglucose (3-O-MG, a non-accumulated hexose) levels were unaffected. These data suggest a specific permeabilizing action of digitonin at the plasma membrane having no effect on intracellular organelles or passively distributed solutes. Upon addition of digitonin, free 2dGlc fell from 66.5 +/- 8.9 to 7.4 +/- 2.3 pmol/10 5 cells, a value not significantly different from 3-O-MG levels. The gradient of 2dGlc-phosphate was also abolished, as was the increased steady-state free 2dGlc levels induced by insulin. The data argue against a compartmentation model as either the mechanism of adipocyte sugar accumulation or the basis of the steady-state free 2dGlc increase seen with insulin and suggest that an intact plasma membrane is essential to the process

Cadmium accumulation by the marine red alga Porphyra umbilicalis

Energy Technology Data Exchange (ETDEWEB)

McLean, M.W.; Williamson, F.B.

1977-01-01

The characteristics of cadmium accumulation by the marine red alga Porphyra umbilicalis L. in culture are reported. The time course of uptake under various light conditions shows that cadmium is concentrated as the result of an on-going anabolic process and not as a consequence of a pH gradient as provided by photosynthesis. The effect of cycloheximide is in agreement with de novo protein synthesis being a prerequisite for cadmium accumulation. Autoradiography suggests a specific intracellular location for bound cadmium--apparently the nucleus.

Adsorption of copper onto char derived macro alga, Undaria pinnatifida

International Nuclear Information System (INIS)

Cho, Hye Jung; Ko, Jeong Huy; Heo, Hyeon Su; Park, Hye Jin; BAe, Yoon Ju; Kim, Jung Hwan; Park, Young-Kwon

2010-01-01

Full text: A release of heavy metals into the environment by industrial activities raises much environmental problems because they tend to remain indefinitely, circulating and eventually accumulating throughout the food chain. Copper is essential to human life and health but, like all heavy metals, is potentially toxic as well. The excessive intakes of copper result in its accumulation in the liver and produce gastrointestinal problems, kidney damage, anemia, and continued inhalation of copper-containing sprays is linked with an increase in lung cancer among exposed people. Consequently, we need to eliminate the copper in drinking water. Also, growth rates of marine macro algae far exceed those of terrestrial biomass, without water limitations, so annual primary production rates are higher for the major marine macro algae than for most terrestrial biomass. According to these reasons, we try to use the macro alga, Undaria pinnatifida. Adsorption of heavy metals is one of the possible technologies involved in the removal of toxic metals from industrial waste streams and mining waste water using low-cost adsorbents. In recent years, many low-cost adsorbents such as seaweeds, activated carbon, etc. have been investigated, but the char by macro alga, Undaria pinnatifida, have not proven to be the most effective and promising substrates. The aim of this study is to remove copper from its aqueous solution by Undaria pinnatifida char for various parameters like pH, contact time, and Cu(II) concentration. The adsorption capacity of Cu(II) by Undaria pinnatifida char was investigated as a function of pH, contact time, and Cu(II) concentration at room temperature. And it was verified using equilibrium studies. (author)

[Structure-functional organization of eukaryotic high-affinity copper importer CTR1 determines its ability to transport copper, silver and cisplatin].

Science.gov (United States)

Skvortsov, A N; Zatulovskiĭ, E A; Puchkova, L V

2012-01-01

It was shown recently, that high affinity Cu(I) importer eukaryotic protein CTR1 can also transport in vitro abiogenic Ag(I) ions and anticancer drug cisplatin. At present there is no rational explanation how CTR1 can transfer platinum group, which is different by coordination properties from highly similar Cu(I) and Ag(I). To understand this phenomenon we analyzed 25 sequences of chordate CTR1 proteins, and found out conserved patterns of organization of N-terminal extracellular part of CTR1 which correspond to initial metal binding. Extracellular copper-binding motifs were qualified by their coordination properties. It was shown that relative position of Met- and His-rich copper-binding motifs in CTR1 predisposes the extracellular CTR1 part to binding of copper, silver and cisplatin. Relation between tissue-specific expression of CTR1 gene, steady-state copper concentration, and silver and platinum accumulation in organs of mice in vivo was analyzed. Significant positive but incomplete correlation exists between these variables. Basing on structural and functional peculiarities of N-terminal part of CTR1 a hypothesis of coupled transport of copper and cisplatin has been suggested, which avoids the disagreement between CTR1-mediated cisplatin transport in vitro, and irreversible binding of platinum to Met-rich peptides.

Hepatocyte nuclear structure and subcellular distribution of copper in zebrafish Brachydanio rerio and roach Rutilus rutilus (Teleostei, Cyprinidae) exposed to copper sulphate

Energy Technology Data Exchange (ETDEWEB)

Paris-Palacios, Severine [Universite de Reims Champagne-Ardenne (URCA), UFR Sciences Exactes et Naturelles, Laboratoire d' Eco-Toxicologie, Institut International de Recherche sur les Ions Metalliques, B.P. 1039-51687 Reims cedex 2 (France)]. E-mail: severine.paris@univ-reims.fr; Biagianti-Risbourg, Sylvie [Universite de Reims Champagne-Ardenne (URCA), UFR Sciences Exactes et Naturelles, Laboratoire d' Eco-Toxicologie, Institut International de Recherche sur les Ions Metalliques, B.P. 1039-51687 Reims cedex 2 (France)]. E-mail: sylvie.biagianti@univ-reims.fr

2006-05-10

Copper is a trace element essential to life, but also a heavy metal with toxic effect clearly demonstrated. Cu induced perturbations in fish liver are well documented but the variability of the reported results is large. In this study two cyprinids, zebrafish and roach, were exposed to copper. Reported histocytological changes are either adaptative or degenerative depending on fish species, concentration of metal, and duration of exposure. Hepatic subcellular distribution of copper was determined by X-ray microanalysis in control and Cu-exposed roach and zebrafish. Sublethal copper sulphate contamination induced the development of a particular nucleolar alteration forming a network or honeycomb like structure in liver. This perturbation is observable in almost all the hepatocytes of zebrafish and roach exposed to copper for a minimum of 4 days of exposure. It seemed to concern more precisely the pars fibrosa. X-ray microanalysis showed that the appearance of network nucleolus was in relation to a Cu accumulation. Cu deposit was well located in the network as pars granulosa and nucloplasm showed very lower metal concentrations. The origin and consequence of network structure in nucleolus was discussed.

Hepatocyte nuclear structure and subcellular distribution of copper in zebrafish Brachydanio rerio and roach Rutilus rutilus (Teleostei, Cyprinidae) exposed to copper sulphate

International Nuclear Information System (INIS)

Paris-Palacios, Severine; Biagianti-Risbourg, Sylvie

2006-01-01

Copper is a trace element essential to life, but also a heavy metal with toxic effect clearly demonstrated. Cu induced perturbations in fish liver are well documented but the variability of the reported results is large. In this study two cyprinids, zebrafish and roach, were exposed to copper. Reported histocytological changes are either adaptative or degenerative depending on fish species, concentration of metal, and duration of exposure. Hepatic subcellular distribution of copper was determined by X-ray microanalysis in control and Cu-exposed roach and zebrafish. Sublethal copper sulphate contamination induced the development of a particular nucleolar alteration forming a network or honeycomb like structure in liver. This perturbation is observable in almost all the hepatocytes of zebrafish and roach exposed to copper for a minimum of 4 days of exposure. It seemed to concern more precisely the pars fibrosa. X-ray microanalysis showed that the appearance of network nucleolus was in relation to a Cu accumulation. Cu deposit was well located in the network as pars granulosa and nucloplasm showed very lower metal concentrations. The origin and consequence of network structure in nucleolus was discussed

Bioindication of air pollution effects near a copper smelter in Brazil using mango trees and soil microbiological properties

International Nuclear Information System (INIS)

Klumpp, Andreas; Hintemann, Therese; Santana Lima, Josanidia; Kandeler, Ellen

2003-01-01

A field study near the copper smelter of a large industrial complex examined air pollution effects on vegetation and soil parameters in Camacari (northeast Brazil). Close to the smelter, soil pH-value was lower and total acidity as well as organic carbon contents were higher compared with a site far from the source and two reference sites. The acidification of top soil particularly and the drastically enhanced plant-available copper concentrations were caused by atmospheric deposition. High sulphur and copper deposition significantly reduced microbial biomass and altered functional diversity of soil microorganisms (arylsulphatase and xylanase). Large accumulations of sulphur, arsenic and copper were detected in mango leaves (Mangifera indica) growing downwind from the smelter suggesting potential food chain-mediated risk. - Atmospheric emissions in northeast Brazil have transformed soil pH, accumulated in soil and plants as sulphur and heavy metals, and affected the functional diversity of soil microorganisms

Bioindication of air pollution effects near a copper smelter in Brazil using mango trees and soil microbiological properties

Energy Technology Data Exchange (ETDEWEB)

Klumpp, Andreas; Hintemann, Therese; Santana Lima, Josanidia; Kandeler, Ellen

2003-12-01

A field study near the copper smelter of a large industrial complex examined air pollution effects on vegetation and soil parameters in Camacari (northeast Brazil). Close to the smelter, soil pH-value was lower and total acidity as well as organic carbon contents were higher compared with a site far from the source and two reference sites. The acidification of top soil particularly and the drastically enhanced plant-available copper concentrations were caused by atmospheric deposition. High sulphur and copper deposition significantly reduced microbial biomass and altered functional diversity of soil microorganisms (arylsulphatase and xylanase). Large accumulations of sulphur, arsenic and copper were detected in mango leaves (Mangifera indica) growing downwind from the smelter suggesting potential food chain-mediated risk. - Atmospheric emissions in northeast Brazil have transformed soil pH, accumulated in soil and plants as sulphur and heavy metals, and affected the functional diversity of soil microorganisms.

Subcellular site and nature of intracellular cadmium in plants

International Nuclear Information System (INIS)

Wagner, G.J.

1979-01-01

The mechanisms underlying heavy metal accumulation, toxicity and tolerance in higher plants are poorly understood. Since subcellular processes are undoubtedly involved in all these phenomena, it is of interest to study the extent of, subcellular site of and nature of intracellularly accumulated cadmium in higher plants. Whole plants supplied 109 CdCl 2 or 112 CdSO 4 accumulated Cd into roots and aerial tissues. Preparation of protoplasts from aerial tissue followed by subcellular fractionation of the protoplasts to obtain intact vacuoles, chloroplasts and cytosol revealed the presence of Cd in the cytosol but not in vacuoles or chloroplasts. Particulate materials containing other cell components were also labeled. Of the 109 Cd supplied to plants, 2 to 10% was recovered in both cytosol preparations and in particulate materials. Cytosol contained proteinaceous--Cd complexes, free metal and low molecular weight Cd complexes. Labeling of protoplasts gave similar results. No evidence was obtained for the production of volatile Cd complexes in tobacco

Dynamic changes in the distribution of minerals in relation to phytic acid accumulation during rice seed development.

Science.gov (United States)

Iwai, Toru; Takahashi, Michiko; Oda, Koshiro; Terada, Yasuko; Yoshida, Kaoru T

2012-12-01

Phytic acid (inositol hexakisphosphate [InsP(6)]) is the storage compound of phosphorus in seeds. As phytic acid binds strongly to metallic cations, it also acts as a storage compound of metals. To understand the mechanisms underlying metal accumulation and localization in relation to phytic acid storage, we applied synchrotron-based x-ray microfluorescence imaging analysis to characterize the simultaneous subcellular distribution of some mineral elements (phosphorus, calcium, potassium, iron, zinc, and copper) in immature and mature rice (Oryza sativa) seeds. This fine-imaging method can reveal whether these elements colocalize. We also determined their accumulation patterns and the changes in phosphate and InsP(6) contents during seed development. While the InsP(6) content in the outer parts of seeds rapidly increased during seed development, the phosphate contents of both the outer and inner parts of seeds remained low. Phosphorus, calcium, potassium, and iron were most abundant in the aleurone layer, and they colocalized throughout seed development. Zinc was broadly distributed from the aleurone layer to the inner endosperm. Copper localized outside the aleurone layer and did not colocalize with phosphorus. From these results, we suggest that phosphorus translocated from source organs was immediately converted to InsP(6) and accumulated in aleurone layer cells and that calcium, potassium, and iron accumulated as phytic acid salt (phytate) in the aleurone layer, whereas zinc bound loosely to InsP(6) and accumulated not only in phytate but also in another storage form. Copper accumulated in the endosperm and may exhibit a storage form other than phytate.

The emerging role of lysosomes in copper homeostasis.

Science.gov (United States)

Polishchuk, Elena V; Polishchuk, Roman S

2016-09-01

The lysosomal system operates as a focal point where a number of important physiological processes such as endocytosis, autophagy and nutrient sensing converge. One of the key functions of lysosomes consists of regulating the metabolism/homeostasis of metals. Metal-containing components are carried to the lysosome through incoming membrane flows, while numerous transporters allow metal ions to move across the lysosome membrane. These properties enable lysosomes to direct metal fluxes to the sites where metal ions are either used by cellular components or sequestered. Copper belongs to a group of metals that are essential for the activity of vitally important enzymes, although it is toxic when in excess. Thus, copper uptake, supply and intracellular compartmentalization have to be tightly regulated. An increasing number of publications have indicated that these processes involve lysosomes. Here we review studies that reveal the expanding role of the lysosomal system as a hub for the control of Cu homeostasis and for the regulation of key Cu-dependent processes in health and disease.

Antioxidant enzymes as bio-markers for copper tolerance in ...

African Journals Online (AJOL)

Safflower (Carthamus tinctorius L.) plants were exposed to 100 μM copper (Cu) for two weeks under controlled environmental conditions using hydroponic culture. Cu induced changes in chlorophyll and carotenoids pigments. More Cu accumulated in the roots as compared with the leaves. Level of lipid peroxidation and ...

The Extracellular Domain of Human High Affinity Copper Transporter (hNdCTR1), Synthesized by E. coli Cells, Chelates Silver and Copper Ions In Vivo.

Science.gov (United States)

Sankova, Tatiana P; Orlov, Iurii A; Saveliev, Andrey N; Kirilenko, Demid A; Babich, Polina S; Brunkov, Pavel N; Puchkova, Ludmila V

2017-11-03

There is much interest in effective copper chelators to correct copper dyshomeostasis in neurodegenerative and oncological diseases. In this study, a recombinant fusion protein for expression in Escherichia coli cells was constructed from glutathione-S-transferase (GST) and the N-terminal domain (ectodomain) of human high affinity copper transporter CTR1 (hNdCTR1), which has three metal-bound motifs. Several biological properties of the GST-hNdCTR1 fusion protein were assessed. It was demonstrated that in cells, the protein was prone to oligomerization, formed inclusion bodies and displayed no toxicity. Treatment of E. coli cells with copper and silver ions reduced cell viability in a dose- and time-dependent manner. Cells expressing GST-hNdCTR1 protein demonstrated resistance to the metal treatments. These cells accumulated silver ions and formed nanoparticles that contained AgCl and metallic silver. In this bacterial population, filamentous bacteria with a length of about 10 µm were often observed. The possibility for the fusion protein carrying extracellular metal binding motifs to integrate into the cell's copper metabolism and its chelating properties are discussed.

Strategies of copper tolerance in root and shoot of broad bean (Vicia faba L.)

International Nuclear Information System (INIS)

Latef, A.A.H.; Alhmad, M.F.A.

2013-01-01

Changes in dry mass and some physiological parameters were investigated in root and shoot of broad bean (Vicia faba L.) exposed to different concentrations of CuSO/sub 4/ (0, 100, 200, 300 and 400 mg kg/sup -1/) for 15 days. Low Cu concentration did not affect dry mass of root and shoot. However, the negative effect of copper on dry mass was noticed in root than shoot especially at the higher concentrations of copper due to the accumulation of Cu was higher in root than that in shoot. Cu induced changes in the concentration of photosynthetic pigments, Zn, K and Ca. Exposure to Cu induced the accumulation of organic solutes (soluble sugar, soluble protein and total free amino acids) in root than shoot. Level of malondialdehyde (MDA) was correlated with the concentration of Cu in broad bean tissues. Copper stimulated the activity of superoxide dismutase (SOD) and peroxidase (POD) and this stimulation was more obvious in shoot than root. (author)

Nano-TiO2 enhances the toxicity of copper in natural water to Daphnia magna

International Nuclear Information System (INIS)

Fan Wenhong; Cui Minming; Liu Hong; Wang Chuan; Shi Zhiwei; Tan Cheng; Yang Xiuping

2011-01-01

The acute toxicity of engineered nanoparticles (NPs) in aquatic environments at high concentrations has been well-established. This study demonstrates that, at a concentration generally considered to be safe in the environment, nano-TiO 2 remarkably enhanced the toxicity of copper to Daphnia magna by increasing the copper bioaccumulation. Specifically, at 2 mg L -1 nano-TiO 2 , the (LC 50 ) of Cu 2+ concentration observed to kill half the population, decreased from 111 μg L -1 to 42 μg L -1 . Correspondingly, the level of metallothionein decreased from 135 μg g -1 wet weight to 99 μg g -1 wet weight at a Cu 2+ level of 100 μg L -1 . The copper was found to be adsorbed onto the nano-TiO 2 , and ingested and accumulated in the animals, thereby causing toxic injury. The nano-TiO 2 may compete for free copper ions with sulfhydryl groups, causing the inhibition of the detoxification by metallothioneins. - Research highlights: → This study demonstrates that, at a concentration generally considered to be safe in the environment, nano-TiO 2 remarkably enhanced the toxicity of copper to Daphnia magna. → The copper was found to be adsorbed onto the nano-TiO 2 , and ingested and accumulated in the Daphnia magna, thereby causing toxic injury. → The nano-TiO 2 may compete for free copper ions with sulfhydryl groups, causing the inhibition of the detoxification mechanism of metallothionein. - The nano-TiO 2 remarkably enhanced the toxicity of copper to Daphnia magna. The nano-TiO 2 may compete for free copper ions with sulfhydryl groups, causing the inhibition of the detoxification mechanism of metallothionein.

Effect of copper and lead on two consortia of phototrophic microorganisms and their capacity to sequester metals

Energy Technology Data Exchange (ETDEWEB)

Burgos, A. [Departament de Genètica i Microbiologia, Universitat Autònoma de Barcelona, Edifici C, Campus de UAB, Bellaterra (Cerdanyola del Vallès), 08193 Barcelona (Spain); Departamento de Recursos Hidrobiológicos, Universidad de Nariño, Pasto (N) (Colombia); Maldonado, J. [Departament de Genètica i Microbiologia, Universitat Autònoma de Barcelona, Edifici C, Campus de UAB, Bellaterra (Cerdanyola del Vallès), 08193 Barcelona (Spain); De los Rios, A. [Museo Nacional de Ciencias Naturales(CSIC), Serrano 115 dpdo, 28006 Madrid (Spain); Solé, A. [Departament de Genètica i Microbiologia, Universitat Autònoma de Barcelona, Edifici C, Campus de UAB, Bellaterra (Cerdanyola del Vallès), 08193 Barcelona (Spain); Esteve, I., E-mail: isabel.esteve@uab.cat [Departament de Genètica i Microbiologia, Universitat Autònoma de Barcelona, Edifici C, Campus de UAB, Bellaterra (Cerdanyola del Vallès), 08193 Barcelona (Spain); Departamento de Recursos Hidrobiológicos, Universidad de Nariño, Pasto (N) (Colombia); Museo Nacional de Ciencias Naturales(CSIC), Serrano 115 dpdo, 28006 Madrid (Spain)

2013-09-15

Highlights: •We studied the tolerance-resistance of phototrophic microorganisms to copper and lead. •We determined the capacity of consortia of microorganisms to sequester copper and lead. •CLSM-λscan is a technique for evaluating in vivo effect of metals on microorganisms. •SEM-EDX and TEM-EDX determined the capacity of microorganisms to sequester metals. -- Abstract: The roles of consortia of phototrophic microorganisms have been investigated in this paper to determine their potential role to tolerate or resist metals and to capture them from polluted cultures. With this purpose, two consortia of microorganisms: on one hand, Geitlerinema sp. DE2011 (Ge) and Scenedesmus sp. DE2009 (Sc) (both identified in this paper by molecular biology methods) isolated from Ebro Delta microbial mats, and on the other, Spirulina sp. PCC 6313 (Sp) and Chroococcus sp. PCC 9106 (Ch), from Pasteur culture collection were polluted with copper and lead. In order to analyze the ability of these consortia to tolerate and capture metals, copper and lead were selected, because both have been detected in Ebro Delta microbial mats. The tolerance-resistance to copper and lead for both consortia was determined in vivo and at cellular level by Confocal Laser Scanning Microscopy (CLSM-λscan function). The results obtained demonstrate that both consortia are highly tolerant-resistant to lead and that the limits between the copper concentration having cytotoxic effect and that having an essential effect are very close in these microorganisms. The capacity of both consortia to capture extra- and intracellular copper and lead was determined by Scanning Electron Microscopy (SEM) and Transmission Electron Microscopy (TEM) respectively, coupled to an Energy Dispersive X-ray detector (EDX). The results showed that all the microorganisms assayed were able to capture copper extracellularly in the extrapolymeric substances, and lead extra- and intracellularly in polyphosphate inclusions. Moreover

Effect of copper and lead on two consortia of phototrophic microorganisms and their capacity to sequester metals

International Nuclear Information System (INIS)

Burgos, A.; Maldonado, J.; De los Rios, A.; Solé, A.; Esteve, I.

2013-01-01

Highlights: •We studied the tolerance-resistance of phototrophic microorganisms to copper and lead. •We determined the capacity of consortia of microorganisms to sequester copper and lead. •CLSM-λscan is a technique for evaluating in vivo effect of metals on microorganisms. •SEM-EDX and TEM-EDX determined the capacity of microorganisms to sequester metals. -- Abstract: The roles of consortia of phototrophic microorganisms have been investigated in this paper to determine their potential role to tolerate or resist metals and to capture them from polluted cultures. With this purpose, two consortia of microorganisms: on one hand, Geitlerinema sp. DE2011 (Ge) and Scenedesmus sp. DE2009 (Sc) (both identified in this paper by molecular biology methods) isolated from Ebro Delta microbial mats, and on the other, Spirulina sp. PCC 6313 (Sp) and Chroococcus sp. PCC 9106 (Ch), from Pasteur culture collection were polluted with copper and lead. In order to analyze the ability of these consortia to tolerate and capture metals, copper and lead were selected, because both have been detected in Ebro Delta microbial mats. The tolerance-resistance to copper and lead for both consortia was determined in vivo and at cellular level by Confocal Laser Scanning Microscopy (CLSM-λscan function). The results obtained demonstrate that both consortia are highly tolerant-resistant to lead and that the limits between the copper concentration having cytotoxic effect and that having an essential effect are very close in these microorganisms. The capacity of both consortia to capture extra- and intracellular copper and lead was determined by Scanning Electron Microscopy (SEM) and Transmission Electron Microscopy (TEM) respectively, coupled to an Energy Dispersive X-ray detector (EDX). The results showed that all the microorganisms assayed were able to capture copper extracellularly in the extrapolymeric substances, and lead extra- and intracellularly in polyphosphate inclusions. Moreover

Electrochemical Behavior of Pure Copper in Phosphate Buffer Solutions: A Comparison Between Micro- and Nano-Grained Copper

Science.gov (United States)

Imantalab, O.; Fattah-alhosseini, A.; Keshavarz, M. K.; Mazaheri, Y.

2016-02-01

In this work, electrochemical behavior of annealed (micro-) and nano-grained pure copper (fabricated by accumulative roll bonding process) in phosphate buffer solutions of various pH values ranging from 10.69 to 12.59 has been studied. Before any electrochemical measurements, evaluation of microstructure was obtained by optical microscope and transmission electron microscopy. To investigate the electrochemical behavior of the samples, the potentiodynamic polarization, Mott-Schottky analysis, and electrochemical impedance spectroscopy (EIS) were carried out. Potentiodynamic polarization plots and EIS measurements revealed that as a result of grain refinement, the passive behavior of the nano-grained sample was improved compared to that of annealed pure copper. Also, Mott-Schottky analysis indicated that the passive films behaved as p-type semiconductors and grain refinement did not change the semiconductor type of passive films.

Intracellular transport of low density lipoprotein-derived cholesterol is defective in Niemann-Pick type C fibroblasts

International Nuclear Information System (INIS)

Liscum, L.; Ruggiero, R.M.; Faust, J.R.

1989-01-01

Niemann-Pick disease type C (NPC) is characterized by substantial intracellular accumulation of unesterified cholesterol. The accumulation of unesterified cholesterol in NPC fibroblasts cultured with low density lipoprotein (LDL) appears to result from the inability of LDL to stimulate cholesterol esterification in addition to impaired LDL-mediated downregulation of LDL receptor activity and cellular cholesterol synthesis. Although a defect in cholesterol transport in NPC cells has been inferred from previous studies, no experiments have been reported that measure the intracellular movement of LDL-cholesterol specifically. We have used four approaches to assess intracellular cholesterol transport in normal and NPC cells and have determined the following: (a) mevinolin-inhibited NPC cells are defective in using LDL-cholesterol for growth. However, exogenously added mevalonate restores cell growth equally in normal and NPC cells; (b) the transport of LDL-derived [3H]cholesterol to the plasma membrane is slower in NPC cells, while the rate of appearance of [3H]acetate-derived, endogenously synthesized [3H]cholesterol at the plasma membrane is the same for normal and NPC cells; (c) in NPC cells, LDL-derived [3H]cholesterol accumulates in lysosomes to higher levels than normal, resulting in defective movement to other cell membranes; and (d) incubation of cells with LDL causes an increase in cholesterol content of NPC lysosomes that is threefold greater than that observed in normal lysosomes. Our results indicate that a cholesterol transport defect exists in NPC that is specific for LDL-derived cholesterol

Effect of Nano-Al2O3 on the Toxicity and Oxidative Stress of Copper towards Scenedesmus obliquus

Science.gov (United States)

Li, Xiaomin; Zhou, Suyang; Fan, Wenhong

2016-01-01

Nano-Al2O3 has been widely used in various industries; unfortunately, it can be released into the aquatic environment. Although nano-Al2O3 is believed to be of low toxicity, it can interact with other pollutants in water, such as heavy metals. However, the interactions between nano-Al2O3 and heavy metals as well as the effect of nano-Al2O3 on the toxicity of the metals have been rarely investigated. The current study investigated copper toxicity in the presence of nano-Al2O3 towards Scenedesmus obliquus. Superoxide dismutase activity and concentration of glutathione and malondialdehyde in cells were determined in order to quantify oxidative stress in this study. Results showed that the presence of nano-Al2O3 reduced the toxicity of Cu towards S. obliquus. The existence of nano-Al2O3 decreased the growth inhibition of S. obliquus. The accumulation of copper and the level of oxidative stress in algae were reduced in the presence of nano-Al2O3. Furthermore, lower copper accumulation was the main factor that mitigated copper toxicity with the addition of nano-Al2O3. The decreased copper uptake could be attributed to the adsorption of copper onto nanoparticles and the subsequent decrease of available copper in water. PMID:27294942

LDL Receptors as Gateways for Intracellular Porphyrin Uptake

International Nuclear Information System (INIS)

Novick, S.; Laster, B.; Quastel, M.

2004-01-01

Boronated compounds are currently being studied for possible use in Boron Neutron Capture Therapy (BNCT). We found that one of these agents, BOPP (tetrakis-carborane-carboxylate, esters of 2,4-bis (a,b- dihydroxyethyl) deuteroporphyrin IX), could also be labeled with indium (In-BOPP) and, therefore, could also be used potentially to transport high Z atoms into tumor cell DNA for AET (Auger Electron Therapy). In order to assess the uptake of these agents into cells, the role of the LDL receptor in the intracellular accumulation of BOPP and In-BOPP was investigated. Pre-incubation of V-79 Chinese hamster cells in medium containing delipidized fetal bovine serum (FBS) markedly increased the subsequent uptake of intracellular boron transported by both BOPP and In-BOPP when compared with cells that had been pre-incubated with medium containing 10% normal FBS (lipidized). The increased uptake was characterized by elevated levels of receptor, and greater affinity was shown for both BOPP and In-BOPP, although less marked with the latter. Positive cooperativity was demonstrated by sigmoid saturation curves, Scatchard analysis and Hill plots. Increasing the amount of LDL in the incubation medium had a relatively small effect on the total accumulation of either indium or boron atoms inside the cell. Furthermore, chemical acetylation of LDL did not decrease the intracellular uptake of either boron or indium transported by BOPP or In-BOPP. It is thus concluded that BOPP and In-BOPP preferentially enter the cells directly by way of the LDL receptor and that only a small fraction of these molecules are transported into the cells indirectly using serum LDLs as their carriers. These data suggest a novel way of bringing greater amounts of boron and indium (and perhaps other agents) into tissues. Porphyrins can be used to transport different agents into tumor cells because they are tumor affinic molecules. Tumors express a higher number of LDL receptors than do most normal tissues

Accumulation of hepatic Hsp70 and plasma cortisol in Oreochromis ...

African Journals Online (AJOL)

The hepatic isoforms Hsp70, Hsp74 and Hsp76 were identified and quantified from copper exposures. Long-term DDT exposure did not result in significant induction of hepatic Hsp70. An increase in plasma cortisol concentration was associated with a decrease in heat shock protein accumulation after cadmium exposure, ...

Overexpression of amyloid precursor protein increases copper content in HEK293 cells

International Nuclear Information System (INIS)

Suazo, Miriam; Hodar, Christian; Morgan, Carlos; Cerpa, Waldo; Cambiazo, Veronica; Inestrosa, Nibaldo C.; Gonzalez, Mauricio

2009-01-01

Amyloid precursor protein (APP) is a transmembrane glycoprotein widely expressed in mammalian tissues and plays a central role in Alzheimer's disease. However, its physiological function remains elusive. Cu 2+ binding and reduction activities have been described in the extracellular APP135-156 region, which might be relevant for cellular copper uptake and homeostasis. Here, we assessed Cu 2+ reduction and 64 Cu uptake in two human HEK293 cell lines overexpressing APP. Our results indicate that Cu 2+ reduction increased and cells accumulated larger levels of copper, maintaining cell viability at supra-physiological levels of Cu 2+ ions. Moreover, wild-type cells exposed to both Cu 2+ ions and APP135-155 synthetic peptides increased copper reduction and uptake. Complementation of function studies in human APP751 transformed Fre1 defective Saccharomyces cerevisiae cells rescued low Cu 2+ reductase activity and increased 64 Cu uptake. We conclude that Cu 2+ reduction activity of APP facilitates copper uptake and may represent an early step in cellular copper homeostasis.

Intracellular Cholesterol Trafficking and Impact in Neurodegeneration

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Fabian Arenas

2017-11-01

Full Text Available Cholesterol is a critical component of membrane bilayers where it plays key structural and functional roles by regulating the activity of diverse signaling platforms and pathways. Particularly enriched in brain, cholesterol homeostasis in this organ is singular with respect to other tissues and exhibits a heterogeneous regulation in distinct brain cell populations. Due to the key role of cholesterol in brain physiology and function, alterations in cholesterol homeostasis and levels have been linked to brain diseases and neurodegeneration. In the case of Alzheimer disease (AD, however, this association remains unclear with evidence indicating that either increased or decreased total brain cholesterol levels contribute to this major neurodegenerative disease. Here, rather than analyzing the role of total cholesterol levels in neurodegeneration, we focus on the contribution of intracellular cholesterol pools, particularly in endolysosomes and mitochondria through its trafficking via specialized membrane domains delineated by the contacts between endoplasmic reticulum and mitochondria, in the onset of prevalent neurodegenerative diseases such as AD, Parkinson disease, and Huntington disease as well as in lysosomal disorders like Niemann-Pick type C disease. We dissect molecular events associated with intracellular cholesterol accumulation, especially in mitochondria, an event that results in impaired mitochondrial antioxidant defense and function. A better understanding of the mechanisms involved in the distribution of cholesterol in intracellular compartments may shed light on the role of cholesterol homeostasis disruption in neurodegeneration and may pave the way for specific intervention opportunities.

Soil and sediment concentrations of chromium, copper, and arsenic adjacent to a chromated copper arsenate-treated wetland boardwalk

Science.gov (United States)

Stan Lebow; Daniel Foster

2010-01-01

Environmental accumulation of preservative adjacent to a chromated copper arsenate (type C)–treated wetland boardwalk was evaluated. The site is considered a realistic ‘‘worst case’’ because of the large volume of treated wood, low current speeds, high annual rainfall, and environmental sensitivity. Soil and sediment samples were collected before construction and 0.5,...

Characterization of Lactobacillus brevis L62 strain, highly tolerant to copper ions.

Science.gov (United States)

Mrvčić, Jasna; Butorac, Ana; Solić, Ema; Stanzer, Damir; Bačun-Družina, Višnja; Cindrić, Mario; Stehlik-Tomas, Vesna

2013-01-01

Lactic acid bacteria (LAB) as starter culture in food industry must be suitable for large-scale industrial production and possess the ability to survive in unfavorable processes and storage conditions. Approaches taken to address these problems include the selection of stress-resistant strains. In food industry, LAB are often exposed to metal ions induced stress. The interactions between LAB and metal ions are very poorly investigated. Because of that, the influence of non-toxic, toxic and antioxidant metal ions (Zn, Cu, and Mn) on growth, acid production, metal ions binding capacity of wild and adapted species of Leuconostoc mesenteroides L3, Lactobacillus brevis L62 and Lactobacillus plantarum L73 were investigated. The proteomic approach was applied to clarify how the LAB cells, especially the adapted ones, protect themselves and tolerate high concentrations of toxic metal ions. Results have shown that Zn and Mn addition into MRS medium in the investigated concentrations did not have effect on the bacterial growth and acid production, while copper ions were highly toxic, especially in static conditions. Leuc. mesenteroides L3 was the most efficient in Zn binding processes among the chosen LAB species, while L. plantarum L73 accumulated the highest concentration of Mn. L. brevis L62 was the most copper resistant species. Adaptation had a positive effect on growth and acid production of all species in the presence of copper. However, the adapted species incorporated less metal ions than the wild species. The exception was adapted L. brevis L62 that accumulated high concentration of copper ions in static conditions. The obtained results showed that L. brevis L62 is highly tolerant to copper ions, which allows its use as starter culture in fermentative processes in media with high concentration of copper ions.

Lipophilic ester and amide derivatives of rosmarinic acid protect cells against H2O2-induced DNA damage and apoptosis: The potential role of intracellular accumulation and labile iron chelation

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Paraskevi S. Gerogianni

2018-05-01

Full Text Available Phenolic acids represent abundant components contained in human diet. However, the negative charge in their carboxylic group limits their capacity to diffuse through biological membranes, thus hindering their access to cell interior. In order to promote the diffusion of rosmarinic acid through biological membranes, we synthesized several lipophilic ester- and amide-derivatives of this compound and evaluated their capacity to prevent H2O2-induced DNA damage and apoptosis in cultured human cells. Esterification of the carboxylic moiety with lipophilic groups strongly enhanced the capacity of rosmarinic acid to protect cells. On the other hand, the amide-derivatives were somewhat less effective but exerted less cytotoxicity at high concentrations. Cell uptake experiments, using ultra-high performance liquid chromatography coupled with tandem mass spectrometry (UHPLC-MS/MS, illustrated different levels of intracellular accumulation among the ester- and amide-derivatives, with the first being more effectively accumulated, probably due to their extensive hydrolysis inside the cells. In conclusion, these results highlight the hitherto unrecognized fundamental importance of derivatization of diet-derived phenolic acids to unveil their biological potential.

Highly Efficient Intracellular Protein Delivery by Cationic Polyethyleneimine-Modified Gelatin Nanoparticles

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Ming-Ju Chou

2018-02-01

Full Text Available Intracellular protein delivery may provide a safe and non-genome integrated strategy for targeting abnormal or specific cells for applications in cell reprogramming therapy. Thus, highly efficient intracellular functional protein delivery would be beneficial for protein drug discovery. In this study, we generated a cationic polyethyleneimine (PEI-modified gelatin nanoparticle and evaluated its intracellular protein delivery ability in vitro and in vivo. The experimental results showed that the PEI-modified gelatin nanoparticle had a zeta potential of approximately +60 mV and the particle size was approximately 135 nm. The particle was stable at different biological pH values and temperatures and high protein loading efficiency was observed. The fluorescent image results revealed that large numbers of particles were taken up into the mammalian cells and escaped from the endosomes into the cytoplasm. In a mouse C26 cell-xenograft cancer model, particles accumulated in cancer cells. In conclusion, the PEI-modified gelatin particle may provide a biodegradable and highly efficient protein delivery system for use in regenerative medicine and cancer therapy.

Effect of copper and lead on two consortia of phototrophic microorganisms and their capacity to sequester metals.

Science.gov (United States)

Burgos, A; Maldonado, J; De Los Rios, A; Solé, A; Esteve, I

2013-09-15

The roles of consortia of phototrophic microorganisms have been investigated in this paper to determine their potential role to tolerate or resist metals and to capture them from polluted cultures. With this purpose, two consortia of microorganisms: on one hand, Geitlerinema sp. DE2011 (Ge) and Scenedesmus sp. DE2009 (Sc) (both identified in this paper by molecular biology methods) isolated from Ebro Delta microbial mats, and on the other, Spirulina sp. PCC 6313 (Sp) and Chroococcus sp. PCC 9106 (Ch), from Pasteur culture collection were polluted with copper and lead. In order to analyze the ability of these consortia to tolerate and capture metals, copper and lead were selected, because both have been detected in Ebro Delta microbial mats. The tolerance-resistance to copper and lead for both consortia was determined in vivo and at cellular level by Confocal Laser Scanning Microscopy (CLSM-λscan function). The results obtained demonstrate that both consortia are highly tolerant-resistant to lead and that the limits between the copper concentration having cytotoxic effect and that having an essential effect are very close in these microorganisms. The capacity of both consortia to capture extra- and intracellular copper and lead was determined by Scanning Electron Microscopy (SEM) and Transmission Electron Microscopy (TEM) respectively, coupled to an Energy Dispersive X-ray detector (EDX). The results showed that all the microorganisms assayed were able to capture copper extracellularly in the extrapolymeric substances, and lead extra- and intracellularly in polyphosphate inclusions. Moreover, the studied micro-organisms did not exert any inhibitory effect on each other's metal binding capacity. From the results obtained in this paper, it can be concluded that consortia of phototrophic microorganisms could play a very important role in biorepairing sediments polluted by metals, as a result of their ability to tolerate or resist high concentrations of metals and to

Intracellular accumulation of potent amiloride analogues by human neutrophils

International Nuclear Information System (INIS)

Simchowitz, L.; Woltersdorf, O.W. Jr.; Cragoe, E.J. Jr.

1987-01-01

The mechanism of uptake of a series of amiloride derivatives by human neutrophils was investigated using [ 14 C]amiloride and the 14 C-labeled 5-(1-hexahydroazepinyl)-6-bromo analogue (BrMM) which is approximately 500-fold more potent than the parent compound at inhibiting Na+/H+ exchange. At an external concentration of 2 microM, the influx of BrMM at 37 degrees C was rapid, reaching a steady state by approximately 20 min. The rate of BrMM uptake (approximately 25 mumol/liter.min) was approximately 90-fold faster than for the same concentration of amiloride, a finding which correlates with differences in lipid partitioning of the two compounds. Uptake was unrelated to specific binding to Na+/H+ exchange transport sites: influx of either drug was nonsaturable whereas amiloride- and BrMM-mediated inhibition of Na+/H+ countertransport obeyed Michaelis-Menten kinetics with apparent Ki values of approximately 75 and approximately 0.2 microM. Entry occurred exclusively via the neutral (uncharged) forms (pK'a 8.40-8.55). Influx was markedly pH-dependent: it was enhanced by extracellular alkalinization and reduced by acidification. Influx was, however, insensitive to large changes in membrane voltage, thereby implying the protonated (charged) species to be impermeant. About 75% of the total intracellular pool of amiloride, but only approximately 25% of BrMM, is contained within the lysosomes, an expected consequence of the partitioning and subsequent trapping of a weak base within this strongly acidic subcellular compartment. With BrMM, there was a relative approximately 60-fold enrichment in the internal/external water concentration ratio of the drug; the value for amiloride was much less, approximately 4. This disparity is consistent with substantial binding of BrMM to internal constituents, presumably to proteins and/or nucleic acids

TOR (target of rapamycin) is a key regulator of triacylglycerol accumulation in microalgae.

Science.gov (United States)

Imamura, Sousuke; Kawase, Yasuko; Kobayashi, Ikki; Shimojima, Mie; Ohta, Hiroyuki; Tanaka, Kan

2016-01-01

Most microalgae abundantly accumulate lipid droplets (LDs) containing triacylglycerols (TAGs) under several stress conditions, but the underlying molecular mechanism of this accumulation remains unclear. In a recent study, we found that inhibition of TOR (target of rapamycin), a highly conserved protein kinase of eukaryotes, by rapamycin resulted in TAG accumulation in microalgae, indicating that TOR negatively regulates TAG accumulation. Here, we show that formation of intracellular LDs and TAG accumulation were also induced in the unicellular green alga Chlamydomonas reinhardtii after exposure to Torin1 or AZD8055, which are novel TOR inhibitors that inhibit TOR activity in a manner different from rapamycin. These results supported quite well our previous conclusion that TOR is a central regulator of TAG accumulation in microalgae.

An econometric model of the U.S. secondary copper industry: Recycling versus disposal

Science.gov (United States)

Slade, M.E.

1980-01-01

In this paper, a theoretical model of secondary recovery is developed that integrates microeconomic theories of production and cost with a dynamic model of scrap generation and accumulation. The model equations are estimated for the U.S. secondary copper industry and used to assess the impacts that various policies and future events have on copper recycling rates. The alternatives considered are: subsidies for secondary production, differing energy costs, and varying ore quality in primary production. ?? 1990.

MUC1 intra-cellular trafficking is clathrin, dynamin, and rab5 dependent

International Nuclear Information System (INIS)

Liu Xiaolong; Yuan Zhenglong; Chung, Maureen

2008-01-01

MUC1, a transmembrane glycoprotein, is abnormally over-expressed in most human adenocarcinomas. MUC1 association with cytoplasmic cell signal regulators and nuclear accumulation are important for its tumor related activities. Little is known about how MUC1 translocates from the cell membrane to the cytoplasm. In this study, live cell imaging was used to study MUC1 intracellular trafficking. The interaction between EGFR and MUC1 was mapped by FRET analysis and EGF stimulated MUC1 endocytosis was observed directly through live cell imaging. MUC1-CT endocytosis was clathrin and dynamin dependent. Rab5 over-expression resulted in decreased cell membrane localization of MUC1, with accumulation of MUC1 endocytic vesicles in the peri-nuclear region. Conversely, over-expression of a Rab5 dominant negative mutant (S34N) resulted in redistribution of MUC1 from the peri-nuclear region to the cytoplasm. Collectively, these results indicated that MUC1 intra-cellular trafficking occurs through a regulated process that was stimulated by direct EGFR and MUC1 interaction, mediated by clathrin coated pits that were dynamin dependent and regulated by Rab5

Synthesis of cytochrome c oxidase 1 (SCO1) inhibits insulin sensitivity by decreasing copper levels in adipocytes.

Science.gov (United States)

Wei, Xiang-Bo; Guo, Liang; Liu, Yang; Zhou, Shui-Rong; Liu, Yuan; Dou, Xin; Du, Shao-Yue; Ding, Meng; Peng, Wan-Qiu; Qian, Shu-Wen; Huang, Hai-Yan; Tang, Qi-Qun

2017-09-23

Dysregulation of insulin signaling leads to type 2 diabetes mellitus (T2DM) and other metabolic disorders. Obesity is an important contributor to insulin resistance, and although the understanding of this relationship has improved in recent years, the mechanism of obesity-induced insulin resistance is not completely understood. Disorders of copper metabolism tend to accompany the development of obesity, which increases the risk of insulin resistance. Synthesis of cytochrome c oxidase 1 (SCO1) functions in the assembly of cytochrome c oxidase (COX) and cellular copper homeostasis. However, the role of SCO1 in the regulation of metabolism remains unknown. Here, we found that obese mice had higher expression of SCO1 and lower levels of copper in white adipose tissue (WAT) than did the control mice. Overexpression of SCO1 in adipocytes was associated with copper deficiency. Copper increased insulin sensitivity by decreasing the level of phosphatase and tensin homolog (PTEN) protein. Ectopic expression of SCO1 led to insulin resistance and was accompanied by a decrease in intracellular copper level, and addition of copper abolished the inhibitory effect of SCO1 on insulin sensitivity. Our results demonstrated a novel role of SCO1 in modulating insulin sensitivity via the regulation of copper concentration in WAT and suggested a potential therapeutic target for T2DM. Copyright © 2017. Published by Elsevier Inc.

The Extracellular Domain of Human High Affinity Copper Transporter (hNdCTR1, Synthesized by E. coli Cells, Chelates Silver and Copper Ions In Vivo

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Tatiana P. Sankova

2017-11-01

Full Text Available There is much interest in effective copper chelators to correct copper dyshomeostasis in neurodegenerative and oncological diseases. In this study, a recombinant fusion protein for expression in Escherichia coli cells was constructed from glutathione-S-transferase (GST and the N-terminal domain (ectodomain of human high affinity copper transporter CTR1 (hNdCTR1, which has three metal-bound motifs. Several biological properties of the GST-hNdCTR1 fusion protein were assessed. It was demonstrated that in cells, the protein was prone to oligomerization, formed inclusion bodies and displayed no toxicity. Treatment of E. coli cells with copper and silver ions reduced cell viability in a dose- and time-dependent manner. Cells expressing GST-hNdCTR1 protein demonstrated resistance to the metal treatments. These cells accumulated silver ions and formed nanoparticles that contained AgCl and metallic silver. In this bacterial population, filamentous bacteria with a length of about 10 µm were often observed. The possibility for the fusion protein carrying extracellular metal binding motifs to integrate into the cell’s copper metabolism and its chelating properties are discussed.

Comparison of intracellular accumulation and cytotoxicity of free mTHPC and mTHPC-loaded PLGA nanoparticles in human colon carcinoma cells

International Nuclear Information System (INIS)

Loew, Karin; Wagner, Sylvia; Briesen, Hagen von; Knobloch, Thomas; Wiehe, Arno; Engel, Andrea; Langer, Klaus

2011-01-01

The second generation photosensitizer mTHPC was approved by the European Medicines Agency (EMA) for the palliative treatment of advanced head and neck cancer in October 2001. It is known that mTHPC possesses a significant phototoxicity against a variety of human cancer cells in vitro but also exhibits dark toxicity and can cause adverse effects (especially skin photosensitization). Due to its poor water solubility, the administration of hydrophobic photosensitizer still presents several difficulties. To overcome the administration problems, the use of nanoparticles as drug carrier systems is much investigated. Nanoparticles based on poly(lactic-co-glycolic acid) (PLGA) have been extensively studied as delivery systems into tumours due to their biocompatibility and biodegradability. The goal of this study was the comparison of free mTHPC and mTHPC-loaded PLGA nanoparticles concerning cytotoxicity and intracellular accumulation in human colon carcinoma cells (HT29). The nanoparticles delivered the photosensitizer to the colon carcinoma cells and enabled drug release without losing its activity. The cytotoxicity assays showed a time- and concentration-dependent decrease in cell proliferation and viability after illumination. However, first and foremost mTHPC lost its dark toxic effects using the PLGA nanoparticles as a drug carrier system. Therefore, PLGA nanoparticles are a promising drug carrier system for the hydrophobic photosensitizer mTHPC.

CPB1 of Aedes aegypti Interacts with DENV2 E Protein and Regulates Intracellular Viral Accumulation and Release from Midgut Cells

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Hong-Wai Tham

2014-12-01

Full Text Available Aedes aegypti is a principal vector responsible for the transmission of dengue viruses (DENV. To date, vector control remains the key option for dengue disease management. To develop new vector control strategies, a more comprehensive understanding of the biological interactions between DENV and Ae. aegypti is required. In this study, a cDNA library derived from the midgut of female adult Ae. aegypti was used in yeast two-hybrid (Y2H screenings against DENV2 envelope (E protein. Among the many interacting proteins identified, carboxypeptidase B1 (CPB1 was selected, and its biological interaction with E protein in Ae. aegypti primary midgut cells was further validated. Our double immunofluorescent assay showed that CPB1-E interaction occurred in the endoplasmic reticulum (ER of the Ae. aegypti primary midgut cells. Overexpression of CPB1 in mosquito cells resulted in intracellular DENV2 genomic RNA or virus particle accumulation, with a lower amount of virus release. Therefore, we postulated that in Ae. aegypti midgut cells, CPB1 binds to the E protein deposited on the ER intraluminal membranes and inhibits DENV2 RNA encapsulation, thus inhibiting budding from the ER, and may interfere with immature virus transportation to the trans-Golgi network.

Comparison of intracellular accumulation and cytotoxicity of free mTHPC and mTHPC-loaded PLGA nanoparticles in human colon carcinoma cells

Energy Technology Data Exchange (ETDEWEB)

Loew, Karin; Wagner, Sylvia; Briesen, Hagen von [Fraunhofer-Institute for Biomedical Engineering, D-66386 Strasse Ingbert (Germany); Knobloch, Thomas [Institute of Pharmaceutical Technology, Biocenter of Goethe-University, D-60438 Frankfurt (Germany); Wiehe, Arno [Biolitec AG, D-07745 Jena (Germany); Engel, Andrea; Langer, Klaus, E-mail: hagen.briesen@ibmt.fraunhofer.de [Institute of Pharmaceutical Technology and Biopharmacy, University of Muenster, D-48149 Muenster (Germany)

2011-06-17

The second generation photosensitizer mTHPC was approved by the European Medicines Agency (EMA) for the palliative treatment of advanced head and neck cancer in October 2001. It is known that mTHPC possesses a significant phototoxicity against a variety of human cancer cells in vitro but also exhibits dark toxicity and can cause adverse effects (especially skin photosensitization). Due to its poor water solubility, the administration of hydrophobic photosensitizer still presents several difficulties. To overcome the administration problems, the use of nanoparticles as drug carrier systems is much investigated. Nanoparticles based on poly(lactic-co-glycolic acid) (PLGA) have been extensively studied as delivery systems into tumours due to their biocompatibility and biodegradability. The goal of this study was the comparison of free mTHPC and mTHPC-loaded PLGA nanoparticles concerning cytotoxicity and intracellular accumulation in human colon carcinoma cells (HT29). The nanoparticles delivered the photosensitizer to the colon carcinoma cells and enabled drug release without losing its activity. The cytotoxicity assays showed a time- and concentration-dependent decrease in cell proliferation and viability after illumination. However, first and foremost mTHPC lost its dark toxic effects using the PLGA nanoparticles as a drug carrier system. Therefore, PLGA nanoparticles are a promising drug carrier system for the hydrophobic photosensitizer mTHPC.

An Evaluation of Kinetic Parameters of Cadmium and Copper Biosorption by Immobilized Cells

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Nelly Georgieva

2007-10-01

Full Text Available Bioremediation is the use of living organisms to reduce or eliminate environmental hazards resulting from the accumulation of toxic chemicals and other hazardous wastes. This technology is based on the utilization of microorganisms to transform organic and inorganic compounds. The filamentous yeast Trichosporon cutaneum strain R57, immobilized and free cells was cultivated as batch culture on a liquid medium in the presence of various concentrations of cadmium and copper ions. The simultaneous uptake and accumulation of Cd2+ and Cu2+ ions by Tr. cutaneum cells depending on the initial concentration of Cd2+ and Cu2+ in the medium were studied. The potential use of the free and immobilized cells of Trichosporon cutaneum to remove cadmium and copper ions, from aqueous solutions was evaluated. Two important physicochemical aspects for the evaluation of the sorption process as a unit operation are the equilibrium of sorption and the kinetics. The Cd2+ and Cu2+ ions biosorption capacities of all tested adsorbent were presented as a function of the initial concentration of metal ions within the aqueous biosorption medium. The individual, as well as bicomponent sorption kinetics of copper and cadmium ions by immobilised cells of Trichosporon cutaneum R57 is presented. A second order kinetic model obtains kinetic parameters for the copper and cadmium ions.

Investigation of heavy metal accumulation in Polygonum thunbergii for phytoextraction

International Nuclear Information System (INIS)

Kim, In Sung; Kang, Kyung Hong; Johnson-Green, Perry; Lee, Eun Ju

2003-01-01

Polygonum thunbergii is useful to remove heavy metal from soil and water. - In this study, cadmium (II), lead (II), copper (II) and zinc (II) were determined in Polygonum thunbergii and soil from the Mankyung River watershed, Korea. Soil samples contained detectable lead ( -1 ), copper ( -1 ) and zinc ( -1 ), whereas cadmium was undetectable. Whole plants of P. thunbergii contained detectable lead ( -1 ), copper ( -1 ) and zinc ( -1 ), whereas cadmium was detectable only in the stem ( -1 ) and root ( -1 ). Whole plant concentrations were very different for each metal, particularly in the case of zinc. The mean content of heavy metal in the whole plants increased in the order of cadmium (8.5 μg g -1 ) -1 ) -1 ) -1 ). Soil lead, copper and zinc were correlated with each metal's accumulation in the plants (lead, r=0.841, P<0.005; copper, r=0.874, P<0.001; zinc, r=0.770, P<0.005). Lead content in roots and leaves was highly correlated (r=0.5529, P<0.001), as was lead content in roots and stems (r=0.5425, P<0.001). Mean bioconcentration factors for the aboveground tissues were 4.2 (lead), 14.8 (copper) and 27.7 (zinc), and for the underground tissues, were 22.2 (lead), 92.9 (copper) and 62.7 (zinc). After hydroponic growth, bioaccumulation coefficients were 2.0 (cadmium), 3.2 (lead), 17.2 (copper) and 13.1 (zinc) for whole plants. We considered these results as indicative of the ability of P. thunbergii plants to take up metal ions from a soil matrix contaminated with heavy metals

Gama-aminobutyric acid accumulation in Elsholtzia splendens in response to copper toxicity*

OpenAIRE

Yang, Xiao-e; Peng, Hong-yun; Tian, Sheng-ke

2005-01-01

A solution with different Cu supply levels was cultured to investigate gama-aminobutyric acid (GABA) accumulation in Elsholtzia splendens, a native Chinese Cu-tolerant and accumulating plant species. Increasing Cu from 0.25 to 500 μmol/L significantly enhanced levels of GABA and histidine (His), but considerably decreased levels of aspartate (Asp) and glutamate (Glu) in the leaves. The leaf Asp level negatively correlated with leaf Cu level, while leaf GABA level positively correlated with le...

Interaction between copper and radiocesium in Indian mustard and sunflower grown in the hydroponic solution

International Nuclear Information System (INIS)

Shirong Tang; Xiaochang Wang

2002-01-01

Both Indian mustard and sunflower were grown in a hydroponic solution treated with different concentration activities of 134 Cs or with different amounts of copper or with both in order to investigate the interaction between copper and radiocesium. It was found that 134 Cs activity concentration applied in the nutrient solution exerted more influence on the uptake and translocation of copper by Indian mustard than by sunflower. Indian mustard grown in hydroponic solution containing certain levels of copper and being treated with higher 134 Cs activity concentration showed higher uptake of copper than sunflower. However, in the case of root copper concentrations, sunflower showed significantly higher copper immobilization by roots than Indian mustard. It was also found that the presence of copper the the hydroponic solution did modify radiocesium uptake by both species. The application of 1 mg/l in the growth medium could greatly increase the uptake of 134 Cs by both species. With 3 mg/l concentration of copper amended to the solution, the accumulation of 134 Cs by both species was decreased compared to the 1 mg/l copper treatment. These lines of evidence show that there is stronger interaction between copper and radiocesium in Indian mustard than in sunflower during the root uptake through nutrient solution. (author)

Intracellular Retention of ABL Kinase Inhibitors Determines Commitment to Apoptosis in CML Cells

Science.gov (United States)

Dziadosz, Marek; Schnöder, Tina; Heidel, Florian; Schemionek, Mirle; Melo, Junia V.; Kindler, Thomas; Müller-Tidow, Carsten; Koschmieder, Steffen; Fischer, Thomas

2012-01-01

Clinical development of imatinib in CML established continuous target inhibition as a paradigm for successful tyrosine kinase inhibitor (TKI) therapy. However, recent reports suggested that transient potent target inhibition of BCR-ABL by high-dose TKI (HD-TKI) pulse-exposure is sufficient to irreversibly commit cells to apoptosis. Here, we report a novel mechanism of prolonged intracellular TKI activity upon HD-TKI pulse-exposure (imatinib, dasatinib) in BCR-ABL-positive cells. Comprehensive mechanistic exploration revealed dramatic intracellular accumulation of TKIs which closely correlated with induction of apoptosis. Cells were rescued from apoptosis upon HD-TKI pulse either by repetitive drug wash-out or by overexpression of ABC-family drug transporters. Inhibition of ABCB1 restored sensitivity to HD-TKI pulse-exposure. Thus, our data provide evidence that intracellular drug retention crucially determines biological activity of imatinib and dasatinib. These studies may refine our current thinking on critical requirements of TKI dose and duration of target inhibition for biological activity of TKIs. PMID:22815843

Separation of copper-64 from copper phthalocyanine

International Nuclear Information System (INIS)

Battaglin, R.I.M.

1979-01-01

The separation of copper-64 from irradiated copper phthalocyanine by Szilard-Chalmers effect is studied. Two methods of separation are used: one of them is based on the dissolution of the irradiated dry compound in concentrated sulfuric acid following its precipitation in water. In the other one the compound is irradiated with water in paste form following treatment with water and hydrochloric acid. The influence of the crystal form of the copper phthalocyanine on the separation yield of copper-64 is shown. Preliminary tests using the ionic exchange technique for purification and changing of copper-64 sulfate to chloride form are carried out. The specific activity using the spectrophotometric technique, after the determination of the copper concentration in solution of copper-64, is calculated. (Author) [pt

Lipid Accumulation in Peripheral Blood Dendritic Cells and Anticancer Immunity in Patients with Lung Cancer

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Ryo Arai

2018-01-01

Full Text Available We studied the subsets of peripheral blood dendritic cells (DCs and lipid accumulation in DCs to investigate the involvement of DCs in the decreased anticancer immunity of advanced lung cancer patients. We analyzed the population of DC subsets in peripheral blood using flow cytometry. We then determined lipid accumulation in the DCs using BODIPY 650/665, a fluorophore with an affinity for lipids. Compared with healthy controls, the number of DCs in the peripheral blood of treatment-naive cancer patients was significantly reduced. In patients with stage III + IV disease, the numbers of myeloid DCs (mDCs and plasmacytoid DCs were also significantly reduced. Lipid accumulation in DCs evaluated based on the fluorescence intensity of BODIPY 650/665 was significantly higher in stage III + IV lung cancer patients than in the controls. In the subset analysis, the fluorescence was highest for mDCs. The intracellularly accumulated lipids were identified as triglycerides. A decreased mixed leukocyte reaction was observed in the mDCs from lung cancer patients compared with those from controls. Taken together, the results show that lung cancer patients have a notably decreased number of peripheral blood DCs and their function as antigen-presenting cells is decreased due to their high intracellular lipid accumulation. Thereby, anticancer immunity is suppressed.

Caesium accumulation by microorganisms: uptake mechanisms, cation competition, compartmentalization and toxicity

International Nuclear Information System (INIS)

Avery, S.V.

1995-01-01

The continued release of caesium radioisotopes into the environment has led to a resurgence of interest in microbe-Cs interactions. Caesium exists almost exclusively as the monovalent cation Cs + in the natural environment. Although Cs + is a weak Lewis acid that exhibits a low tendency to form complexes with ligands, its chemical similarity to the biologically essential alkali cation K + facilitates high levels of metabolism-dependent intracellular accumulation. Microbial Cs + (K + ) uptake is generally mediated by monovalent cation transport systems located on the plasma membrane. These differ widely in specificity for alkali cations and consequently microorganisms display large differences in their ability to accumulate Cs + ; Cs + appears to have an equal or greater affinity than K + for transport in certain microorganisms. Microbial Cs + accumulation is markedly influenced by the presence of external cations, e.g. K + , Na + , NH 4 + and H + , and is generally accompanied by an approximate stoichiometric exchange for intracellular K + . However, stimulation of growth of K + -starved microbial cultures by Cs + is limited and it has been proposed that it is not the presence of Cs + in cells that is growth inhibitory but rather the resulting loss of K + . Increased microbial tolerance to Cs + may result from sequestration of Cs + in vacuoles or changes in the activity and/or specificity of transport systems mediating Cs + uptake. The precise intracellular target(s) for Cs + - induced toxicity has yet to be clearly defined, although certain internal structures, e.g. ribosomes, become unstable in the presence of Cs + and Cs + is known to substitute poorly for K + in the activation of many K + -requiring enzymes. (author)

Accumulation of Trace Metal Elements (Cu, Zn, Cd, and Pb in Surface Sediment via Decomposed Seagrass Leaves: A Mesocosm Experiment Using Zostera marina L.

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Shinya Hosokawa

Full Text Available Accumulation of Cu, Zn, Cd, and Pb in the sediment of seagrass ecosystems was examined using mesocosm experiments containing Zostera marina (eelgrass and reference pools. Lead was approximately 20-fold higher in the surface sediment in the eelgrass pool than in eelgrass leaves and epiphytes on the eelgrass leaves, whereas zinc and cadmium were significantly lower in the surface sediment than in the leaves, with intermediate concentrations in epiphytes. Copper concentrations were similar in both the surface sediment and leaves but significantly lower in epiphytes. Carbon and nitrogen contents increased significantly with increasing δ13C in surface sediments of both the eelgrass and reference pools. Copper, Zn, Cd, and Pb also increased significantly with increasing δ13C in the surface sediment in the eelgrass pool but not in the reference pool. By decomposition of eelgrass leaves with epiphytes, which was examined in the eelgrass pool, copper and lead concentrations increased more than 2-fold and approximately a 10-fold, whereas zinc and cadmium concentrations decreased. The high copper and lead concentrations in the surface sediment result from accumulation in decomposed, shed leaves, whereas zinc and cadmium remobilized from decomposed shed leaves but may remain at higher concentrations in the leaves than in the original sediments. The results of our mesocosm study demonstrate that whether the accumulation or remobilization of trace metals during the decomposition of seagrass leaves is trace metal dependent, and that the decomposed seagrass leaves can cause copper and lead accumulation in sediments in seagrass ecosystems.

Overexpression of rice glutaredoxin OsGrx_C7 and OsGrx_C2.1 reduces intracellular arsenic accumulation and increases tolerance in Arabidopsis thaliana

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Pankaj Kumar Verma

2016-06-01

Full Text Available Glutaredoxins (Grxs are a family of small multifunctional proteins involved in various cellular functions, including redox regulation and protection under oxidative stress. Despite the high number of Grx genes in plant genomes (48 Grxs in rice, the biological functions and physiological roles of most of them remain unknown. Here, the functional characterization of the two arsenic-responsive rice Grx family proteins, OsGrx_C7 and OsGrx_C2.1 are reported. Over-expression of OsGrx_C7 and OsGrx_C2.1 in transgenic Arabidopsis thaliana conferred arsenic (As tolerance as reflected by germination, root growth assay, and whole plant growth. Also, the transgenic expression of OsGrxs displayed significantly reduced As accumulation in A. thaliana seeds and shoot tissues compared to WT plants during both AsIII and AsV stress. Thus, OsGrx_C7 and OsGrx_C2.1 seem to be an important determinant of As-stress response in plants. OsGrx_C7 and OsGrx_C2.1 transgenic showed to maintain intracellular GSH pool and involved in lowering AsIII accumulation either by extrusion or reducing uptake by altering the transcript of A. thaliana AtNIPs. Overall, OsGrx_C7 and OsGrx_C2.1 may represent a Grx family protein involved in As stress response and may allow a better understanding of the As induced stress pathways and the design of strategies for the improvement of stress tolerance as well as decreased As content in crops.

Lead, zinc and copper accumulation and tolerance in populations of Paspalum distichum and Cynodon dactylon

International Nuclear Information System (INIS)

Shu, W.S.; Ye, Z.H.; Lan, C.Y.; Zhang, Z.Q.; Wong, M.H.

2002-01-01

Metal-tolerant populations of the plants Paspalum distichum and Cunodon dactylon were identified. - Both Fankou and Lechang lead/zinc (Pb/Zn) mine tailings located at Guangdong Province contained high levels of total and DTPA-extractable Pb, Zn and Cu. Paspalum distichum and Cynodon dactylon were dominant species colonized naturally on the tailings. Lead, zinc and copper accumulation and tolerance of different populations of the two grasses growing on the tailings were investigated. Tillers of these populations including those from an uncontaminated area were subjected to the following concentrations: 5, 10, 20, 30 and 40 mg l -1 Pb, 2.5, 5, 10, 20 and 30 mg l -1 Zn, or 0.25, 0.50, 1 and 2 mg l -1 Cu for 14 days, respectively, then tolerance index (TI) and EC 50 (the concentrations of metals in solutions which reduce 50% of normal root growth) were calculated. The results indicated that both Lechang and Fankou populations of the two grasses showed a greater tolerance to the three metals than those growing on the uncontaminated area, which suggested that co-tolerant ecotypes have evolved in the two grasses. P. distichum collected from Fankou tailings had the highest tolerance to Cu while Lechang population the highest tolerance to Pb and Zn among the tested populations, and tolerance levels in P. distichum were related to metal concentrations in the plants. P. distichum had a better growth performance than C. dactylon when both of them were grown on the tailings sites. Tolerant populations of these species would serve as potential candidates for re-vegetation of wastelands contaminated with Pb, Zn and Cu

Lead, zinc and copper accumulation and tolerance in populations of Paspalum distichum and Cynodon dactylon

Energy Technology Data Exchange (ETDEWEB)

Shu, W.S.; Ye, Z.H.; Lan, C.Y.; Zhang, Z.Q.; Wong, M.H

2002-12-01

Metal-tolerant populations of the plants Paspalum distichum and Cunodon dactylon were identified. - Both Fankou and Lechang lead/zinc (Pb/Zn) mine tailings located at Guangdong Province contained high levels of total and DTPA-extractable Pb, Zn and Cu. Paspalum distichum and Cynodon dactylon were dominant species colonized naturally on the tailings. Lead, zinc and copper accumulation and tolerance of different populations of the two grasses growing on the tailings were investigated. Tillers of these populations including those from an uncontaminated area were subjected to the following concentrations: 5, 10, 20, 30 and 40 mg l{sup -1} Pb, 2.5, 5, 10, 20 and 30 mg l{sup -1} Zn, or 0.25, 0.50, 1 and 2 mg l{sup -1} Cu for 14 days, respectively, then tolerance index (TI) and EC{sub 50} (the concentrations of metals in solutions which reduce 50% of normal root growth) were calculated. The results indicated that both Lechang and Fankou populations of the two grasses showed a greater tolerance to the three metals than those growing on the uncontaminated area, which suggested that co-tolerant ecotypes have evolved in the two grasses. P. distichum collected from Fankou tailings had the highest tolerance to Cu while Lechang population the highest tolerance to Pb and Zn among the tested populations, and tolerance levels in P. distichum were related to metal concentrations in the plants. P. distichum had a better growth performance than C. dactylon when both of them were grown on the tailings sites. Tolerant populations of these species would serve as potential candidates for re-vegetation of wastelands contaminated with Pb, Zn and Cu.

Anti fouling effect of two saturated copper coatings applied on carbon steel structures

International Nuclear Information System (INIS)

Guiamet, P. S.; Gomez de Saravia, S. G.

2008-01-01

Biofouling is the colonization of man-made substrata by sessile organisms. The aim of this paper is to evaluate the performance of two anti fouling saturated copper coating. Bioassays were carried out at a harbor in Argentine (38 degree centigrade 02' S-57 degree centigrade 32'W). during six months, one series of pipes and panels were removed monthly to estimate the recruitment of macro and micro fouling species and immediately replaced by clean ones. Another series was removed from the beginning of exposure to monitor the development of the established community (accumulative pipes and panels along six months). Data obtained from control (without a saturated copper coating) and saturated-copper coated pipes and panels were compared in order to estimate performance of the coating. One of two saturated copper coating demonstrated a good effect anti fouling. (Author) 25 refs

Copper in the sediment and sea surface microlayer near a fallowed, open-net fish farm.

Science.gov (United States)

Loucks, Ronald H; Smith, Ruth E; Fisher, Clyde V; Fisher, E Brian

2012-09-01

Sediment and sea surface microlayer samples near an open-net salmon farm in Nova Scotia, were analysed for copper. Copper is a constituent of the feed and is an active ingredient of anti-foulants. The salmon farm was placed in fallow after 15 years of production. Sampling was pursued over 27 months. Elevated copper concentrations in the sediments indicated the farm site as a source. Bubble flotation due to gas-emitting sediments from eutrophication is a likely process for accumulating copper in the sea surface microlayer at enriched concentrations. Elevated and enriched concentrations in the sea surface microlayer over distance from the farm site led, as a result of wind-drift, to an enlarged farm footprint. The levels of copper in both sediments and sea surface microlayer exceeded guidelines for protection of marine life. Over the 27 months period, copper levels persisted in the sediments and decreased gradually in the sea surface microlayer. Copyright © 2012 Elsevier Ltd. All rights reserved.

Utilization of Copper Alloys for Marine Applications

Science.gov (United States)

Drach, Andrew

Utilization of copper alloy components in systems deployed in marine environment presents potential improvements by reducing maintenance costs, prolonging service life, and increasing reliability. However, integration of these materials faces technological challenges, which are discussed and addressed in this work, including characterization of material performance in seawater environment, hydrodynamics of copper alloy components, and design procedures for systems with copper alloys. To characterize the hydrodynamic behavior of copper alloy nets, mesh geometry of the major types of copper nets currently used in the marine aquaculture are analyzed and formulae for the solidity and strand length are proposed. Experimental studies of drag forces on copper alloy net panels are described. Based on these studies, empirical values for normal drag coefficients are proposed for various types of copper netting. These findings are compared to the previously published data on polymer nets. It is shown that copper nets exhibit significantly lower resistance to normal currents, which corresponds to lower values of normal drag coefficient. The seawater performance (corrosion and biofouling) of copper alloys is studied through the field trials of tensioned and untensioned specimens in a one-year deployment in the North Atlantic Ocean. The corrosion behavior is characterized by weight loss, optical microscopy, and SEM/EDX analyses. The biofouling performance is quantified in terms of the biomass accumulation. To estimate the effects of stray electrical currents on the seawater corrosion measurements, a low cost three-axis stray electric current monitoring device is designed and tested both in the lab and in the 30-day field deployment. The system consists of a remotely operated PC with a set of pseudo-electrodes and a digital compass. The collected data is processed to determine magnitudes of AC and DC components of electric field and dominant AC frequencies. Mechanical behavior of

Trichloroethylene-mediated cytotoxicity in human epidermal keratinocytes is mediated by the rapid accumulation of intracellular calcium: Interception by naringenin.

Science.gov (United States)

Ali, F; Khan, A Q; Khan, R; Sultana, S

2016-02-01

Industrial solvents pose a significant threat to the humankind. The mechanisms of their toxicity still remain in debate. Trichloroethylene (TCE) is a widespread industrial solvent responsible for severe liver dysfunction, cutaneous toxicity in occupationally exposed humans. We utilized an in vitro system of human epidermal keratinocyte (HaCaT) cells in this study to avoid complex cell and extracellular interactions. We report the cytotoxicity of organic solvent TCE in HaCaT and its reversal by a natural flavanone, naringenin (Nar). The cytotoxicity was attributed to the rapid intracellular free calcium (Ca(2+)) release, which might lead to the elevation of protein kinase C along with robust free radical generation, instability due to energy depletion, and sensitization of intracellular stress signal transducer nuclear factor κB. These effects were actually seen to induce significant amount of genomic DNA fragmentation. Furthermore, all these effects of TCE were effectively reversed by the treatment of Nar, a natural flavanone. Our studies identify intracellular Ca as a unique target used by organic solvents in the cytotoxicity and highlight the Ca(2+) ion stabilizer properties of Nar. © The Author(s) 2015.

Intracellular Nitrate of Marine Diatoms as a Driver of Anaerobic Nitrogen Cycling in Sinking Aggregates

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Anja Kamp

2016-11-01

Full Text Available Diatom-bacteria aggregates are key for the vertical transport of organic carbon in the ocean. Sinking aggregates also represent pelagic microniches with intensified microbial activity, oxygen depletion in the center, and anaerobic nitrogen cycling. Since some of the aggregate-forming diatom species store nitrate intracellularly, we explored the fate of intracellular nitrate and its availability for microbial metabolism within anoxic diatom-bacteria aggregates. The ubiquitous nitrate-storing diatom Skeletonema marinoi was studied as both axenic cultures and laboratory-produced diatom-bacteria aggregates. Stable 15N isotope incubations under dark and anoxic conditions revealed that axenic S. marinoi is able to reduce intracellular nitrate to ammonium that is immediately excreted by the cells. When exposed to a light:dark cycle and oxic conditions, S. marinoi stored nitrate intracellularly in concentrations > 60 mmol L-1 both as free-living cells and associated to aggregates. Intracellular nitrate concentrations exceeded extracellular concentrations by three orders of magnitude. Intracellular nitrate was used up within 2-3 days after shifting diatom-bacteria aggregates to dark and anoxic conditions. Thirty-one percent of the diatom-derived nitrate was converted to nitrogen gas, indicating that a substantial fraction of the intracellular nitrate pool of S. marinoi becomes available to the aggregate-associated bacterial community. Only 5% of the intracellular nitrate was reduced to ammonium, while 59% was recovered as nitrite. Hence, aggregate-associated diatoms accumulate nitrate from the surrounding water and sustain complex nitrogen transformations, including loss of fixed nitrogen, in anoxic, pelagic microniches. Additionally, it may be expected that intracellular nitrate not converted before the aggregates have settled onto the seafloor could fuel benthic nitrogen transformations.

Interactions between Metal-binding Domains Modulate Intracellular Targeting of Cu(I)-ATPase ATP7B, as Revealed by Nanobody Binding*

Science.gov (United States)

Huang, Yiping; Nokhrin, Sergiy; Hassanzadeh-Ghassabeh, Gholamreza; Yu, Corey H.; Yang, Haojun; Barry, Amanda N.; Tonelli, Marco; Markley, John L.; Muyldermans, Serge; Dmitriev, Oleg Y.; Lutsenko, Svetlana

2014-01-01

The biologically and clinically important membrane transporters are challenging proteins to study because of their low level of expression, multidomain structure, and complex molecular dynamics that underlies their activity. ATP7B is a copper transporter that traffics between the intracellular compartments in response to copper elevation. The N-terminal domain of ATP7B (N-ATP7B) is involved in binding copper, but the role of this domain in trafficking is controversial. To clarify the role of N-ATP7B, we generated nanobodies that interact with ATP7B in vitro and in cells. In solution NMR studies, nanobodies revealed the spatial organization of N-ATP7B by detecting transient functionally relevant interactions between metal-binding domains 1–3. Modulation of these interactions by nanobodies in cells enhanced relocalization of the endogenous ATP7B toward the plasma membrane linking molecular and cellular dynamics of the transporter. Stimulation of ATP7B trafficking by nanobodies in the absence of elevated copper provides direct evidence for the important role of N-ATP7B structural dynamics in regulation of ATP7B localization in a cell. PMID:25253690

Apoplastic and intracellular plant sugars regulate developmental transitions in witches’ broom disease of cacao

Science.gov (United States)

Barau, Joan; Grandis, Adriana; Carvalho, Vinicius Miessler de Andrade; Teixeira, Gleidson Silva; Zaparoli, Gustavo Henrique Alcalá; do Rio, Maria Carolina Scatolin; Rincones, Johana; Buckeridge, Marcos Silveira; Pereira, Gonçalo Amarante Guimarães

2015-01-01

Witches’ broom disease (WBD) of cacao differs from other typical hemibiotrophic plant diseases by its unusually long biotrophic phase. Plant carbon sources have been proposed to regulate WBD developmental transitions; however, nothing is known about their availability at the plant–fungus interface, the apoplastic fluid of cacao. Data are provided supporting a role for the dynamics of soluble carbon in the apoplastic fluid in prompting the end of the biotrophic phase of infection. Carbon depletion and the consequent fungal sensing of starvation were identified as key signalling factors at the apoplast. MpNEP2, a fungal effector of host necrosis, was found to be up-regulated in an autophagic-like response to carbon starvation in vitro. In addition, the in vivo artificial manipulation of carbon availability in the apoplastic fluid considerably modulated both its expression and plant necrosis rate. Strikingly, infected cacao tissues accumulated intracellular hexoses, and showed stunted photosynthesis and the up-regulation of senescence markers immediately prior to the transition to the necrotrophic phase. These opposite findings of carbon depletion and accumulation in different host cell compartments are discussed within the frame of WBD development. A model is suggested to explain phase transition as a synergic outcome of fungal-related factors released upon sensing of extracellular carbon starvation, and an early senescence of infected tissues probably triggered by intracellular sugar accumulation. PMID:25540440

Evaluation of Intracellular Signaling Downstream Chimeric Antigen Receptors.

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Hannah Karlsson

Full Text Available CD19-targeting CAR T cells have shown potency in clinical trials targeting B cell leukemia. Although mainly second generation (2G CARs carrying CD28 or 4-1BB have been investigated in patients, preclinical studies suggest that third generation (3G CARs with both CD28 and 4-1BB have enhanced capacity. However, little is known about the intracellular signaling pathways downstream of CARs. In the present work, we have analyzed the signaling capacity post antigen stimulation in both 2G and 3G CARs. 3G CAR T cells expanded better than 2G CAR T cells upon repeated stimulation with IL-2 and autologous B cells. An antigen-driven accumulation of CAR+ cells was evident post antigen stimulation. The cytotoxicity of both 2G and 3G CAR T cells was maintained by repeated stimulation. The phosphorylation status of intracellular signaling proteins post antigen stimulation showed that 3G CAR T cells had a higher activation status than 2G. Several proteins involved in signaling downstream the TCR were activated, as were proteins involved in the cell cycle, cell adhesion and exocytosis. In conclusion, 3G CAR T cells had a higher degree of intracellular signaling activity than 2G CARs which may explain the increased proliferative capacity seen in 3G CAR T cells. The study also indicates that there may be other signaling pathways to consider when designing or evaluating new generations of CARs.

Copper and copper-nickel alloys as zebra mussel antifoulants

Energy Technology Data Exchange (ETDEWEB)

Dormon, J.M.; Cottrell, C.M.; Allen, D.G.; Ackerman, J.D.; Spelt, J.K. [Univ. of Toronto, Ontario (Canada)

1996-04-01

Copper has been used in the marine environment for decades as cladding on ships and pipes to prevent biofouling by marine mussels (Mytilus edulis L.). This motivated the present investigation into the possibility of using copper to prevent biofouling in freshwater by both zebra mussels and quagga mussels (Dreissena polymorpha and D. bugensis collectively referred to as zebra mussels). Copper and copper alloy sheet proved to be highly effective in preventing biofouling by zebra mussels over a three-year period. Further studies were conducted with copper and copper-nickel mesh (lattice of expanded metal) and screen (woven wire with a smaller hole size), which reduced the amount of copper used. Copper screen was also found to be strongly biofouling-resistant with respect to zebra mussels, while copper mesh reduced zebra mussel biofouling in comparison to controls, but did not prevent it entirely. Preliminary investigations into the mechanism of copper antifouling, using galvanic couples, indicated that the release of copper ions from the surface of the exposed metal into the surrounding water is directly or indirectly responsible for the biofouling resistance of copper.

Chronic copper exposure and fatty acid composition of the amphipod Dikerogammarus villosus: Results from a field study

Energy Technology Data Exchange (ETDEWEB)

Maazouzi, Chafik [Universite de Metz, Laboratoire Interactions Ecotoxicologie Biodiversite Ecosystemes (LIEBE), CNRS UMR 7146, Avenue General Delestraint, 57070 Metz (France)], E-mail: maazouzi@univ-metz.fr; Masson, Gerard [Universite de Metz, Laboratoire Interactions Ecotoxicologie Biodiversite Ecosystemes (LIEBE), CNRS UMR 7146, Avenue General Delestraint, 57070 Metz (France)], E-mail: masson1@univ-metz.fr; Izquierdo, Maria Soledad [Grupo de Investigacion en Acuicultura, ULPGC and ICCM, P.O. Box 56, 35200 Telde, Las Palmas de Gran Canaria (Spain)], E-mail: mizquierdo@dbio.ulpgc.es; Pihan, Jean-Claude [Universite de Metz, Laboratoire Interactions Ecotoxicologie Biodiversite Ecosystemes (LIEBE), CNRS UMR 7146, Avenue General Delestraint, 57070 Metz (France)], E-mail: pihan@univ-metz.fr

2008-11-15

Field study allows assessment of long-term effects on fatty acid (FA) composition of organisms under chronic exposure to metals. One expected effect of copper is peroxidation of lipids and essentially polyunsaturated fatty acids (PUFA). FA analysis was established for the amphipod Dikerogammarus villosus subjected to different degrees of copper exposure (4-40 {mu}g Cu L{sup -1}). A previous study in our team showed that this species regulates its body Cu concentration (106-135 mg Cu kg{sup -1} dry weight). Despite the high capacity of bioaccumulation, the absence of a correlation between copper concentration in D. villosus and water prevents its use as bioindicator of copper pollution. Both sexes from the most polluted site showed the lowest total FA content, but the highest PUFA percent, mainly of the long-chained variety (C20-C22). Mechanisms leading to the prevention of lipid peroxidation in this species were discussed (metallothioneins and intracellular granules) and proposed with support from literature data. - Under chronic copper exposure, Dikerogammarus villosus loses in total fatty acids content but increases its essential {omega}3 and {omega}6 PUFA percent.

Metal X-ray microanalysis in the olfactory system of rainbow trout exposed to low level of copper

International Nuclear Information System (INIS)

Julliard, A.K.; Astic, L.; Saucier, D.

1995-01-01

It has recently been shown that a chronic copper exposure induces specific degeneration of olfactory receptor cells in rainbow trout; however, the exact mechanism of action of the metal is not yet known. Using X-ray microanalysis in transmission electron microscopy, we have studied the distribution of metal in the olfactory system of fish exposed for 15,30 and 60 days to 20 μg/l of copper. This was done in order to determine if it was accumulated in receptor cells and transported into the central nervous system via the olfactory nerve. No copper accumulation was detected either in the olfactory epithelium, in the olfactory nerve or in the olfactory bulb. The heavy metal was exclusively found within melanosomes of melanophores located in the lamina propria. After 60 days of exposure, the copper content in melanosomes was about two-fold higher than that in controls. As far as some morphological recovery took place in the olfactory organ during the metal exposure, which lets us suppose that some detoxication mechanism occurs, it could be suggested that metanophores might be somehow involved in such a mechanism. (authors). 57 refs., 15 figs

Accumulation, subcellular distribution and toxicity of inorganic mercury and methylmercury in marine phytoplankton

Energy Technology Data Exchange (ETDEWEB)

Wu Yun [Division of Life Science, Hong Kong University of Science and Technology (HKUST), Clear Water Bay, Kowloon (Hong Kong); Wang Wenxiong, E-mail: wwang@ust.hk [Division of Life Science, Hong Kong University of Science and Technology (HKUST), Clear Water Bay, Kowloon (Hong Kong)

2011-10-15

We examined the accumulation, subcellular distribution, and toxicity of Hg(II) and MeHg in three marine phytoplankton (the diatom Thalassiosira pseudonana, the green alga Chlorella autotrophica, and the flagellate Isochrysis galbana). For MeHg, the inter-species toxic difference could be best interpreted by the total cellular or intracellular accumulation. For Hg(II), both I. galbana and T. pseudonana exhibited similar sensitivity, but they each accumulated a different level of Hg(II). A higher percentage of Hg(II) was bound to the cellular debris fraction in T. pseudonana than in I. galbana, implying that the cellular debris may play an important role in Hg(II) detoxification. Furthermore, heat-stable proteins were a major binding pool for MeHg, while the cellular debris was an important binding pool for Hg(II). Elucidating the different subcellular fates of Hg(II) and MeHg may help us understand their toxicity in marine phytoplankton at the bottom of aquatic food chains. - Highlights: > The inter-species toxic difference of methylmercury in marine phytoplankton can be explained by its total cellular or intracellular accumulation. > The inter-species toxic difference of inorganic mercury in marine phytoplankton can be explained by its subcellular distribution. > Heat-stable protein was a major binding pool for MeHg, while the cellular debris was an important binding pool for Hg(II). - The inter-species difference in methylmercury and inorganic mercury toxicity in phytoplankton can be explained by cellular accumulation and subcellular distribution.

Accumulation, subcellular distribution and toxicity of inorganic mercury and methylmercury in marine phytoplankton

International Nuclear Information System (INIS)

Wu Yun; Wang Wenxiong

2011-01-01

We examined the accumulation, subcellular distribution, and toxicity of Hg(II) and MeHg in three marine phytoplankton (the diatom Thalassiosira pseudonana, the green alga Chlorella autotrophica, and the flagellate Isochrysis galbana). For MeHg, the inter-species toxic difference could be best interpreted by the total cellular or intracellular accumulation. For Hg(II), both I. galbana and T. pseudonana exhibited similar sensitivity, but they each accumulated a different level of Hg(II). A higher percentage of Hg(II) was bound to the cellular debris fraction in T. pseudonana than in I. galbana, implying that the cellular debris may play an important role in Hg(II) detoxification. Furthermore, heat-stable proteins were a major binding pool for MeHg, while the cellular debris was an important binding pool for Hg(II). Elucidating the different subcellular fates of Hg(II) and MeHg may help us understand their toxicity in marine phytoplankton at the bottom of aquatic food chains. - Highlights: → The inter-species toxic difference of methylmercury in marine phytoplankton can be explained by its total cellular or intracellular accumulation. → The inter-species toxic difference of inorganic mercury in marine phytoplankton can be explained by its subcellular distribution. → Heat-stable protein was a major binding pool for MeHg, while the cellular debris was an important binding pool for Hg(II). - The inter-species difference in methylmercury and inorganic mercury toxicity in phytoplankton can be explained by cellular accumulation and subcellular distribution.

How uncertain is model-based prediction of copper loads in stormwater runoff?

DEFF Research Database (Denmark)

Lindblom, Erik Ulfson; Ahlman, S.; Mikkelsen, Peter Steen

2007-01-01

(runoff volumes and pollutant masses). We use the generalized likelihood uncertainty estimation (GLUE) methodology and generate posterior parameter distributions that result in model outputs encompassing a significant number of the highly variable measurements. Given the applied pollution accumulation......In this paper, we conduct a systematic analysis of the uncertainty related with estimating the total load of pollution (copper) from a separate stormwater drainage system, conditioned on a specific combination of input data, a dynamic conceptual pollutant accumulation-washout model and measurements...

Effect of Cu2+ and pH on intracellular calcium content and lipid peroxidation in winter wheat roots

Directory of Open Access Journals (Sweden)

M. E. Riazanova

2015-06-01

Full Text Available The study investigates the effect of copper ions and pH of external solution on intracellular calcium homeostasis and lipid peroxidation in winter wheat roots. Experiment was carried out with winter wheat. Sterile seeds were germinated in Petri dishes on the filter paper soaked with acetic buffer (pH 4.7 and 6.2 at 20 °Cin the dark for 48 hours. Copper was added as CuSO4. It’s concentrations varied from 0 to 50 µM. The Ca2+-fluorescent dye Fluo-3/AM ester was loaded on 60 hour. Root fluorescence with Fluo-3 loading was detected using X-Cite Series 120 Q unit attached to microscope Olympus BX53 with camera Olympus DP72. Imaging of root cells was achieved after exciting with 488 nm laser and collection of emission signals above 512 nm. Preliminary analysis of the images was performed using software LabSens; brightness (fluorescence intensity analysis was carried out by means of ImageJ. Peroxidation of lipids was determined according to Kumar and Knowles method. It was found that pH of solution had effect on release of calcium from intracellular stores. Low pH provokes an increase of [Ca2+]cyt which may be reaction of roots to acidic medium. Copper induces increase in non-selective permeability of plasma membrane and leads to its faster depolarization. This probably initiates Ca-dependent depolarization channels which are responsible for the influx of calcium from apoplast into the cell. Changing of the membrane permeability may occur due to interaction between Cu2+ ions and Ca-binding sites on plasma membrane or may be due to binding of copper with sulfhydryl groups and increasing of POL. Copper may also damage lipid bilayer and change the activity of some non-selective channels and transporters. Reactive oxygen species which are formed under some types of stress factors, especially the effect of heavy metals, can be activators of Ca-channels. Cu2+ ions rise MDA content and promote the oxidative stress. Low medium pH also induces its

Copper carrier protein in copper toxic sheep liver

Energy Technology Data Exchange (ETDEWEB)

Harris, A L; Dean, P D.G.

1973-01-01

The livers of copper-toxic sheep have been analyzed by gel electrophoresis followed by staining the gels for copper with diethyldithiocarbamate and for protein with amido schwartz. These gels were compared with similar gels obtained from the livers of normal and copper-deficient animals. The copper-toxic livers contained an extra protein band which possessed relatively weakly bound copper. Possible origins of this protein are discussed. 8 references, 1 figure, 2 tables.

Low copper and high manganese levels in prion protein plaques

Science.gov (United States)

Johnson, Christopher J.; Gilbert, P.U.P.A.; Abrecth, Mike; Baldwin, Katherine L.; Russell, Robin E.; Pedersen, Joel A.; McKenzie, Debbie

2013-01-01

Accumulation of aggregates rich in an abnormally folded form of the prion protein characterize the neurodegeneration caused by transmissible spongiform encephalopathies (TSEs). The molecular triggers of plaque formation and neurodegeneration remain unknown, but analyses of TSE-infected brain homogenates and preparations enriched for abnormal prion protein suggest that reduced levels of copper and increased levels of manganese are associated with disease. The objectives of this study were to: (1) assess copper and manganese levels in healthy and TSE-infected Syrian hamster brain homogenates; (2) determine if the distribution of these metals can be mapped in TSE-infected brain tissue using X-ray photoelectron emission microscopy (X-PEEM) with synchrotron radiation; and (3) use X-PEEM to assess the relative amounts of copper and manganese in prion plaques in situ. In agreement with studies of other TSEs and species, we found reduced brain levels of copper and increased levels of manganese associated with disease in our hamster model. We also found that the in situ levels of these metals in brainstem were sufficient to image by X-PEEM. Using immunolabeled prion plaques in directly adjacent tissue sections to identify regions to image by X-PEEM, we found a statistically significant relationship of copper-manganese dysregulation in prion plaques: copper was depleted whereas manganese was enriched. These data provide evidence for prion plaques altering local transition metal distribution in the TSE-infected central nervous system.

Investigation of heavy metal accumulation in Polygonum thunbergii for phytoextraction

Energy Technology Data Exchange (ETDEWEB)

Kim, In Sung; Kang, Kyung Hong; Johnson-Green, Perry; Lee, Eun Ju

2003-11-01

Polygonum thunbergii is useful to remove heavy metal from soil and water. - In this study, cadmium (II), lead (II), copper (II) and zinc (II) were determined in Polygonum thunbergii and soil from the Mankyung River watershed, Korea. Soil samples contained detectable lead (<17.5 {mu}g g{sup -1}), copper (<8.4 {mu}g g{sup -1}) and zinc (<24.5 {mu}g g{sup -1}), whereas cadmium was undetectable. Whole plants of P. thunbergii contained detectable lead (<320.8 {mu}g g{sup -1}), copper (<863.2 {mu}g g{sup -1}) and zinc (<2427.3 {mu}g g{sup -1}), whereas cadmium was detectable only in the stem (<7.4 {mu}g g{sup -1}) and root (<10.1 {mu}g g{sup -1}). Whole plant concentrations were very different for each metal, particularly in the case of zinc. The mean content of heavy metal in the whole plants increased in the order of cadmium (8.5 {mu}g g{sup -1})copper (548.1 {mu}g g{sup -1})copper and zinc were correlated with each metal's accumulation in the plants (lead, r=0.841, P<0.005; copper, r=0.874, P<0.001; zinc, r=0.770, P<0.005). Lead content in roots and leaves was highly correlated (r=0.5529, P<0.001), as was lead content in roots and stems (r=0.5425, P<0.001). Mean bioconcentration factors for the aboveground tissues were 4.2 (lead), 14.8 (copper) and 27.7 (zinc), and for the underground tissues, were 22.2 (lead), 92.9 (copper) and 62.7 (zinc). After hydroponic growth, bioaccumulation coefficients were 2.0 (cadmium), 3.2 (lead), 17.2 (copper) and 13.1 (zinc) for whole plants. We considered these results as indicative of the ability of P. thunbergii plants to take up metal ions from a soil matrix contaminated with heavy metals.

Horizontal Transmission of Intracellular Insect Symbionts via Plants

Directory of Open Access Journals (Sweden)

Ewa Chrostek

2017-11-01

Full Text Available Experimental evidence is accumulating that endosymbionts of phytophagous insects may transmit horizontally via plants. Intracellular symbionts known for manipulating insect reproduction and altering fitness (Rickettsia, Cardinium, Wolbachia, and bacterial parasite of the leafhopper Euscelidius variegatus have been found to travel from infected insects into plants. Other insects, either of the same or different species can acquire the symbiont from the plant through feeding, and in some cases transfer it to their progeny. These reports prompt many questions regarding how intracellular insect symbionts are delivered to plants and how they affect them. Are symbionts passively transported along the insect-plant-insect path, or do they actively participate in the process? How widespread are these interactions? How does symbiont presence influence the plant? And what conditions are required for the new infection to establish in an insect? From an ecological, evolutionary, and applied perspective, this mode of horizontal transmission could have profound implications if occurring frequently enough or if new stable symbiont infections are established. Transmission of symbionts through plants likely represents an underappreciated means of infection, both in terms of symbiont epidemiology and the movement of symbionts to new host species.

Copper corrosion and its relationship to solar collectors:a compendium.

Energy Technology Data Exchange (ETDEWEB)

Menicucci, David F.; Mahoney, Alan Roderick

2007-07-01

Copper has many fine qualities that make it a useful material. It is highly conductive of both heat and electricity, is ductile and workable, and reasonably resistant to corrosion. Because of these advantages, the solar water heating industry has been using it since the mid-1970s as the material of choice for collectors, the fundamental component of a solar water heating system. In most cases copper has performed flawlessly, but in some situations it has been known to fail. Pitting corrosion is the usual failure mode, but erosion can also occur. In 2000 Sandia National Laboratories and the Copper Development Association were asked to analyze the appearance of pin-hole leaks in solar collector units installed in a housing development in Arizona, and in 2002 Sandia analyzed a pitting corrosion event that destroyed a collector system at Camp Pendleton. This report includes copies of the reports and accounts of these corrosion failures, and provides a bibliography with references to many papers and articles that might be of benefit to the solar community. It consolidates in a single source information that has been accumulated at Sandia relative to copper corrosion, especially as it relates to solar water heaters.

The effect of hole transporting layer in charge accumulation properties of p-i-n perovskite solar cells

Directory of Open Access Journals (Sweden)

Fedros Galatopoulos

2017-07-01

Full Text Available The charge accumulation properties of p-i-n perovskite solar cells were investigated using three representative organic and inorganic hole transporting layer (HTL: (a Poly(3,4-ethylenedioxythiophene-poly(styrenesulfonate (PEDOT:PSS, Al 4083, (b copper-doped nickel oxide (Cu:NiOx, and (c Copper oxide (CuO. Through impedance spectroscopy analysis and modelling, it is shown that charge accumulation is decreased in the HTL/perovskite interface, between PEDOT:PSS to Cu:NiOx and CuO. This was indicative from the decrease in double layer capacitance (Cdl and interfacial charge accumulation capacitance (Cel, resulting in an increase to recombination resistance (Rrec, thus decreased charge recombination events between the three HTLs. Through AFM measurements, it is also shown that the reduced recombination events (followed by the increase in Rrec are also a result of increased grain size between the three HTLs, thus reduction in the grain boundary area. These charge accumulation properties of the three HTLs have resulted in an increase to the power conversion efficiency between the PEDOT:PSS (8.44%, Cu:NiOx (11.45%, and CuO (15.3%-based devices.

Photogrammetry for environmental monitoring: the use of drones and hydrological models for detection of soil contaminated by copper.

Science.gov (United States)

Capolupo, Alessandra; Pindozzi, Stefania; Okello, Collins; Fiorentino, Nunzio; Boccia, Lorenzo

2015-05-01

Campania Region of Southern Italy has a complex environmental situation, due to geogenic and anthropogenic soil pollution. Some of the pollutants such as copper are mobilized in the organic matter. It has been shown that wetlands provide physical as well as biogeochemical barriers against pollutants. Therefore, the objective of this study was to introduce and test an innovative approach able to predict copper accumulation points at plot scales, using a combination of aerial photos, taken by drones, micro-rill network modelling and wetland prediction indices usually used at catchment scales. Data were collected from an area measuring 4500 m(2) in Trentola Ducenta locality of Caserta Province of southern Italy. The photos processing with a fifth generation software for photogrammetry resulted in a high resolution Digital Elevation Model (DEM), used to study micro-rill processes. The DEM was also used to test the ability of Topographic Index (TI) and the Clima-Topographic Index (CTI) to predict copper sedimentation points at plot scale (0.1-10 ha) by comparing the map of the predicted and the actual copper distribution in the field. The DEM obtained with a resolution of 30 mm showed a high potential for the study of micro-rill processes and TI and CTI indices were able to predict zones of copper accumulation at a plot scale. Copyright © 2015 Elsevier B.V. All rights reserved.

Copper influence on bank vole's (Myodes glareolus) sexual behavior.

Science.gov (United States)

Miska-Schramm, Agata; Kapusta, Joanna; Kruczek, Małgorzata

2018-04-01

The impact of human activity on the environment has led to a steady increase of the amounts of copper in the ecosystems. This element accumulates in plants and water, potentially exposing rodents to its harmful effects. In industrial districts, a decrease in the density of small rodent populations has been observed. This decline may be caused by many factors, including mortality, decreased fertility, or impaired sexual behavior. The decline in the reproductive abilities of small rodents after copper exposure was demonstrated in our previous work (Miska-Schramm A, Kruczek M, Kapusta J, Ecotoxicology 23:1546-1554, 2014). The aim of the presented research was to determine how copper administered at concentrations similar to those recorded in industrial districts (Cu I-150 mg/kg, Cu II-600 mg/kg, C-control) affects the sexual behavior of small rodents. The model species was the bank vole (Myodes glareolus). The behavior and vocalizations of male-female pairs were recorded during open-field tests: ♂C vs. ♀C; ♂Cu I vs. ♀C; ♂Cu II vs. ♀C while in preference tests, female behavior was assessed in the following combinations: ♀C vs. ♂C & ♂Cu I; ♀C vs. ♂C & ♂Cu II. In the presented work, we show that copper decreased the males' sexual attractiveness. Females showed suppressed preference towards males treated with 600 mg/kg copper. The number of sniffs and a number of approaches towards Cu II males was significantly lower than towards control individuals. Also, in preference test with 150 mg/kg treated animals, total activity was lower towards copper treated animals. At the same time, copper did not influence intra-sexual interactions.

Copper and copper-nickel-alloys - An overview

Energy Technology Data Exchange (ETDEWEB)

Klassert, Anton; Tikana, Ladji [Deutsches Kupferinstitut e.V. Am Bonneshof 5, 40474 Duesseldorf (Germany)

2004-07-01

With the increasing level of industrialization the demand for and the number of copper alloys rose in an uninterrupted way. Today, the copper alloys take an important position amongst metallic materials due to the large variety of their technological properties and applications. Nowadays there exist over 3.000 standardized alloys. Copper takes the third place of all metals with a worldwide consumption of over 15 millions tons per year, following only to steel and aluminum. In a modern industrial society we meet copper in all ranges of the life (electro-technology, building and construction industry, mechanical engineering, automotive, chemistry, offshore, marine engineering, medical applications and others.). Copper is the first metal customized by humanity. Its name is attributed to the island Cyprus, which supplied in the antiquity copper to Greece, Rome and the other Mediterranean countries. The Romans called it 'ore from Cyprus' (aes cyprium), later cuprum. Copper deposited occasionally also dapper and could be processed in the recent stone age simply by hammering. Already in early historical time copper alloys with 20 to 50 percent tin was used for the production of mirrors because of their high reflecting power. Although the elementary nickel is an element discovered only recently from a historical perspective, its application in alloys - without any knowledge of the alloy composition - occurred at least throughout the last 2.000 years. The oldest copper-nickel coin originates from the time around 235 B.C.. Only around 1800 AD nickel was isolated as a metallic element. In particular in the sea and offshore technology copper nickel alloys found a broad field of applications in piping systems and for valves and armatures. The excellent combination of characteristics like corrosion resistance, erosion stability and bio-fouling resistance with excellent mechanical strength are at the basis of this success. An experience of many decades supports the use

Apoplastic and intracellular plant sugars regulate developmental transitions in witches' broom disease of cacao.

Science.gov (United States)

Barau, Joan; Grandis, Adriana; Carvalho, Vinicius Miessler de Andrade; Teixeira, Gleidson Silva; Zaparoli, Gustavo Henrique Alcalá; do Rio, Maria Carolina Scatolin; Rincones, Johana; Buckeridge, Marcos Silveira; Pereira, Gonçalo Amarante Guimarães

2015-03-01

Witches' broom disease (WBD) of cacao differs from other typical hemibiotrophic plant diseases by its unusually long biotrophic phase. Plant carbon sources have been proposed to regulate WBD developmental transitions; however, nothing is known about their availability at the plant-fungus interface, the apoplastic fluid of cacao. Data are provided supporting a role for the dynamics of soluble carbon in the apoplastic fluid in prompting the end of the biotrophic phase of infection. Carbon depletion and the consequent fungal sensing of starvation were identified as key signalling factors at the apoplast. MpNEP2, a fungal effector of host necrosis, was found to be up-regulated in an autophagic-like response to carbon starvation in vitro. In addition, the in vivo artificial manipulation of carbon availability in the apoplastic fluid considerably modulated both its expression and plant necrosis rate. Strikingly, infected cacao tissues accumulated intracellular hexoses, and showed stunted photosynthesis and the up-regulation of senescence markers immediately prior to the transition to the necrotrophic phase. These opposite findings of carbon depletion and accumulation in different host cell compartments are discussed within the frame of WBD development. A model is suggested to explain phase transition as a synergic outcome of fungal-related factors released upon sensing of extracellular carbon starvation, and an early senescence of infected tissues probably triggered by intracellular sugar accumulation. © The Author 2014. Published by Oxford University Press on behalf of the Society for Experimental Biology.

Chronic intoxication with copper in sheep: prophylaxis and treatment

Energy Technology Data Exchange (ETDEWEB)

Tomov, A

1964-01-01

In the period 1961-1962 chronic intoxication of sheep with copper was observed on 7 farms, the number of sheep totaling about 9000. In one of the flocks intoxication occurred after feeding the sheep for a long time with aftermath collected from orchards which had been sprinkled with copper compounds. The aftermath was proven to contain from 11 to 17 mg. per cent of copper. The other flocks were put to pasture in vineyards after the grapes had been gathered. Intoxication in these flocks was caused by vineleaves. No accumulation of copper through the soil was established in the investigated fresh grass found in the vineyards and which the sheep had also grazed. From 36 to 81 mg. per cent of copper was established in the livers of dead sheep. In order to protect the sheep from this intoxication, 100-200 mg. of ammonium or sodium molybdate and 5-10 g. of sodium sulfate were used in the case of each sheep daily for 2-3 weeks. The concentrated fodder of the sheep was moistened with an aqueous solution of these compounds. A very good prophylactic effect was obtained. At the same time oats, maize, oilcakes and others, which constitute a fodder rich in fats, were eliminated from the food rations of the sheep. The above substances, given at larger doses simultaneously with vitamin B/sub 12/ were also used for the treatment of diseased sheep, and of 10 ill sheep, 7 recovered.

Drosophila VAMP7 regulates Wingless intracellular trafficking.

Science.gov (United States)

Gao, Han; He, Fang; Lin, Xinhua; Wu, Yihui

2017-01-01

Drosophila Wingless (Wg) is a morphogen that determines cell fate during development. Previous studies have shown that endocytic pathways regulate Wg trafficking and signaling. Here, we showed that loss of vamp7, a gene required for vesicle fusion, dramatically increased Wg levels and decreased Wg signaling. Interestingly, we found that levels of Dally-like (Dlp), a glypican that can interact with Wg to suppress Wg signaling at the dorsoventral boundary of the Drosophila wing, were also increased in vamp7 mutant cells. Moreover, Wg puncta in Rab4-dependent recycling endosomes were Dlp positive. We hypothesize that VAMP7 is required for Wg intracellular trafficking and the accumulation of Wg in Rab4-dependent recycling endosomes might affect Wg signaling.

Particle Based Alloying by Accumulative Roll Bonding in the System Al-Cu

Directory of Open Access Journals (Sweden)

Mathias Göken

2011-11-01

Full Text Available The formation of alloys by particle reinforcement during accumulative roll bonding (ARB, and subsequent annealing, is introduced on the basis of the binary alloy system Al-Cu, where strength and electrical conductivity are examined in different microstructural states. An ultimate tensile strength (UTS of 430 MPa for Al with 1.4 vol.% Cu was reached after three ARB cycles, which almost equals UTS of the commercially available Al-Cu alloy AA2017A with a similar copper content. Regarding electrical conductivity, the UFG structure had no significant influence. Alloying of aluminum with copper leads to a linear decrease in conductivity of 0.78 µΩ∙cm/at.% following the Nordheim rule. On the copper-rich side, alloying with aluminum leads to a slight strengthening, but drastically reduces conductivity. A linear decrease of electrical conductivity of 1.19 µΩ∙cm/at.% was obtained.

Precursors for formation of copper selenide, indium selenide, copper indium diselenide, and/or copper indium gallium diselenide films

Science.gov (United States)

Curtis, Calvin J; Miedaner, Alexander; Van Hest, Maikel; Ginley, David S

2014-11-04

Liquid-based precursors for formation of Copper Selenide, Indium Selenide, Copper Indium Diselenide, and/or copper Indium Galium Diselenide include copper-organoselenides, particulate copper selenide suspensions, copper selenide ethylene diamine in liquid solvent, nanoparticulate indium selenide suspensions, and indium selenide ethylene diamine coordination compounds in solvent. These liquid-based precursors can be deposited in liquid form onto substrates and treated by rapid thermal processing to form crystalline copper selenide and indium selenide films.

Intracellular Na+ regulation of Na+ pump sites in cultured vascular smooth muscle cells

International Nuclear Information System (INIS)

Allen, J.C.; Navran, S.S.; Seidel, C.L.; Dennison, D.K.; Amann, J.M.; Jemelka, S.K.

1989-01-01

Enzymatically dispersed cells from canine saphenous vein and femoral artery were grown in fetal calf serum and studied at day 0 (freshly dispersed) through confluence in primary culture. Intracellular Na levels (Nai), but not intracellular K (Ki), were increased after 24 h in culture and then decreased to a steady state by 4 days. Na+ pump site number [( 3 H] ouabain binding) increased through day 3 and remained elevated. Nai was still elevated at 2 days when the Na+ pump site number began to increase. Total pump turnover (maximum ouabain-inhibited 86 Rb uptake) reflected the increase in Na+ pump site number. These key events precede the observed increases in both protein production and cellular proliferation. If the same cells are maintained in defined medium, without fetal calf serum, Nai, Ki, and the number of [ 3 H]ouabain binding sites do not change with time. These data are consistent with the suggestion that the initial mitogenic response of vascular smooth muscle cells to fetal calf serum involves an increased Na+ influx, and a Nai accumulation, caused by low Na+ pump density. The synthesis of new pump sites effects a decrease in the accumulated Nai, which may be related to cell proliferation

CCS and SOD1 mRNA are reduced after copper supplementation in peripheral mononuclear cells of individuals with high serum ceruloplasmin concentration.

Science.gov (United States)

Suazo, Miriam; Olivares, Felipe; Mendez, Marco A; Pulgar, Rodrigo; Prohaska, Joseph R; Arredondo, Miguel; Pizarro, Fernando; Olivares, Manuel; Araya, Magdalena; González, Mauricio

2008-04-01

The limits of copper homeostatic regulation in humans are not known, making it difficult to define the milder effects of early copper excess. Furthermore, a robust assay to facilitate the detection of early stages of copper excess is needed. To address these issues, we assessed changes in relative mRNA abundance of methallothionein 2A (MT2A), prion (PrP), amyloid precursor-like protein 2 (APLP2), Cu/Zn superoxide dismutase (SOD1) and its copper chaperone (CCS) in peripheral mononuclear cells (PMNCs) from healthy adults representing the 5% highest and lowest extremes in the distribution curve of serum ceruloplasmin (Cp) concentrations of 800 individuals. The intracellular Cu content was also determined. PMNCs were isolated from individuals before and after exposure to a single daily dose of 10 mg Cu (as CuSO(4)) for 2 months. Results showed that although there were fluctuations in serum Cp values of the samples assessed before copper exposure, no significant differences were observed in cell copper content or in the relative abundance of MT2A, PrP and APLP2 transcripts in PMNCs. Also, these values were not modified after copper supplementation. However, CCS and SOD1 mRNA levels were reduced in PMNCs after copper supplementation in the individuals with the high Cp values, suggesting that they should be further explored as biomarkers of moderate copper overload in humans.

Advanced Copper Composites Against Copper-Tolerant Xanthomonas perforans and Tomato Bacterial Spot.

Science.gov (United States)

Strayer-Scherer, A; Liao, Y Y; Young, M; Ritchie, L; Vallad, G E; Santra, S; Freeman, J H; Clark, D; Jones, J B; Paret, M L

2018-02-01

Bacterial spot, caused by Xanthomonas spp., is a widespread and damaging bacterial disease of tomato (Solanum lycopersicum). For disease management, growers rely on copper bactericides, which are often ineffective due to the presence of copper-tolerant Xanthomonas strains. This study evaluated the antibacterial activity of the new copper composites core-shell copper (CS-Cu), multivalent copper (MV-Cu), and fixed quaternary ammonium copper (FQ-Cu) as potential alternatives to commercially available micron-sized copper bactericides for controlling copper-tolerant Xanthomonas perforans. In vitro, metallic copper from CS-Cu and FQ-Cu at 100 μg/ml killed the copper-tolerant X. perforans strain within 1 h of exposure. In contrast, none of the micron-sized copper rates (100 to 1,000 μg/ml) from Kocide 3000 significantly reduced copper-tolerant X. perforans populations after 48 h of exposure compared with the water control (P copper-based treatments killed the copper-sensitive X. perforans strain within 1 h. Greenhouse studies demonstrated that all copper composites significantly reduced bacterial spot disease severity when compared with copper-mancozeb and water controls (P copper composites significantly reduced disease severity when compared with water controls, using 80% less metallic copper in comparison with copper-mancozeb in field studies (P copper composites have the potential to manage copper-tolerant X. perforans and tomato bacterial spot.

Paradoxical effects of density on measurement of copper tolerance in Silene paradoxa L.

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Capuana, Maurizio; Colzi, Ilaria; Buccianti, Antonella; Coppi, Andrea; Palm, Emily; Del Bubba, Massimo; Gonnelli, Cristina

2018-01-01

This work investigated if the assessment of tolerance to trace metals can depend on plant density in the experimental design. A non-metallicolous and a metallicolous populations of Silene paradoxa were hydroponically cultivated at increasing density and in both the absence (-Cu conditions) and excess of copper (+Cu conditions). In -Cu conditions, the metallicolous population showed a lower susceptibility to plant density in comparison to the non-metallicolous one, explained by a higher capacity of the metallicolous population to exploit resources. In +Cu conditions, an alleviating effect of increasing density was found in roots. Such effect was present to a greater extent in the non-metallicolous population, thus making the populations equally copper-tolerant at the highest density used. In shoots, an additive effect of increasing plant density to copper toxicity was reported. Its higher intensity in the metallicolous population reverted the copper tolerance relationship at the highest plant densities used. In both populations, a density-induced decrease in root copper accumulation was observed, thus concurring to the reported mitigation in +Cu conditions. Our work revealed the importance of density studies on the optimization of eco-toxicological bioassays and of metal tolerance assessment and it can be considered the first example of an alleviating effect of increasing plant number on copper stress in a metallophyte.

Urban Mines of Copper: Size and Potential for Recycling in the EU

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Luca Ciacci

2017-01-01

Full Text Available Copper is among the most important metals by production volume and variety of applications, providing essential materials and goods for human wellbeing. Compared to other world regions, Europe has modest natural reserves of copper and is highly dependent on imports to meet the domestic demand. Securing access to raw materials is of strategic relevance for Europe and the recycling of urban mines (also named “in-use stock” is a significant mean to provide forms of secondary copper to the European industry. A dynamic material flow analysis model is applied to characterize the flows of copper in the European Union (EU-28 from 1960 to 2014 and to determine the accumulation of this metal in the in-use stock. A scrap balance approach is applied to reconcile the flow of secondary copper sent to domestic recycling estimated through the model and that reported by historic statistics. The results show that per capita in-use stock amounts at 160–200 kg/person, and that current end-of-life recycling rate is around 60%. The quantification of historic flows provides a measure of how the European copper cycle has changed over time and how it may evolve in the future: major hindrances to recycling are highlighted and perspectives for improving the current practices at end-of-life are discussed.

Sublethal effects of contamination on the Mediterranean sponge Crambe crambe: metal accumulation and biological responses

International Nuclear Information System (INIS)

Cebrian, E.; Marti, R.; Uriz, J.M.; Turon, X.

2003-01-01

The effect of low levels of pollution on the growth, reproduction output, morphology and survival of adult sponges and settlers of the sponge Crambe crambe were examined. We transplanted sponges from a control area to a contaminated site and measured the main environmental variables (chemical and physical) of both sites during the study period. Except some punctual differences in particulate organic matter, silicates, nitrates, and water motion, most environmental variables in the water were similar at both sites during the study months. Mainly copper, lead and OM concentrations in the sediment, and water motion were significantly higher at the polluted site and may be implicated in the biological effects observed: decrease in the percentage of specimens with embryos, increase in shape irregularity and decrease in growth rate. Individuals naturally occurring at the polluted site and those transplanted there for four months accumulated ten times more copper than either untouched or transplant controls. Although lead concentration in sediment did not differ between sites, native specimens from the contaminated site accumulated this metal more than untouched controls. Vanadium concentration also tended to increase in the sponges living at or transplanted to the contaminated site but this difference was not significant. C. crambe is a reliable indicator of metal contamination since it accumulates copper, lead and vanadium in high amounts. At the contaminated site, sponge growth, fecundity and survival were inhibited, whereas sponge irregularity ending in sponge fission was promoted. All these effects may compromise the structure and dynamics of the sponge populations in sheltered, metal-contaminated habitats

Determination of sub-parts per billion levels of copper in complex matrices by adsorptive string voltammetry on a mercury electrode

International Nuclear Information System (INIS)

Shahbaazi, H.R.; Shahbaazi, H.R.; Safavi, A.; Maleki, N.

2008-01-01

The voltammetric characteristics of Cu(II)-SSA complex at the mercury electrode were investigated. An analytical method that based on the adsorptive accumulation of Cu(II)-SSA complex followed by the reduction of the complexed copper was developed for the copper determination in complex matrices in presence of the large amount of foreign ions. Adsorptive voltammetry determination showed that the Cu(II)-SSA complex behaves irreversibly exchanging two electrons on the hanging mercury drop electrode (HMDE). Factor affecting on the complextion, accumulation and stripping steps were studied and a procedure was developed. The instrumental parameters in the measurement step were also tested. The optimum conditions of pH, SSA concentration, accumulation potential and accumulation time were studied. Under optimal conditions (pH=12.9 glycin Buffer, 7 x 10 - '3 M SSA and accumulation potential -100 mV vs. Ag/ AgCl), a linear calibration graph in the range 1.25 μg L -1 to 42.5 μg L'- 1 and a detection limit of 0.8 μ L -1 were obtained. The average relative standard deviation (RSD) for seven determinations was calculated as 7 %, 5.5 % and 3 % for the concentrations between 3, 15 and 23 μg L'- 1 . The effect of foreign ions and surfactants on the peak height of Cu(II)-SSA complex was evaluated. The method was applied for the determination of the copper in different real samples such as crude oil, crude oil tank button sludge, waste water and tap water samples. The accuracy of the results was checked by ICP. (author)

Experimental study of composition and influence factors on fouling of stainless steel and copper in seawater

International Nuclear Information System (INIS)

Yang, Dazhang; Liu, Jianhua; E, Xiaoxue; Jiang, Linlin

2016-01-01

Highlights: • An increase of seawater temperature deteriorates the fouling and corrosion. • The main compositions of crystals are Mg(OH) 2 , Al(OH) 3 and their complex compounds. • The images of the seawater fouling on stainless steel and copper were shown in the paper. • A higher heat flux and A higher Reynolds number are prone to crystallization fouling accumulation in seawater. - Abstract: Metals and alloys are easily fouled in marine environment. It is a big problem for heat exchangers using cooling seawater in power plants or ships. In the paper, a seawater-fouling dynamic test device was built to investigate the composition and influence factors on fouling of stainless steel and copper in the cooling seawater system. Moreover, the static trials were performed to study the fouling and corrosion of stainless steel and copper in marine environment. The experimental results show that the seawater fouling of stainless steel is crystallization fouling, and the main elements of fouling are magnesium and aluminum. In addition, the results show that the seawater fouling of copper is corrosion fouling. In the dynamic experiments, the effects of heat flux and Reynolds number on stainless steel fouling were studied. The results show that higher heat flux and higher Reynolds number of seawater lead to the accumulation of seawater fouling.

Native copper as a natural analogue for copper canisters

International Nuclear Information System (INIS)

Marcos, N.

1989-12-01

This paper discusses the occurrence of native copper as found in geological formations as a stability analogue of copper canisters that are planned to be used for the disposal of spent nuclear fuel in the Finnish bedrock. A summary of several publications on native copper occurrences is presented. The present geochemical and geohydrological conditions in which copper is met with in its metallic state show that metallic copper is stable in a wide range of temperatures. At low temperatures native copper is found to be stable where groundwater has moderate pH (about 7), low Eh (< +100 mV), and low total dissolved solids, especially chloride. Microscopical and microanalytical studies were carried out on a dozen of rock samples containing native copper. The results reveal that the metal shows no significant alteration. Only the surface of copper grains is locally coated. In the oldest samples there exist small corrosion cracks; the age of the oldest samples is over 1,000 million years. A review of several Finnish groundwater studies suggests that there are places in Finland where the geohydrological conditions are favourable for native copper stability. (orig.)

Copper Sensing Function of Drosophila Metal-Responsive Transcription Factor-1 Is Mediated By a Tetranuclear Cu(I) Cluster

Energy Technology Data Exchange (ETDEWEB)

Chen, X.; Hua, H.; Balamurugan, K.; Kong, X.; Zhang, L.; George, G.N.; Georgiev, O.; Schaffner, W.; Giedroc, D.P.

2009-05-12

Drosophila melanogaster MTF-1 (dMTF-1) is a copper-responsive transcriptional activator that mediates resistance to Cu, as well as Zn and Cd. Here, we characterize a novel cysteine-rich domain which is crucial for sensing excess intracellular copper by dMTF-1. Transgenic flies expressing mutant dMTF-1 containing alanine substitutions of two, four or six cysteine residues within the sequence {sup 547}CNCTNCKCDQTKSCHGGDC{sup 565} are significantly or completely impaired in their ability to protect flies from copper toxicity and fail to up-regulate MtnA (metallothionein) expression in response to excess Cu. In contrast, these flies exhibit wild-type survival in response to copper deprivation thus revealing that the cysteine cluster domain is required only for sensing Cu load by dMTF-1. Parallel studies show that the isolated cysteine cluster domain is required to protect a copper-sensitive S. cerevisiae ace1 strain from copper toxicity. Cu(I) ligation by a Cys-rich domain peptide fragment drives the cooperative assembly of a polydentate [Cu{sub 4}-S{sub 6}] cage structure, characterized by a core of trigonally S{sub 3} coordinated Cu(I) ions bound by bridging thiolate ligands. While reminiscent of Cu{sub 4}-L{sub 6} (L = ligand) tetranuclear clusters in copper regulatory transcription factors of yeast, the absence of significant sequence homology is consistent with convergent evolution of a sensing strategy particularly well suited for Cu(I).

Intracellular and membrane-damaging activities of methyl gallate isolated from Terminalia chebula against multidrug-resistant Shigella spp.

Science.gov (United States)

Acharyya, Saurabh; Sarkar, Prodipta; Saha, Dhira R; Patra, Amarendra; Ramamurthy, T; Bag, Prasanta K

2015-08-01

Shigella spp. (Shigella dysenteriae, Shigella flexneri, Shigella boydii and Shigella sonnei) cause bacillary dysentery (shigellosis), which is characterized by bloody mucous diarrhoea. Although a variety of antibiotics have been effective for treatment of shigellosis, options are becoming limited due to globally emerging drug resistance. In the present study, in vitro antibacterial activity of methyl gallate (MG) isolated from Terminalia chebula was determined by performing MIC, minimal bactericidal concentration (MBC) and time-kill kinetic studies. Bacterial membrane-damaging activity of MG was determined by membrane perturbation and transmission electron microscopy (TEM). Cellular drug accumulation, cell infection and assessment of intracellular activities of MG and reference antibiotics were performed using HeLa cell cultures. The bactericidal activity of MG against multidrug-resistant (MDR) Shigella spp. in comparison with other commonly used drugs including fluoroquinolone was demonstrated here. TEM findings in the present study revealed that MG caused the total disintegration of inner and outer membranes, and leakage of the cytoplasmic contents of S. dysenteriae. The level of accumulation of MG and tetracycline in HeLa cells incubated for 24  h was relatively higher than that of ciprofloxacin and nalidixic acid (ratio of intracellular concentration/extracellular concentration of antibiotic for MG and tetracycline>ciprofloxacin and nalidixic acid). The viable number of intracellular S. dysenteriae was decreased in a time-dependent manner in the presence of MG (4 × MBC) and reduced to zero within 20  h. The significant intracellular activities of MG suggested that it could potentially be used as an effective antibacterial agent for the treatment of severe infections caused by MDR Shigella spp.

Impact of intracellular metallothionein on metal biouptake and partitioning dynamics at bacterial interfaces.

Science.gov (United States)

Présent, Romain M; Rotureau, Elise; Billard, Patrick; Pagnout, Christophe; Sohm, Bénédicte; Flayac, Justine; Gley, Renaud; Pinheiro, José P; Duval, Jérôme F L

2017-11-08

Genetically engineered microorganisms are alternatives to physicochemical methods for remediation of metal-contaminated aquifers due to their remarkable bioaccumulation capacities. The design of such biosystems would benefit from the elaboration of a sound quantitative connection between performance in terms of metal removal from aqueous solution and dynamics of the multiscale processes leading to metal biouptake. In this work, this elaboration is reported for Escherichia coli cells modified to overexpress intracellular metallothionein (MTc), a strong proteinaceous metal chelator. Depletion kinetics of Cd(ii) from bulk solution following biouptake and intracellular accumulation is addressed as a function of cell volume fraction using electroanalytical probes and ligand exchange-based analyses. It is shown that metal biouptake in the absence and presence of MTc is successfully interpreted on the basis of a formalism recently developed for metal partitioning dynamics at biointerfaces with integration of intracellular metal speciation. The analysis demonstrates how fast sequestration of metals by intracellular MTc bypasses metal excretion (efflux) and enhances the rate of metal depletion to an extent such that complete removal is achieved at sufficiently large cell volume fractions. The magnitude of the stability constant of nanoparticulate metal-MTc complexes, as derived from refined analysis of macroscopic bulk metal depletion data, is further confirmed by independent electrochemical measurement of metal binding by purified MTc extracts.

Profiling of Intracellular Metabolites: An Approach to Understanding the Characteristic Physiology of Mycobacterium leprae.

Science.gov (United States)

Miyamoto, Yuji; Mukai, Tetsu; Matsuoka, Masanori; Kai, Masanori; Maeda, Yumi; Makino, Masahiko

2016-08-01

Mycobacterium leprae is the causative agent of leprosy and also known to possess unique features such as inability to proliferate in vitro. Among the cellular components of M. leprae, various glycolipids present on the cell envelope are well characterized and some of them are identified to be pathogenic factors responsible for intracellular survival in host cells, while other intracellular metabolites, assumed to be associated with basic physiological feature, remain largely unknown. In the present study, to elucidate the comprehensive profile of intracellular metabolites, we performed the capillary electrophoresis-mass spectrometry (CE-MS) analysis on M. leprae and compared to that of M. bovis BCG. Interestingly, comparison of these two profiles showed that, in M. leprae, amino acids and their derivatives are significantly accumulated, but most of intermediates related to central carbon metabolism markedly decreased, implying that M. leprae possess unique metabolic features. The present study is the first report demonstrating the unique profiles of M. leprae metabolites and these insights might contribute to understanding undefined metabolism of M. leprae as well as pathogenic characteristics related to the manifestation of the disease.

Identification of lactoferricin B intracellular targets using an Escherichia coli proteome chip.

Science.gov (United States)

Tu, Yu-Hsuan; Ho, Yu-Hsuan; Chuang, Ying-Chih; Chen, Po-Chung; Chen, Chien-Sheng

2011-01-01

Lactoferricin B (LfcinB) is a well-known antimicrobial peptide. Several studies have indicated that it can inhibit bacteria by affecting intracellular activities, but the intracellular targets of this antimicrobial peptide have not been identified. Therefore, we used E. coli proteome chips to identify the intracellular target proteins of LfcinB in a high-throughput manner. We probed LfcinB with E. coli proteome chips and further conducted normalization and Gene Ontology (GO) analyses. The results of the GO analyses showed that the identified proteins were associated with metabolic processes. Moreover, we validated the interactions between LfcinB and chip assay-identified proteins with fluorescence polarization (FP) assays. Sixteen proteins were identified, and an E. coli interaction database (EcID) analysis revealed that the majority of the proteins that interact with these 16 proteins affected the tricarboxylic acid (TCA) cycle. Knockout assays were conducted to further validate the FP assay results. These results showed that phosphoenolpyruvate carboxylase was a target of LfcinB, indicating that one of its mechanisms of action may be associated with pyruvate metabolism. Thus, we used pyruvate assays to conduct an in vivo validation of the relationship between LfcinB and pyruvate level in E. coli. These results showed that E. coli exposed to LfcinB had abnormal pyruvate amounts, indicating that LfcinB caused an accumulation of pyruvate. In conclusion, this study successfully revealed the intracellular targets of LfcinB using an E. coli proteome chip approach.

Identification of lactoferricin B intracellular targets using an Escherichia coli proteome chip.

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Yu-Hsuan Tu

Full Text Available Lactoferricin B (LfcinB is a well-known antimicrobial peptide. Several studies have indicated that it can inhibit bacteria by affecting intracellular activities, but the intracellular targets of this antimicrobial peptide have not been identified. Therefore, we used E. coli proteome chips to identify the intracellular target proteins of LfcinB in a high-throughput manner. We probed LfcinB with E. coli proteome chips and further conducted normalization and Gene Ontology (GO analyses. The results of the GO analyses showed that the identified proteins were associated with metabolic processes. Moreover, we validated the interactions between LfcinB and chip assay-identified proteins with fluorescence polarization (FP assays. Sixteen proteins were identified, and an E. coli interaction database (EcID analysis revealed that the majority of the proteins that interact with these 16 proteins affected the tricarboxylic acid (TCA cycle. Knockout assays were conducted to further validate the FP assay results. These results showed that phosphoenolpyruvate carboxylase was a target of LfcinB, indicating that one of its mechanisms of action may be associated with pyruvate metabolism. Thus, we used pyruvate assays to conduct an in vivo validation of the relationship between LfcinB and pyruvate level in E. coli. These results showed that E. coli exposed to LfcinB had abnormal pyruvate amounts, indicating that LfcinB caused an accumulation of pyruvate. In conclusion, this study successfully revealed the intracellular targets of LfcinB using an E. coli proteome chip approach.

Formation of copper-indium-selenide and/or copper-indium-gallium-selenide films from indium selenide and copper selenide precursors

Science.gov (United States)

Curtis, Calvin J [Lakewood, CO; Miedaner, Alexander [Boulder, CO; Van Hest, Maikel [Lakewood, CO; Ginley, David S [Evergreen, CO; Nekuda, Jennifer A [Lakewood, CO

2011-11-15

Liquid-based indium selenide and copper selenide precursors, including copper-organoselenides, particulate copper selenide suspensions, copper selenide ethylene diamine in liquid solvent, nanoparticulate indium selenide suspensions, and indium selenide ethylene diamine coordination compounds in solvent, are used to form crystalline copper-indium-selenide, and/or copper indium gallium selenide films (66) on substrates (52).

The GARP Complex Is Involved in Intracellular Cholesterol Transport via Targeting NPC2 to Lysosomes.

Science.gov (United States)

Wei, Jian; Zhang, Ying-Yu; Luo, Jie; Wang, Ju-Qiong; Zhou, Yu-Xia; Miao, Hong-Hua; Shi, Xiong-Jie; Qu, Yu-Xiu; Xu, Jie; Li, Bo-Liang; Song, Bao-Liang

2017-06-27

Proper intracellular cholesterol trafficking is critical for cellular function. Two lysosome-resident proteins, NPC1 and NPC2, mediate the egress of low-density lipoprotein-derived cholesterol from lysosomes. However, other proteins involved in this process remain largely unknown. Through amphotericin B-based selection, we isolated two cholesterol transport-defective cell lines. Subsequent whole-transcriptome-sequencing analysis revealed two cell lines bearing the same mutation in the vacuolar protein sorting 53 (Vps53) gene. Depletion of VPS53 or other subunits of the Golgi-associated retrograde protein (GARP) complex impaired NPC2 sorting to lysosomes and caused cholesterol accumulation. GARP deficiency blocked the retrieval of the cation-independent mannose 6-phosphate receptor (CI-MPR) to the trans-Golgi network. Further, Vps54 mutant mice displayed reduced cellular NPC2 protein levels and increased cholesterol accumulation, underscoring the physiological role of the GARP complex in cholesterol transport. We conclude that the GARP complex contributes to intracellular cholesterol transport by targeting NPC2 to lysosomes in a CI-MPR-dependent manner. Copyright © 2017 The Author(s). Published by Elsevier Inc. All rights reserved.

COPPER AND COPPER-CONTAINING PESTICIDES: METABOLISM, TOXICITY AND OXIDATIVE STRESS

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Viktor Husak

2015-05-01

Full Text Available The purpose of this paper is to provide a brief review of the current knowledge regarding metabolism and toxicity of copper and copper-based pesticides in living organisms. Copper is an essential trace element in all living organisms (bacteria, fungi, plants, and animals, because it participates in different metabolic processes and maintain functions of organisms. The transport and metabolism of copper in living organisms is currently the subject of many studies. Copper is absorbed, transported, distributed, stored, and excreted in the body via the complex of homeostatic processes, which provide organisms with a needed constant level of this micronutrient and avoid excessive amounts. Many aspects of copper homeostasis were studied at the molecular level. Copper based-pesticides, in particularly fungicides, bacteriocides and herbicides, are widely used in agricultural practice throughout the world. Copper is an integral part of antioxidant enzymes, particularly copper-zinc superoxide dismutase (Cu,Zn-SOD, and plays prominent roles in iron homeostasis. On the other hand, excess of copper in organism has deleterious effect, because it stimulates free radical production in the cell, induces lipid peroxidation, and disturbs the total antioxidant capacity of the body. The mechanisms of copper toxicity are discussed in this review also.

Copper-resistant bacteria enhance plant growth and copper phytoextraction.

Science.gov (United States)

Yang, Renxiu; Luo, Chunling; Chen, Yahua; Wang, Guiping; Xu, Yue; Shen, Zhenguo

2013-01-01

In this study, we investigated the role of rhizospheric bacteria in solubilizing soil copper (Cu) and promoting plant growth. The Cu-resistant bacterium DGS6 was isolated from a natural Cu-contaminated soil and was identified as Pseudomonas sp. DGS6. This isolate solubilized Cu in Cu-contaminated soil and stimulated root elongation of maize and sunflower. Maize was more sensitive to inoculation with DGS6 than was sunflower and exhibited greater root elongation. In pot experiment, inoculation with DGS6 increased the shoot dry weight of maize by 49% and sunflower by 34%, and increased the root dry weight of maize by 85% and sunflower by 45%. Although the concentrations of Cu in inoculated and non-inoculated seedlings did not differ significantly, the total accumulation of Cu in the plants increased after inoculation. DGS6 showed a high ability to solubilize P and produce iron-chelating siderophores, as well as significantly improved the accumulation of P and Fe in both maize and sunflower shoots. In addition, DGS6 produced indole-3-acetic acid (IAA) and ACC deaminase, which suggests that it may modulate ethylene levels in plants. The bacterial strain DGS6 could be a good candidate for re-vegetation of Cu-contaminated sites. Supplemental materials are available for this article. Go to the publisher's online edition of International Journal of Phytoremediation to view the supplemental file.

Copper-resistant halophilic bacterium isolated from the polluted Maruit Lake, Egypt.

Science.gov (United States)

Osman, O; Tanguichi, H; Ikeda, K; Park, P; Tanabe-Hosoi, S; Nagata, S

2010-04-01

To isolate and characterize copper-resistant halophilic bacteria from the polluted Maruit Lake, Egypt and identify the role of plasmids in toxic metal resistance. We isolated strain MA2, showing high copper resistance up to the 1.5 mmol l(-1) concentration; it was also resistant to other metals such as nickel, cobalt and zinc and a group of antibiotics. Partial 16S rRNA analysis revealed that strain MA2 belonged to the genus Halomonas. Copper uptake, measured by atomic absorption spectrophotometery, was higher in the absence of NaCl than in the presence of 0.5-1.0 mol l(-1) NaCl during 5-15 min of incubation. Cell fractionation and electron microscopic observation clarified that most of the copper accumulated in the outer membrane and periplasmic fractions of the cells. Plasmid screening yielded two plasmids: pMA21 (11 kb) and pMA22 (5 kb). Plasmid curing resulted in a strain that lost both the plasmids and was sensitive to cobalt and chromate but not copper, nickel and zinc. This cured strain also showed weak growth in the presence of 0.5-1.0 mol l(-1) NaCl. Partial sequencing of both plasmids led to the identification of different toxic metals transporters but copper transporters were not identified. The highest cell viability was found in the presence of 1.0 mol l(-1) NaCl at different copper concentrations, and copper uptake was optimal in the absence of NaCl. Plasmid pMA21 encoded chromate, cobalt, zinc and cadmium transporters, whereas pMA22 encoded specific zinc and RND (resistance, nodulation, cell division) efflux transporters as well as different kinds of metabolic enzymes. Copper resistance was mainly incorporated in the chromosome. Strain MA2 is a fast and efficient tool for copper bioremediation and the isolated plasmids show significant characteristics of both toxic metal and antibiotic resistance.

Modelling the extra and intracellular uptake and discharge of heavy metals in Fontinalis antipyretica transplanted along a heavy metal and pH contamination gradient

International Nuclear Information System (INIS)

Fernandez, J.A.; Vazquez, M.D.; Lopez, J.; Carballeira, A.

2006-01-01

Samples of the aquatic bryophyte Fontinalis antipyretica Hedw. were transplanted to different sites with the aim of characterizing the kinetics of the uptake and discharge of heavy metals in the extra and intracellular compartments. The accumulation of metals in extracellular compartments, characterized by an initial rapid accumulation, then a gradual slowing down over time, fitted perfectly to a Michaelis-Menten model. The discharge of metals from the same compartment followed an inverse linear model or an inverse Michaelis-Menten model, depending on the metal. In intracellular sites both uptake and discharge occurred more slowly and progressively, following a linear model. We also observed that the acidity of the environment greatly affected metal accumulation in extracellular sites, even when the metals were present at relatively high concentrations, whereas the uptake of metals within cells was much less affected by pH. - The kinetics of uptake and discharge of heavy metals, in different cellular locations, were studied in transplanted aquatic mosses

Identification and characterization of a novel Cut family cDNA that encodes human copper transporter protein CutC

International Nuclear Information System (INIS)

Li Jixi; Ji Chaoneng; Chen Jinzhong; Yang Zhenxing; Wang Yijing; Fei, Xiangwei; Zheng Mei; Gu Xing; Wen Ge; Xie Yi; Mao Yumin

2005-01-01

Copper is an essential heavy metal trace element that plays important roles in cell physiology. The Cut family was associated with the copper homeostasis and involved in several important metabolisms, such as uptake, storage, delivery, and efflux of copper. In this study, a novel Cut family cDNA was isolated from the human fetal brain library, which encodes a 273 amino acid protein with a molecular mass of about 29.3 kDa and a calculated pI of 8.17. It was named hCutC (human copper transporter protein CutC). The ORF of hCutC gene was cloned into pQE30 vector and expressed in Escherichia coli M15. The secreted hCutC protein was purified to a homogenicity of 95% by using the Ni-NTA affinity chromatography. RT-PCR analysis showed that the hCutC gene expressed extensively in human tissues. Subcellular location analysis of hCutC-EGFP fusion protein revealed that hCutC was distributed to cytoplasm of COS-7 cells, and both cytoplasm and nucleus of AD293 cells. The results suggest that hCutC may be one shuttle protein and play important roles in intracellular copper trafficking

Disruption of the Arabidopsis CGI-58 homologue produces Chanarin–Dorfman-like lipid droplet accumulation in plants

OpenAIRE

James, Christopher N.; Horn, Patrick J.; Case, Charlene R.; Gidda, Satinder K.; Zhang, Daiyuan; Mullen, Robert T.; Dyer, John M.; Anderson, Richard G. W.; Chapman, Kent D.

2010-01-01

CGI-58 is the defective gene in the human neutral lipid storage disease called Chanarin-Dorfman syndrome. This disorder causes intracellular lipid droplets to accumulate in nonadipose tissues, such as skin and blood cells. Here, disruption of the homologous CGI-58 gene in Arabidopsis thaliana resulted in the accumulation of neutral lipid droplets in mature leaves. Mass spectroscopy of isolated lipid droplets from cgi-58 loss-of-function mutants showed they contain triacylglycerols with common...

Spatial patterns of heavy metal accumulation in sediments and macrophytes of Bellandur wetland, Bangalore.

Science.gov (United States)

Ramachandra, T V; Sudarshan, P B; Mahesh, M K; Vinay, S

2018-01-15

Heavy metals are one among the toxic chemicals and accumulation in sediments and plants has been posing serious health impacts. Wetlands aid as kidneys of the landscape and help in remediation through uptake of nutrients, heavy metals and other contaminants. The analyses of macrophytes and sediment samples help in evaluating pollution status in aquatic environment. In this study concentration of six heavy metals (Cadmium (Cd), Chromium (Cr), Copper (Cu), Nickel (Ni), Lead (Pb) and Zinc (Zn)) were assessed in sediment and dominant macrophyte samples collected from Bellandur Lake, largest Lake of Bangalore, India. Sediment samples reveal of heavy metals in the inlet regions and shore samples. The accumulation of metals in sediments were in the order of Zn > Cu > Cr > Pb > Ni > Cd. All metals exceeded the critical limits of metals in the sediment. Concentration of different metals in the macrophyte samples ranked as: Cr > Cu > Zn > Pb > Ni > Cd. Chromium and Copper were found to be more than critical range. Typha angustata had the higher accumulation of all metals except chromium. Copyright © 2017 Elsevier Ltd. All rights reserved.

Medium pH in submerged cultivation modulates differences in the intracellular protein profile of Fusarium oxysporum.

Science.gov (United States)

da Rosa-Garzon, Nathália Gonsales; Laure, Hélen Julie; Souza-Motta, Cristina Maria de; Rosa, José César; Cabral, Hamilton

2017-08-09

Fusarium oxysporum is a filamentous fungus that damages a wide range of plants and thus causes severe crop losses. In fungal pathogens, the genes and proteins involved in virulence are known to be controlled by environmental pH. Here, we report the influence of culture-medium pH (5, 6, 7, and 8) on the production of degradative enzymes involved in the pathogenesis of F. oxysporum URM 7401 and on the 2D-electrophoresis profile of intracellular proteins in this fungus. F. oxysporum URM 7401 was grown in acidic, neutral, and alkaline culture media in a submerged bioprocess. After 96 hr, the crude extract was processed to enzyme activity assays, while the intracellular proteins were obtained from mycelium and analyzed using 2D electrophoresis and mass spectrometry. We note that the diversity of secreted enzymes was changed quantitatively in different culture-medium pH. Also, the highest accumulated biomass and the intracellular protein profile of F. oxysporum URM 7401 indicate an increase in metabolism in neutral-alkaline conditions. The differential profiles of secreted enzymes and intracellular proteins under the evaluated conditions indicate that the global protein content in F. oxysporum URM 7401 is modulated by extracellular pH.

Detection of ubiquitinated huntingtin species in intracellular aggregates

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Katrin eJuenemann

2015-01-01

Full Text Available Protein conformation diseases, including polyglutamine diseases, result from the accumulation and aggregation of misfolded proteins. Huntington’s disease is one of nine diseases caused by an expanded polyglutamine repeat within the affected protein and is hallmarked by intracellular inclusion bodies composed of aggregated N-terminal huntingtin fragments and other sequestered proteins. Fluorescence microscopy and filter trap assay are conventional methods to study protein aggregates, but cannot be used to analyze the presence and levels of post-translational modifications of aggregated huntingtin such as ubiquitination. Ubiquitination of proteins can be a signal for degradation and intracellular localization, but also affects protein activity and protein-protein interactions. The function of ubiquitination relies on its mono- and polymeric isoforms attached to protein substrates. Studying the ubiquitination pattern of aggregated huntingtin fragments offers an important possibility to understand huntingtin degradation and aggregation processes within the cell. For the identification of aggregated huntingtin and its ubiquitinated species, solubilization of the cellular aggregates is mandatory. Here we describe methods to identify post-translational modifications such as ubiquitination of aggregated mutant huntingtin. This approach is specifically described for use with mammalian cell culture and is suitable to study other disease-related proteins prone to aggregate.

The influence of copper-based fungicide use in soils and aquatic sediments. Case study: Aetoliko lagoon, Western Greece

Science.gov (United States)

Avramidis, Pavlos; Barouchas, Pantelis; Dünwald, Thomas; Unkel, Ingmar

2017-04-01

In the study area, in order farmers to keep their olive trees healthy, the first measure is to keep their olive trees well-fed that is the best initial defense against diseases. Copper-based fungicides are the most common fungicides to protect olive plantations against diseases such as the olive leaf spot. Pathogens are controlled by farmers with strategically timed disease control programs rely on copper sprays to protect the foliage and fruit from infection Successful disease control depends on even distribution and good retention of the copper over all of the plant surfaces before the disease develops. Artificially added copper has the ability to accumulate in soils and aquatic sediments and can cause adverse effects on flora and fauna in its environment. For the present study soil and aquatic sediments field campaign was carried out in the Aetoliko Lagoon ecosystem which is exclusively dominated by olive orchards. It is for the first time in Greece that soil as well as aquatic sediments samples of one coherent protected aquatic ecosystem were taken and compared. To determine the influence that the usage of copper-based fungicides have on the lagoon and surrounding areas, ten (10) sediment samples from the bottom of the lagoon and twenty five (25) soil samples at the different olive orchards that are bordering the water body were taken. The samples were analyzed for total copper content (total digestion) and extractable copper (DTPA and NH4NO3). Furthermore, soil / sedimentological and geochemical analyses such as pH, grain size, total organic carbon, total nitrogen and calcium carbonate content were carried out. The results show in over 80 % of the orchard soils a critical accumulation of the total amount of copper. In some of the examined soils the value of 140 mg/kg(as set by the European Union as a limit for total copper in farmland) is exceeded by the factors of 2 to 4.5. Copper content in the aquatic sediments is generally lower and varies between 43.85 mg

Uptake and toxicity of arsenic, copper, and silicon in Azolla caroliniana and Lemna minor.

Science.gov (United States)

Rofkar, Jordan R; Dwyer, Daryl F; Bobak, Deanna M

2014-01-01

Here we report on the analysis of two aquatic plant species, Azolla caroliniana and Lemna minor, with respect to tolerance and uptake of co-occurring arsenic, copper, and silicon for use in engineered wetlands. Plants were cultured in nutrient solution that was amended with arsenic (0 or 20 microM), copper (2 or 78 microM), and silicon (0 or 1.8 mM) either singly or in combination. We hypothesized that arsenic and copper would negatively affect the uptake of metals, growth, and pigmentation and that silicon would mitigate those stresses. Tolerance was assessed by measuring growth of biomass and concentrations of chlorophyll and anthocyanins. Both plant species accumulated arsenic, copper, and silicon; L. minor generally had higher levels on a per biomass basis. Arsenic negatively impacted A. caroliniana, causing a 30% decrease in biomass production and an increase in the concentration of anthocyanin. Copper negatively impacted L. minor, causing a 60% decrease in biomass production and a 45% decrease in chlorophyll content. Silicon augmented the impact of arsenic on biomass production in A. caroliniana but mitigated the effect of copper on L. minor. Our results suggest that mixtures of plant species may be needed to maximize uptake of multiple contaminants in engineered wetlands.

Effects of copper-oxide nanoparticles, dissolved copper and ultraviolet radiation on copper bioaccumulation, photosynthesis and oxidative stress in the aquatic macrophyte Elodea nuttallii.

Science.gov (United States)

Regier, Nicole; Cosio, Claudia; von Moos, Nadia; Slaveykova, Vera I

2015-06-01

In this study, the uptake and sub-toxic effects of CuO nanoparticles (CuO-NPs), dissolved Cu(II) alone or in combination with UV radiation on the aquatic macrophyte Elodea nuttallii were studied. Emphasis was on Cu accumulation, growth, photosynthesis and the oxidative stress related enzymes peroxidase (POD) and superoxide dismutase (SOD). The results showed stronger Cu accumulation in plants exposed to 10 mg L(-1) CuO-NPs, corresponding to 1.4-2 mg L(-1) dissolved Cu(II), than to 256 μg L(-1) Cu(II). However, the ratio between the accumulated Cu and dissolved Cu in CuO treatments was lower than in Cu(II) treatments. Additional UV exposure increased accumulation in both treatments, with the effect being stronger for Cu accumulation from CuO-NPs than for dissolved Cu(II). Photosynthetic capacity was strongly reduced by UV treatment, whereas remained unaffected by Cu(II) or CuO-NP treatments. Similarly, the increase of SOD activity was more pronounced in the UV treatments. On the other hand, POD activity enhancement was strongest in the plants exposed to CuO-NPs for 24 h. Expression of the copper transporter COPT1 as revealed by RT-qPCR was inhibited by Cu(II) and CuO-NP treatment, limiting the uptake of excess Cu into the cells. Overall, the combined exposure of E. nuttallii to UV radiation with CuO-NPs or Cu(II) has a higher impact than exposure to CuO-NPs or Cu(II) alone. The results imply that heavy pollution of natural water with CuO-NPs or dissolved Cu might have stronger effects in combination with natural UV irradiation on organisms in situ. Copyright © 2015 Elsevier Ltd. All rights reserved.

DspA/E contributes to apoplastic accumulation of ROS in nonhost A. thaliana

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Alban eLaunay

2016-04-01

Full Text Available The bacterium Erwinia amylovora is responsible for the fire blight disease of Maleae, which provokes necrotic symptoms on aerial parts. The pathogenicity of this bacterium in hosts relies on its type three-secretion system (T3SS, a molecular syringe that allows the bacterium to inject effectors into the plant cell. E. amylovora-triggered disease in host plants is associated with the T3SS-dependent production of reactive oxygen species (ROS, although ROS are generally associated with resistance in other pathosystems. We showed previously that E. amylovora can multiply transiently in the nonhost plant Arabidopsis thaliana and that a T3SS-dependent production of intracellular ROS occurs during this interaction. In the present work we characterize the localization and source of hydrogen peroxide accumulation following E. amylovora infection. Transmission electron microscope (TEM analysis of infected tissues showed that hydrogen peroxide accumulation occurs in the cytosol, plastids, peroxisomes, and mitochondria as well as in the apoplast. Furthermore, TEM analysis showed that an E. amylovora dspA/E-deficient strain does not induce hydrogen peroxide accumulation in the apoplast. Consistently, a transgenic line expressing DspA/E accumulated ROS in the apoplast. The NADPH oxidase-deficient rbohD mutant showed a very strong reduction in hydrogen peroxide accumulation in response to E. amylovora inoculation. However, we did not find an increase in bacterial titers of E. amylovora in the rbohD mutant and the rbohD mutation did not suppress the toxicity of DspA/E when introgressed into a DspA/E-expressing transgenic line. Co-inoculation of E. amylovora with cycloheximide (CHX, which we found previously to suppress callose deposition and allow strong multiplication of E. amylovora in A. thaliana leaves, led to a strong reduction of apoplastic ROS accumulation but did not affect intracellular ROS. Our data strongly suggest that apoplastic ROS accumulation is

Acumulación de cobre en una comunidad vegetal afectada por contaminación minera en el valle de Puchuncaví, Chile central Copper accumulation in a plant community affected by mining contamination in Puchuncaví valley, central Chile

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ISABEL GONZÁLEZ

2008-06-01

Full Text Available Las especies hiperacumuladoras son capaces de acumular más de 1.000 mg kg-1 de metal en su biomasa aérea y son útiles en procesos de fitoextracción de metales en suelos contaminados por actividades mineras. Con el fin de identificar especies hiperacumuladoras representativas de las condiciones chilenas, se realizó una prospección dentro de la diversidad vegetal en el área afectada por las emisiones de la Fundición Ventanas (90-900 mg kg-1 de Cu total en suelos, así como en un área cercana a una pila de escorias de fundición (500-3.000 mg kg-1 de Cu total en suelos. Se determinaron las concentraciones de Cu en la biomasa aérea de las plantas. Los resultados indican que dentro de la diversidad del sitio existen al menos veintidós especies pseudometalofitas, es decir, ecotipos de especies comunes que son capaces de tolerar concentraciones de cobre en el suelo que para una planta normal serían tóxicas. Las especies fueron clasificadas según su concentración de cobre y mostraron en su mayoría media (200-600 mg kg-1 o baja (Hyperaccumulator plants species are capable of accumulating more than 1,000 mg Cu kg-1 in their shoots and are useful for metal phytoextraction in soils contaminated by mining activities. To identify the hyperaccumulator plants representative of the Chilean conditions, we carried out a survey of plant diversity in the área affected by the emissions of the Ventanas smelter (90-900 mg kg-1 of total Cu in soils and in a nearby área cióse to a smelter slug pile (500-3,000 mg kg-1 of total Cu in soils. Copper concentrations in the shoots of the studied plants were determined. Results indicate that there were at least twenty-two pseudometallophyte species, i.e., ecotypes of common species capable to tolérate concentrations of Cu in the soil that would be toxic for a normal plant. The species were classified by their copper accumulation and nearly all exhibited médium (200-600 mg kg-1 or low (< 200 mg kg-1

Screening of Phosphorus-Accumulating Fungi and Their Potential for Phosphorus Removal from Waste Streams.

Science.gov (United States)

Ye, Yulin; Gan, Jing; Hu, Bo

2015-11-01

While bacteria have been primarily studied for phosphorus (P) removal in wastewater treatment, fungi and their ability to accumulate intracellular polyphosphate are less investigated. P-accumulating fungal strains were screened from soybean plants and surrounding soil by flask cultivation with potato dextrose broth and KH2PO4 in this study. Mucor circinelloides was selected for its high efficiency in P removal efficiency and high cellular P content. Neisser staining and growth-curve analysis confirmed that M. circinelloides stored polyphosphate intracellularly by luxury phosphate uptake. The effect of culture medium compositions on P removal efficiency and cellular P content was also investigated. Monosaccharides (such as glucose and fructose) and organic nitrogen (N, such as urea, and peptone) promoted fungi growth and P accumulation. M. circinelloides also preferred organic phosphates. When glucose, urea, and phytic acid sodium salt were used as the carbon, N, and P source, respectively, the maximum utilization efficiency was 40.1% for P and 7.08% for cellular P content. In addition, the potential of M. circinelloides for P removal from waste streams was investigated. Compared with the non-inoculated control culture, inoculation with M. circinelloides improved the soluble P removal in treating wastewater centrate, screened manure, and digested manure.

Influence of conversion of penicillin G into a basic derivative on its accumulation and subcellular localization in cultured macrophages

International Nuclear Information System (INIS)

Renard, C.; Vanderhaeghe, H.J.; Claes, P.J.; Zenebergh, A.; Tulkens, P.M.

1987-01-01

beta-Lactam antibiotics do not accumulate in phagocytes, probably because of their acidic character. We therefore synthesized a basic derivative of penicillin G, namely, 14 C-labeled N-(3-dimethylamino-propyl)benzylpenicillinamide (ABP), and studied its uptake and subcellular localization in J774 macrophages compared with that of 14 C-labeled penicillin G. Whereas the intracellular concentration (Ci) of penicillin G remained lower than its extracellular concentration (Ce), ABP reached a Ci/Ce ratio of 4 to 5. Moreover, approximately 50% of intracellular ABP was found associated with lysosomes after isopycnic centrifugation of cell homogenates in isoosmotic Percoll or hyperosmotic sucrose gradients. The behavior of ABP was thus partly consistent with the model of de Duve et al., in which they described the intralysosomal accumulation of weak organic bases in lysosomes. Although ABP is microbiologically inactive, our results show that beta-lactam antibiotics can be driven into cells by appropriate modification. Further efforts therefore may be warranted in the design of active compounds or prodrugs that may prove useful in the chemotherapy of intracellular infections

Electrical conduction in composites containing copper core-copper

Indian Academy of Sciences (India)

Composites of nanometre-sized copper core-copper oxide shell with diameters in the range 6.1 to 7.3 nm dispersed in a silica gel were synthesised by a technique comprising reduction followed by oxidation of a suitably chosen precursor gel. The hot pressed gel powders mixed with nanometre-sized copper particles ...

Cellular and molecular responses of adult zebrafish after exposure to CuO nanoparticles or ionic copper.

Science.gov (United States)

Vicario-Parés, Unai; Lacave, Jose M; Reip, Paul; Cajaraville, Miren P; Orbea, Amaia

2018-01-01

Due to their antimicrobial, electrical and magnetic properties, copper nanoparticles (NPs) are suitable for a vast array of applications. Copper can be toxic to biota, making it necessary to assess the potential hazard of copper nanomaterials. Zebrafish (Danio rerio) were exposed to 10 µg Cu/L of CuO NPs of ≈100 nm (CuO-poly) or ionic copper to compare the effects provoked after 3 and 21 days of exposure and at 6 months post-exposure (mpe). At 21 days, significant copper accumulation was only detected in fish exposed to ionic copper. Exposure to both copper forms caused histopathological alterations that could reduce gill functionality, more markedly in the case of ionic copper. Nevertheless, at 6 mpe higher prevalences of gill lesions were detected in fish previously exposed to CuO-poly NPs. No relevant histological alterations were detected in liver, but the lysosomal membrane stability test showed significantly impaired general health status after exposure to both metal forms that lasted up to 6 mpe. 69 transcripts appeared regulated after 3 days of exposure to CuO-poly NPs, suggesting that NPs could produce oxidative stress and reduce metabolism and transport processes. Thirty transcripts were regulated after 21 days of exposure to ionic copper, indicating possible DNA damage. Genes of the circadian clock were identified as the key genes involved in time-dependent differences between the two copper forms. In conclusion, each copper form showed a distinct pattern of liver transcriptome regulation, but both caused gill histopathological alterations and long lasting impaired health status in adult zebrafish.

Copper, zinc, and cadmium in various fractions of soil and fungi in a Swedish forest.

Science.gov (United States)

Vinichuk, Mykhailo M

2013-01-01

Ectomycorrhizal fungi profoundly affect forest ecosystems through mediating nutrient uptake and maintaining forest food webs. The accumulation of metals in each transfer step from bulk soil to fungal sporocarps is not well known. The accumulation of three metals copper (Cu), zinc (Zn) and cadmium (Cd) in bulk soil, rhizosphere, soil-root interface, fungal mycelium and sporocarps of mycorrhizal fungi in a Swedish forest were compared. Concentrations of all three metals increased in the order: bulk soil soil-root interface (or rhizosphere) soil and sporocarps occurred against a concentration gradient. In fungal mycelium, the concentration of all three metals was about three times higher than in bulk soil, and the concentration in sporocarps was about two times higher than in mycelium. In terms of accumulation, fungi (mycelium and sporocarps) preferred Cd to Zn and Cu. Zinc concentration in sporocarps and to a lesser extent in mycelium depended on the concentration in soil, whereas, the uptake of Cu and Cd by both sporocarps and mycelium did not correlate with metal concentration in soil. Heavy metal accumulation within the fungal mycelium biomass in the top forest soil layer (0-5 cm) might account for ca. 5-9% of the total amount of Cu, 5-11% of Zn, and 16-32% of Cd. As the uptake of zinc and copper by fungi may be balanced, this implied similarities in the uptake mechanism.

Reagent conditions of the flotation of copper, copper - molybdenum and copper -zinc ores in foreing countries

International Nuclear Information System (INIS)

Nevaeva, L.M.

1983-01-01

Reagents-collectors and frothers, used abroad in reagent regimes of flotation of copper, copper-molybdenum and copper zinc ores, have been considered. Xanthogenates, aerofloats, xanthogenformiates, thionocarbamates are mainly used as reagents-collectors. Methylizobutylcarbinol and Daufros are used as reagents-frothers

Diaminobenzidine photoconversion is a suitable tool for tracking the intracellular location of fluorescently labelled nanoparticles at transmission electron microscopy

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M. Malatesta

2012-04-01

Full Text Available Chitosan-based nanoparticles (NPs deserve particular attention as suitable drug carriers in the field of pharmaceutics, since they are able to protect the encapsulated drugs and/or improve their efficacy by making them able to cross biological barriers (such as the blood-brain barrier and reach their intracellular target sites. Understanding the intracellular location of NPs is crucial for designing drug delivery strategies. In this study, fluorescently-labelled chitosan NPs were administered in vitro to a neuronal cell line, and diaminobenzidine (DAB photoconversion was applied to correlate fluorescence and transmission electron microscopy to precisely describe the NPs intracellular fate. This technique allowed to demonstrate that chitosan NPs easily enter neuronal cells, predominantly by endocytosis; they were found both inside membrane-bounded vesicles and free in the cytosol, and were observed to accumulate around the cell nucleus.

Comparative study of radiation damage accumulation in Cu and Fe

International Nuclear Information System (INIS)

Caturla, M.J.; Soneda, N.; Alonso, E.; Wirth, B.D.; Diaz de la Rubia, T.; Perlado, J.M.

2000-01-01

Bcc and fcc metals exhibit significant differences in behavior when exposed to neutron or heavy ion irradiation. Transmission electron microscopy (TEM) observations reveal that damage in the form of stacking fault tetrahedra (SFT) is visible in copper irradiated to very low doses, but that no damage is visible in iron irradiated to the same total dose. In order to understand and quantify this difference in behavior, we have simulated damage production and accumulation in fcc Cu and bcc Fe. We use 20 keV primary knock-on atoms (PKAs) at a homologous temperature of 0.25 of the melting point. The primary damage state was calculated using molecular dynamics (MD) with empirical, embedded-atom interatomic potentials. Damage accumulation was modeled using a kinetic Monte Carlo (kMC) algorithm to follow the evolution of all defects produced in the cascades. The diffusivities and binding energies of defects are input data for this simulation and were either extracted from experiments, the literature, or calculated using MD. MD simulations reveal that vacancy clusters are produced within the cascade core in the case of copper. In iron, most of the vacancies do not cluster during cooling of the cascade core and are available for diffusion. In addition, self-interstitial atom (SIA) clusters are produced in copper cascades but those observed in iron are smaller in number and size. The combined MD/kMC simulations reveal that the visible cluster densities obtained as a function of dose are at least one order of magnitude lower in Fe than in Cu. We compare the results with experimental measurements of cluster density and find excellent agreement between the simulations and experiments when small interstitial clusters are considered to be mobile as suggested by recent MD simulations

Xylella fastidiosa differentially accumulates mineral elements in biofilm and planktonic cells.

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Cobine, Paul A; Cruz, Luisa F; Navarrete, Fernando; Duncan, Daniel; Tygart, Melissa; De La Fuente, Leonardo

2013-01-01

Xylella fastidiosa is a bacterial plant pathogen that infects numerous plant hosts. Disease develops when the bacterium colonizes the xylem vessels and forms a biofilm. Inductively coupled plasma optical emission spectroscopy was used to examine the mineral element content of this pathogen in biofilm and planktonic states. Significant accumulations of copper (30-fold), manganese (6-fold), zinc (5-fold), calcium (2-fold) and potassium (2-fold) in the biofilm compared to planktonic cells were observed. Other mineral elements such as sodium, magnesium and iron did not significantly differ between biofilm and planktonic cells. The distribution of mineral elements in the planktonic cells loosely mirrors the media composition; however the unique mineral element distribution in biofilm suggests specific mechanisms of accumulation from the media. A cell-to-surface attachment assay shows that addition of 50 to 100 µM Cu to standard X. fastidiosa media increases biofilm, while higher concentrations (>200 µM) slow cell growth and prevent biofilm formation. Moreover cell-to-surface attachment was blocked by specific chelation of copper. Growth of X. fastidiosa in microfluidic chambers under flow conditions showed that addition of 50 µM Cu to the media accelerated attachment and aggregation, while 400 µM prevented this process. Supplementation of standard media with Mn showed increased biofilm formation and cell-to-cell attachment. In contrast, while the biofilm accumulated Zn, supplementation to the media with this element caused inhibited growth of planktonic cells and impaired biofilm formation. Collectively these data suggest roles for these minerals in attachment and biofilm formation and therefore the virulence of this pathogen.

Combined toxicity of microcystin-LR and copper on lettuce (Lactuca sativa L.).

Science.gov (United States)

Cao, Qing; Steinman, Alan D; Wan, Xiang; Xie, Liqiang

2018-05-10

Microcystins and copper commonly co-exist in the natural environment, but their combined toxicity remains unclear, especially in terrestrial plants. The present study investigated the toxicity effects of microcystin-LR (0, 5, 50, 500, 1000 μg L -1 ) and copper (0, 50, 500, 1000, 2000 μg L -1 ), both individually and in mixture, on the germination, growth and oxidative response of lettuce. The bioaccumulation of microcystin-LR and copper was also evaluated. Results showed that the decrease in lettuce germination induced by copper alone was not significantly different from that induced by the mixture, and the combined toxicity assessment showed a simple additive effect. Lettuce growth was not significantly reduced by microcystin-LR alone, whereas it was significantly reduced by copper alone and the mixture when copper concentration was higher than 500 μg L -1 . High concentrations of microcystin-LR (1000 μg L -1 ) and copper (≥50 μg L -1 ),as well as their mixture (≥50 + 500 μg L -1 ), induced oxidative stress in lettuce. A synergistic effect on the growth and antioxidative system of lettuce was observed when exposed to low concentrations of the mixture (≤50 + 500 μg L -1 ), whereas an antagonistic effect was observed at high concentrations (≥1000 + 2000 μg L -1 ). Moreover, the interaction of microcystin-LR and copper can increase their accumulation in lettuce. Our results suggest that the toxicity effects of microcystin-LR and copper are exacerbated when they co-exist in the natural environment at low concentrations, which not only negatively affects plant growth but also poses a potential risk to human health via the food chain. Copyright © 2018. Published by Elsevier Ltd.

Control of biofouling on titanium condenser tubes with the use of electroless copper plating

International Nuclear Information System (INIS)

Anandkumar, B.; George, R.P.; Kamachi Mudali, U.; Ramachandran, D.

2015-01-01

In sea water environments titanium condenser tubes face serious issues of biofouling and biomineralization. Electroless plating of nanocopper film is attempted inside the tubes for the control of biofilm formation. Using advanced techniques like AFM, SEM, and XPS, electroless copper plated flat Ti specimens were characterized. Examination of Cu coated Ti surfaces using AFM and SEM showed more reduction in the microroughness compared to anodized Ti surface. Cu 2p 3/2 peak in XPS spectral analysis showed the shift in binding energy inferring the reduction of the hydroxide to metallic copper. Tubular specimens were exposed to sea water up to three months and withdrawn at monthly intervals to evaluate antibacterial activity and long term stability of the coating. Total viable counts and epifluorescence microscopy analyses showed two orders decrease in bacterial counts on copper coated Ti specimens when compared to as polished control Ti specimens. Molecular biology techniques like DGGE and protein expression analysis system were done to get insight into the community diversity and copper tolerance of microorganisms. DGGE gel bands clearly showed the difference in the bacterial diversity inferring from the 16S rRNA gene fragments (V3 regions). Protein analysis showed distinct protein spots appearing in electroless copper coated Ti biofilm protein samples in addition to protein spots common to both the biofilms of Cu coated and as polished Ti. The results indicated copper accumulating proteins in copper resistant bacterial species of biofilm. Reduced microroughness of the surface and toxic copper ions resulted in good biofouling control even after three months exposure to sea water. (author)

Cover crops influence soil microorganisms and phytoextraction of copper from a moderately contaminated vineyard.

Science.gov (United States)

Mackie, K A; Schmidt, H P; Müller, T; Kandeler, E

2014-12-01

We investigated the ability of summer (Avena sativa [oat], Trifolium incarnatum [crimson clover], Chenopodium [goosefoot]) and winter (Vicia villosa [hairy vetch], Secale Cereale L. [Rye], Brassica napus L. partim [rape]) cover crops, including a mixed species treatment, to extract copper from an organic vineyard soil in situ and the microbial communities that may support it. Clover had the highest copper content (14.3mgCukg(-1) DM). However, it was the amount of total biomass production that determined which species was most effective at overall copper removal per hectare. The winter crop rye produced significantly higher amounts of biomass (3532kgDMha(-1)) and, therefore, removed significantly higher amounts of copper (14,920mgCuha(-1)), despite less accumulation of copper in plant shoots. The maximum annual removal rate, a summation of best performing summer and winter crops, would be 0.033kgCuha(-1)y(-1). Due to this low annual extraction efficiency, which is less than the 6kgCuha(-1)y(-1) permitted for application, phytoextraction cannot be recommended as a general method of copper extraction from vineyards. Copper concentration did not influence aboveground or belowground properties, as indicated by sampling at two distances from the grapevine row with different soil copper concentrations. Soil microorganisms may have become tolerant to the copper levels at this site. Microbial biomass and soil enzyme activities (arylsulfatase and phosphatase) were instead driven by seasonal fluxes of resource pools. Gram+ bacteria were associated with high soil moisture, while fungi seemed to be driven by extractable carbon, which was linked to high plant biomass. There was no microbial group associated with the increased phytoextraction of copper. Moreover, treatment did not influence the abundance, activity or community structure of soil microorganisms. Copyright © 2014 Elsevier B.V. All rights reserved.

Improving Beneficiation of Copper and Iron from Copper Slag by Modifying the Molten Copper Slag

Directory of Open Access Journals (Sweden)

Zhengqi Guo

2016-04-01

Full Text Available In the paper, a new technology was developed to improve the beneficiation of copper and iron components from copper slag, by modifying the molten slag to promote the mineralization of valuable minerals and to induce the growth of mineral grains. Various parameters, including binary basicity, dosage of compound additive, modification temperature, cooling rate and the end point temperature of slow cooling were investigated. Meanwhile, optical microscope, scanning electron microscope and energy dispersive spectrometer (SEM-EDS was employed to determine the mineralogy of the modified and unmodified slag, as well as to reveal the mechanisms of enhancing beneficiation. The results show that under the proper conditions, the copper grade of rougher copper concentrate was increased from 6.43% to 11.04%, iron recovery of magnetic separation was increased significantly from 32.40% to 63.26%, and other evaluation indexes were changed slightly, in comparison with unmodified copper slag. Moreover, matte and magnetite grains in the modified slag aggregated together and grew obviously to the mean size of over 50 μm, resulting in an improvement of beneficiation of copper and iron.

21 CFR 73.1125 - Potassium sodium copper chloropyhllin (chlorophyllin-copper complex).

Science.gov (United States)

2010-04-01

... 21 Food and Drugs 1 2010-04-01 2010-04-01 false Potassium sodium copper chloropyhllin (chlorophyllin-copper complex). 73.1125 Section 73.1125 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT....1125 Potassium sodium copper chloropyhllin (chlorophyllin-copper complex). (a) Identity. (1) The color...

21 CFR 73.2125 - Potassium sodium copper chlorophyllin (chlorophyllin-copper complex).

Science.gov (United States)

2010-04-01

... 21 Food and Drugs 1 2010-04-01 2010-04-01 false Potassium sodium copper chlorophyllin (chlorophyllin-copper complex). 73.2125 Section 73.2125 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT... § 73.2125 Potassium sodium copper chlorophyllin (chlorophyllin-copper complex). (a) Identity and...

Intracellular sphingosine releases calcium from lysosomes.

Science.gov (United States)

Höglinger, Doris; Haberkant, Per; Aguilera-Romero, Auxiliadora; Riezman, Howard; Porter, Forbes D; Platt, Frances M; Galione, Antony; Schultz, Carsten

2015-11-27

To elucidate new functions of sphingosine (Sph), we demonstrate that the spontaneous elevation of intracellular Sph levels via caged Sph leads to a significant and transient calcium release from acidic stores that is independent of sphingosine 1-phosphate, extracellular and ER calcium levels. This photo-induced Sph-driven calcium release requires the two-pore channel 1 (TPC1) residing on endosomes and lysosomes. Further, uncaging of Sph leads to the translocation of the autophagy-relevant transcription factor EB (TFEB) to the nucleus specifically after lysosomal calcium release. We confirm that Sph accumulates in late endosomes and lysosomes of cells derived from Niemann-Pick disease type C (NPC) patients and demonstrate a greatly reduced calcium release upon Sph uncaging. We conclude that sphingosine is a positive regulator of calcium release from acidic stores and that understanding the interplay between Sph homeostasis, calcium signaling and autophagy will be crucial in developing new therapies for lipid storage disorders such as NPC.

Heavy metal accumulation in Pelargonium hortorum: Effects on growth and development

OpenAIRE

Orroño, DI; Lavado, RS

2009-01-01

Ornamental plants have been proposed for growing in heavy metal (HM) contaminated soils, and also for phytoremediation. We evaluated (1) biomass production and (2) HM accumulation in Pelargonium hortorum. Plants were grown for 16 weeks on HM (cadmium, chromium, copper, lead, nickel and zinc) enriched soils. Treatments were i) control, non-enriched soil, ii) medium concentration treatment, and iii) high concentration treatment. Four destructive harvests were carried out, and roots, stems, leav...

Intracellular accumulation of amyloid-beta - a predictor for synaptic dysfunction and neuron loss in Alzheimer's disease

Directory of Open Access Journals (Sweden)

Thomas A Bayer

2010-03-01

Full Text Available Despite of long-standing evidence that beta-amyloid (Aβ peptides have detrimental effects on synaptic function, the relationship between Aβ, synaptic and neuron loss is largely unclear. During the last years there is growing evidence that early intraneuronal accumulation of Aβ peptides is one of the key events leading to synaptic and neuronal dysfunction. Many studies have been carried out using transgenic mouse models of Alzheimer’s disease (AD which have been proven to be valuable model system in modern AD research. The present review discusses the impact of intraneuronal Aβ accumulation on synaptic impairment and neuron loss and provides an overview of currently available AD mouse models showing these pathological alterations.

Propionate induces cell swelling and K+ accumulation in shark rectal gland

International Nuclear Information System (INIS)

Feldman, G.M.; Ziyadeh, F.N.; Mills, J.W.; Booz, G.W.; Kleinzeller, A.

1989-01-01

Small organic anions have been reported to induce cell solute accumulation and swelling. To investigate the mechanism of swelling, we utilized preparations of rectal gland cells from Squalus acanthias incubated in medium containing propionate. Propionate causes cells to swell by diffusing across membranes in its nonionic form, acidifying cell contents, and activating the Na+-H+ antiporter. The Na+-H+ exchange process tends to correct intracellular pH (pHi), and thus it maintains a favorable gradient for propionic acid diffusion and allows propionate to accumulate. Activation of the Na+-H+ antiport also facilitates Na+ entry into the cell and Nai accumulation. At the same time Na+-K+-ATPase activity, unaffected by propionate, replaces Nai with Ki, whereas the K+ leak rate, decreased by propionate, allows Ki to accumulate. As judged by 86 Rb+ efflux, the reduction in K+ leak was not due to propionate-induced cell acidification or reduction in Cli concentration. Despite inducing cell swelling, propionate did not disrupt cell structural elements and F actin distribution along cell membranes

Propionate induces cell swelling and K+ accumulation in shark rectal gland

Energy Technology Data Exchange (ETDEWEB)

Feldman, G.M.; Ziyadeh, F.N.; Mills, J.W.; Booz, G.W.; Kleinzeller, A. (Mount Desert Island Biological Laboratory, Salsbury Cove, ME (USA))

1989-08-01

Small organic anions have been reported to induce cell solute accumulation and swelling. To investigate the mechanism of swelling, we utilized preparations of rectal gland cells from Squalus acanthias incubated in medium containing propionate. Propionate causes cells to swell by diffusing across membranes in its nonionic form, acidifying cell contents, and activating the Na+-H+ antiporter. The Na+-H+ exchange process tends to correct intracellular pH (pHi), and thus it maintains a favorable gradient for propionic acid diffusion and allows propionate to accumulate. Activation of the Na+-H+ antiport also facilitates Na+ entry into the cell and Nai accumulation. At the same time Na+-K+-ATPase activity, unaffected by propionate, replaces Nai with Ki, whereas the K+ leak rate, decreased by propionate, allows Ki to accumulate. As judged by {sup 86}Rb+ efflux, the reduction in K+ leak was not due to propionate-induced cell acidification or reduction in Cli concentration. Despite inducing cell swelling, propionate did not disrupt cell structural elements and F actin distribution along cell membranes.

Activation of Cu,Zn-superoxide dismutase in the absence of oxygen and the copper chaperone CCS.

Science.gov (United States)

Leitch, Jeffry M; Jensen, Laran T; Bouldin, Samantha D; Outten, Caryn E; Hart, P John; Culotta, Valeria C

2009-08-14

Eukaryotic Cu,Zn-superoxide dismutases (SOD1s) are generally thought to acquire the essential copper cofactor and intramolecular disulfide bond through the action of the CCS copper chaperone. However, several metazoan SOD1s have been shown to acquire activity in vivo in the absence of CCS, and the Cu,Zn-SOD from Caenorhabditis elegans has evolved complete independence from CCS. To investigate SOD1 activation in the absence of CCS, we compared and contrasted the CCS-independent activation of C. elegans and human SOD1 to the strict CCS-dependent activation of Saccharomyces cerevisiae SOD1. Using a yeast expression system, both pathways were seen to acquire copper derived from cell surface transporters and compete for the same intracellular pool of copper. Like CCS, CCS-independent activation occurs rapidly with a preexisting pool of apo-SOD1 without the need for new protein synthesis. The two pathways, however, strongly diverge when assayed for the SOD1 disulfide. SOD1 molecules that are activated without CCS exhibit disulfide oxidation in vivo without oxygen and under copper-depleted conditions. The strict requirement for copper, oxygen, and CCS in disulfide bond oxidation appears exclusive to yeast SOD1, and we find that a unique proline at position 144 in yeast SOD1 is responsible for this disulfide effect. CCS-dependent and -independent pathways also exhibit differential requirements for molecular oxygen. CCS activation of SOD1 requires oxygen, whereas the CCS-independent pathway is able to activate SOD1s even under anaerobic conditions. In this manner, Cu,Zn-SOD from metazoans may retain activity over a wide range of physiological oxygen tensions.

Nickel, copper and cobalt coalescence in copper cliff converter slag

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Wolf A.

2016-01-01

Full Text Available The aim of this investigation is to assess the effect of various additives on coalescence of nickel, copper and cobalt from slags generated during nickel extraction. The analyzed fluxes were silica and lime while examined reductants were pig iron, ferrosilicon and copper-silicon compound. Slag was settled at the different holding temperatures for various times in conditions that simulated the industrial environment. The newly formed matte and slag were characterized by their chemical composition and morphology. Silica flux generated higher partition coefficients for nickel and copper than the addition of lime. Additives used as reducing agents had higher valuable metal recovery rates and corresponding partition coefficients than fluxes. Microstructural studies showed that slag formed after adding reductants consisted of primarily fayalite, with some minute traces of magnetite as the secondary phase. Addition of 5 wt% of pig iron, ferrosilicon and copper-silicon alloys favored the formation of a metallized matte which increased Cu, Ni and Co recoveries. Addition of copper-silicon alloys with low silicon content was efficient in copper recovery but coalescence of the other metals was low. Slag treated with the ferrosilicon facilitated the highest cobalt recovery while copper-silicon alloys with silicon content above 10 wt% resulted in high coalescence of nickel and copper, 87 % and 72 % respectively.

Comparative Salt Stress Study on Intracellular Ion Concentration in Marine and Salt-adapted Freshwater Strains of Microalgae

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Ahmad Farhad TALEBI

2013-08-01

Full Text Available Salinity imposes significant stresses in various living organisms including microalgae. High extracellular concentration of Na+ directly influences ionic balance inside the cell and subsequently the cellular activities. In the present study, the effect of such stress on growth and intracellular ions concentration (IIC of Dunaliella salina and Chlorella Spp. was investigated. IIC was analyzed using Ion chromatography technique. D. salina showed the highest degree of resistance to increase in salinity as little changes occurred both in IIC and in growth parameters. D. salina could maintain the balance of K+ inside the cell and eject the excess Na+ even at NaCl concentrations above 1M. Moreover, D. salina accumulated β-carotene in order to protect its photosynthetic apparatus. Among Chlorella species, C. vulgaris showed signs of adaptation to high content of salinity, though it is a fresh water species by nature. Moreover, the response shown by C. vulgaris to rise in salinity was even stronger than that of C. salina, which is presumably a salt-water resistant species. In fact, C. vulgaris could maintain intracellular K+ better than C. salina in response to increasing salinity, and as a result, it could survive at NaCl concentrations as high as 0.75 M. Marine strains such as D. salina well cope with the fluctuations in salinity through the existing adaptation mechanisms i.e. maintaining the K+/N+ balance inside the cell, K+ accumulation and Na+ ejection, accumulation of photosynthetic pigments like β-carotene.

Cytoplasmic Copper Detoxification in Salmonella Can Contribute to SodC Metalation but Is Dispensable during Systemic Infection.

Science.gov (United States)

Fenlon, Luke A; Slauch, James M

2017-12-15

immune cells, a location in which most bacteria are killed. Our work examines how one particular metal, copper, is acquired by Salmonella to activate a protein important for survival within immune cells. At high levels, copper itself can inhibit Salmonella Using a strain of Salmonella that cannot detoxify intracellular copper, we also addressed the in vivo role of copper as an antimicrobial agent. Copyright © 2017 American Society for Microbiology.

Antwerp Copper Plates

DEFF Research Database (Denmark)

Wadum, Jørgen

1999-01-01

In addition to presenting a short history of copper paintings, topics detail artists’ materials and techniques, as well as aspects of the copper industry, including mining, preparation and trade routes.......In addition to presenting a short history of copper paintings, topics detail artists’ materials and techniques, as well as aspects of the copper industry, including mining, preparation and trade routes....

Characterization of Timed Changes in Hepatic Copper Concentrations, Methionine Metabolism, Gene Expression, and Global DNA Methylation in the Jackson Toxic Milk Mouse Model of Wilson Disease

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Anh Le

2014-05-01

Full Text Available Background: Wilson disease (WD is characterized by hepatic copper accumulation with progressive liver damage to cirrhosis. This study aimed to characterize the toxic milk mouse from The Jackson Laboratory (Bar Harbor, ME, USA (tx-j mouse model of WD according to changes over time in hepatic copper concentrations, methionine metabolism, global DNA methylation, and gene expression from gestational day 17 (fetal to adulthood (28 weeks. Methods: Included liver histology and relevant biochemical analyses including hepatic copper quantification, S-adenosylmethionine (SAM and S-adenosylhomocysteine (SAH liver levels, qPCR for transcript levels of genes relevant to methionine metabolism and liver damage, and DNA dot blot for global DNA methylation. Results: Hepatic copper was lower in tx-j fetuses but higher in weanling (three weeks and adult tx-j mice compared to controls. S-adenosylhomocysteinase transcript levels were significantly lower at all time points, except at three weeks, correlating negatively with copper levels and with consequent changes in the SAM:SAH methylation ratio and global DNA methylation. Conclusion: Compared to controls, methionine metabolism including S-adenosylhomocysteinase gene expression is persistently different in the tx-j mice with consequent alterations in global DNA methylation in more advanced stages of liver disease. The inhibitory effect of copper accumulation on S-adenosylhomocysteinase expression is associated with progressively abnormal methionine metabolism and decreased methylation capacity and DNA global methylation.

Effect of minerals on accumulation of Cs by fungus Saccaromyces cerevisiae.

Science.gov (United States)

Ohnuki, Toshihiko; Sakamoto, Fuminori; Yamasaki, Shinya; Kozai, Naofumi; Shiotsu, Hiroyuki; Utsunomiya, Satoshi; Watanabe, Naoko; Kozaki, Tamotsu

2015-06-01

The accumulation of Cs by unicellular fungus of Saccharomyces cerevisiae in the presence of minerals has been studied to elucidate the role of microorganisms in the migration of radioactive Cs in the environment. Two different types of experiments were employed: experiments using stable Cs to examine the effect of a carbon source on the accumulation of Cs, and accumulation experiments of radioactive Cs from agar medium containing (137)Cs and zeolite, vermiculite, phlogopite, smectite, mica, or illite as mineral supplements. In the former type of experiments, the Cs-accumulated cells were analyzed by scanning electron microscopy equipped with energy dispersive X-ray analysis (SEM-EDS). In the latter type, the radioactivity in the yeast cells was measured by an autoradiography technique. When a carbon source was present, higher amounts of Cs accumulated in the cells than in the resting condition without a carbon source. Analyses with SEM-EDS showed that no mineral formed on the cell surface. These results indicate that the yeast cells accumulate Cs by adsorption on the cell surface and intracellular accumulation. In the presence of minerals in the agar medium, the radioactivity in the yeast cells was in the order of mica > smectite, illite > vermiculite, phlogopite, zeolite. This order is inversely correlated to the ratio of the concentration of radioactive Cs between the minerals and the medium solution. These results strongly suggest that the yeast accumulates radioactive Cs competitively with minerals. Copyright © 2015 Elsevier Ltd. All rights reserved.

The metal chaperone Atox1 regulates the activity of the human copper transporter ATP7B by modulating domain dynamics.

Science.gov (United States)

Yu, Corey H; Yang, Nan; Bothe, Jameson; Tonelli, Marco; Nokhrin, Sergiy; Dolgova, Natalia V; Braiterman, Lelita; Lutsenko, Svetlana; Dmitriev, Oleg Y

2017-11-03

The human transporter ATP7B delivers copper to the biosynthetic pathways and maintains copper homeostasis in the liver. Mutations in ATP7B cause the potentially fatal hepatoneurological disorder Wilson disease. The activity and intracellular localization of ATP7B are regulated by copper, but the molecular mechanism of this regulation is largely unknown. We show that the copper chaperone Atox1, which delivers copper to ATP7B, and the group of the first three metal-binding domains (MBD1-3) are central to the activity regulation of ATP7B. Atox1-Cu binding to ATP7B changes domain dynamics and interactions within the MBD1-3 group and activates ATP hydrolysis. To understand the mechanism linking Atox1-MBD interactions and enzyme activity, we have determined the MBD1-3 conformational space using small angle X-ray scattering and identified changes in MBD dynamics caused by apo -Atox1 and Atox1-Cu by solution NMR. The results show that copper transfer from Atox1 decreases domain interactions within the MBD1-3 group and increases the mobility of the individual domains. The N-terminal segment of MBD1-3 was found to interact with the nucleotide-binding domain of ATP7B, thus physically coupling the domains involved in copper binding and those involved in ATP hydrolysis. Taken together, the data suggest a regulatory mechanism in which Atox1-mediated copper transfer activates ATP7B by releasing inhibitory constraints through increased freedom of MBD1-3 motions. © 2017 by The American Society for Biochemistry and Molecular Biology, Inc.

Speciation and leachability of copper in mine tailings from porphyry copper mining

DEFF Research Database (Denmark)

Hansen, Henrik K.; Yianatos, Juan B; Ottosen, Lisbeth M.

2005-01-01

Mine tailing from the El Teniente-Codelco copper mine situated in VI Region of Chile was analysed in order to evaluate the mobility and speciation of copper in the solid material. Mine tailing was sampled after the rougher flotation circuits, and the copper content was measured to 1150mgkg^-^1 dry...... matter. This tailing was segmented into fractions of different size intervals: 0-38, 38-45, 45-53, 53-75, 75-106, 106-150, 150-212, and >212@mm, respectively. Copper content determination, sequential chemical extraction, and desorption experiments were carried out for each size interval in order...... to evaluate the speciation of copper. It was found that the particles of smallest size contained 50-60% weak acid leachable copper, whereas only 32% of the copper found in largest particles could be leached in weak acid. Copper oxides and carbonates were the dominating species in the smaller particles...

Tribological properties of copper-based composites with copper coated NbSe2 and CNT

International Nuclear Information System (INIS)

Chen, Beibei; Yang, Jin; Zhang, Qing; Huang, Hong; Li, Hongping; Tang, Hua; Li, Changsheng

2015-01-01

Graphical abstract: Morphology of copper coated NbSe 2 and CNT; friction coefficient and wear rate of copper-based composites. - Highlights: • NbSe 2 and CNT were coated with copper layers by the means of electroless plating. • The mechanical and tribological properties of copper composites were studied. • The enhancement mechanisms of copper coated NbSe 2 and CNT were proposed. • Copper–copper coated (12 wt.%NbSe 2 –3 wt.%CNT) composite had the best wear resistance. - Abstract: Copper-based composites with copper coated NbSe 2 and/or CNT were fabricated by the powder metallurgy technique. The morphology and phase composition of copper coated NbSe 2 and carbon nanotube (CNT) were observed using high solution transmission electronic microscope (HRTEM), scanning electronic microscope (SEM equipped with EDS) and X-ray diffraction (XRD). The density, hardness, and bending strength of as-prepared copper-based composites were measured, and their tribological properties were investigated using UMT-2 tester. Results indicated that all copper-based composites showed decreased density and bending strength, but increased hardness in comparison with copper matrix. Besides, the incorporation of copper coated NbSe 2 improved the friction-reducing and anti-wear properties of copper matrix. Addition of copper coated CNT greatly enhanced the mechanical and tribological properties. In particular, when the content of copper coated CNT was 3 wt.%, the corresponding composite exhibited the best tribological properties. This was because NbSe 2 was distributed chaotically in matrix, which greatly improved the friction-reducing property of copper, while CNT with superior mechanical strength enhanced the wear resistance by increasing the load-carrying capacity. More importantly, copper layers coated on NbSe 2 and CNT favored the good interfacial combination between fillers and copper matrix showing beneficial effect for the stresses transferring from matrix to fillers

Copper and Anesthesia: Clinical Relevance and Management of Copper Related Disorders

OpenAIRE

Langley, Adrian; Dameron, Charles T.

2013-01-01

Recent research has implicated abnormal copper homeostasis in the underlying pathophysiology of several clinically important disorders, some of which may be encountered by the anesthetist in daily clinical practice. The purpose of this narrative review is to summarize the physiology and pharmacology of copper, the clinical implications of abnormal copper metabolism, and the subsequent influence of altered copper homeostasis on anesthetic management.

Effects of chronic exposure to waterborne copper and nickel in binary mixture on tissue-specific metal accumulation and reproduction in fathead minnow (Pimephales promelas).

Science.gov (United States)

Driessnack, Melissa K; Jamwal, Ankur; Niyogi, Som

2017-10-01

The current study evaluated the interactive effects of chronic waterborne copper (Cu) and nickel (Ni) exposure on tissue-specific metal accumulation and reproductive performance in fathead minnow (Pimephales promelas). Fish trios (1 male: 2 female; n = 5-6) were exposed for 21 days to: (i) control (no added Cu or Ni), (ii) waterborne Cu (45 μg/L), (iii) waterborne Ni (270 μg/L), and (iv) binary mixture of waterborne Cu and Ni (45 and 270 μg/L, respectively). Fish fecundity (cumulative egg production) was found to be the most sensitive reproductive endpoint, and the interaction of Cu and Ni elicited an additive effect on egg production. Tissue-specific accumulation of both metals was not influenced by the interaction of Cu and Ni, except an increased Cu and Ni burden in the carcass and ovary, respectively, were recorded. The expressions of hepatic estrogen receptor genes (ER-α and ER-β) and the circulating estradiol level in females were also not affected by the metal-mixture treatment. However, co-exposure to waterborne Cu and Ni resulted in a significant downregulation of the hepatic vitellogenin gene in females, which was associated with the maximum upregulation of the hepatic metallothionein gene. In addition, a significant alteration of ovarian histopathology (decreased abundance of post-vitellogenic follicles, and increased follicular atresia) was also observed only in females exposed to Cu and Ni in mixture. Collectively, these observations suggest that chronic waterborne exposure to Cu and Ni in binary mixture may impair fish reproductive capacity by inducing histopathological damage in ovarian tissue, and disrupting of energy homeostasis in fish. Copyright © 2017 Elsevier Ltd. All rights reserved.

Xylella fastidiosa differentially accumulates mineral elements in biofilm and planktonic cells.

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Paul A Cobine

Full Text Available Xylella fastidiosa is a bacterial plant pathogen that infects numerous plant hosts. Disease develops when the bacterium colonizes the xylem vessels and forms a biofilm. Inductively coupled plasma optical emission spectroscopy was used to examine the mineral element content of this pathogen in biofilm and planktonic states. Significant accumulations of copper (30-fold, manganese (6-fold, zinc (5-fold, calcium (2-fold and potassium (2-fold in the biofilm compared to planktonic cells were observed. Other mineral elements such as sodium, magnesium and iron did not significantly differ between biofilm and planktonic cells. The distribution of mineral elements in the planktonic cells loosely mirrors the media composition; however the unique mineral element distribution in biofilm suggests specific mechanisms of accumulation from the media. A cell-to-surface attachment assay shows that addition of 50 to 100 µM Cu to standard X. fastidiosa media increases biofilm, while higher concentrations (>200 µM slow cell growth and prevent biofilm formation. Moreover cell-to-surface attachment was blocked by specific chelation of copper. Growth of X. fastidiosa in microfluidic chambers under flow conditions showed that addition of 50 µM Cu to the media accelerated attachment and aggregation, while 400 µM prevented this process. Supplementation of standard media with Mn showed increased biofilm formation and cell-to-cell attachment. In contrast, while the biofilm accumulated Zn, supplementation to the media with this element caused inhibited growth of planktonic cells and impaired biofilm formation. Collectively these data suggest roles for these minerals in attachment and biofilm formation and therefore the virulence of this pathogen.

Industrial Tests to Modify Molten Copper Slag for Improvement of Copper Recovery

Science.gov (United States)

Guo, Zhengqi; Zhu, Deqing; Pan, Jian; Zhang, Feng; Yang, Congcong

2018-04-01

In this article, to improve the recovery of copper from copper slag by flotation process, industrial tests of the modification process involving addition of a composite additive into molten copper slag were conducted, and the modified slag was subjected to the flotation process to confirm the modification effect. The phase evolution of the slag in the modification process was revealed by thermodynamic calculations, x-ray diffraction, optical microscopy and scanning electron microscopy. The results show that more copper was transformed and enriched in copper sulfide phases. The magnetite content in the modified slag decreased, and that of "FeO" increased correspondingly, leading to a better fluidity of the molten slag, which improved the aggregation and growth of fine particles of the copper sulfide minerals. Closed-circuit flotation tests of the original and modified slags were conducted, and the results show that the copper recovery increased obviously from 69.15% to 73.38%, and the copper grade of concentrates was elevated slightly from 20.24% to 21.69%, further confirming that the industrial tests of the modification process were successful. Hence, the modification process has a bright future in industrial applications for enhancing the recovery of copper from the copper slag.

Accumulation of heavy metals in freshwater fish in cage aquaculture at Cirata Reservoir, West Java, Indonesia.

Science.gov (United States)

Salami, Indah Rachmatiah Siti; Rahmawati, Suphia; Sutarto, Ratri Indri Hapsari; Jaya, Pinilih Marendra

2008-10-01

Freshwater fish demand in West Java is supported by cage aquaculture in reservoirs. Cirata Reservoir is one of three cascading hydropower reservoirs built along the Citarum River that receives domestic and industrial wastes. Water pollution by heavy metals, such as copper and lead, increases the health risk of humans who consume the fish. The study was aimed to evaluate the concentration of copper and lead in fish, specifically on Cyprinus carpio, that are widely cultivated in the Cirata Reservoir. Two sizes of fish were collected from five sampling points around floating-cage area. The liver of the fish was found to contain the highest copper level, followed by gills, skin, and muscle. In contrast, lead was also high in the liver, followed by skin, gills, and the muscle, but the copper concentration found in the tissues studied was much higher (31.111 +/- 17.911 mg/kg dry wt) than the lead content (0.290 +/- 0.346 mg/kg dry wt). The concentration of metals in smaller fish was always higher than than that in bigger fish. The metals content was also compared to those in fish exposed in a semistatic laboratory-scale study using Oreochromis niloticus. After 28 days exposure with 0.01 mg Cu/L, fish accumulated 21.53 mg Cu/kg dry wt, whereas for Pb exposure of 0.016 mg/L, fish accumulated up to 7 mg/kg dry wt. However, estimates of Cu and Pb intake from C. carpio consumption were still below the average daily intake (ADI) limit. This study suggested that monitoring of water quality and heavy metals in cultured fish is important in protecting human health.

Different toxicity mechanisms between bare and polymer-coated copper oxide nanoparticles in Lemna gibba

International Nuclear Information System (INIS)

Perreault, François; Popovic, Radovan; Dewez, David

2014-01-01

In this report, we investigated how the presence of a polymer shell (poly(styrene-co-butyl acrylate) alters the toxicity of CuO NPs in Lemna gibba. Based on total Cu concentration, core–shell CuO NPs were 10 times more toxic than CuO NPs, inducing a 50% decrease of growth rate at 0.4 g l −1 after 48-h of exposure while a concentration of 4.5 g l −1 was required for CuO NPs for a similar effect. Toxicity of CuO NPs was mainly due to NPs solubilization in the media. Based on the accumulated copper content in the plants, core–shell CuO NPs induced 4 times more reactive oxygen species compared to CuO NPs and copper sulfate, indicating that the presence of the polymer shell changed the toxic effect induced in L. gibba. This effect could not be attributed to the polymer alone and reveals that surface modification may change the nature of NPs toxicity. -- Highlights: • Bare and polymer-coated CuO nanoparticles were toxic to Lemna gibba. • Toxicity of bare CuO was mainly due to solubilized soluble copper. • Coated CuO accumulated inside the plants four times more. • Formation of reactive oxygen species was increased by polymer coating. • Coating of nanomaterials modifies mechanisms of action at cellular level. -- Polymer coating increases oxidative stress effect by core–shell CuO nanoparticles

Copper nitrate redispersion to arrive at highly active silica-supported copper catalysts

NARCIS (Netherlands)

Munnik, P.|info:eu-repo/dai/nl/328228524; Wolters, M.|info:eu-repo/dai/nl/304829560; Gabrielsson, A.; Pollington, S.D.; Headdock, G.; Bitter, J.H.|info:eu-repo/dai/nl/160581435; de Jongh, P.E.|info:eu-repo/dai/nl/186125372; de Jong, K.P.|info:eu-repo/dai/nl/06885580X

2011-01-01

In order to obtain copper catalysts with high dispersions at high copper loadings, the gas flow rate and gas composition was varied during calcination of silica gel impregnated with copper nitrate to a loading of 18 wt % of copper. Analysis by X-ray diffraction (XRD), N2O chemisorption, and

The enzymatic and antioxidative stress response of Lemna minor to copper and a chloroacetamide herbicide.

Science.gov (United States)

Obermeier, Michael; Schröder, Christian A; Helmreich, Brigitte; Schröder, Peter

2015-12-01

Lemna minor L., a widely used model plant for toxicity tests has raised interest for its application to phytoremediation due to its rapid growth and ubiquitous occurrence. In rural areas, the pollution of water bodies with heavy metals and agrochemicals poses a problem to surface water quality. Among problematic compounds, heavy metals (copper) and pesticides are frequently found in water bodies. To establish duckweed as a potential plant for phytoremediation, enzymatic and antioxidative stress responses of Lemna minor during exposure to copper and a chloroacetamide herbicide were investigated in laboratory studies. The present study aimed at evaluating growth and the antioxidative and glutathione-dependent enzyme activity of Lemna plants and its performance in a scenario for phytoremediation of copper and a chloroacetamide herbicide. Lemna minor was grown in Steinberg medium under controlled conditions. Plants were treated with CuSO4 (ion conc. 50 and 100 μg/L) and pethoxamide (1.25 and 2.5 μg/L). Measurements following published methods focused on plant growth, oxidative stress, and basic detoxification enzymes. Duckweed proved to survive treatment with the respective concentrations of both pollutants very well. Its growth was inhibited scarcely, and no visible symptoms occurred. On the cellular basis, accumulation of O2(-) and H2O2 were detected, as well as stress reactions of antioxidative enzymes. Duckweed detoxification potential for organic pollutants was high and increased significantly with incubation. Pethoxamide was found to be conjugated with glutathione. Copper was accumulated in the fronds at high levels, and transient oxidative defense reactions were triggered. This work confirms the significance of L. minor for the removal of copper from water and the conjugation of the selective herbicide pethoxamide. Both organic and inorganic xenobiotics induced different trends of enzymatic and antioxidative stress response. The strong increase of stress

Search of a plant hiperacumuladora of copper

International Nuclear Information System (INIS)

Hernandez Colorado, R.R.

2003-01-01

The objective of this investigation was to find inside the flora of Costa Rica, a plant copper hiperacumuladora. Once identified the species, you would proceed to cultivate it in vitro, stops later to make experiments of copper absorption in vitro. If the selected species was capable of hiperacumular the metal, the influence of the agents quelantes sour etilendiaminotetraacetico would be investigated (EDTA) and citric acid, in the absorption of the metal. To carry out the investigation, vegetable species were collected inside the banana property Probana located in Limon and inside a coffee plantation in Carrizal de Alajuela. The dried vegetable samples were analyzed the copper content and in the same way, with collected floor samples of the surrounding area to the plants. As satisfactory results were not obtained, the species Tagetes foetidissima and Coccosypselum hirsutum were selected. The first one was selected with base in their metabolism, which allowed to synthesize sulfurated compounds. The second were selected with base in the available information in the literature about the aluminum hiperacumulation on the part of species belonging to the family Rubiaceae, to which it belongs. The species T. foetidissima and C. hirsutum, were cultivated in vitro stops later to make experiments of copper absorption. The results obtained starting from the experiments of copper absorption for T. foetidissima were of 3626 - 7402 mg kg -1 it has more than enough it bases dry, for an environment of concentration of the metal in the means of cultivation of 20-30 mg L -1 for a lapse of fifteen days. For this species an environment of accumulation of 4251 was obtained - 6481 mg kg -1 it has more than enough it bases dry, to 20 mg L -1 of the metal, when adding him EDTA to an experimentation group and citric acid to another. In the case of C. hirsutum, hiperacumulo gets paid in concentrations of 7648 - 8786 mg kg -1 it has more than enough it bases dry, for an environment of

Ecological significance of compatible solute accumulation by micro-organisms: from single cells to global climate.

Science.gov (United States)

Welsh, D T

2000-07-01

The osmoadaptation of most micro-organisms involves the accumulation of K(+) ions and one or more of a restricted range of low molecular mass organic solutes, collectively termed 'compatible solutes'. These solutes are accumulated to high intracellular concentrations, in order to balance the osmotic pressure of the growth medium and maintain cell turgor pressure, which provides the driving force for cell extension growth. In this review, I discuss the alternative roles which compatible solutes may also play as intracellular reserves of carbon, energy and nitrogen, and as more general stress metabolites involved in protection of cells against other environmental stresses including heat, desiccation and freezing. Thus, the evolutionary selection for the accumulation of a specific compatible solute may not depend solely upon its function during osmoadaptation, but also upon the secondary benefits its accumulation provides, such as increased tolerance of other environmental stresses prevalent in the organism's niche or even anti-herbivory or dispersal functions in the case of dimethylsulfoniopropionate (DMSP). In the second part of the review, I discuss the ecological consequences of the release of compatible solutes to the environment, where they can provide sources of compatible solutes, carbon, nitrogen and energy for other members of the micro-flora. Finally, at the global scale the metabolism of specific compatible solutes (betaines and DMSP) in brackish water, marine and hypersaline environments may influence global climate, due to the production of the trace gases, methane and dimethylsulfide (DMS) and in the case of DMS, also couple the marine and terrestrial sulfur cycles.

Intracellular HIV-1 Gag localization is impaired by mutations in the nucleocapsid zinc fingers

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Muriaux Delphine

2007-08-01

Full Text Available Abstract Background The HIV-1 nucleocapsid protein (NC is formed of two CCHC zinc fingers flanked by highly basic regions. HIV-1 NC plays key roles in virus structure and replication via its nucleic acid binding and chaperoning properties. In fact, NC controls proviral DNA synthesis by reverse transcriptase (RT, gRNA dimerization and packaging, and virion assembly. Results We previously reported a role for the first NC zinc finger in virion structure and replication 1. To investigate the role of both NC zinc fingers in intracellular Gag trafficking, and in virion assembly, we generated series of NC zinc fingers mutations. Results show that all Zinc finger mutations have a negative impact on virion biogenesis and maturation and rendered defective the mutant viruses. The NC zinc finger mutations caused an intracellular accumulation of Gag, which was found either diffuse in the cytoplasm or at the plasma membrane but not associated with endosomal membranes as for wild type Gag. Evidences are also provided showing that the intracellular interactions between NC-mutated Gag and the gRNA were impaired. Conclusion These results show that Gag oligomerization mediated by gRNA-NC interactions is required for correct Gag trafficking, and assembly in HIV-1 producing cells and the release of infectious viruses.

Earth's copper resources estimated from tectonic diffusion of porphyry copper deposits

Science.gov (United States)

Kesler, Stephen E.; Wilkinson, Bruce H.

2008-03-01

Improved estimates of global mineral endowments are relevantto issues ranging from strategic planning to global geochemicalcycling. We have used a time-space model for the tectonic migrationof porphyry copper deposits vertically through the crust tocalculate Earth's endowment of copper in mineral deposits. Themodel relies only on knowledge of numbers and ages of porphyrycopper deposits, Earth's most widespread and important sourceof copper, in order to estimate numbers of eroded and preserveddeposits in the crust. Model results indicate that 125,895 porphyrycopper deposits were formed during Phanerozoic time, that only47,789 of these remain at various crustal depths, and that thesecontain 1.7 x 1011 tonnes (t) of copper. Assuming that othertypes of copper deposits behave similarly in the crust and haveabundances proportional to their current global production yieldsan estimate of 3 x 1011 t for total global copper resourcesat all levels in Earth's crust. Thus, 0.25% of the copper inthe crust has been concentrated into deposits through Phanerozoictime, and about two-thirds of this has been recycled by upliftand erosion. The amount of copper in deposits above 3.3 km,a likely limit of future mining, could supply current worldmine production for 5500 yr, thus quantifying the highly unusualand nonrenewable nature of mineral deposits.

Intracellular accumulation of mannopine, an opine produced by crown gall tumors, transiently inhibits growth of Agrobacterium tumefaciens.

Science.gov (United States)

Kim, K S; Baek, C H; Lee, J K; Yang, J M; Farrand, S K

2001-06-01

pYDH208, a cosmid clone from the octopine-mannityl opine-type tumor-inducing (Ti) plasmid pTi15955 confers utilization of mannopine (MOP) and agropine (AGR) on Agrobacterium tumefaciens strain NT1. NT1 harboring pYDH208 with an insertion mutation in mocC, which codes for MOP oxidoreductase, not only fails to utilize MOP as a sole carbon source, but also was inhibited in its growth by MOP and AGR. In contrast, the growth of mutants with insertions in other tested moc genes was not inhibited by either opine. Growth of strains NT1 or UIA5, a derivative of C58 that lacks pAtC58, was not inhibited by MOP, but growth of NT1 or UIA5 harboring pRE10, which codes for the MOP transport system, was inhibited by the opine. When a clone expressing mocC was introduced, the growth of strain NT1(pRE10) was not inhibited by MOP, although UIA5(pRE10) was still weakly inhibited. In strain NT1(pRE10, mocC), santhopine (SOP), produced by the oxidation of MOP by MocC, was further degraded by functions encoded by pAtC58. These results suggest that MOP and, to a lesser extent, SOP are inhibitory when accumulated intracellularly. The growth of NT1(pRE10), as measured by turbidity and viable cell counts, ceased upon the addition of MOP but restarted in a few hours. Regrowth was partly the result of the outgrowth of spontaneous MOP-resistant mutants and partly the adaptation of cells to MOP in the medium. Chrysopine, isochrysopine, and analogs of MOP in which the glutamine residue is substituted with other amino acids were barely taken up by NT1(pRE10) and were not inhibitory to growth of the strain. Sugar analogs of MOP were inhibitory, and those containing sugars in the D form were more inhibitory than those containing sugars in the L form. MOP analogs containing hexose sugars were more inhibitory than those containing sugars with three, four, or five carbon atoms. Mutants of NT1(pRE10) that are resistant to MOP arose in the zone of growth inhibition. Genetic and physiological analyses

Insecticide resistance and intracellular proteases.

Science.gov (United States)

Wilkins, Richard M

2017-12-01

Pesticide resistance is an example of evolution in action with mechanisms of resistance arising from mutations or increased expression of intrinsic genes. Intracellular proteases have a key role in maintaining healthy cells and in responding to stressors such as pesticides. Insecticide-resistant insects have constitutively elevated intracellular protease activity compared to corresponding susceptible strains. This increase was shown for some cases originally through biochemical enzyme studies and subsequently putatively by transcriptomics and proteomics methods. Upregulation and expression of proteases have been characterised in resistant strains of some insect species, including mosquitoes. This increase in proteolysis results in more degradation products (amino acids) of intracellular proteins. These may be utilised in the resistant strain to better protect the cell from stress. There are changes in insect intracellular proteases shortly after insecticide exposure, suggesting a role in stress response. The use of protease and proteasome inhibitors or peptide mimetics as synergists with improved application techniques and through protease gene knockdown using RNA interference (possibly expressed in crop plants) may be potential pest management strategies, in situations where elevated intracellular proteases are relevant. © 2017 Society of Chemical Industry. © 2017 Society of Chemical Industry.

Effects of dust accumulation and removal on radiator surfaces on Mars

International Nuclear Information System (INIS)

Gaier, J.R.; Perez-Davis, M.E.; Rutledge, S.K.; Hotes, D.; Olle, R.

1991-01-01

Tests were carried out to assess the impact of wind blown dust accumulation and abrasion on radiator surfaces on Mars. High emittance arc-textured copper (Cu) and niobium-1%-zirconium (Nb-1%Zr) samples were subjected to basaltic dust laden wind at Martian pressure (1000 Pa) at speeds varying from 19 to 97 m/s in the Martian Surface Wind Tunnel at NASA Ames Research Center. The effect of accumulated dust was also observed by pre-dusting some of the samples before the test. Radiator degradation was determined by measuring the change in the emittance after dust was deposited and/or removed. The principal mode of degradation was abrasion. Arc-textured Nb-1%Zr proved to be more susceptible to degradation than Cu, and pre-dusting appeared to have lessened the abrasion

Genome Sequences of Two Copper-Resistant Escherichia coli Strains Isolated from Copper-Fed Pigs

DEFF Research Database (Denmark)

Lüthje, Freja L.; Hasman, Henrik; Aarestrup, Frank Møller

2014-01-01

The draft genome sequences of two copper-resistant Escherichia coli strains were determined. These had been isolated from copper-fed pigs and contained additional putative operons conferring copper and other metal and metalloid resistances.......The draft genome sequences of two copper-resistant Escherichia coli strains were determined. These had been isolated from copper-fed pigs and contained additional putative operons conferring copper and other metal and metalloid resistances....

Copper uptake and retention in liver parenchymal cells isolated from nutritionally copper-deficient rats

NARCIS (Netherlands)

Berg, van den G.J.; de Goeij, J.J.M.; Bock, I.; Gijbels, M.J.J.; Brouwer, A.; Lei, K.Y.; Hendriks, H.F.J.

1991-01-01

Copper uptake and retention were studied in primary cultures of liver parenchymal cells isolated from copper-deficient rats. Male Sprague-Dawley rats were fed a copper-deficient diet (<1 mg Cu/kg) for 10 wk. Copper-deficient rats were characterized by low copper concentrations in plasma and liver,

Copper uptake and retention in liver parenchymal cells isolated from nutritionally copper-deficient rats

NARCIS (Netherlands)

Berg, G.J. van den; Goeij, J.J.M. de; Bock, I.; Gijbels, M.J.J.; Brouwer, A.; Lei, K.Y.; Hendruiks, H.F.J.

1991-01-01

Copper uptake and retention were studied in primary cultures of liver parenchymal cells isolated from copper-deficient rats. Male Sprague-Dawley rats were fed a copper-deficient diet (< 1 mg Cu/kg) for 10 wk. Copper-deficient rats were characterized by low copper concentrations in plasma and liver,

Studies performed on neutron-irradiated copper-doped iron specimens by means of neutron small-angle scattering

International Nuclear Information System (INIS)

Naraghi, M.

1978-01-01

By means of neutron small-angle scattering precipitation arising from heat-treatment and reactor irradiation in copper-alloyed iron specimens were studied. Copper content varried between 0 and 1.5%, irradiation temperature between 310 and 563K. The specimens had been cooled from the melt partly fast, partly slowely. By taking account of magnetic scattering and by investigating the azimuthal dependence of the total scattering it became possible to distinguish between copper precipitations and vacancy agglomerates. The most obvious effect in the slowly cooled specimens after irradiation with 2-10 19 fast neutrons per cm 2 at a temperature of 563 K is the existence of copper agglomerates with diameters of the order of magnitude of 5nm. Precipitation already occurs to a much lesser extent by the influence of temperature alone. Fast cooling from the melt or low irradiation temperature reduce precipitation during reactor irradiation. Moreover, there are indications on the formation of vacancy accumulations and dislocation rings, the latter especially in the fast cooled specimens. (orig.) [de

New data on the Hyrkkoelae U-Cu mineralization: the behaviour of native copper in a natural environment

Energy Technology Data Exchange (ETDEWEB)

Marcos, N. [Helsinki Univ. of Technology, Espoo (Finland); Ahonen, L. [Geological Survey of Finland, Helsinki (Finland)

1999-05-01

The Hyrkkoelae Cu-U mineralisation (SW Finland) is studied as an analogue to the behaviour of copper canister in crystalline bedrock. Uranium-native copper and uranium-copper corrosion products interactions are also addressed in this study. The integration of uranium series disequilibrium (USD) studies gives an estimate of the time-scales of the corrosion processes. The mineral assemblages native copper-copper sulfide, copper sulfides - copper iron sulfides, and native copper-copper oxide (cuprite) occur in open fractures at several depth intervals within granite pegmatites (GP). The surfaces of these open fractures have accumulations of uranophane crystals and other unidentified uranyl compounds. The secondary uranium minerals are mainly distributed around copper sulfide grains. Microscopic intergrowths of copper sulfides and uranyl compounds also have been observed. The surface of the fracture where native copper and cuprite occur is covered with uranium-rich smectite. The very low {sup 234}U/{sup 238}U activity ratio (0.29 - 0.39) in the main uranium fraction in smectite indicates chemical stable conditions (e.g., oxidising) during at least a time period comparable to the half-life of the {sup 234}U isotope (T{sub 1/2} = 2.44 x 10{sup 5} a). Groundwater samples were collected from intervals where copper minerals occur within open fractures. The Eh and pH conditions were measured during long-term pumping (2-4 weeks per sample). Eh was measured both in situ and an the surface using three electrodes (Pt, Au, C). The actual groundwater conditions are oxidising and would not allow the sulfidization of native copper. Sulfidization may be considered as on old phenomenon, older than the precipitation of uranyl phases in the samples. The end of sulfidization may be earlier than the precipitation and/or remobilisation of U(VI) phases in a time span from about 2 x 10{sup 5} years (precipitation of uranophane) to 2.44 x 10{sup 5} (remobilisation of U from smectite). (orig.)

New data on the Hyrkkoelae U-Cu mineralization: the behaviour of native copper in a natural environment

International Nuclear Information System (INIS)

Marcos, N.; Ahonen, L.

1999-05-01

The Hyrkkoelae Cu-U mineralisation (SW Finland) is studied as an analogue to the behaviour of copper canister in crystalline bedrock. Uranium-native copper and uranium-copper corrosion products interactions are also addressed in this study. The integration of uranium series disequilibrium (USD) studies gives an estimate of the time-scales of the corrosion processes. The mineral assemblages native copper-copper sulfide, copper sulfides - copper iron sulfides, and native copper-copper oxide (cuprite) occur in open fractures at several depth intervals within granite pegmatites (GP). The surfaces of these open fractures have accumulations of uranophane crystals and other unidentified uranyl compounds. The secondary uranium minerals are mainly distributed around copper sulfide grains. Microscopic intergrowths of copper sulfides and uranyl compounds also have been observed. The surface of the fracture where native copper and cuprite occur is covered with uranium-rich smectite. The very low 234 U/ 238 U activity ratio (0.29 - 0.39) in the main uranium fraction in smectite indicates chemical stable conditions (e.g., oxidising) during at least a time period comparable to the half-life of the 234 U isotope (T 1/2 = 2.44 x 10 5 a). Groundwater samples were collected from intervals where copper minerals occur within open fractures. The Eh and pH conditions were measured during long-term pumping (2-4 weeks per sample). Eh was measured both in situ and an the surface using three electrodes (Pt, Au, C). The actual groundwater conditions are oxidising and would not allow the sulfidization of native copper. Sulfidization may be considered as on old phenomenon, older than the precipitation of uranyl phases in the samples. The end of sulfidization may be earlier than the precipitation and/or remobilisation of U(VI) phases in a time span from about 2 x 10 5 years (precipitation of uranophane) to 2.44 x 10 5 (remobilisation of U from smectite). (orig.)

Annual Copper Mountain Conferences on Multigrid and Iterative Methods, Copper Mountain, Colorado

International Nuclear Information System (INIS)

McCormick, Stephen F.

2016-01-01

This project supported the Copper Mountain Conference on Multigrid and Iterative Methods, held from 2007 to 2015, at Copper Mountain, Colorado. The subject of the Copper Mountain Conference Series alternated between Multigrid Methods in odd-numbered years and Iterative Methods in even-numbered years. Begun in 1983, the Series represents an important forum for the exchange of ideas in these two closely related fields. This report describes the Copper Mountain Conference on Multigrid and Iterative Methods, 2007-2015. Information on the conference series is available at http://grandmaster.colorado.edu/~copper/

Annual Copper Mountain Conferences on Multigrid and Iterative Methods, Copper Mountain, Colorado

Energy Technology Data Exchange (ETDEWEB)

McCormick, Stephen F. [Front Range Scientific, Inc., Lake City, CO (United States)

2016-03-25

This project supported the Copper Mountain Conference on Multigrid and Iterative Methods, held from 2007 to 2015, at Copper Mountain, Colorado. The subject of the Copper Mountain Conference Series alternated between Multigrid Methods in odd-numbered years and Iterative Methods in even-numbered years. Begun in 1983, the Series represents an important forum for the exchange of ideas in these two closely related fields. This report describes the Copper Mountain Conference on Multigrid and Iterative Methods, 2007-2015. Information on the conference series is available at http://grandmaster.colorado.edu/~copper/.

Optimal copper supply is required for normal plant iron deficiency responses

OpenAIRE

Waters, Brian M; Armbrust, Laura C

2013-01-01

Iron (Fe) and copper (Cu) homeostasis are tightly linked across biology. Understanding crosstalk between Fe and Cu nutrition could lead to strategies for improved growth on soils with low or excess metals, with implications for agriculture and phytoremediation. Here, we show that Cu and Fe nutrition interact to increase or decrease Fe and/or Cu accumulation in leaves and Fe uptake processes. Leaf Cu concentration increased under low Fe supply, while high Cu lowered leaf Fe concentration. Ferr...

Copper wire bonding

CERN Document Server

Chauhan, Preeti S; Zhong, ZhaoWei; Pecht, Michael G

2014-01-01

This critical volume provides an in-depth presentation of copper wire bonding technologies, processes and equipment, along with the economic benefits and risks.  Due to the increasing cost of materials used to make electronic components, the electronics industry has been rapidly moving from high cost gold to significantly lower cost copper as a wire bonding material.  However, copper wire bonding has several process and reliability concerns due to its material properties.  Copper Wire Bonding book lays out the challenges involved in replacing gold with copper as a wire bond material, and includes the bonding process changes—bond force, electric flame off, current and ultrasonic energy optimization, and bonding tools and equipment changes for first and second bond formation.  In addition, the bond–pad metallurgies and the use of bare and palladium-coated copper wires on aluminum are presented, and gold, nickel and palladium surface finishes are discussed.  The book also discusses best practices and re...

Mechanisms of metal tolerance in marine macroalgae, with emphasis on copper tolerance in Chlorophyta and Rhodophyta.

Science.gov (United States)

Moenne, Alejandra; González, Alberto; Sáez, Claudio A

2016-07-01

Green and red macroalgae are closely related organisms, and with terrestrial plants, and constitute the base of marine food webs in coastal ecosystems. Green and red seaweeds, as all living organisms, require essential metals, such as copper, iron, zinc, which can act as co-factors for several proteins and enzymes; however, these metals in excess can induce stress and impair cell viability. Most important negative effects of metal excess are related to the induction of an oxidative stress condition, characterized by the over-accumulation of Reactive Oxygen Species (ROS). In this respect, copper, abundant in wastewaters disposed to coastal environments from domestic and industrial activities, has been one of the most studied metals. Different investigations have provided evidence that green and red macroalgae display several defenses against copper excess to prevent, or at least reduce, stress and damage, among which are cellular exclusion mechanisms, synthesis of metal-chelating compounds, and the activation of the antioxidant system. Most important defense mechanisms identified in green and red seaweed involve: metal-binding to cell wall and epibionts; syntheses of metallothioneins and phytochelatins that accumulate in the cytoplasm; and the increase in the activity of antioxidant enzymes such as superoxide dismutase, ascorbate peroxidase, glutathione peroxidase and catalase, and greater production of antioxidant metabolites as glutathione and ascorbate in organelles and the cytoplasm. In this review, we go through historical records, latest advances, and pending tasks aiming to expand our current knowledge on defense mechanisms to copper excess in green and red macroalgae, with emphasis on biochemical and molecular aspects. Copyright © 2016 Elsevier B.V. All rights reserved.

Heavy metal and nutrient uptake in plants colonizing post-flotation copper tailings.

Science.gov (United States)

Kasowska, Dorota; Gediga, Krzysztof; Spiak, Zofia

2018-01-01

Copper ore mining and processing release hazardous post-flotation wastes that are difficult for remediation. The studied tailings were extremely rich in Cu (1800 mg kg -1 ) and contaminated with Co and Mn, and contained very little available forms of P, Fe, and Zn. The plants growing in tailings were distinctly enriched in Cu, Cd, Co, Ni, and Pb, and the concentration of copper achived the critical toxicity level in shoots of Cerastium arvense and Polygonum aviculare. The redundancy analysis demonstrated significant relationship between the concentration of available forms of studied elements in substrate and the chemical composition of plant shoots. Results of the principal component analysis enabled to distinguish groups of plants which significantly differed in the pattern of element accumulation. The grass species Agrostis stolonifera and Calamagrostis epigejos growing in the tailings accumulated significantly lower amounts of Cu, but they also had the lowest levels of P, Fe, and Zn in comparison to dicotyledonous. A. stolonifera occurred to be the most suitable species for phytostabilization of the tailings with regard to its low shoot Cu content and more efficient acquisition of limiting nutrients in relation to C. epigejos. The amendments improving texture, phosphorus fertilization, and the introduction of native leguminous species were recommended for application in the phytoremediation process of the tailings.

Glassy carbon electrode modified with polyanilne/ethylenediamine for detection of copper ions

Science.gov (United States)

Patil, Harshada K.; Deshmukh, Megha A.; Bodkhe, Gajanan A.; Shirsat, Mahendra D.

2018-05-01

Increasing water pollution is having high concern, since it creates the threats to all leaving organisms of existence. Industrial sewages have not only polluted the main stream lines of water, also the ground level water is having serious contaminations. Heavy metal ions are the pollutants which are not degradable and can be accumulated on living things ultimately the excess accumulation results into the serious concerns. Therefore, it is necessary to develop the sensors which can detect the heavy metal ions up to its maximum contamination limits. Conducting polymers are the materials which possess large application spectra. This investigation reports the electrochemically synthesized polyaniline (PANI) for modification of glassy carbon electrode (GCE). Ethylenediamine (EDA) - chelating ligand used for the modification of polyaniline so as to inculcate the selectivity toward copper ions Cu (II). The electrochemical cyclic voltammetry (CV) was used for the study of redox characteristics of PANI and influence of EDA modification. The result of CV has shown the reduced oxidation and reduction peak currents after modification indicating the domination of EDA. GCE modified with PANI/EDA was then employed for the detection of divalent copper ions and have shown the affinity toward Cu ions. The detection limit achieved was equal to 10mg/lit.

Tritium Suicide Selection Identifies Proteins Involved in the Uptake and Intracellular Transport of Sterols in Saccharomyces cerevisiae▿

Science.gov (United States)

Sullivan, David P.; Georgiev, Alexander; Menon, Anant K.

2009-01-01

Sterol transport between the plasma membrane (PM) and the endoplasmic reticulum (ER) occurs by a nonvesicular mechanism that is poorly understood. To identify proteins required for this process, we isolated Saccharomyces cerevisiae mutants with defects in sterol transport. We used Upc2-1 cells that have the ability to take up sterols under aerobic conditions and exploited the observation that intracellular accumulation of exogenously supplied [3H]cholesterol in the form of [3H]cholesteryl ester requires an intact PM-ER sterol transport pathway. Upc2-1 cells were mutagenized using a transposon library, incubated with [3H]cholesterol, and subjected to tritium suicide selection to isolate mutants with a decreased ability to accumulate [3H]cholesterol. Many of the mutants had defects in the expression and trafficking of Aus1 and Pdr11, PM-localized ABC transporters that are required for sterol uptake. Through characterization of one of the mutants, a new role was uncovered for the transcription factor Mot3 in controlling expression of Aus1 and Pdr11. A number of mutants had transposon insertions in the uncharacterized Ydr051c gene, which we now refer to as DET1 (decreased ergosterol transport). These mutants expressed Aus1 and Pdr11 normally but were severely defective in the ability to accumulate exogenously supplied cholesterol. The transport of newly synthesized sterols from the ER to the PM was also defective in det1Δ cells. These data indicate that the cytoplasmic protein encoded by DET1 is involved in intracellular sterol transport. PMID:19060182

Cardiac lipid accumulation associated with diastolic dysfunction in obese mice

DEFF Research Database (Denmark)

Christoffersen, Christina; Bollano, Entela; Lindegaard, Marie L S

2003-01-01

Obesity may confer cardiac dysfunction due to lipid accumulation in cardiomyocytes. To test this idea, we examined whether obese ob/ob mice display heart lipid accumulation and cardiac dysfunction. Ob/ob mouse hearts had increased expression of genes mediating extracellular generation, transport....../ob mice and 2.5 +/- 0.1 in ob/+ mice (P = 0.0001). In contrast, the indexes of systolic function and heart brain natriuretic peptide mRNA expression were only marginally affected and unaffected, respectively, in ob/ob compared with ob/+ mice. The results suggest that ob/ob mouse hearts have increased...... across the myocyte cell membrane, intracellular transport, mitochondrial uptake, and beta-oxidation of fatty acids compared with ob/+ mice. Accordingly, ob/ob mouse hearts contained more triglyceride (6.8 +/- 0.4 vs. 2.3 +/- 0.4 microg/mg; P hearts. Histological examinations...

Removal of Copper by Eichhornia crassipes and the Characterization of Associated Bacteria of the Rhizosphere System

Directory of Open Access Journals (Sweden)

Raisa Kabeer

2014-06-01

Full Text Available Excess doses of trace element contamination make conventional water treatment methods less effective and more expensive, where in alternative biotechnological applications open up new opportunities with their reduced cost and lesser impacts to the environment. In the present investigation, effectiveness of aquatic macrophyte Eichhornia crassipes was tested for the removal of copper in laboratory conditions. Water samples were collected from macrophytes natural habitat and water tubs used for growing E. crassipes and analysed along with plant tissues for Cu content. The work also characterized the associated microbiota of the rhizosphere system of the E. crassipes as well as the wetland system of its occurrence. Copper concentration of the wetland water samples ranged from 0.009 to 0.03ppm. Six bacterial genera (Acinetobacter, Alcaligenes, Bacillus, Kurthia, Listeria and Chromobacterium were represented in rhizosphere of E.crassipes and 4 bacterial genera (Acinetobacter, Bacillus, Listeria and Chromobacterium were recorded in wetland water samples. Copper resistance studies of the bacterial isolates showed that out of 26 isolates from rhizosphere and 19 strains from water samples,12 of them showed low resistance (80% of copper during 15 days experiment. Copper accumulation was found to be high in the root followed by leaf and petiole. Results of the present study concluded that E. crassipes is an efficient plant for the removal of copper.

Nearly 60% Copper Rod & Wire Companies Neutral about Future Copper Price

Institute of Scientific and Technical Information of China (English)

2012-01-01

<正>How about the trend of copper price recently? According to the survey result of Shanghai Metals Market, amongst 21 domestic copper rod & wire companies, 57% of the companies are neutral about the future copper price, while 14% and 19% of the companies consider that

Disposal and improvement of contaminated by waste extraction of copper mining in chile

Science.gov (United States)

Naranjo Lamilla, Pedro; Blanco Fernández, David; Díaz González, Marcos; Robles Castillo, Marcelo; Decinti Weiss, Alejandra; Tapia Alvarez, Carolina; Pardo Fabregat, Francisco; Vidal, Manuel Miguel Jordan; Bech, Jaume; Roca, Nuria

2016-04-01

This project originated from the need of a mining company, which mines and processes copper ore. High purity copper is produced with an annual production of 1,113,928 tons of concentrate to a law of 32%. This mining company has generated several illegal landfills and has been forced by the government to make a management center Industrial Solid Waste (ISW). The forecast volume of waste generated is 20,000 tons / year. Chemical analysis established that the studied soil has a high copper content, caused by nature or from the spread of contaminants from mining activities. Moreover, in some sectors, soil contamination by mercury, hydrocarbons and oils and fats were detected, likely associated with the accumulation of waste. The waters are also impacted by mining industrial tasks, specifically copper ores, molybdenum, manganese, sulfates and have an acidic pH. The ISW management center dispels the pollution of soil and water and concentrating all activities in a technically suitable place. In this center the necessary guidelines for the treatment and disposal of soil contamination caused by uncontrolled landfills are given, also generating a leachate collection system and a network of fluid monitoring physicochemical water quality and soil environment. Keywords: Industrial solid waste, soil contamination, Mining waste

Roadside soils show low plant available zinc and copper concentrations

International Nuclear Information System (INIS)

Morse, Natalie; Walter, M. Todd; Osmond, Deanna; Hunt, William

2016-01-01

Vehicle combustion and component wear are a major source of metal contamination in the environment, which could be especially concerning where road ditches are actively farmed. The objective of this study was to assess how site variables, namely age, traffic (vehicles day"−"1), and percent carbon (%C) affect metal accumulation in roadside soils. A soil chronosequence was established with sites ranging from 3 to 37 years old and bioavailable, or mobile, concentrations of Zinc (Zn) and Copper (Cu) were measured along major highways in North Carolina using a Mehlich III extraction. Mobile Zn and Cu concentrations were low overall, and when results were scaled via literature values to “total metal”, the results were still generally lower than previous roadside studies. This could indicate farming on lands near roads would pose a low plant toxicity risk. Zinc and Cu were not correlated with annual average traffic count, but were positively correlated with lifetime traffic load (the product of site age and traffic count). This study shows an often overlooked variable, site age, should be included when considering roadside pollution accumulation. Zinc and Cu were more strongly associated with %C, than traffic load. Because vehicle combustion is also a carbon source, it is not obvious whether the metals and carbon are simply co-accumulating or whether the soil carbon in roadside soils may facilitate previously overlooked roles in sequestering metals on-site. - Highlights: • Low plant available zinc and copper concentrations in roadside soils of the southeast U.S. • Metals from vehicular traffic may not be adversely affecting plants in roadside environment. • Traffic volume and site age better predictor of metal pollution than traffic volume alone. - Mobile concentrations of Zn and Cu in roadside soils were below toxic levels. Zn and Cu concentrations were better correlated with lifetime vehicle load, as opposed to traffic volume.

Study on accumulation mechanism of Cu and Pb in moss, Scopelophila cataractae, by micro-XRF imaging and XAFS analysis

International Nuclear Information System (INIS)

Yoshii, Yuichi; Nakai, Izumi; Hokura, Akiko; Abe, Tomoko; Itouga, Misao; Sakakibara, Hitoshi; Terada, Yasuko

2011-01-01

Scopelophila cataractae is known as a 'copper moss'. It is known as a peculiar moss which can accumulate high level of metals (Cu, Pb, etc.) in its plant body. This moss is expected as a material to collect metals from water. In this study, gametophyte's leaf of the moss was examined by μ-XRF imaging, and Cu K-edge and Pb L 1 -, L 3 -edge XAFS analyses to reveal the accumulation behavior of Cu and Pb. μ-XRF imaging was carried out utilizing X-ray microbeam at BL37XU, SPring-8 from the undulator source, and at BL-4A, Photon Factory KEK from the bending magnet. The results have revealed that Cu and Pb were accumulated on the cell wall of the leaf, and especially at the stereid cells of midrib. It was found that Cu-XANES spectra of the leaf were similar to that of copper(II) acetate, a reference substance. On the other hand, Pb XANES spectra of the leaf were similar to that of a reference, lead(II) stearate. These results suggested that these metals were bound to oxygen of carboxyl group in the plant. Consequently, it is assumed that Cu and Pb could be bound to acidic sugar which is a component of its cell wall such as pectic acid, polygalacturonic acid and others. Furthermore, to compare with other heavy-metal accumulating moss (Scopelophila ligulata and Sphagnum palustre), their gametophyte's leaves were also examined by μ-XRF imaging. S. ligulata accumulated Pb in its stereid cells of midlib, while S. palustre accumulated Pb in its hyaline cells. (author)

Zinc deficiency-induced iron accumulation, a consequence of alterations in iron regulatory protein-binding activity, iron transporters, and iron storage proteins.

Science.gov (United States)

Niles, Brad J; Clegg, Michael S; Hanna, Lynn A; Chou, Susan S; Momma, Tony Y; Hong, Heeok; Keen, Carl L

2008-02-22

One consequence of zinc deficiency is an elevation in cell and tissue iron concentrations. To examine the mechanism(s) underlying this phenomenon, Swiss 3T3 cells were cultured in zinc-deficient (D, 0.5 microM zinc), zinc-supplemented (S, 50 microM zinc), or control (C, 4 microM zinc) media. After 24 h of culture, cells in the D group were characterized by a 50% decrease in intracellular zinc and a 35% increase in intracellular iron relative to cells in the S and C groups. The increase in cellular iron was associated with increased transferrin receptor 1 protein and mRNA levels and increased ferritin light chain expression. The divalent metal transporter 1(+)iron-responsive element isoform mRNA was decreased during zinc deficiency-induced iron accumulation. Examination of zinc-deficient cells revealed increased binding of iron regulatory protein 2 (IRP2) and decreased binding of IRP1 to a consensus iron-responsive element. The increased IRP2-binding activity in zinc-deficient cells coincided with an increased level of IRP2 protein. The accumulation of IRP2 protein was independent of zinc deficiency-induced intracellular nitric oxide production but was attenuated by the addition of the antioxidant N-acetylcysteine or ascorbate to the D medium. These data support the concept that zinc deficiency can result in alterations in iron transporter, storage, and regulatory proteins, which facilitate iron accumulation.

The effects of silver ions on copper metabolism in rats.

Science.gov (United States)

Ilyechova, E Yu; Saveliev, A N; Skvortsov, A N; Babich, P S; Zatulovskaia, Yu A; Pliss, M G; Korzhevskii, D E; Tsymbalenko, N V; Puchkova, L V

2014-10-01

The influence of short and prolonged diet containing silver ions (Ag-diet) on copper metabolism was studied. Two groups of animals were used: one group of adult rats received a Ag-diet for one month (Ag-A1) and another group received a Ag-diet for 6 months from birth (Ag-N6). In Ag-A1 rats, the Ag-diet caused a dramatic decrease of copper status indexes that was manifested as ceruloplasmin-associated copper deficiency. In Ag-N6 rats, copper status indexes decreased only 2-fold as compared to control rats. In rats of both groups, silver entered the bloodstream and accumulated in the liver. Silver was incorporated into ceruloplasmin (Cp), but not SOD1. In the liver, a prolonged Ag-diet caused a decrease of the expression level of genes, associated with copper metabolism. Comparative spectrophotometric analysis of partially purified Cp fractions has shown that Cp from Ag-N6 rats was closer to holo-Cp by specific enzymatic activities and tertiary structure than Cp from Ag-A1 rats. However, Cp of Ag-N6 differs from control holo-Cp and Cp of Ag-A1 in its affinity to DEAE-Sepharose and in its binding properties to lectins. In the bloodstream of Ag-N6, two Cp forms are present as shown in pulse-experiments on rats with the liver isolated from circulation. One of the Cp isoforms is of hepatic origin, and the other is of extrahepatic origin; the latter is characterized by a faster rate of secretion than hepatic Cp. These data allowed us to suggest that the disturbance of holo-Cp formation in the liver was compensated by induction of extrahepatic Cp synthesis. The possible biological importance of these effects is discussed.

HALOGENATED AROMATIC HYDROCARBON-MEDIATED PORPHYRIN ACCUMULATION AND INDUCTION OF CYTOCHROME P4501A IN CHICKEN EMBRYO HEPATOCYTES. (R823889)

Science.gov (United States)

Concentration-dependent induction of cytochrome P4501A (CYP1A) and intracellular porphyrin accumulation were observed following treatment of chicken embryo hepatocyte (CEH) cultures with 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD), 2,3,7,8-tetrachlorodibenzofuran (TCDF), 3,3',4,4'...

Accumulation and distribution of heavy metals in the grey mangrove, Avicennia marina (Forsk.)Vierh.: biological indication potential

International Nuclear Information System (INIS)

MacFarlane, G.R.; Pulkownik, A.; Burchett, M.D.

2003-01-01

Mangrove roots may serve as a sensitive bio-indicator for metal pollution in estuarine systems. - The accumulative partitioning of the heavy metals Cu, Pb and Zn in the grey mangrove, Avicennia marina, were studied under field conditions. Copper and Pb were accumulated in root tissue to levels higher than surrounding sediment levels. Zinc was accumulated to levels reflecting sediment concentrations. Strong linear relationships existed for all metals in sediments with metals in root tissue. Accumulation of Cu in leaf tissue followed a linear relationship at lower sediment concentrations, with an exclusion or saturation mechanism at higher sediment concentrations. Lead showed little mobility to leaf tissue. Zn showed restricted accumulation in leaf tissue, which correlated with sediment concentrations. Decreases in sediment pH were found to increase Zn accumulation to root tissue. Increasing concentrations of Pb and Zn in sediments resulted in a greater accumulation of Pb to both root and leaf tissue. A. marina roots may be employed as a biological indicator of environmental exposure of Cu, Pb and Zn and leaves for Zn, with temporal monitoring

Metal accumulation in the polychaete Hediste japonica with emphasis on interaction between heavy metals and petroleum hydrocarbons

International Nuclear Information System (INIS)

Sun Fuhong; Zhou Qixing

2007-01-01

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. - The influences of petroleum hydrocarbons on Cd and Cu accumulation in H. japonica depend on their concentration combinations and exposure time

Immunotoxicity of copper nanoparticle and copper sulfate in a common Indian earthworm.

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Gautam, Arunodaya; Ray, Abhishek; Mukherjee, Soumalya; Das, Santanu; Pal, Kunal; Das, Subhadeep; Karmakar, Parimal; Ray, Mitali; Ray, Sajal

2018-02-01

Copper oxide nanoparticles and copper sulfate are established contaminants of water and soil. Metaphire posthuma is a common variety of earthworm distributed in moist soil of Indian subcontinent. Comparative toxicity of copper nanoparticles and copper sulfate were investigated with reference to selected immune associated parameters of earthworm. Total count, phagocytic response, generation of cytotoxic molecules (superoxide anion, nitric oxide), activities of enzymes like phenoloxidase, superoxide dismutase, catalase, acid phosphatase, alkaline phosphatase and total protein of coelomocytes were estimated under the exposures of 100, 500, 1000mg of copper oxide nanoparticles and copper sulfate per kg of soil for 7 and 14 d. A significant decrease in the total coelomocyte count were recorded with maximum depletion as 15.45 ± 2.2 and 12.5 ± 2 × 10 4 cells/ml under the treatment of 1000mg/kg of copper nanoparticles and copper sulfate for 14 d respectively. A significant decrease in generation of nitric oxide and activity of phenoloxidase were recorded upon exposure of both toxins for 7 and 14 d indicating possible decline in cytotoxic status of the organism. A maximum inhibition of superoxide dismutase activity was recorded as 0.083 ± 0.0039 and 0.055 ± 0.0057 unit/mg protein/minute against 1000mg/kg of copper nanoparticles and copper sulfate treatment for 14 d respectively. Activities of catalase and alkaline phosphatase were inhibited by all experimental concentrations of both toxins in the coelomocytes of earthworm. These toxins were recorded to be modifiers of the major immune associated parameters of M. posthuma. Unrestricted contamination of soil by sulfate and oxide nanoparticles of copper may lead to an undesirable shift in the innate immunological status of earthworm leading to a condition of immune compromisation and shrinkage in population density of this species in its natural habitat. This article is the first time report of immunological toxicity of

Biliary copper excretion by hepatocyte lysosomes in the rat. Major excretory pathway in experimental copper overload

International Nuclear Information System (INIS)

Gross, J.B. Jr.; Myers, B.M.; Kost, L.J.; Kuntz, S.M.; LaRusso, N.F.

1989-01-01

We investigated the hypothesis that lysosomes are the main source of biliary copper in conditions of hepatic copper overload. We used a rat model of oral copper loading and studied the relationship between the biliary output of copper and lysosomal hydrolases. Male Sprague-Dawley rats were given tap water with or without 0.125% copper acetate for up to 36 wk. Copper loading produced a 23-fold increase in the hepatic copper concentration and a 30-65% increase in hepatic lysosomal enzyme activity. Acid phosphatase histochemistry showed that copper-loaded livers contained an increased number of hepatocyte lysosomes; increased copper concentration of these organelles was confirmed directly by both x ray microanalysis and tissue fractionation. The copper-loaded rats showed a 16-fold increase in biliary copper output and a 50-300% increase in biliary lysosomal enzyme output. In the basal state, excretory profiles over time were similar for biliary outputs of lysosomal enzymes and copper in the copper-loaded animals but not in controls. After pharmacologic stimulation of lysosomal exocytosis, biliary outputs of copper and lysosomal hydrolases in the copper-loaded animals remained coupled: injection of colchicine or vinblastine produced an acute rise in the biliary output of both lysosomal enzymes and copper to 150-250% of baseline rates. After these same drugs, control animals showed only the expected increase in lysosomal enzyme output without a corresponding increase in copper output. We conclude that the hepatocyte responds to an increased copper load by sequestering excess copper in an increased number of lysosomes that then empty their contents directly into bile. The results provide direct evidence that exocytosis of lysosomal contents into biliary canaliculi is the major mechanism for biliary copper excretion in hepatic copper overload

Underwater explosive compaction-sintering of tungsten-copper coating on a copper surface

Science.gov (United States)

Chen, Xiang; Li, Xiaojie; Yan, Honghao; Wang, Xiaohong; Chen, Saiwei

2018-01-01

This study investigated underwater explosive compaction-sintering for coating a high-density tungsten-copper composite on a copper surface. First, 50% W-50% Cu tungsten-copper composite powder was prepared by mechanical alloying. The composite powder was pre-compacted and sintered by hydrogen. Underwater explosive compaction was carried out. Finally, a high-density tungsten-copper coating was obtained by diffusion sintering of the specimen after explosive compaction. A simulation of the underwater explosive compaction process showed that the peak value of the pressure in the coating was between 3.0 and 4.8 GPa. The hardness values of the tungsten-copper layer and the copper substrate were in the range of 87-133 and 49 HV, respectively. The bonding strength between the coating and the substrate was approximately 100-105 MPa.

Mechanisms of redox interactions of bilirubin with copper and the effects of penicillamine.

Science.gov (United States)

Božić, Bojana; Korać, Jelena; Stanković, Dalibor M; Stanić, Marina; Popović-Bijelić, Ana; Bogdanović Pristov, Jelena; Spasojević, Ivan; Bajčetić, Milica

2017-12-25

Toxic effects of unconjugated bilirubin (BR) in neonatal hyperbilirubinemia have been related to redox and/or coordinate interactions with Cu 2+ . However, the development and mechanisms of such interactions at physiological pH have not been resolved. This study shows that BR reduces Cu 2+ to Cu 1+ in 1:1 stoichiometry. Apparently, BR undergoes degradation, i.e. BR and Cu 2+ do not form stable complexes. The binding of Cu 2+ to inorganic phosphates, liposomal phosphate groups, or to chelating drug penicillamine, impedes redox interactions with BR. Cu 1+ undergoes spontaneous oxidation by O 2 resulting in hydrogen peroxide accumulation and hydroxyl radical production. In relation to this, copper and BR induced synergistic oxidative/damaging effects on erythrocytes membrane, which were alleviated by penicillamine. The production of reactive oxygen species by BR and copper represents a plausible cause of BR toxic effects and cell damage in hyperbilirubinemia. Further examination of therapeutic potentials of copper chelators in the treatment of severe neonatal hyperbilirubinemia is needed. Copyright © 2017 Elsevier B.V. All rights reserved.

Microbial incidence on copper and titanium embedded in compacted bentonite clay

Energy Technology Data Exchange (ETDEWEB)

Persson, Joergen; Lydmark, Sara; Edlund, Johanna; Paeaejaervi, Anna; Pedersen, Karsten (Microbial Analytics Sweden AB (Sweden))

2011-10-15

The incidence of bacteria on metal surfaces was examined in an experimental setting simulating conditions of the proposed Swedish concept for disposal of spent nuclear fuel. Titanium and copper rods were embedded in compacted bentonite clay saturated with groundwater collected at a depth of 450 m. Bentonite blocks were exposed to an external flux of groundwater with or without added lactate or H{sub 2} for up to 203 days. Bacterial accumulation on metal rods and in the surrounding bentonite was analyzed using real-time quantitative PCR (qPCR), with genetic markers for overall bacterial presence (16S rDNA) as well as specific for sulfate-reducing bacteria (apsA). Clay species composition was analyzed by cloning and sequencing 16S rDNA extracted from the clay. Results suggest limited bacterial accumulation on metal surfaces, amounting to a maximum of approximately 106 apsA copies cm-2, corresponding to a 3.7% coverage of metal surfaces. Bacterial species composition appeared to be a mix of species originating from the bentonite clay and from the added groundwater, including an apparently high proportion of sulfate-reducing bacteria. While titanium surfaces exhibited higher bacterial presence than did copper surfaces, neither the degree of bentonite compaction nor the addition of lactate or H{sub 2} appeared to have any effect on the bacterial incidence on metal surfaces

Microbial incidence on copper and titanium embedded in compacted bentonite clay

International Nuclear Information System (INIS)

Persson, Joergen; Lydmark, Sara; Edlund, Johanna; Paeaejaervi, Anna; Pedersen, Karsten

2011-10-01

The incidence of bacteria on metal surfaces was examined in an experimental setting simulating conditions of the proposed Swedish concept for disposal of spent nuclear fuel. Titanium and copper rods were embedded in compacted bentonite clay saturated with groundwater collected at a depth of 450 m. Bentonite blocks were exposed to an external flux of groundwater with or without added lactate or H 2 for up to 203 days. Bacterial accumulation on metal rods and in the surrounding bentonite was analyzed using real-time quantitative PCR (qPCR), with genetic markers for overall bacterial presence (16S rDNA) as well as specific for sulfate-reducing bacteria (apsA). Clay species composition was analyzed by cloning and sequencing 16S rDNA extracted from the clay. Results suggest limited bacterial accumulation on metal surfaces, amounting to a maximum of approximately 10 6 apsA copies cm -2 , corresponding to a 3.7% coverage of metal surfaces. Bacterial species composition appeared to be a mix of species originating from the bentonite clay and from the added groundwater, including an apparently high proportion of sulfate-reducing bacteria. While titanium surfaces exhibited higher bacterial presence than did copper surfaces, neither the degree of bentonite compaction nor the addition of lactate or H 2 appeared to have any effect on the bacterial incidence on metal surfaces

Sulfidation treatment of copper-containing plating sludge towards copper resource recovery.

Science.gov (United States)

Kuchar, D; Fukuta, T; Onyango, M S; Matsuda, H

2006-11-02

The present study is concerned with the sulfidation treatment of copper-containing plating sludge towards copper resource recovery by flotation of copper sulfide from treated sludge. The sulfidation treatment was carried out by contacting simulated or real copper plating sludge with Na(2)S solution for a period of 5 min to 24 h. The initial molar ratio of S(2-) to Cu(2+) (S(2-) to Me(2+) in the case of real sludge) was adjusted to 1.00, 1.25 or 1.50, while the solid to liquid ratio was set at 1:50. As a result, it was found that copper compounds were converted to various copper sulfides within the first 5 min. In the case of simulated copper sludge, CuS was identified as the main sulfidation product at the molar ratio of S(2-) to Cu(2+) of 1.00, while Cu(7)S(4) (Roxbyite) was mainly found at the molar ratios of S(2-) to Cu(2+) of 1.50 and 1.25. Based on the measurements of oxidation-reduction potential, the formation of either CuS or Cu(7)S(4) at different S(2-) to Cu(2+) molar ratios was attributed to the changes in the oxidation-reduction potential. By contrast, in the case of sulfidation treatment of real copper sludge, CuS was predominantly formed, irrespective of S(2-) to Me(2+) molar ratio.

Accumulation of Heavy Metals in Crayfish and Fish from Selected Czech Reservoirs

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Iryna Kuklina

2014-01-01

Full Text Available To evaluate the accumulation of aluminium, cadmium, chromium, copper, lead, mercury, nickel, and zinc in crayfish and fish organ tissues, specimens from three drinking water reservoirs (Boskovice, Landštejn, and Nová Říše and one contaminated site (Darkovské moře in the Czech Republic were examined. Crayfish hepatopancreas was confirmed to be the primary accumulating site for the majority of metals (Cu > Zn > Ni > Cd > Cr, while Hg and Cr were concentrated in abdominal muscle, and Al and Pb were concentrated in gill. Metals found in Nová Říše specimens included Cu > Zn > Ni and those found in Boskovice included Zn > Hg > Cr. Cd concentrations were observed only in Landštejn specimens, while contaminated Darkovské moře specimens showed the highest levels of accumulation (Cu > Al > Zn > Pb. The majority of evaluated metals were found in higher concentrations in crayfish: Cu > Al > Zn > Ni > Cr > Cd > Pb, with Hg being the only metal accumulating higher in fish. Due to accumulation similarities of Al in crayfish and fish gill, differences of Hg in muscle, and features noted for the remaining metals in examined tissues, biomonitoring should incorporate both crayfish and fish to produce more relevant water quality surveys.

Aided Phytostabilization of Copper Contaminated Soils with L. Perenne and Mineral Sorbents as Soil Amendments

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Radziemska, Maja

2017-09-01

The present study was designed to assess phytostabilization strategies for the treatment of soil co-contaminated by increasing levels of copper with the application mineral amendments (chalcedonite, zeolite, dolomite). From the results it will be possible to further elucidate the benefits or potential risks derived from the application of different types of mineral amendments in the remediation of a copper contaminated soil. A glasshouse pot experiment was designed to evaluate the potential use of different amendments as immobilizing agents in the aided phytostabilization of Cu-contaminated soil using ryegrass (Lolium perenne L.). The content of trace elements in plants and total in soil, were determined using the method of spectrophotometry. All of the investigated element contents in the tested parts of L. perenne were significantly different in the case of applying mineral amendments to the soil, as well as increasing concentrations of copper. The greatest average above-ground biomass was observed for soil amended with chalcedonite. In this experiment, all analyzed metals accumulated predominantly in the roots of the tested plant. In general, applying mineral amendments to soil contributed to decreased levels of copper concentrations.

Ecophysiological tolerance of duckweeds exposed to copper

Energy Technology Data Exchange (ETDEWEB)

Kanoun-Boule, Myriam [Centre for Functional Ecology, Department of Botany, University of Coimbra, Coimbra 3000-456 (Portugal)], E-mail: mkb@ci.uc.pt; Vicente, Joaquim A.F.; Nabais, Cristina [Centre for Functional Ecology, Department of Botany, University of Coimbra, Coimbra 3000-456 (Portugal); Prasad, M.N.V. [Department of Plant Sciences, University of Hyderabad, Hyderabad 500046 (India); Freitas, Helena [Centre for Functional Ecology, Department of Botany, University of Coimbra, Coimbra 3000-456 (Portugal)

2009-01-18

Although essential for plants, copper can be toxic when present in supra-optimal concentrations. Metal polluted sites, due to their extreme conditions, can harbour tolerant species and/or ecotypes. In this work we aimed to compare the physiological responses to copper exposure and the uptake capacities of two species of duckweed, Lemna minor (Lm(EC1)) and Spirodela polyrrhiza (SP), from an abandoned uranium mine with an ecotype of L. minor (Lm(EC2)) from a non-contaminated pond. From the lowest Cu concentration exposure (25 {mu}M) to the highest (100 {mu}M), Lm(EC2) accumulated higher amounts of copper than Lm(EC1) and SP. Dose-response curves showed that Cu content accumulated by Lm(EC2) increases linearly with Cu treatment concentrations (r{sup 2} = 0.998) whereas quadratic models were more suitable for Lm(EC1) and SP (r{sup 2} = 0.999 and r{sup 2} = 0.998 for Lm(EC1) and SP, respectively). A significant concentration-dependent decline of chlorophyll a (chl a) and carotenoid occurred as a consequence of Cu exposure. These declines were significant for Lm(EC2) exposed to the lowest Cu concentration (25 {mu}M) whereas for Lm(EC1) and SP a significant decrease in chl a and carotenoids was observed only at 50 and 100 {mu}M-Cu. Electric conductivity (EC) and malondialdehyde (MDA) content increased after Cu exposure, indicating oxidative stress. Significant increase of EC was observed in Lm(EC2) for all Cu concentrations whereas the increase for Lm(EC1) and SP became significant only after an exposure to 50 {mu}M-Cu. On the contrary, for Lm(EC1), SP, and Lm(EC2), MDA content significantly increased even at the lowest concentration. Protein content and catalase (CAT) activity showed a decrease with an increase in Cu concentration. For the species Lm(EC1) and SP, a significant effect of copper on CAT activity was observed only at the highest concentration (100 {mu}M-Cu) whereas, for Lm(EC2), this effect started to be significant after an exposure to 50 {mu

Ecophysiological tolerance of duckweeds exposed to copper

International Nuclear Information System (INIS)

Kanoun-Boule, Myriam; Vicente, Joaquim A.F.; Nabais, Cristina; Prasad, M.N.V.; Freitas, Helena

2009-01-01

Although essential for plants, copper can be toxic when present in supra-optimal concentrations. Metal polluted sites, due to their extreme conditions, can harbour tolerant species and/or ecotypes. In this work we aimed to compare the physiological responses to copper exposure and the uptake capacities of two species of duckweed, Lemna minor (Lm(EC1)) and Spirodela polyrrhiza (SP), from an abandoned uranium mine with an ecotype of L. minor (Lm(EC2)) from a non-contaminated pond. From the lowest Cu concentration exposure (25 μM) to the highest (100 μM), Lm(EC2) accumulated higher amounts of copper than Lm(EC1) and SP. Dose-response curves showed that Cu content accumulated by Lm(EC2) increases linearly with Cu treatment concentrations (r 2 = 0.998) whereas quadratic models were more suitable for Lm(EC1) and SP (r 2 = 0.999 and r 2 = 0.998 for Lm(EC1) and SP, respectively). A significant concentration-dependent decline of chlorophyll a (chl a) and carotenoid occurred as a consequence of Cu exposure. These declines were significant for Lm(EC2) exposed to the lowest Cu concentration (25 μM) whereas for Lm(EC1) and SP a significant decrease in chl a and carotenoids was observed only at 50 and 100 μM-Cu. Electric conductivity (EC) and malondialdehyde (MDA) content increased after Cu exposure, indicating oxidative stress. Significant increase of EC was observed in Lm(EC2) for all Cu concentrations whereas the increase for Lm(EC1) and SP became significant only after an exposure to 50 μM-Cu. On the contrary, for Lm(EC1), SP, and Lm(EC2), MDA content significantly increased even at the lowest concentration. Protein content and catalase (CAT) activity showed a decrease with an increase in Cu concentration. For the species Lm(EC1) and SP, a significant effect of copper on CAT activity was observed only at the highest concentration (100 μM-Cu) whereas, for Lm(EC2), this effect started to be significant after an exposure to 50 μM-Cu. Superoxide dismutase (SOD) activity

Critical review: Copper runoff from outdoor copper surfaces at atmospheric conditions.

Science.gov (United States)

Hedberg, Yolanda S; Hedberg, Jonas F; Herting, Gunilla; Goidanich, Sara; Odnevall Wallinder, Inger

2014-01-01

This review on copper runoff dispersed from unsheltered naturally patinated copper used for roofing and facades summarizes and discusses influencing factors, available literature, and predictive models, and the importance of fate and speciation for environmental risk assessment. Copper runoff from outdoor surfaces is predominantly governed by electrochemical and chemical reactions and is highly dependent on given exposure conditions (size, inclination, geometry, degree of sheltering, and orientation), surface parameters (age, patina composition, and thickness), and site-specific environmental conditions (gaseous pollutants, chlorides, rainfall characteristics (amount, intensity, pH), wind direction, temperature, time of wetness, season). The corrosion rate cannot be used to assess the runoff rate. The extent of released copper varies largely between different rain events and is related to dry and wet periods, dry deposition prior to the rain event and prevailing rain and patina characteristics. Interpretation and use of copper runoff data for environmental risk assessment and management need therefore to consider site-specific factors and focus on average data of long-term studies (several years). Risk assessments require furthermore that changes in copper speciation, bioavailability aspects, and potential irreversible retention on solid surfaces are considered, factors that determine the environmental fate of copper runoff from outdoor surfaces.

Intracellular targeting of annexin A2 inhibits tumor cell adhesion, migration, and in vivo grafting.

Science.gov (United States)

Staquicini, Daniela I; Rangel, Roberto; Guzman-Rojas, Liliana; Staquicini, Fernanda I; Dobroff, Andrey S; Tarleton, Christy A; Ozbun, Michelle A; Kolonin, Mikhail G; Gelovani, Juri G; Marchiò, Serena; Sidman, Richard L; Hajjar, Katherine A; Arap, Wadih; Pasqualini, Renata

2017-06-26

Cytoskeletal-associated proteins play an active role in coordinating the adhesion and migration machinery in cancer progression. To identify functional protein networks and potential inhibitors, we screened an internalizing phage (iPhage) display library in tumor cells, and selected LGRFYAASG as a cytosol-targeting peptide. By affinity purification and mass spectrometry, intracellular annexin A2 was identified as the corresponding binding protein. Consistently, annexin A2 and a cell-internalizing, penetratin-fused version of the selected peptide (LGRFYAASG-pen) co-localized and specifically accumulated in the cytoplasm at the cell edges and cell-cell contacts. Functionally, tumor cells incubated with LGRFYAASG-pen showed disruption of filamentous actin, focal adhesions and caveolae-mediated membrane trafficking, resulting in impaired cell adhesion and migration in vitro. These effects were paralleled by a decrease in the phosphorylation of both focal adhesion kinase (Fak) and protein kinase B (Akt). Likewise, tumor cells pretreated with LGRFYAASG-pen exhibited an impaired capacity to colonize the lungs in vivo in several mouse models. Together, our findings demonstrate an unrecognized functional link between intracellular annexin A2 and tumor cell adhesion, migration and in vivo grafting. Moreover, this work uncovers a new peptide motif that binds to and inhibits intracellular annexin A2 as a candidate therapeutic lead for potential translation into clinical applications.